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.     

Effect of γ-irradiation on physical quality attributes and identification properties of different sauces

Gamma-irradiated (0, 1, and 10 kGy) ketchup (KTP), barbeque sauce (BBQ), and sweet chili sauce (SWC) were identified using photostimulated-luminescence (PSL) and thermoluminescence (TL) analysis and characterized for physical properties like Brix, consistency, and color. PSL-based screening showed negative results for all nonirradiated samples, while intermediated results were found for irradiated samples. Routine density separation method for mineral extraction provided better TL results; however, alkaline and acid hydrolysis could ease the process with acceptable results. A slight decrease in consistency of all the samples was observed. Brix of KTP remained unchanged, while increase of same was obvious in BBQ and SWC following irradiation. The Hunter L* value of the γ-irradiated sauces decreased except SWC samples, and the a* and b* values showed a general increase compared to the non-irradiated samples. The studied methods have potential to successfully identify and characterize the irradiated sauces for technical and regulatory requirements.     

Assessment of GH1, CAPN1 and CAST polymorphisms as markers of carcass and meat traits in Bos indicus and Bos taurus–Bos indicus cross beef cattle

The objective of this study was to investigate the effects of bovine GH1, CAPN1 and CAST gene polymorphisms on carcass and meat traits in Nellore and Nellore x Bos taurus beef cattle. Three hundred animals were genotyped for GH1/MspI (TC/G in intron 3), CAPN316 (AF_252504.2:g.5709C > G) and CAST/RsaI (AY_008267.1:g282C > G) and phenotyped for rib eye area, backfat thickness, intramuscular fat, shear force (SF), and myofibrillar fragmentation index (MFI). No significant associations were observed between the GH1/MspI and CAST/RsaI polymorphisms and phenotypes, although the relation between the CAST/RsaI genotypes and meat tenderness evaluated by MFI approached significant. The fact that the CAPN316 polymorphism did not show adequate segregation in Nellore cattle confirms the difficulty of using this marker in breeding programs of different Bos indicus breeds. However, the positive results of the association analysis obtained for Nellore x B. taurus crosses contributed to the validation of previous findings.     

Effects of ultrasonic treatment on the gel properties of microbial transglutaminase crosslinked soy,whey and soy–whey proteins

Food Science and Biotechnology - This paper studied the influences of diverse ultrasonic power treatments on the physico-chemical properties of soy–whey mixed protein induced by microbial...     

GC–MS quantification and sensory thresholds of guaiacol in orange juice and its correlation with Alicyclobacillus spp.

Guaiacol is a profoundly negative taint/off-flavour produced by the increasingly common microbial contamination of fruit juices with Alicyclobacillus spp. The objectives of this study included: determining sensory thresholds for guaiacol in orange juice (OJ), developing an analytical method whose detection limits were equivalent to sensory thresholds and determining levels of Alicyclobacillus spp. that would produce detectable levels of guaiacol. A 12 member trained panel was used to establish guaiacol detection and recognition thresholds. Guaiacol ortho and retro nasal detection thresholds in OJ were 0.70 and 0.53 μg/l respectively. Odour recognition threshold was 2.0 μg/l. A SPME GC–MS Selected Ion Monitoring (SIM) procedure was optimised to achieve analytical detection limits of 0.5 μg/l. Optimum guaiacol detection limit was achieved using only responses from m/z 109 and 124. Ion ratios (m/z 109/124) and linear retention index value matching were used to confirm the identification of guaiacol. Quantification was achieved using peak areas from standard guaiacol additions in orange juice between 0.5 and 100 μg/l. Alicyclobacillus growth of 2.2 log CFU/ml in OJ produced just detectable levels (0.7 μg/l) of guaiacol.     

Identification and distribution of lignans in Punica granatum L. fruit endocarp,pulp, seeds,wood knots and commercial juices by GC–MS

The lignans isolariciresinol, medioresinol, matairesinol, pinoresinol, secoisolariciresinol and syringaresinol were detected and quantified for the first time in pomegranate (Punica granatum L.) and in commercial pomegranate juices by means of GC–MS. The total lignan content in the different plant parts was estimated as follows: 36.1 ± 0.3 μg/g in seeds, 17.8 ± 0.2 μg/g in wood knots, 11.2 ± 0.2 μg/g in fruit pulp, 3.3 ± 0.1 μg/g in endocarp. Secondary metabolite distribution varied greatly in the evaluated samples, with syringaresinol being the most abundant contributor in seeds (23.5 ± 0.4 μg/g) and pinoresinol in knots, pulp, endocarp and juice (8.9 ± 0.3, 7.4 ± 0.2, 3.3 ± 0.1 and 2.1 ± 0.2 μg/g, respectively). A survey on two concentrated juices and three commercial pomegranate beverages evidenced the presence of lignans in all of them, with values ranging from 0.4 ± 0.1 to 4.4 ± 0.1 μg/g. Lignans may be relevant contributors to the purported dietary beneficial properties of pomegranate juice and their considerable abundance in agro-industrial waste materials could render P. granatum by-products an inexpensive and renewable natural source of these healthy compounds.     

