One-Step Purification of 3,4-Dihydroxyphenyllactic Acid, Salvianolic Acid B, and Protocatechualdehyde from Salvia miltiorrhiza Bunge by Isocratic Stepwise Hydrogen Bond Adsorption Chromatography on Cross-Linked 12% Agarose

M. Gu, Z.- G. Su, J.- C. Janson
2008 Journal of Chromatographic Science  
Three major active components of the traditional Chinese medicinal herb Salvia miltiorrhiza Bunge, 3,4dihydroxyphenyllactic acid, salvianolic acid B, and protocatechualdehyde, are separated and purified from a crude water extract in one step by isocratic hydrogen bond adsorption chromatography on cross-linked 12% agarose (Superose 12 HR 10/30). Separation is achieved by stepwise elution with mobile phases composed of mixtures of ethanol and acetic acid: 0-50 mL, 5% ethanol, 5% acetic acid;
more » ... % acetic acid; 50-100 mL, 20% ethanol, 20% acetic acid; and 100-200 mL, 30% ethanol, 30% acetic acid. The 3,4dihydroxyphenyllactic acid is obtained with a purity of 97.3% and with a recovery of 88.1%. The corresponding figures for protocatechualdehyde are a purity of 99.4% with a recovery of 90.7%, and for salvianolic acid B a purity of 90.4% with a recovery of 50.3%, respectively. At a sample load of 40 mg crude extract dissolved in 0.5 mL mobile phase (corresponding to a load of 1.6 mg/mL gel), a 3,4-dihydroxyphenyllactic acid purity of approximately 94% with a recovery of 80.2% is obtained.
doi:10.1093/chromsci/46.2.165 pmid:18366878 fatcat:z4kthq52mvgdlf3ltf3mp5obfu