Simultaneously Quantifying IsoniazidPyrazinamide and Levofloxacin in Human Sputum by HPLC-MS/MS / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop an UPLC-MS/MS method for simultaneous determination of 3 anti-tuberculosis drugs [isoniazid (INH) pyrazinamide (PZA) and levofloxacin(LFX)] in sputum by HPLC-MS/MS. METHODS The drugs(INH,PZAand LFX)were extracted by methanol/acetonitrile (V-V=1∶1). All of them separated on ACQUITY UPLC HSS T3 1.8 μm column (2.1 mm×100 mm,Waters Corporation) with the mobile phase of aqueous solution (containing 0.1% formic acid)-acetonitrile (containing 0.1% formic acid) at a flow rate of 0.35 mL•min-1. Multiple reaction monitoring (MRM) mode was performed combined with electrospray ionization source operating in the positive ionization mode. Furthermore, it was blindly collected 7 sputum samples, and analyzed the drug concentration by the built method in this work. RESULTS: The liner calibration curves of INHPZALFX were obtained in the concentration range of 48-6 000 ng•mL-1(r=0.998 8)480-60 000 ng•mL-1(r=0.999 3)120-15 000 ng•mL-1(r=0.999 5), respectively. The CV of intra- and inter-day precision was less than 15% and the absolute recovery was 97.21%-107.80%. In the 7 clinical samples, the concentration of INH is 44.74, 120.1, 301.5, 481, 595.5, 1 220 and 1 570 ng•mL-1, 104.2, 6 273.34 and 3 185 ng•mL-1 for PZA, and 199.86 for LFX, respectively. CONCLUSION: In this work, it is built a LC-MS method to simultaneously quantify INH, PZA and LFX, which is simple, sensitive and accurate. This method is successfully applied to analyze the clinical samples.