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.
Three major active components of the traditional Chinese medicinal herb Salvia miltiorrhiza Bunge, 3,4-dihydroxyphenyllactic 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; 50-100 mL, 20% ethanol, 20% acetic acid; and 100-200 mL, 30% ethanol, 30% acetic acid. The 3,4-dihydroxyphenyllactic 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.
Gu
M
M
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Box 353, Beijing, 100080, PR of China.
Su
Z-G
ZG
Janson
J-C
JC
D000327:Adsorption; D001547:Benzaldehydes / Q000302:isolation & purification*; D001572:Benzofurans / Q000302:isolation & purification*; D002396:Catechols / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry; D006860:Hydrogen Bonding; D007773:Lactates / Q000302:isolation & purification*; D027561:Salvia miltiorrhiza / Q000737:chemistry*; D012685:Sepharose; D053630:Depsides
D001547:Benzaldehydes; D001572:Benzofurans; D002396:Catechols; D004365:Drugs, Chinese Herbal; D007773:Lactates; D012685:Sepharose; C035055:3,4-dihydroxyphenyllactic acid; C005581:protocatechualdehyde; C076944:salvianolic acid B; D053630:Depsides