Li Ke-Ke, Xu Fei, Gong Xiao-Jie
College of Medical, Dalian University, Dalian 116622, China.
Molecules. 2016 Mar 9;21(3):315. doi: 10.3390/molecules21030315.
In this paper, the isolation, purification and quantification of ginsenoside F₅ and F₃ isomeric compounds from crude extracts of flower buds of Panax ginseng (CEFBPG) was investigated by reversed-phase high-performance liquid chromatography (RP-HPLC) for the first time. The satisfied separation at analytical scale was achieved using a Zorbax Eclipse XDB C-18 column with a ternary mobile phase of acetonitrile-water-phosphoric acid (28:71:1) at a flow rate of 1.0 mL/min within 40 min. UV detection was set at 203 nm. Ginsenoside F₅ and F₃ was 4.21 mg and 5.13 mg in 1 g flower buds of P. ginseng (FBPG), respectively. The preparation of ginsenoside F₅ and F₃ at semi-preparative scale was performed by using a Daisogel C-18 column and gradient elution system of acetonitrile-water (32:68 → 28:72) at a flow rate of 10 mL/min with a sample load of 20-30 mg, and yielded ginsenosides in purity of more than 96%. Their structures were characterized by NMR and high resolution electrospray ionization mass spectrometry (HRESIMS). All the method validations showed acceptable limits. The results indicate a new source to obtain ginsenoside F₅ and F₃, and show that the method developed here appears to be reliable for simultaneously preparing them from CEFBPG.
本文首次采用反相高效液相色谱法(RP-HPLC)对人参花蕾粗提物(CEFBPG)中人参皂苷F₅和F₃异构体化合物进行了分离、纯化和定量研究。使用Zorbax Eclipse XDB C-18色谱柱,以乙腈-水-磷酸(28:71:1)的三元流动相,流速为1.0 mL/min,在40分钟内实现了分析规模的满意分离。紫外检测波长设定为203 nm。人参花蕾(FBPG)中1 g含人参皂苷F₅ 4.21 mg和人参皂苷F₃ 5.13 mg。采用Daisogel C-18色谱柱和乙腈-水(32:68 → 28:72)梯度洗脱系统,流速为10 mL/min,进样量为20 - 30 mg,进行半制备规模的人参皂苷F₅和F₃制备,得到纯度超过96%的人参皂苷。通过核磁共振(NMR)和高分辨电喷雾电离质谱(HRESIMS)对其结构进行了表征。所有方法验证均显示出可接受的限度。结果表明获得人参皂苷F₅和F₃的新来源,并表明本文建立的方法对于从CEFBPG中同时制备它们似乎是可靠的。