Phytosterol content of sea buckthorn (Hippophae rhamnoides L.) seed oil: Extraction and identification

Phytosterols in sea buckthorn (Hippophae rhamnoides L.) seed oil extracted by cold pressing, hexane, and supercritical carbon dioxide were identified by GC–MS and FID. Compounds identified were campesterol, clerosterol, lanosterol, sitosterol, β-amyrin, sitostanol, Δ⁵-avenasterol, Δ²⁴⁽²⁸⁾-stigmasta-en-ol, α-amyrin, Δ^5,24(25)-stigmastadienol, lupeol, gramisterol, Δ⁷-sitosterol, cycloartenol, cycloeucalenol, Δ⁷-avenasterol, 28-methylobtusifoliol, 24-methylenecycloartanol, erythrodiol, citrostadienol, uvaol, and oleanol aldehyde. Sitosterol and Δ⁵-avenasterol were, quantitatively, the most important phytosterols. Total sterols and most individual sterols differed significantly (P ? 0.05) among all three extraction methods with supercritical carbon dioxide extracting the highest total sterol levels (1640 mg/100 g oil) and cold pressed the lowest levels (879 mg/100 g oil).     

Identification,quantification and antioxidant activity of acylated flavonol glycosides from sea buckthorn (Hippophae rhamnoides ssp. sinensis)

A novel acylated flavonol glycoside: isorhamnetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (1), together with two known acylated flavonol glycosides: quercetin (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (2) and kaempferol (3-O-[(6-O-E-sinapoyl)-β-d-glucopyranosyl-(1 → 2)]-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside) (3) were isolated from the n-butanol fraction of sea buckthorn (Hippophae rhamnoides ssp. sinensis) berries for the first time by chromatographic methods, and their structures were elucidated using UV, MS, ¹H and ¹³C NMR, and 2D NMR. Compounds 1–3 showed good scavenging activities, with respective IC₅₀ values of 8.91, 4.26 and 30.90 μM toward the 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical; respective Trolox equivalent antioxidant capacities of 2.89, 4.04 and 2.44 μM μM⁻¹ toward 2,2′-azino-bis-3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical. The quantitative analysis of the isolated acylated flavonol glycosides was performed by HPLC–DAD method. The contents of compounds 1–3 were in the range of 12.2–31.4, 4.0–25.3, 7.5–59.7 mg/100 g dried berries and 9.1–34.5, 75.1–182.1, 29.2–113.4 mg/100 g dried leaves, respectively.     

Inhibition of lipid oxidation and dynamics of polyphenol content in mechanically deboned meat supplemented with sea buckthorn (Hippophae rhamnoides) berry residues

Mechanically deboned meat (MDM) contains about 10 times more polyunsaturated fatty acids (PUFAs) and also more hemoproteins than hand deboned meat (HDM) and is essentially more susceptible to both chemical and biochemical oxidation. The oxidation, leading to the formation of potentially mutagenic and carcinogenic derivatives of PUFAs, can be inhibited by berry extracts rich in antioxidant polyphenols. Using the 2-thiobarbituric acid reactive substances (TBARS) method, we have established that the ethanol slurry of the juice-free solid residue of sea buckthorn (Hippophae rhamnoides – SB) berries inhibits oxidation of unsaturated fatty acids, of both chicken and turkey MDM. The polyphenols, mainly flavonols, responsible for this inhibition, are comparatively stable during short-term cooking and 6-day storage of cooked SB-MDMs at +6 °C. About half of the polyphenols are lost, obviously oxidised, during the storage of the uncooked samples of turkey 2%SB-MDM at +6 °C. The loss of polyphenols is much smaller in the case of chicken MDM, which is characterised by an in situ lower content of fatty acids, including the polyunsaturated ones. The liquid chromatography–diode array detection–tandem mass spectrometry (LC–DAD–ESI-MS/MS) method was used for identification and ranking of the potent polyphenolic antioxidants in the berry residue.     

