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Analysis of Linarin and Its Metabolites in Rat Urine by LC–MS/MS

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Abstract

Liquid chromatography–ion trap mass spectrometry was employed to investigate the metabolism of linarin in rats. Identification and structural elucidation of the metabolites were performed by comparing the differences in molecular masses, retention times, and full scan MSn spectra between linarin and its metabolites. Six metabolites (acacetin, apigenin, acacetin glucuronide, apigenin glucuronide, acacetin sulfate, apigenin sulfate) were detected in rat urine after oral administration of linarin at the dose of 50 mg kg−1. Furthermore, a selective and sensitive liquid chromatography–triple quadruple mass spectrometry assay was developed and validated for the simultaneous determination of linarin and acacetin (the major metabolite of linarin) in rat urine. Chromatographic separation was carried out on a C18 column, and mass spectrometric detection was performed using a triple-quadrupole mass spectrometer coupled with an electrospray ionization source in the positive ion mode. Quantitation of linarin and acacetin was performed using selected reaction monitoring of precursor–product ion transitions at m/z 593 → 285 for linarin, 285 → 242 for acacetin, and 303 → 153 for hesperitin (internal standard), respectively. The assay exhibited good linearity (r > 0.9900) for both linarin and acacetin. The intra- and inter-day precisions were <13.4 % and the accuracy was between −8.1 and 3.1 %. The method was successfully applied to the urinary excretion study of linarin in rats after oral administration of linarin.

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Acknowledgments

This study was supported by a Key Project for Drug Innovation grant from the Ministry of Science and Technology of China (2013zx09301305). The authors would like to thank Liang Chen for his excellent technical supports.

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Authors and Affiliations

  1. School of Pharmacy, Shenyang Pharmaceutical University, 103 Wenhua Road, PO Box 54, Shenyang, 110016, People’s Republic of China

    Xinchi Feng, Youping Liu, Xin Wang & Xin Di

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  1. Xinchi Feng
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  2. Youping Liu
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  3. Xin Wang
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  4. Xin Di
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Correspondence to Xin Di.

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Feng, X., Liu, Y., Wang, X. et al. Analysis of Linarin and Its Metabolites in Rat Urine by LC–MS/MS. Chromatographia 77, 571–579 (2014). https://doi.org/10.1007/s10337-014-2641-9

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  • DOI: https://doi.org/10.1007/s10337-014-2641-9

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