不同花色苷代表性成分的分离鉴定及体外抗氧化活性

本实验以紫甘薯、黑枸杞、黑加仑和桑葚花色苷提取物为原料,制备其单体花色苷组分并研究其体外抗氧化性质。选取每种花色苷中含量较高,分子量居中,具有代表性的单体化合物作为目标组分,采用高速逆流色谱制备分离四种来源的花色苷。选用甲基叔丁基醚-正丁醇-乙腈-水-三氟乙酸作为溶剂体系,流速设定为5 mL/min,转速为850 r/min,分离得到高纯度花色苷单体化合物。采用分光光度法、HPLC-MS法分析测定花色苷含量及主要组成,用DPPH自由基、羟自由基清除能力和总还原力的测定分析其体外抗氧化活性。结果表明,四种来源的花色苷中代表性的成分依次为芍药素-3-咖啡酰-阿魏酰槐糖苷-5-葡萄糖苷、矮牵牛素-3-O-对香豆酰芸香糖苷-5-O-葡萄糖苷、矢车菊-3-芸香糖苷和矢车菊-3-O-葡萄糖苷,它们均具有良好的体外抗氧化活性。     

LC‐MS identification of anthocyanins in boysenberry extract and anthocyanin metabolites in human urine following dosing

The anthocyanin composition of boysenberry (Rubusloganbaccus × baileyanus Britt) extract was determined by LC‐ESI‐MS. Four anthocyanins were identified, all comprising a cyanidin‐anthocyanidin‐type skeleton. The two major components were identified as the disaccharide cyanidin‐3‐O‐sophoroside and the monosaccharide cyanidin‐3‐O‐glucoside. The two less abundant components were identified as the rutinosides, cyanidin‐3‐O‐2^G‐glucosylrutinoside and cyanidin‐3‐O‐rutinoside, respectively. These same four anthocyanins were also detected in human urine following a dosing study with boysenberry extract indicating that glycosylated anthocyanins can be absorbed from the gut and excreted intact in the urine. Several anthocyanin metabolites were also detected in the urine and were identified by LC‐ESI‐MS as monoglucuronides of peonidin, cyanidin and pelargonidin. The results suggest that anthocyanins consumed as part of a diet are bioavailable and are present as intact or metabolized forms in the body. Copyright © 2004 Society of Chemical Industry     

Characterisation of phenols and antioxidant and hypolipidaemic activities of Lethariella cladonioides

BACKGROUND: This study aimed to investigate the antioxidant and hypolipidaemic activities of an ethanol extract of Lethariella cladonioides (Nyl.) Krog (EE) and to characterise its chemical constituents. RESULTS: Nine phenols were identified as canarione, thamnolic acid, squamatic acid, vermicularin, norstictic acid, baeomycesis acid, lecanoric acid, barbatinic acid and usnic acid from analysis of EE by using high‐performance liquid chromatography with a diode array detector‐mass spectrometry. In antioxidant analysis in vitro, the highest scavenging rate of EEs on the 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radicals, superoxide anion, hydroxyl radicals and hydrogen peroxide was 81.55 ± 1.95%, 81.84 ± 4.00%, 74.28 ± 3.71% and 74.28 ± 3.71%, respectively. Meanwhile, after administration of EE for 6 weeks in high fat/cholesterol diet mice, the most significant reduction in levels of serum triglyceride, serum total cholesterol, serum low density lipoprotein cholesterol and liver malondialdehyde were 24%, 20%, 15% and 35%, respectively. The most significant increase in levels of serum high density lipoprotein cholesterol and liver superoxide dismutase was 35% and 88%, respectively. CONCLUSION: L. cladonioides possesses strong antioxidant and hypolipidaemic activities. Copyright © 2011 Society of Chemical Industry     

Optimization of extraction conditions of total phenolics, antioxidant activities, and anthocyanin of oregano, thyme, terebinth, and pomegranate

