黄原胶对体外模拟消化过程中4 种竹叶黄酮生物可及性的影响

目的：竹叶碳苷黄酮在胃肠道消化过程中易于降解,导致其生物可及性低,因此,本研究拟探究黄原胶存在时,4 种碳苷黄酮（异荭草苷、荭草苷、异牡荆素、牡荆素）在各模拟消化阶段生物可及性的变化情况。方法：利用体外模拟消化模型结合超高效液相色谱技术,分析在有/无黄原胶存在的条件下,竹叶提取物中4 种碳苷黄酮质量浓度的变化情况。结果：相较于直接消化样品,添加黄原胶的样品可以明显减弱模拟消化过程中碳苷黄酮的降解;口腔、胃、肠道模拟消化对竹叶碳苷黄酮的生物可及性影响存在差异,黄原胶的加入可以明显提高4 种碳苷黄酮,尤其是牡荆素和异牡荆素的生物可及性,模拟肠道消化后,黄原胶保护组的异牡荆素和牡荆素质量浓度分别降低了62%和59%,远低于直接消化组（分别降低84%和80%）。结论：黄原胶可以明显提升体外模拟消化过程中竹叶碳苷黄酮的生物可及性,对于相关功能产品的开发具有良好的应用价值。     

Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin

Orientin, vitexin and other flavone C-glycosides are functional ingredients abundant in trollflowers. In this study, an efficient method for enrichment of these ingredients from the flowers of Trollius chinensis Bunge was developed using macroporous resin. Separation characteristics of six typical macroporous resins were investigated by static adsorption/desorption and dynamic separation experiments, and HPD450 was selected as optimal one. Dynamic adsorption/desorption experiments on HPD450 columns were conducted to obtain the optimal parameters, followed by a scale-up experiment. Six fractions, TC-1–TC-3 together with their acid hydrolysates were further prepared to evaluate their antioxidant capacities by 2,2-diphenyl-1-picrylhydrazyl radical and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate) radical cation scavenging assays. They all have notable concentration-dependent antioxidant activities, with TC-1 showing strong antioxidant capacity higher than orientin. The separation process was high-efficient, low-cost and environmental-friendly, which could afford a potential approach for industrial applications.     

Variation in contents of phenolic compounds during growth and post-harvest storage of pigeon pea seedlings

The variation in contents of seven phenolic compounds in pigeon pea seedlings during growth and storage was investigated. Maximum contents of vitexin, isovitexin and orientin were found in leaves growing 40 days, which were 0.99 ± 0.06, 6.63 ± 0.35 and 30.89 ± 1.92 mg/g DW. Apigenin and luteolin were extensively distributed in leaves, stems and roots. Pinostrobin and cajaninstilbene acid were mainly accumulated in leaves, the peak values 3.53 ± 0.18 and 2.49 ± 0.13 mg/g DW appeared at the 60th day. Slight and steady increases of seven phenolic compounds were found in room temperature (25 °C) stored pigeon pea leaves up to 120 days. The highest accumulation of seven phenolic compounds at chilling temperature (4 °C) was observed at the 45th day, after which the contents decreased sharply. The stems extracts exhibited more efficient DPPH radical-scavenging ability while the roots extracts demonstrated the strongest lipid peroxidation inhibitory activity.     

Antioxidant Properties of Flavone C-Glycosides from Atractylodes japonica Leaves in Human Low-density Lipoprotein Oxidation

