Flavonols (kaempeferol,quercetin, myricetin) contents of selected fruits,vegetables and medicinal plants

The concentrations of flavonols (kaempeferol, quercetin, myricetin) were determined in 22 plant materials (9 vegetables, 5 fruits, and 8 medicinal plant organs). The materials were extracted with acidified methanol (methanol/HCl, 100:1, v/v) and analyzed by reverse phase high-performance liquid chromatographic (RP-HPLC) with UV detection. The total flavonols contents varied significantly (P < 0.05) among vegetables, fruits and medicinal plant organs ranged from 0 to 1720.5, 459.9 to 3575.4, and 2.42 to 6125.6 mg kg⁻¹ of dry matter, respectively. Among vegetables, spinach and cauliflower exhibited the highest amounts of flavonols (1720.5 and 1603.9 mg kg⁻¹, respectively), however, no flavonols were detected in garlic. Within fruits, highest level of flavonols was observed in strawberry (3575.4 mg kg⁻¹), whereas, the lowest in apple fruit (459.9 mg kg⁻¹). Of the medicinal plant organs, moringa and aloe vera leaves contained the highest contents of flavonols (6125.6 and 1636.04 mg kg⁻¹), respectively, whereas, lowest was present in barks (2.42–274.07 mg kg⁻¹). Overall, leafy green vegetables, soft fruits and medicinal plant leaves exhibited higher levels of flavonols.     

Determination of glucosinolates and phenolic compounds in rocket salad by HPLC-DAD–MS: Evaluation of Eruca sativa Mill. and Diplotaxis tenuifolia L. genetic resources

The glucosinolate and phenolic profiles of 37 rocket salad accessions (32 Eruca sativa and 5 Diplotaxis tenuifolia) were obtained by liquid chromatography–mass spectrometry. Eleven desulpho-glucosinolates (DS-GLSs) were isolated and the glucosinolate profiles did not differ between the two species. Total DS-GLS content, expressed as sinigrin equivalents (SE) revealed a certain variability, ranging from 0.76 to 2.46 g kg⁻¹ d.w. but, again, the quantitative analysis did not discriminates Eruca from Diplotaxis. The polyphenol evaluation by HPLC-DAD–MS allowed the identification of two different classes of compounds in the two rocket salad species. Qualitative differences were observed between the polyphenol profiles at specific level: quercetin derivatives were the main phenolics of Diplotaxis, whereas kaempferol derivatives characterised Eruca samples. The contents of total flavonoids determined as rutin equivalents (RE) ranged from 4.68 to 31.39 g kg⁻¹ d.w. Kaempferol-3,4′-diglucoside (71.4–82.2%) and isorhamnetin-3,4′-di-glucoside (7.8–18.4%) were always isolated as first and second more abundant phenolic compounds in Eruca samples. No marker phenolic compounds were isolated in Diplotaxis samples.     

HPLC–MS identification and quantification of flavonol glycosides in 28 wild and cultivated berry species

Berries and red fruits are rich dietary sources of polyphenols with reported health benefits. More than 50 different flavonols (glycosides of quercetin, myricetin, kaempferol, isorhamnetin, syringetin and laricitrin) have been detected and quantified with HPLC–MSⁿ in fruits of blueberry, bilberry, cranberry, lingonberry, eastern shadbush, Japanese wineberry, black mulberry, chokeberry, red, black and white currants, jostaberry, red and white gooseberry, hardy kiwifruit, goji berry, rowan, dog rose, Chinese and midland hawthorn, wild and cultivated species of blackberry, raspberry, strawberry and elderberry. The phenolic constituents and contents varied considerably among the analyzed berry species. Elderberry contained the highest amount of total flavonols (450–568 mg kg⁻¹ FW), followed by berry species, containing more than 200 mg kg⁻¹ FW of total: chokeberry (267 mg kg⁻¹), eastern shadbush (261 mg kg⁻¹), wild grown blackberry (260 mg kg⁻¹), rowanberry (232 mg kg⁻¹), american cranberry (213 mg kg⁻¹) and blackcurrants (204 mg kg⁻¹). Strawberry (10.5 mg kg⁻¹) and white currants (4.5 mg kg⁻¹) contained the lowest amount of total flavonols. Quercetins represent the highest percentage (46–100%) among flavonols in most analyzed berries. In wild strawberry and gooseberry the prevailing flavonols belong to the group of isorhamnetins (50–62%) and kaempferols, which represent the major part of flavonols in currants (49–66%). Myricetin glycosides could only be detected in chokeberry, rowanberry and species from the Grossulariaceae, and Adoxaceae family and Vaccinium genus. Wild strawberry and blackberry contained from 3- to 5-fold higher total flavonols than the cultivated one.     

