title: Quantification of levornidazole and its metabolites in human plasma and urine by ultra-performance liquid chromatography-mass spectrometry.
abstract: We developed and validated an ultra-performance liquid chromatographic (UPLC) method coupled with atmospheric pressure chemical ionization (APCI) mass spectrometry for simultaneous determination of levornidazole and its first-pass metabolites, l-chloro-3-(2-hydroxymethyl-5-nitro-l-imidazolyl)-2-propanol (Ml), 2-methyl-5-nitroimidazole (M2) and 3-(2-methyl-5-nitro-1-imidazolyl)-1,2-propanediol (M4), in human plasma and urine. The biological samples were pretreated by protein precipitation and liquid-liquid extraction and analyzed using an ACQUITY UPLC CSH C18 column (2.1×50 mm, 1.7 μm) and a QTRAP mass spectrometer in multiple reaction monitoring mode via APCI. Acetonitrile and 0.1% formic acid in water was used as the mobile phase in gradient elution at a flow rate of 0.6 mL/min. The lower limit of quantification of this method was 0.0100, 0.00500, 0.0200 and 0.00250 μg/mL for levornidazole, M1, M2 and M4, respectively. The linear calibration curves were obtained for levornidazole, M1, M2, and M4 over the range of 0.0100-5.00, 0.00500-2.50, 0.0200-10.0 and 0.00250-1.25 μg/mL, respectively. The intra- and inter-batch precision was less than 12.2% in plasma and less than 10.8% in urine. The intra- and inter-batch accuracy was 87.8-105.7% in plasma and 92.8-109.2% in urine. The mean recovery of levornidazole, M1, M2 and M4 was 91.1-105.1%, 95.8-103.8%, 87.8-96.8%, 96.8-100.6% from plasma and 96.0-100.9%, 96.9-107.9%, 95.1-102.7%, 103.7-105.9% from urine respectively. This method was validated under various conditions, including room temperature, freeze-thaw cycles, long-term storage at -40 ± 5°C, after pretreatment in the autosampler (at 10°C), and 10- and 100-fold dilution. This newly established analytical method was successfully applied in a pharmacokinetic study following single intravenous infusion of levornidazole in 24 healthy Chinese subjects.
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Yuran
authors: Y
authors: Cao
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Miao
authors: M
authors: Zhao
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Xiaojie
authors: X
authors: Wu
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Beining
authors: B
authors: Guo
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Yuancheng
authors: Y
authors: Chen
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Jicheng
authors: J
authors: Yu
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Guoying
authors: G
authors: Cao
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China. Electronic address: zhangj_fudan@aliyun.com.
authors: Jing
authors: J
authors: Zhang
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Yaoguo
authors: Y
authors: Shi
authors: Institute of Antibiotics, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China; Key Laboratory of Clinical Pharmacology of Antibiotics, National Health and Family Planning Commission, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors: Yingyuan
authors: Y
authors: Zhang
mesh_terms: D000981:Antiprotozoal Agents / Q000097:blood* / Q000737:chemistry / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D007536:Isomerism; D057230:Limit of Detection; D008297:Male; D009950:Ornidazole / Q000031:analogs & derivatives / Q000097:blood* / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000981:Antiprotozoal Agents; D009950:Ornidazole
keywords: LC–MS/MS; Levornidazole; Metabolite; Pharmacokinetics