The purpose of this study was to evaluate the total phenolic extracts and antioxidant activity and anthocyanins of varieties of the investigated plants. These plants include oregano, thyme, terebinth, and pomegranate. The optimum extraction conditions including temperature and solvent of the extraction process itself were investigated. Total phenolic and anthocyanin extracts were examined according to Folin-Ciocalteu assay and Rabino and Mancinelli method, respectively. The effect of different extracting solvents and temperatures on extracts of phenolic compounds and anthocyanins were studied. Plant samples were evaluated for their antioxidant chemical activity by 2, 2-diphenyl-1-picrylhydrazl assay, to determine their potential as a source of natural antioxidant. Results showed that all tested plants exhibited appreciable amounts of phenolic compounds. The methanolic extract (60 °C) of sour pomegranate peel contained the highest phenolic extract (4952.4 mg/100 g of dry weight). Terebinth green seed had the lowest phenolic extract (599.4 mg/100 g of dry weight). Anthocyanins ranged between 3.5 (terebinth red seed) and 0.2 mg/100 g of dry material (thyme). Significant effect of different extracting solvents and temperatures on total phenolics and anthocyanin extracts were found. The methanol and 60 °C of extraction conditions found to be the best for extracting phenolic compounds. The distilled water and 60 °C extraction conditions found to be the best for extracting anthocyanin.     

Storage effects on anthocyanins, phenolics and antioxidant activity of thermally processed conventional and organic blueberries

BACKGROUND: Consumer demand for products rich in phytochemicals is increasing as a result of greater awareness of their potential health benefits. However, processed products are stored for long‐term and the phytochemicals are susceptible to degradation during storage. The objective of this study was to assess the storage effects on phytochemicals in thermally processed blueberries. Thermally processed canned berries and juice/puree were analysed for phytochemicals during their long‐term storage. RESULTS: The phytochemical retention of thermally processed blueberries during storage was not influenced by production system (conventional versus organic). During 13 months of storage, total anthocyanins, total phenolics and total antioxidant activity in canned blueberry solids decreased by up to 86, 69 and 52% respectively. In canned blueberry syrup, total anthocyanins and total antioxidant activity decreased by up to 68 and 15% respectively, while total phenolic content increased by up to 117%. Similar trends in phytochemical content were observed in juice/puree stored for 4 months. The extent of changes in phytochemicals of thermally processed blueberries during storage was significantly influenced by blanching. CONCLUSION: Long‐term storage of thermally processed blueberries had varying degrees of influence on degradation of total anthocyanins, total phenolics and total antioxidant activity. Blanching before thermal processing helped to preserve the phytochemicals during storage of blueberries. Copyright © 2011 Society of Chemical Industry     

Effects of gastrointestinal digestion models in vitro on phenolic compounds and antioxidant activity of juçara (Euterpe edulis)

This study evaluated the effects of different gastrointestinal digestion models in vitro on the bioaccessibility of phenolics and antioxidant activity in juçara frozen pulp. In the sequence, method 3 was applied to juçara fruit in three different stages of maturation (vitrin – reddish fruits, mature – purple fruits, tuíra – deep purple fruits). In the method applied, the final pH adopted was 5.0, in order to avoid interference in the assay used to determine the antioxidant activity, and BHT was used to prevent excessive oxidation in the system. In this method, higher values for antioxidant activity were obtained (3574.95–3719.10 μmol L^?1 Trolox 100 g^?1 pulp) compared with the other two methods tested (1969.14–3034.74 μmol L^?1 Trolox 100 g^?1 pulp). In relation to juçara fruit, the mature stage was found to be ideal for processing, showing generally higher values of the bioaccessibility for phenolics and antioxidant activity compared to other maturation stages.     

Stability and antioxidant activity of anthocyanins from purple sweet potato (Ipomoea batatas L. cultivar Eshu No. 8) subjected to simulated in vitro gastrointestinal digestion

The study was aimed to investigate the stability and antioxidant activities of anthocyanins obtained from purple sweet potato via an in vitro digestion system. Three fractions of anthocyanin-rich extracts were obtained via ODS packing column. Anthocyanins and copigment (primary phenolic acids) of fraction 2 were investigated during simulated an in vitro gastrointestinal digestion. Anthocyanins and copigment were recognised by UPLC-LTQ-MS/MS. Besides, anthocyanins were found effectively stable under the acidic gastric digestion conditions. However, the anthocyanins recovery was greatly decreased at around 10% after intestinal digestion. An association between the type, number of acylated group and stability to intestinal digestion was found. Di-acylated anthocyanins possessed higher stability compared with mono-acylated anthocyanins and the stability of acylated group in digestion process followed the order of p-hydroxybenzoyl > feruloyl > caffeoyl. However, there was no much difference in copigment content which was found during digestion process. Moreover, the radical scavenging ability and xanthine oxidase (XO) inhibition showed that the digestion products possessed good biological activities mainly due to its anthocyanin composition.     

