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.     

Effects of electrode materials on the degradation,spectral characteristics,visual colour,and antioxidant capacity of cyanidin-3-glucoside and cyanidin-3-sophoroside during pulsed electric field (PEF) treatment

The effects of three electrode materials – stainless steel (SS), pure titanium (PT), and titanium-based alloy (TA) – on the PEF-induced degradation, spectral characteristics, visual colour, and antioxidant capacity of cyanidin-3-glucoside (Cy-3-glc) and cyanidin-3-sophoroside (Cy-3-soph) were studied. Cy-3-glc and Cy-3-soph were degraded by PEF; SS retained Cy-3-glc and Cy-3-soph the most, while PT and TA led to greater degradation. Cy-3-glc was more susceptible to PEF than Cy-3-soph. The intensity of UV–Vis spectra absorbance at 520 nm decreased, but it increased at 280 and 325 nm after PEF when using the three electrodes. Their lightness (L^∗) exhibited no changes with SS, but showed a significant reduction with PA and TA. Redness (a^∗) significantly decreased with all three electrodes; the yellowness (b^∗) of Cy-3-glc with PA and TA, and of Cy-3-soph with SS, were also significantly reduced. The antioxidant capacity of Cy-3-glc and Cy-3-soph with SS, in contrast to PT and TA, was enhanced after PEF.     

Interaction between Cyanidin 3-glucoside and Cu(II) ions

The interaction between cyanidin 3-glucoside (Cy 3-glc) and Cu(II) in a binary solvent (methanol–water, 1:1, (v/v)) and in a phosphate buffer was studied. It is demonstrated that Cy 3-glc and Cu(II) ions form a complex that is manifested by appearance of a new absorption band. In the complex, the reduction of Cu(II)–Cu(I) and oxidation of Cy 3-glc takes place. The association constant of the complex formation was calculated using the Rose–Drago method. The complex stoichiometry in the binary solvent was 1:1, and the association constant was K = (28,600 ± 2600) M⁻¹. Interaction of Cy 3-glc and Cu(II) ions has two stages “fast” and “slow”. The kinetics of these processes has been investigated. Thermodynamic data for the “fast” process: enthalpy (ΔH^∗) and entropy (ΔE^∗) of activation, were calculated.     

Physicochemical and antioxidative properties of red and black rice varieties from Thailand,China and Sri Lanka

Nine red and three black rice varieties from Thailand, China and Sri Lanka were analysed to determine their proximate composition and their physicochemical and antioxidant properties. Four groups of rice varieties with different amylose contents were identified. Cyanidin 3-glucoside and peonoidin 3-glucoside were confirmed as the dominant anthocyanins in black rice varieties with contents ranging from 19.4 to 140.8 mg/100 g DM and 11.1–12.8 mg/100 g DM, respectively. Total phenolic content (TPC) differed significantly between the varieties, but not between the colours. Highest TPC was found in the red Thai rice Bahng Gawk (BG) with 691 FA equivalent mg/100 g DM, which showed as well the highest antioxidant properties. In red varieties, the major phenolic acids in the free form were ferulic, protocatechuic and vanillic acid, whereas in black varieties protocatechuic acid was dominant followed by vanillic and ferulic acid. In the bound form, ferulic acid was predominant in both colours, where contents differed significantly, followed by p-coumaric and vanillic acid. The antioxidative capacity did not differ significantly between both colours but amongst genotypes. Antioxidant capacity of rice varieties ranged within 0.9–8.1 mmol Fe(II)/100 g DM for FRAP and 2.1–12.3 mmol TEAC/100 g DM. DPPH scavenging ability ranged from 13.0% to 76.4% remaining DPPH.     

