High Performance Liquid Chromatographic Determination of Anti-Tuberculosis Drugs in Human Body Fluids

Abstract

A rapid, simple and sensitive high performance liquid chromatographic (HPLC) assay for the quantification of pyrazinamidum (PZA), rimifon (INH) and rifapentine (RFT) in human serum, urine and cerebrospinal fluid (CSF) has been developed. After acidification and removal of protein, liquid samples were injected into the HPLC system directly. Separation was achieved using a μBondapak phenyl reversed phase column for RFT and ion-pair chromatography on Nova-pak C₁₈ for INH and PZA. The retention times of PZA and INH on the -C₁₈ column were 2.66 and 5.33 min, respectively. Retention time of RFT was 9.10 min. on the -phenyl column. The PZA, INH and RFT standard plots were highly linear (r>0.99) over the concentration range of 0.1 to 10 μg/mL, 0.1 to 10 μg/mL and 0.5 to 10 μg/mL. respectively. PZA mean recovery was 96.5%±4.2, INH 97.4%±3.7 and RFT 92.1%±6.2. The method was applied for the control of the drug doses on treating tuberculous meningitis and for investigating whether RFT passes through human hematoencephalic barrier.