http://205.166.159.208/wiki/api.php?action=feedcontributions&feedformat=atom&user=Ssaey 2026-05-03T15:38:16Z User contributions MediaWiki 1.35.5 http://205.166.159.208/wiki/index.php?title=Stephanie_Saey&diff=8909 2018-05-11T01:23:15Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Personal Information== --&gt;<br /> &lt;!-- ==Undergraduate Research Activities== --&gt;<br /> &lt;!-- ==Presentations== --&gt;<br /> &lt;!-- ==Publications== --&gt;<br /> &lt;!-- ==Activities== --&gt;<br /> &lt;!-- ==Career Goals== --&gt;<br /> &lt;!-- --&gt;<br /> &lt;!-- --&gt;<br /> You have reached the personal page of Stephanie Saey. <br /> <br /> ==Personal Information==<br /> Graduated (Spring 2018) Biochemistry and Biopsychology Double Major <br /> <br /> Hometown: Galesburg, Illinois<br /> <br /> ==Undergraduate Research Activities==<br /> <br /> Summer 2015: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Kristin Larson <br /> *Reviewed literature surrounding the psychological phenomenon of the &quot;third place&quot;<br /> *Developed a research proposal to study the relationship between geographical setting of the &quot;third place&quot; coffee shop and individual place attachment<br /> *Wrote HSRB and developed survey<br /> *Collected data in Chicago, Illinois, and Galesburg, Illinois<br /> *Analyzed data using Statistical Software Minitab<br /> <br /> Summer 2016: Freeman Grant Recipient, Travelled to Singapore for research with Dr. Kristin Larson <br /> *Collaborated with Dr. Kristin Larson and two undergraduate colleagues to develop a research proposal to study in Singapore <br /> *Submitted the research proposal to the ASIANetwork Student-Faculty Fellows Program <br /> *Received the ASIANetwork grant, funded by the Freeman Foundation, to study for three weeks in Singapore <br /> *Wrote HSRB and developed survey to study the relationship between &quot;third places&quot; and perceived stress levels <br /> *Currently (Fall 2016 semester) in the process of analyzing data and writing a post-report <br /> <br /> Summer 2016: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Bradley Sturgeon and Dr. Laura Moore <br /> *Served as a student mentor for incoming freshmen participating in the three-week program<br /> *Led discussions and small experiments on the chemistry of basic baking (i.e., gluten formation and yeast activity)<br /> *Performed an extraction of gluten proteins from various types of flour dough and analyzed the extractions through SDS-PAGE <br /> *Mentored the students in developing a presentation for the SOfIA colloquium <br /> *Helped supervise the making of final posters for the project <br /> <br /> Fall 2016-Spring 2017: Chem 430 with B. Sturgeon, worked as a mentee of Nadia Ayala<br /> Spring 2017: Continued research with the secondary plant metabolite, curcumin:<br /> [[Curcumin_Research|Curcumin Research]]<br /> Fall 2017-Spring 2018: Worked to oxidize lignin monomers (mentee: Zelinda Taylor): <br /> [[Oxidation_of_Lignin_Monomers|Oxidation of Lignin Monomers]]<br /> ==Presentations==<br /> <br /> August 2015: '''SOfIA Presentation'''<br /> <br /> Poster Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> Spring 2016: '''ILLOWA Undergraduate Research Conference''' Macomb, IL<br /> <br /> Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> August 2016: '''SOfIA Presentation'''<br /> <br /> Poster Titles: ''The Chemistry of Baking: The Basics, The Chemistry of Baking: Experiments''<br /> <br /> <br /> ==Publications==<br /> Spring 2016: '''The 2016-17 Midwest Journal of Undergraduate Research'''<br /> <br /> Publication Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> ==Activities==<br /> <br /> <br /> ==Career Goals==<br /> <br /> Psychiatry</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8908 2018-05-11T01:22:17Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stephanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A). Coniferyl alcohol was oxidized in a beaker under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. A standard solution of the coniferyl alcohol monomer was made by dissolving 0.0385g of the product in 100 mL of a 50/50 dioxane/pH5 buffer. The first reaction was carried out with 5 mL of the stock solution (final concentration of 2mM coniferyl alcohol) in the presence of 10 microliters of HRP catalyst. The second reaction was carried out with 5 mL coniferyl alcohol in the presence of both 10 microliters HRP and 5 microliters of a 0.5M hydrogen peroxide solution (0.5 mM final concentration hydrogen peroxide). Subsequently, a third reaction was carried out involving 5 mL of coniferyl alcohol, 10 microliters of catalase, and 5 microliters of HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|left|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===HPLC Analysis===<br /> Each reaction, along with the standard coniferyl alcohol solution, was analyzed using HPLC with an ACN/H20 gradient run for 35 minutes. The first 10 minutes were ran at 100% H20, followed by 80%ACN/20%H20 for minutes 10-30, and finishing from minutes 30-35 at 100% H20 (Figure 2). The exact method is saved in the HLPC instrument as &quot;pcou_100417_35_SAZT.&quot; <br /> [[File:Hplc data.png|400px|thumb|center|Figure 2: HPLC data of HRP/H2O2 oxidation of coniferyl alcohol using 50/50 dioxane/pH 5 buffer.]]<br /> <br /> ===Dicussion===<br /> This work is still in progress. Zelinda Taylor will be continuing the project as a Doc Kieft Scholar during the summer of 2018. <br /> <br /> ===Poster===<br /> The following poster was presented by Stephanie Saey at the National ACS Meeting in New Orleans, Louisiana during the Spring of 2018:<br /> [[File:Poster for ACS 2018.png|400px|thumb|center|]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8907 2018-05-11T01:21:54Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A). Coniferyl alcohol was oxidized in a beaker under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. A standard solution of the coniferyl alcohol monomer was made by dissolving 0.0385g of the product in 100 mL of a 50/50 dioxane/pH5 buffer. The first reaction was carried out with 5 mL of the stock solution (final concentration of 2mM coniferyl alcohol) in the presence of 10 microliters of HRP catalyst. The second reaction was carried out with 5 mL coniferyl alcohol in the presence of both 10 microliters HRP and 5 microliters of a 0.5M hydrogen peroxide solution (0.5 mM final concentration hydrogen peroxide). Subsequently, a third reaction was carried out involving 5 mL of coniferyl alcohol, 10 microliters of catalase, and 5 microliters of HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|left|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===HPLC Analysis===<br /> Each reaction, along with the standard coniferyl alcohol solution, was analyzed using HPLC with an ACN/H20 gradient run for 35 minutes. The first 10 minutes were ran at 100% H20, followed by 80%ACN/20%H20 for minutes 10-30, and finishing from minutes 30-35 at 100% H20 (Figure 2). The exact method is saved in the HLPC instrument as &quot;pcou_100417_35_SAZT.&quot; <br /> [[File:Hplc data.png|400px|thumb|center|Figure 2: HPLC data of HRP/H2O2 oxidation of coniferyl alcohol using 50/50 dioxane/pH 5 buffer.]]<br /> <br /> ===Dicussion===<br /> This work is still in progress. Zelinda Taylor will be continuing the project as a Doc Kieft Scholar during the summer of 2018. <br /> <br /> ===Poster===<br /> The following poster was presented by Stephanie Saey at the National ACS Meeting in New Orleans, Louisiana during the Spring of 2018:<br /> [[File:Poster for ACS 2018.png|400px|thumb|center|]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8906 2018-05-11T01:21:27Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A). Coniferyl alcohol was oxidized in a beaker under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. A standard solution of the coniferyl alcohol monomer was made by dissolving 0.0385g of the product in 100 mL of a 50/50 dioxane/pH5 buffer. The first reaction was carried out with 5 mL of the stock solution (final concentration of 2mM coniferyl alcohol) in the presence of 10 microliters of HRP catalyst. The second reaction was carried out with 5 mL coniferyl alcohol in the presence of both 10 microliters HRP and 5 microliters of a 0.5M hydrogen peroxide solution (0.5 mM final concentration hydrogen peroxide). Subsequently, a third reaction was carried out involving 5 mL of coniferyl alcohol, 10 microliters of catalase, and 5 microliters of HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|left|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===HPLC Analysis===<br /> Each reaction, along with the standard coniferyl alcohol solution, was analyzed using HPLC with an ACN/H20 gradient run for 35 minutes. The first 10 minutes were ran at 100% H20, followed by 80%ACN/20%H20 for minutes 10-30, and finishing from minutes 30-35 at 100% H20. The exact method is saved in the HLPC instrument as &quot;pcou_100417_35_SAZT.&quot; <br /> [[File:Hplc data.png|400px|thumb|center|Figure 2: HPLC data of HRP/H2O2 oxidation of coniferyl alcohol using 50/50 dioxane/pH 5 buffer.]]<br /> <br /> ===Dicussion===<br /> This work is still in progress. Zelinda Taylor will be continuing the project as a Doc Kieft Scholar during the summer of 2018. <br /> <br /> ===Poster===<br /> The following poster was presented by Stephanie Saey at the National ACS Meeting in New Orleans, Louisiana during the Spring of 2018:<br /> [[File:Poster for ACS 2018.png|400px|thumb|center|]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8905 2018-05-11T01:20:25Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|left|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A). Coniferyl alcohol was oxidized in a beaker under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. A standard solution of the coniferyl alcohol monomer was made by dissolving 0.0385g of the product in 100 mL of a 50/50 dioxane/pH5 buffer. The first reaction was carried out with 5 mL of the stock solution (final concentration of 2mM coniferyl alcohol) in the presence of 10 microliters of HRP catalyst. The second reaction was carried out with 5 mL coniferyl alcohol in the presence of both 10 microliters HRP and 5 microliters of a 0.5M hydrogen peroxide solution (0.5 mM final concentration hydrogen peroxide). Subsequently, a third reaction was carried out involving 5 mL of coniferyl alcohol, 10 microliters of catalase, and 5 microliters of HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|left|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===HPLC Analysis===<br /> Each reaction, along with the standard coniferyl alcohol solution, was analyzed using HPLC with an ACN/H20 gradient run for 35 minutes. The first 10 minutes were ran at 100% H20, followed by 80%ACN/20%H20 for minutes 10-30, and finishing from minutes 30-35 at 100% H20. The exact method is saved in the HLPC instrument as &quot;pcou_100417_35_SAZT.&quot; <br /> [[File:Hplc data.png|400px|thumb|left|Figure 2: HPLC data of HRP/H2O2 oxidation of coniferyl alcohol using 50/50 dioxane/pH 5 buffer.]]<br /> <br /> ===Dicussion===<br /> This work is still in progress. Zelinda Taylor will be continuing the project as a Doc Kieft Scholar during the summer of 2018. <br /> <br /> ===Poster===<br /> The following poster was presented by Stephanie Saey at the National ACS Meeting in New Orleans, Louisiana during the Spring of 2018:<br /> [[File:Poster for ACS 2018.png|400px|thumb|left]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8904 2018-05-11T01:19:37Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A). Coniferyl alcohol was oxidized in a beaker under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. A standard solution of the coniferyl alcohol monomer was made by dissolving 0.0385g of the product in 100 mL of a 50/50 dioxane/pH5 buffer. The first reaction was carried out with 5 mL of the stock solution (final concentration of 2mM coniferyl alcohol) in the presence of 10 microliters of HRP catalyst. The second reaction was carried out with 5 mL coniferyl alcohol in the presence of both 10 microliters HRP and 5 microliters of a 0.5M hydrogen peroxide solution (0.5 mM final concentration hydrogen peroxide). Subsequently, a third reaction was carried out involving 5 mL of coniferyl alcohol, 10 microliters of catalase, and 5 microliters of HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|none|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===HPLC Analysis===<br /> Each reaction, along with the standard coniferyl alcohol solution, was analyzed using HPLC with an ACN/H20 gradient run for 35 minutes. The first 10 minutes were ran at 100% H20, followed by 80%ACN/20%H20 for minutes 10-30, and finishing from minutes 30-35 at 100% H20. The exact method is saved in the HLPC instrument as &quot;pcou_100417_35_SAZT.&quot; <br /> [[File:Hplc data.png|400px|thumb|none|Figure 2: HPLC data of HRP/H2O2 oxidation of coniferyl alcohol using 50/50 dioxane/pH 5 buffer.]]<br /> <br /> ===Dicussion===<br /> This work is still in progress. Zelinda Taylor will be continuing the project as a Doc Kieft Scholar during the summer of 2018. <br /> <br /> ===Poster===<br /> The following poster was presented by Stephanie Saey at the National ACS Meeting in New Orleans, Louisiana during the Spring of 2018:<br /> [[File:Poster for ACS 2018.png|400px]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:Poster_for_ACS_2018.png&diff=8903 2018-05-11T01:19:22Z

