Difference between revisions of "Stephanie Saey Chem430 F16"

Latest revision as of 23:02, 10 May 2018

Please see Curcumin Research for a detailed report of this student's work during the fall of 2016.

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-Chemistry/Biochemistry Research 430
+Please see [[Curcumin_Research|Curcumin Research]] for a detailed report of this student's work during the fall of 2016.
-:Fall 2016
-:Stephanie Saey
-:Junior Biochemistry and Biopsychology Double Major
-==Research Times==
-Mon: 12-2pm
-Th: 11-1pm
-: section 01 = 0.25 credit = 4 hours per week.
-==Proposed Research Project==
-===General Information===
-:Advisor: Dr. Brad Sturgeon
-:Other research student collaborators: Nadia Ayala
-=== Proposal: Preparation of Curcuminoid Standards from Turmeric Plant===
-Curcumin is a secondary plant metabolite of the turmeric herb ''Curcuma longa.'' The term "curcumin" has been used to refer to the bioactive molecule, but in reality curcumin has three derivatives of different molecular structures: curcumin I, demethoxycurcumin (II), and bisdemethoxycurcumin (III). Together, these structures are known as curcuminoids. A review of curcumin studies suggest curcuminoids have chemotherapeutic, antioxidant, and anti-inflammatory activity, among other uses to be discovered.  However, difficulty arises when seeking to study the derivatives of curcumin individually.  Curcumin I is only available in small amounts, while demethoxycurcumin and bisdemethoxycurcumin remain unavailable commercially.  The current research project aims to successfully isolate, purify, and characterize the curcuminoids in amounts large enough for further investigation. Using MeOH and a soxhlet apparatus, the curcuminoids will be extracted from turmeric and subsequently subjected to Flash Chromatography for separation. Once the curcuminoids are separated, NMR data collection will take place to compare the chemical properties of each derivative.  The process will be repeated multiple times to prepare substantial standards of the curcuminoids.
-===Instruments to be used===
-Distillation Apparatus, Flash Chromotography System, NMR
-===References===
-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
-Jayaprakasha, G. K., Gowda, G. A. N., Marquez, S., & 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
-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.
-===Research pledge===
-I, Stephanie Saey, have read the Chem/Bioc 430 course syllabus and understand the general structure and expectations of the research program. The above material was prepared after consultation, and in conjunction with my research advisor, Dr. Brad Sturgeon.
-==Written Report==
-===1. Descriptive information===
-'''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''
-Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon#
-Research work documented in lab notebooks, Stephanie Saey pages 1-13 and Nadia Ayala pages [insert NA page#].
-===2. Introduction===
-Turmeric, ''Curcuma longa,'' is a traditional Indian spice with potential chemotherapeutic, pharmacological, and 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 its good 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.
-===3. Background from earlier reports===
-Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric.
-===4. Experimental===
-:''Sourcing turmeric:''
-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].
-:''Methanol in Soxhlet Extraction:''
-. Soxhlet apparatus was set up for extraction, as pictured below.
-. Approximately 76 grams of organic turmeric from Starwest Botanicals was added to the thimble.
-. Approximately 350 mL of MeOH was added to the stillpot and a few boiling stones were added.
-. Heat source and water source were turned on.
-. Extraction was ran for 6 hours.
-:''Removal of MeOH and Impurities:''
-. MeOH/curcuminoid mixture in the stillpot (from extraction) was transferred to an 1000mL separatory funnel.
-. 275 mL of ethyl acetate (EtOAc) and 150 mL of water were added to the funnel, along with 150 mL brine solution.
-. The stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.
-. 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.
-. The EtOAc/curcuminioid phase (top layer) was drained into a labeled glassware container.
-The bottom layer was readded to the separatory funnel along with 150 mL of EtAOc, 150 mL water, and 100 mL brime.
-. Again, the stopcock was added to the funnel and the mixture was inverted and vented multiple times for 1 minute.
-.
-::'''Reagents''': ''List reagents used by product number and the source''.
-::'''Enzyme Reactions''': All enzyme reactions were done using a totlal volume of 5.0 mL. Substrate concentration ranged between 1-12 mM. Reactions were initiated be the addition of enzyme.
-::'''HPLC''': HPLC data was collected using the Waters Breeze HPLC with a C18 column (insert specs here). Specific HPLC conditions are given in the figure captions.
-::'''UV-Vis''': UV-Vis data was collected using the HP 5832 UV-Vis spectrometer using a quartz cuvette. Sample concentrations were adjusted so as to not exceed 1.0 absorbance unit. Significant data was exported in the ".CSV" format and then worked up in Igor.
-===5. Results===
-This section documents the results of the experiments done during the period covered by the report.  Tabulation of data is encouraged; representative spectra presented.
-===6. Discussion===
-In this section, the student will discuss the results of their work in context of the literature and the results of earlier reports.  Questions that have been raised in earlier reports may be addressed here.
-===7. Conclusions===
-Restate the findings of this period of research. The statement “No conclusions have been reached,” is an acceptable statement.
-===8. Future Directions===
-In this section, the student will discuss possible experiments intended to address unanswered questions or technical problems on the project.
-===9. Literature references===
-Literature referenced in the report will be cited.  This will be copied from the earlier report, and the student is expected to contribute to the accumulation of relevant literature. Remember that all cited literature must be read.
-===10. Signature===
-Two copies of the report will be signed and dated and turned in to the Faculty Research Advisor and archived by the Research Coordinator.

Difference between revisions of "Stephanie Saey Chem430 F16"

Latest revision as of 23:02, 10 May 2018

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