Optimization of Analytical Methods for Ochratoxin A and Zearalenone by UHPLC in Rice Straw Silage and Winter Forage Crops
2016
Ham, H., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Mun, H.Y., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Lee, K.A., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Lee, S., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Hong, S.K., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Lee, T., National Institute of Agricultural Sciences, Wanju, Republic of Korea | Ryu, J.G., National Institute of Agricultural Sciences, Wanju, Republic of Korea
The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with 50 μg/kg, 250 μg/kg, or 500 μg/kg of OTA and 300 μg/kg, 1500 μg/kg, or 3000 μg/kg of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity (R2 greater than equal to 0.9999 for OTA and R2 greater than equal to 0.9995 for ZEA). The limit of detection and quantification were 0.1 μg/kg and 0.3 μg/kg, respectively, for OTA and 5 μg/kg and 16.7 μg/kg, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~ 96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.
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