Quantitation of anabolic hormones and their metabolites in bovine serum and urine by liquid chromatography-tandem mass spectrometry
Quantitation of anabolic hormones and their metabolites in bovine serum and urine by liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/s0021-9673(99)01293-5
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发表时间：
2000-02-18
影响因子：
4.1
通讯作者：
Cammarata, P
中科院分区：
文献类型：
--
作者：
Draisci, R;Palleschi, L;Cammarata, P
报道了一种基于串联质谱和使用常压化学电离的在线高效液相色谱 (LC-APCI-MS-MS) 的特异性和灵敏方法，用于定量牛血清和尿液中的合成代谢激素残留物（17 β-19-去甲睾酮、17 β-睾酮和黄体酮）及其主要代谢物（17 α-19-去甲睾酮和 17 α-睾酮）。 [H-2(2)]17 β-睾酮用作内标。通过液-液萃取从尿液（酶水解后）和血清样品中提取分析物，并通过 C-18 固相萃取纯化。电离在以正离子模式运行的加热雾化器接口中进行，其中每种分析物仅生成质子化分子 [M+H](+)。它充当碰撞诱导解离的前体离子，并且通过选择的反应监测 LC-MS-MS 鉴定了每种分析物的两个诊断产物离子，以明确激素确认。牛血清和尿液样本的总体日间精密度（相对标准偏差）分别为6.37%至2.10%和6.25%至2.01%，而牛血清和尿液样本的日间准确度（相对误差）分别为-5.90%至-3.18%和-6.40%至-2.97%。该方法对牛血清和尿液中所有激素的定量限为0.1 ng/ml。由于其高灵敏度和特异性，该方法已成功用于确认出于监管目的的非法激素施用。 (C) 2000 年由 Elsevier Science B.V. 出版。保留所有权利。
A specific and sensitive method based on tandem mass spectrometry with on-line high-performance liquid chromatography using atmospheric pressure chemical ionisation (LC-APCI-MS-MS) for the quantitation of anabolic hormone residues (17 beta-19-nortestosterone, 17 beta-testosterone and progesterone) and their major metabolites (17 alpha-19-nortestosterone and 17 alpha-testosterone) in bovine serum and urine is reported. [H-2(2)]17 beta-Testosterone was used as internal standard. The analytes were extracted from urine (following enzymatic hydrolysis) and serum samples by liquid-liquid extraction and purified by C-18 solid-phase extraction. Ionisation was performed in a heated nebulizer interface operating in the positive ion mode, where only the protonated molecule, [M+H](+), was generated for each analyte. This served as precursor ion for collision-induced dissociation and two diagnostic product ions for each analyte were identified for the unambiguous hormone confirmation by selected reaction monitoring LC-MS-MS. The overall inter-day precision (relative standard deviation) ranged from 6.37 to 2.10% and from 6.25 to 2.01%, for the bovine serum and urine samples, respectively, while the inter-day accuracy (relative error) ranged from -5.90 to -3.18% and from -6.40 to -2.97%, for the bovine serum and urine samples, respectively. The limit of quantitation of the method was 0.1 ng/ml for all the hormones in bovine serum and urine. On account of its high sensitivity and specificity the method has been successfully used to confirm illegal hormone administration for regulatory purposes. (C) 2000 Published by Elsevier Science B.V. All rights reserved.