Effects of drying method on the extraction yields and quality of oils from quebec sea buckthorn (Hippophaë rhamnoides L.) seeds and pulp

The effects of air-drying and freeze-drying on the extraction yields and quality of oils from Quebec sea buckthorn (cv. Indian-summer) seeds and pulp were studied. Oil extractions were carried out using hexane. Air-dried (ADS) and freeze-dried (FDS) seeds, gave a similar extraction yields (∼12% w/w), whereas those of air-dried (ADP) and freeze-dried (FDP) pulps were significantly different (35.9 ± 0.8 vs. 17.1 ± 0.6% w/w). Fatty acid analysis revealed that α-linolenic (37.2–39.6%), linoleic (32.4–34.2%) and oleic (13.1%) acids were the main fatty acids in seed oils, while pulp oils were rich in palmitoleic (39.9%), palmitic (35.4%) and linoleic (10.6%) acids. Lipid fractionation of crude oils, obtained by solid phase extraction (SPE), yielded mainly neutral lipids (93.9–95.8%). The peroxide values of seed and pulp oils were ca. 1.8 meq/kg and between 3.0 and 5.4 meq/kg, respectively. The melting behavior of seed and pulp oils showed multiple endothermic transitions, as observed normally in vegetable oils.     

Optimization of supercritical carbon dioxide extraction of sea buckthorn (Hippophaë thamnoides L.) oil using response surface methodology

Response surface methodology (RSM) was employed to optimize the conditions of supercritical CO₂ extraction of the whole berry oil from sea buckthorn. The effects of pressure, temperature, CO₂ flow rate and extraction time on the yield of oil, vitamin E and carotenoids were investigated. Results showed that the data were adequately fitted into three second-order polynomial models. The independent variables, the quadratics of pressure and extraction time, the interactions between pressure and temperature, pressure and extraction time, temperature and extraction time, as well as flow rate and extraction time had a significant effect on the yield of the oil, vitamin E or carotenoids. It was predicted that the optimum extraction conditions within the experimental ranges would be the extraction pressure of 27.6 MPa and temperature of 34.5 °C with flow rate of 17.0 L/h and extraction time of 82.0 min. Under such parameters, the yields of oil, vitamin E and carotenoids were predicted to be 208.0 g/kg, 288.7 mg/kg and 620.0 mg/kg dry sea buckthorn berry, respectively. Monounsaturated fatty acids were predominant in the whole berry oil, accounting for over 62% of the total fatty acids.     

Influence of origin,harvesting time and weather conditions on content of inositols and methylinositols in sea buckthorn (Hippophaë rhamnoides) berries

Inositols and methylinositols play an important role in human physiology. Inositols and methylinositols in berries of three subspecies of sea buckthorn (Hippophaë rhamnoides) were analysed using gas chromatography combined with a flame ionisation detector and mass spectrometry. The wild Chinese berries (H. rhamnoides ssp. sinensis) contained higher levels of l-quebrachitol (1l-2-O-methyl-chiro-inositol) and methyl-myo-inositol (average 615 and 58 mg/100 ml juice, respectively) than the Finnish (H. rhamnoides ssp. rhamnoides, 276 and 11 mg/100 ml juice, respectively) and the Russian (H. rhamnoides ssp. mongolica, 228 and 16 mg/100 ml juice, respectively) berries (P < 0.001). The content of myo-inositol was higher in the Chinese and the Russian berries than in the Finnish berries (26 and 20 mg/100 ml juice vs. 8 mg/100 ml juice, P < 0.001). In the Chinese berries, the contents of methyl-myo-inositol and l-quebrachitol increased, whereas that of myo-inositol decreased from late September to late November. The content of the l-quebrachitol in the Chinese berries correlated negatively with the air temperature and the number of frost-free days, suggesting a possible role of the compound in the cold resistance of sea buckthorn.     

