Source: UNIV OF MISSISSIPPI submitted to NRP
HIGH PERFORMANCE LIQUID CHROMATOGRAPH FOR RESEARCH ON UTILIZATION OF NATURAL PRODUCTS IN AGRICULTURE AND FORESTRY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
NRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
0194441
Grant No.
2003-35504-12877
Cumulative Award Amt.
(N/A)
Proposal No.
2002-01735
Multistate No.
(N/A)
Project Start Date
Nov 15, 2002
Project End Date
Nov 14, 2003
Grant Year
2003
Program Code
[71.2]- (N/A)
Recipient Organization
UNIV OF MISSISSIPPI
(N/A)
UNIVERSITY,MS 38677
Performing Department
PHARMACOGNOSY
Non Technical Summary
Natural products have a great potential to be valuable materials for future development into biologically active agents useful in agriculture and forestry. Many of them already show promise as environmentally friendly insecticides, herbicides and fungicides. Plants and marine invertebrates have also been shown to be a very rich source of compounds possessing antioxidant activities. Fungicidal, insecticidal, and antioxidant activities of some natural products make them also good candidates for potential use as wood protective agents against fungi termites. The crude extracts of plants, marine organisms, and fungi comprise a complex mixture of organic compounds from which the individual biologically active components are isolated by means of chromatography. High Performance Liquid Chromatography (HPLC) is one of the most efficient methods of separating complex mixtures of structurally related compounds, which are often encountered in natural product chemistry. HPLC systems are fundamental components of any natural products isolation program and therefore absolutely essential for the successful development of new agents for agriculture and forestry. The use of HPLC instrument will not only significantly facilitate the research in this field but also will enhance the likelihood of funding new projects. In addition the HPLC instrument will play an important role in the teaching process of undergraduate and graduate students.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
5112499200034%
5113899200033%
5114020200033%
Knowledge Area
511 - New and Improved Non-Food Products and Processes;

Subject Of Investigation
3899 - Other animals, general; 4020 - Fungi; 2499 - Plant research, general;

Field Of Science
2000 - Chemistry;
Goals / Objectives
The primary goal of this project is to purchase and utilize high performance liquid chromatograph (HPLC) in the research on discovery and development of natural products that can be beneficial for agriculture and forestry. HPLC instrument will be utilized to separate mixtures of secondary metabolites present in the crude extracts of selected plants, marine organisms and fungi from our own collection in order to find the most active compounds useful in the existing projects. This includes the identification of new hervicides, fungicides, and novel, prototype wood-preservatives to overcome the serious short-comings of existing products. HPLC instrument will also be used to analyze the purity of identified prototypes prior to their biological evaluation. In long terms HPLC instrument will be utilized in the process of discovery and development of novel natural products and their semi-synthetic analogs as useful materials for agriculture and forestry. This will involve the multi-personnel use of the instrument in the department. The instrument will also be used in the teaching process of undergraduate and graduate students. The separation of secondary metabolite mixtures is the major part of the departmental curriculum and hands-on course will improve students knowledge of high-performance liquid chromatography. Finally, HPLC instrument will allow for rapid collection of preliminary data for submission of competitive standard research grants.
Project Methods
The crude extracts of plants, marine organisms, and fungi comprise a complex mixture of organic compounds from which the individual biologically active components are isolated by means of chromatography. High Performance Liquid Chromatography (HPLC) is one of the most efficient methods of separating complex mixtures of structurally similar or related compounds, which are very often encountered in natural product chemistry. Natural products are frequently isolated following the evaluation of a relatively crude extract in a biological assay in order to fully characterize the active entity. In this setting, the requirement for speed will dictate that rapid, sscaleable, high-resolution techniques such as preparative HPLC are the most usefully applied very early in an extraction process so that the first milligrams of an active compound are provided in a timely manner. The biologically active entity is often present only as a minor component in the extract, and the resolving power of HPLC is ideally suited to the rapid processing of such multicomponent samples on both an analytical and preparative scale. Larger scale isolations are often required in order to provide more of a metabolite of interest for further biological or chemical studies. When operating at scales greater than the origianl isolation, many additional metabolites related to the principal component of interest (minor metabolites) may be revealed, and preparative HPLC is often an ideal technique to apply to their isolation. HPLC has been applied to the isolation of virtually all classes of natural products of plant, marine, and microbial origin; separations that were considered extremely challenging a few decades ago are now often relatively routine, and in that time, HPLC has become an indispensable tool for most products chemistry laboratories.