‘蓓蕾’蓝靛果中花色苷组成鉴定及抗氧化能力比较分析

对蓝靛果中花色苷的组成进行鉴定,并对其抗氧化能力进行比较分析。实验以蓝靛果（‘蓓蕾’品种）为原料,采用有机溶剂60%乙醇（0.1%盐酸酸化）溶液,超声辅助提取90 min;利用D101大孔树脂对获得的粗提物进行纯化,之后冷冻干燥制得粉末物质。通过pH示差法和福林-酚法分别测定总花色苷含量和总多酚含量,分别为（353.35±0.79）、（474.01±2.12）mg/g;并用高效液相色谱-质谱联用法对花色苷组成进行鉴定,共发现11 种花色苷,其中矢车菊-3-葡萄糖苷为主要花色苷（90.679%）。此外,实验还通过总抗氧化能力测定和2,2’-联氨-二（3-乙基苯并噻唑-6-磺酸）二铵盐自由基、1,1-二苯基-2-三硝基苯肼自由基清除能力测定,比较分析蓝靛果花色苷提取物、矢车菊-3-葡萄糖苷、VC的抗氧化能力,结果表明,3 种物质的抗氧化能力排序为：矢车菊-3-葡萄糖苷＞花色苷提取物＞VC。     

Oregano: chemical analysis and evaluation of its antimalarial, antioxidant, and cytotoxic activities

Abstract: GC‐FID and GC‐MS analysis of essential oil from oregano leaves (Origanum compactum) resulted in the identification of 46 compounds, representing more than 98% of the total composition. Carvacrol was the predominant compound (36.46%), followed by thymol (29.74%) and p‐cymene (24.31%). Serial extractions with petroleum ether, ethyl acetate, ethanol, and water were performed on aerials parts of Origanum compactum. In these extracts, different chemical families were characterized: polyphenols (gallic acid equivalent 21.2 to 858.3 g/kg), tannins (catechin equivalent 12.4 to 510.3 g/kg), anthocyanins (cyanidin equivalent 0.38 to 5.63 mg/kg), and flavonoids (quercetin equivalent 14.5 to 54.7 g/kg). The samples (essential oil and extracts) were subjected to a screening for antioxidant (DPPH and ABTS assays) and antimalarial activities and against human breast cancer cells. The essential oil showed a higher antioxidant activity with an IC₅₀= 2 ± 0.1 mg/L. Among the extracts, the aqueous extract had the highest antioxidant activity with an IC₅₀= 4.8 ± 0.2 mg/L (DPPH assay). Concerning antimalarial activity, Origanum compactum essential oil and ethyl acetate extract showed the best results with an IC₅₀ of 34 and 33 mg/mL, respectively. In addition, ethyl acetate extract (30 mg/L) and ethanol extract (56 mg/L) showed activity against human breast cancer cells (MCF7). The oregano essential oil was considered to be nontoxic.     

Extraction,isolation, characterisation,antioxidant and anti-fatigue activities of Pleurotus eryngii polysaccharides

Pleurotus eryngii polysaccharide (PEP) was obtained using hot water extraction and ethanol precipitation. Three purified polysaccharide fractions (namely PEP1-A, PEP2-A and PEP3-A) were obtained from PEP using DEAE-cellulose-52 chromatography and a gel permeation Sephadex G-100 column. Firstly, this paper examined the characterisation of PEP1-A, PEP2-A and PEP3-A. The corresponding molecular weights were 5.378 × 10⁵, 9.506 × 10⁶ and 4.975 × 10⁵ Da, respectively. PEP1-A, PEP2-A and PEP3-A had similar monosaccharide compositions. PEP1-A was β-configuration, and PEP2-A and PEP3-A were α-configuration. PEP1-A, PEP2-A and PEP3-A had pyran-type rings, (1 → 3) glucose and (1 → 6) galactose linkages. Secondly, PEP, PEP1-A, PEP2-A and PEP3-A possessed antioxidant activities, and PEP was best. Therefore, only PEP was used to study its anti-fatigue activity in vivo. The result proved that PEP had anti-fatigue activity. PEP could be used as a valuable natural food supplement for preventing anti-fatigue or functional food.     