ABSTRACT: Three antioxidant flavone C -glucosides, isoorientin, vitexin, and isovitexin, were identified for the 1st time from Atractylodes japonica leaves by an activity-guided fractionation with various analytical techniques including column chromatography, high-performance liquid chromatography (HPLC), fast atom bombardment mass spectrometry (FAB-MS), and nuclear magnetic resonance (NMR). Hydroxyl and superoxide anion radicals scavenging activities of these compounds were examined using electron spin resonance (ESR). Inhibitory activities of these compounds on human low-density lipoprotein (LDL) oxidation were evaluated by malondialdehyde (MDA) and thiobarbituric acid-reactive substances (TBARS), both representing intermediates of lipid peroxidation. These flavone C -glucosides displayed about 50% of scavenging activity against hydroxyl radicals at the concentrations below 10 μ M . The superoxide dismutase (SOD)-equivalent activities of isoorientin, vitexin, and isovitexin at 1 m M were 31.37, 2.71, and 2.63 unit/mL, respectively. Isoorientin at 1 μ M exhibited over 60% inhibition of MDA formed during copper-mediated human LDL oxidation. Amounts of free MDA in LDL treated with isoorientin, vitexin, isovitexin, and control were 20.06, 40.73, 34.08, and 48.03 n M /mg protein, respectively. These compounds also prolonged the lag phase time of the conjugated diene formation. There was a positive correlation between the free radical scavenging activities and the inhibitory effects on the LDL oxidation of these compounds. These results suggest that the flavone C -glucosides isolated from the leaves of A japonica possess beneficial antioxidant properties against free radicals as well as LDL oxidation.     

Antioxidants of therapeutic relevance in COPD from the neotropical blueberry Anthopterus wardii

Four flavone C-glycosides, isoorientin (1), orientin (2), vitexin (3), and isovitexin (4), were isolated from the neotropical blueberry of Anthopterus wardii, a so-called “superfruit”, using antioxidant activity-guided fractionation. A dose-response relationship of compounds 1-4 was determined for their anti-inflammatory activity against interleukin-8 (IL-8) and for the inhibition of matrix metalloproteinase-1 (MMP-1) expression, an inflammatory marker for chronic obstructive pulmonary disease. The four flavone C-glycosides exhibited inhibitory activity against IL-8 production and MMP-1 expression, with compounds 1, 3, and 4 having the most potent inhibitory activities in both assays at 100 μg/ml. The structures of compounds 1-4 were determined by spectroscopic methods. These flavone C-glycosides are reported for the first time in the Anthopterus genus.     

Inhibitory effect of mung bean extract and its constituents vitexin and isovitexin on the formation of advanced glycation endproducts

The anti-glycation activity of four kinds of beans including mung bean, black bean, soybean and cowpea were evaluated. Aqueous alcohol extract of mung bean exhibited the strongest inhibitory activity against the formation of fluorescent advanced glycation endproducts (AGEs) in a bovine serum albumin (BSA)-glucose model, and the inhibitory activities of extracts of the four beans were found to be highly correlated with their total phenolic contents (R² = 0.95). Subsequent HPLC analysis of mung bean extract revealed two major phenolics which were purified and identified as vitexin and isovitexin by spectral methods. In the anti-glycation assays, both vitexin and isovitexin showed significant inhibitory activities against the formation of AGEs induced by glucose or methylglyoxal with efficacies of over 85% at 100 μM. In another assay, vitexin and isovitexin failed to directly trap reactive carbonyl species, such as methylglyoxal, suggesting that their anti-glycation activities may mainly be due to their free radical scavenging capacity.     

Phytochemical composition and thermal stability of two commercial açai species, Euterpe oleracea and Euterpe precatoria

