Development and validation of a high‐throughput and robust LC–MS/MS with electrospray ionization method for simultaneous quantitation of oseltamivir phosphate and its oseltamivir carboxylate metabolite in human plasma for pharmacokinetic studies |
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Authors: | Ravi Kanneti Dasandi Bhavesh Dharmesh Paramar Shivaprakash R Parloop A Bhatt |
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Institution: | 1. Department of Pharmacology, LM College of Pharmacy, Gujarat University, Ahmadabad 380015, India;2. Clinical Pharmacology and Pharmacokinetics, Dr Reddy's Laboratories Limited, Hyderabad 500072, India;3. Department of Bioanalytics and Clinical Research, Synchron Research Services Pvt. Ltd, The Chambers, 5th Floor, SG‐Highway, Ahmedabad 380054, India |
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Abstract: | A rapid, sensitive and rugged solid‐phase extraction ultraperformance liquid chromatography tandem mass spectrometry (LC‐MS/MS) method was developed for determination of oseltamivir phosphate (OP) and oseltamivir carboxylate (OC) in human plasma. The procedure for sample preparation includes a simple SPE extraction procedure coupled with a Chromatopack C18 column (50 × 3.0 mm, i.d., 3.0 µm) with isocratic elution at a flow‐rate of 0.600 mL /min and acyclovir was used as the internal standard. The analysis was performed on a triple‐quadrupole tandem mass spectrometer by multiple reaction monitoring mode via electrospray ionization. Using 500 µL plasma, the methods were validated over the concentration ranges 0.92–745.98 and 5.22–497.49 ng/mL for OP and OC, with a lower limit of quantification of 0.92and 5.22 ng/mL. The intra‐ and inter‐day precision and accuracy of the quality control samples were within 10.1%. The recovery was 68.72, 70.66 and 71.59% for OP, OC and IS, respectively. Total run time was only 1.0 min. The method was highly reproducible with excellent chromatography properties. Copyright © 2010 John Wiley & Sons, Ltd. |
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Keywords: | OP OC LC‐MS/MS Human plasma Electro Spray Ionization |
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