Free and bound total phenolic concentrations,antioxidant capacities,and profiles of proanthocyanidins and anthocyanins in whole grain rice (Oryza sativa L.) of different bran colours

To study the polyphenols in whole grain rice varying in bran colour, the total phenolics, flavonoids and antioxidant capacities of the solvent-extractable (Free) and cell-wall bound (Bound) fractions and the profiles of proanthocyanidins and anthocyanins were determined. Red and purple bran rices had significantly higher total (sum of Free- and Bound-) phenolic (PC) and flavonoid (FC) concentrations and antioxidant capacities than light-coloured bran rice or other cereals (P < 0.05), due to their higher concentrations of proanthocyanidins and anthocyanins, respectively. The concentrations of the Bound-PC and FC accounted for approximately half of the total PC and FC in the light-coloured bran rice, but were lower than those in purple and red bran rice (P < 0.05). High correlations were found between the concentrations of total phenolics and the three antioxidant capacity assays except for those in the bound fraction when related to iron chelating capacity. The concentrations of proanthocyanidins in red bran rice was 1.27 mg/g and its composition was 6.5%, 33.5%, 30.6% and 29.4% of 1–3, 4–6, 7–10 mers, and polymer (>10 mers), respectively. Cyanidin-3-glucoside was the predominant anthocyanin and peonidin-3-glucoside was the second highest; the profiles varied between purple bran cultivars. Whole grain rice differing in bran colour contained unique polyphenol subgroups, which have been proposed to positively impact human health.     

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.     

Anti-oxidative analysis,and identification and quantification of anthocyanin pigments in different coloured rice

Anthocyanin pigments in coloured rice cultivars were isolated and identified using high-performance liquid chromatography techniques. Two black rice cultivars (Asamurasaki, Okunomurasaki) contained three major anthocyanins: cyanidin-3-glucoside, peonidin-3-glucoside and malvidin. Chinakuromai (black) rice additionally contained a fourth anthocyanin, petunidin-3-glucoside. Four red rice cultivars contained only malvidin. The total anthocyanin content varied greatly among black rice cultivars (79.5–473.7 mg/100 g), but was lower in red rice (7.9–34.4 mg/100 g). Total phenolic content was similar between red (460.32–725.69 mg/100 g) and black (417.11–687.24 mg/100 g) rice. The oxygen radical absorbing capacity was ranked as follows: red (69.91–130.32 μmol Trolox/g) > black (55.49–64.85 μmol Trolox/g) > green (35.32 μmol Trolox/g) > white (21.81 μmol Trolox/g) rice. The antioxidant capacity resulted mainly from the seed capsule, not the endosperm. The anthocyanin pigments contributed little to the total antioxidant capacity of red (0.03–0.1%) and black (0.5–2.5%) rice cultivars. Hence, the antioxidant capacity is derived mainly from other phenolic compounds.     

In vitro absorption and antiproliferative activities of monomeric and polymeric anthocyanin fractions from açai fruit (Euterpe oleracea Mart.)

Anthocyanins are among the most important and widely consumed natural pigments in foods, and have attracted increased attention as natural food colourants and potent bioactive agents. However, anthocyanins are generally unstable and may undergo chemical changes that include oxidative and polymerisation reactions during processing and storage. The role of anthocyanin polymerisation reactions on in vitro intestinal absorption and anti-cancer properties has not been assessed. This study investigated the chemical composition, antioxidant properties, antiproliferative activity, and in vitro absorption of monomeric and polymeric anthocyanin fractions from açai fruit (Euterpe oleracea Mart.). Cyanidin-3-rutinoside (58.5 ± 4.6%) and cyanidin-3-glucoside (41.5 ± 1.1%) were the predominant compounds found in monomeric fractions, while a mixture of anthocyanin adducts were found in polymeric fractions and characterised using HPLC–ESI-MSⁿ analyses. Monomeric fractions (0.5–100 μg cyanidin-3-glucoside equivalents/ml) inhibited HT-29 colon cancer cell proliferation by up to 95.2% while polymeric anthocyanin fractions (0.5–100 μg cyanidin-3-glucoside equivalents/ml) induced up to 92.3% inhibition. In vitro absorption trials using Caco-2 intestinal cell monolayers demonstrated that cyanidin-3-glucoside and cyanidin-3-rutinoside were similarly transported from the apical to the basolateral side of the cell monolayers (0.5–4.9% efficiency), while no polymeric anthocyanins were transported following incubation for up to 2 h. The addition of polymeric anthocyanin fractions also decreased monomeric anthocyanin transport by up to 40.3 ± 2.8%. Results from this study provide novel information regarding the relative size, absorption, and bioactive properties of anthocyanin monomers and polymer adducts.     

