应用UE-SPE-HPLC/FLD法检测养殖业畜禽粪便中雌激素  

Estrogens determination of livestock dung based on UE-SPE-HPLC /FLD

作　　者：付银杰[1,2] 凌婉婷[1] 董长勋[2] 刘娟[1] 高彦征[1] 潘玉兰[1] 

机构地区：[1]南京农业大学土壤有机污染控制与修复研究所,南京210095 [2]南京农业大学理学院,南京210095

出　　处：《应用生态学报》2013年第11期3280-3288,共9页Chinese Journal of Applied Ecology

基　　金：江苏省科技支撑计划项目(BE2011780);国家自然科学基金项目(51278252;41171193)资助

摘　　要：本研究建立了超声提取（UE）．固相萃取净化（SPE）．高效液相色谱法（HPLC／FLD）测定畜禽粪便中雌激素（雌三醇、17β-雌二醇、炔雌醇和双酚A）的分析方法．粪便样品用乙酸乙酯超声提取30min，C18固相萃取柱净化，经Inertsil ODS—SP．C，R（150mm×4．6mm，5μm）反相色谱柱分离4种雌激素，高效液相色谱／荧光检测器（HPLC／FLD）检测，流动相采用甲醇／乙腈／水（体积比为20：30：50），激发／发射波长为280／310nm，流速0．8mL·min-1，柱温40℃，进样量20μL-结果表明：4种雌激素在1．00～1000．00μg·L-1范围内线性良好（相关系数均大于0．9995），粪便中雌三醇、双酚A、17β-雌二醇和炔雌醇的检出限分别为3．35、5．01、2．13和1．12g·kg-1；粪便样品在雌激素浓度为0．05、0．40、1．00μg·kg-1的添加水平下，猪粪、牛粪和鸡粪中4种物质的平均回收率分别为75．1％～91．1％、78．4％～117．0％和78．6％-97．8％；各物质的相对标准偏差RSD（n=6）均小于6％．采用此方法检测了南京市部分规模化养殖场的猪粪、牛粪和鸡粪样品，结果显示，利用本方法得到4种雌激素检测平行性好、检出限低，适用于畜禽粪便中雌激素的检测分析．A method for detecting the estrogens estriol, 17β-estradiol, ethinyl estradiol, and bis- phenol A in livestock dung was established by the combination of ultrasonic extraction （ UE）, solid phase extraction （SPE） purification, and high performance liquid chromatography （HPLC） with fluorescence detector （FLD）. The dung samples were extracted with ethyl acetate uhrasonication for 30 min, and purified with C18 solid phase extraction column and related solvents. The test four estrogens in the dung samples were isolated with Inertsil ODS-SP-C18 reversed-phase columns （150 min×4.6 mm, 5 μm）, and the isolated estrogens were detected with HPLC/FLD. The mo- bile phase of HPLC for the detection was methanol/acetonitrile/water （volume ratio of 20：30：50）, with a flow rate of 0.8 mL , min-1 The excitation and emission wavelengths of FLD were 280 and 310 nm, respectively, the HPLC column temperature was 40 ℃, and the injection volume was 20 μL. Good linearity （ correlation coefficient greater than 0.9995） was observed by the HPLC/FLD detection when the test four estrogens concentrations were in the range of 1.00-1000.00 μg . L-1. The detection limit of estriol, bisphenol A, 17β-estradiol, and ethinyl estradiol was 3.35, 5.01, 2.13, and 1.12 μg . kg-1, respectively. When the added estrogens concentrations of pig, cow, and chicken dung samples were 0.05, 0.40, and, 1.00 μg. kg-1 , the average recovery of the four estrogens was 75.1% -91. 1%, 78.4% -117.0%, relatively standard deviations （ RSD, n = 6 ） all less and 78.6% -97.8%, respectively, with the than 6%. By adopting the established SPE-HPLC/FLD method to detect the estrogens in real pig of the large-scale livestock raising farms in Nanjing of high, and the detection limit was low, being available in livestock dung. , cow, and chicken dung samples from parts East China, the detection reproducibility was and effective for the detection of the estrogens

关 键 词：畜禽粪便 雌激素 超声提取 固相萃取 高效液相色谱仪 荧光检测器 

分 类 号：X830.2[环境科学与工程—环境工程]