高效液相色谱-质谱法测定比格犬血浆中的艾普拉唑及其药代动力学

运用高效液相色谱-电喷雾质谱(HPLC-ESI-MS)技术,建立了快速、简单、灵敏的比格犬静脉滴注艾普拉唑钠盐后血药浓度的检测方法。血浆样品采用蛋白沉淀法,以丁螺环酮作为内标,色谱柱为Teknokroma Kromasil C18(100 mm×2.1 mm, 5 μm),流动相为水-甲醇-乙腈(69:8:23, v/v/v)(含0.1%的甲酸),流速0.2 mL/min,采用电喷雾(ESI)离子源以正离子方式检测。绘制血药浓度-时间曲线,并采用DAS 2.0计算药代动力学参数。方法学实验结果表明内源性杂质不干扰艾普拉唑和内标的测定,线性范围为5～10000 μg/L (r=0.994),最低定量限为5 μg/L,精密度和准确度均符合生物样品测定的要求。低、中、高3个浓度的绝对回收率在106%左右,基质效应小于142.0%,表明该方法适合比格犬血浆中艾普拉唑浓度的测定及药代动力学研究。比格犬静脉滴注艾普拉唑钠盐3个剂量(0.2 mg/kg、0.8 mg/kg和3.2 mg/kg)后的药-时曲线下面积(AUC(0~∞))分别为(2.4×104±3×103)、(8.8×104±1.6×104)和(5.4×105±8×104) μg/L•min,呈线性药物代谢动力学过程。

Determination of ilaprazole in beagle plasma and its pharmaeokineties by high performance liquid chromatography-mass spectrometry

A sensitive, simple and specific high performance liquid chromatography-electro spray ionization mass spectrometry method was developed for the determination of ilaprazole in the plasma of beagles administered via i.v. bolus doses of ilaprazole. The procedure employed buspirone as the internal standard and a simple protein precipitation step. The separation was achieved using a C18 column (100 mm x 2.1 mm, 5 microm) with a mobile phase consisting of water-methanol-acetonitrile (69:8:23, v/v/v) containing 0.1% (v/v) formic acid at a flow rate of 0.2 mL/min. The detection was accomplished by a mass spectrometer using selected ion monitoring (SIM) in positive mode. The linearity was from 5 microg/L to 10,000 microg/L with a sensitivity of 5 microg/L as the lower limit of quantification. The inter- and intra- day precisions were within 9.00%. The mean recoveries at three spiked levels were about 106% and the matrix effects were less than 142.0%. The method described above was successfully applied to analyze the beagle plasma samples of ilaprazole in a pharmacokinetic study. The area under the plasma concentration-time curve (AUC(0-infinity) of ilaprazole after i.v. doses of 0.2, 0.8 and 3.2 mg/kg were (2.4 x 10(4) +/- 3 x 10(3)), (8.8 x 10(4) +/- 1.6 x 10(4)) and (5.4 x 10(5) +/- 8 x 10(4)) microg/L x min, respectively. On the basis of AUC, the pharmacokinetic property of ilaprazole was proposed to be linear dynamics.