Simultaneous Detection Of Six Antituberculosis Drugs In Human Plasma By UPLC-MS/MS

Tuberculosis is an ancient disease. In recent years, the incidence of tuberculosis is rising in the world, and has brought great threat to human life. At present, there are many kinds of first-line antituberculosis drugs, which should be combined to effectively kill mycobacterium tuberculosis in clinical practice. However, the combination of various drugs can bring serious adverse reactions and bacterial drug resistance, which makes the clinician is difficult to determine the drug dosage. How to apply the appropriate dose to achieve the best therapeutic effect and reduce the adverse reactions and drug resistance is still a challenge for the pharmacists and clinicians.At present HPLC-ELSD method and HPLC method are usually used to determine the plasma concentration of antituberculosis drugs which have some help for clinicians to adjust doses of antimicrobial agents. But these methods with higher cost, longer analysis time, lower sensitivity can only detect four kinds of antituberculosis drugs in the most, which limit their values in clinical application.In this research, the UPLC-MS/MS method with lower cost, shorter analysis time and higher sensitivity is adopted to determine the plasma concentration of antituberculosis drugs. Plasma concentration of six kinds of antituberculosis drugs which are ethambutol, isoniazid, pyrazinamide, rifampicin, rifapentine, rifabutin can be detected in the same time. Our study is expected to provide more reliable basis for clinicians in adjusting the dose of antituberculosis drugs.Objective: To evaluate the UPLC-MS/MS method for determining the plasma concentration of antituberculosis drugs and provide more reliable basis for clinicians in adjusting the dose of antituberculosis drugs.Method: The analytes were seperated from the biological matrix by protein precipitation method. Waters ACQUITY UPLC HSS T3(1.8 μm,2.1×100 mm) chromatographic separation column was used for chromatographic fractionation. Acetonitrile-15 m M ammonium and formate-0.05% acid were used for gradient elution with 0.2 m L/min flow rate, 40 degrees centigrade column temperature and 5 μl sample. The ESI source in positive ion mode with multiple reaction monitoring was applied.Results: There were good linear relationship between peak areas and injection quality in the range of 0.05-5,0.1-10,0.2-20, 0.05-10, 0.05-10 and 0.05-5 μg/ml for ethambutol, isoniazid, pyrazinamide, rifampicin, rifapentine and rifabutine respectively with correlation coefficients all greater than 0.991. Exept for pyrazinamide with 15.4% inter-day precision in low concentration, intra-day and inter-day precision were all less than 15% in low, medium and high concentration for other drugs with high accuracy from 88.1 % to 109.1%.Conclusion: The UPLC-MS/MS method with lower cost, shorter analysis time, higher sensitivity and wider linear range can be used to determine the plasma concentration of six antituberculosis drugs simultaneously and can provide more reliable basis for clinicians in adjusting the dose of antituberculosis drugs.