Açai fruit are native to the Amazon region of South America and two predominant species are commercially exported as fruit pulps for use in food and beverage applications. Detailed characterisation of the polyphenolic compounds present in the de-seeded fruits of Euterpe oleracea and Euterpe precatoria species were conducted by HPLC–ESI–MSⁿ analyses and their thermal stability and overall influence on antioxidant capacity were determined. Anthocyanins were the predominant polyphenolics in both E. oleracea (2247 ± 23 mg/kg) and E. precatoria (3,458 ± 16 mg/kg) species, and accounted for nearly 90% of the trolox equivalent antioxidant capacity in both E. oleracea (87.4 ± 4.4 μmol TE/g) and E. precatoria (114 ± 6.9 μmol TE/g) fruits. Various flavones, including homoorientin, orientin, taxifolin deoxyhexose and isovitexin; various flavanol derivatives, including (+)-catechin, (−)-epicatechin, procyanidin dimers and trimers, and phenolic acids, including protocatechuic, p-hydroxybenzoic, vanillic, syringic and ferulic acids, were also present in both species. Thermal stability of these compounds was evaluated, following a thermal holding cycle (80 °C for up to 60 min) in the presence and absence of oxygen. Both species experienced only minor changes (<5%) in non-anthocyanin polyphenolic contents during all thermal processes whereas 34 ± 2.3% of anthocyanins in E. oleracea and 10.3 ± 1.1% of anthocyanins in E. precatoria were lost under these conditions, regardless of the presence of oxygen. Proportional decreases (10–25%) in antioxidant capacity accompanied the anthocyanin changes. Results suggest that both açai species are characterised by similar polyphenolic profiles, comparable antioxidant capacities, yet only moderate phytochemical stability during heating.     

高效液相色谱法测定粽子中的黄酮类化合物及其迁移规律

建立天然包装材料箬叶及粽米中荭草苷、异荭草苷、牡荆苷、异牡荆苷、对香豆酸5 种黄酮类化合物的高效液相色谱的分析方法。通过测定一次蒸煮、二次蒸煮、不同保存条件等因素对箬叶及粽米中黄酮类化合物含量变化的影响,阐明了不同条件下箬叶中黄酮类化合物在粽子中的迁移规律。结果表明：一次蒸煮后,荭草苷、异荭草苷、牡荆苷、异牡荆苷、对香豆酸5 种黄酮化合物从箬叶迁移到粽米的迁移率为7.41%～45.18%;二次蒸煮时,在120 min内箬叶中5 种黄酮类化合物的迁移率为18.07%～65.94%,特别是在前30 min中,箬叶中黄酮类化合物降低明显,分别占120 min总减少量的51.61%～72.48%,并且二次蒸煮的迁移率大于一次蒸煮。在30 d的保存时间内,箬叶中黄酮类化合物在前15 d减少明显,占总减少量的68.60%～85.50%（除异牡荆苷为0外）;第15天后迁出缓慢;30 d内总迁移率分别为25.90%～64.84%（除异牡荆苷为0外）。随着二次蒸煮时间与保存时间的变化,荭草苷与异荭草苷较迁移缓慢。新鲜保存的方式最有利于异牡荆苷在粽子中迁移,速冻保存最有利于荭草苷、异荭草苷、牡荆苷、对香豆酸的迁移。异牡荆苷在真空保存和速冻保存时短时间内不发生迁移。     

Chemical stability of açai fruit (Euterpe oleracea Mart.) anthocyanins as influenced by naturally occurring and externally added polyphenolic cofactors in model systems

The influence of different classes of naturally occurring and externally added polyphenolic cofactors on the phytochemical and colour stability of anthocyanins in açai fruit (Euterpe oleracea) was investigated. Model systems were based on anthocyanin isolates from açai fruit, rich in cyanidin-3-rutinoside (311 ± 27 mg/l) and cyanidin-3-glucoside (208 ± 18 mg/l), and isolated groups of naturally occurring polyphenolic cofactors in açai fruit (phenolic acids, procyanidins, and flavone-C-glycosides, each adjusted to ∼50 mg/l). Anthocyanin degradation kinetics were assessed as a function of pH (3.0, 3.5, and 4.0) and storage temperature (5, 20 and 30 °C). During storage, anthocyanins experienced pH and temperature-dependent losses, and the half life cyanidin-3-rutinoside (t_1/2 = 2.67–210 days) was consistently longer than cyanidin-3-glucoside (t_1/2 = 1.13–144 days). The presence of flavone-C-glycosides induced significant hyperchromic shifts and enhanced anthocyanin stability at all pH and temperature combinations, while no significant effects were attributed to the presence of phenolic acids or procyanidins. Additional models using externally added cofactors from rooibos tea, also rich in flavone-C-glycosides, resulted in up to 45.5% higher anthocyanin colour and up to 40.7% increased anthocyanin stability compared to uncopigmented anthocyanin isolates and had similar copigmentation effects to a commercial rosemary-based colour enhancer. Results suggest flavone-C-glycosides offer potential for their use as colour enhancers and stabilizing agents in products rich in cyanidin glycosides, particularly açai fruit-containing foods, juice blends, and beverages.     