The effect of solar radiation on the flavonol content in broccoli inflorescence

The effect of solar radiation on the quercetin and kaempferol contents in the inflorescence of three broccoli cultivars (‘Lord’, ‘Marathon’ and ‘Fiesta’) was investigated from 1999 to 2001. Great differences in the contents of both flavonols, dependent on growing time and cultivar, were found. Quercetin and kaempferol contents varied from 14.3 to 81.0 mg kg⁻¹ f.w. and from 35.9 to 213 mg kg⁻¹ f.w., respectively. Inflorescences of the cultivar ‘Lord’ were characterised by the highest mean content of quercetin and those of cultivar ‘Fiesta’ of kaempferol. The contents of both flavonols were highly positively correlated with total solar radiation in the period from planting to the harvest of broccoli inflorescences.     

Trace element content of Boletus tomentipes mushroom collected from Yunnan,China

The contents of As, Cd, Cr, Cu, Fe, Hg, Mg, Mn, Ni, Pb, and Zn in eleven fruiting bodies of Boletus tomentipes were determined. The results showed the values of the studied elements decreased in the order: Mg (208–279 mg kg⁻¹) > Fe (106–137 mg kg⁻¹) > Mn (29.5–46.8. mg kg⁻¹) > Zn (18.7–23.1 mg kg⁻¹), > Cu (11.4–15.8 mg kg⁻¹) > Cr (3.36–4.78 mg kg⁻¹) > Pb (1.38–3.88 mg kg⁻¹) > Ni (1.68–3.01 mg kg⁻¹) > Cd (0.16–0.32 mg kg⁻¹) > As (0.10–0.24 mg kg⁻¹) > Hg (<0.06 mg kg⁻¹).     

Preliminary evaluation of nutraceutical and therapeutic potential of raw Spondias pinnata K., an exotic fruit of India

The underutilized, edible green raw (whole) fruits of amra, Spondias pinnata K. (Anacardiaceae family) from the eastern region of India were investigated for their nutraceutical and therapeutic potential. A thorough nutritional characterization of this fruit demonstrated it as a source of energy (348 kcal/100 g DM), phenolic compounds, natural antioxidants and minerals. It is also a moderate source of ascorbic acid, malic acid, calcium, phosphorus and other nutrients. The phytochemical screening revealed 5.12 ± 0.32 mg 100 g^− 1 DM of alkaloids followed by saponins and tannins. All assays were carried out in three different solvent extracts of the fruit. Total phenolic, flavonoid and flavonol contents were obtained as 210 ± 1.89 mg GAE, 28.0 ± 0.91 mg CE and 9.97 ± 0.72 mg RE respectively in 100 mg mix solvent extract (MSE). Antioxidant activity of different extracts (as DPPH scavenging) ranged from 0.73 to 0.59 mg ml^− 1 as IC₅₀ value, ABTS with a trolox equivalent antioxidant concentration (TEAC) value as 0.68 to 0.83 and FRAP 5.97 to 7.93 mg TE 100 mg^− 1 extract. LC- MS/MS analysis revealed the presence of gallic acid, salicylic acid, chlorogenic acid, ellagic acid, p-coumaric acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, quercetin, catechin, myrecetin and rutin. The MSE showed the highest antimicrobial activity (against Listeria monocytogenes (MIC 1.8 mg ml^− 1)) and ??-amylase inhibition capacity (as IC₅₀: 29.3 mg ml^− 1 extract). GC/MS screening showed the presence of vitamin E, furfural, phytosterol, campesterol and fatty acids. Analysis of volatile flavor showed isopropyl myristinate as a major compound followed by the other monoterpenes and sesquiterpenes. The current study explains the nutritional as well as medicinal utility of the fruit which is a rich source of minerals and antioxidants such as phenols and flavonoids.     