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Ssaey http://205.166.159.208/wiki/index.php?title=File:Hplc_data.png&diff=8902 2018-05-11T01:15:25Z

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Ssaey http://205.166.159.208/wiki/index.php?title=File:Coniferyl.jpg&diff=8901 2018-05-11T00:59:25Z

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Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8900 2018-05-11T00:58:32Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP (Figure 1). Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|Figure 1: ESR data from HRP oxidation of lignin monomers]]<br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A and Image B). Coniferyl alcohol was oxidized under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. The first reaction was carried out with 2mM coniferyl alcohol in the presence of HRP catalyst. The second reaction was carried out with 2mM coniferyl alcohol in the presence of both HRP and 0.5 mM hydrogen peroxide. Subsequently, a third reaction was carried out involving 2mM coniferyl alcohol, catalase, and HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|none|Image A: Resonance structures of the coniferyl alcohol radical following oxidation. Asteriks indicate the radicals with the greatest propensity to undergo coupling reactions, as predicted by Heitner, Dimmel, and Schmidt (2010).]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8899 2018-05-11T00:55:25Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.<br /> <br /> ===Preliminary ESR data===<br /> Lignin monomer radicals were detected using 50/50 water/dioxane (pH 5) to solublize polymer products; immobilized HRP. Quantum calculations to analyze each spectra are in progress; however, for now it can most definitely be concluded that each monomer has the ability to form radicals under oxidative conditions. <br /> <br /> ===Oxidation of Coniferyl Alcohol===<br /> The chemical structure of coniferyl alcohol allows for various places where coupling may occur once the radical forms (Image A and Image B). Coniferyl alcohol was oxidized under various reaction conditions to gain insight into the number of products formed by these radical-radical coupling reactions. The first reaction was carried out with 2mM coniferyl alcohol in the presence of HRP catalyst. The second reaction was carried out with 2mM coniferyl alcohol in the presence of both HRP and 0.5 mM hydrogen peroxide. Subsequently, a third reaction was carried out involving 2mM coniferyl alcohol, catalase, and HRP. In the latter reaction, catalase was added last to the reaction beaker. <br /> <br /> [[File:coniferylpredictions.png|400px|thumb|none|Image A: Con]]<br /> <br /> ===References===<br /> Heitner, C., Dimmel, D.R., and Schmidt, J. (2010). Lignin and lignans: advances in chemistry. <br /> Li X, Yang Z, Zhao M. (2012). Neuroprotective effects of Flax Lignan against NMDA‐induced neurotoxicity in vitro. CNS Neuroscience &amp; Therapeutics, 18(11).927-933. <br /> Vanholm, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. Lignin biosynthesis and structure. (2010). Plant Physiology. doi:10.1104/pp.110.155119</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:Coniferylpredictions.png&diff=8898 2018-05-11T00:46:16Z

