Evaluación de la concentración de Deoxinivalenol (DON) por cromatografía líquida de alta resolución en la población de líneas mejoradas de trigo ( Triticum aestivum)

Abstract

The Evaluation of Deoxynivalenol (DON) Concentration by High Performance Liquid Chromatography in a Population of Improved Wheat (Triticum aestivum) Lines". High Performance Liquid Chromatography in a Population of Improved Wheat (Triticum aestivum) Lines", was carried out at the was carried out at the Food Research and Analysis Services Laboratory of the Santa Catalina Experiment Station of the Santa Catalina Experimental Station of INIAP - Quito, with the objectives of validating the method for DON method of analysis of DON in wheat using Immunoaffinity columns and Liquid Chromatography of High Performance Liquid Chromatography with UV detector, and the analysis of DON content in a population of improved DON content in a population of improved lines of wheat (Triticum aestivum), corresponding to the project "Association mapping for detecting Association mapping for detecting QTLs for Fusarium Head Blight and Yellow Rust resistance in Bread Wheat Rust resistance in Bread Wheat". The methodology used was based on the R.BIOPHARM method - Ref. No.: A1-P50. for the analysis of Deoxynivalenol in wheat, for which the extraction of the analyte was carried out with 25 g of sample in 200 mL. 25 g of sample in 200 mL of bi-distilled water, 4 mL of the extract was placed in the DONPREPRE immunoaffinity column. DONPREP immunoaffinity column, it was washed with 10 mL of bi-distilled water and eluted with 3 mL of Acetonitrile grade H of HPLC-grade acetonitrile, then brought to dryness with nitrogen at 60°C and reconstituted with 300 µL of reconstituted with 300 µL of mobile phase (Water:Acetonitrile; 90:10; v/v), finally the analysis was carried out by injecting 20 µL analysis was performed by injecting 20 µL into the HPLC with UV detector (218 nm). The results of the validation showed a peak purity percentage of 100%; it is linear in a range of 250 to 2500 ng/g, obtaining a correlation coefficient r=0.9999; it is accurate since the repeatability is precise since in repeatability a RSDr ≤ 20% and reproducibility a RSDR ≤ 40% were obtained. ≤ 40%; and finally, it is accurate because a recovery percentage of 92.4% is obtained, and an uncertainty of ±40.54 ppb is associated with the final result, thus proving the validity of the analytical method. the validity of the analytical method is thus verified. This validated method was applied in the determination of DON in 327 samples of improved lines of wheat that were developed for the developed by CIMMYT and grown at INIAP's EESC to evaluate the susceptibility of wheat to DON. INIAP to evaluate susceptibility or resistance to Fusarium. A percentage of 0% incidence of DON was obtained. It is recommended that a study be conducted on the DON content in other wheat-derived products, as well as in other wheat-derived products. of DON in other wheat products, as well as in other items of interest in the national food market. the national food market.