Progress 11/15/02 to 11/14/03

Outputs
The major goal of this project was to purchase and utilize high performance liquid chromatograph (HPLC) in the research on discovery and development of natural products that can be beneficial for agriculture and forestry. On November 15, 2002 I was awarded $25,000 from United States Department of Agriculture for purchasing such instrument. This grant was awarded from NRICGP-AREA-Equipment Grant (80.2) program for the non-food characterization product research (71.2). Together with matching money ($17,000) granted to me by the Associates Fund of University of Mississippi I was able to purchase a modern High Performance Liquid Chromatograph (HPLC) from the world-best manufacturer of such instrumentation, Waters Corporation in Milford, MA. On February 2003 semi-preparative Waters Delta Prep 4000 HPLC system was installed in my laboratory at the Department of Pharmacognosy of the University of Mississippi. This system includes PrepLC Controller, PrepLC 4000 Series Solvent Delivery Unit, 7725i Injector, Waters 2487 Dual Wavelength Absorbance Detector, 2487 Semi-Prep Taperslit Cell, and Empower Personal Preconfigured Single System Workstation for Windows XP. After a training session from Waters professional we started to use our HPLC in everyday laboratory practice. This instrument proved to be very useful in all our research projects especially in those related to utilization of natural products in agriculture and forestry. It significantly helped to identify new compounds with antifungal activities and also to study secondary metabolites from various wood-decaying fungi. The purchased Waters Delta Prep 4000 HPLC system is a versatile instrument allowing for analytical as well preparative separation of complex mixtures and proved to especially useful in the natural product research by utilizing a normal or reverse phase chromatographic columns. In the period of the duration of this grant (i.e. from the purchase of HPLC in February, 2003 to November 15, 2003) this instrument was extensively used in my research group by staff researchers and students in several projects resulting in four publications. All short-term goals formulated in this project were achieved, including its use for separation of the individual biologically active components from the crude extracts of plants, marine organisms, and fungi, and analyzing the purity of natural products and synthetic compounds prior to their biological evaluations for activities useful in agriculture and forestry. Several natural products and synthetic compounds with herbicidal and antifungal activity were found thanks to the use of this instrument. The work to achieve long-term goals to utilize HPLC instrument in the discovery and development of new products for agriculture and forestry is currently in progress. It involves the collection of preliminary data for submission of competitive standard research grants, multi-personnel use of the instrument and teaching of undergraduate and graduate students in proper use of this instrument.

Impacts
The purchase of HPLC instrument had a great impact on efficiency and effectiveness of our research in natural products chemistry. It has an economic impact on our research, saving time and money, allowing for faster separation of the complex mixtures of secondary metabolites without necessity of use of large quantities of traditional chromatographic materials. It has also a positive environmental impact by eliminating the need of disposal of large quantities of used silica gel otherwise used in absorption column chromatography, and by reducing the amount of organic solvents used in the process.

Publications

  • Fahmy, H., Zjawiony, J.K., Khalifa, S., and Fronczek, F. 2003. A Semisynthetic Analog of the Cembranoid Sarcophine. Acta Cryst. C59, o85-o87.
  • Gochfeld, D.J., El Sayed, K.A., Yousaf, M., Hu, J.F., Bartyzel, P., Dunbar, D.C., Wilkins, S.P., Zjawiony, J.K., Schinazi, R.F., Schlueter-Wirtz, S.P., Tharnish, M., and Hamann, M.T. 2003. Marine Natural Products as Lead Anti-HIV Agents. Mini Rev. Med. Chem. 3, 401-425.
  • Fahmy, H., Rostom, S., Saudi, M.N., Zjawiony, J.K., and Robins, D.J. 2003. Synthesis and in-vitro evaluation of the anticancer activity of novel fluorinated thiazolo[4,5-d]pyrimidines. Arch. Pharm. Pharm. Med. Chem. 336, 216-225.
  • Katsuyama, I., Khalil, A.A., Dunbar, C., and Zjawiony, J.K. 2003. Concentration-Dependent Variation in 1H-NMR Chemical Shifts of Aromatic Protons in Sampangine Derivatives. Spectroscopy Letters. 36(5 & 6), 473-481.