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A new strategy for quality control and qualitative analysis of Yinhuang preparations by HPLC-DAD-MS/MS

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Abstract

A combinative method using fingerprint analysis (FA) and multi-ingredients quantification (MIQ) was developed and validated for the quality control of Yinhuang (YH) preparations including granule, capsule, and lozenge by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). Common peaks with or without standard references in FA were confirmed or identified by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The chromatographic separations were achieved on a Sepax GP-C18 column (250 mm × 4.6 mm i.d., 5 μm) with a gradient elution using a mixture of 0.1 % formic acid methanol solution and 0.1 % formic acid water solution. In quantitative analysis, nine bioactive constituents (chlorogenic acid, caffeic acid, luteoloside, baicalin, luteolin, wogonoside, baicalein, wogonin, and oroxylin A) were simultaneously determined. The detection wavelength was set at 275 nm, 320 nm, and 350 nm according to the absorption properties of the nine quantified compounds. The linearity, recovery, intraday and interday precision, accuracy, limit of detection (LOD) and quantification (LOQ), repeatability and stability were all tested and good results were obtained. In the FA, 320 nm was selected. The correlation coefficients of similarity were determined on the basis of the relative retention time (RRT) and relative peak area (RPA) of 20 common peaks in chromatographic fingerprints. Results indicated that both the RRT and RPA of 20 common peaks shared a close similarity. From the 20 common peaks, 18 compounds, including the nine quantified compounds, were identified or tentatively characterized by comparing their retention times, UV spectra, and MS spectra with those of standard compounds or literature data. The study not only presents a powerful and reliable analytical tool for the quality control of YH preparations, but also provides the chemical evidence for revealing the material basis of their therapeutic effects.

The HPLC-DAD and LC-ESI-MS/MS chromatogram of Yinhuang granule

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Acknowledgments

The authors acknowledge support for this work by a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the Xuzhou Natural Science Foundation of China (no. XX10A052), and Jiangsu Aosaikang Foundation for Clinical Pharmacy (201012).

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Authors and Affiliations

  1. Key Laboratory of New Drug & Clinical Application, Xuzhou Medical College, Xuzhou, Jiangsu, 221004, China

    Hui Chen, Xu Chen, Jing Wu, Dao-quan Tang, Qian Du, Xiao-xing Yin & Dong-zhi Yang

  2. Department of Pharmacy, Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221002, China

    Qiang Han

  3. Department of Pharmaceutical Analysis, Xuzhou Medical College, Xuzhou, Jiangsu, 221004, China

    Dao-quan Tang & Dong-zhi Yang

Authors
  1. Hui Chen
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  2. Xu Chen
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  3. Qiang Han
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  4. Jing Wu
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  5. Dao-quan Tang
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  6. Qian Du
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  7. Xiao-xing Yin
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  8. Dong-zhi Yang
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Corresponding author

Correspondence to Dao-quan Tang.

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Hui Chen and Xu Chen contributed equally to this work.

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Chen, H., Chen, X., Han, Q. et al. A new strategy for quality control and qualitative analysis of Yinhuang preparations by HPLC-DAD-MS/MS. Anal Bioanal Chem 404, 1851–1865 (2012). https://doi.org/10.1007/s00216-012-6281-3

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  • DOI: https://doi.org/10.1007/s00216-012-6281-3

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