UPLC and HPLC of caffeoyl esters in wild and cultivated Arctium lappa L.

Analytical methods including ultra-performance liquid chromatography (UPLC) and high-performance liquid chromatography (HPLC) with photodiode array (PDA) detector were developed for the analysis of caffeoylquinic acid derivatives in seeds, leaves and roots of Arctium lappa L. Separation was performed on C₁₈ column utilising 5% (v/v) acetic acid in water and acetonitrile at 330 nm. Both methodologies were validated in terms of linearity, precision, and recovery. The results showed that the major advantages of UPLC, over HPLC were the fast analysis, narrow peaks, high sensitivity, and reduction of solvent consumption. Subsequently the methods were applied for the identification and quantification of chlorogenic acid (5-CQA) and 1,5-dicaffeoylquinic acid (1,5-DCQA) as main compounds in samples. The total phenolic content of samples ranged from 3.93 to 14.13 g of 5-CQA equivalent/100 g dry weight (DW). There was a significant variability from 89 to 571 mg/100 g for 5-CQA and 48 to 486 mg/100 g for 1,5-DCQA in dry material.     

Preparative isolation of triterpenoids from Ganoderma lucidum by counter-current chromatography combined with pH-zone-refining

Ganoderma lucidum is a famous fungus. The triterpenoids are the main bioactive components and exhibit various pharmacological activities. However, the scarcity of the pure triterpenoid has greatly restrained the modern research of G. lucidum. The present paper first describes a convenient method to separate the main triterpenoids from G. lucidum using counter-current chromatography (CCC) technique. Ganoderic acid C6 (38 mg), E (76 mg), F (32 mg) were successfully separated from the crude extract in 1 day with the HPLC purity of above 90% using stepwise CCC. Ganoderic acid G (36 mg), A (64 mg), B (25 mg), D (28 mg) and ganoderenic acid D (11 mg) were separated in 2 days with the HPLC purity of above 90% using a combination of stepwise CCC and pH-zone-refining CCC. The method presented in the paper is much quicker and easier than the previous methods.     

Application of preparative high-speed counter-current chromatography for separation and purification of lignans from Taraxacum mongolicum

Apart from being used as a pharmaceutical, the inflorescences, leaves and roots of Taraxacum mongolicum are processed into different food products. However, only few phytochemical investigations on this plant have been performed. In the present study, a preparative high-speed counter-current chromatography (HSCCC) for the separation and purification of bioactive compounds from T. mongolicum was developed. Two lignans, mongolicumin A and rufescidride, were obtained. The target compounds were finally isolated and purified with a solvent system composed of ethyl acetate–n-butanol–water (2:5:7, v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. By injecting 500 mg of the enriched extract of T. mongolicum, one-step HSCCC procedure yielded 36.7 mg of mongolicumin A and 43.9 mg of rufescidride with the purity of 98.7% and 98.5%, respectively, as determined by high-performance liquid chromatography (HPLC). The chemical structures of the two lignans were confirmed by UV, IR, HRESIMS, 1D and 2D NMR. Among them, mongolicumin A is a new compound, and rufescidride was obtained from genus Taraxacum for the first time. Furthermore, lignans were first isolated and identified from genus Taraxacum.     

