Automated high-performance liquid chromatography method for the determination of rosiglitazone in human plasma |
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| Institution: | 1. Leeds Institute of Rheumatic and Musculoskeletal Medicine, St. James Hospital, University of Leeds, UK;2. NIHR Biomedical Research Unit, Chapel Allerton Hospital, University of Leeds, UK;3. Clinical Pathology Department, Faculty of Medicine, Mansoura University, Egypt;1. Low Carbon Energy Institute and School of Chemical Engineering, China University of Mining and Technology, Xuzhou, 221008, People''s Republic of China;2. School of Science, City University of Hong Kong, Hong Kong SAR 999077, People''s Republic of China;3. Department of Physics, Sungkyunkwan University, Suwon 16419, Republic of Korea;4. School of Science, Xi''an Polytechnic University, Xi''an 710048, People''s Republic of China;5. School of Environment and Safety Engineering, North University of China, Taiyuan 030051, People''s Republic of China;1. Department of Pharmacology, School of Pharmaceutical Education and Research, Jamia Hamdard (Hamdard University), New Delhi, 110062, India;2. Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow, 226031, India;3. Department of Pharmacology, Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Pune, 411038, India;1. State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, China;2. Biomaterials & Tissue Engineering Division, Department of Endodontics, Prosthodontics and Operative Dentistry, University of Maryland Dental School, Baltimore, MD 21201, USA;3. Center for Stem Cell Biology and Regenerative Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA;4. Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA;5. Mechanical Engineering Department, University of Maryland, Baltimore County, MD 21250, USA |
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| Abstract: | A robust, fully automated assay procedure for the determination of rosiglitazone (I, BRL-49653) in human plasma has been developed. Plasma concentrations of I were determined using automated sequential trace enrichment of dialysates (ASTED) coupled to reversed-phase high-performance liquid chromatography. Sequential automated dialysis of human plasma samples was followed by concentration of the dialysate by trace enrichment on a C18 cartridge. Drug and internal standard, SB-204882 (II) were eluted from the trace enrichment cartridge by mobile phase (0.01 M ammonium acetate, pH 8–acetonitrile, 65:35, v/v) onto the HPLC column (a Novapak C18, 4 μm, 100×5 mm radial compression cartridge) protected by a Guard-Pak C18 cartridge. The compounds were detected by fluorescence detection, using an excitation wavelength of 247 nm, and emission wavelength of 367 nm. The lower limit of quantitation of the method was 3 ng/ml (200 μl aliquot) with linearity demonstrated up to 100 ng/ml. Within- and between-run precision and accuracy of determination were better than 10% across the calibration range. There was no evidence of instability of I in human plasma following three complete freeze–thaw cycles and samples can be safely stored for at least 7 months at −20°C. This method has been successfully utilised to provide pharmacokinetic data throughout the clinical development of rosiglitazone. |
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