Colour stability of anthocyanins from Isabel grapes (Vitis labrusca L.) in model systems

The influence of temperature, pH, the presence or absence of light, the presence or absence of oxygen and the presence or absence of tannic acid on the colour stability of anthocyanins from crude extract of Isabel grapes (Vitis labrusca L.) in food and model systems was studied. The spectrophotometric results (Δλ, ΔA) revealed interaction between the anthocyanins of the crude extract and tannic acid, suggesting copigmentation. In a model system, temperature, pH, the presence or absence of light, the presence or absence of oxygen and the presence or absence of tannic acid, significantly affected the half-life time of anthocyanins. The long half-time of the anthocyanins from Isabel grapes in a yoghurt model system indicates that these pigments are relatively stable in this food. In a rehydrated beverage model system at 4±1 °C, the presence of tannic acid (1:1, w/v) increased the half-life time by 187 h compared to the control samples.     

Effects of temperature,solid content and pH on the stability of black carrot anthocyanins

Anthocyanin stability of black carrots was studied at various solid contents (11, 30, 45 and 64° Brix) and pHs (4.3 and 6.0) during both heating, at 70–90 °C, and storage at 4–37 °C. Monomeric anthocyanin degradation fitted a first-order reaction model. Degradation of monomeric anthocyanins increased with increasing solid content during heating, while it decreased during storage. For example, at pH 4.3, half-life periods for anthocyanins at 30, 45 and 64° Brix were, respectively, 8.4, 6.9 and 5.2 h during heating at 80 °C and 18.7, 30.8 and 35.9 weeks during storage at 20 °C. At 30–64° Brix, increasing pH from 4.3 to 6.0 enhanced the degradation of anthocyanins during heating. The effect of pH on thermal stability of anthocyanins was also studied at six different pHs (2.5–7.0) in citrate-phosphate buffer solutions and significant decrease in anthocyanin stability was observed at pHs above 5.0. Higher activation energies (E_a) were obtained during heating than during storage with increasing solid contents. At 30–64° Brix, E_a values ranged from 68.8 to 95.1 kJ mol⁻¹ during heating and from 62.1 to 86.2 kJ mol⁻¹ during storage. Q₁₀ values at 20–37 °C were as high as 3.1 at 45° Brix and 3.6 at 64° Brix.     

Determination of the degradation kinetics of anthocyanins in a model juice system using isothermal and non-isothermal methods

The effect of temperature on the degradation of blackcurrant anthocyanins in a model juice system was determined over a temperature range of 4–140 °C. The thermal degradation of anthocyanins followed pseudo first-order kinetics. From 4–100 °C an isothermal method was used to determine the kinetic parameters. In order to mimic the temperature profile in retort systems, a non-isothermal method was applied to determine the kinetic parameters in the model juice over the temperature range 110–140 °C. The results from both isothermal and non-isothermal methods fit well together, indicating that the non-isothermal procedure is a reliable mathematical method to determine the kinetics of anthocyanin degradation. The reaction rate constant (k) increased from 0.16 (±0.01) × 10⁻³ to 9.954 (±0.004) h⁻¹ at 4 and 140 °C, respectively. The temperature dependence of the rate of anthocyanin degradation was modelled by an extension of the Arrhenius equation, which showed a linear increase in the activation energy with temperature.     

外源添加物质对黑莓清汁花色苷稳定性和抗氧化活性的影响

本文研究了咖啡酸、阿魏酸、单宁酸、黄芩素、茶多酚、没食子酸、原儿茶酸、芦丁等8种外源辅色素对黑莓清汁花色苷辅色效果、热稳定性、光稳定性和体外抗氧化活性的影响。结果表明:咖啡酸、原儿茶酸、茶多酚、没食子酸、单宁酸对黑莓清汁花色苷的辅色效果影响较显著(p<0.05),最适浓度为0.1 mmol/L,而芦丁的最佳辅色浓度为0.4 mmol/L;实验所选辅色素能够在一定程度上提高黑莓清汁花色苷的热稳定性,其中没食子酸、阿魏酸、咖啡酸对其影响效果显著(p<0.05),分别使黑莓清汁花色苷的热降解半衰期延长了6.06、2.07和1.39倍。此外,原儿茶酸、阿魏酸、芦丁能够显著提高黑莓清汁花色苷的光稳定性、DPPH自由基清除能力和总还原力(p<0.05);没食子酸和单宁酸的辅色作用降低了黑莓清汁花色苷的光稳定性。黄芩素、阿魏酸、咖啡酸、芦丁等外源物质可作为黑莓清汁花色苷有效的辅色素。     

