Taking application of some isolation and purification technologies, including crushing, solvent extraction, preliminary solvent isolation, column chromatographies over silica gel and Sephadex LH-20 gel and preparative HPLC, 8 compounds were obtained from the seeds of Jufeng grape sourced from market. Their structures were identified by spectroscopic methods and comparison with literature values as Catechin (1), Epicatechin (2), quercetin (3), ethylgallate (4), rel-(2S, 3R) -2-(4-hydroxy-3-methoxyphenyl) -3- (hydroxymethyl)-5-(3-hydroxypropyl)-2,3-dihydrobenzofuran-7-ol (5), rel-(2α, 3β)-7-O-methylcedrusin (6), rel-(1R,2S)-1-(4-hydroxy-3-methoxyphenyl) -2-(4-(3-hydroxypropyl) -2-methoxyphenoxy) propane-1,3-diol (7), and (+) -isolariciresinol (8), respectively. Compounds 5-8 were serial lignans isolated from the seeds of grape for the first time. Structurally, 5 and 6 belong in benzofuran-8,3'-neolignans, 7 in 8,4'-oxyneolignan, and 8 in 8,8' :2,7'-cyclolignan. According to in vitro activity evaluation conducted in cell model, compound 6 showed significant anti-oxidative ability, with the activity of RAW264. 7 cell superoxide dismutase being raised evidently in the test as compared with the positive anti-oxidative agents, compounds 1 and 2.
Antioxidants ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Plant Extracts ; chemistry ; isolation & purification ; Seeds ; chemistry ; Vitis ; chemistry

This article studied the chemical constituents from the aerial part of Vitis thunbergii var. taiwaniana. The 60% ethanol extract was eluted with 95% ethanol though HP-20 macroporous adsorption resin column. 12 compounds, including (1) betulinic acid, (2)2, 2, 2'-bis (4-hydroxyphenyl) propane bis (2, 3-epoxypropyl) ether, (3) eriodictyol, (4) trans-ε-viniferin, (5) (+)-cis-ε-viniferin, (6) kobophenol A, (7) ampelopsin A, (8) nepalensinol B, (9) cis-miyabenol C, (10) cis-vitisin B, (11) cis-gnetin H and (12) (+)-hopeaphenol, were separated by using normal phase silica gel, ODS, Sephdadex LH-20 column chromatographies and semi-preparative or preparative HPLC. Compounds 2, 5, 6, 8, 9, 10, 11 were separated from the genus Vitis for the first time and compounds 3, 7, 12 were separated from Vitis thunbergii var. taiwaniana for the first time. At a concentration of 50 μmol · L(-1), compound 6, 7 and 11 showed strong cytotoxicity against MCF-7 cell lines with the inhibition rate of 66.58%, 57.16%, 52.84%, respectively.
Antineoplastic Agents, Phytogenic ; pharmacology ; Humans ; MCF-7 Cells ; Plant Extracts ; analysis ; pharmacology ; Vitis ; chemistry

The instability of secondary metabolite production is a ubiquitous problem in plant cell culture. In order to understand the instability in plant cell culture, we investigated anthocyanin accumulation in suspension cultures of Vitis vinifera, as a model system, in our laboratory. Not only the anthocyanin contents but also its composition exhibited instability along with the long-term subculture. New methods were developed to indicate the instability of plant cell culture. Both the definition of instability coefficient (delta) and the application of factor scores were the first time in this field. To examine the effects of culture conditions on instability of anthocyanin biosynthesis, different subculture cycles and inoculum sizes had been investigated. Subculture cycle and inoculum size were both environmental cues driving the instability. Compared with subculture cycle, inoculum size was more effective in working on the instability of anthocyanin accumulation. Among all the conditions investigated in our study, (6.5 d, 2.00 g), (7 d, 2.00 g), (7.5 d, 2.00 g), (7 d, 1.60 g) and (7 d, 2.40 g), the condition of 7 d-subculture cycle together with 1.60 g-inoculum size was the best one to keep the stable production of anthocyanins.
Anthocyanins ; biosynthesis ; chemistry ; Culture Techniques ; methods ; Time Factors ; Vitis ; growth & development ; metabolism

