固相萃取-LC/MS测定人血浆中腺苷蛋氨酸浓度

Determination and pharmacokinetics of S-adenosylmethionine in human plasma by solid-phase extraction-LC-MS

目的:建立固相萃取结合HPLC-MS测定人血浆中腺苷蛋氨酸(SAMe)浓度的方法,测定健康受试者单剂量口服腺苷蛋氨酸片后的血药浓度,估算其药代动力学参数。方法:血浆中加入阿昔洛韦作为内标,经高氯酸沉淀蛋白后取上清液利用固相萃取小柱进行预处理,洗脱液用HPLC-MS测定。色谱柱:Phenomenex SynerigiTM Hydro-RP C18柱(150 mm×2.0 mm,4μm),流动相:1%冰醋酸-甲醇,梯度洗脱,流速:0.2 mL/min;ESI-MS选择性正离子检测,监测离子:[M]+m/z399.2(SAMe)和[M+H]+m/z226.2(阿昔洛韦)。结果:检测方法的线性范围:0.05～5μg/mL,最低检测限:0.05μg/mL。应用此法测定了10名健康受试者单剂量口服1 000 mg腺苷蛋氨酸片的药物代谢过程。结论:本方法适合腺苷蛋氨酸的临床药代动力学研究。

Aim： To establish a rapid and sensitive solid-phase extraction - LC-MS method for the analysis of S- adenosylmethionine （SAMe） in human plasma and study its pharmacokinetics on Chinese healthy volunteers after single oral administration. Methods： Using aeielovir as the internal standard （IS）, the sample preparation involved solid-phase extraction after being deproteinized by perchloric acid. After the clean-up, samples were injected onto a Phenomenex SynerigiTM Hydro-RP C18 column （ 150 mm × 2. 0 mm, 4 μm）. The mobile phase consisted of 1% acetic acid and methanol and ran at a flow rate of 0.2 mL/min. A monopole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the[ M]^＋ ion at 399. 2 m/z for SAMe and[ M ＋ H]^＋ ion at 226.2 m/z for the internal standard. Results： The assay method was validated and linear calibration was demonstrated in the concentration range of 0. 05-5 μg/mL for SAMe with the LOD of 0. 05μg/mL. Pharmacokinetic parameters were measured in 10 healthy Chinese volunteers who receiving a single dosage at 1 000 mg of SAMe enteric-coated tablets. Conclusion： A sensitive and specific method for quantifying SAMe in human plasma has been developed and successfully applied to the clinical pharmacokinetic study of S- adenosylmethionine.