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Ssaey http://205.166.159.208/wiki/index.php?title=Oxidation_of_Lignin_Monomers&diff=8897 2018-05-11T00:34:55Z

<p>Ssaey: Created page with &quot;&lt;!-- ==Curcumin Research Initiative== --&gt; You have reached the page dedicated to the research of lignin monomers. This page was created by Stephanie Saey and...&quot;</p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of lignin monomers. This page was created by [[Stehanie_Saey|Stephanie Saey]] and will be maintained by Zelinda Taylor. Stephanie was a 2018 Biochemistry/Biopsychology graduate and Zelinda is currently a Junior Biochemistry research student.<br /> <br /> ===Abstract ===<br /> The biosynthesis of the lignin polymer occurs through oxidative coupling between three basic monomers: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol. The chemical structure of each lignin monomer includes a characteristic phenol group that contributes stability to the assumed radical intermediate formed upon oxidation. In this work, immobilized horseradish peroxidase was used to oxidize the coniferyl alcohol and to directly detect the radical intermediate using immobilized enzyme - ESR spectroscopy (IE-ESR). Oxidation products were analyzed by HPLC and findings will be discussed in terms of both lignin and lignan synthesis.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8896 2018-05-11T00:32:41Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> [[Stephanie_Saey|Stephanie Saey]] and Zelinda Taylor worked to oxidize the three lignin monomers in the Fall 2017/Spring 2018. An updated report of the research can be viewed here: [[Oxidation_of_Lignin_Monomers| Oxidation of Lignin Monomers]]<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by [[Stephanie_Saey|Stephanie Saey]]<br /> Please visit the following link for an updated report: [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8895 2018-05-11T00:32:14Z

<p>Ssaey: /* Lignans (General) */</p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> [[Stephanie_Saey|Stephanie Saey]] and Zelinda Taylor worked to oxidize the three lignin monomers in the Fall 2017/Spring 2018. An updated report of the research can be viewed here: [[Oxidation_of_Lignin_Monomers: Oxidation of Lignin Monomers]]<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by [[Stephanie_Saey|Stephanie Saey]]<br /> Please visit the following link for an updated report: [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8894 2018-05-11T00:27:54Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 80 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 300 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtOAc, 150 mL water, and 100 mL brine. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. The entire process resulted in approximately 250 mL of crude curcumin mixed with the solvents used for extraction.<br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Image C. Final Rf values were recorded. <br /> [[File:Example.jpg|200px|thumb|none|Image C: TLC results yielded three distinct and visible lines for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.]]<br /> <br /> :'''Concentration of Crude Extract'''<br /> ::Used rotovap (in Organic Chemistry lab) to concentrate the curcumin mixture before running Flash Chromatography.<br /> ::The cooler was set to 8.1 degrees Celcius and the water bath was set to 45 degrees Celcius. The speed was set at 5.<br /> ::Approximately 80 mL of the crude curcumin mixture was pipetted into a 500 mL rbf. The solvent was removed via the rotovap instrument for about 80 minutes. The resulting concentrated crude extract was transferred to a 20 mL glass vile and allowed to separate into a bottom, red layer (containing the curcuminoids) and a top, yellow layer (containing the oleoresin). The oleoresin was removed from the curcuminoids via a pipette and discarded. <br /> [[File:rotovapped crude mixture.png|200px|thumb|none|Image C: Crude extract following rotovapping; bottom layer (dark red) contains curcuminoids, top layer (yellow) is oleoresin]]<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks. The fractions for each peak were collected and stored in separate flasks. <br /> Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument.<br /> [[File:flashdata.png|500px|thumb|none|Figure 1: Flash Chromatography separation of the crude curcumin extract. The first peak was presumed to be bidemethoxycurcumin (curcumin III), the most polar of the three curcuminoids, followed by demethoxycurcumin (curcumin II) and curcumin I (most nonpolar).]] <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|Figure 2: HPLC chromatogram]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 45 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::Each fraction was rotovapped so that all solvent was removed, and then About 1mg of each curcumin product was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. Only fraction 1 (presumed to be bisdemethoxycurcumin) was analyzed at the time. <br /> [[File:fraction 1.png|500px|thumb|none|Figure 3: NMR spectrum for the first fraction collected during Flash Chromatography. Notable peaks include those in the aromatic region, as well as the methoxy region. The large peak at 4.5 is DMSO.]]<br /> <br /> ===Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcuminoids through NMR have led to further questions on the structures present within each curcumin sample. The first fraction eluted through Flash Chromatography using the method described above should be bisdemthoxycurcumin, as this derivative is the most polar. However, the literature agrees that the most nonpolar derivative (curcumin I) is the most abundant within the turmeric plant and therefore it remains unclear why the first peak yields the greatest absorbance and product if it is not curcumin I. It is possible that the first large peak on the Flash Chromatogram exists as a complex of all three derivates, and that the second and third peaks are unknown compounds such as the turmeric oleoresins. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction. The general consensus seems to be that the three derivatives are simply too similar in structure to be separated with the proposed method. <br /> <br /> ===Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8893 2018-05-11T00:18:55Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===2. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===3. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 80 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 300 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtOAc, 150 mL water, and 100 mL brine. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. The entire process resulted in approximately 250 mL of crude curcumin mixed with the solvents used for extraction.<br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Image C. Final Rf values were recorded. <br /> [[File:Example.jpg|200px|thumb|none|Image C: TLC results yielded three distinct and visible lines for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.]]<br /> <br /> :'''Concentration of Crude Extract'''<br /> ::Used rotovap (in Organic Chemistry lab) to concentrate the curcumin mixture before running Flash Chromatography.<br /> ::The cooler was set to 8.1 degrees Celcius and the water bath was set to 45 degrees Celcius. The speed was set at 5.<br /> ::Approximately 80 mL of the crude curcumin mixture was pipetted into a 500 mL rbf. The solvent was removed via the rotovap instrument for about 80 minutes. The resulting concentrated crude extract was transferred to a 20 mL glass vile and allowed to separate into a bottom, red layer (containing the curcuminoids) and a top, yellow layer (containing the oleoresin). The oleoresin was removed from the curcuminoids via a pipette and discarded. <br /> [[File:rotovapped crude mixture.png|200px|thumb|none|Image C: Crude extract following rotovapping; bottom layer (dark red) contains curcuminoids, top layer (yellow) is oleoresin]]<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks. The fractions for each peak were collected and stored in separate flasks. <br /> Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument.<br /> [[File:flashdata.png|500px|thumb|none|Figure 1: Flash Chromatography separation of the crude curcumin extract. The first peak was presumed to be bidemethoxycurcumin (curcumin III), the most polar of the three curcuminoids, followed by demethoxycurcumin (curcumin II) and curcumin I (most nonpolar).]] <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|Figure 2: HPLC chromatogram]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 45 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::Each fraction was rotovapped so that all solvent was removed, and then About 1mg of each curcumin product was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. Only fraction 1 (presumed to be bisdemethoxycurcumin) was analyzed at the time. <br /> [[File:fraction 1.png|400px|thmb|none|Figure 3: NMR spectrum for the first fraction collected during Flash Chromatography. Notable peaks include those in the aromatic region, as well as the methoxy region. The large peak at 4.5 is DMSO.]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:Fraction_1.png&diff=8892 2018-05-11T00:13:55Z

