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.

The primary vitamin in sea buckthorn berries is vitamin C containing values of approximately 400 mg/100 g. Processing effects were investigated during juice and concentrate production from sea buckthorn berries (Hippophaë rhamnoides) and storage stability of juices was determined for up to 7 d using berries and juices from 2 different growing areas. During industrial juice production the technological processing of the berries caused a loss of about 5% to 11% total ascorbic acid (TAA) in the generated juice. The production of the concentrated juice resulted in 50% depletion of TAA. Sea buckthorn berries and juice were stored at 6, 25, and 40 degrees C for up to 7 d to investigate the temperature effects on TAA during storage. Analysis of kinetic data suggested that the degradation follows a 1st-order model. The results of the experiments showed that storage of sea buckthorn juices for 7 d at cold temperature (6 degrees C) already resulted in a degradation of TAA of about 11% to 12%.

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A stable isotope dilution assay for quantitation of folate vitamers in sea buckthorn berries, 27 juice and concentrate using four-fold labeled folate isotopologues of the folate derivatives as 28 the internal standards was adopted using reversed phase liquid chromatography -tandem 29 Sea buckthorn products, Process and storage stability

Processing effects on the mineral content were investigated during juice production from sea buckthorn (Hippophaë rhamnoides L. ssp. rhamnoides, Elaeagnaceae) using berries from 2 different growing areas. The major and trace elements of sea buckthorn berries and juices were determined by atomic absorption spectroscopy (AAS)--(calcium, iron, magnesium, potassium, sodium) and inductively coupled plasma-mass spectrometry (ICP-MS)--(arsenic, boron, chromium, copper, manganese, molybdenum, nickel, selenium, zinc). Potassium is the most abundant major element in sea buckthorn berries and juices. The production process increased the potassium content in the juice by about 20%. Moreover, the processing of juice increased the value of manganese up to 32% compared to the content in berries. During industrial juice production, the technological steps caused a loss of about 53% to 77% of the chromium concentration, 50% of the copper content, 64% to 75% of the molybdenum amount, and up to 45% of the iron concentration in the final juice product. Consumption of sea buckthorn juice represents a beneficial source of chromium, copper, manganese, molybdenum, iron, and potassium for the achievement of the respective dietary requirements.