Identification and quantification of leaf surface flavonoids in wild-growing populations of Dracocephalum kotschyi by LC–DAD–ESI-MS

Dracocephalum kotschyi Boiss. (Lamiaceae) is an aromatic and perennial herb endemic to Iran with interesting pharmacological and biological properties. The flavonoids luteolin-7-O-glucoside, apigenin-7-O-glucoside (cosmosiin), luteolin 3′-O-β-d-glucuronide, luteolin, apigenin, cirsimaritin, isokaempferide, penduletin, xanthomicrol, calycopterin and the polyphenol rosmarinic acid were identified among 13 natural populations of the plant by ESI–MS, LC–DAD and LC–DAD–ESI-MS. The plant extracts containing the identified compounds showed significant antioxidant activity, which was correlated with the flavonoid content. Additionally, leaf and stem size and geographical variability among the studied populations were correlated with flavonoid accumulation. Canonical correlation analysis was used to find a relationship between plant dimensions and phytochemical composition, and the plants with the lowest growth indices were found to have the highest levels of methoxylated flavonoids.     

Determination of alkylphenol residues in breast and commercial milk by solid-phase extraction and gas chromatography–mass spectrometry

This study presents a feasible and sensitive method to determine alkylphenol residues (i.e., 4-t-octylphenol (4-t-OP) and the isomers of 4-nonylphenols (4-NPs)) in breast milk samples and commercial cow milk products. The matrix interference associated with the constituents in the milk was reduced by extraction with n-hexane and dilution with 50% methanolic solution (v/v, methanol/water), then followed by the Oasis-HLB SPE extraction. The analytes were determined by a GC–MS system in full-scan and selected ion monitoring modes simultaneously. Limit of quantitation was less than 0.05 ng/g in a 20 g (wet weight) milk sample. The 4-NPs were detected in 19 of the 20 breast milk samples at concentrations ranging from 1.7 to 11.6 ng/g, while 4-t-OP was detected in 8 samples at concentrations ranging from 0.4 to 1.1 ng/g. The 4-NPs were detected in all the testing commercial cow milk products at concentrations ranging from 2.9 to 8.8 ng/g.     

Fermentation of commercial soy beverages with lactobacilli and bifidobacteria strains featuring high β-glucosidase activity

An increase of isoflavone aglycone content in soy foods can be attained through fermentation with food-grade bacteria. In this study, two commercial soy beverages with distinctive chemical composition (AS and VS) were fermented by strains of lactobacilli (eight strains) and bifidobacteria (two strains) with a high β-glucosidase activity. Along fermentation, growth of the strains and isoflavone deglycosylation in the soy beverages were monitored. Large differences in growth, aglycone content and chemical parameters in AS and VS beverages fermented by different species and strains were observed. Isoflavone glycosides were completely transformed into their corresponding aglycones by most strains during fermentation of AS beverage, whereas large amounts of undeglycosilated isoflavones were still present in fermented VS. Four strains showing strong deglycosylation activity and appropriate technological properties (Lactobacillus casei LP71, Lactobacillus plantarum E112, Lactobacillus rhamnosus E41 and Bifidobacterium pseudocatenulatum C35) were proposed as industrial starters to improve functionality in soy-based fermented foods.     

Effect of γ-irradiation on the physicochemical and sensory properties of cashew nuts (Anacardium occidentale L.)

The present study evaluated cashew nuts' quality as a function of γ-irradiation dose in order to determine dose levels causing minimal undesirable changes to the product. Physicochemical (colour, peroxide value, hexanal content, fatty acid composition, volatile compounds) and sensory (colour, texture, odor, taste) properties of cashew nuts were determined after irradiation at doses up to 7 kGy. Results showed a five-fold increase in peroxide value (PV) and a two-fold increase in hexanal content after irradiation at a dose of 7 kGy. Of the fatty acids determined, stearic acid concentration increased while oleic acid decreased with irradiation dose. PUFA were unaffected by irradiation. Volatile compounds such as aldehydes, ketones and alcohols increased after irradiation indicating enhanced lipid oxidation. Colour parameter a∗ increased (p < 0.05) at doses >3 kGy while colour parameters L∗ and b∗ remained unaffected by irradiation. Sensory analysis showed that cashew nuts remained organoleptically acceptable at doses <3 kGy.     

