Purification of Allicin From Garlic Extract Using Semi-Preparative High Performance Liquid Chromatography

Background: Allicin has been reported to possess various biological properties. Commercial availability of allicin is limited because of its instability. Objectives: The aim of this study was to develop a semi-preparative high performance liquid chromatography (HPLC) method for purification of allicin from garlic extract. Materials and Methods: Aqueous garlic extract was prepared. Extract proteins were removed by methanol. The allicin peaks were separated using a semi-preparative C18 column and detected at 220 nm. Allicin fractions were collected and concentrated using a nonpolar solvent diethyl ether under vacuum at 33ºC. The content of purified allicin was quantitatively determined by analytical HPLC method. The biological activity was assayed using its antibacterial property compared to the standard allicin. Results: Semi-preparative HPLC yielded a distinct peak with a retention time of 12.28 minutes. The purity (≥ 95%) and biological activity (P > 0.05) of purified allicin were comparable to the standard allicin. Purified allicin produced same peak areas compared to the standard allicin in similar concentrations. The yield percentage of allicin in peak collection and concentrating steps were 100% and 73.34 ± 2%, respectively. Conclusions: This method could be feasibly used for purification of large quantities of allicin from garlic extract.