<p>Ssaey: File uploaded with MsUpload</p> <hr /> <div>File uploaded with MsUpload</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8891 2018-05-11T00:09:37Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===2. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===3. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 80 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 300 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtOAc, 150 mL water, and 100 mL brine. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. The entire process resulted in approximately 250 mL of crude curcumin mixed with the solvents used for extraction.<br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC results yielded three distinct and visible lines for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.]]<br /> <br /> :'''Concentration of Crude Extract'''<br /> ::Used rotovap (in Organic Chemistry lab) to concentrate the curcumin mixture before running Flash Chromatography.<br /> ::The cooler was set to 8.1 degrees Celcius and the water bath was set to 45 degrees Celcius. The speed was set at 5.<br /> ::Approximately 80 mL of the crude curcumin mixture was pipetted into a 500 mL rbf. The solvent was removed via the rotovap instrument for about 80 minutes. The resulting concentrated crude extract was transferred to a 20 mL glass vile and allowed to separate into a bottom, red layer (containing the curcuminoids) and a top, yellow layer (containing the oleoresin). The oleoresin was removed from the curcuminoids via a pipette and discarded. <br /> [[File:rotovapped crude mixture.png|200px|Image C: Crude extract following rotovapping; bottom layer (dark red) contains curcuminoids, top layer (yellow) is oleoresin]]<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks.** The fractions for each peak were collected and stored in separate flasks. <br /> **Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 45 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks.** The fractions for each peak were collected and stored in separate flasks. <br /> *Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument. <br /> [[File:flashdata.png|500px|Flash Chromatography separation of the crude curcumin extract. The first peak was presumed to be bidemethoxycurcumin (curcumin III), the most polar of the three curcuminoids, followed by demethoxycurcumin (curcumin II) and curcumin I (most nonpolar).]]<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::Each fraction was rotovapped so that all solvent was removed, and then About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8888 2018-05-11T00:06:48Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===2. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===3. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 80 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 300 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtOAc, 150 mL water, and 100 mL brine. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. The entire process resulted in approximately 250 mL of crude curcumin mixed with the solvents used for extraction.<br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Concentration of Crude Extract'''<br /> ::Used rotovap (in Organic Chemistry lab) to concentrate the curcumin mixture before running Flash Chromatography.<br /> ::The cooler was set to 8.1 degrees Celcius and the water bath was set to 45 degrees Celcius. The speed was set at 5.<br /> ::Approximately 80 mL of the crude curcumin mixture was pipetted into a 500 mL rbf. The solvent was removed via the rotovap instrument for about 80 minutes. The resulting concentrated crude extract was transferred to a 20 mL glass vile and allowed to separate into a bottom, red layer (containing the curcuminoids) and a top, yellow layer (containing the oleoresin). The oleoresin was removed from the curcuminoids via a pipette and discarded. <br /> [[File:rotovapped crude mixture.png|200px|Image C: Crude extract following rotovapping; bottom layer (dark red) contains curcuminoids, top layer (yellow) is oleoresin]]<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks.** The fractions for each peak were collected and stored in separate flasks. <br /> **Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 45 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks.** The fractions for each peak were collected and stored in separate flasks. <br /> *Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument. <br /> [[File:flashdata.png|500px|Flash Chromatography separation of the crude curcumin extract. The first peak was presumed to be bidemethoxycurcumin (curcumin III), the most polar of the three curcuminoids, followed by demethoxycurcumin (curcumin II) and curcumin I (most nonpolar).]]<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::Each fraction was rotovapped so that all solvent was removed, and then About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8886 2018-05-10T23:57:05Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===2. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===3. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 80 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 300 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtOAc, 150 mL water, and 100 mL brine. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. The entire process resulted in approximately 250 mL of crude curcumin mixed with the solvents used for extraction.<br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Concentration of Crude Extract'''<br /> ::Used rotovap (in Organic Chemistry lab) to concentrate the curcumin mixture before running Flash Chromatography.<br /> ::The cooler was set to 8.1 degrees Celcius and the water bath was set to 45 degrees Celcius. The speed was set at 5.<br /> ::Approximately 80 mL of the crude curcumin mixture was pipetted into a 500 mL rbf. The solvent was removed via the rotovap instrument for about 80 minutes. The resulting concentrated crude extract was transferred to a 20 mL glass vile and allowed to separate into a bottom, red layer (containing the curcuminoids) and a top, yellow layer (containing the oleoresin). The oleoresin was removed from the curcuminoids via a pipette and discarded. <br /> [[File:rotovapped crude mixture.png|400px]]<br /> <br /> :'''Flash Column Chromatography:'''<br /> ::An 100g gold C18 reverse phase Column was used to run 2mL of the concentrated curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 90 min (start: 10% ACN/100% H2O, finish: 100% ACN) that yielded three distinct peaks.** The fractions for each peak were collected and stored in separate flasks. <br /> **Method is saved as &quot;ssaeycurcuminsummer17&quot; in the Flash Chromatography instrument. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 45 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.[[File:flashdata.png|400px]]<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:Flashdata.png&diff=8885 2018-05-10T23:56:48Z

<p>Ssaey: File uploaded with MsUpload</p> <hr /> <div>File uploaded with MsUpload</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:Rotovapped_crude_mixture.png&diff=8876 2018-05-10T23:48:25Z

