Simultaneously Quantifying Isoniazid、Pyrazinamide and Levofloxacin in Human Sputum by HPLC-MS/MS
SUN Tao1, ZHENG Ze1,2, XI Xiu-hong1, LI Tao1, JIA Xiao-fang1, JIAO Ping2, YIN Lin1, ZHANG Li-jun1*
1. Shanghai Public Health Clinical Center, Shanghai 201508, China; 2. School of Chemistry and Chemical Engineering, Hubei Polytechnic University, Huangshi 435003, China
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,PZA、and 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 INH、PZA、LFX 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.
SOTGIU G, SULIS G, MATTEELLI A. Tuberculosis world health organization perspective[J]. Microbiol Spectr, 2017, 5(1):doi:10.1128/microbiolspec.TNMI7-0036-2016.
[2]
PASIPANODYA J G, MCILLERON H, BURGER A, et al. Serum drug concentrations predictive of pulmonary tuberculosis outcomes[J]. J Infect Dis, 2013, 208(9):1464-1473.
[3]
FEYISA S G, ABDURAHMAN A A, JIMMA W, et al. Resistance of Mycobacterium tuberculosis strains to rifampicin:a systematic review and Meta-analysis[J]. Heliyon, 2019, 5(1):e1081. doi:10.1016/j.heliyon.2018.e01081. eCollection 2019 Jan.
[4]
WU B, ZHANG L, LIU Z, et al. Drug-resistant tuberculosis in Zhejiang Province, China:an updated analysis of time trends, 1999-2013[J]. Glob Health Action, 2017, 10(1):1293925. doi:10.1080/16549716.2017.1293925.
[5]
DARTOIS V, BARRY C E. Clinical pharmacology and lesion penetrating properties of second- and third-line antituberculous agents used in the management of multidrug-resistant (MDR) and extensively-drug resistant (XDR) tuberculosis[J]. Curr Clin Pharmacol, 2010, 5(2):96-114.
[6]
DHEDA K, GUMBO T, MAARTENS G, et al. The epidemiology, pathogenesis, transmission, diagnosis, and management of multidrug-resistant, extensively drug-resistant, and incurable tuberculosis[J]. Lancet Respir Med, 2017:doi:10.1016/S2213-2600(17)30079-6. [Epub ahead of print].
[7]
ABBARA A, CHITTY S, ROE J K, et al. Drug-induced liver injury from antituberculous treatment:a retrospective study from a large TB centre in the UK[J]. BMC Infect Dis, 2017, 17(1):231. doi:10.1186/s12879-017-2330-z.
[8]
ZHANG Y, WU S, XIA Y, et al. Adverse events associated with treatment of multidrug-resistant tuberculosis in China: an ambispective cohort study[J]. Med Sci Monit, 2017, 23:2348-2356.
[9]
ALSULTAN A, PELOQUIN C A. Therapeutic drug monitoring in the treatment of tuberculosis:an update[J]. Drugs, 2014, 74(8):839-854.
[10]
VERBEECK R K, GUNTHER G, KIBUULE D, et al. Optimizing treatment outcome of first-line anti-tuberculosis drugs:the role of therapeutic drug monitoring[J]. Eur J Clin Pharmacol, 2016, 72(8):905-916.
[11]
PELOQUIN C. The role of therapeutic drug monitoring in mycobacterial infections[J]. Microbiol Spectr, 2017, 5(1). doi:10.1128/microbiolspec.TNMI7-0029-2016.
[12]
BALDWIN D R, HONEYBOURNE D, WISE R. Pulmonary disposition of antimicrobial agents:methodological considerations[J]. Antimicrob Agents Chemother, 1992, 36(6):1171-1175.
[13]
BRUSSE-KEIZER M, TEN B L, MOVIG K, et al. Relation between amoxicillin concentration in sputum of COPD patients and length of hospitalization[J]. COPD, 2011, 8(2):66-70.
[14]
BRUSSE-KEIZER M, VANDERVALK P, VAN DER ZANDEN R W, et al. Amoxicillin concentrations in relation to beta-lactamase activity in sputum during exacerbations of chronic obstructive pulmonary disease[J]. Int J Chron Obstruct Pulmon Dis, 2015, 10:455-461.
[15]
SISTIK P, TURJAP M, IORDACHE A M, et al. Quantification of selected antidepressants and antipsychotics in clinical samples using chromatographic methods combined with mass spectrometry: a review (2006-2015)[J]. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 2016, 160(1):39-53.
[16]
MOTA L, AL-EFRAIJ K, CAMPBELL J R, et al. Therapeutic drug monitoring in anti-tuberculosis treatment:a systematic review and Meta-analysis[J]. Int J Tuberc Lung Dis, 2016, 20(6):819-826.
2020, Vol. 55 
