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Analysis of Biologically Active Constituents in Centella asiatica by Microwave-Assisted Extraction Combined with LC–MS

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Abstract

Centella asiatica (L.) Urban is a traditional herbal medicine used in Asiatic countries, and is commonly used to treat various wounds, leprosy, tuberculosis and lupus diseases. In this work, a new method based on microwave assisted extraction followed by liquid chromatography–diode array detection–electrospray ionization multistage tandem mass spectrometry analysis has been developed for the identification and quantification of biologically active constituents in C. asiatica, including asiatic acid, asiaticoside and madecassoside. The separation was performed on an Agilent Eclipse C18 column (150 mm × 4.6 mm i.d., 5 μm) with gradient elution of acetonitrile and 0.1% aqueous acetic acid within 50 min. Detection was performed at 205 nm. The calibration curves showed good linearity (r 2 > 0.9992). The limits of detection ranged from 1.2 to 1.6 μg mL−1. The intra- and inter-day precision was less than 3% and the recovery of the assay was in the range of 95.4–106.8%. The method was successfully applied to the quantification of the three constituents in different samples of C. asiatica. The results indicated that the developed method could be considered to be a simple, rapid and reliable method for the quality evaluation of C. asiatica. The samples were also analyzed on a liquid chromatography–electrospray ionization–time-of-flight mass spectrometry system to confirm the identification results.

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Acknowledgments

Financial support from the Natural Science Foundation of Zhejiang Province (Y2080331), and Wenzhou Municipal Science and Technology Bureau (H20080052) is gratefully acknowledged.

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Authors and Affiliations

  1. College of Chemistry and Materials Science, Wenzhou University, 325027, Wenzhou, People’s Republic of China

    Yan Shen, Aili Liu & Mingde Ye

  2. College of Food Science and Biotechnology Engineering, Zhejiang Gongshang University, 310035, Hangzhou, People’s Republic of China

    Li Wang

  3. Qingdao Key Lab of Analytical Technology Development and Standardization of Chinese Medicines, First Institute Oceanography of SOA, 266061, Qingdao, People’s Republic of China

    Junhui Chen & Xiao-ru Wang

  4. Wenzhou Entry-Exit Inspection and Quarantine Bureau of PRC, 325027, Wenzhou, People’s Republic of China

    Chao Han

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  1. Yan Shen
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  2. Aili Liu
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  3. Mingde Ye
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  4. Li Wang
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  5. Junhui Chen
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  6. Xiao-ru Wang
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Correspondence to Chao Han.

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Shen, Y., Liu, A., Ye, M. et al. Analysis of Biologically Active Constituents in Centella asiatica by Microwave-Assisted Extraction Combined with LC–MS. Chroma 70, 431–438 (2009). https://doi.org/10.1365/s10337-009-1152-6

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  • DOI: https://doi.org/10.1365/s10337-009-1152-6

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