Development and validation of a rapid and high‐sensitivity liquid chromatography–tandem mass spectrometry assay for the determination of neostigmine in small‐volume beagle dog plasma and its application to a pharmacokinetic study |
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Authors: | Di Cao Wenxue Li Xin Zhao Xiaolan Ye Fanlu Sun Jinying Li Fenyun Song Guorong Fan |
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Affiliation: | 1. Department of Pharmacy, Guangdong Pharmaceutical University, Guangdong, People's Republic of China;2. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai, People's Republic of China;3. Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Shanghai, People's Republic of China;4. College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian, People's Republic of China;5. Xuhui Central Hospital of Shanghai, Shanghai, People's Republic of China |
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Abstract: | A simple, rapid and high sensitive liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method for the determination of neostigmine in small‐volume beagle dog plasma was developed to assess the plasma pharmacokinetics of neostigmine. After protein precipitation in a Sirocco 96‐well filtration plate, the filtrate was directly injected into the LC‐MS/MS system. The analytes were separated on a Hanbon Hedera CN column (100 × 4.6 mm, 5 µm) with a mobile phase composed of methanol–water (60:40, v/v) and the water containing 0.01% formic acid at a flow rate of 0.6mL/min, with a split ratio of 1:1 flowing 300 μL into the mass spectrometer. The run time was 3 min. Detection was accomplished by electrospray ionization source in multiple reactions monitoring mode with the precursor‐to‐product ion transitions m/z 223.0 → 72.0 and 306.0 → 140.0 for neostigmine and anisodamine (internal standard), respectively. The method was sensitive with a lower limit of quantitation of 0.1 ng/mL, and good linearity in the range 0.1–100ng/mL for neostigmine (r ≥ 0.998). All the validation data, such as accuracy, intra‐run and inter‐run precision, were within the required limits. The method was successfully applied to pharmacokinetic study of neostigmine methylsulfate injection in beagle dogs. Copyright © 2013 John Wiley & Sons, Ltd. |
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Keywords: | neostigmine LC‐MS/MS Sirocco 96‐well filtration plates pharmacokinetic study Beagle dogs |
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