Anthocyanin Peaks Research Articles

Glycosylated flavonoids: fruit's concealed antifungal arsenal.

Fruit defense against pathogens relies on induced and preformed mechanisms. The present contribution evaluated performed resistance of red and green mango fruit against the fungal pathogen Colletotrichum gloeosporioides and identified the main active antifungal components. High-performance liquid chromatography analysis of nonhydrolyzed mango peel extracts identified major anthocyanin peaks of glycosylated cyanidin and methylcyanidin, and flavonol peaks of glycosylated quercetin and kaempferol, which were more abundant on the 'red side' of red mango fruit. Organic extracts of red vs green mango peel were more efficient in inhibiting C.gloeosporioides. Transcriptome analysis of the mango-C.gloeosporioides interaction showed increased expression of glucosidase genes related to both fungal pathogenicity and host defense. Glucosidase treatment of organic peel extract increased its antifungal activity. Additionally, quercetin and cyanidin had significantly higher antifungal activity than their glycosylated derivatives. Peel extract volatiles treated with glucosidase had antifungal activity. GCMS analysis identified 15 volatiles after glucosidase hydrolysis, seven of them present only in red fruit. These results suggest that the fruit obtains a concealed arsenal of glycosylated flavonoids in its peel when they are hydrolyzed by β-glucosidase that is induced in both fungus and host during infection process, become more toxic to the fungal pathogen, inhibiting decay development.

Effects of storage temperature on the bioactive compound content and antioxidant activity of aronia (Aronia melanocarpa) fruit

The fruit of aronia (Aronia melanocarpa, also called the “black chokeberry), which is rich in anthocyanin, polyphenol, and flavonoid content and possesses antioxidant, anticancerous, and anti-inflammatory properties. In this study, the influence of storage temperature and storage period on the phytochemical content and antioxidant activity of aronia was determined. The total polyphenol and flavonoid contents of aronia extract were found to be 308.48 μg gallic acid equivalent/g dry weight and 5.33 μg quercetin equivalent/g dry weight, respectively. HPLC analysis of aronia reveled four anthocyanin peaks corresponding to cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin-3-O-arabinose, and cyanidin-3-O-xylose and three polyphenol peaks corresponding to chlorogenic acid, vanillic acid, and rutin hydrate. Long-term storage at a refrigerated temperature (4℃) reduced the levels the levels of health promoting compounds. We found that the highest amounts of total polyphenols, flavonoids, and anthocyanins were retained in aronia samples stored at -80℃ followed by those stored at -20℃ and 4℃ samples. Furthermore, the samples stored at -80℃ showed the stronger antioxidant activities than those stored at other temperatures. Based on these findings, we concluded that freezing aronia at -80℃ can help preserve its antioxidant activity by maintaining high levels of anthocyanins and other bioactive compounds.

Induction of red skin and improvement of fruit quality in ‘Kent’, 'Shelly' and ‘Maya’ mangoes by preharvest spraying of prohydrojasmon at the orchard

Red-blushed skin color is important for mango fruit quality and marketability. Preharvest spraying of ‘Kent’, 'Shelly' and’ Maya’ mangoes with 0.1, 0.2 or 0.4 % prohydrojasmon (PDJ) induced red skin color (areal coverage and intensity) and anthocyanins accumulation especially in fruit from the outer side of tree canopy, which were exposed to sunlight. Maximum red blush was achieved with 0.2% (‘Shelly’) and 0.4 % (‘Kent’ and ‘Maya’) PDJ in fruit from outside the canopy, with an increase of three fold in anthocyanins and two fold increase in flavonols. HPLC analysis of non-hydrolyzed samples showed that the major anthocyanin peaks were cyanidin-3-galactoside and 7-O-methylcyanidin-3-O-β-D-galactopyranoside, and the major flavonol peaks were glucoside derivatives of quercetin and kaempferol. Thus, PDJ activated the phenylpropanoid and anthocyanin pathways mainly with exposure to sunlight, allowing potential production of specific antioxidant and antifungal flavonols and anthocyanins. Induced red skin color in most of the experiments reduced postharvest decay, which may enhance commercial value and extend shelf life.

