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Determination of Doxapram Hydrochloride in Rabbit Plasma by LC-MS-MS and Its Application

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Abstract

A sensitive and selective liquid chromatography tandem mass spectrometry (LC-MS-MS) method for determination of doxapram hydrochloride in rabbit plasma was developed. After addition of urapidil hydrochloride as internal standard (IS), protein precipitation by 10% trichloroacetic acid in methanol (w/v) was used as sample preparation. Chromatographic separation was achieved on a Zorbax SB-C18 (2.1 mm × 50 mm, 3.5 μm) column with acetonitrile–water as mobile phase with gradient elution. Electrospray ionization (ESI) source was applied and operated in positive ion mode; multiple reaction monitoring (MRM) mode was used for quantification using target fragment ions m/z 378.9 → 291.8 for doxapram hydrochloride and m/z 387.9 → 204.6 for the IS. Calibration plots were linear over the range of 2–1000 ng mL−1 for doxapram hydrochloride in plasma. Lower limit of quantitation (LLOQ) for doxapram hydrochloride was 2 ng mL−1. Mean recovery of doxapram hydrochloride from plasma was in the range 83.7–91.5%. RSD of intra-day and inter-day precision were less than 9%, respectively. This method is simple and sensitive enough to be used in pharmacokinetic research for determination of doxapram hydrochloride in rabbit plasma.

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Acknowledgment

This project was supported by the Analyzing and Testing Foundation of Zhejiang Province (2007F80010).

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Authors and Affiliations

  1. The First Affiliated Hospital of Wenzhou Medical College, 325000, Wenzhou, China

    Guanyang Lin & Lufeng Hu

  2. Function Experiment Teaching Center of Wenzhou Medical College, 325035, Wenzhou, China

    Jianshe Ma

  3. Analytical and Testing Center of Wenzhou Medical College, 325035, Wenzhou, China

    Xuebao Wang & Xianqin Wang

  4. Laboratory Animal Center of Wenzhou Medical College, 325035, Wenzhou, China

    Jiayin Zhu

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  1. Guanyang Lin
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  2. Jianshe Ma
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  3. Lufeng Hu
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  4. Xuebao Wang
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  5. Jiayin Zhu
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  6. Xianqin Wang
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Correspondence to Xianqin Wang.

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Lin, G., Ma, J., Hu, L. et al. Determination of Doxapram Hydrochloride in Rabbit Plasma by LC-MS-MS and Its Application. Chromatographia 73, 183–187 (2011). https://doi.org/10.1007/s10337-010-1815-3

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  • DOI: https://doi.org/10.1007/s10337-010-1815-3

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