酶解-UPLC-MS/MS法同时测定咸鸭蛋黄中的11种胆固醇氧化物  

Simultaneous Determination of 11 Cholesterol Oxidation Products in Salted Duck Egg Yolk by Enzymolysis-Liquid Chromatography Tandem Mass Spectrometry

作　　者：柯星[1] 黄百芬[2] 吕美玲[3] 莫卫民[1] 任一平[2] 

机构地区：[1]浙江工业大学化学工程学院,杭州310014 [2]浙江省疾病预防控制中心,杭州310051 [3]安捷伦科技中国有限公司,北京100102

出　　处：《分析化学》2015年第12期1920-1926,共7页Chinese Journal of Analytical Chemistry

基　　金：浙江省公益性技术应用研究计划(No.2015C37061)资助~~

摘　　要：建立了酶解-超高效液相色谱-串联质谱法（UPLC-MS/MS）同时检测咸鸭蛋黄中11种胆固醇氧化物（COPs）的方法。咸鸭蛋黄样品经过脂肪酶37℃酶解,正己烷-乙醚混合液（8∶2,V/V）提取,Silica固相萃取柱净化。采用Waters HSS T3柱（100 mm×2.1 mm,0.3μm）,以0.1%甲酸的水相和甲醇/乙腈（1∶1,V/V）混合液的有机相梯度洗脱,实现了11种COPs的基线分离。以d7-7α-OH-胆固醇为内标,在APCI-MS/MS MRM模式下定量检测。结果表明,11种COPs在对应的20~1200 ng/m L和200~3500 ng/m L线性范围内,相关系数均大于0.996,3个加标水平的回收率在79.3%~104.1%之间,日间RSD在2.0%~12.9%之间;定量限范围在0.08~0.40μg/g之间。本方法具有较好的重复性和准确性,并成功应用于咸鸭蛋黄中的11种COPs含量的分布检测。A method for simultaneous quantification of 11 cholesterol oxidation products（ COPs） in yolk of salted duck egg was developed by ultra performance liquid chromatography-tandem mass spectrometry（ UPLCMS / MS） with enzymatic treatment. Firstly,the yolk of salted duck egg was digested by lipase at 37℃,and COPs were extracted by n-hexane and diethyl ether mixture（ 8∶ 2,V / V）. The supernatant was enriched and purified by solid phase extraction of silica. Eleven COPs were separated by UPLC HSS T3 column（ 100 mm× 2. 1 mm,0. 3 μm） with water and methanol / acetonitrile（ 1∶ 1,V / V） both containing 0. 1% formic acid.All COPs were simultaneously determined by UPLC-MS / MS with multiple reaction monitoring mode（ MRM）.The d7-7α-OH cholesterol was used as internal standard. The results showed that the linear ranges were20- 1200 ng / m L and 200- 3500 ng / m L with correlation coefficient（ r ≥0. 996）. The recoveries of three spiking levels were between 79. 3% and 104. 1%,and relative standard deviations（ RSD） were 2. 0% and12. 9%. The limit of quantification（ LOQ） was 0. 08- 0. 40 μg / g. The established method with good repeatability and robust recovery can be applied to quantify the levels of 11 COPs in yolk of salted duck eggs.

关 键 词：胆固醇氧化物 咸鸭蛋黄 酶解 液相色谱串联质谱 

分 类 号：O657.63[理学—分析化学] TS253.7[理学—化学]