Validation of Pyrazinamide in Human Plasma Using HplcUv for Therapeutic Drug Monitoring

Currently, Indonesia is in the 5 th rank of the highest tuberculosis prevalence over the world. The treatment of tuberculosis is going complicated due to the side effect experienced by the patients. The four combination of antituberculosis agent used in minimally 6 months of treatment could stimulate the hepatotoxicity as the one of the dominant side effect in tuberculosis treatment. Thus, it is important to do the Therapeutic Drug Monitoring (TDM) to optimize the tuberculosis treatment. This study is aimed to validate the TDM of pyrazinamide in human plasma using High Performance Liquid Chromatography-UV. We recruited 6 TB patients in the validation of pyrazinamide study. The C18 column shim- pack VP-ODS (250 mm x 4.6 mm, id 5μm) and aquabidest-acetonitrile as mobile phase were applied in this study. We used Shimadzu HPLC system with a model AT LC20 LC 10AT pump, detector SPD 20A and LC solution software. We performed the analysis for linearity, system appropriateness, accuracy and recovery to develop the validation method. This study has been approved by National Ethics Committee of Health Research. Our study shows that the linearity is good with value of r2 > 0.99 and the equation y = 16740.876x - 2953.615. The CV TR and CV peak area for system suitability are 1.46% and 0.29%, respectively. The LoD and LoQ value are 2.532 and 7.672 μg/mL, respectively. The accuracy on the concentration of 1.00, 8.00, 60.00 ug/ml are 108.80 %, 92.57 % and 100.98 %, respectively for intraday accuracy and 103.18 %, 92.44%, and 94.94%, respectively for interday accuracy. Furthermore, the precision on the concentration of 1.00, 8.00, 60.00 ug/ml are 1.17%, 3.57%, 3.32%, respectively for intraday precision and 3.66%, 1.37% and 1.59%, respectively for interday precision. In conclusion, the method which we applied in this study was sensitive and reliable for routine TDM of pyrazinamide