Simultaneous determination of isoniazid, rifampicin, levofloxacin in mouse tissues and plasma by high performance liquid chromatography-tandem mass spectrometry.
A rapid and selective high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneous determination of isoniazid (INH), rifampicin (RFP) and levofloxacin (LVX) in mouse tissues and plasma has been developed and validated, using gatifloxacin as the internal standard (I.S.). The compounds and I.S. were extracted from tissue homogenate and plasma by a protein precipitation procedure with methanol. The HPLC separation of the analytes was performed on a Welch materials C4 column (250mmx4.6mm, 5.0microm, USA) at 25 degrees C, using a gradient elution program with the initial mobile phase constituting of 0.05% formic acid and methanol (93:7, v/v) at a flow-rate of 1.0ml/min. For all the three analytes, the recoveries varied between 83.3% and 98.8% in tissues and between 75.5% and 90.8% in plasma, the accuracies ranged from 91.7% to 112.0% in tissues and from 94.6% to 108.8% in plasma, and the intra- and inter-day precisions were less than 13.3% in tissues and less than 8.2% in plsama. Calibration ranges for INH were 0.11-5.42microg/g in tissues and 0.18-9.04microg/ml in plasma, for RFP were 0.12-1200microg/g in tissues and 4.0-200microg/ml in plasma, and for LVX were 0.13-26.2microg/g in tissues and 0.09-4.53microg/ml in plasma. The lower limits of quantification (LLOQs) for INH, RFP and LVX in mouse tissues were 0.11, 0.12 and 0.13microg/g and for those in mouse plasma were 18.1, 20.0 and 21.8ng/ml, respectively. The limits of detection (LODs) for INH, RFP and LVX in mouse tissues were 0.04, 0.05 and 0.05microg/g and for those in mouse plasma were 5.5, 6.0 and 6.6ng/ml, respectively. The established method was successfully applied to simultaneous determination of isoniazid, rifampicin and levofloxacin in mouse plasma and different mouse tissues.
Fang
Ping-Fei
PF
Clinical Pharmacy and Pharmacology Research Institute, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Cai
Hua-Lin
HL
Li
Huan-De
HD
Zhu
Rong-Hua
RH
Tan
Qin-You
QY
Gao
Wei
W
Xu
Ping
P
Liu
Yi-Ping
YP
Zhang
Wen-Yuan
WY
Chen
Yong-Chang
YC
Zhang
Feng
F
D000818:Animals; D000904:Antibiotics, Antitubercular / Q000032:analysis / Q000097:blood; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D024841:Fluoroquinolones / Q000737:chemistry; D000077734:Gatifloxacin; D007421:Intestine, Small / Q000737:chemistry; D007538:Isoniazid / Q000032:analysis / Q000097:blood; D007668:Kidney / Q000737:chemistry; D016018:Least-Squares Analysis; D064704:Levofloxacin; D008099:Liver / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D008297:Male; D000432:Methanol / Q000737:chemistry; D051379:Mice; D015242:Ofloxacin / Q000032:analysis / Q000097:blood; D015203:Reproducibility of Results; D012293:Rifampin / Q000032:analysis / Q000097:blood; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014018:Tissue Distribution
D000904:Antibiotics, Antitubercular; D024841:Fluoroquinolones; D064704:Levofloxacin; D015242:Ofloxacin; D000077734:Gatifloxacin; D007538:Isoniazid; D012293:Rifampin; D000432:Methanol