蛋白沉淀-柱前衍生-高效液相色谱法测定畜禽血制品中甲醛

Determination of Formaldehyde in Livestock and Poultry Blood Products Using Protein Precipitation Precolumn Derivation High Performance Liquid Chromatography

目的建立畜禽血制品中甲醛含量的蛋白沉淀-柱前衍生-高效液相色谱检测方法。方法样品切碎匀浆,加入亚铁氰化钾和乙酸锌沉淀蛋白,超纯水提取,提取液加入2,4-二硝基苯肼将甲醛衍生为2,4-二硝基苯腙,再用二氯甲烷萃取水相,无水硫酸钠除水,氮吹吹干后甲醇定容,经C18色谱柱分离,紫外检测器检测与标准甲醛衍生物的保留时间对比定性,外标法定量。结果方法在2.0 mg/L40.0 mg/L线性关系良好,相关系数r=0.9991,检出限为0.2 mg/kg;加标回收率在86.0%110%之间,RSD在2.3%5.3%之间。结论样品经沉淀蛋白后,提取操作简单、效率高,衍生物性质稳定,测定灵敏度高,精密度好,适用于畜禽血制品中甲醛的快速准确检测。

Objectives To develop a method for the determination of formaldehyde in livestock and poultry blood products using protein precipitation precolumn derivation high performance liquid chromatography. Methods Samples were chopped to prepare homogenate,and potassium ferrocyanide and zinc acetate were added to precipitate the protein. Ultrapure water was adopted for extraction,2,4-dinitrophenylhydrazine was added into the extract to derive formaldehyde into 2,4-dinitrophenylhydrazone. Then,the water phase was extracted using dichloromethane,remove water using anhydrous sodium sulfate. After drying using nitrogen,methanol was used to adjust the extract to a constant volume. After separation using C18 column,the retention time of UV detector determination and the standard formaldehyde derivative were compared for qualification,and external standard method was adopted for quantification.Results The linear relationship of the method was satisfactory at 2. 0 mg/L-40. 0 mg/L with the correlation coefficient r = 0. 999 1. The detection limit was 0. 2 mg/kg,the recovery of standard addition was 86. 0%-110%,and RSD was 2. 3%-5. 3%. Conclusions After protein precipitation,the extraction operation of the sample is simple and efficient,and the property of derivatives is stable. The method has high sensitivity and precision. It is suitable for the rapid and accurate detection of formaldehyde in livestock and poultry blood products.