Simultaneous determination of bisphenol A,octylphenol, and nonylphenol by pressurised liquid extraction and liquid chromatography–tandem mass spectrometry in powdered milk and infant formulas

A new analytical method, using pressurised liquid extraction (PLE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS), was developed for the simultaneous determination of bisphenol A (BPA), octylphenol (OP) and nonylphenol (NP) in powdered infant formulas (IF) and powdered skimmed milk (PM). The analytes were extracted by PLE, using this optimised conditions: ethyl acetate as solvent, 70 °C of temperature, reversed-phase silica C₁₈ as dispersing agent and three cycles of extraction. The extracts were then injected in LC–MS/MS using a Gemini C₁₈ column and a mixture of 5% water and 95% methanol/acetonitrile, both with 0.1% ammonia, as a mobile phase. Recoveries at different fortification levels (0.5 and 0.05 mg kg⁻¹), were between 89% and 92% for BPA, 84 and 98% for OP, and 93% and 101% for NP. The method was applied to the analysis of samples of PM and IF, bought in Italian and Spanish markets. In positive samples, phenols concentration ranged from 0.07 to 1.29 mg kg⁻¹ for BPA, from 0.028 to 1.55 mg kg⁻¹ for OP and from 0.026 to 1.47 mg kg⁻¹ for NP.     

Occurrence of β-casomorphins 5 and 7 in commercial dairy products and in their digests following in vitro simulated gastro-intestinal digestion

The occurrence of β-casomorphin-5 (BCM5) and β-casomorphin-7 (BCM7) was investigated in commercial dairy products and in their digests, following in vitro simulated gastro-intestinal digestion (SGID), by means of HPLC–MS. The analysed dairy products were as follows: 10 cheeses (Gorgonzola, Caprino, Brie, Taleggio, Gouda, Fontina, Cheddar and Grana Padano 10-, 15- or 25-m ripened); 4 samples of drinking milk (unprocessed, pasteurised, UHT and in bottle-sterilised); 2 yoghurts and 4 fermented milks containing probiotics; 7 infant formulas; and 4 dried milk-derivatives (skim milk powder, calcium caseinate and milk protein concentrates). β-Casomorphin-5 was not detected in dairy products, either prior to or after SGID. β-Casomorphin-7 was detected only in cheeses with the exception of Taleggio, Caprino and Grana Padano samples. Peptide amount was in the range 0.01–0.15 mg kg⁻¹ the highest level being recovered in Brie sample. Following SGID, BCM7 formed in all dairy samples or increased up to 21.77 mg kg⁻¹ in digests of cheeses. The peptide level ranged from 0.29 to 1.23 mg kg⁻¹in fermented milks and from 3.46 to 22.18 mg kg⁻¹ in dried milk-derivatives. Digests of commercial infant formulas contained BCM7 at concentrations of 0.04–0.21 mg l⁻¹. For the first time, this work reports quantitative values for BCM5 and BCM7 in a range of dairy products providing evidence that, during processing, only proteolytic systems involved in manufacturing and ripening of cheese can potentially hydrolyse β-CN to BCM7. Nevertheless, formation or further release of BCM7 is mainly promoted by the action of gastrointestinal proteinases during in vitro digestion irrespective of the type of dairy product.     

Fish and food safety: Determination of formaldehyde in 12 fish species by SPME extraction and GC–MS analysis

The formaldehyde (FA) content in different fish products was evaluated using a solid phase microextraction (SPME)-GC–MS method based on fiber derivatisation with pentafluorobenzyl-hydroxyl-amine hydrochloride. LOD and LOQ values of 17 and 28 μg kg⁻¹, respectively were calculated. Fish quality was assessed by the analysis of 12 species (sea-fish, freshwater-fish and crustaceans), revealing variable FA levels. Fresh, deep frozen, canned, boiled and roasted fish were analysed; cooking always produced a decrease in the analyte content. Fish belonging to the Gadidae family were the samples with the highest FA concentration (from 6.4 ± 1.2 mg kg⁻¹ to 293 ± 26 mg kg⁻¹), in four cases out of 14 exceeding the value of 60 mg kg⁻¹ proposed by the Italian Ministry of Health. Storage on ice was also investigated, showing moderate FA production also at temperature around 0 °C. FA contents lower than 22 mg kg⁻¹ were finally found in all the other samples.     

