高效液相色谱-串联质谱法测定人血浆中消旋卡多曲活性代谢物含量

Quantitative Determination of Active Racecadotril Metabolites in Human Plasma with High Performance Liquid Chromatography Tandem Mass Spectrometry

建立高效液相色谱-串联质谱法（HPLC-MS/MS）测定人血浆中消旋卡多曲活性代谢物Thiorphan（TP）的含量及S-Methyl-Thiorphan（MTP）的相对含量。采用Kromasil C18色谱柱（25mm×4.6mm,5μm）,流动相为V（水）∶V（乙腈）=72∶28,甲酸调pH至2.8。加抗氧化剂的血浆样品经乙腈直接沉淀后进样、MRM模式外标法定量,TP和MTP检测离子对分别为m/z252/218和m/z266/218。TP在6.25～3200μg/L范围内线性关系良好（r=0.9998）;方法回收率为97.1%～109.0%,批内批间RSD分别小于3.5%及6.0%。

A high performance liquid chromatography-tandem mass spectrometric （HPLC-MS/MS） method was established for the determination of the racecadotril metabolites, thiorphan （TP） and S-methyl-thiorphan （MTP） in human plasma. Kromasil C18 column （25 mm ×4.6 mm, 5 μm） was used. The mobile phase was water-acetonitrile （72：28, pH 2. 8 with formic acid）. The plasma added antioxidant was precipitated with acetonitrile and the content of drug was determined by HPLC-MS/MS using electrospray ionization. TP and MTP were detected on multiple reaction monitoring mode by the transitions from the precursor to the product ion （m/z 252/218 and m/z 266/218）. For TP, calibration curve was linear in the range from 6.25 to 3200 μg/L, the limit of quantitation in plasma was 6.25 μg/L, the method recovery was 97.1% - 109.0% , the RSD of inter-and intra-day precision were less than 3.5% and 6.0% , respectively.