Composition of flavonoids and phenolic acids in Glycin tomentella Hayata cultivated in various soils

A high-performance liquid chromatography (HPLC) method was developed to simultaneously separate 12 phenolic acids and 21 flavonoids. The separation was performed with a C₁₈ column and a binary gradient solvent system consisting of methanol and water with 9% glacial acetic acid. The peak resolutions (Rs) were 0.51–12.41 and separation factors (α) were all higher than 1. The method was used to survey these phenolic components in Glycin tomentella Hayata. Diadzein was the major flavonoid in the leaves and roots, and gentisic acid and ferulic acid were the major phenolic acids in the leaves. Amounts of phenolic acids and flavonoids in the leaves from varied soil cultivations were in the order: loam > sand > red loamy sand. Flavonoid amounts in the roots also showed the same trend; however, phenolic acid amounts were low and did not present significant differences. Overall, the roots had much higher phenolic contents than the leaves.     

Elucidation of the flavonoid and furocoumarin composition and radical-scavenging activity of green and ripe chinotto (Citrus myrtifolia Raf.) fruit tissues,leaves and seeds

Citrus myrtifolia Raf. (chinotto) flavedo, albedo and carpel membranes from green and ripe fruits, as well as seeds and leaves, were investigated for the first time for their flavonoid and furocoumarin composition. Twenty-three unique compounds distributed in the different fruit/plant parts were identified and quantified. All samples analysed contained flavanone O-glycosides, flavone C- and O-glycosides and furocoumarins; flavedos and albedos also contained significant amounts of polymethoxyflavones. Flavedo and albedo extracts were found to be richest in flavonoids and furocoumarins, containing up to 1.95 g/kg fresh weight. Flavedo, albedo and carpel membranes from ripe fruits were found to be richer than the corresponding tissues from unripe fruits. The remarkable radical-scavenging activity of all the extracts was tested by DPPH^•, ABTS^•+ and FRAP methods, revealing that (i) they were particularly efficient in quenching ABTS^•+ radical cations (up to 9.8 mM Trolox equivalents), and (ii) flavedo and albedo extracts, on average, showed the highest antioxidant capacity.     

Determination of flavone C-glycosides in tea

An HPLC method for the determination of flavone C-glycosides (FCG) from black tea has been developed. Sample clean-up was accomplished by means of polyamide column chromatography, followed by enzyme hydrolysis of interfering compounds such was flavonol glycosides and a second polyamide column Chromatographic step. Using HPLC with gradient elution and photodiode array detection eight FCG were separated. Seven FCG were isolated by means of preparative HPLC. Identification was carried out using co-chromatography, FAB(Fast Atom Bombardment)-mass spectrometry and various nuclear magnetic resonance techniques. Apigenin 6-C-glucosyl-8-arabinoside (schaftoside) and apigenin 6-C-arabinosyl-8-C-glucoside (isoschafto-side) as well as luteolin 8-C-glucoside (orientin) and luteolin 6-C-glucoside (isoorientin) have been detected in tea for the first time. Three of the other compounds have been identified as apigenin 8-C-glucoside (vitexin), apigenin 6-C-glucoside (isovitexin) and apigenin 6,8-di-C-glucoside (vicenin-2). Their occurrence in tea has been previously reported. From its UV spectrum another compound was concluded to be an apigenin glycoside. The FCG were quantified in a variety of teas of different origins (16 black, two green and one oolong). The total amounts of the FCG were 0.48–2.69 g/kg dry weight. The FCG pattern of teas of different origins were similar to each other and no origin-dependent characteristics have yet been observed. Small amounts of FCG (1.2–2.2 mg/g) were detected in hydrolysates of high relative molecular mass fractions (Mr>5000) of a black tea liquor.     

Flavonoid glycosides in the shoot system of Okinawa Taumu (Colocasia esculenta S.)

