In view of the toxic inflammatory reaction induced by Euphorbia kansui roots, a traditional Chinese medicine used for the treatment of edema, ascites, and asthma, the 95% ethanol extract was found to have a significant stimulating effect on inflammatory cells. Bioassay-guided separation of the 95% ethanol extract from the roots of E. kansui led to the isolation of five diterpenoids whose structures were identified by (1)H, (13)C NMR spectroscopy and HR-ESI-MS as kansuinine B (1), kansuinine A (2), kansuiphorin C (3), 3-O-benzoyl-20-deoxyingenol (4), and 3-O-(2'E,4'Z-decadienoyl)-20-O-acetylingenol (5). The proinflammatory effect of compounds 1-5 was evaluated in vitro in models of inflammation using exoteric mice splenic lymphocytes (SPL) and rat peritoneal macrophages (PMphi). Compounds 1, 2, and 5 markedly promoted SPL proliferation and NO production by PMphi at concentrations from 0.78 to 12.50 microg/mL. Hence the three compounds are believed to be important proinflammatory components of the roots of E. kansui.
A high performance liquid chromatographic (HPLC) method with diode array detection (DAD) was established for simultaneous determination of seven main bioactive components in San-ao decoction and its series of formulae (San-ao decoction, Wu-ao decoction, Qi-ao decoction and Jia-wei San-ao decoction). Seven compounds were analyzed simultaneously with a XTerra C18 column (4.6 mm × 250 mm, 5 µm) using a linear gradient elution of a mobile phase containing acetonitrile (A) and a buffer solution (0.02 mol/L potassium dihydrogen phosphate and adjusted to pH 3 using phosphoric acid) (B); the flow rate was 1.0 mL/min. The sample was detected with DAD at 210, 254 and 360 nm and the column was maintained at 30 °C. All the compounds showed good linearity (r2 > 0.9984) in the tested concentration range. The precisions were evaluated by intra-day and inter-day tests, and relative standard deviation (R.S.D.) values within the range of 0.83%–2.53% and 0.64%–2.77% were reported, respectively. The recoveries of the quantified compounds were observed to cover a range from 95.34% and 104.82% with R.S.D. values less than 2.72%. The validated method was successfully applied for the simultaneous determination of seven main bioactive components including ephedrine (1), amygdalin (2), liquiritin (3), benzoic acid (4), isoliquiritin (5), formononetin (6) and glycyrrhizic acid (7) in San-ao decoction and its series of formulae. The results also showed a wide variation in the content of the identified active compounds in these samples, which could also be helpful to illustrate the drug interactions after some herbs combined in different formulations.
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