title: Extraction and isolation of lithospermic acid B from Salvia miltiorrhiza Bunge using aqueous two-phase extraction followed by high-performance liquid chromatography.
abstract: A rapid and effective method integrating separation and purification of lithospermic acid B from Salvia miltiorrhiza Bunge was developed by combining an aqueous two-phase system extraction with preparative chromatography. An aqueous two-phase system of n-butyl alcohol/KH2 PO4 was chosen from seven systems. The influence of parameters including concentration of KH2 PO4 , n-butyl alcohol concentration, pH, and the ratio of an aqueous two-phase system to crude extract were investigated using a single factor design. Response surface methodology was subsequently used to find the optimal compositions of an aqueous two-phase system. Keeping a solvent-to-solid ratio of 10, the final optimized composition of an aqueous two-phase system was 39.1% w/w n-butyl alcohol and 22.6% w/w KH2 PO4 . Under these conditions a recovery yield of 99.8% and a high partition coefficient of 310.4 were obtained. In a pilot-scale experiment using optimized conditions, 18.79 g of lithospermic acid B with a purity of 70.5% and in a yield of 99.8% was separated from 0.5 kg of crude extract. Subsequently, 9.94 g lithospermic acid B with a purity of 99.3% and recovery yield of 70.3% was obtained with a preparative chromatographic process, and the two-step total recovery was 70.1%.
authors.affiliation: School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China. yongxueguo@163.com.
authors.forename: Yong Xue
authors.initials: YX
authors.lastname: Guo
authors.affiliation: School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China.
authors.forename: Chang Zhi
authors.initials: CZ
authors.lastname: Shi
authors.affiliation: School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Lv
authors.affiliation: School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, P. R. China.
authors.forename: Yue Yong
authors.initials: YY
authors.lastname: Zhang
mesh_terms: D001572:Benzofurans / Q000032:analysis / Q000302:isolation & purification*; D005591:Chemical Fractionation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D053630:Depsides / Q000032:analysis / Q000302:isolation & purification*; D004365:Drugs, Chinese Herbal / Q000032:analysis / Q000302:isolation & purification*; D027561:Salvia miltiorrhiza / Q000737:chemistry*
chemical_list: D001572:Benzofurans; D053630:Depsides; D004365:Drugs, Chinese Herbal; C076944:salvianolic acid B
keywords: Aqueous two-phase systems; Lithospermic acid B; Preparative chromatography; Response surface methodology