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Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8865 2018-05-10T23:29:32Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> Structures retrieved from: [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8862 2018-05-10T23:27:54Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> :[[Media:Turmeric ref 01.pdf|Reference]]<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8861 2018-05-10T23:27:02Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ''Procedure for extraction was modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8859 2018-05-10T23:25:03Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ===Flash Chromatography Data=== <br /> <br /> We used a 15.5mL C18 Column to run 1mL of our Curcumin extract. This column uses acetyl nitrile and water to run the liquid phase through the column. We ran a gradient for 25 min and got three distinct peaks which we then proceeded to run an HPLC method on.<br /> <br /> ===Sourcing turmeric===<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ===Overview of Proposed Methodology===<br /> ''Procedure is modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> <br /> *Extract curcuminoids with MeOH under soxhlet extraction.<br /> *Partition MeOH supernatant with ethyl acetate (AcOEt) and water for purification purposes.<br /> *Fractionate AcOEt extract by silica gel column chromatography to yield AcOEt curcumin eluents. <br /> *Fractionate AcOEt eluents by silica gel TLC using chloroform: acetone = 8:1 as the solvent system. <br /> *Purify each eluent by removing organic solvent (?) and analyzing under HPLC.<br /> <br /> ===Extracting Curcuminoids under Soxhlet Distillation===<br /> *Weigh out 80.00g of turmeric powder with an analytical balance. <br /> *Measure out 300.00mL MeOH using a burette. <br /> *Place above compound and solvent in stillpot of soxhlet apparatus. <br /> *Carefully turn on heat source and water source. <br /> *Run extraction for 6-7 hours.<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8858 2018-05-10T23:23:03Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> ===Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> &lt;!-- ==The Three Curcuminoids== --&gt;<br /> <br /> <br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> <br /> <br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ===Flash Chromatography Data=== <br /> <br /> We used a 15.5mL C18 Column to run 1mL of our Curcumin extract. This column uses acetyl nitrile and water to run the liquid phase through the column. We ran a gradient for 25 min and got three distinct peaks which we then proceeded to run an HPLC method on.<br /> <br /> ===Sourcing turmeric===<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ===Overview of Proposed Methodology===<br /> ''Procedure is modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> <br /> *Extract curcuminoids with MeOH under soxhlet extraction.<br /> *Partition MeOH supernatant with ethyl acetate (AcOEt) and water for purification purposes.<br /> *Fractionate AcOEt extract by silica gel column chromatography to yield AcOEt curcumin eluents. <br /> *Fractionate AcOEt eluents by silica gel TLC using chloroform: acetone = 8:1 as the solvent system. <br /> *Purify each eluent by removing organic solvent (?) and analyzing under HPLC.<br /> <br /> ===Extracting Curcuminoids under Soxhlet Distillation===<br /> *Weigh out 80.00g of turmeric powder with an analytical balance. <br /> *Measure out 300.00mL MeOH using a burette. <br /> *Place above compound and solvent in stillpot of soxhlet apparatus. <br /> *Carefully turn on heat source and water source. <br /> *Run extraction for 6-7 hours.<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8857 2018-05-10T23:22:48Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> ===2. Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> &lt;!-- ==The Three Curcuminoids== --&gt;<br /> <br /> <br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> <br /> <br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ===Flash Chromatography Data=== <br /> <br /> We used a 15.5mL C18 Column to run 1mL of our Curcumin extract. This column uses acetyl nitrile and water to run the liquid phase through the column. We ran a gradient for 25 min and got three distinct peaks which we then proceeded to run an HPLC method on.<br /> <br /> ===Sourcing turmeric===<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ===Overview of Proposed Methodology===<br /> ''Procedure is modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> <br /> *Extract curcuminoids with MeOH under soxhlet extraction.<br /> *Partition MeOH supernatant with ethyl acetate (AcOEt) and water for purification purposes.<br /> *Fractionate AcOEt extract by silica gel column chromatography to yield AcOEt curcumin eluents. <br /> *Fractionate AcOEt eluents by silica gel TLC using chloroform: acetone = 8:1 as the solvent system. <br /> *Purify each eluent by removing organic solvent (?) and analyzing under HPLC.<br /> <br /> ===Extracting Curcuminoids under Soxhlet Distillation===<br /> *Weigh out 80.00g of turmeric powder with an analytical balance. <br /> *Measure out 300.00mL MeOH using a burette. <br /> *Place above compound and solvent in stillpot of soxhlet apparatus. <br /> *Carefully turn on heat source and water source. <br /> *Run extraction for 6-7 hours.<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> <br /> ===3. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===4. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===5. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===6. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===7. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===8. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Curcumin_Research&diff=8856 2018-05-10T23:14:55Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Curcumin Research Initiative== --&gt;<br /> &lt;!-- ==The Three Curcuminoids== --&gt;<br /> &lt;!-- ==Flash Chromatography Data== --&gt;<br /> &lt;!-- ==NMR Data== --&gt;<br /> &lt;!-- ==Analyses and Discussion== --&gt;<br /> &lt;!-- ==Looking Ahead== --&gt;<br /> &lt;!-- --&gt;<br /> &lt;!-- --&gt;<br /> <br /> You have reached the page dedicated to the research of curcumin, a secondary plant metabolite and biophenol of interest to the Sturgeon Research Project. This page was created by [[Stehanie_Saey|Stephanie Saey]] and Nadia Ayala. Nadia was a 2017 Biochemistry graduate and Stephanie was a 2018 Biochemistry/Biopsychology graduate.<br /> <br /> ==Curcumin Research Initiative==<br /> <br /> ===Preparation of Curcuminoid Standards from Turmeric Plant===<br /> Curcumin is a secondary plant metabolite of the turmeric herb ''Curcuma longa''. The term &quot;curcumin&quot; has been used to refer to the bioactive molecule, but in reality curcumin has three derivatives (I/II/III) of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Together, the aforementioned compounds are known as curcuminoids. A review of literature suggests that curcuminoids have chemotherapeutic, antioxidant, and anti-inflammatory activity, among other uses to be discovered. However, difficulty arises when seeking to study these curcuminoids individually. Curcumin I is commercially available (10 mg for $150), while demethoxycurcumin (II) and bisdemethoxycurcumin (III) are not commercially available. Our current research project aims to isolate (methanol in soxhlet), purify (automated Flash/HPLC), and characterize (NMR) the curcuminoids for further investigation.<br /> <br /> ===The Three Curcuminoids===<br /> Curcumin I (Main Curcuminoid, in picture below), demethoxycurcumin (curcumin II), and bisdemethoxycurcumin (curcumin III) are the three major curcuminoids present in turmeric and of interest to this study. Their respective chemical structures are depicted below:<br /> <br /> [[Media:Curcuminoid Analogs with Potent Activity.pdf|Curcuminoid Analogs with Potent Activity (Article)]]<br /> <br /> [[File:Curcumin.jpg|400px|thumb|none|Curcumin I, II, and III]]<br /> <br /> ===Flash Chromatography Data=== <br /> <br /> We used a 15.5mL C18 Column to run 1mL of our Curcumin extract. This column uses acetyl nitrile and water to run the liquid phase through the column. We ran a gradient for 25 min and got three distinct peaks which we then proceeded to run an HPLC method on.<br /> <br /> ===Sourcing turmeric===<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> ===Overview of Proposed Methodology===<br /> ''Procedure is modeled after the following study:'' Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. Chem Pharm Bull (Tokyo) 1993;41:1640-3. <br /> <br /> *Extract curcuminoids with MeOH under soxhlet extraction.<br /> *Partition MeOH supernatant with ethyl acetate (AcOEt) and water for purification purposes.<br /> *Fractionate AcOEt extract by silica gel column chromatography to yield AcOEt curcumin eluents. <br /> *Fractionate AcOEt eluents by silica gel TLC using chloroform: acetone = 8:1 as the solvent system. <br /> *Purify each eluent by removing organic solvent (?) and analyzing under HPLC.<br /> <br /> ===Extracting Curcuminoids under Soxhlet Distillation===<br /> *Weigh out 80.00g of turmeric powder with an analytical balance. <br /> *Measure out 300.00mL MeOH using a burette. <br /> *Place above compound and solvent in stillpot of soxhlet apparatus. <br /> *Carefully turn on heat source and water source. <br /> *Run extraction for 6-7 hours.<br /> <br /> ==Written Report==<br /> <br /> ===1. Descriptive information===<br /> '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''<br /> <br /> Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon<br /> <br /> Special thanks to Michael Prinsell and Broddie Davis <br /> <br /> Research work documented in lab notebooks. <br /> <br /> ===2. Abstract ===<br /> Turmeric, Curcuma longa, is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties. The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives of different molecular structures: curcumin (I), demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography. However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids (I/II/III) in amounts large enough for further investigation on its radicalisation properties.<br /> ===3. Introduction===<br /> <br /> Turmeric, ''Curcuma longa,'' is a traditional Indian spice with potential chemotherapeutic, pharmacological, anti-inflammatory, and antioxidative properties (Goel, 2008). The active component in turmeric, known as curcumin, is what allows the plant to house its well-documented health benefits. Curcumin has three derivatives (I/II/III) of different molecular structures: curcumin I, demethoxycurcumin (II), and bisdemethoxycurcumin (III). Taken together, these structures are referred to as curcuminoids. Due to the safety and availability of turmeric, many studies have reported techniques for isolating and purifying the curcuminoids through methods such as extraction coupled with column chromatography (Jayaprakasha). However, to date, no such methods have been used to prepare large amounts of each curcuminoid individually. Curcumin I is only available in small amounts, while II and III remain unavailable commercially. The current research project aims to successfully isolate (methanol in soxhlet), purify (flash chromatography/HPLC), and characterize (NMR) the curcuminoids in amounts large enough for further investigation.<br /> <br /> ===4. Background from earlier reports===<br /> Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric. <br /> <br /> ===5. Experimental===<br /> :'''Sourcing turmeric:'''<br /> In order to proceed with the proposed research project, we needed to purchase turmeric from a reliable, science-conscious company. We chose to order 1 lb of organic and 1 lb of non-organic turmeric from Starwest Botanicals [http://www.starwest-botanicals.com/ here].<br /> <br /> :'''Methanol in Soxhlet Extraction:'''<br /> ::1. Soxhlet apparatus was set up for extraction, as pictured in Image A. <br /> ::2. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.<br /> ::3. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added. <br /> ::4. Heat source and water source were turned on. <br /> ::5. Extraction was ran for 6 hours. <br /> <br /> [[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]<br /> <br /> :'''Removal of MeOH and Impurities:'''<br /> ::1. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.<br /> ::2. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution. <br /> ::3. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute. <br /> ::4. The stopcock was removed, allowing the mixture to separate into two separate phases, one containing EtOAc and curcuminoids, and the other containing MeOH, water, and impurities. Image B displays this separated mixture. <br /> ::5. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container. <br /> ::6.The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime. <br /> ::7. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.<br /> ::8. The stopcock was removed and the mixture was allowed to separate. <br /> ::9. The top layer was added to the labeled glassware and the bottom layer was discarded. <br /> [[File:separatoryfunnel.jpg|200px|thumb|left|Image B: Separation of EtOAc and curcuminoids (top layer) from MeOH and impurities (bottom layer)]]<br /> <br /> :'''Thin Layer Chromatography'''<br /> ::A 5x10cm RediSep silica TLC plate was used to run TLC on the EtOAc/curcuminoid extract. First, 45 mL of the extract was diluted in 25 mL of ethyl acetate. A thin pencil line was drawn horizontally approximately 1.5 cm from the bottom of the plate. Two dots of the diluted extract were placed 4 cm apart on this line in order to conduct two trials on the same plate. An 8:1 chloroform:acetone solvent mixture was used. The fractions were allowed to follow the solvent up the plate and the results were imaged as shown in Figure A. Final Rf values were recorded. <br /> <br /> :'''Flash Column Chromatography:'''<br /> ::A 15.5mL(13g) gold C18 reverse phase Column was used to run 1mL of the EtOAc/curcuminoid extract. This column uses acetyl nitrile and acidic(TFA 0.1%) water to run the liquid phase through the column. <br /> :: A gradient was ran for 25 min that yielded three distinct peaks. The eluents for each peak were collected and stored in separate viles. <br /> <br /> :'''High Pressure Liquid Chromatography (HPLC):'''<br /> ::Each 1mL of each Flash eluent was added to a separate HPLC vile and labeled to be compared to the original Curcumin solution extracted through Soxhlet extraction. <br /> [[File:Curcumin HPLC vial image.jpg|400px|thumb|left|Image c: HPLC vials]]<br /> <br /> :'''Rotovaporization''':<br /> :: Each fraction of curcumin had its solvent extracted via rotovaportization. The hot bathwater was held at 32 degrees Celsius for about an hour and a half or until completely dry. Spin speed was between 9 and 10, and adjustments in the angle of inundation of the round bottom flask containing the curcumin fractions was adjusted as the solvent was vaporized.<br /> <br /> :'''Nuclear Magnetic Resonance (NMR) Testing:'''<br /> ::About 1mg of each curcumin was added to separate NMR tubes. 1mL of duderated DMSO was added to resuspend the curcumin samples to be used in the 400Hz NMR at Knox College. <br /> <br /> ===6. Results===<br /> ::'''TLC Results'''<br /> :TLC results from two trials of the diluted EtOAc/curcuminoid mixture (prepared from MeOH in soxhlet extraction) are shown in Figure A. A RediSep silica TLC plate was used with an 8:1 chloroform:acetone solvent front. Three distinct lines are visible for each trial, consistent with the three different curcuminoid structures. Rf values are as follows (same for each trial): .25, .375, .5.<br /> [[File:Example.jpg|200px|thumb|none|Figure A: TLC Results]]<br /> <br /> ::'''Flash Chromatography Results'''<br /> :With the use of a C18 reverse phase column we successfully separated Curcumin I, II, and III from the original solution. The base line resolution of the curcumin fractions wasadjusted by increasing the run time from the original C18 procedure from 15 min to 25 min. <br /> [[Media:Curcumin Flash graph.pdf|Curcumin Flash Chromatography Image]]<br /> <br /> ::'''HPLC Results'''<br /> :Having separated the three curcumin through flash chromatography, purity of the sample was tested using flash chromatography using the original solution as a basis of comparison.<br /> [[File:T2 Curcumin HPLC chromatogram.jpg|500px|thumb|none|HPLC chromatogram]]<br /> <br /> ::'''NMR Results'''<br /> :The concentration of our curcumins where not significant enough to have transmitted a high signal in the NMR for what we precive to be curcumin II and Curcumin III. However, the NMR results for Curcumin II are seen below.<br /> [[File:Curcumin 1 NMR.jpg|500px|thumb|none|NMR Curcumin I Trial 1]]<br /> <br /> ===7. Discussion===<br /> The flash chromatography and HPLC data have been consistent with the literature regarding the existence of curcumin's three different molecular structures. However, characterization of curcumins through NMR have led to further questions on the structures present within each curcumin sample. Further interpretation and testing are been necessary to have certainty in the purity of our samples as well as specific characterization of each fraction.<br /> <br /> ===8. Conclusions===<br /> No final conclusions have been reached at this point, as we are still waiting to collect NMR results. The results of the NMR should tell us how successful our method was in isolating and purifying curcuminoids I,II, and III.<br /> <br /> ===9. Future Directions===<br /> We plan to continue this research project into the second semester of the 2016-17 school year (and beyond). First on our agenda will be to collect NMR data on the three separate curcuminoids that were rotovapped from the separate flash eluents. NMR data will be collected both at Monmouth College and Knox College. Results will indicate the success of our methods, and lead us to either additional method development or mass quantification using the latest method.<br /> <br /> ===10. Literature references===<br /> <br /> Goel A., Kunnumakkara A.B., Aggarwal B.B. (2008). Curcumin as ‘curecumin’: from kitchen to clinic. ''Biochem Pharmacology.'' pp. 787–809, doi:10.1016/j.bcp.2007.08.016 <br /> <br /> Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., &amp; Patil, B. S. (2013). Rapid separation and quantitation of curcuminoids combining pseudo two dimensional liquid flash chromatography and NMR spectroscopy. ''Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences'', pp. 937, doi:10.1016/j.jchromb.2013.08.011<br /> <br /> Kiuchi, F, Goyto, Y, Sugimoto, N, Akao, N, Kondo, K, Tsuda, Y. Nematocidal activity of turmeric: synergistic action of curcuminoids. (1993). ''Chem Pharm Bull (Tokyo)'' pp. 1640-3.<br /> <br /> ===10. Signature===<br /> Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8855 2018-05-10T23:07:18Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by [[Stephanie_Saey|Stephanie Saey]]<br /> Please visit the following link for an updated report: [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8854 2018-05-10T23:07:07Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by [[Stephanie_Saey|Stephanie Saey]]<br /> Please visit the following link for an updated report:[[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8853 2018-05-10T23:06:35Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by [[Stephanie_Saey|Stephanie Saey]]<br /> [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8852 2018-05-10T23:05:57Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala; continued into Summer 2017 by Stephanie Saey<br /> [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Stephanie_Saey&diff=8851 2018-05-10T23:05:07Z