Effects of baking on cyanidin-3-glucoside content and antioxidant properties of black and yellow soybean crackers

Black soybean is a potential functional food ingredient with high anthocyanin content, but the ability to maintain anthocyanin content under dry heat processing has not been reported. This study investigated the effects of soybean seed coat colour and baking time–temperature combinations on the extractable antioxidant properties of a soy cracker food model. Crackers prepared with black soybeans had significantly higher TPC, total isoflavones, and peroxyl, hydroxyl, and ABTS⁺ radical scavenging abilities than their yellow counterparts, at all time–temperature combinations. Cyanidin-3-glucoside (C3G) was detected only in black soybean crackers, and all baking treatments significantly decreased C3G. The greatest losses occurred at the low temperature × long time and high temperature × short time, the smallest loss with moderate temperature × short/medium time. The high temperature treatment altered phenolic acid and isoflavone profiles; however, total isoflavones were unaffected. Overall results suggest that moderate baking temperature at minimal time may best preserve anthocyanin and other phenolics in baked black soybean crackers.     

Near-infrared oximetry of three post-rigor skeletal muscles for following myoglobin redox forms

We investigated the response of frequency-domain multidistance (FDMD) near-infrared (NIR) tissue oximetry for detecting absolute amounts of myoglobin (Mb) redox forms and their relationship to meat colour stability. Four packaging formats were used to create different blends of Mb redox forms and meat colours during display. Changes in surface colour and subsurface pigment forms during simulated display time (0, 2, 4, and 10 d at 2 °C) were evaluated using surface reflecto-spectrophotometry (both L∗a∗b∗ and specific wavelengths) and FDMD NIR tissue oximetry. Data for both methods of direct measurement of oxymyoglobin and deoxymyoglobin were strongly related and accounted for 86–94% of the display variation in meat colour. Indirect estimates of metmyoglobin ranged from r² = 59–85%. It appears that NIR tissue oximetry has potential as a noninvasive, rapid method for the assessment of meat colour traits and may help improve our understanding of meat colour chemistry in post-rigor skeletal muscle.     

Total carotenoid differences in scallop tissues of Chlamys nobilis (Bivalve: Pectinidae) with regard to gender and shell colour

The aim of this study was to investigate whether total carotenoid content (TCC) in noble scallop Chlamys nobilis is related to body tissue, shell colour, and gender. TCC was determined by a UV–vis recording spectrophotometer in tissue of gonad, adductor, mantle, and gill separately sampled from male and female individuals with orange and brown shell colours from the same cultured population. TCC was significantly different among body tissues, depending on shell colour and gender, ranging from 0.73 to 59.85 μg g⁻¹. In general, TCC was greater in the order of gonad > mantle > adductor > gill. In the same gender, orange shell colour individuals contained significantly higher TCC than brown ones in all four tissues (P < 0.05). In the same shell colour, female contained significantly higher TCC than male in gonad and adductor tissues (P < 0.05).     

Structural and chromatic characterization of a new Malvidin 3-glucoside–vanillyl–catechin pigment

A new pigment, resulting from the reaction between malvidin 3-glucoside and catechin in the presence of vanillin, was detected in a model solution. This newly formed anthocyanin-aryl-flavanol adduct was structurally characterized by 1D and 2D NMR and mass spectrometry, and its chromatic characteristics were studied by UV–Vis techniques. The new pigment was shown to have a wavelength of maximum absorption in the visible region (λ_max) of 549 nm, conferring on it a purple colour, and a molar extinction coefficient value (ε) of 12,247 l · mol⁻¹ · cm⁻¹.     

