高效液相色谱质谱联用法测定人血浆和尿液中吡西卡尼的浓度

Determination of Pilsicainide in Human Plasma and Urine by LC-MS /MS

目的建立测定人血浆和尿液中吡西卡尼的高效液相色谱质谱联用方法。方法以液液萃取法进行样品处理，以乙腈-乙酸铵（0．1‰甲酸）．水为流动相（血药浓度测定：25：37．5：37．5；尿液中药物浓度测定：35：6：59），等度洗脱，经WelchMaterialUltimateAQ—C18（2．1mm×100mm，3．5¨m）色谱柱分离，流速0．35mL·min-1、柱温25℃、进样量5灿。电喷雾离子源，正离子模式，多反应监测扫描（MRM），用于定量分析的离子反应分别为m／z273．4—110．2（吡西卡尼）和m／z287．4—110．2（内标，甲基吡西卡尼）。结果本法测定吡西卡尼浓度的线性范围分别为1～1200μg·L“（血浆），1～150mg·L-1（尿液），最低定量限分别为1μg·L-1（血浆），1mg·L-1（尿液），相关系数r值在0．9962～0．9991之间，提取回收率，基质效应，批内、批间精密度均满足药物浓度测定要求。临床药动学研究中，应用此法测定了人血浆和尿液吡西卡尼的浓度。结论本方法简便、准确、灵敏度高、重现性好，可以满足药动学研究中药物浓度测定的要求。

OBJECTIVE To develop a sensitive and specific LC-MS/MS method for the determination of pilsicainide in hu- man plasma and urine. METHODS The samples were processed by LLE（ liquid-liquid extraction） , acetonitrile-ammonium ac- etate（0.1%o formic acid） -water was used as the mobile phase with isocratie elution（25： 37.5：37.5 in plasma, 35： 6：59 in u- rine） , and separation was carried out on Welch Material Ultimate AQ-C18 column（2.1 00 × 100mm, 3.5 μm） , setting the flow rate as 0.35 mL·min-1 , the column temperature at 25℃. The injection volume was 5 txL. ESI under positive ion mode and multiple reaction monitoring scan（ MRM ） were used for quantitative analysis with m/z 273.4--＋110.2 for pilsicainide and m z 287.4--＋110.2 for internal standard, methyl-pilsicainide. RESULTS The linear ranges of the calibration curves for pilsieain- ide were 1 - 1 200 μg · L-I for plasma and 1 - 150 mg· L-1 for urine. The limitation of detection were 1 μg · L-I for plasma and 1 nag · L-1 for urine. The correlation coefficient was between 0. 996 2 and 0. 999 1. The extraction recoveries, matrix effects, and intra-/ inter-assay precisions all met the requirements of pharmacokinetic studies. This approach was applied to de- termine pilsicainide concentrations in human plasma and urine in a clinical pharmaeokinetie research. CONCLUSION This approach possesses convenience, accuracy, high sensitivity and good reproducibility, which can meet the requirements of phar- macokinetic studies.