动物营养学报 >

2015 , Vol. 27 >Issue 6: 1823 - 1831

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.06.019

饲料安全 Feed Security

猪胃肠道食糜中黄曲霉毒素B1和玉米赤霉烯酮检测方法的改进及其在霉菌毒素吸附剂吸附效果评价中的应用

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  • 南京农业大学动物科技学院, 南京 210095
安亚南(1988—),女,河南开封人,硕士研究生,动物营养与饲料科学专业。E-mail: 2012105048@njau.edu.cn

收稿日期: 2015-01-23

  网络出版日期: 2015-06-17

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Simultaneous Measurement of Aflatoxin B1 and Zearalenone in Gastrointestinal Chyme of Pigs and Assessment of Binding Efficacy of Mycotoxin Adsorbents

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  • College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Received date: 2015-01-23

  Online published: 2015-06-17

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摘要

本试验旨在建立测定猪胃和小肠食糜上清液中黄曲霉毒素B1和玉米赤霉烯酮含量的高效液相色谱(HPLC)法,并利用该方法评价霉菌毒素吸附剂分别在不同介质中对黄曲霉毒素B1和玉米赤霉烯酮的吸附效果。为消除食糜上清液中复杂组分对目标毒素检测的干扰,本试验对毒素提取方法、液相色谱的检测波长设置和流动相组成进行了优化。利用单浓度体外吸附试验评价了4种霉菌毒素吸附剂分别在水、猪胃和小肠食糜上清液介质中对黄曲霉毒素B1和玉米赤霉烯酮的吸附效果。结果表明:1)反应上清液经过1次正己烷和3次二氯甲烷提取,荧光激发波长和发射波长在0~9 min和9~14 min分别设定为360、440 nm和235、418 nm,乙腈/水流动相中乙腈比例控制在40%~60%时,本试验采用的HPLC仪对1.00~10 000.00 ng/mL黄曲霉毒素B1和20.00~5 000.00 ng/mL玉米赤霉烯酮具有良好的分离定量效果,线性相关系数分别为1.000和0.999;检出限分别为0.16和2.00 ng/mL;黄曲霉毒素B1和玉米赤霉烯酮日内精密度相对标准差分别为2.37%、4.50%;日间精密度相对标准差分别为2.74%、8.84%。黄曲霉毒素B1和玉米赤霉烯酮标准添加回收率分别为89.1%~110.3%、98.7%~104.1%。2)自制霉菌毒素吸附剂样品P3、M3和市场采购的霉菌毒素吸附剂产品Y1、Y2在猪胃食糜上清液介质中对黄曲霉毒素B1的吸附率依次为72.10%、84.18%、83.17%和66.01%;在猪小肠食糜上清液介质中的吸附率依次为78.15%、88.08%、88.12%和67.34%。4种吸附剂在胃食糜上清液介质中对玉米赤霉烯酮的吸附率依次为85.35%、18.41%、16.20%和12.98%;在猪小肠食糜上清液介质中的吸附率分别为39.75%、25.15%、21.40%和24.85%。综上,本试验建立的HPLC法可同时测定猪胃和小肠食糜上清液中的黄曲霉毒素B1和玉米赤霉烯酮,检测限和检测范围满足我国《猪饲料卫生标准》的要求。该方法可用于评价霉菌毒素吸附剂猪生理体液条件下对黄曲霉毒素B1和玉米赤霉烯酮的吸附效果。

关键词： 黄曲霉毒素B1; 玉米赤霉烯酮; 霉菌毒素吸附剂; 猪食糜上清液; 吸附效果

本文引用格式

安亚南, 王丹阳, 丁立人, 杨新岗, 邓亚军, 刘强 . 猪胃肠道食糜中黄曲霉毒素B1和玉米赤霉烯酮检测方法的改进及其在霉菌毒素吸附剂吸附效果评价中的应用[J]. 动物营养学报, 2015 , 27(6) : 1823 -1831 . DOI: 10.3969/j.issn.1006-267x.2015.06.019

Abstract

This experiment was conducted to develop a high-performance liquid chromatographic (HPLC) method for quantitative determination of aflatoxin B1 and zearalenone in supernatant liquor of gastric and intestinal of pigs, and the method was used to evaluate binding efficacy of aflatoxin B1 and zearalenone by mycotoxin absorbents in different media. In order to eliminate the influence of complex composition in the supernatant, the experiment optimized extraction and purification of the supernatant, detection wavelengths and mobile phases of liquid chromatography. Binding efficacy of 4 mycotoxin adsorbents in a fixed mycotoxin concentration was evaluated respectively in purified water,the supernatant liquor of gastric and intestinal chyme of pigs. The results showed as follows: 1) the supernatant liquor samples were extracted with n-hexane and dichloromethane, the optimal chromatographic condition was that the fluorescent emitted light and excitation light wavelength were set as 360, 440 nm within 0 to 9 minutes and 235, 418 nm within 9 to 14 minutes, respectively, and the mobile phase was the mixture of acetonitrile and water with acetonitrile percentage in the range of 40% to 60%. The analytical method was quantitative determination for aflatoxin B1 from 1.00 to 10 000.00 ng/mL and zearalenone from 20.00 to 5 000.00 ng/mL, respectively. The linear correlation coefficient of aflatoxin B1 was 1; the limit of detection was 0.16 ng/mL, the intra-day and inter-day precision were 2.37% and 2.74%, with recoveries ranging from 89.1% to 100.3%. The linear correlation coefficient of zearalenone was 0.999; the limit of detection was 2.00 ng/mL, the intra-day and inter-day precision were 4.50% and 8.84%, with recovery ranging from 98.7% to 104.1%. 2) The adsorption rates of aflatoxin B1 on P3, M3, Y1 and Y2 were 72.10%, 84.18%, 83.17% and 66.01% from gastric supernatant of pigs, and 78.15%, 88.08%, 88.12% and 67.34% from intestinal supernatant,respectively. The binding rate of zearalenone on P3, M3, Y1 and Y2 were 85.35%, 18.41% and 16.20% from gastric supernatant of pigs, and 12.98%, 39.75%, 25.15%, 21.40% and 24.85% from intestinal supernatant,respectively. In conclusion, the HPLC method is developed for simultaneous measurement of aflatoxin B1 and zearalenone in supernatant liquor of gastric and intestinal chyme of pigs, and its detection limits and ranges meet the requirement of Hygienical Standard for Feeds in our country. The method is applied to evaluate binding efficacy of mycotoxin adsorbents under the condition of physiological body fluid of pigs.

Key words： aflatoxin B1; zearalenone; mycotoxin adsorbent; supernatant liquor of chyme of pigs; binding efficacy

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