This study aims to establish an analytic approach for determining isochlorogenic acid content in feed using high performance liquid chromatography(HPLC).Related parameters are as follows:C18 chromatographic column(250 mm × 4.6 mm×5 μm),mobile phases(A:acetonitrile∶water∶formic acid at 250∶750∶1;and B:acetonitrile∶water∶formic acid at 100∶900∶1),gradient elution,1.5 mL/min flow rate,25℃ column temperature,330 nm detection wavelength,and 10 μL sam-pling size.After undergoing extraction with 60％methanol solution and albumen precipitation,the sample feed was then purified through MAX mixed anion-exchange column,tested and ESTD(external standard method)quantified after passing the organic filterable membrane.The results indicate 3 isochlorogenic acids are well separated without miscellaneous peak interference and the linearity appears good within the range of 0.05-5 μg/mL.The approach in question could reduce the effect of complicated feed substrate on test results and features high accuracy and repeatability.When sampling size is 1.0 g,the limit of detection is 5.0 mg/kg for all three isochlorogenic acids,A,B and C.The study concludes the applicability of the proposed HPLC(high per-formance liquid chromatography)approach to accurate detection of isochlorogenic acids A,B and C in feed,which would provide technical support for massive application and management of isochlorogenic acids A,B and C in the feed.
维普
万方数据