GC/MS analysis and antimicrobial and antioxidant activities of essential oil of Eucalyptus radiata

BACKGROUND: The essential oil from Eucalyptus radiata leaves collected in Tunisia was extracted by steam distillation and analysed by gas chromatography/flame ionisation detection and gas chromatography/mass spectrometry. Its antioxidant and antiradical properties were evaluated by the 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) assays. The antimicrobial activity of the oil was examined in vitro against two plant‐pathogenic bacteria (four strains each of Agrobacterium tumefaciens and Pseudomonas savastanoi pv. savastanoi) and two plant‐pathogenic fungi (Fusarium solani and Rhizoctonia solani). RESULTS: Thirty‐five compounds were identified and quantified in the essential oil, the major ones being 1,8‐cineole (69.53%), α‐pinene (11.94%) and trans‐pinocarveol (4.81%). Medium antioxidant activity was found in the ABTS assay (IC₅₀ = 484.3 ± 17.3 mg L^?1), whereas no significant free radical‐scavenging activity was detected in the DPPH assay (IC₅₀ > 10 000 mg L^?1). The antimicrobial assays showed that the oil exhibited a high level of activity against A. tumefaciens and R. solani, with minimum inhibitory concentrations ranging between 750 and 1000 µL L^?1. However, it was less efficient against P. savastanoi pv. savastanoi and F. solani. CONCLUSION: The results indicate that the essential oil of E. radiata, with a high content of terpenic compounds, exhibits significant antimicrobial activity against strains of A. tumefaciens and the fungus R. solani and may therefore be useful for their control. Copyright © 2009 Society of Chemical Industry     

灰树花抗氧化活性多酚的提取纯化及其鉴定

以抗氧化活性为指标,通过单因素试验和正交试验对灰树花多酚提取工艺进行优化,确定最佳提取条件为提取时间2.5 h,提取温度70 ℃,料液比1∶50（g∶mL）。提取的多酚经大孔树脂NKA-Ⅱ动态吸附和解吸得到3个不同多酚组分,分别为体积分数为20%、40%及60%乙醇洗脱物。以DPPH自由基清除率、羟基自由基清除率以及总还原力为指标测定不同组分的抗氧化活性,结果表明,体积分数为40%乙醇洗脱物的抗氧化作用最强。采用高效液相色谱-串联质谱对体积分数为40%乙醇洗脱物多酚组分进行分离鉴定,通过对应的质谱结果分析可推测其中的多酚组分包括2-羟基丁二酸、香豆酸、3-羟基白藜芦醇、白藜芦醇、二羟基苯乙酸、咖啡酸和4-羟基苯甲酸等物质。     

Phenolic contents and antioxidant activities of bitter gourd (Momordica charantia L.) leaf, stem and fruit fraction extracts in vitro

Bitter gourd (Momordica charantia L.) has long been regarded as a food and medicinal plant. We investigated the antioxidant activity of the water extract of leaf, stem and fruit fractions by several in vitro systems of assay, namely DPPH radical-scavenging activity, hydroxyl radical-scavenging activity, β-carotene–linoleate bleaching assay, ferric reducing/antioxidant power (FRAP) assay and total antioxidant capacity. Total phenolic content was measured by Folin–Ciocalteu reagent. Identification of phenolic compounds was achieved using HPLC with the UV-diode array detection. The extracts of different fractions were found to have different levels of antioxidant activity in the systems tested. The leaf extract showed the highest value of antioxidant activity, based on DPPH radical-scavenging activity and ferric reducing power, while the green fruit extract showed the highest value of antioxidant activity, based on hydroxyl radical-scavenging activity, β-carotene–linoleate bleaching assay and total antioxidant capacity. The predominant phenolic compounds were gallic acid, followed by caffeic acid and catechin. The present study demonstrated that the water extract fractions of bitter gourd have different responses with different antioxidant methods. Total phenol content was shown to provide the highest association with FRAP assay in this present study (R² = 0.948).     

