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UPLC-MS/MS assay of 21-aminosteroid (lazaroid U74389G) for application in pharmacokinetic study.
J Pharm Biomed Anal. 2016 Apr 15; 122:90-7.JP

Abstract

Lazaroids are potent inhibitors of lipid peroxidation, both in vitro and in vivo. Additionally, a member of the lazaroid family, U-74389G (LAZ) has been shown to have specific radio-protective and anti-proliferative effects. However, there is no quantitative analytical method developed for measuring the therapeutic levels of LAZ for the aforementioned effects. This article highlights the development and validation of a sensitive UPLC-MS/MS method for the quantification of LAZ, and its subsequent application in pharmacokinetic studies in rats with the lower limit of quantification (LLOQ) of 1.95 ng/mL. LAZ and internal standard diadzein (IS) were separated using ACQUITY UPLC(®) BEH C18 column. Gradient elution was used at a flow rate of 0.45 mL/min with mobile phases consisting of 0.1% formic acid in water and 0.1% formic in acetonitrile. LAZ (m/z 612→260) and IS (m/z 255→199) were detected by electrospray ionization (ESI) using multiple reaction monitoring (MRM) in a positive mode on QTRAP(®) 5500 System. The UPLC-MS/MS method was validated as per the US FDA Guidelines for Bio-analytical Validation. LAZ was extracted from rat plasma (100 μL) using protein precipitation by acetonitrile with mean recovery and matrix factor in range of 47.7-56.1%, and 85.6-89.4%, respectively. The calibration curve for LAZ was linear in the range of 1.95-250 ng/mL. The inter-day and intra-day accuracy and precision values for LLOQ, low, medium, high and very high concentration QC samples were within ±15%. LAZ was tested under different storage conditions, for short-term bench-top stability (1h and 3h at 25°C), long-term stability (1 month at -80°C), freeze-thaw cycle stability (1 cycle and 3 cycles) and stability of processed samples in auto-sampler (24h at 10°C) with stability values within ±15% range of nominal concentrations. The validated UPLC-MS/MS method was further applied to a pharmacokinetic study in rats after a single intravenous dose of LAZ at 5 mg/kg.

Authors+Show Affiliations

Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77030, USA. Electronic address: psgadgil@uh.edu.Pfizer, Groton, CT 06340, USA.Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77030, USA. Electronic address: Dchow@uh.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

26848737

Citation

Gadgil, P, et al. "UPLC-MS/MS Assay of 21-aminosteroid (lazaroid U74389G) for Application in Pharmacokinetic Study." Journal of Pharmaceutical and Biomedical Analysis, vol. 122, 2016, pp. 90-7.
Gadgil P, Ibrahim F, Chow DS. UPLC-MS/MS assay of 21-aminosteroid (lazaroid U74389G) for application in pharmacokinetic study. J Pharm Biomed Anal. 2016;122:90-7.
Gadgil, P., Ibrahim, F., & Chow, D. S. (2016). UPLC-MS/MS assay of 21-aminosteroid (lazaroid U74389G) for application in pharmacokinetic study. Journal of Pharmaceutical and Biomedical Analysis, 122, 90-7. https://doi.org/10.1016/j.jpba.2016.01.033
Gadgil P, Ibrahim F, Chow DS. UPLC-MS/MS Assay of 21-aminosteroid (lazaroid U74389G) for Application in Pharmacokinetic Study. J Pharm Biomed Anal. 2016 Apr 15;122:90-7. PubMed PMID: 26848737.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - UPLC-MS/MS assay of 21-aminosteroid (lazaroid U74389G) for application in pharmacokinetic study. AU - Gadgil,P, AU - Ibrahim,F, AU - Chow,D S-L, Y1 - 2016/01/16/ PY - 2015/10/16/received PY - 2016/01/09/revised PY - 2016/01/12/accepted PY - 2016/2/6/entrez PY - 2016/2/6/pubmed PY - 2016/12/15/medline KW - 21-Aminosteroid KW - LC–MS KW - Stability KW - U-74389G SP - 90 EP - 7 JF - Journal of pharmaceutical and biomedical analysis JO - J Pharm Biomed Anal VL - 122 N2 - Lazaroids are potent inhibitors of lipid peroxidation, both in vitro and in vivo. Additionally, a member of the lazaroid family, U-74389G (LAZ) has been shown to have specific radio-protective and anti-proliferative effects. However, there is no quantitative analytical method developed for measuring the therapeutic levels of LAZ for the aforementioned effects. This article highlights the development and validation of a sensitive UPLC-MS/MS method for the quantification of LAZ, and its subsequent application in pharmacokinetic studies in rats with the lower limit of quantification (LLOQ) of 1.95 ng/mL. LAZ and internal standard diadzein (IS) were separated using ACQUITY UPLC(®) BEH C18 column. Gradient elution was used at a flow rate of 0.45 mL/min with mobile phases consisting of 0.1% formic acid in water and 0.1% formic in acetonitrile. LAZ (m/z 612→260) and IS (m/z 255→199) were detected by electrospray ionization (ESI) using multiple reaction monitoring (MRM) in a positive mode on QTRAP(®) 5500 System. The UPLC-MS/MS method was validated as per the US FDA Guidelines for Bio-analytical Validation. LAZ was extracted from rat plasma (100 μL) using protein precipitation by acetonitrile with mean recovery and matrix factor in range of 47.7-56.1%, and 85.6-89.4%, respectively. The calibration curve for LAZ was linear in the range of 1.95-250 ng/mL. The inter-day and intra-day accuracy and precision values for LLOQ, low, medium, high and very high concentration QC samples were within ±15%. LAZ was tested under different storage conditions, for short-term bench-top stability (1h and 3h at 25°C), long-term stability (1 month at -80°C), freeze-thaw cycle stability (1 cycle and 3 cycles) and stability of processed samples in auto-sampler (24h at 10°C) with stability values within ±15% range of nominal concentrations. The validated UPLC-MS/MS method was further applied to a pharmacokinetic study in rats after a single intravenous dose of LAZ at 5 mg/kg. SN - 1873-264X UR - https://www.unboundmedicine.com/medline/citation/26848737/UPLC_MS/MS_assay_of_21_aminosteroid__lazaroid_U74389G__for_application_in_pharmacokinetic_study_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0731-7085(16)30032-2 DB - PRIME DP - Unbound Medicine ER -