Determination of prostatic androgens in 10 mg of tissue using liquid chromatography-tandem mass spectrometry with charged derivatization
Determination of prostatic androgens in 10 mg of tissue using liquid chromatography-tandem mass spectrometry with charged derivatization
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DOI：
10.1007/s00216-005-3233-1
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发表时间：
2005-06-01
影响因子：
4.3
通讯作者：
Namiki, M
中科院分区：
文献类型：
--
作者：
Higashi, T;Yamauchi, A;Namiki, M
建立了测定前列腺液中5 α-双氢睾酮（DHT）和睾酮（T）的液相色谱-串联质谱（LC-MS-MS）分析方法。用MeOH-H2O（3：7，v/v）提取前列腺雄激素，用Oasis HLB柱纯化，用永久带电试剂2-肼基-1-甲基吡啶（HMP）衍生，并使用以正离子模式操作的电喷雾电离（ESI）进行LC-MS-MS分析。用HMP衍生化是非常有效的，在增加检测能力，使用积极的ESI-MS。该方法允许重复性和准确的定量ng g（-1）组织水平的前列腺雄激素在10毫克的组织。也就是说，批内和批间变异系数分别低于8.1%和9.3%，雄激素的分析回收率是定量的。DHT和T的定量限均为1.0 ng g（-1）组织。方法用于前列腺增生症和前列腺癌患者前列腺组织中DHT和T的测定，结果满意。
A practical liquid chromatography-tandem mass spectrometric (LC-MS-MS) method for the determination of prostatic 5ot-dihydrotestosterone (DHT) and testosterone (T) has been developed. The prostatic androgens were extracted with MeOH-H20 (3: 7, v/v), purified with an Oasis HLB cartridge, derivatized with the permanently charged reagent 2-hydrazino-l-methylpyridine (HMP), and subjected to LC-MS-MS analysis using electrospray ionization (ESI) operated in the positive ion mode. The derivatization with HMP was very effective at increasing the detectability using the positive-ESI-MS. The method allowed the reproducible and accurate quantification of ng g(-1) tissue levels of prostatic androgens in 10 mg of tissue. That is, the intra-and inter-assay coefficients of variation were below 8.1 and 9.3%, respectively, and the analytical recoveries of the androgens were quantitative. The limits of quantitation for DHT and T were both 1.0 ng g(-1) tissue. The developed method was used to determine DHT and T in the prostates of patients with benign prostatic hyperplasia and prostate cancer, and satisfactory results were obtained.