<p>Ssaey: </p> <hr /> <div>&lt;!-- ==Personal Information== --&gt;<br /> &lt;!-- ==Undergraduate Research Activities== --&gt;<br /> &lt;!-- ==Presentations== --&gt;<br /> &lt;!-- ==Publications== --&gt;<br /> &lt;!-- ==Activities== --&gt;<br /> &lt;!-- ==Career Goals== --&gt;<br /> &lt;!-- --&gt;<br /> &lt;!-- --&gt;<br /> You have reached the personal page of Stephanie Saey. <br /> <br /> ==Personal Information==<br /> Graduated (Spring 2018) Biochemistry and Biopsychology Double Major <br /> <br /> Hometown: Galesburg, Illinois<br /> <br /> ==Undergraduate Research Activities==<br /> <br /> Summer 2015: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Kristin Larson <br /> *Reviewed literature surrounding the psychological phenomenon of the &quot;third place&quot;<br /> *Developed a research proposal to study the relationship between geographical setting of the &quot;third place&quot; coffee shop and individual place attachment<br /> *Wrote HSRB and developed survey<br /> *Collected data in Chicago, Illinois, and Galesburg, Illinois<br /> *Analyzed data using Statistical Software Minitab<br /> <br /> Summer 2016: Freeman Grant Recipient, Travelled to Singapore for research with Dr. Kristin Larson <br /> *Collaborated with Dr. Kristin Larson and two undergraduate colleagues to develop a research proposal to study in Singapore <br /> *Submitted the research proposal to the ASIANetwork Student-Faculty Fellows Program <br /> *Received the ASIANetwork grant, funded by the Freeman Foundation, to study for three weeks in Singapore <br /> *Wrote HSRB and developed survey to study the relationship between &quot;third places&quot; and perceived stress levels <br /> *Currently (Fall 2016 semester) in the process of analyzing data and writing a post-report <br /> <br /> Summer 2016: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Bradley Sturgeon and Dr. Laura Moore <br /> *Served as a student mentor for incoming freshmen participating in the three-week program<br /> *Led discussions and small experiments on the chemistry of basic baking (i.e., gluten formation and yeast activity)<br /> *Performed an extraction of gluten proteins from various types of flour dough and analyzed the extractions through SDS-PAGE <br /> *Mentored the students in developing a presentation for the SOfIA colloquium <br /> *Helped supervise the making of final posters for the project <br /> <br /> Fall 2016-Spring 2017: Chem 430 with B. Sturgeon, worked as a mentee of Nadia Ayala<br /> Spring 2017: Continued research with the secondary plant metabolite, curcumin:<br /> [[Curcumin Research|Curcumin Research]]<br /> Fall 2017-Spring 2018: Worked to oxidize lignin monomers (mentee: Zelinda Taylor) <br /> ==Presentations==<br /> <br /> August 2015: '''SOfIA Presentation'''<br /> <br /> Poster Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> Spring 2016: '''ILLOWA Undergraduate Research Conference''' Macomb, IL<br /> <br /> Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> August 2016: '''SOfIA Presentation'''<br /> <br /> Poster Titles: ''The Chemistry of Baking: The Basics, The Chemistry of Baking: Experiments''<br /> <br /> <br /> ==Publications==<br /> Spring 2016: '''The 2016-17 Midwest Journal of Undergraduate Research'''<br /> <br /> Publication Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> ==Activities==<br /> <br /> <br /> ==Career Goals==<br /> <br /> Psychiatry</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Stephanie_Saey_Chem430_F16&diff=8850 2018-05-10T23:02:16Z

