RPLC—ESI／MS法测定人血浆中阿普唑仑及其代谢产物的浓度

目的建立RPLC—ESI／MS法测定人血浆中阿普唑仑及其代谢产物浓度的方法。方法以地西泮为内标物；色谱柱：Agilent Zorbax SB C18柱（2．1mm×30mm，3．5μm），流动相为10mmol／L乙酰胺-乙腈（2：8），流速为0．2ml／min，柱温30℃，进样量10μl；质谱条件为电喷雾电离源（ESI），检测方式为正离子电离、多离子反应监测（MRM）。结果阿普唑仑、a-羟基阿普唑仑和4-羟基阿普唑仑在0．5～50ng／ml血浆浓度范围内线性关系良好（r=0．9996，r=0．9997，r=0．9994；n=7）；批内和批间精密度均低于5％；最低检测限为0．5ng／ml。结论该方法灵敏、准确可用于临床上阿普唑仑及a-羟基阿普唑仑和4-羟基阿普唑仑的药代动力学研究。

Determination of alprazolam in human plasma by RPLC - ESI/MS

Objective To establish an accurate and sensitive liquid chromatography - mass spectrometry （RPLC - ESI/MS） method for the determination of alprazolam and its metabolite in plasma. Methods Diazepam was used as an interal standard; The Agilent Zorbax SB C 18 （ 2.1 mm× 30 ram, 3.5μm） column was used at 30℃, with a mobile phase of 10mmol/L acetamide-acetonitrile solution（2： 8） at a flow-rate of 0.2 ml/min and injection volume of 10 μ1. Eleetrospray ionization（ESI）and multiple reaction monitoring （MRM） was applied in MS test. Results The linear calibration curves were obtained in the range of 0.5- 50 ng/ml for alprazolam,a - Hydroxyalprazolam and 4 - Hydroxyalprazolam in human plasma （ r = 0. 9996, r = 0. 9997, r = 0. 9994; n = 7） ; The limit of quantitation was 0.5 ng/ml;The intra - assay and inter - assay precision of variation was less than 5 %. Conclusion The analytical method is accurate and sensitive , it is convenient for the determination of alprazolam and its primary metabolite （a- Hydroxyalprazolam and 4- Hydroxyalprazolam） in plasma and suitable for clinical pharmacokinetic study of alprazolam.