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KMID : 0363620110320030025
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Journal of Korean Oriental Medicine
2011 Volume.32 No. 3 p.25 ~ p.34
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Evaluation of the Immune-Stimulating Activity of Samul-tang, a Traditional Korean Herbal Medicine, Standardized by HPLC-PDA
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Seo Chang-Seob
Ha Hye-Kyung
Jung Da-Young
Lee Ho-Young
Shin Hyeun-Kyoo
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Abstract
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Objectives: We performed simultaneous determination of five constituents by HPLC in Samul-tang (SMT). Additionally, we investigated the immune-stimulatory potential of SMT on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice.
Methods: Reverse-phase chromatography using a Gemini C18 column operating at , and photodiode array (PDA) detection at 190-400 nm, were used for quantification of the five components of SMT. 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. C57BL/6 mice were immunized intraperitoneally with OVA/alum () on days 1, 8, and 15. The extract of SMT (1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma.
Results: Calibration curves were acquired with >0.9999, and the relative standard deviation (RSD, %) for intra- and inter-day precision were both less than 3.5%. The recovery was in the range of 95.69-115.12%, with an RSD less than 6.0%. The contents of five components in SMT were 1.08-15.30 mg/g. SMT significantly enhanced Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). Also, SMT significantly enhanced OVAspecific IgG, IgG1 and total IgM levels in plasma compared with the OVA-immunized group.
Conclusions: The established method will be applied for the quantification of major components and immunestimulating activity in OVA-immunized mouse model of SMT.
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KEYWORD
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Herbal medicine, HPLC-PDA, Samul-tang, simultaneous determination, immune-stimulating activity
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