The current situation of plant extract in domestic and international market was analyzed in the paper. The quality control of 20 plant extracts which have reasonably good sales in USA market was compared and analyzed. The analysis of the quality control of six plant extracts indicated that there were two main reasons leading to the varied quality specifications among different suppliers. One reason was that the plant species utilized by different companies were different. The other reason was that the extraction processes were different among different production plants. Comparing with the significant international suppliers of plant extracts, the product quality of Chinese companies were not satisfactory. It was suggested that chromatography and chromatographic fingerprint techniques should be applied to improve the quality control standard of plant extract in our country.
Anthocyanins ; isolation & purification ; China ; Chromatography ; methods ; Plant Extracts ; isolation & purification ; standards ; Plants, Medicinal ; chemistry ; Proanthocyanidins ; isolation & purification ; Quality Control ; Species Specificity ; Vaccinium ; chemistry ; Vitis ; chemistry

OBJECTIVETo optimize the the ultrasound-assisted subcritical water extraction (USWE) parameters of proanthocyanidins from defatted grape seed, study antioxidant activity of proanthocyanidins and compare the effects of USWE and other extraction techniques.

METHODThe 2 L equipment of USWE was designed and used to extract the proanthocyanidins. The factors including extraction temperature, extraction time and extraction pressure were studied. The best extraction condition was found through the response surface design. Antioxidant activity of proanthocyanidins was studied by its DPPH free radical and NaNO2 scavenging action.

RESULTThe USWE parameters were extraction temperature 145 degrees C, extraction time 18 min, extraction pressure 14 MPa and the extraction yield (EY) was 4.05% under this extraction condition. The proanthocyanidins extracted under this optimized extraction condition had better scavenging action on DPPH free radical and NaNO2. As compared with the conventional soxhlet's extraction and heat reflux extraction, the USWE cost less extraction time, and possessed high efficiency and so on.

CONCLUSIONThe extraction technology of USWE is highly feasible to extract proanthocyanidins from defatted grape seed.

Analysis of Variance ; Chemical Fractionation ; methods ; Free Radical Scavengers ; isolation & purification ; pharmacology ; Pressure ; Proanthocyanidins ; isolation & purification ; pharmacology ; Seeds ; chemistry ; Temperature ; Time Factors ; Ultrasonics ; Vitis ; chemistry ; Water ; chemistry

This paper reiviewed the current situation of quality control of grape seed extract in domestic and international market. Considering the fact that there is no national or industrial technical specifications established for the extract product, the authors suggested that two sets of quality specifications should be established for the grape seed extract. The two sets of specifications are: the high purity grape seed extract should contain polyphenol NLT 95%, monomer NLT 10%; and the grape seed extract with ordinary quality should have a procyanidolic value NLT 95, and monomer NLT 6%.
Antioxidants ; chemistry ; isolation & purification ; standards ; Biflavonoids ; analysis ; isolation & purification ; Catechin ; analysis ; isolation & purification ; Flavonoids ; analysis ; isolation & purification ; Phenols ; analysis ; isolation & purification ; Plant Extracts ; chemistry ; isolation & purification ; standards ; Polyphenols ; Proanthocyanidins ; analysis ; isolation & purification ; Quality Control ; Seeds ; chemistry ; Vitis ; chemistry

