Application of preparative high-speed counter-current chromatography for the separation of flavonoids from the leaves of Byrsonima crassa Niedenzu (IK)

The methanolic extract of the leaves of the medicinal plant Byrsonima crassa (Malpighiaceae) contain flavonoids with antioxidant activity. They were separated in a preparative scale using high-speed counter-current chromatography. The optimum solvent system used was composed of a mixture of ethyl acetate-n-propanol-water (140:8:80 (v/v/v)) and led to a successful separation between monoglucosilated flavonoids (quercetin-3-O-alpha-L-arabinoside, quercetin-3-O-beta-D-galactoside) and the biflavonoid amentoflavone in only 3.5 h. The purities of quercetin-3-O-alpha-L-arabinoside (95 mg), quercetin-3-O-beta-D-galactoside (16 mg) and the biflavonoid amentoflavone (114 mg) were all isolated at purity over 95%. Identification was performed by 1H NMR, 13C NMR and UV analyses.     

加速溶剂萃取分离-液相色谱法测定五灵脂药材中3种成分的含量

取五灵脂药材(1.0g)与海沙3.0g混匀,置于加速溶剂萃取(ASE)仪中,加入萃取溶剂[乙醇-水(75+25)]静态萃取循环3次,每次10min,萃取温度为100℃,压力为15 MPa。萃取液蒸缩至3mL,加C2H5OH-H2O(75+25)至5.0mL,过滤后供液相色谱分析。在色谱分离中,以Diamosil ODS C18色谱柱为固定相,用(A)甲醇和(B)乙酸(0.5+99.5)溶液以不同比例的混合液作为流动相进行梯度淋洗。在波长360nm处进行检测。药材中的芦丁、山奈酚和穗花杉双黄酮的质量浓度与相应的峰面积之间呈线性关系,3种组分的检出限(3S/N)在0.3~0.6mg·kg-1之间。在3个浓度水平条件下用标准加入法进行回收试验,测得回收率在80.0%~95.0%之间,测定值的相对标准偏差(n=5)在3.2%~9.6%之间。