Simultaneous Quantification of Ethambutol and Amikacin in Human Sputum by HPLC-MS/MS / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop an LC-MS method for simultaneously quantifying two anti-tuberculosis drugs (ethambuto land amikacin) in human sputum thus to help guide clinical medication. METHODS: The drugs in sputum were extracted by acetonitrile precipitation, and separated on ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8μm) with mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile (containing 0.1% formic acid) (80∶20) at a flow rate of 0.35 mL•min-1. Multiple reaction monitoring (MRM) was performed in positive ionization mode with electrospray ionization source. RESULTS: The liner calibration curves of ethambutol and amikacin were obtained in the concentration ranges of 48-12 000(r=0.997 8) and 320-80 000(r=0.999 5)ng•mL-1, respectively. The intra- and inter-day precision RSD was less than 9.2% and the absolute recovery was between 96.65% and 106.35%. This method was successfully applied to analyze the drugs in sputum from five clinical samples, the concentration of ethambutol was 148.5 to 919 ng•mL-1, and no amikacin was detected. CONCLUSION: The method is rapid, simple, sensitive and accurate, and can be applied to the therapeutic drug monitoring of ethambutol and amikacin.