Field incurred chlorpyrifos and 3,5,6-trichloro-2-pyridinol residues in fresh and processed vegetables

The effect of washing, peeling and cooking on residue levels of chlorpyrifos and 3,5,6-trichloro-2-pyridinol (TCP) in winter (spinach, cauliflower, potato) and summer vegetables (eggplant, tomato, okra) was determined. Analysis was carried out by capillary gas chromatography (DB-5MS capillary column) with mass selective detection. The samples were collected from trials conducted under controlled conditions as well as from the farmer’s field. In supervised field trials, the highest chlorpyrifos residue was found at raw stage in spinach (1.87 mg kg⁻¹) followed by okra (1.41 mg kg⁻¹) and eggplant (1.25 mg kg⁻¹). The lowest residue of chlorpyrifos was recorded in cauliflower (0.036 mg kg⁻¹). The chlorpyrifos residue reduced from 15 to 33% after washing, 65–85% post-peeling and cooking further lowered it from 12% to 48% in all the tested vegetables; while an increase in TCP concentration was observed during heat treatment. Out of 267 vegetable samples collected from the farmer’s field, 225 samples contained detectable residues representing 84% rate of contamination. About 6% of samples contained chlorpyrifos residues above maximum residue limits (MRLs). However, vegetable processing reduced the chlorpyrifos residue below the MRL.     

Investigation on the presence of sulphites in fresh meat preparations: estimation of an allowable maximum limit

Sulphiting agents are commonly used food additives. They are not allowed in fresh meat preparations. In this work, 2250 fresh meat samples were analysed to establish the maximum concentration of sulphites that can be considered as “natural” and therefore be admitted in fresh meat preparations. The analyses were carried out by an optimised Monier-Williams Method and the positive samples confirmed by ion chromatography. Sulphite concentrations higher than the screening method LOQ (10.0 mg · kg^− 1) were found in 100 samples. Concentrations higher than 76.6 mg · kg^− 1, attributable to sulphiting agent addition, were registered in 40 samples. Concentrations lower than 41.3 mg · kg^− 1 were registered in 60 samples. Taking into account the distribution of sulphite concentrations obtained, it is plausible to estimate a maximum allowable limit of 40.0 mg · kg^− 1 (expressed as SO₂). Below this value the samples can be considered as “compliant”.     

Hollow fibre-based liquid phase microextraction combined with high-performance liquid chromatography for the analysis of flavonoids in Echinophora platyloba DC. and Mentha piperita

A simple, inexpensive and efficient three phase hollow fibre liquid phase microextraction (HF-LPME) technique combined with HPLC was used for the simultaneous determination of flavonoids in Echinophora platyloba DC. and Mentha piperita. Different factors affecting the HF-LPME procedure were investigated and optimised. The optimised extraction conditions were as follows: 1-octanol as an organic solvent, pH_donor = 2, pH_acceptor = 9.75, stirring rate of 1000 rpm, extraction time of 80 min, without addition of salt. Under these conditions, the enrichment factors ranged between 146 and 311. The values of intra and inter-day relative standard deviations (RSD) were in the range of 3.18–6.00% and 7.25–11.00%, respectively. The limits of detection (LODs) ranged between 0.5 and 7.0 ng mL⁻¹. Among the investigated flavonoids quercetin was found in E. platyloba DC. and luteolin was found in M. piperita. Concentration of quercetin and luteolin was 0.015 and 0.025 mg g⁻¹ respectively.     

