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An UHPLC-UV Method for the Determination of Vancomycin in Human Serum

Author(s):

Fang Fang*, Ning Li, Chunli Xu, Rong Tan, Jihong Yang and Zhichang Zheng   Pages 1 - 6 ( 6 )

Abstract:


Objective: To develop a rapid ultra-performance liquid chromatographic (UHPLC)-UV method for vancomycin determination in human serum for therapeutic drug monitoring (TDM).

Methods: Human serum samples were precipitated with 10% perchloric acid, and the supernatant after centrifugation was analyzed on an ACQUITY UHPLC BEH C18 column (2.1 × 50mm, 1.7 μm) via gradient elution with a flow rate at 0.3 mL/min. The mobile phase consisted of acetonitrile and 0.005M KH2PO4 buffer (containing 0.1% triethylamine, pH 3.4). The detection wavelength was set at 210 nm, and the column temperature was set at 40. The total runtime was 6.0 min per analysis.

Results: After comprehensive validation, the method was applied to determine the concentration of vancomycin in human serum. The chromatographic peaks of vancomycin and internal standard were not interfered by endogenous matrixes. The retention time (RT) of vancomycin was 1.91 min, while the internal standard was 1.58 min. The good linearity range of vancomycin concentration was 2.5-120 μg/mL (R2>0.999). The lower limit of quantitation (LLOQ) was 2.5 μg/mL. The precision at three quality control (QC) levels (including LLOQ) was restricted within 85-115%. The extraction recovery rate of QC samples (4.0, 20.0, 60.0 μg/mL) were 101.16%、97.70%、94.90%, respectively. Inter- and intra-day precision was less than 8% (RSD). Stability tests under different storage conditions were satisfactory. In patients, the concentration of vancomycin ranged from 7.30 to 89.12 μg/mL determined by the fully validated method.

Conclusion: The simple, rapid sample pretreatment procedures and short analysis time made this UHPLC-UV method suitable for therapeutic drug monitoring (TDM) of vancomycin.

Keywords:

vancomycin, ultra-performance liquid chromatographic (UHPLC) therapeutic drug monitoring

Affiliation:

Department of Pharmaceutics, Affiliated Hospital of Guizhou Medical University, Guiyang, 550001, National Experimental Teaching Demonstration Center of Pharmacy, China Pharmaceutical University, Nanjing, 211198, Department of Pharmaceutics, Affiliated Hospital of Guizhou Medical University, Guiyang, 550001, Department of Pharmaceutics, Affiliated Hospital of Guizhou Medical University, Guiyang, 550001, Department of Pharmaceutics, Affiliated Hospital of Guizhou Medical University, Guiyang, 550001, Department of Pharmaceutics, Affiliated Hospital of Guizhou Medical University, Guiyang, 550001



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