液相色谱串联质谱法测定人血清中匹多莫德的浓度

Determination of Pidotimod in Human Serum by HPLC-MS/MS

目的:建立液相色谱串联质谱法测定人血清中匹多莫德的浓度。方法:血清样品用甲醇沉淀蛋白,内标为格列吡嗪,色谱柱为LichrospherC18(4.6mm×150mm,5μm),柱温为30℃,流动相为甲醇-2.5mmol/L NH4Ac(90∶10,V/V),流速为0.5mL/min。质谱采用ESI离子源负离子检测,定量分析的离子对为:m/z 242.9/153.0(匹多莫德),m/z 444.0/169.9(内标格列吡嗪)。结果:匹多莫德在0.1～10mg/L范围内线性关系良好(r=0.9995),批内批间RSD均小于15%,提取回收率为85.52%～98.30%。结论:该方法快速,灵敏,准确,可用于匹多莫德的临床药动学研究。

To develop and validate an HPLC-MS/MS for the determination of pidoti- mod in human serum. METHODS. Glipizide was added as internal standard. The sample prepara- tion included a simple deproteinization step withmethanol. Chromatographic separation was a- chieved on a Lichrospher C18 column（4.6 mm）〈 150 mm, 5 μm）at 30 ℃ with the mobile phase consisted of methanol-2.5 mmol/L NH4Ac （90 ： 10, V/V）. The mobile phase was eluted at aflow rate of 0.5 mL/min. Tandem mass spec- trometer equipped with electrospray ionization source was applied and operated in the negative ion mode. Multiple reaction monitoring using the precursor to product ion combinations of m/ z 242.9/153.0 and m/z 444.0/169.9 was per- formed to quantify pidotimod and glipizide, re- spectively. RESULTS. The calibration curves were linear over the range of 0.1--10 mg/L（r= 0. 9995）. RSD of intra-batch and inter-batchwere less than 15%, extraction recovery of three level concentrations were 85.52%- 98.30%. CONCLUSION. The method is shown to be con- venient, accurate, sensitive, and suitable for clinical pharmacokinetic study of pidotimod.