无患子总皂苷质控品的制备

无患子皂苷具有较好的洗涤功能,因其天然、安全、无毒,可开发成食品工业用清洗剂,而目前尚未有市售高纯度无患子总皂苷。以制备无患子总皂苷质控品为目标,通过乙醇提取、石油醚除杂、正丁醇萃取、AB-8大孔树脂、MCI凝胶柱分离纯化得无患子总皂苷,此无患子总皂苷再经液质联用（HPLC/MS）、Libermann-Buehard反应、溶血反应定性,香草醛-冰醋酸比色法定量。Libermann-Buehard反应颜色由黄-红-紫-蓝,有溶血性。HPLC/MS分析鉴定主要为齐墩果酸型三萜皂苷,分别为Sapindoside B（882）、Sapindoside A（750）、Mukurozi-saponin E1（924）、Sapindoside L（924）、Mukurozi-saponin G（966）、Sapindoside M（966）。以齐墩果酸为标准品,通过香草醛-冰醋酸显色,皂苷元在550nm有最大吸收,进一步判定无患子总皂苷以齐墩果酸型皂苷为主,且用35%盐酸高温高压水解无患子总皂苷2h,皂苷元得率最高,通过换算可知,无患子总皂苷的纯度能够达到95.2%,可作为无患子总皂苷质控品。 

Preparation of total sapindus-saponins for quality control

Sapindus saponin has wonderful washing fuction which is natural, environment-frendly and nontoxic. It can be developed into food industry cleaning agent. However, there is no pure sapindus saponin commercially available. With the goal of producing the total sapindus saponin for quality control, the method of ethanol extraction, butanol extraction, AB-8 macroporous resin and MCI Gel column separation was used to separate and purify sapindus saponin. The qualitative test was studied by means of Libermann-Buehard reaction and Hemolysis test. The quantitive test was made afterwards based on High Performance Liquid Chromatography/ Mass Spectrum and the method of vanillin-sulfuric acid. Libermann-Buehard reaction showed the color changes:yellow-red-purple-blue. Hemolysis test proved hemolytic. The components named Sapindoside B (882) , SapindosideA (750) , Mukurozi-saponinE1 (924) , Sapindoside L (924) , Mukurozi-saponin G (966) , Sapindoside M (966) were analysed by HPLC/MS, which were identified as Oleanolic acid saponins. Oleanolic acid was chosen as standard. Sapogenin of TSS after hydrolysis had a maximum absorption at 550nm with the method of vanillin-sulfuric acid, which was the same as Oleanolic acid in the condition of 35% hydrochloric acid, high temperature and pressure and time of 2h, the yield of sapogenin were the highest. The purity of TSS (Total Sapindus Saponin) was calculated to more than 95.2%. It could be further conclued that TSS was mainly oleanolic acid saponins and could be used for quality control as standard.