Preparative Isolation and Purification of Flavonoids and Protocatechuic Acid from Sea Buckthorn Juice Concentrate (Hippophaë rhamnoides L. ssp. rhamnoides) by High-Speed Counter-Current Chromatography

Abstract

This is a postprint of an article published in Gutzeit, D., Wray, V., Winterhalter, P., Jerz, G. Preparative isolation and purification of flavonoids and protocatechuic acid from sea buckthorn juice concentrate (Hippophae? rhamnoides L. ssp. rhamnoides) by high-speed counter-current chromatography (2007) Chromatographia, 65 (1-2), pp. 1-7. Abstract High-speed counter-current chromatography (HSCCC) -a support free all liquid-liquid chromatography technique -has been successfully used for the preparative isolation of isorhamnetin 3-O-ß-D-glucoside, isorhamnetin 3-O-ß-rutinoside, quercetin 3-O-ß-Dglucoside, syringetin 3-O-ß-D-glucoside and protocatechuic acid from sea buckthorn juice concentrate (Hippophaë rhamnoides L. ssp. rhamnoides, Elaeagnaceae). The preparative HSCCC instrument was a multilayer coil planet centrifuge equipped with three preparative coils. Separation was performed with a two phase solvent system (nhexane -n-butanol -water, 1:1:2 v/v/v) in 'head-to-tail' mode. Each injection of 4.1 g crude ethyl acetate extract yielded isorhamnetin 3-O-ß-D-glucoside (95 mg), isorhamnetin 3-O-ß-rutinoside (10 mg), quercetin 3-O-ß-D-glucoside (5 mg), and protocatechuic acid (34 mg) with purities > 98 %. The flavonoid syringetin 3-O-ß-Dglucoside (2 mg) was a novel compound for Hippophaë rhamnoides. Chemical structures of all compounds were determined by HPLC-ESI-MS-MS, 1D-NMR ( 1 H, 1 13 C, DEPT 135) spectroscopy and for elucidation of glycosidic linkages 2D-NMR (HMBC) spectroscopy was used.

Genesis and development of DPPH method of antioxidant assay

“…HSCCC is a liquid-liquid chromatographic technique with no solid support matrix; therefore eliminates the irreversible adsorption of samples (Yoichiro 1981). This method has been successfully used to separate and isolate many natural products (David et al 2007;Gutzeit et al 2007.…”

Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

“…31 A substância quercetina foi utilizada como padrão referência e o cálculo da CI 50 foi obtido através da seguinte fórmula:…”

“…Essa hipótese é fundamentada na identificação dos mesmos em espécies do gênero, como em B. purpurea e B. blakeana. [50][51][52][53] Os íons em m/z 289 e 577 em GBp1-F podem estar relacionados à presença da epicatequina e duas unidades da mesma, chamada epicatequina-epicatequina, também já identificada anteriormente em espécies de Bauhinia, como B. championii Benth. e B. aurea.…”

Preparative isolation and purification of alkannin/shikonin derivatives from natural products by high‐speed counter‐current chromatography

“…Artifact formation is also eliminated, since it used no solid support. HSCCC has been used for analysis, separation and purification of bioactive compounds from natural products, such as alkaloids, hydroxyanthraquinones, flavonoids and saponins (Shibusawa et al, 2000;Liu et al, 2004;Aman et al, 2005;Chu et al, 2005;Li and Chen, 2005;Zhou et al, 2006;Gutzeit et al, 2007;Yao et al, 2007). This technique presents maximum capacity with an excellent sample recovery and high efficiency, and can be employed for preparative-scale separations in a straightforward manner (Tong et al, 2006).…”