Copigmentation between malvidin-3-glucoside and some wine constituents and its importance to colour expression in red wine

Thermodynamic parameters for intermolecular copigmentation interactions involving malvidin-3-glucoside were determined by UV/visible spectroscopy at wine pH (pH 3.6). These included association constants, enthalpy and entropy changes, which were measured for chlorogenic acid, caffeic acid, quercetin, quercetin-3-glucoside, (−)-epicatechin, (+)-catechin, procyanidin dimer and seed tannin. Quercetin produced the strongest copigment (K_CP = 2900 ± 1300), whilst the addition of glucose at position 3 (quercetin-3-glucoside) reduced its effect by almost 10-fold. Malvidin-3-glucoside self-association (K_D = 3300 ± 300 mol⁻¹ l) was thermodynamically favoured over intermolecular interaction with any of the copigments tested. No colour enhancement due to self-association was observed for malvidin-3-glucoside-derived pigments that cannot enter hydration reactions. In addition, malvidin-3-(6-O-p-coumaryl)glucoside did not show colour enhancement suggesting that the p-coumaryl group prevents self-association. The malvidin-3-glucoside circular dichroism (CD) spectrum was not affected by indicated changes in malvidin-3-glucoside concentration. These observations demonstrate that self-association of malvidin-3-glucoside is more important than copigmentation in young red wine.     

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.     

Effect of ripeness and postharvest storage on the evolution of colour and anthocyanins in cherries (Prunus avium L.)

The relationship between colour parameters and anthocyanins of four sweet cherry cultivars, Burlat, Saco, Summit and Van was studied. The colour (L^∗, a^∗, b^∗, chroma and hue angle parameters) and anthocyanins were analysed during two different years at two different ripening stages (partially ripe, and ripe, respectively). The cherries were analysed at harvest and after storage at 1.5 ± 0.5 °C and 15 ± 5 °C for 30 and 6 days, respectively. The colour was measured by tristimulus colourimetry (CIELAB system) directly on the fruits, while anthocyanins were quantified by HPLC-DAD analysis on methanolic extracts of freeze-dried samples of the fresh cherries and on the differently stored cherries. L^∗, chroma, and hue angle values were always lower for the ripe than for the partially ripe cherries. All of the cultivars were found to contain cyanidin-3-rutinoside and cyanidin-3-glucoside as the major anthocyanins. The total anthocyanin content in fruits of the different cultivars varied in the order Burlat > Saco > Van > Summit. The concentration of anthocyanins increased at both temperatures of storage in both ripe and partially ripe cherries, but the extent of increase varied among cultivars. Cherries stored at 15 ± 5 °C showed higher reduction of L^∗, chroma and hue angle than fruits stored at 1.5 ± 0.5 °C. L^∗, a^∗, b^∗, chroma and hue angle correlated negatively (P < 0.001) with the total anthocyanins levels, but not with the total phenols. These results show that chromatic functions of chroma and hue correlate closely with the evolution of colour and anthocyanins levels during storage of sweet cherries and indicate that colour measurements can be used to monitor pigment evolution and anthocyanin contents of cherries (and vice versa).     

Colour properties,stability, and free radical scavenging activity of jambolan (Syzygium cumini) fruit anthocyanins in a beverage model system: Natural and copigmented anthocyanins

The colour and stability properties of jambolan anthocyanins, both natural and copigmented forms, were investigated in beverage model as well as their radical scavenging ability. Natural anthocyanins of jambolan revealed low colour intensity due to glycosylation structure of the anthocyanins as diglucoside. The intermolecular copigmentation of anthocyanins with sinapic acid, caffeic acid, ferulic acid, and rosemary polyphenolic extract could enhance the colour intensity, which was observed through spectrometric parameters, such as hyperchromic effect (ΔA_vis-max) and bathochromic shift (Δλ_vis-max). In addition of sinapic acid, caffeic acid, and rosemary polyphenolics also increased the stability of the anthocyanin colour during exposure to white fluorescent light and storage at refrigeration and room temperatures, whereas on high thermal treatments, this phenomenon was not observed. Furthermore, beverage model coloured with natural or copigmented anthocyanins revealed DPPH radical-scavenging activity. The AEAC values indicated that the beverage models with copigmented anthocyanins had higher scavenging activity than the beverage with natural anthocyanins.     