Role of β-conglycinin and glycinin subunits in the pH-shifting-induced structural and physicochemical changes of soy protein isolate

Soy β-conglycinin (7S) and glycinin (11S) were incubated up to 4 h in acidic (pH 1.5 to 3.5) or alkaline (pH 10 to 12) solutions to induce protein structural unfolding followed by refolding 1 h at pH 7.0, a process known as pH-shifting. The pH-shifting markedly increased (P < 0.05) emulsifying activity of 11S and to a lesser extent 7S; the former also produced more uniform oil droplets. The emulsifying activity improvements were accompanied by a significant rise in protein surface hydrophobicity, slight loss of the secondary structure (circular dichroism), and substantial dissociation of disulfide-linked basic and acidic 11S subunits. The findings suggested that 11S globulins of soy protein isolate (SPI) were more responsive to pH-shifting treatments than were 7S globulins, and the resulting emulsifying activity enhancements of 11S, in parallel with that of SPI, were indicative of its determinant role in the overall emulsifying properties of pH-shifting-treated SPI. PRACTICAL APPLICATION: Extreme alkaline (pH 12) and acidic (pH 1.5) medium treatments can significantly modify the structure and enhance the emulsifying properties of both β-conglycinin and glycinin components of SPI. The functionality improvement by the pH processes is more remarkable for the glycinin protein fraction. Therefore, SPI enriched with glycinin seems to be particularly suitable for extreme acidic or alkaline processes to produce surface-active functional ingredients for food applications.     

Acidification,crushing and thermal treatments can influence the profile and stability of folate poly-γ-glutamates in broccoli (Brassica oleracea L. var. italica)

The influence of different treatments, i.e., crushing, high temperature short time (90 °C/4 min) (HTST) and low temperature long time (60 °C/40 min) (LTLT) blanching, acidification (pH 4.3), and sequences of these treatments on the folate poly-γ-glutamate profile and stability were investigated. In this study, broccoli was used as a case study. Regarding the folate poly-γ-glutamate profile, endogenous folate poly-γ-glutamates in broccoli florets were found predominantly as hepta- and hexa-γ-glutamates. Crushing raw broccoli, acidification and LTLT blanching enhanced folate deconjugation resulting in monoglutamate, di- and tri-γ-glutamates. Compared to other treatments, HTST blanching preformed prior to crushing resulted in the highest concentration of long chain poly-γ-glutamates. Regarding folate poly-γ-glutamates stability, acidification combined with LTLT blanching decreased folate stability whereas HTST blanching combined with different sequences of blanching and crushing did not affect folate poly-γ-glutamates stability. It was concluded that crushing (prior to heating), acidification and blanching could be strategically applied to increase the folate monoglutamate content of broccoli.     

Fractionation of lipids and purification of γ-linolenic acid (GLA) from Spirulinaplatensis

The microalgae, Spirulinaplatensis, is an excellent source of γ-linolenic acid (GLA), an essential polyunsaturated fatty acid and a potent nutraceutical. The fatty acid composition of S.platensis ARM 740 was determined after transmethylation by gas chromatography (GC). Lipid fractionation was achieved on silica gel column chromatography and preparative TLC. Neutral lipids, glycolipids and phospholipids accounted for 77.0%, 15.6% and 7.4%, respectively, of the total lipid fraction. S.platensis ARM 740 was found to contain 94% of the total GLA in the glycolipid fraction. Attempts were made to purify GLA methyl ester by using urea to form inclusion complexes with the saturated and the less unsaturated FAMEs (fatty acid methyl esters), which enhanced the purity of GLA methyl ester to 84%. A further approach to isolate GLA methyl ester with higher purity involved the use of argentated silica gel chromatography. An initial PUFA concentration step frequently adopted by most researchers to increase GLA purity was not necessary in the isolation of GLA from S.platensis. A GLA methyl ester with a purity of >96% and a recovery of 66% was obtained. Purity of the isolated GLA methyl ester was confirmed by GC and IR analysis with respect to authentic standard.     

Volatiles of the Balkan endemic Daucus guttatus ssp. zahariadii and cultivated and wild-growing D. carota – A comparison study

The hydrodistilled essential oil obtained from the aerial parts of Daucus guttatus Sibth. & Sm. ssp. zahariadii Heywood, an endemic plant species of the Balkan Peninsula, as well as solvent extracts and essential oils from different parts of Daucus carota L. have been analysed by GC and GC–MS and screened for antimicrobial activity against 12 bacterial and two fungal strains. The volatiles of the two plant taxa differed significantly in both their chemical identity and antimicrobial effect. The dominant constituent of D. guttatus oil was apiol (43.3%), which was absent from all samples of D. carota. The diethyl ether extract of D. carota inhibited the growth of the yeast Candida albicans while the oil of D. guttatus at 25 mg/ml had no effect on the growth of the fungal organisms tested. Additionally, the oil of D. guttatus showed prominent antibacterial activity against a pathogenic Corynebacterium pyogenes.     

