UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物

Determination of Simvastatin and Simvatatin Acid in Human Plasma by UHPLC-MS/MS

关键词:  辛伐他汀  辛伐他汀酸  液质联用  药动学

Key words:  simvastatin  simvatatin acid  UHPLC-MS/MS  pharmacokinetics

摘要:

目的 采用UHPLC-MS/MS同时测定人血浆中辛伐他汀及其代谢产物辛伐他汀酸，并研究辛伐他汀、辛伐他汀酸在人体内的药动学特征。方法 血浆样品以乙醚萃取，采用UHPLC-MS/MS进行分析。色谱柱：Agilent ZORBAX SB-C₁₈ (100 mm×2.1 mm，3.5 μm)；乙腈-1 mmol?L^-1醋酸铵(甲酸调pH 4.5)为流动相梯度洗脱，流速：0.2 mL?min^-1。采用电喷雾离子源(ESI)，以多反应监测方式(MRM)进行定量分析。辛伐他汀和内标洛伐他汀在正离子模式下定量分析，离子对分别为m/z 419.4→199.3和m/z 405.3→199.3；辛伐他汀酸和内标洛伐他汀酸在负离子模式下定量分析，离子对分别为m/z 435.5→115.2和m/z 421.4→101.2。结果 辛伐他汀和辛伐他汀酸的线性范围均为0.2~50 ng?mL^-1(r>0.99)，最低定量限均为0.2 ng?mL?1，日内和日间精密度(RSD)均≤11.10%，提取回收率均≥63.71%。结论 该方法专属性强、灵敏度高、重现性好，适用于辛伐他汀的药动学研究。

Abstract:

OBJECTIVE To develop a UHPLC-MS/MS method for the simultaneous determination of simvastatin and its metabolite simvastatin acid in human plasma. METHODS The sample was extracted with diethylether and then separated on an Agilent ZORBAX SB-C₁₈(100 mm×2.1 mm, 3.5 μm), with acetonitrile and 1 mmol?L^-1 ammonium acetate(pH was adjusted to 4.5 by formic acid) as mobile phase eluted by a gradient program at a flow rate of 0.2 mL?min^-1. Detection was performed with multiple reactions monitoring(MRM) using electrospray ionization(ESI). The positive ion scan mode was used to detect simvastatin and lovastatin(IS) while the negative ion scan mode was used to detect simvastatin acid and lovastatin acid(IS). The MRM transitions of m/z 419.4/199.3 and m/z 405.3/199.3 were used to quantify simvastatin and lovastatin while m/z 435.5/115.2 and m/z 421.4/101.2 were used to quantify simvastatin acid and lovastatin acid. RESULTS The calibration curves were linear over the concentration range of 0.2?50 ng?mL^-1(r>0.99) with the lower limit of quantitation 0.2 ng?mL^-1. Both inter- and intra-day relative standard deviations were ≤11.10%. The extraction recoveries of simvastatin and simvastatin acid were ≥63.71%. CONCLUSION The UHPLC-MS/MS method is selective, sensitive, reliable and suitable for pharmacokinetic study of simvastatin.