Development of a multianalyte method for the determination of anabolic hormones in bovine urine by isotope-dilution GC–MS/MS
Development of a multianalyte method for the determination of anabolic hormones in bovine urine by isotope-dilution GC–MS/MS
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DOI：
10.1007/s00216-006-0739-0
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发表时间：
2006-09
影响因子：
4.3
通讯作者：
C. S. Aman;A. Pastor;G. Cighetti;M. Guardia
中科院分区：
文献类型：
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作者：
C. S. Aman;A. Pastor;G. Cighetti;M. Guardia
建立了一种灵敏、特异、选择性好的多组分GC-MS/MS法测定牛尿中11种蛋白同化激素的方法。将尿液pH值调节至4.8后，向样品中加入氘代内标物，并使用β-葡萄糖醛酸酶/芳基硫酸酯酶进行酶水解。激素用甲醇通过C18固相柱洗脱，并进行液-液萃取。分析物经N，O-双（三甲基硅基）三氟乙酰胺（BSTFA）和1%三甲基氯硅烷衍生化，GC-MS数据在正电子轰击串联质谱模式下获得。在这些条件下，未观察到基质效应，检测限值在0.005 ng/mL（己烯雌酚）至0.38 ng/mL（17α-甲基睾酮和17α-乙炔基睾酮）范围内。在选定的条件下，回收率为81%（α-玉米赤霉醇）~ 149%（17α-甲基睾酮）。这些结果优于使用七氟丁酸酐（HFBA）作为衍生试剂，并在全扫描和选择性离子监测模式下测量的结果。
A sensitive, specific and selective multianalyte GC–MS/MS method has been developed for the determination of 11 anabolic hormones in bovine urine. After adjusting the urine pH to 4.8, the samples were spiked with deuterated internal standards and submitted to enzymatic hydrolysis with β-glucuronidase/arylsulfatase. Hormones were eluted with methanol through a C18 solid phase cartridge and submitted to a liquid–liquid extraction. Analytes were derivatized by addingN,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane and GC–MS data were obtained in the positive electron impact tandem mass mode. Under these conditions, no matrix effects were observed and limit of detection values were in the range of 0.005 ng/mL (diethylstilbestrol) to 0.38 ng/mL (17α-methyltestosterone and 17α-ethynylestradiol). Recoveries from 81% (α-zeranol) to 149% (17α-methyltestosterone) were found under the selected conditions. These results were better than those found using heptafluorobutyric anhydride (HFBA) as derivative reagent and those measured in full scan and selective ion monitoring modes.