Establishment and application of HPLC in determining INH, AcINH, RFP and PZA simultaneously in plasma of patients with tuberculosis
ZHANG Liang
QIAN Zhi-lei
LU Lei
HOU Wen-jie
DING Hong-zi
WANG Jie
Abstract：Objective To establish the high-performance liquid chromatography ( HPLC) in order to determine the concentration of isoniazid (INH), rifampicin (RFP), pyrazinamide (PZA) and acetylisoniazid (AcINH) simultaneously in the plasma of patients with tuberculosis , and to verify its effect .Methods The HPLC method was carried out on a Wa-ters Xselect-HSS T3 (250 mm ×4.6 mm, 5 μm) column with 0.05 mol/L KH2 PO4 ( adjusting PH to 5.6 with triethyl-amine)-acetonitrile as mobile phase .The column temperature was 40 ℃, the flow rate was 0.8-1.0 mL/min, and the in-ject volume was 20μL.Meanwhile, the plasma containing reference substances of INH , RFP, PZA and AcINH were injec-ted.Then we tested the specificity , linear relationship, precision, accuracy, stabilization and extraction recovery .We de-tected the concentrations of INH , AcINH, RFP and PZA in the plasma of 10 tuberculosis patients who took INH , RFP and PZA as treatment program by HPLC method .Results This method had a good separation effect on INH , AcINH, RFP, and PZA without interference of each other , and all calibration curves had good linearity ( r2 >0.99) within the test ran-ges.The intra-and inter-day precision was less than 10%, the extraction recovery was between 80%-120%.Except 1 case of AcINH and 1 case of RFP below the linear range , during the 10 tuberculosis patients , the rest were within the linear range.Conclusions HPLC that can determine the concentrations of INH , PZA, RFP and AcINH simultaneously in plas-ma of tuberculosis patients is successfully established .The method is sensitive , simple and rapid .It is suitable for analyti-cal require of biological samples .
Keywords：tuberculosishigh-performance liquid chromatography (HPLC)isoniazidrifampicinpyrazinamideacetyli-soniazidblood drug concentration
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 23-26 )