在线净化液相色谱-串联质谱法快速测定牛奶中的6种孕激素  

Determination of six progesterones in milk byon-line cleanup liquid chromatography-tandem mass spectrometry

作　　者：段永生[1] 王炳玲[1] 艾连峰[1] 郭春海[1] 葛世辉[1] 张婧雯[1] 徐牛生 

机构地区：[1]河北出入境检验检疫局,河北石家庄050051 [2]赛默飞世尔科技(中国)有限公司,上海201206

出　　处：《色谱》2014年第6期647-652,共6页Chinese Journal of Chromatography

基　　金：国家质检总局科技计划项目(2013IK154;2014IK092)

摘　　要：建立了在线净化液相色谱-串联质谱同时测定牛奶中炔诺酮、17α-羟基孕酮、甲羟孕酮、乙酸甲地孕酮、孕酮和醋酸美伦孕酮6种孕激素的方法。本方法采用乙腈为提取溶剂提取目标化合物。提取液经在线净化柱 Cyclone-P净化，经 Phenyl-Hexyl 色谱柱分离，流动相采用0.5％（v / v）甲酸水溶液-乙腈，在电喷雾正离子模式下以多反应监测（MRM）方式测定，内标法定量。方法在0.1~50μg / L 范围内呈线性关系，线性相关系数均大于0.999。6种分析物的测定低限为0.5μg / kg，在牛奶中3个水平的添加回收率在90.8％~107.5％之间，相对标准偏差在6.3％~11.8％之间。该方法快速简便，灵敏度高，选择性好，可用于牛奶样品中孕激素的快速定性定量分析。A new method using TurboFlow on-line cleanup liquid chromatography combined with tandem mass spectrometry（MS / MS）has been developed for simultaneous determination of six progesterones including 19-norethindrone,17α-hydroxyprogesterone,medroxyprogester-one,megestrol acetate,progesterone and melengestrol acetate in milk. Samples were extracted by acetonitrile. The analytes in extract were on-line purified directly on Cyclone-P purification column where the sample matrices were washed away. Subsequently,the analytes which were eluted from the extraction column onto Phenyl-Hexyl column were separated with a gradient elution,and detected with electrospray ionization mass spectrometry in the positive scan mode using multiple reaction monitoring（ MRM）. The isotope internal standards were employed for quantification. As a result,the linearities were satisfactory with the correlation coefficients of﹥0. 999 at concentrations ranging from 0. 1 μg / L to 50 μg / L. Based on the repeated analysis of a known blank sample,the limit of quantification（LOQ）is 0. 5 μg / kg. Average recoveries of the analytes fortified at three levels（1,5 and 25 μg / kg）ranged from 90. 8% to 107. 5% ,and the relative standard deviations（RSDs）ranged from 6. 3% to 11. 8% . This proposed method is sim-ple,rapid,sensitive and highly selective,and can be applied to simultaneous identification and quantification of the six progesterones in milk.

关 键 词：在线净化 液相色谱-串联质谱 孕激素 牛奶 

分 类 号：O658[理学—分析化学]