Rapid determination of free anthocyanins in foodstuffs using high performance liquid chromatography

Anthocyanins constitute a major group of natural pigments, and they are responsible for the colours of fruits and vegetables. A rapid and feasible assay procedure for the determination of free forms of the six most abundant anthocyanins in foods is described. The 3-glucoside forms of pelargonidin, cyanidin, peonidin, delphinidin, petunidin and malvidin with the aglycone cyanidin (as internal standard) were separated by gradient elution and quantified using HPLC-DAD within 18 min. A fast sample preparation step was employed which allows direct injection of samples to the chromatograph without need of chemical extraction. Testing on 28 different vegetable, fruit and processed commercial product samples demonstrated applicability in the concentration range of about 80–420 ng/mL with an accuracy of 99.2 ± 0.2% and an average precision of 0.8%. The method was suggested as a cheap and robust alternative to the previous ones that employ multi-step sample treatment protocols.     

Control of salmon oil photo-oxidation during storage in HPMC packaging film: Influence of film colour

The efforts are being made to design protective hydroxypropyl methylcellulose (HPMC) colour packages to avoid photo-oxidation of edible fats and oils. In the present study, edible films of HPMC containing different edible colours like blue, green, yellow, red and white were tested for their ability to avoid photo-oxidation in salmon oil. The samples taken in petri-dishes and covered with coloured HPMC films of thickness 40 μm were placed under fluorescent light at 20 °C. During storage, chemical parameter of oil quality such as fat oxidation was monitored during 8 days of storage. Oxygen consumption by gas chromatography, conjugated diene values by spectrophotometery and fatty acid composition by gas chromatography (GC) was measured. The results of our study show that HPMC films with suitable edible colours act as adequate light barrier to avoid photo-oxidation of salmon oil during extended storage. HPMC films containing white, red and yellow edible colours show good control of oil photo-oxidation almost similar to the control samples stored in darkness. Oil samples treated with blue and green edible films show gradual increase in oil oxidation with increasing time of light exposure. Oxidation behaviour of samples treated with blue and green films was almost similar to the samples stored in transparent films.     

Antiproliferative effects of cherry juice and wine in Chinese hamster lung fibroblast cells and their phenolic constituents and antioxidant activities

Cherries are good sources of bioactive phenolic compounds that are widely considered to be potentially healthy. Here we investigated the protective activities of juice and wine products of tart and sweet cherries and their constituent anthocyanins (e.g., cyanidin 3-glucoside and cyanidin 3-rutinoside) against oxidative stress induced by hydrogen peroxide (H₂O₂) in Chinese hamster lung fibroblasts (V79-4). Total phenolics in the cherry juices and wines were 56.7–86.8 mg of gallic acid equivalents (GAE)/l and 79.4–149 mg GAE/l, respectively. Total anthocyanins in the cherry juices and wines were 7.9–50.1 mg of cyanidin 3-glucoside equivalents (CGE)/l and 29.6–63.4 mg CGE/l, respectively. Both cherry juices and wines exerted protective effects against oxidative stress induced by H₂O₂ on V79-4 cells and also enhanced the activities of antioxidative enzymes, such as superoxide dismutase and catalase, in a dose-dependent manner. The protection of V79-4 cells from oxidative stress by phenolics was mainly attributable to anthocyanins. The positive correlation between the protective effects against oxidative stress in V79-4 cells and the antioxidant enzyme activities was stronger for cyanidin 3-glucoside than for cyanidin 3-rutinoside.     

Anthocyanin and glucosinolate occurrences in the roots of Chinese red radish (Raphanus sativus L.), and their stability to heat and pH

Roots of three unique Chinese radish cultivars were evaluated as potential sources for anthocyanin-type colourants or value-added products. These cultivars showed high variation in anthocyanins (63.77–160.74 mg/100 g FW). Seventeen pigments were tentatively identified by mass spectroscopy as pelargonidin-3-sophoroside-5-glucoside derivatives with multiple acylation of hydroxycinnamic acids. A bright colour (CIELab) of radish anthocyanins has been shown at a wide pH range, comparably stable at pH < 4.2. Those anthocyanins also showed a remarkable thermal stability, following a zero-order kinetics at pH 2.5 with half-lives of 14.5 or 8.7 h at 90 or 100 °C, respectively. Additionally, those cultivars varied in glucosinolate contents (59.69–163.91 mg/100 g FW), whereas their degradation was sensitive to pH and followed a first-order kinetics at pH 5.8 with half-lives of 11.44 or 7.05 h at 90 or100 °C, respectively. However, the stable pH ranges for anthocyanins and glucosinolates were different: pH < 4.2 and pH > 3.6, respectively. In a radish juice model (pH 5.8/2.5), thermal degradation of anthocyanins or glucosinolates was associated closely with media pH values. In conclusion, cultivar selection, and thermal and pH conditions during processing or storage should be taken into account for quality, stability, and health benefits of radish derived natural colourants or nutraceutical products.     

