高效液相色谱-质谱联用法测定人血浆中美利曲辛的含量

Determination of melitracen in human plasma by HPLC-MS

目的：建立测定人血浆中美利曲辛浓度的高效液相色谱-质谱联用法。方法：血浆样品中加入内标特非那定，用醋酸乙酯萃取。色谱柱为Thermo Hypersil-HyPURITY C18（150mm×2．1mm，5μm），流动相为10mmol·L^-1醋酸铵溶液（pH3．9）-甲醇-乙腈（41：5：54），流速0．25mL·min^-1。采用电喷雾电离源正源（ESI^＋），选择离子检测方式，用于定量分析的检测离子为m/z 292（美利曲辛）和m/z 474（内标）。结果：美利曲辛在0．78-50．0μg·L^-1内线性关系良好（r=0．9998），最低定量浓度为0．78μg·L^-1。方法回收率在92．3％-01．0％之间，日内与日间RSD均小于15％（n=5）。结论：该方法灵敏度高，结果准确，适用于治疗剂量美利曲辛体内血药浓度的测定。

OBJECTIVE To develop an HPLC-MS method for the determination of melitracen in human plasma. METHODS Melitracen and the internal standard terfenadine added as internal standard were extracted from the patient plasma using liquid-liquid extraction by ethyle acetate, then separated on a Thermo Hypersil-HyPURITY C18 （150 mm ×2. 1 mm, 5 μm）with a mobile phrase of 10 mmol·L^-1 ammonium acetate -methanol-acetonitrile （41：5：54） at a flow rate of 0. 25 mL·min^-1. HPLC- MS-ESI^＋ was performed in the selected ion monitoring（SIM） mode using target ions at m/z 292 （melitracen） and m/z 474 （IS）. RESULTS The linear concentration range of the calibration curves for melitracen was 0. 78 - 50. 0μg·L^-1 （r= 0. 999 8）. The limit of quantitation of melitracen was 0. 78μg·L^-1. The average recovery was in the range of 92. 3% - 101.0%. Withinday and between-day RSDs were less than 15% （n = 5）. CONCLUSION The method is sensitive,accurate and suitable for the determination of melitracen in Human Plasma.