Six C-glycosylflavonoids and one O-glycosylflavonoid have been isolated from the shoot system of Taumu (Colocasia esculenta S.). They were identified as schaftoside, isoschaftoside, orientin, isovitexin, isoorientin, vitexin and luteolin 7-O-sophoroside. The presence of catechol moiety in the B-ring of isoorientin, orientin and luteolin 7-O-sophoroside showed strong antioxidant activity with different mechanisms of action in DPPH radical scavenging, β-carotene bleaching and superoxide radical inhibition assays. The amount of dry weight matter extract of water was higher than methanol for both the leaf and the stem parts. Isovitexin was the main compound in water and methanol extracts of the leaf while schaftoside was the main compound of the water extract of the stem. Methanol extract of the leaf showed higher DPPH radical scavenging activity than water extract while the opposite was observed for the stem. The results of this experiment suggest the potential of the leaf of Taumu as a source of dietary antioxidant.     

Composition and availability of soluble and insoluble oxalates in raw and cooked taro (Colocasia esculenta var. Schott) leaves

Taro (Colocasia esculenta var. Schott) is a major staple food crop in parts of Asia and the Pacific Islands and is grown as a minor crop in New Zealand. Soluble, insoluble and total oxalate content of young and older leaves were determined by HPLC following hot water (80 °C) and hot (80 °C) acid (0.2 mol/L HCL) extractions. Young taro leaves contained 589 ± 35.8 mg total oxalates/100 g fresh weight (FW) while older taro leaves contained (443 ± 15.0 mg total oxalates/100 g FW). Soluble oxalates were 74% of the total oxalate content of the young and old leaves.     

Isolation and purification of phenylethanoid glycosides from Cistanche deserticola by high-speed counter-current chromatography

Five phenylethanoid glycosides (PhGs), echinacoside, cistanoside A, acteoside, isoacteoside and 2′-acetylacteoside, were isolated and purified from Cistanche deserticola for the first time by high-speed counter-current chromatography (HSCCC) using two biphasic systems, one consisting of ethyl acetate–ethanol–water (5:0.5:4.5, v/v/v) and another of ethyl acetate–n-butanol–ethanol–water (0.5:0.5:0.1:1, v/v/v/v). A total of 28.5 mg of echinacoside, 18.4 mg of cistanoside A, 14.6 mg of acteoside, 30.1 mg of isoacteoside and 25.2 mg of 2′-acetylacteoside were purified from 1412 mg of the n-butanol extract of C. deserticola, each at over 92.5% purity as determined by HPLC. The structures were identified by their retention time, UV, LC–ESI-MS in the negative ion mode, and confirmed by NMR experiments. The characteristic LC–ESI-MSⁿ fragmentation pattern of the five compounds is discussed, and found to be a very specific and useful tool for the structural identification of PhGs from this important medicinal plant.     

Comparison of phenolic compositions between common and tartary buckwheat (Fagopyrum) sprouts

The phenolic compositions of non-germinated/germinated seeds and seed sprouts (at 6–10 day-old) of common (Fagopyrum esculentum Möench) and tartary (Fagopyrum tataricum Gaertn.) buckwheats were investigated. Phenolic compounds, including chlorogenic acid, four C-glycosylflavones (orientin, isoorientin vitexin, isovitexin), rutin and quercetin, were determined in the seed sprouts by high-performance liquid chromatography (HPLC). In the edible parts of common buckwheat sprouts, individual phenolics significantly increased during sprout growth from 6 to 10 days after sowing (DAS), whereas in tartary buckwheat sprouts they did not. While the sum contents of phenolic compounds in the edible part (mean 24.4 mg/g DW at 6–10 DAS) of tartary buckwheat sprouts were similar to those of common buckwheat sprouts, rutin contents in the non-germinated/germinated seeds (mean 14.7 mg/g DW) and edible parts (mean 21.8 mg/g DW) of tartary buckwheat were 49- and 5-fold, respectively, higher than those of common buckwheat. Extracts of the edible parts of both species showed very similar free radical-scavenging activities (mean 1.7 μmol trolox eq/g DW), suggesting that the overall antioxidative activity might be affected by the combination of identified phenolics and unidentified (minor) components. Therefore, buckwheat seed sprouts are recommended for their high antioxidative activity, as well as being an excellent dietary source of phenolic compounds, particularly tartary buckwheat sprouts, being rich in rutin.