Simultaneous quantification of multiple licorice flavonoids in rat plasma
Flavonoids are important naturally occurring polyphenols with antioxidant properties. In this study, we report the development of a liquid chromatography tandem mass spectrometry (LC-MS/MS)-based method capable of simultaneously quantifying multiple active licorice flavonoids (including liquiritin apioside, liquiritin, liquiritigenin, isoliquiritin apioside, isoliquiritin, and isoliquiritigenin) in plasma. Electrospray ionization was used to efficiently generate precursor deprotonated molecules of all the analytes and the [M-H]− ions were used to produce characteristic product ions for MS/MS analysis. We found that inclusion of a very low concentration of HCOONH4 (0.01‰) in the LC mobile phase dramatically improved the detection limit for the tested flavonoids and decreased the interference by matrix effects, which have been referred to as “LC-electrolyte effects.” Liquid-liquid extraction with ethyl acetate was effective for isolation of all the analytes and resulted in the lowest matrix effects of several tested sample cleanup methods. This bioanalytical method showed good linearity between 0.32 ng/mL and 1 µg/mL analyte in 50-µL plasma samples. The accuracy and precision at different analyte concentrations varied from 85 to 110% and from 0.8 to 8.8%, respectively. Finally, we demonstrated the applicability of this method in a pilot pharmacokinetic study of rats receiving an oral dose of Xiaochaihu-tang, an important Chinese herbal remedy for chronic hepatitis. The use of a low concentration of HCOONH4 in the LC mobile phase could be used to improve LC-mass spectroscopy- or LC-MS/MS-based methods.
- U.S. FDA Guidance for Industry on Botanical Drug Products (http://www.fda.gov/cder/guidance/index.htm), 2004.
- Avery, M. J. (2003) Quantitative Characterization of Differential Ion Suppression on Liquid Chromatography/Atmospheric Pressure Ionization Mass Spectrometric Bioanalytical Methods. Rapid Commun. Mass Spectrom. 17: pp. 197-201 CrossRef
- Souverain, S., Rudaz, S., Veuthey, J. L. (2004) Matrix Effect in LC-ESI-MS and LC-APCI-MS with Off-line and On-line Extraction Procedures. J. Chromatogr. A 1058: pp. 61-66 CrossRef
- Bao, Y.-W., Li, C., Shen, H.-W., Nan, F.-J. (2004) Determination of Saikosaponin Derivatives in Radix bupleuri and in Pharmaceuticals of the Chinese Multiherb Remedy Xiaochaihu-tang Using Liquid Chromatographic Tandem Mass Spectrometry. Anal. Chem. 76: pp. 4208-4216 CrossRef
- Chen, P., Li, C., Liang, S.-P., Song, G.-Q., Sun, Y., Shi, Y.-H., Xu, S.-L., Zhang, J.-W., Sheng, S.-Q., Yang, Y.-M., Li, M. (2006) Characterization and Quantification of Eight Water-soluble Constituents in Tubers of Pinellia Ternata and in Tea Granules from the Chinese Multiherb Remedy Xiaochaihu-tang. J. Chromatogr. B 843: pp. 183-193 CrossRef
- Hsu, Y. L., Kuo, P. L., Lin, L. T., Lin, C. C. (2005) Isoliquiritigenin Inhibits Cell Proliferation and Induces Apoptosis in Human Hepatoma Cells. Planta Med. 71: pp. 130-134 CrossRef
- Homma, M., Oka, K., Taniguchi, C., Niitsuma, T., Hayashi, T. (1997) Systematic Analysis of Post-administrative Saiboku-to Urine by Liquid Chromatography to Determine Pharmacokinetics of Traditional Chinese Medicines. Biomed. Chromatogr. 11: pp. 125-131 CrossRef
- Matuszewski, B. K., Constanzer, M. L., Chavez-Eng, C. M. (2003) Strategies for the Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC-MS/MS. Anal. Chem. 75: pp. 3019-3030 CrossRef
- U.S. FDA Guidance for Industry on Bioanalytical Method Validation (http://www.fda.gov/cder/guidance/index.htm), 2001.
- Domon, B., Costello, C. E. (1988) A Systematic Nomenclature for Carbohydrate Fragmentation in FAB-MS/MS Spectra of Glycoconjugates. Glycoconjugate J. 5: pp. 397-409 CrossRef
- Temesi, D., Law, B. (1999) The Effect of LC Eluent Composition on MS Responses Using Electrospray Ionization. LC-GC 17: pp. 626-632
- Guo, B., Li, C., Wang, G.-J., Chen, L.-S. (2006) Rapid and Direct Measurement of Free Concentrations of Highly Protein-bound Fluoxetine and Its Metabolite Norfluoxetine in Plasma. Rapid Commun. Mass Spectrom. 20: pp. 39-47 CrossRef
- Simultaneous quantification of multiple licorice flavonoids in rat plasma
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