Antimicrobial, antioxidant and phytochemical investigations of sea buckthorn (Hippophaë rhamnoides L.) leaf, stem, root and seed

The antimicrobial and antioxidant activities of crude ethanolic extract from Hippophaë rhamnoides L. (Elaeagnaceae) leaf, stem, root and seed, and their respective fractions, obtained by liquid-liquid extraction (LLE) using hexane (HF), ethyl acetate (EAF) and water (WF), were investigated. The crude extract was obtained by Pressurised Liquid Extraction (PLE), using ethanol at 100 bar and 60 °C. Antimicrobial activity was tested against food-borne and clinical microorganisms. Antioxidant activity was measured using the DPPH-radical scavenging and the ferric reducing antioxidant power (FRAP) assays. The phytochemical contents were examined by colorimetric methods. The results showed that crude extracts were active against Gram − and + strains, and that seed and root extracts were better radical scavengers than leaf and stem extracts. For all organs, the two activities tested were found to be higher in WF. These activities were correlated with the presence of phenolic compounds in active fractions. High Performance Thin Layer Chromatography (HPTLC) fingerprints confirmed presence of phenolic compounds in active extracts and fractions.     

Sugars, sugar alcohols, fruit acids, and ascorbic acid in wild Chinese sea buckthorn (Hippophaë rhamnoides ssp. sinensis) with special reference to influence of latitude and altitude

Wild berries of Hippophaë rhamnoides ssp. sinensis were collected from nine natural growth sites in China in three consecutive years in order to get an overall profile of the sugars, sugar alcohols, fruit acids, and ascorbic acid, and especially of the influence of the latitude and altitude of the growth place on these components. The contents of fructose, glucose, and l-quebrachitol in the berry juice varied in the ranges of 0.01-7.17, 0.05-7.85 and 0.21-1.09 g/100 mL, respectively, those of malic, quinic, and ascorbic acids were 1.55-8.84, 0.07-2.94, and 0.25-1.66 g/100 mL, respectively. The berries from Hebei and Inner Mongolia were characterized by high contents of sugars and l-quebrachitol and low contents of malic acid and ascorbic acid. In contrast, the berries from Sichuan and Qinghai contained lower contents of sugars and higher contents of malic acid and ascorbic acid than the berries from other growth areas. The berries from Sichuan differed considerably from others by the remarkably low contents of sugars and the exceptionally high contents of acids. The contents of fructose, glucose, and total sugar decreased as the altitude increased and as the latitude decreased (p < 0.05). In contrast, the contents of malic acid and ascorbic acid increased as the altitude increased and as the latitude decreased (p < 0.05). The contents of quinic acid and l-quebrachitol correlated strongly and positively with the latitude (p < 0.01).     

Evaluation of a simple and promising method for extraction of antioxidants from sea buckthorn (Hippophaë rhamnoides L.) berries: Pressurised solvent-free microwave assisted extraction

The pressurised solvent-free microwave assisted extraction (PSFME) technique has been developed and optimised for extraction of antioxidants from Hippophaë rhamnoides L. berries using a two-level full factorial design. The effects of factors (extraction time, irradiation power, number of cycles) and their first order interactions were evaluated from antioxidant activity of extracts using the 2,2′-diphenyl-1-picrylhydrazil (DPPH) free radical scavenging method, the ferric reducing ability of plasma (FRAP) assay, and the estimation of total phenolic content using the Folin–Ciocalteu method. The best extraction conditions were obtained, in a laboratory scale extractor of 50 mL filled with 4 g fresh berries, using a 1000 W microwave power applied during 50 s and repeated five cycles. PSFME was then compared to other common extraction techniques such as pressing, maceration and pressurised liquid extraction. It is appeared that PSFME leads to the most active and richest extract in phenolic content including molecules such as quercetin and isorhamnetin not extracted with other techniques. Furthermore PSFME respect green chemistry, it is rapid, cheap and does not need sample preparation and/or evaporation step.     