Isolation of caffeic acid from Perilla frutescens and its role in enhancing γ-glutamylcysteine synthetase activity and glutathione level

Perilla frutescens is an annual herbaceous plant native to Asia, where its leaves are used in Asian gourmet food. Our previous study showed that the inhibition of γ-glutamylcysteine synthetase (γ-GCS) activity was remarkably recovered by pretreatment with perilla leaf extract (PLE). The objective was to fractionise PLE, and to identify the active component that is responsible for the enhancement of γ-GCS activity and glutathione (GSH) concentration. Among the five fractions from PLE, PLE-III of the ethyl acetate fraction showed the highest γ-GCS activity in a HepG2 cell experiment, and was further chromatographed. The purified compound, which enhanced γ-GCS activity, was finally identified as caffeic acid. We first report the enhancement of γ-GCS activity and GSH level in HepG2 cells by caffeic acid obtained from PLE. Our results suggest that caffeic acid may be a key factor in the chemopreventive potential of perilla leaf components by increasing de novo synthesis of GSH.     

cDNA cloning and characterization of Type V/XI procollagen α1 chain in the skate, Raja kenojei

The full-length cDNA of the Type V/XI procollagen α1 [pro-α1(V/XI)] chain was determined by the RACE technique, using a cDNA library constructed from the 4-week embryo of the skate, Raja kenojei. The expression property and phylogenetic analysis confirmed that the skate pro-α1(V/XI) chain was close to other vertebrate pro-α1(V) chains rather than pro-α1(XI) chains. The present study is the first evidence for the primary structure of full-length cDNA of the pro-α1(V/XI) chain in an elasmobranch.     

Comparative study on the physicochemical properties of κ-carrageenan extracted from Kappaphycus alvarezii (doty) doty ex Silva in Tawau,Sabah, Malaysia and commercial κ-carrageenans

κ-Carrageenan is a linear, sulphated polysaccharide that is widely used in the food industry as a gelling agent due to its lack of toxicity and biocompatibility. In this study, the physicochemical properties of κ-carrageenan (TA150) derived from Kappaphycus alvarezii (formerly Eucheuma cottonii) in Tawau, Sabah were investigated and compared to commercial κ-carrageenan (SeaKem CM611, Gelcarin GP812, Gelcarin GP911 NF, and Grindsted^® carrageenan CL220). TA150 exhibited the lowest lightness but highest yellowness, with L^∗ and b^∗ values reported as 82.69 and 17.16, respectively. The rupture strength of κ-carrageenan increased significantly with increasing concentration (p < 0.05). The water losses from κ-carrageenan gel increased with increasing storage times. TA150 lost the most water within 10 days of storage time. The water-holding capacity (WHC) of κ-carrageenan gel was reported to be excellent (>90%) under all storage temperatures (25 °C, 4 °C and −18 °C). The moisture content, ash, acid-insoluble matter, and sulphate levels of κ-carrageenan samples were reported as 3.65–11.41%, 17.75–33.18%, 0.22–3.74%, and 12.00–19.71%, respectively. These samples were low in fat, protein, and crude fibre contents. The potassium content in κ-carrageenan was highest in Gelcarin GP812 (100.42 g/kg), followed by Grindsted^® carrageenan CL220 (61.92 g/kg), TA150 (54.60 g/kg), Gelcarin GP911 NF (40.90 g/kg) and SeaKem CM611 (15.76 g/kg). No heavy metals were detected in TA150 and the other commercial κ-carrageenan samples except for lead. However, the concentration of lead detected in the κ-carrageenan samples fell within the acceptable ranges (<5 mg/kg) set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA).     

Comparison of headspace-SPME-GC–MS and LC–MS for the detection and quantification of coumarin,vanillin, and ethyl vanillin in vanilla extract products

A headspace-solid phase micro-extraction (HS-SPME) GC–MS method has been developed for the determination of coumarin, vanillin and ethyl vanillin in vanilla products. Limits of detection ranged from 1.33 to 13.2 ng mL⁻¹. Accuracy and precision data for the method were measured and compared to those obtained using LC-ESI-MS. A survey of 24 commercially available vanilla products was completed using both techniques. No coumarin was detected in any of the samples. Examination of the GC–MS chromatograms revealed the presence of 18 other flavor related compounds in the samples. The method validation and sample analysis data using HS-SPME-GC–MS were comparable to those obtained using the LC–MS method. Because the two methods are conceptually different from one another, both methods would not be subject to the same interferences. This would allow them to be used as confirmatory methods for each other.