Objective: To develop a high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the determination of vildagliptin in human anticoagulant plasma with ethylenediamine tetra acetic acid and apply it to the study of pharmacokinetics. Methods: 13C-15N-vildagliptin was used as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all components were separated by a Hypurity C18 column (150 mm×2.1 mm,5 μm), using a gradient elution procedure consisting of methanol and 5 mmol·L-1 ammonium formate at a flow rate of 0.5 mL·min-1,and the column temperature was 40 ℃. Injection volume was just 2 μL. Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 304.3→154.2 for vildagliptin and m/z 310.3→160.3 for internal standard. Specificity,standard curve, lower limit of quantification,precision,recovery,matrix effect and stability were examined. Then this method was used to determine the plasma concentration of veragliptin in healthy subjects. Results: The calibration curve of vildagliptin in human plasma was linear over the concentration range of 1.11 to 534.0 ng·mL-1. The lower limit of quantitation was 1.11 ng·mL-1. The intra-and inter-day precisions at four quality control levels were within 0.9%-8.5%,and the accuracy was within 99.8%-109.3%. The data of short-term stability at room temperature displayed that the accuracy percentage of LQC samples was 92.0% for 0.5 h exposure, 87.6% for 1 h exposure, 71.2% for 2 h exposure. These of LQC samples chilled on ice was 102.0% for 0.5 h exposure, 94.5% for 1 h exposure, 86.6% for 2 h exposure. These results showed a phenomenon that there was a possible degradation of vildagliptin in plasma. The results of extraction recovery and matrix effect and other stability met the requirements of biological sample analysis. The pharmacokinetic study results of 8 healthy subjects showed that t1/2 was (1.49±0.37) h, tmax was (2.06±1.11) h, Cmax was (290.94±100.36) ng·mL-1, AUC0-24 h was (1 343.46±186.89) ng·h·mL-1, AUC0-∞ was (1 351.31±188.79) ng·h·mL-1. Conclusion: This method is easy to operate, has high specificity, and sensitivity. It has been successfully applied to the pharmacokinetic study of 8 healthy subjects after oral administration of 50 mg vigagliptin tablets on an empty stomach. Therefore, it can be used as a reliable detection method for human pharmacokinetic research and therapeutic drug monitoring.
WANG Yi-cheng, HE Kang, PENG Jing-bo, RAO Tai, CHEN Yao, GUO Ying, TAN Zhi-rong
. HPLC-MS/MS method for the quantification of vildagliptin in human plasma and its application*[J]. Chinese Journal of Pharmaceutical Analysis, 2024
, 44(1)
: 68
-75
.
DOI: 10.16155/j.0254-1793.2024.01.07
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