Flavonoid glycosides and antioxidant capacity of various blackberry,blueberry and red grape genotypes determined by high‐performance liquid chromatography/mass spectrometry

Anthocyanin and flavonol glycosides of various blackberry, blueberry and red wine grape genotypes were identified and measured by a high‐performance liquid chromatographic (HPLC) separation method with photodiode array (PDA) and mass spectrometric (MS) detection. With this method, two distinct elution regions of anthocyanins and flavonols were obtained with near baseline separation of most compounds. Blackberry, blueberry and red wine grape genotypes varied markedly in total anthocyanins and total flavonols as well as oxygen radical absorbance capacity (ORAC). The respective ranges of total anthocyanin (TA) and total flavonol (TF) contents of tested samples were: blackberries, 1143.9–2415.4 and 102.0–160.2 mg kg^?1; blueberries, 1435.2–8227.3 and 172.5–327.5 mg kg^?1; and red wine grapes, 380.9–7904.7 and 21.0–322.2 mg kg^?1. Antioxidant activities and contents of total anthocyanins and total flavonols in blackberries, blueberries and red wine grapes were highly correlated, with linear relationships between ORAC and TA (r_xy = 0.94) and TF (r_xy = 0.90) for grapes, TA (r_xy = 0.95) for blueberries and TA (r_xy = 0.74) for blackberries. Copyright © 2004 Society of Chemical Industry     

Ultrasound-assisted enzymatic extraction of anthocyanins from grape skins: optimization,identification, and antitumor activity

Grape skins produced during the grape juice production and processing contain abundant anthocyanins and other active compounds. Consequently, this study optimized the extraction conditions for ultrasound-assisted enzymatic extraction (UAEE) of anthocyanins from grape skins via response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest anthocyanins yield (3.01 ± 0.04) mg/g from grape skins by UAEE were obtained under an extraction temperature of 50 °C, ultrasonic power of 400 W, pectinase dosage of 0.16%, and extraction time of 28 min. The AB-8 macroporous resin combined Sephadex LH-20 techniques were further employed to purify the anthocyanins extracts obtained under optimum extraction conditions (AEOEC), and the main anthocyanins were identified using high-performance liquid chromatography tandem mass spectrometry. The purified anthocyanins contained two anthocyanins in terms of delphinidin-3,5-O-diglucoside and cyanidin-3-O-rutinoside with purity of 91.35% and 92.64%, respectively. Ultimately, we further evaluated the antitumor activity of AEOEC and two purified anthocyanins on breast cancer. The results indicated that the antitumor effect of AEOEC on breast cancer MCF-7 cells was better than that of two purified anthocyanins. In addition, AEOEC could memorably increase intracellular reactive oxygen species levels and apoptosis of MCF-7 cells, and arrest MCF-7 cells in the G2/M phases. The findings provide an effective and feasible method for anthocyanins extraction and reduce the environmental burden of this waste.     

葵花籽壳黑色素提取鉴定及抗氧化性研究

利用碱提酸沉和超声波等方法从葵花籽壳中提取黑色素,Folin-Ciocalteu法测定葵花籽壳黑色素粗提取物中总酚的含量,采用单因素和正交实验确定葵花籽壳黑色素提取的最佳工艺条件,紫外和红外光谱法初步鉴定黑色素;以VC、没食子酸为对照,测定黑色素粗提物与纯化黑色素的还原力和清除DPPH.的能力。结果表明,碱提酸沉葵花籽壳黑色素粗提物呈黑色、无光泽,最优提取条件下氢氧化钠浓度为0.5mol/L,提取时间1.5h,料液比1:25,提取温度25℃下,黑色素粗提物的得率为12.37%(RSD2.14%);纯化的黑色素呈黑色、有金属光泽,得率为2.07%(RSD9.27%);粗提物总酚含量以没食子酸计为0.48%。葵花籽壳黑色素亚硫酸钠溶液在292nm波长有最大吸收峰,且吸光度与黑色素的浓度呈正相关,红外光谱图有黑色素红外吸收特征。黑色素的还原力和清除DPPH.自由基的能力明显比VC低,在同一浓度下,纯化前后黑色素的抗氧化性能力无明显改变。