超高效液相色谱-质谱法测定大鼠血浆中的姜黄素

Determination of Curcumin in Rat Plasma by UPLC-MS/MS

建立一个测定大鼠血浆中姜黄素含量的液相色谱串联三重四级杆质谱法(UPLC-MS/MS).血浆样品用甲醇沉淀蛋白,采用Agilent Extend-C18色谱柱(RRHD 1.8μm,2.1 mm×50 mm),以乙腈(A)-0.1%甲酸水(B)为流动相梯度洗脱,流速为0.2 m L/min,柱温30℃,质谱条件为电喷雾离子源(ESI),正离子模式,内标为沙丁胺醇,采用多反应监测(MRM)模式测定,用于定量分析的离子对:姜黄素和内标分别为369.1→177.1和240.1→148.姜黄素在2.5~400 ng/m L范围内线性关系良好,最低检出限为0.084 ng/m L,提取回收率在84.66%~95.71%,日内及日间精密度RSD均小于7%(n=3),稳定性较好.该方法准确、高效、专属性强、灵敏度高、测定结果准确可靠,可用于实际大鼠血浆中姜黄素的含量测定,为监测姜黄素口服给药后体内的血药浓度和药代动力学特征提供参考,也为其新制剂的设计和临床应用提供数据基础.

A method for the determination of curcumin in rat plasma was established by liquid chromatography-tandem quadrupole mass spectrometry(UPLC-MS/MS). Plasma samples were precipitated with methanol andeluted with an Agilent Extend-C18 column(RRHD 1.8 μm,2.1 mm×50 mm)and eluting with acetonitrile(A)-0.1%formic acid(B)as the mobile phase gradient at a flow rate of 0.2 m L/min,and the column temperature was 30 ℃.The mass spectrometric conditions were electrospray ionization(ESI),positive ion mode,and the internal standard(IS)was salbutamol. Detection was carried out by multiple reaction monitoring(MRM)of the transitions(precursorion to product ion)at m/z 369.1 to 177.1 for curcumin,m/z 240.1 to 148 for IS. The linear range was 2.5 ng/m L to400 ng/m L,the limit of detection of curcumin was 0.084 ng/m L,the recoveries were 84.66% to 95.71%,and theRSD of intra and inter-day precisions were less than 7%(n=3). The stability was better. The method was accurate,efficient,specific and high sensitive. The determination result was accurate and reliable. It could be used for thedetermination of curcumin in rat plasma and to provide reference for the monitoring concentration and pharmacokineticcharacteristics of curcumin in rat plasma after oral administration,and to provide a theoretical basis for the designand clinical application of the new formulations.