Measurement of tamsulosin in human serum by liquid chromatography-tandem mass spectrometry

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• 作者：Rita Upreti ; Natalie Z.M. Homer ; Gregorio Naredo ; Diego F. Cobice ; Katherine A. Hughes ; Laurence H. Stewart ; Brian R. Walker ; Ruth Andrew
• 关键词：BPH ; benign prostatic hyperplasia
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2013
• 出版时间：1 July, 2013
• 年：2013
• 卷：930
• 期：Complete
• 页码：121-128
• 全文大小：1024 K

文摘

A simple, sensitive and robust method to extract tamsulosin from human serum, and quantify by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated and is applicable as a measure of compliance in clinical research. Tamsulosin was extracted from human serum (100 ¦ÌL) via liquid-liquid extraction with methyl tert-butyl ether (2 mL) following dilution with 0.1 M ammonium hydroxide (100 ¦ÌL), achieving 99.9 % analyte recovery. Internal standard, d9-finasteride, was synthesised in-house. Analyte and internal standard were separated on an Ascentis^? Express C18 (100 mm ¡Á 3 mm, 2.7 ¦Ìm) column using a gradient elution with mobile phases methanol and 2 mM aqueous ammonium acetate (5:95, v/v). Total run-time was 6 min. Tamsulosin was quantified using a triple quadrupole mass spectrometer operated in multi-reaction-monitoring (MRM) mode using positive electrospray ionisation. Mass transitions monitored for quantitation were: tamsulosin m/z 409 ¡ú 228 and d9-finasteride m/z 382 ¡ú 318, with the structural formulae of ions confirmed by Fourier transform ion cyclotron resonance mass spectrometry (within 10 ppm). The limit of quantitation was 0.2 ng/mL, and the method was validated in the linear range 0.2-50 ng/mL with acceptable inter- and intra-assay precision and accuracy and stability suitable for routine laboratory practice. The method was successfully applied to samples taken from research volunteers in a clinical study of benign prostatic hyperplasia.