A multi-residue method for fast determination of pesticides in tea by ultra performance liquid chromatography–electrospray tandem mass spectrometry combined with modified QuEChERS sample preparation procedure

A multi-residue method was developed for rapid determination of pesticide residues in tea by ultra performance liquid chromatography–electrospray tandem mass spectrometry (UPLC/MS/MS). The QuEChERS method was used for sample preparation. In order to minimise the matrix effects from tea, a solid phase extraction (SPE) cartridge layered with graphite carbon/aminopropylsilanized silica gel was applied as complementary to QuEChERS method. For accurate quantification, representative matrix-matched calibration curves were applied to compensate matrix effects. Limits of quantification varied with different pesticides but all can be measured at 0.01 mg kg⁻¹ level in a 5 g tea sample except dichlorvos (0.02 mg kg⁻¹). Recoveries ranged from 70% to 120% and relative standard deviation (RSD) met the European United Quality Control guideline. Efficiency and reliability of this method were investigated by the analysis of both fermented and unfermented Chinese tea samples.     

Comparison of the cell wall composition for flesh and skin from five different plums

Cell walls were isolated from flesh and skin of five plum varieties corresponding to three species (Prunus domestica L., Prunus salicina Lindl. and Prunus insititia Lindl.) using the alcohol-insoluble solids (AIS) procedure. Yields varied from 83 to 114 g kg⁻¹ dry weight in the flesh and from 192 to 361 g kg⁻¹ dry weight in the skins. Their main sugars were uronic acid (224–322 mg g⁻¹ AIS), cellulosic glucose (139–170 mg g⁻¹ AIS), galactose and arabinose. Galactose and arabinose ratio were variable between the varieties. The degrees of methylation were high (62–84).     

Phenolic content and antioxidant capacity of tropical highland blackberry (Rubus adenotrichus Schltdl.) during three edible maturity stages

Tropical highland blackberry (Rubus adenotrichus Schltdl.) is a good source of antioxidants and contains appreciable levels of phenolic compounds, mainly ellagitannins and anthocyanins. This study examined the influence of three ripening stages on phenolic contents. Major anthocyanin pigments increased from 0.20 (red fruit) to 1.34 mg g⁻¹ fresh weight (FW) (fully ripe fruit), whereas ellagitannins and ellagic acid derivatives dropped from 3.8 to 2.2 mg ellagic acid equivalents g⁻¹ (FW). Flavonols also dropped from 5.1 to 2.0 mg quercetin equivalents 100 g⁻¹ (FW). Consequently, values for total phenolic compounds ranged from 5.8 to 5.2 mg gallic acid equivalents g⁻¹ (FW), showing no specific trend. Antioxidant activity (H-ORAC) increased from 38.29 to 64.00 μmol of Trolox equivalents g⁻¹ (FW) during ripening. When compared with other commercial cultivars, R. adenotrichus stands out for high H-ORAC value, although comparatively it possesses low anthocyanin content and average total phenolic content.     

Chemical composition of cultivated seaweed Ulva clathrata (Roth) C. Agardh

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5 × 1.5 m tank), or from a large scale system (LSS, 0.8 ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20–26%, essential amino acids accounting for 32–36% of crude protein. The main analysed monosaccharides were rhamnose (36–40%), uronic acids (27–29%), xylose (10–13%) and glucose (10–16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169 mg kg⁻¹ dw) and for Ca (9 vs 19 g kg⁻¹), Fe (0.6 vs 4.2 g kg⁻¹), Cu (44 vs 14 mg kg⁻¹), Zn (93 vs 17 mg kg⁻¹) and As (2 vs 9 mg kg⁻¹). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.     

Chemical characterization and antioxidant evaluation of muscadine grape pomace extract

Concentrated muscadine pomace extract was chromatographically analyzed for its individual phenolic, flavonoid, and anthocyanidin compounds. This extract was also characterized regarding its total phenolic content (TPC), total flavonoid content (TFC), antioxidative activities in terms of scavenging DPPH free radicals, reducing ferric, and chelating Fe²⁺. The TPC of this product was 34.1 ± 1.8 mg of gallic acid equivalents(GAE)/g of extract, and TFC was 3.0 ± 0.3 mg of quercetin equivalents/g of extract. Some phenolic compounds including ellagic acid, gallic acid, (−)-epicatechin, (−)-epigallocatechin, catechin, myricetin, quercetin, and kaempferol were identified. Some anthocyanidins including delphinidin, cyanidin, petunidin, peonidin, and malvidin were also identified in the extract by using a combination of retention and spectral properties on a reverse-phase HPLC–PDA. In addition, 3,3′,4,4′5,5′-hexahydroxystilbene-a resveratrol analogue present in the extract was identified for the first time by LC–MS. The results from this study demonstrate that the muscadine pomace extract is rich of natural antioxidants such as phenolics, flavonoids and anthocyanins, and possesses strong antioxidant properties. Besides, the developed methods can be used for routine quality control of the muscadine products for manufacturing efficiency and consistency.     