We analyzed the best light intensity for callus induction and maintenance in Vitis vinifera and explored the mechanism of grape callus browning. Tender stem segments of grape cultivar "gold finger" were used to study the effects of different light intensities (0, 500, 1 000, 1 500, 2 000, 2 500, 3 000 and 4 000 Lx) on the induction rate, browning rate and associated enzyme activity and gene expression during Vitis vinifera callus formation. The callus induction rate under 0, 500, 1 000 and 1 500 Lx was more than 92%, significantly higher than in other treatments (P < 0.05). A lower browning rate and better callus growth were also observed during subculture under 1 000 and 1 500 Lx treatments. We found that chlorogenic acid, caffeic acid, p-hydroxybenzoic acid and coumaric acid contents were correlated with the browning rate of callus, among which chlorogenic acid content was positively correlated with the browning rate (P < 0.05). Peroxidase (POD) and polyphenol oxidase (PPO) activities were negatively correlated with the browning rate of callus (P < 0.01). The POD, PPO and phenylalanine ammonialyase (PAL) expression levels were positively correlated with the browning rate at P < 0.05 or P < 0.01. An appropriate light intensity for the tissue culture of Vitis vinifera was 1 000-1 500 Lx, higher or lower light intensities significantly impaired normal callus growth.
Caffeic Acids ; chemistry ; Catechol Oxidase ; chemistry ; Culture Media ; chemistry ; Gene Expression Regulation, Plant ; Light ; Peroxidase ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Stems ; enzymology ; radiation effects ; Tissue Culture Techniques ; Vitis ; enzymology ; radiation effects

AIMTo study effects of proanthocyanidins (PA) on contractile activity of isolated aortic smooth muscle in rats and rabbit platelet aggregation.

METHODSIsolated rat aortic muscle rings were adopted to observe the effects of PA on their contraction induced by noradrenaline (NA) or KCl, and their tensions were recorded by BL-310 experimental system of biological function. Rabbit platelet aggregation induced by arachidonic acid (AA), adenosine diphosphate (ADP), and collagen (Coll) was assayed by turbidimetry.

RESULTSPA could significantly inhibit the contraction induced by NA (10(-6) mol/L), low the concentration-response curves of NA and the maximal response on the endothelium-intact or endothelium-denuded aortic rings. But PA couldn't relax the aortic rings precontracted with KCl and had no influence on rabbit platelet aggregation induced by AA, ADP, and Coll.

CONCLUSIONPA can inhibit the contraction induced by NA but not by KCl on isolated rat aortic rings. It also has no influence on rabbit platelet aggregation.

Animals ; Aorta ; drug effects ; In Vitro Techniques ; Muscle Contraction ; drug effects ; Muscle, Smooth, Vascular ; drug effects ; physiology ; Norepinephrine ; pharmacology ; Platelet Aggregation ; drug effects ; Potassium Chloride ; pharmacology ; Proanthocyanidins ; pharmacology ; Rabbits ; Rats ; Rats, Wistar ; Seeds ; chemistry ; Vitis ; chemistry

Apoptosis ; drug effects ; Caspases ; physiology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; pathology ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; physiology ; Plant Extracts ; pharmacology ; Proanthocyanidins ; pharmacology ; Seeds ; chemistry ; Vitis ; chemistry

OBJECTIVETo investigate the inhibitory effect of grape seed extract (GSE) on the growth of prostate cancer PC-3 cells.

METHODSPC-3 cells were treated with GSE at the concentration of 100, 200 and 300 microg/ml for 24, 48 and 72 hours, respectively. The the inhibitory effect of GSE on the growth of the PC-3 cells and the kidney cells of SD rats was determined by MTT reduction assay, with primarily cultured kidney cells of 1-3 days old SD rats as the normal control.

RESULTSGSE significantly inhibited the growth of PC-3 cells in a concentration- and time-dependent manner, but had only a mild inhibitory effect on the kidney cells.

CONCLUSIONGSE inhibits the growth of prostate cancer PC-3 cells and can be used as a new drug for the treatment of prostate cancer.

Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Kidney ; cytology ; Male ; Plant Extracts ; pharmacology ; Prostatic Neoplasms ; pathology ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry ; Time Factors ; Vitis ; chemistry