Anthocyanin stability studies in Tibouchina semidecandra L.

The effects of pH, storage period, temperature, light and dark conditions on the stability of anthocyanins extracted from Tibouchina semidecandra flowers of different developmental stages was evaluated. Fully formed but unopened flower bud had the highest amount of total anthocyanin extracted from fresh petals. The anthocyanin contents for all flower developmental stages were stable at pH 0.5–3.0 but the colour of the extracts faded at higher pH values. Degradation percentages of total anthocyanins in the extracts kept at 25 °C were 7–20% lower than that maintained at 31 °C. Extracts stored in darkness at 25 °C maintained their purple colour for 26 days while light exposure reduced it to an average of 10 days. The study shows that suitable storage condition for coloured anthocyanin pigments in extracted form is in acidic conditions in the dark. This implies the potential usage of coloured anthocyanins as natural food colourants and shelf life indicator for acidic foods.     

Spectrophotometric study of the copigmentation of malvidin 3-O-glucoside with p-coumaric,vanillic and syringic acids

Anthocyanins are a natural source of pigments in plants and their processed food products have become attractive and excellent candidates to replace the synthetic colourants due to their characteristic intense colours and associated health benefits. The intermolecular copigmentation between anthocyanins and other colourless compounds has been reported to be an important way to enhance and stabilise the colour intensity of aqueous solutions. In the present work we report the equilibrium constant, stoichiometric ratio and the thermodynamic parameters (ΔG°, ΔH° and ΔS°) related to the intermolecular copigmentation reactions of the anthocyanin malvidin 3-O-glucoside with one hydroxycinnamic acid (p-coumaric acid) and two O-methylated hydroxybenzoic acids (vanillic and syringic acid). Different factors which affect their interactions such as copigment concentration, pH and temperature of the medium are examined at two pH levels (pH = 2.50 and 3.65) corresponding to those of the major food mediums where these reactions take place (fruit juices, wine, jams etc.).     

Petit suisse manufactured with cheese whey retentate and application of betalains and anthocyanins

Petit suisse cheese was elaborated with substitution of 30% milk volume for cheese whey retentate (volumetric reduction ratio=5.0) obtained by ultrafiltration (cheese 1) and 100% milk (cheese 2). These were evaluated regarding physicochemical composition: moisture, ash, total solids, lipids, total proteins, acidity in lactic acid and pH. Natural pigments were added to the cheeses: Cabernet Sauvignon (Vitis vinifera L.) grape anthocyanins or (Beta vulgaris L.) beetroot betalains. The cheese samples were maintained at 6±1 °C for 40 days in light-impermeable packaging and evaluated regarding pigment stability by determining half-life time and percentage color retention. The results of the physicochemical analyses demonstrated that significant differences occurred between cheeses 1 and 2 regarding total solid content, moisture, protein, lipids and carbohydrates. The half-life time and percentage color retention values obtained for the anthocyanin and betalain extracts added to the cheeses were adequate for the shelf life of this product.     

Stability of black carrot anthocyanins in various fruit juices and nectars

Fruit juices (apple, grape, orange, grapefruit, tangerine and lemon) and nectars (apricot, peach and pineapple) were coloured with black carrot juice concentrate and stability of black carrot anthocyanins in these matrices was studied during heating at 70–90 °C and storage at 4–37 °C. Anthocyanin degradation, in all coloured juices and nectars, followed first-order reaction kinetics. During heating, black carrot anthocyanins in apple and grape juices showed higher stability than those in citrus juices at 70 and 80 °C. High stability was also obtained for the anthocyanins in peach and apricot nectars at these temperatures. Black carrot anthocyanins were the least stable in orange juice during both heating and storage. During storage, degradation of anthocyanins was very fast at 37 °C, especially in pineapple nectar. Refrigerated storage (4 °C) markedly increased the stability in all samples. Activation energies for the degradation of black carrot anthocyanins in coloured juices and nectars ranged from 42.1 to 75.8 kJ mol⁻¹ at 70–90 °C and 65.9–94.7 kJ mol⁻¹ at 4–37 °C.     