Determination of metal contaminations in sea foods from Marmara,Aegean and Mediterranean seas: Twelve fish species

The concentrations of cadmium, cobalt, chromium, copper, iron, manganese, nickel, lead and zinc were determined by ICP-AES in muscles and livers of 12 fish species sampled from the Marmara, Aegean and Mediterranean seas of Turkey. Iron showed the highest levels in examined tissues of all fish species. Following Fe, Zn generally showed the second highest levels. Metal concentrations in edible parts of fish species were 0.02–0.37 mg kg⁻¹ for cadmium, 0.04–0.41 mg kg⁻¹ for cobalt, 0.04–1.75 mg kg⁻¹ for chromium, 0.32–6.48  mg kg⁻¹ for copper, 7.46–40.1 mg kg⁻¹ for iron, 0.10–0.99  mg kg⁻¹ for manganese, 0.02–3.97 mg kg⁻¹ for nickel, 0.33–0.86 mg kg⁻¹ for lead, 4.49–11.2 mg kg⁻¹ for zinc, respectively. All metal concentrations in livers were higher than those in muscles. In some stations, cadmium and chromium concentrations in both muscles and livers, and lead levels in livers of the examined species were higher than permissible safety levels for human uses.     

The effect of diets supplemented with thyme essential oils and rosemary extract on the deterioration of farmed gilthead seabream (Sparus aurata) during storage on ice

The effect on quality were assessed for gilthead seabream fed five different diets: control (basal diet); BHT (basal diet with 200 mg kg⁻¹ of butylated hydroxytoluene); rosemary (basal diet with 600 mg kg⁻¹ of rosemary extract - Rosmarinus officinalis); carvacrol (basal diet with 500 mg kg⁻¹ of essential oil of Thymbra capitata, carvacrol chemotype); and thymol (basal diet with 500 mg g⁻¹ of essential oil of Thymus zygis, subspecies gracilis, thymol chemotype). After 18 weeks of experimentation, the animals were stored on ice at 4 °C for 0, 7, 14, and 21 days. Physical-chemical, microbiological and sensory analyses were carried out at each sampling point to determine the degree of deterioration in the gilthead seabream. Lower indices of oxidation were observed in animals who were administered feeds supplemented with BHT, carvacrol and (to a lesser degree) rosemary. Lower bacteria counts were observed for the BHT and thymol groups, in addition to a slower deterioration in terms of sensory perception. Accordingly, the addition of natural antioxidants to the diet may have an added effect on fish quality, delaying post mortem deterioration.     

Change in anthocyanin concentrations in red apricot fruits during ripening

Here we report on accumulation patterns of anthocyanins and of β-carotene during fruit maturation, between 82 and 125 days after flowering, of two apricot (Prunus armeniaca L.) cultivars, A3576 and A3751. Both cultivars displayed an intense red colour of the skin but differed in their genetic background. The pigments were extracted from skin and flesh, separately, and analysed using HPLC-DAD-MS. Out of three anthocyanins detected here, the major compound, cyanidin-3-O-rutinoside was present at 75%. The two minor compounds were cyanidin-3-O-glucoside and peonidin-3-O-rutinoside. This is the first time that peonidin-3-O-rutinoside has been detected in apricot fruit. During maturation, A3751 accumulated anthocyanins in both skin and flesh, whereas anthocyanins were present only in the skin of A3576. The skin anthocyanin content was higher in A3751 (296 mg kg⁻¹) than in A3576 (41 mg kg⁻¹). Maximum anthocyanin levels were attained after 108 and 118 days of flowering in A3751 and A3576, respectively, in conjunction with loss of firmness and red colour acquisition on the un-blushed side of the fruit. At the end of ripening, the β-carotene flesh concentration reached 5 mg kg⁻¹ in A3576 and 15 mg kg⁻¹ in A3751. A significant effect of environment was observed on the anthocyanin content in the two cultivars.