Source: MINNESOTA STATE UNIVERSITY, MANKATO submitted to NRP
ACQUIRING A HIGH PERFORMANCE LIQUID CHROMATOGRAPHY SYSTEM WITH MASS SPECTRAL CAPABILITIES (LCMS)
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
0210878
Grant No.
2007-35200-18233
Cumulative Award Amt.
(N/A)
Proposal No.
2007-02291
Multistate No.
(N/A)
Project Start Date
Jul 15, 2007
Project End Date
Jul 14, 2008
Grant Year
2007
Program Code
[31.0]- (N/A)
Recipient Organization
MINNESOTA STATE UNIVERSITY, MANKATO
228 WIECKING CTR
MANKATO,MN 56001-6062
Performing Department
(N/A)
Non Technical Summary
1. Damage to certain essential chemicals in our bodies has been suggested to be related to age-related development of some cancers, development of cardiovascular disease, cataractogenesis, atherosclerosis, rheumatoid arthritis, diabetes, and neurodegenerative diseases including Alzheimer's disease. Compounds call antioxidants may help to reduce the damage. 2. Antibiotics that make their way into the soil can also appear in our food supply and/or contribute to an increase in antibiotic resistant organisms. 3. Light can make oxygen very reactive (photochemical processes). These photochemical processes strongly influence the active lifetime, bioavailability, degradation products, and ultimate fate of agrochemicals that are applied to soil surfaces. The purpose of this project is to purchase a liquid chromatography system with mass spectral capabilities.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70272991010100%
Goals / Objectives
The objective of this project is the purchase of a Shimadzu liquid chromatography system with mass spectral capabilities (LCMS)that will be housed in the Chemistry and Geology Department at Minnesota State University, Mankato.
Project Methods
The measurements of biomarkers of oxidative damage to proteins and DNA as well as lipids using liquid chromatography linked to mass spectroscopy (LCMS) will be used to establish the role of antioxidants in protecting against oxidative damage. One compound of particular interest with respect to oxidative damage to DNA is 8-hydroxydeoxyguanosine (8-OHdG). Determination of the concentration of urinary 8-OHdG will be accomplished by reverse phase HPLC with tandem mass spectrometric detection. This method requires comparatively little sample clean up while offering both measurement and confirmatory identification of 8-OHdG. 5-(hydroxymethyl)furfural (HMF) is formed as a product in the nonenzymatic browning reactions that result when food containing reducing sugars and amino acids are heated. HMF is also present in many foods including milk, fruit juice and tomato products. An LCMS base approach will be used to evaluate the mutagenicity of HMF and other metabolites derived from sugars. After standards of the proposed target HMF adducts are chemically synthesized, these synthetic standards will be fully characterized by nuclear magnetic resonance (NMR) spectroscopy, ultraviolet (UV) spectroscopy, high pressure liquid chromatography (HPLC) retention time and mass spectral (MS) fragmentation patterns. The standards will then be used for comparison with biological isolates to determine if the proposed adducts actually form when DNA is exposed to HMF. There is scant information concerning the potentially important role of clays in promoting antibiotic removal in soil systems. Knowledge of the chemical factors controlling these clay-facilitated removal and degradation pathways will aid in predicting the mobility and fate of antibiotics in soil environments. LCMS will be used to quantify the removal of two common veterinary and human antibiotics with similar chemical structures, erythromycin and clarithromycin, by clay minerals. The LC system and size exclusion chromatography with fluorescence detection will be used on organic matter extracted from local agricultural soils in order to ascertain any relevant relationships between the observed organic matter fluorescence properties and the organic matter source material. These studies will lead to more detailed mechanistic studies of the photochemical degradation of selected agrochemicals, using the LCMS system to identify and quantify the observed photoproducts. These studies will allow us to identify predictive features of the organic matter fluorescence spectra that can be used to improve the accuracy of models that are used to predict the fate and transport of agrochemicals applied to the soil surface.

Progress 07/15/07 to 07/14/08

Outputs
OUTPUTS: The high performance liquid chromatography system with mass spectral capabilities was purchased and installation completed the last week of September 2008. Training on the use of this equipment was scheduled with Shimadzu for several days in October. All the PI's completed the training as did several other members of the Chemistry and Geology Department. The PI's then spent several weeks familiarizing themselves with the instrument. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Vorlicek and Groh have utilized the LC-MS to assist in identifying novel metal-fluoroquinolone complexes. Currently, research is focusing on reactions between cadmium, p-nitrophenol, and ciprofloxacin. Results from this work are preliminary, however LC-MS and NMR spectra point to a highly stable ternary complex involving all three species. For the foreseeable future, LC-MS will continue to be used to identify new metal-fluoroquinolone-nitroaromatic compounds. This work is important because it may shed light on a currently unrecognized fate and transport pathway available to metals, antibiotics, and herbicidal degradation products within soils and natural waters. Quirk Dorr focused her use of the LC-MS on characterizing the cross-links that form between 2'-deoxyguanosine of DNA exposed to 5-[5-(5-formyl-2-thienyl)-2-furyl]-2-thiophenecarbaldehyde (NSC 629035), the proposed active metabolite 2,5-bis (5-hydroxymethyl-2-thienyl) furan (NSC 652287). NSC 652287 is derived from a family of thiophenes that are found in plants and have been used in medicine for therapeutic uses such as antibiotics, antifungal and antiviral compounds. Many of the details about how NSC 652287 reacts with DNA are unknown. NSC 629035 was examined for its activity toward 2'-deoxyguanosine. Reactions with calf-thymus DNA and sequence specific oligonucleotides were also explored. Samples were analyzed for their reactivity by chromatography methods including LC/MS. Analysis of LC/MS data has shown that NSC 652287 reacts with the calf-thymus DNA. This work has been reported by M. Thompson and D. Quirk Dorr at the American Chemical Society meeting in August 2008 entitled"Discovering the structural interactions between potential anticancer agent NSC 652287 metabolite and 2'-deoxyguanosine" (final paper number: 224) Hadley and Groh are using the LC-MS to try to identify products of reaction of malondialdehyde with folate. Initial results suggest more than one major product of the reaction but further purification of the reaction products is required. Groh is also looking at oxygen deprived combustion product analysis that has implications in the "Whoosh Bottle" demonstration done at many institutions as well as for fires on transportation vehicles carrying solvents such as alcohols. Numerous aromatic and polyaromatic hydrocarbons and have been identified and he is currently looking at the non-volatile Polycyclic aromatic hydrocarbon fraction.

Publications

  • No publications reported this period