The instrument requested in this proposal is a Liquid Chromatography- Mass Spectrometry (LC-MS) system, which will be used for the analysis of toxins and other chemicals from fresh crops, such as potato and grape. The use of powerful mass spectrometric detectors in combination with liquid chromatography has played a vital role to solve many problems related to food safety. Since this technique is especially well suited for, but not restricted to the analysis of food contaminants within the food safety area, we believe that it has potential to be be utilized in many research projects in our institution. Additionally, it can be used to characterize mycotoxins in other crops, including peas, wheat and barley. <P>Our main use will be focused on analysis of toxins from fresh food crops. Acquisition of this instrumentation will improve our ability to conduct original research on the analysis of toxins from various plant sources, and therefore should improve our capacity to compete for other funding opportunities. The instrumentation and data generated also will be incorporated into the education and training of students.
The equipment sought is a Liquid Chromatography-Mass Spectrometry (LC-MS system), which is analytical chemistry technique that combines the physical separation capabilities of liquid chromatography (or HPLC) with the mass analysis capabilities of mass spectrometry. This equipment is needed for our research at North Dakota State University (NDSU) in Plant Sciences Department to have a better understanding of toxin deposition and analysis in fresh food crops. Currently, LC-MS is not available in our department.
Ochratoxin A (OTA) is generally analyzed in plants by LC, using solid phase extraction (SPE) sample preparation by using immunoaffinity column (IAC) . Analyses will be also performed by acidic methanolic extraction of samples and clean-up of extracts by IAC. The quantitative detection of toxin will be achieved by performing reverse-phase LC-fluorescence analysis. However, fluorescence detection is not complete for OTA determination, and the presence of toxin is typically confirmed by synthesis of methyl ester, or by OTA enzymatic cleavage to produce ochratoxin a (7-carboxy-5-chloro-3,4-dihydro-8-hydroxy- 3-methylisocoumarin) (30). Analysis of ochratoxin A (OTA) in grape and wine by LC-MS allows us to confirm presence of the toxin without the use of exclusive immunoaffinity columns, or time and solvent consuming sample derivatization procedures.
2012/01 TO 2012/12<br/>
OUTPUTS: The equipment we obtained from this project is a Liquid Chromatography-Mass Spectrometry (LC-MS system), which is used in the analytical chemistry technique that combines the physical separation capabilities of liquid chromatography (or HPLC) with the mass analysis capabilities of mass spectrometry. This equipment was needed for our research at North Dakota State University (NDSU) in Plant Sciences Department to have a better understanding of toxin deposition and analysis in fresh food crops. The equipment is being used by researchers at NDSU to investigate mycotoxins and glycoalkaloids in plants. One of the major fresh food crops in the USA is the potato. Glycoalkaloids have been identified as imparting resistance to some pathogen and insect pests, including early blight and Colorado potato beetle. Tuber glycoalkaloid levels are influenced by cultivar, climatic conditions, storage environment, maturity, damage during harvest handling, transport and storage, temperature exposure, and exposure to light. Little work has been directed at investigating the inheritance of glycoalkaloid levels. Parental material needs to be assessed within potato breeding programs, particularly when wild species are used in breeding efforts. Currently, one of the MS students has been quantifying these compounds in several potato genotypes grown in multiple locations. Fusarium head blight (FHB) is a major fungal disease affecting several gramineous hosts, including wheat (Triticum aestivum L.). Hard Red Spring (HRS) wheat of the Northern Great Plains of the United States and Western Provinces of Canada are susceptible to scab, especially in years that have wetter than average growing seasons. DON is the most common mycotoxin produced by Fusarium. Plants are able to "detoxify" mycotoxins such as DON by chemically modifying and/or including them in the plant matrix. These modified versions of the toxins are called "bound mycotoxins", also known as masked deoxynivalenol. One of the most common forms of bound DON is DON- 3- glucoside. In this form, a glucose molecule has been attached to the DON molecule at carbon 3. Recent studies have shown that masked DON in wheat is a cause for concern, and escapes detection by routine analytical methods. The evidence that suggests masked DON may be released into the free form under some food processing conditions, through enzymolysis in dough processing or in digestion, raises concerns that the potential toxicity of samples is being under estimated.
<br/>PARTICIPANTS: Senay Simsek, Principal Investigator Kristin Whitney, Research Specialist.
<br/>TARGET AUDIENCES: Plant scientists, Cereal Scientists, Food Scientists, potato producers, fresh crop producers,wheat and barley producers, wheat and barley end-users, food industry representatives, researchers in cereal and food sciences area, students.
<br/>PROJECT MODIFICATIONS: Not relevant to this project.
IMPACT: The utilization of advanced LC-MS technologies to food contaminants and residues has attained the determination of a broader range of compounds with higher sensitivity, selectivity and specificity. Therefore, the application of LC-MS permitted more comprehensive assessment of food safety with the determination of food contaminants and residues at trace level. Characterization of emerging food contaminants with demonstrated detrimental effects on human health is a major topic in modern Food Safety fully supported by the application of LC-MS. One of the most important trends is to develop generic methods able to extract as many contaminants (such as several mycotoxins) as possible and to detect all of them simultaneously. In one of the studies from this project we analyzed occurrence of DON and its conjugates in wheat. There is a very limited research on analysis of masked DON in HRS wheat in our region. The proposed research is the first of its kind in linking the masked DON level in FHB-infected HRS wheat and the quality defects in wheat kernels and end-products. As such, the research results will be helpful in quantifying the extent of FHB damage to wheat quality and hence its marketability. Thus, our research results will be directly helpful to wheat growers and wheat processors in both understanding the actual impact of FHB in terms of the severity of the damage and its direct economic consequence. Furthermore, state and federal regulatory agencies will be able to use the research results in establishing guidelines for both domestic and international marketing of the HRS wheat. According to the Council for Agricultural Science and Technology the annual cost to the United States because of the DON corruption of food crops was $637 million in 2003. Also, direct losses to wheat producers in United States owing to Fusarium Head Blight is estimated to be $260 million a year and total economic losses for all small grains in period of 1998-2000 is $ 2.7 billion. One of the major fresh food crops in the USA is the potato. Cultivated potato, Solanum tuberosum L., is a highly heterozygous tetraploid (4x = 48). Potatoes are adapted to an array of climates and are utilized in many ways. There are cultivars developed for different environments and end uses. Because of the wide adaptation and many species (about 1000 different Solanums, of which nearly 200 are tuber bearing), a vast genetic resource can be called upon for introgression of resistance to disease and insect pests, environmental stresses, as well as for improvement of yield and quality related traits. Research on occurrence of glycolakaloids in potato will have significant impact on potato breeding program to develop new varieties with enhanced disease package.