β-Cyclodextrin-assisted extraction and green chromatographic analysis of Hibiscus sabdariffa L. anthocyanins and the effects of gallic/ferulic/caffeic acids on their stability in beverages

This study aimed to develop a green approach for extraction and determination of anthocyanins from Hibiscus sabdariffa calyx using a hydroxypropyl-β-cyclodextrin (HPβ-CD)-assisted extraction technique coupled with HPLC-DAD analysis and to investigate the effects of gallic/ferulic/caffeic acids on the stability of H. sabdariffa anthocyanins in beverages. HPβ-CD-assisted extraction of total anthocyanins from H. sabdariffa calyx was optimized using a Box–Behnken design-response surface methodology, and extraction at 54 °C for 53 min using HPβ-CD aqueous solution (3.7 g/100 mL) gave the maximum extraction yield of total anthocyanins (4.82 ± 0.13 mg/g). A work-safe and rapid HPLC-DAD method was developed for analysing H. sabdariffa anthocyanins using ethanol and a tartaric acid aqueous solution as mobile phases, and the main anthocyanin peaks were determined as delphinidin-3-O-sambubioside and cyanidin-3-O-sambubioside using a two-dimensional HPLC-DAD/ESI-MS method. Method validation showed that the developed method was accurate, stable and reliable for the analysis of H. sabdariffa anthocyanins. Additionally, the degradation of delphinidin-3-O-sambubioside, cyanidin-3-O-sambubioside and total anthocyanins from H. sabdariffa calyx ideally followed a first-order model (R2 > 0.98) in beverages alone or with additional phenolic acids. Gallic acid improved the stability of H. sabdariffa anthocyanins better than ferulic and caffeic acids, and adding/increasing ferulic and caffeic acids accelerated anthocyanin degradation.

Pharmacopeial HPLC identification methods are not sufficient to detect adulterations in commercial bilberry (Vaccinium myrtillus) extracts. Anthocyanin profile provides additional clues

Current pharmacopeias provide HPLC anthocyanin profiles to identify commercial bilberry extracts. However, the pharmacopeial identification protocols may not be sufficient enough to distinguish genuine bilberry extracts from adulterated material. This is primarily due to the non-inclusion of literature-reported anthocyanin profile and compositional variations in bilberry when sourced from different geographical regions. Using anthocyanin profiles of both authentic bilberry extracts and literature reports, we attempted to provide appropriate identification protocol for genuine bilberry extracts. We compared HPLC anthocyanin profiles of selected ‘suspected’ adulterant species and adulterant-spiked bilberry extracts to decipher clues to infer adulteration. The clues include appearance of new anthocyanin peaks and changes in compositional ratios of anthocyanins. In addition, we attempted to provide likely adulterants based on ‘economic motivation’ and market place information and appropriate clues to identify them in adulterated commercial bilberry extracts.

Characterization of Anthocyanins in Caucasian Blueberries (Vaccinium arctostaphylos L.) Native to Turkey

High-performance liquid chromatography (HPLC) combined with diode array (DAD) and electrospray ionization mass spectrometric (ESI-MS) detections were used to characterize anthocyanins in the berries of Vaccinium arctostaphylos L. The dark purple-black berries were collected from five Caucasian blueberry populations in northeastern Turkey. The HPLC-DAD profile consisted of 19 anthocyanin peaks, but HPLC-ESI-MS revealed fragment ion patterns of 26 anthocyanins. Delphinidin, cyanidin, petunidin, peonidin, and malvidin were all glycosylated with four different monosaccharide moieties (galactose, glucose, arabinose, and xylose) with the first two also conjugated with rhamnose. Furthermore, anthocyanidin disaccharides, tentatively identified as anthocyanidin sambubiosides, were characteristic for these berries. The mean content of the total anthocyanins was 1420 mg/100 g dry weight. The most predominant anthocyanidins were delphinidin (41%), petunidin (19%), and malvidin (19%). Glucose was the most typical (61%) sugar moiety. This study revealed that wild Caucasian blueberries contain an abundance of bioactive anthocyanins and thus are ideal for various functional food purposes.