<p>Ssaey: Replaced content with &quot;Please see Curcumin Research for a detailed report of this student's work during the fall of 2016.&quot;</p> <hr /> <div>Please see [[Curcumin_Research|Curcumin Research]] for a detailed report of this student's work during the fall of 2016.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Biophenols_as_Secondary_Plant_Metabolites&diff=8849 2018-05-10T22:56:51Z

<p>Ssaey: </p> <hr /> <div>==[https://en.wikipedia.org/wiki/Lignin Lignin] monomers==<br /> [[File:Ligin Monomer radicals.png|200px|thumb|center|ESR data from HRP oxidation of lignin monomers]]<br /> :[https://en.wikipedia.org/wiki/Paracoumaryl_alcohol p-coumaryl alcohol]<br /> :[https://en.wikipedia.org/wiki/Coniferyl_alcohol coniferyl alcohol]<br /> :[https://en.wikipedia.org/wiki/Sinapyl_alcohol sinapyl alcohol]<br /> <br /> ==Lignans (General)==<br /> <br /> ==Eugenol/Methyl Eugenol/Methyl Chavicol (from Clove or Basil)==<br /> :Previous work by [http://esr.monmsci.net/wiki/index.php/Pauzi_Research_Spring_2016 Matt (Amad Pauzi)]<br /> :Current (Fall 2016) work by [http://esr.monmsci.net/wiki/index.php/Lang_Research_2016 Sarah Lang].<br /> <br /> [[Enzymatic_oxidation_of_Methyl_Eugenol_and_Methyl Chavicol|Enzymatic oxidation of Methyl Eugenol and Methyl Chavicol]]<br /> <br /> ==Curcumin (from Turmeric)==<br /> :Project started (Fall 2016) by Stephanie Saey and Nadia Ayala.<br /> [[Curcumin_Research|Curcumin Research]]<br /> :[[Media:Turmeric ref 01.pdf|Reference]]; purification from plant material.<br /> <br /> ==Chlorogenic Acid (from green coffee beans)==<br /> :Project started by [[Chris_Knutson]].<br /> :Current project on isolation from green coffee beans by Alan Kuhlemier.<br /> <br /> References<br /> :[[Media:Jf9605254.pdf|Five ways to Isolate Chlorogenic acid.]]<br /> :[[Media:Isolation microwave.pdf|Microwave assisted extraction with other references to standard solvent extractions.]]<br /> <br /> ==Working with Plant Material to Recover Biophenols==<br /> ===Essential Oils From Basil===<br /> Project starting the summer 2017, but additional work on &quot;steam distillation&quot; was started in Fall 2016 by Ben Stillwell and Selene H.<br /> <br /> [http://esr.monmsci.net/wiki/index.php/Extraction_and_Characterization_of_Essential_Oils_from_Basil_Varieties Extraction and Characterization of Essential Oils from Basil Varieties]<br /> <br /> References:<br /> :[[Media:17688.pdf|Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia]]<br /> :[[Media:Grayer 1996.pdf|Infraspecific Taxonomy and Essential Oil Chemotypes in Sweet Basil, ''Ocimum basilicum''.]]<br /> <br /> ===Extraction of Flavonoids from Basil===<br /> [[Media:Grayer 1996b.pdf|EXTERNAL FLAVONES IN SWEET BASIL, OCIMUM BASILICUM, AND RELATED TAXA]]<br /> <br /> ===Extractions of Witch Hazel===<br /> https://en.wikipedia.org/wiki/Witch_hazel_(astringent)</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Stephanie_Saey&diff=8848 2018-05-10T22:51:30Z

