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KMID : 0861020100250030065
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Korea Journal of Herbology
2010 Volume.25 No. 3 p.65 ~ p.71
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Simultaneous Determination of Seven Compounds by HPLC-PDA and Cytotoxicity of Samchulkunbi-tang
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Seo Chang-Seob
Lee Mi-Young
Kim Jung-Hoon
Lee Jin-Ah
Shin Hyeun-Kyoo
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Abstract
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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 column operating at , 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 >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 /mL concentrations.
Conclusions：The established method will be helpful to improve quality control and in vitro efficacy study of SKT.
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KEYWORD
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HPLC-PAD, samchulkunbi-tang (SKT), simultaneous determination, cytotoxicity, BEAS-2B cell
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