动物营养学报
動物營養學報
동물영양학보
ACTA ZOONUTRIMENTA SINICA
2015年
6期
1823-1831
,共9页
安亚南%王丹阳%丁立人%杨新岗%邓亚军%刘强
安亞南%王丹暘%丁立人%楊新崗%鄧亞軍%劉彊
안아남%왕단양%정립인%양신강%산아군%류강
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
黃麯黴毒素B1%玉米赤黴烯酮%黴菌毒素吸附劑%豬食糜上清液%吸附效果
황곡매독소B1%옥미적매희동%매균독소흡부제%저식미상청액%흡부효과
aflatoxin B1%zearalenone%mycotoxin adsorbent%supernatant liquor of chyme of pigs%binding ef-ficacy
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1.00~10000.00 ng/mL黄曲霉毒素B1和20.00~5000.00 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。
本試驗旨在建立測定豬胃和小腸食糜上清液中黃麯黴毒素B1和玉米赤黴烯酮含量的高效液相色譜( HPLC)法,併利用該方法評價黴菌毒素吸附劑分彆在不同介質中對黃麯黴毒素B1和玉米赤黴烯酮的吸附效果。為消除食糜上清液中複雜組分對目標毒素檢測的榦擾,本試驗對毒素提取方法、液相色譜的檢測波長設置和流動相組成進行瞭優化。利用單濃度體外吸附試驗評價瞭4種黴菌毒素吸附劑分彆在水、豬胃和小腸食糜上清液介質中對黃麯黴毒素B1和玉米赤黴烯酮的吸附效果。結果錶明：1)反應上清液經過1次正己烷和3次二氯甲烷提取,熒光激髮波長和髮射波長在0~9 min和9~14 min分彆設定為360、440 nm和235、418 nm,乙腈/水流動相中乙腈比例控製在40%~60%時,本試驗採用的HPLC儀對1.00~10000.00 ng/mL黃麯黴毒素B1和20.00~5000.00 ng/mL玉米赤黴烯酮具有良好的分離定量效果,線性相關繫數分彆為1.000和0.999；檢齣限分彆為0.16和2.00 ng/mL；黃麯黴毒素B1和玉米赤黴烯酮日內精密度相對標準差分彆為2.37%、4.50%；日間精密度相對標準差分彆為2.74%、8.84%。黃麯黴毒素B1和玉米赤黴烯酮標準添加迴收率分彆為89.1%~110.3%、98.7%~104.1%。2)自製黴菌毒素吸附劑樣品P3、M3和市場採購的黴菌毒素吸附劑產品Y1、Y2在豬胃食糜上清液介質中對黃麯黴毒素B1的吸附率依次為72.10%、84.18%、83.17%和66.01%；在豬小腸食糜上清液介質中的吸附率依次為78.15%、88.08%、88.12%和67.34%。4種吸附劑在胃食糜上清液介質中對玉米赤黴烯酮的吸附率依次為85.35%、18.41%、16.20%和12.98%；在豬小腸食糜上清液介質中的吸附率分彆為39.75%、25.15%、21.40%和24.85%。綜上,本試驗建立的HPLC法可同時測定豬胃和小腸食糜上清液中的黃麯黴毒素B1和玉米赤黴烯酮,檢測限和檢測範圍滿足我國《豬飼料衛生標準》的要求。該方法可用于評價黴菌毒素吸附劑豬生理體液條件下對黃麯黴毒素B1和玉米赤黴烯酮的吸附效果。
본시험지재건립측정저위화소장식미상청액중황곡매독소B1화옥미적매희동함량적고효액상색보( HPLC)법,병이용해방법평개매균독소흡부제분별재불동개질중대황곡매독소B1화옥미적매희동적흡부효과。위소제식미상청액중복잡조분대목표독소검측적간우,본시험대독소제취방법、액상색보적검측파장설치화류동상조성진행료우화。이용단농도체외흡부시험평개료4충매균독소흡부제분별재수、저위화소장식미상청액개질중대황곡매독소B1화옥미적매희동적흡부효과。결과표명：1)반응상청액경과1차정기완화3차이록갑완제취,형광격발파장화발사파장재0~9 min화9~14 min분별설정위360、440 nm화235、418 nm,을정/수류동상중을정비례공제재40%~60%시,본시험채용적HPLC의대1.00~10000.00 ng/mL황곡매독소B1화20.00~5000.00 ng/mL옥미적매희동구유량호적분리정량효과,선성상관계수분별위1.000화0.999；검출한분별위0.16화2.00 ng/mL；황곡매독소B1화옥미적매희동일내정밀도상대표준차분별위2.37%、4.50%；일간정밀도상대표준차분별위2.74%、8.84%。황곡매독소B1화옥미적매희동표준첨가회수솔분별위89.1%~110.3%、98.7%~104.1%。2)자제매균독소흡부제양품P3、M3화시장채구적매균독소흡부제산품Y1、Y2재저위식미상청액개질중대황곡매독소B1적흡부솔의차위72.10%、84.18%、83.17%화66.01%；재저소장식미상청액개질중적흡부솔의차위78.15%、88.08%、88.12%화67.34%。4충흡부제재위식미상청액개질중대옥미적매희동적흡부솔의차위85.35%、18.41%、16.20%화12.98%；재저소장식미상청액개질중적흡부솔분별위39.75%、25.15%、21.40%화24.85%。종상,본시험건립적HPLC법가동시측정저위화소장식미상청액중적황곡매독소B1화옥미적매희동,검측한화검측범위만족아국《저사료위생표준》적요구。해방법가용우평개매균독소흡부제저생리체액조건하대황곡매독소B1화옥미적매희동적흡부효과。
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 intesti-nal 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 e-valuated 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 mo-bile 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 recov-ery 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 simultane-ous 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.
