An improved HPLC/MS method for the analysis of escins in Common Horse-chestnut (Aesculus hippocastanum)

The deciduous tree, Aesculus hippocastanum L. [Sapindaceae] known as the Common Horse-chestnut is native to a small area in the mountains of the Balkans, but is now widely cultivated in temperate regions around the world. The extract from the seed finds widespread use in the treatment of chronic venous insufficiency, edema, and sprains. The components of the extract primarily responsible for the pharmacological effect are a group of saponin compounds collectively known as escins, and is comprised of over thirty glycosidic triterpene compounds derived from the aglycone protoaescigenin or barringtogenol C. HPTLC is the current analysis method for total escins in raw material or products. Through the use of HPLC/MS, an analysis method has been developed which also allows for the determination of total escins. By selecting specific fragments due to ionization the percent total escins can be quantified reproducibly. A number of products on the market in addition to raw material were tested and the values were compared to specifications when available. In most cases, products achieved label claim, but not all. Extract procedures can also cause variation in products. This method is able to determine if the product is representative of the raw material extract, or if it has been changed due to extreme extract procedures.