Phytochemical profile of a Japanese black–purple rice

Black–purple rice is becoming popular with health conscious food consumers. In the present study, the secondary metabolites in dehulled black–purple rice cv. Asamurasaki were analysed using HPLC–PDA–MS². The seeds contained a high concentration of seven anthocyanins (1400 μg/g fresh weight) with cyanidin-3-O-glucoside and peonidin-3-O-glucoside predominating. Five flavonol glycosides, principally quercetin-3-O-glucoside and quercetin-3-O-rutinoside, and flavones were detected at a total concentration of 189 μg/g. The seeds also contained 3.9 μg/g of carotenoids consisting of lutein, zeaxanthin, lycopene and β-carotene. γ-Oryzanol (279 μg/g) was also present as a mixture of 24-methylenecycloartenol ferulate, campesterol ferulate, cycloartenol ferulate and β-sitosterol ferulate. No procyanidins were detected in this variety of black–purple rice. The results demonstrate that the black–purple rice in the dehulled form in which it is consumed by humans contains a rich heterogeneous mixture of phytochemicals which may provide a basis for the potential health benefits, and highlights the possible use of the rice as functional food.     

Flavonoid composition of red sorghum genotypes

The effect of genotype on flavonoid composition was investigated in 13 sorghum varieties using HPLC-DAD. Sorghums with red/purple secondary plant colour had the highest levels of 3-deoxyanthocyanins (32–680 μg/g) with the black pericarp sorghums having the highest. Sorghums with red secondary plant colour had a high proportion of apigeninidin compounds (66–89%), which suggested that secondary plant colour affects 3-deoxyanthocyanin composition. Red pericarp sorghums with tan secondary plant colour had the highest levels of flavones (60–386 μg/g). Flavanones were also detected in all sorghums with a red pericarp (8–48 μg/g) and secondary plant colour did not affect their levels (p > 0.05). The elevated 3-deoxyanthocyanin levels in the black sorghums were due to their pericarp colour. Black sorghum panicles that were exposed to sunlight during their development had three times more 3-deoxyanthocyanins (617 μg/g) than those that were covered with a paper bag (212 μg/g). This study showed that flavonoid levels and composition were affected by sorghum genotype. This information will help sorghum breeders to produce sorghum genotypes with maximum levels of desired flavonoids.     

Anthocyanin and colour changes during processing of pomegranate (Punica granatum L., cv. Hicaznar) juice from sacs and whole fruit

The effects of clarification and pasteurisation on anthocyanins (ACNs) and the colour of pomegranate juice (PJ) produced from sacs and whole fruits were investigated. Clarification caused a loss of 4% of ACNs in juice from sacs (JFS) and a loss of 19% in juice from whole fruit (JFWF). After pasteurisation, there was an 8–14% and 13–9% loss of ACNs from unclarified and clarified JFS and JFWF samples, respectively. Polymeric colour was very high even in unclarified samples (25–29%). Compared to JFS, higher polymeric colour was formed in JFWF. HPLC analyses of PJ revealed that cyanidin-3,5-diglucoside was the major ACN, followed by cyanidin-3-glucoside and delphinidin-3-glucoside. Cyanidin-3,5-diglucoside showed higher stability to clarification and pasteurisation than cyanidin-3-glucoside in both PJ samples. Cold clarification with only gelatin is recommended for PJ. To prevent excessive ACN loss and the formation of brown colouring, PJ should be subjected to minimal heating.