Difference between revisions of "Stephanie Saey Chem430 F16"

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Chemistry/Biochemistry Research 430

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 here.

Example:

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.

@@ Line 17: / Line 17: @@
 :Other research student collaborators: Nadia Ayala
-=== Proposal: Preparation of Curcuminoid Standards from Tumeric Plant===
+=== 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, and bisdemethoxycurcumin. 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.
+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===
@@ Line 25: / Line 25: @@
 ===References===
-Goel A, Kunnumakkara AB, Aggarwal BB. Curcumin as ‘curecumin’: from kitchen to clinic.  Biochem Pharmacol. 2008;75:787–809.
+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
-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.
+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===
@@ Line 36: / Line 39: @@
 ===1. Descriptive information===
 '''Isolation and Purification of Curcuminoids from Tumeric Plant ''Curcuma Longa'''''
-A project title, the names of the researcher (past and current), faculty research advisor, and other relevant student researchers and/or collaborators, the name and page numbers of the lab notebook(s) where the research is described, the dates when the work was done, and the names of the document file and its immediate precursor.
-Example:
-'''Detection of Radical Intermediate generated from the MAO enzymatic system.'''
-Styles Bitchly*, Donald Hump, Penn Mickey, and Bradley E. Sturgeon#
+Stephanie Saey, Nadia Ayala, and Bradley E. Sturgeon#
-*lead author, #research advisor
+Research work documented in lab notebooks, Stephanie Saey pages 1-13 and Nadia Ayala pages [insert NA page#].
-Research work documented in lab notebooks, SB_01 pages 1-25.
 ===2. Introduction===
-State of the motivation for the project of interest in terms of current literature. When we make reference to current literature, this is where you provide specific reference(s) to this work. This section can be largely copied from an earlier report(s), if they exist, but it is expected that this section is edited to include newly found background information. Note: if the author of previous report(s) is included as a contributor to the project, you may cut and paste text; this is not plagiarism, it is collaboration.
+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===
-This section will summarize the work reported in earlier reports.  If significant results were presented in the most recent report, these results will be summarized and in most cases can be added to that reports background section. This section is included mainly to confirm the current students overall understanding of the project.
+Stephanie Saey and Nadia Ayala are the first Monmouth College Chemistry students to work on this specific research project involving turmeric.
 ===4. Experimental===
-This section describes experiments done during the period covered by the report.  Usually, this work will be similar or identical to that described in the prior report, if they exists.  Any new experiments will be written up and added to the experimental section. Any experiments preformed under identical conditions need only reference previous reports.
+===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].
 :''Example:''