Differences in the Floral Anthocyanin Component of Lantana in Taiwan

Lantana (Verbenaceae) has adapted to the natural environment of Taiwan, and is thought to be a naturalized weed. A total of 43 Lantana plants, comprising 1 species of 3 varieties (38 plants collected from 38 sites in Taiwan) and 5 horticultural plants, were collected in Taiwan. In order to study the relationship between wild species and horticultural plants, we used co-HPLC to examine each anthocyanin peak and the content ratio in mature flowers as biochemical characters, along with two morphological characters. As a result of HPLC analysis, five out of six anthocyanin peaks were detected in 42 Lantana plants. Pigments were identified as: 3-glucoside, 3,5-diglucoside, and 3-malonylglucoside of cyanidin, and 3,5-diglucoside and 3-malonylglucoside of peonidin. The anthocyanin ratios in each plant were different; they presented several significant ratios. Based on these results, we can distinguish wild from horticultural plants. The flower color of Lantana will become more diverse through crosses between wild and horticultural plants.

Pressurized solvent extraction and monolithic column-HPLC/DAD analysis of anthocyanins in red cabbage

The aim of this work was to develop a fast method for extraction and analysis of anthocyanins in red cabbage. Pressurized hot water containing 5% of ethanol was used as an extremely efficient extraction solvent. HPLC/DAD with a monolithic column was used to accomplish a fast analysis—24 anthocyanin peaks within 18 min. Statistical design was used to optimize the studied extraction parameters: temperature (80–120 °C); sample amount (1–3 g); extraction time (6–11 min); concentration of formic acid in the extraction solvent (0–5 vol.%). The best extraction conditions for a majority of the anthocyanin peaks were 2.5 g of sample, 99 °C (at 50 bar), 7 min of extraction and a solvent composition of water/ethanol/formic acid (94/5/1, v/v/v).

Development of Analyses by High-Performance Liquid Chromatography and Liquid Chromatography/Tandem Mass Spectrometry of Bilberry (Vaccinium myrtilus) Anthocyanins in Human Plasma and Urine

Anthocyanins are potent antioxidants that may possess chronic disease preventive properties. Here, rapid, reliable, and reproducible solid-phase extraction, high-performance liquid chromatography (HPLC), and mass spectrometry techniques are described for the isolation, separation, and identification of anthocyanins in human plasma and urine. Recoveries of cyanidin-3-glucoside (C3G) were 91% from water, 71% from plasma, and 81% from urine. Intra- and interday variations for C3G extraction were 9 and 9.1% in plasma and 7.1 and 9.1% in urine and were less than 15% for all anthocyanins from a standardized bilberry extract (mirtoselect). Analysis of mirtoselect by HPLC with UV detection produced spectra with 15 peaks compatible with anthocyanin components found in mirtoselect within a total run time of 15 min. Chromatographic analysis of human urine obtained after an oral dose of mirtoselect yielded 19 anthocyanin peaks. Mass spectrometric analysis employing multiple reaction monitoring suggests the presence of unchanged anthocyanins and anthocyanidin glucuronide metabolites.

