UPLC-MS/MS法测定大鼠血浆中的新型促尿酸排泄剂TJ-D-1

Establishment and validation of an UPLC-MS/MS method to quantify a new uricosuric compound TJD-1 in rat plasma

目的建立一种超高效液相色谱-串联质谱(UPLC-MS/MS)法测定大鼠血浆中TJ-D-1的质量浓度。方法取血浆样品50μL以乙腈沉淀蛋白处理,取上清液用水稀释(体积比1∶1),TJ-D-1和内标(苯溴马隆)经ACQUITY UHPLC BEH C18(50 mm×2. 1 mm I. D,1. 7μm)色谱柱分离,以乙腈-水(体积比60∶40)为流动相等度洗脱,色谱运行时间为3. 0 min。质谱采用电喷雾电离源(ESI)负离子模式,扫描方式为多反应监测(MRM)。用于定量检测的离子对分别为m/z 440. 7→250. 9(TJ-D-1)和m/z 423. 2→250. 9(苯溴马隆)。结果大鼠血浆中TJ-D-1方法的线性范围为0. 1～50 mg·L-1,日内、日间精密度RSD均小于12. 7%,准确度(RE)为-2. 8%～1. 8%,提取回收率为94. 8%～110. 7%,内标归一化的基质因子为100%～107%,且相对标准偏差(RSD)均小于8. 0%。结论该法适用于测定大鼠血浆中TJ-D-1的质量浓度,为TJ-D-1的临床前药物动力学研究提供科学参考。

TJ-D-1 is a newuricosuric agent and can significantly reduce serum uric acid in hyperuricemic rats. Objective To establish an ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for the simple,accurate and rapid determination of TJ-D-1 in rat plasma. Methods The plasma sample was extracted by protein precipitation with acetonitrile and the supernatant was diluted with water(1 ∶ 1,V ∶ V). TJ-D-1 and the internal standard benzbromarone were separated on a ACQUITY UHPLC BEH C18(2. 1 mm × 50 mm I. D,1. 7 μm) column with an isocratic mobile phase consisting of acetonitrile-water( 60 ∶ 40,V ∶ V). The total run time was 3. 0 min. Negative electrospray ionization was performed using multiple reaction monitoring( M RM) with transitions of m/z 440. 7→250. 9 for TJ-D-1 and m/z 423. 2→250. 9 for IS. Results The standard curve was linear in the range of 1. 00 × 102-5. 00 ×104 ng·mL-1 with intra-and inter-day precision within 12. 7% and accuracy in range of-2. 8% to 1. 8%.The average recoveries were between 94. 8% and 110. 7%. The matrix effects were between 100% and107% and the relative standard deviations( RSD) were less than 8. 0%. Conclusion This UPLC-M S/M S method is simple,accurate and rapid for quantifying TJ-D-1 in rat plasma,which will lay the foundation for the preclinical study.