Inhibition effects and induction of apoptosis of flavonoids on the prostate cancer cell line PC-3 in vitro

Myricetin and myricitrin are naturally occurring flavonoids have been suggested to play a role in inhibition of proliferation and transformation of carcinogenic cell. However, the underlying molecular mechanisms of their activity have not yet to be revealed. The aim of the present study was to clarify the molecular mechanisms of apoptosis cell on the prostate cancer induced by myricetin, myricitrin, quercetin and quercitrin. The MTT assay confirmed that myricetin had the strongest inhibitory effect on human prostate cancer cell line PC-3, myricitrin was second, and quercitrin was the weakest. A noticeable synergistic effect was observed with the inhibition of cell proliferation when myricetin was used in combination with myricitrin. In the concentration range of 37.5–300 μmol/L, the inhibitory effects of these flavonoids were enhanced with increasing dose and treatment time. The acridine orange analysis and annexin V–FITC/PI double-staining results confirmed that myricetin and myricitrin were effective in inducing PC-3 cell apoptosis. The results showed that myricetin was more effective than myricitrin in inducing cell apoptosis. The apoptosis rate increased with increasing flavonoid concentration in a dose dependent manner. A synergistic effect was observed on the apoptosis rate when myricetin was used in combination with myricitrin.     

Variability and the genotypic effect on antioxidant activity,total phenolics,carotenoids and ascorbic acid content in seventeen natural population of Seabuckthorn (Hippophae rhamnoides L.) from trans-Himalaya

Seventeen natural population of Seabuckthorn (SBT), which comprised 187 plants from trans-Himalaya, were studied to find out variability and genotypic effect on total phenolic content (TPC), total antioxidant capacity (TAC), ascorbic acid and carotenoids content in fruit pulp. The fruits were found to be rich in TPC ranging from 964 to 10,704 mg gallic acid equivalent/100 g. The free radical-scavenging activity in terms of inhibitory concentration (IC₅₀) ranged from 0.7 to 9.1 mg/ml and ferric reducing antioxidant potential (FRAP) from 180 to 1355 FeSO₄·7H₂O μg/ml. The ascorbic acid and carotenoids content ranged from 56 to 3909 mg/100 g and 0.1–14.4 mg/100 g, respectively. A variation of 1–11 fold in TPC, 1–14 folds in IC₅₀ by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and 1–8 fold in ferric reducing antioxidant potential, 1–70 fold in ascorbic acid content and 1–206 fold in carotenoid content among the examined fruit across 17 populations underlines the important role played by genetic background and the geographical location for determining the health promoting compounds. Significant correlation was observed between TPC, IC₅₀, FRAP, carotenoids, ascorbic acid, fruit lightness (L*) and plant height. Among the 20 morphological traits studied, fruit colour and plant height showed positive correlation with the health promoting compounds.     

Araçá (Psidium cattleianum Sabine) fruit extracts with antioxidant and antimicrobial activities and antiproliferative effect on human cancer cells

Araçá or strawberry guava (Psidium cattleianum Sabine) is an attractive tasty small fruit native to temperate zones of Brazil. In this study, functional chemical constituents and the nutraceutical and therapeutic potential of aqueous and acetone extracts of red and yellow accessions of araçá were characterised. While carotenes, ascorbic acid, and anthocyanins were present as minor constituents, araçá fruit presented high levels of phenolic compounds (up to 768 mg 100 g⁻¹ fresh fruit pulp, ffp), particularly (−)-epicatechin (up to 2.7 mg g⁻¹ ffp), which were in general more efficiently extracted with acetone. Abundance of phenolic compounds was positively correlated with antioxidant activity, antimicrobial and antiproliferative effects.     

Quantification of gallic acid and ellagic acid from longan (Dimocarpus longan Lour.) seed and mango (Mangifera indica L.) kernel and their effects on antioxidant activity

Gallic acid (GA) and ellagic acid (EA) have been identified in longan seed and mango kernel by the use of reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with photodiode array detection (DAD). The ethanolic extract of longan seed contained 23.3 and 156 mg/100 seeds of GA and EA, respectively. The ethanolic extracts of mango kernel contained approximately 87% more GA than the longan seed ethanolic extracts but about 32% less EA. After heat treatment and acid hydrolysis, mango kernel had higher concentrations of GA and EA, contributing to more potent antioxidant activity. The results demonstrated rich sources of GA and EA in longan seed and mango kernel which might provide a novel source of these natural antioxidants.     