Anthocyanidins of Sarracenia L. Flowers and Leaves

Additional index words. Sarracenia flava, Sarracenia leucophylla, Sarracenia psitticina, Sarracenia purpurea, Sarracenia rubra, pitcher plant, pigments, anthocyanin quantity (11%) was present in S. psitticina. Sarracenia flava has yellow flowers and contained no anthocyanidins. Romeom et al. (1977) reported that quercetin glycosides were the principal flavonols present in Sarracenia. In the biochemical pathway, dihydroquercetin is a precursor of both quercetin and cyanidin. Therefore, we expected to find cyanidin as the principal anthocyanidin in Sarracenia leaves and flowers. Romeo et al. (1977) found a small amount of kaempferol in Sarracenia, but we did not detect pelargonidin, which is derived from dihydrokaempferol. This also was not unexpected because many Petunia ×hybrida Hort. Vilm. cultivars contain small amounts of kaempferol and produce no pelargonidin (Griesbach and Asen, 1990; Griesbach et al., 1991). The discovery of delphinidin was not unexpected since Schnell (1978) had reported the presence of blue pigments in S. psittacina and S. purpurea. This current study identifies those blue pigments as delphinidin glycosides. Schnell, using TLC analysis, reported the presence of 13 different floral pigments. Our HPLC profiles of the floral anthocyanins of S. rubra and S. leucophylla before hydrolysis showed only a single anthocyanin peak (data not shown). In S. purpurea and S. psitticina, five different anthocyanin peaks were present, two of which were present in only trace amounts (data not shown). Many of the 13 pigments reported by Schnell were probably breakdown products. Further studies are needed to identify the sugar(s) attached to the two major anthocyanidins.

Stability of Anthocyanins in Various Vegetables and Fruits.

Anthocyanins were isolated from 19 kinds of vegetables and fruits. Individual anthocyanins were analyzed by HPLC using a photodiode array detector and were examined for the heating and ultraviolet irradiation stabilities of these anthocyanins. This report deals with the relationship between anthocyanin composition and heating or ultraviolet irradiation stabilities of anthocyanins. Stable anthocyanins showed many anthocyanin peaks and a high ratio of acylated anthocyanin peaks. This result shows that the anthocyanin composition and the rate of acylation affect the stability of anthocyanins.

Storage Changes in Anthocyanin Content of Red Raspberry Juice Concentrate

ABSTRACTAnthocyanins from red raspberry juice concentrates were extracted with acidified ethanol and methanol and separated by high performance liquid chromatography. Six anthocyanin peaks were identified as cyanidin‐3‐sophoroside (cy‐3‐sop), cyanidin‐3‐glucorutinoside (cy‐3‐glru), cyanidin‐3‐glucoside (cy‐3‐gl), cyanidin‐3‐rutinoside (cy‐3‐ru), pelargonidin‐3‐sophoroside (pg‐3‐sop), and pelargonidin‐3‐glu‐corutinoside (pg‐3‐glru). Relative proportions of individual anthocyanins were determined by comparing peak areas. The proportions of cy‐3‐sop, cy‐3‐glru and cy‐3‐gl in particular changed greatly on storage for 3 mo at + 20°C. Minor changes occurred when the raspberry juice concentrate was stored at −20°C.

Effect of Catechin and Acetaldehyde on Colour of Saskatoon Berry Pigments in Aqueous and Alcoholic Solutions

The influence of acetaldehyde and catechin on aqueous solutions of cyanidin 3-glucoside and aqueous and alcohol extracts of saskatoon berry was investigated. Pigment systems were analyzed periodically during storage at 23°C in the dark for absorbance spectra, high performance liquid chromatography (HPLC) profiles and Hunterlab colour values. Presence of acetaldehyde in the fruit extracts and both acetaldehyde and catechin in the cyanidin 3-glucoside model system caused a marked increase in colour intensity during storage. Visible spectra of the colour-intensified samples showed both a bathochromic shift and an increase in absorbance. The HPLC profiles of the acetaldehyde treated berry extracts revealed the appearance of a new peak and the decrease of two anthocyanin peaks. Chromatograms of the cyanidin 3-glucoside solution containing acetaldehyde and catechin displayed six new peaks and loss of the anthocyanin. Addition of catechin alone had no significant effect on the colour of the model or pigment extract systems. Results indicated that colour intensification was due to molecular condensation involving catechin, acetaldehyde and anthocyanin as well as other phenolic compounds in the berry extracts. Hue angle of the extracts accurately measured the colour intensification reaction.