人血浆中盐酸沙格雷酯的UPLC-MS/MS测定及药代动力学研究

Quantification of sarpogrelate hydrochloride in human plasma by UPLC-MS/MS and its application in a pharmacokinetic study

建立一种快速、准确测定人血浆中盐酸沙格雷酯的UPLC-MS/MS分析方法。以阿立哌唑为内标,0.2 m L含药血浆经乙腈沉淀蛋白后进样分析;流动相组成为乙腈:0.01 mol/L甲酸铵水溶液=30∶70,梯度洗脱方式,乙腈比例在2 min内从30%变为65%,流量0.3 m L/min,柱温为35℃,进样量3μL。质谱条件:气动辅助电喷雾离子化(ESI)源;正离子检测(MRM)模式,盐酸沙格雷酯质荷比为(m/z 430.1→m/z 135.1)和内标阿立哌唑质荷比为(m/z 448.1→m/z285.1)。结果表明:盐酸沙格雷酯在10.0~1 280.0 ng/m L范围内呈线性,定量限为10.0 ng/m L,基质效应影响小,日内、日间变异系数均小于13.3%,相对回收率在92.0%~118.2%之间。该方法准确、快速、灵敏,可用于微量血浆的盐酸沙格雷酯血药浓度测定、人体内药代动力学及生物等效性研究。

To establish a rapid and accurate method using UPLC-MS/MS for quantification of sarpogrelate hydrochloride in human plasma. One-step protein precipitation with acetonitrile was used to extract the analytes from the plasma. Sarpogrelate and aripiprazole( internal standard, I.S.) were separated on a reversed-phase C18 column within 3.5min, using acetonitrile: ammonium formate( 0.01 mol/L)( 30∶70, ν/ν) as mobile phase at a flow rate of 0.3 m L/min. Mass spectrometry:The analytes were detected by the electronic-spryionization( ESI), Selected multiple reaction monitoring( MRM) using the precursor to product ion combinations of m/z 430.1→135.1 and m/z448.1 →285.1 was performed to detected sarpogrelate hydrochloride and the I.S., respectively.The method was linear over the concentration range of 10.0-1 280.0 ng/m L. The lower limit of quantification was 10 ng/m L. Intra-day inaccuracy was within 13.3% and inter-day was less than12.2%. Matrix effects were minor, the recovery of the method was between 92.0% and 118.2%.The validated method was developed for quantification of sarpogrelate hydrochloride in small volumes of plasma. The method was successfully applied to a pharmacokinetics and bioavailability of sarpogrelate hydrochloride.