Induction of the cell cycle arrest and apoptosis by flavonoids isolated from Korean Citrus aurantium L. in non-small-cell lung cancer cells

This study investigated the anti-proliferative and apoptotic effect of flavonoids isolated from Korean Citrus aurantium L. using A549 lung cancer cells. Flavonoids potently inhibited of A549 cells in a dose-dependent manner, whereas flavonoids had a weak inhibitory effect on proliferation of WI-38 cells. Flow cytometry and Western blot analysis showed that flavonoids induced cell cycle arrest at the G2/M checkpoint by controlling the proteins expression level of cyclin B1, cdc2, cdc25c and p21^WAF1/CIP1. Also, flavonoids induced apoptosis through the regulation of the expression of caspases, cleaved PARP and Bax/Bcl-xL ratio. The activity of caspase-3 on A549 cells increased in a dose-dependent manner. These results clearly indicated that the anti-cancer effect of flavonoids on A549 cells follows multiple cellular pathways through G2/M arrest and the induction of apoptosis.     

Phenolic acid contents of kale (Brassica oleraceae L. var. acephala DC.) extracts and their antioxidant and antibacterial activities

Nine phenolic acids were identified and quantified by HPLC–MS in leaves and 10 in seeds of kale (black cabbage). The free, ester (methanol-soluble), glycoside and ester-bound (methanol-insoluble) phenolic acid contents of the leaves were 487, 532, 4989 and 6402 ng/g fresh weight, respectively. Ferulic and caffeic acids (total contents; 4269 and 4887 ng/g, respectively) were the most abundant. The seed contents of these fractions were 1993, 1477, 1231 and 4909 ng/g dry weight (DW), respectively, and sinapic acid was the most abundant (5037 ng/g DW). The fractions’ total phenolic contents, determined colorimetrically, were highly correlated with their DPPH scavenging capacity, and in antimicrobial activity assays, with nine test organisms representing a wide array of taxa, all of the fractions were effective against Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis and (most strongly) Moraxella catarrhalis. Antimicrobial and antioxidant activities of kale phenolics in free and conjugated forms are discussed.     

Effect of candying on cell wall polysaccharides of plums (Prunus domestica L.) and influence of cell wall enzymes

“Ameixa d’Elvas” is a candied plum (Prunus domestica L.) produced by a traditional process, using fruits of a specific ‘greengage’ variety, “Rainha Cláudia Verde”. The candying process consists of boiling the intact plums in water for 15 min and then putting them in sugar syrup, which is successively concentrated until 75 °Brix. Although a loss of intercellular adhesion of parenchyma cells after boiling is observed, candied plums are able to recover their cell-to-cell adhesion, giving a final tissue with a consistency similar to that observed for the fresh fruit. In order to explain this observation, cell wall polysaccharides of plums harvested in two orchards, Vila Viçosa (VV) and Cano (CA), from the same geographic region and at the same stage of ripening, were analysed fresh, boiled and candied. Plum cell walls are composed mainly of pectic polysaccharides and cellulose that, during the boiling step, are degraded and solubilised. Highly esterified pectic polysaccharides undergo gelation inside the fruits in the presence of sucrose, leading to the recovery of the fruit’s consistency. During the candying process diffusion of these methylesterified pectic polysaccharides to the sucrose syrup increase the syrup viscosity. The activity of pectin methylesterase, polygalacturonase, and cellulase of fresh fruits explains the observed higher extension of degradation of cell wall polysaccharides of the CA plum tissues after boiling. This higher degradation seems to prevent the complete recovery of the parenchyma cell structure, which was observed for the less degraded polysaccharides of VV plums.