有机酸对紫甘薯花色苷的辅色作用

辅色作用是一种提高花青素稳定性的有效途径。研究柠檬酸、DL-苹果酸、酒石酸、咖啡酸和阿魏酸对紫甘薯花色苷的辅色作用。结果表明,5 种有机酸均能提高紫甘薯花色苷的吸光度,但均没有改变紫甘薯花色苷的组分。90 ℃条件下,柠檬酸、DL-苹果酸和酒石酸可使紫甘薯花色苷的半衰期由15.3 h分别提高到19.1、19.0 h和16.9 h;咖啡酸与阿魏酸显著地降低了紫甘薯花色苷热稳定半衰期,分别降至1.8 h和1.6 h,不利于紫甘薯花色苷的热稳定性。     

Stabilization of anthocyanin and skullcap flavone complexes – Investigations with computer simulation and experimental methods

We examined the stabilization of anthocyanins with flavones from the practical and theoretical perspective. The influence of addition of skullcap flavones, heating to 50 °C, and 12 day storage time (in the presence and absence of light) on the stability of anthocyanins in honeysuckle concentrates was investigated experimentally. Theoretical study was conducted with molecular dynamics methods in a model system, preceded by simulated annealing and thermalization. By the methods of the computer simulation of the copigmentation process we determined the sites responsible for the stabilization of a cyanidin quinoidal base–baicalin complex. We revealed both direct and water-mediated hydrogen bondings that keep the lamellar stacking structure of these molecules in the bounded form in water medium. The stacking occurs also due to hydrophobic interactions of the rings of both molecules. The experimental part of the study confirmed the effectiveness of anthocyanins stabilization in a concentrate of honeysuckle with the use of skullcap flavones.     

Antioxidant stability of small fruits in postharvest storage at room and refrigerator temperatures

Strawberries (Fragaria ananassa), raspberries (Rubus idaeus) and red currants (Ribes rubrum), as well as two drupes, cherries (Prunus avium), and sour cherries (Prunus cerasus), were subjected to two storage temperatures (4 °C and 25 °C) and phytochemicals concentrations (total phenols, flavonoids and anthocyanins) as well as antioxidant capacity (DPPH, ABTS and FRAP assays) were monitored until the fruit visually spoiled. Red currants and strawberries exhibited the highest initial total phenol (TP) contents (322.40 ± 5.56 and 335.47 ± 6.12 mg GAE/100 g FW, respectively) and maintained the highest TP contents throughout storage at both temperatures. Storage of at 25 °C as opposed to 4 °C, facilitated faster spoilage of analyzed fruits. In addition, most fruits stored at 4 °C, exhibited slightly higher antioxidant activity values at the end of storage according to all three antioxidant activity assays as opposed to fruits stored at 25 °C. The dynamic evolution of antioxidant capacity at both temperatures reflected the transient changes in phytochemical composition of small fruits in storage.     

Physicochemical changes in fresh-cut wax apple (Syzygium samarangenese [Blume] Merrill & L.M. Perry) during storage

Physicochemical changes, such as peel and flesh colours, total anthocyanin content, browning index, firmness, total soluble solid (TSS), titratable acidity (TA), sugar acid ratio (TSS/TA), antioxidant capacity, total phenolic content and ascorbic acid content, in fresh-cut Taaptimjan wax apple fruit stored at 4 ± 2 °C and 12 ± 2 °C for 7 days were investigated. The skin of fresh-cut fruit stored at 4 ± 2 °C showed higher a^∗ value, chroma and total anthocyanin content and lower hue angle than those stored at 12 ± 2 °C. Lightness (L^∗ value) and whiteness index of the fresh-cut fruit flesh stored at 12 ± 2 °C showed significantly lower than those stored at 4 ± 2 °C which related to an significant increase in browning index. Firmness, total soluble solid, titratable acidity and sugar acid ratio did not significant changes during storage. Antioxidant capacity and total phenolic content increased throughout storage. Ascorbic acid content of the fresh-cut fruit stored at 4 ± 2 °C remained constant throughout storage whilst ascorbic content at 12 ± 2 °C decreased and was lower than that at 4 ± 2 °C. At 4 ± 2 °C antioxidant capacity and ascorbic acid content were higher than that stored at 12 ± 2 °C whilst there was no significant difference in total phenolic content. In conclusion, the reduction of whiteness index and the increase in browning index of fresh-cut wax apple flesh were the key factors affecting its quality and storage at 4 ± 2 °C could reduce the change in the flesh colour and maintained the peel colour and nutritional values of fresh-cut wax apple fruit during storage.     

