Systematic Separation and Purification of Iridoid Glycosides and Crocetin Derivatives from Gardenia jasminoides Ellis by High-speed Counter-current Chromatography

IntroductionIridoid glycosides and crocetin derivatives are the main bioactive components of Gardenia. The processes of separation of these compounds reported in much of the literature are tedious, time consuming and require multiple chromatographic steps, which results in lower recovery and higher costs. ObjectiveTo develop a high-speed counter-current chromatography (HSCCC) method for the systematic separation and purification of iridoid glycosides and crocetin derivatives on a preparative scale from Gardenia. MethodsAfter fractionation using HPD100 column chromatography, n-butanol:ethanol:water (10:1:10, v/v) was selected to purify gardenoside, 6-hydroxy geniposide and geniposidic acid from fraction A; ethyl acetate:n-butanol:water (2:1.5:3, v/v) was used to isolate geniposide from fraction B; crocin-1, crocin-2, crocin-3 and crocin-4 were purified by hexane:ethyl acetate:n-butanol:water (1:2:1:5, v/v) from fraction C. The head-to-tail elution mode was used with a flow rate of 8.0mL/min and a rotary speed of 600rpm. ResultsAfter HSCCC isolation, 151.1mg of gardenoside, 52.2mg of 6-hydroxy geniposide and 24.5mg of geniposidic acid were obtained from 800mg of fraction A; 587.2mg of geniposide was obtained from 800mg of Fraction B; 246.2mg of crocin-1, 34.2mg of crocin-2, 24.4mg of crocin-3 and 24.7mg of crocin-4 were obtained from 1000mg of fraction C. Their purities were found by UPLC analysis to be 91.7%, 93.4%, 92.5%, 98.2%, 94.1%, 96.3%, 94.1% and 98.9% respectively. ConclusionThe present results demonstrates that the main iridoid glycosides and crocetin derivatives in Gardenia can be obtained efficiently from extracts using HSCCC. Copyright (c) 2015 John Wiley & Sons, Ltd. This is a first paper reporting a HSCCC method for systematic isolation and purification of main crocetin derivatives and iridoid glycosides from Gardenia jasminoides Ellis. 151.1 mg of gardenoside, 52.2 mg of 6-hydroxy geniposide, 24.5 mg of geniposidic acid, 587.2 mg of geniposide, 246.2 mg of crocin-1, 34.2 mg of crocin-2, 24.4 mg of crocin-3 and 24.7 mg of crocin-4 were obtained from three fractions pretreated from gardenia extract. The main iridoid glycosides and crocetin derivatives in gardenia could be efficiently obtained using HSCCC from the extract of gardenia.