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http://dx.doi.org/10.6116/kjh.2010.25.3.065

Simultaneous Determination of Seven Compounds by HPLC-PDA and Cytotoxicity of Samchulkunbi-tang  

Seo, Chang-Seob (Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine)
Lee, Mee-Young (Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine)
Kim, Jung-Hoon (Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine)
Lee, Jin-Ah (Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine)
Shin, Hyeun-Kyoo (Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine)
Publication Information
The Korea Journal of Herbology / v.25, no.3, 2010 , pp. 65-71 More about this Journal
Abstract
Objectives：To develop and validate HPLC-PDA methods for simultaneous determination of seven constituents in Samchulkunbi-tang (SKT). Additionally, we investigated the cytotoxicity against BEAS-2B cell line and splenocytes of SKT. Methods：Reverse-phase chromatography using a Gemini $C_{18}$ column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 230, 254 and 280 nm, were used for quantification of the three marker components of SKT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. The cytotoxicity of SKT were measured by the CCK-8 assay method. Results：Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD) values (%) for intra- and inter-day precision were less than 6.0%. The recovery rate of each compound was in the range of 86.89-109.78%, with an RSD less than 4.0%. The contents of seven compounds in SKT were 1.39-6.84 mg/g. SKT had no cytotoxicity effect at 50-200 ${\mu}g$/mL concentrations. Conclusions：The established method will be helpful to improve quality control and in vitro efficacy study of SKT.
Keywords
HPLC-PAD; samchulkunbi-tang (SKT); simultaneous determination; cytotoxicity; BEAS-2B cell;
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