Optimisation of the aqueous extraction conditions of phenols from meadowsweet (Filipendula ulmaria L.) for incorporation into beverages

Meadowsweet was extracted in water at a range of temperatures (60–100 °C), and the total phenols, tannins, quercetin, salicylic acid content and colour were analysed. The extraction of total phenols followed pseudo first-order kinetics, the rate constant (k) increased from 0.09 ± 0.02 min⁻¹ to 0.44 ± 0.09 min⁻¹, as the temperature increased from 60 to 100 °C. An increase in temperature from 60 to 100 °C increased the concentration of total phenols extracted from 39 ± 2 to 63 ± 3 mg g⁻¹ gallic acid equivalents, although it did not significantly affect the proportion of tannin and non-tannin fractions. The extraction of quercetin and salicyclic acid from meadowsweet also followed pseudo first-order kinetics, the rate constant of both compounds increasing with an increase in temperature up until 90 °C. Therefore, the aqueous extraction of meadowsweet at temperatures at or above 90 °C for 15 min yields extracts high in phenols, which may be added to beverages.     

Shelf-life prediction of olive oils using empirical models developed at low and high temperatures

Induction period of the formation of hydroperoxides and conjugated dienes at 50 °C and oxidative stability index at 100–130 °C as the oxidative stability measures of different types of olive oils with a wide range of chemical compositions were determined. Regression models (R² ? 0.95) developed at low and high temperatures showed different contributions of compositional variables (the ratio between monounsaturated and polyunsaturated fatty acids, the content of total tocopherols and phenolics, peroxide value, acid value, and total polar compounds content) to the oxidative stability measures. To estimate the shelf-life of olive oils at low temperature, three empirical models with errors of ±1.5%, <±10%, and ±21.2% were developed.     

Characterisation of sugar beet pectin fractions providing enhanced stability of anthocyanin-based natural blue food colourants

The bathochromic effect resulting from the addition of sugar beet pectin (SBP) to black currant juice, purified anthocyanins (ACN-E) and purified polyphenol extracts comprising both anthocyanins and non-anthocyanin phenolics (PP-E), respectively, were monitored by VIS spectroscopy at varying pH relevant to food formulations (pH 3.6–5.0). Furthermore, a low molecular pectic polysaccharide fraction (PPF) was isolated from SBP using 70% aqueous 2-propanol and further characterised with respect to its chemical composition and mineral content. The marked bathochromic effect was found to be due to the formation of anthocyanin–metal chelate complexes. Only at pH 5.0 SBP entailed a moderate bathochromic shift of 12 nm in model solutions containing ACN-E. This effect was enhanced in PPF model solutions resulting in a λ_max shift up to 50 nm, bringing about an intense blue colour. The bathochromic shift was boosted with increasing pH and upon purification of the anthocyanins. In contrast, after SBP and PPF addition to black currant juice, the bathochromic effect was almost annihilated. Model solutions were stored at 20 °C in the dark for 12 days, and optimum stability of the blue colour was observed at pH 5.0.     

Regulation and improvement of triterpene formation in plant cultured cells of Eriobotrya japonica Lindl

Loquat (Eriobotrya japonica Lindl) is a traditional Chinese medicinal plant that contains triterpenes, which have been shown to exhibit pharmaceutical activities. In this study, we investigated various different culture conditions for cultured cells of loquat to produce triterpenes, including illumination, carbon source, nutrient composition and culture system. When cultured on 2.5 mg/l of 6-benzyladenine, 1 mg/l of naphthalene acetic acid and 30 g/l of sucrose at 25 ± 2 °C in the dark for 30 days, the nutrient composition significantly regulated the cell growth and triterpene production. Supplied with the Murashige and Skoog medium reached higher level of dry weight (1.27 ± 0.09 g per flask) and total triterpene production (151.54 ± 12.58 mg/g of cultured cells), and the N6 medium produced tormentic acid but inhibited other triterpene products, while the B5 medium produced relatively high corosolic acid. Also found, suspension cultures of loquat cell could achieve high productivity as callus culture.