School of Pharmacy, Guangdong Medical College, Dongguan 523808, PR China.
J Pharm Biomed Anal. 2013 Feb 23;74:56-61. doi: 10.1016/j.jpba.2012.10.021. Epub 2012 Nov 1.
A simple, sensitive and specific liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) method was developed and validated to quantitate nevadensin in rat plasma using genistein as the internal standard (IS). The assay was based on protein precipitation treatment with acetonitrile and liquid chromatography performed on a reverse-phase Zorbax SB-C(18) column (50 mm × 2.1 mm, 1.8 μm). Elution was achieved with a mobile phase consisted of aqueous solution containing 0.1% formic acid (solvent A) and methanol solution containing 0.1% formic acid (solvent B) at a flow rate of 300 μL/min (30:70, v/v). Quantification was through tandem-mass spectrometry with positive electrospray ionization (ESI) and multiple reaction monitoring (MRM) at m/z 345.1→315.1 and 271.1→215.1 for nevadensin and IS, respectively. Calibration curves were linear over the nevadensin rat plasma concentration range of 0.1-300 ng/mL. The lower limit of quantification (LLOQ) was 0.100 ng/mL and the inter- and intra-day accuracy and precision ranged between -1.67% and 7.60%. The average recovery of nevadensin was 95.6-99.1%. The validated method was successfully applied to the pharmacokinetic evaluation of nevadensin using the rat as an animal model following an oral administration of 50mg/kg nevadensin.
建立并验证了一种简单、灵敏、特异的液相色谱-电喷雾串联质谱（LC-MS/MS）法，用于测定大鼠血浆中的新穿心莲内酯。以染料木黄酮为内标（IS），采用乙腈沉淀蛋白处理，通过反相 Zorbax SB-C18 柱（50mm×2.1mm，1.8μm）进行色谱分离。流动相由含 0.1%甲酸的水溶液（溶剂 A）和含 0.1%甲酸的甲醇溶液（溶剂 B）组成，流速为 300μL/min（30:70，v/v）进行洗脱。通过正电喷雾离子化（ESI）和多反应监测（MRM）进行串联质谱检测，新穿心莲内酯和 IS 的质荷比分别为 345.1→315.1 和 271.1→215.1。新穿心莲内酯在大鼠血浆中的浓度范围为 0.1-300ng/mL 时呈线性关系。定量下限（LLOQ）为 0.100ng/mL，日内和日间准确度和精密度的范围分别为-1.67%至 7.60%。新穿心莲内酯的平均回收率为 95.6-99.1%。该方法已成功应用于大鼠口服 50mg/kg 新穿心莲内酯后的药代动力学评价。