<p>Ssaey: /* Personal Information */</p> <hr /> <div>&lt;!-- ==Personal Information== --&gt;<br /> &lt;!-- ==Undergraduate Research Activities== --&gt;<br /> &lt;!-- ==Presentations== --&gt;<br /> &lt;!-- ==Publications== --&gt;<br /> &lt;!-- ==Activities== --&gt;<br /> &lt;!-- ==Career Goals== --&gt;<br /> &lt;!-- --&gt;<br /> &lt;!-- --&gt;<br /> You have reached the personal page of Stephanie Saey. <br /> <br /> ==Personal Information==<br /> Graduated (Spring 2018) Biochemistry and Biopsychology Double Major <br /> <br /> Hometown: Galesburg, Illinois<br /> <br /> ==Undergraduate Research Activities==<br /> <br /> Summer 2015: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Kristin Larson <br /> *Reviewed literature surrounding the psychological phenomenon of the &quot;third place&quot;<br /> *Developed a research proposal to study the relationship between geographical setting of the &quot;third place&quot; coffee shop and individual place attachment<br /> *Wrote HSRB and developed survey<br /> *Collected data in Chicago, Illinois, and Galesburg, Illinois<br /> *Analyzed data using Statistical Software Minitab<br /> <br /> Summer 2016: Freeman Grant Recipient, Travelled to Singapore for research with Dr. Kristin Larson <br /> *Collaborated with Dr. Kristin Larson and two undergraduate colleagues to develop a research proposal to study in Singapore <br /> *Submitted the research proposal to the ASIANetwork Student-Faculty Fellows Program <br /> *Received the ASIANetwork grant, funded by the Freeman Foundation, to study for three weeks in Singapore <br /> *Wrote HSRB and developed survey to study the relationship between &quot;third places&quot; and perceived stress levels <br /> *Currently (Fall 2016 semester) in the process of analyzing data and writing a post-report <br /> <br /> Summer 2016: Summer Opportunity for Intellectual Activity (SOfIA) with Dr. Bradley Sturgeon and Dr. Laura Moore <br /> *Served as a student mentor for incoming freshmen participating in the three-week program<br /> *Led discussions and small experiments on the chemistry of basic baking (i.e., gluten formation and yeast activity)<br /> *Performed an extraction of gluten proteins from various types of flour dough and analyzed the extractions through SDS-PAGE <br /> *Mentored the students in developing a presentation for the SOfIA colloquium <br /> *Helped supervise the making of final posters for the project <br /> <br /> Fall 2016: Chem 430 with B. Sturgeon<br /> *Currently in process<br /> [[Curcumin Research|Curcumin Research]]<br /> [[Stephanie Saey Chem430 F16| Updated Curcumin Research]]<br /> <br /> ==Presentations==<br /> <br /> August 2015: '''SOfIA Presentation'''<br /> <br /> Poster Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> Spring 2016: '''ILLOWA Undergraduate Research Conference''' Macomb, IL<br /> <br /> Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> August 2016: '''SOfIA Presentation'''<br /> <br /> Poster Titles: ''The Chemistry of Baking: The Basics, The Chemistry of Baking: Experiments''<br /> <br /> <br /> ==Publications==<br /> Spring 2016: '''The 2016-17 Midwest Journal of Undergraduate Research'''<br /> <br /> Publication Title: ''The Third Place Experience in Urban and Rural Coffee Shops''<br /> <br /> <br /> ==Activities==<br /> <br /> <br /> ==Career Goals==<br /> <br /> Psychiatry</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7788 2017-09-07T18:08:40Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.jpg|200px|thumb|left|Image A: Insoles before Run]]<br /> <br /> [[File:AfterRun.jpg|200px|thumb|left|Image B: Insoles after Run]]<br /> <br /> <br /> I chose to take pictures of the insoles of my running shoes both before (Image A) and after running (Image B). Running is something I do everyday; I thought it would be interesting to see how much heat transfer occurs from my body to my shoes through the running process, as well as which areas of my foot are utilized the most. As is noticeable in Image A, the insoles of my shoes before running were around 68-70 degrees Fahrenheit (corresponding to the large yellow/green areas). Surprisingly, the heels of my insoles were warmer, producing a red image corresponding to approximately 71 degrees Fahrenheit. I'm not sure why this was the case for the insoles &quot;at rest;&quot; because I hadn't used my shoes in over 12 hours, I expected the insoles to be the same temperature throughout in the &quot;Before&quot; picture. <br /> Image B shows my insoles after running, during which the overall temperature of the insoles increased to be around 75 degrees. However, the middle part of my feet around the area of my arches was the hotter than the heel area (~80 degrees F versus 75 degrees F). I attribute this to mid-foot striking. When I run, I strike the ground mid-foot, allowing the area to be more weight-bearing; thus, my feet are in greater contact with the insoles around my mid-foot.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7787 2017-09-07T18:08:16Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.jpg|200px|thumb|left|Image A: Insoles before Run]]<br /> <br /> [[File:AfterRun.jpg|200px|thumb|left|Image B: Insoles after Run]]<br /> <br /> <br /> I chose to take pictures of the insoles of my running shoes both before (Image A) and after (Image B) running. Running is something I do everyday; I thought it would be interesting to see how much heat transfer occurs from my body to my shoes through the running process, as well as which areas of my foot are utilized the most. As is noticeable in Image A, the insoles of my shoes before running were around 68-70 degrees Fahrenheit (corresponding to the large yellow/green areas). Surprisingly, the heels of my insoles were warmer, producing a red image corresponding to approximately 71 degrees Fahrenheit. I'm not sure why this was the case for the insoles &quot;at rest;&quot; because I hadn't used my shoes in over 12 hours, I expected the insoles to be the same temperature throughout in the &quot;Before&quot; picture. <br /> Image B shows my insoles after running, during which the overall temperature of the insoles increased to be around 75 degrees. However, the middle part of my feet around the area of my arches was the hotter than the heel area (~80 degrees F versus 75 degrees F). I attribute this to mid-foot striking. When I run, I strike the ground mid-foot, allowing the area to be more weight-bearing; thus, my feet are in greater contact with the insoles around my mid-foot.</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7780 2017-09-07T17:09:55Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.jpg|200px|thumb|left|Image A: Insoles before Run]]<br /> <br /> [[File:AfterRun.jpg|200px|thumb|left|Image A: Insoles after Run]]<br /> <br /> <br /> I chose to</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:AfterRun.jpg&diff=7779 2017-09-07T17:06:12Z

<p>Ssaey: Ssaey uploaded a new version of File:AfterRun.jpg</p> <hr /> <div></div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:AfterRun.jpg&diff=7778 2017-09-07T17:05:46Z

<p>Ssaey: </p> <hr /> <div></div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7777 2017-09-07T17:04:59Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.jpg|200px|thumb|left|Image A: Insoles before Run]]<br /> <br /> [[File:AfterRun.jpg|200px|thumb|left|Image A: Insoles after Run]]</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:BeforeRun.jpg&diff=7773 2017-09-07T15:08:07Z

<p>Ssaey: </p> <hr /> <div></div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7772 2017-09-07T15:06:48Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.jpg|200px|thumb|left|Image A: Insoles before Run]]</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7771 2017-09-07T15:04:57Z

<p>Ssaey: </p> <hr /> <div>[[File:File:BeforeRun.png|200px|thumb|left|Image A: Insoles before Run]]</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7770 2017-09-07T15:04:35Z

<p>Ssaey: </p> <hr /> <div>[[File:BeforeRun.png|200px|thumb|left|Image A: Insoles before Run]]</div>

Ssaey http://205.166.159.208/wiki/index.php?title=Thermal_Running&diff=7769 2017-09-07T15:03:35Z

<p>Ssaey: Created page with &quot;Image A: soxhlet extraction&quot;</p> <hr /> <div>[[File:Soxhlet.png|200px|thumb|left|Image A: soxhlet extraction]]</div>

Ssaey http://205.166.159.208/wiki/index.php?title=File:AfterRun.JPG&diff=7768 2017-09-07T15:02:05Z

<p>Ssaey: </p> <hr /> <div></div>

Ssaey