Comparative effect of boiling and solid substrate fermentation using the tempeh fungus (Rhizopus oligosporus) on the flatulence potential of African yambean (Sphenostylis stenocarpa L.) seeds

The tropical African yambean (AYB, Sphenostylis stenocarpa L.) is a protein-rich underutilized African legume. The presence of the flatulence- and diarrhoea-causing raffinose family oligosaccharides (RFO: raffinose, stachyose and verbascose) or α-galactosides has limited the food use of African yambean seeds. To reduce this limitation, non-traditional processing methods are required. Seeds of three varieties were (i) examined for the flatulence- and diarrhoea-causing RFO and (ii) fermented with Rhizopus oligosporus for tempeh production. The traditional tempeh production process involved dehulling, soaking in water for 24 h, boiling in water for 30 min, inoculation and fermentation. In addition, the traditional tempeh procedure was modified by using 1% citric acid solution instead of water for soaking and cooking. Comparisons with traditionally cooked beans, which involved boiling in water for 4 h, were made. Boiling seeds for 4 h resulted in 8–30% reduction of total α-galactosides in the three varieties, while the traditional tempeh procedure resulted in an almost complete loss (98%) of the same (P < 0.05). The modified procedure resulted in a bacteria-free tempeh but α-galactoside reduction was 22–39%. Both tempeh production processes were clearly more effective than was traditional cooking in reducing the flatulence potential of the AYB seeds.     

Aggregation profile, preparation and nutritional characterization of African yam bean (Sphenostylis stenocarpa) acid and salt protein isolates

Aggregation tendencies of African yam bean (Sphenostylis stenocarpa) protein in the various aqueous extraction media with Ca²⁺, Mg²⁺ or at pI (isoelectric point) were determined. Water extractable S. stenocarpa protein aggregates more with addition of either Ca²⁺ or Mg²⁺ than at pI. In alkaline extractants considered (except at pH 10), aggregation tendency of the protein is in the order: pI > Ca²⁺ > Mg²⁺. The protein aggregation trend in salt media is a function of the salt type, aggregation in NaCl solution is of order: Ca²⁺ = pI > Mg²⁺, while in Na₂SO₄, we had pI > Mg²⁺ > Ca²⁺. S. stenocarpa protein aggregation was significantly (P < 0.05) more in Na₂SO₄ than NaCl. The amount of Ca²⁺ required for maximum precipitation of S. stenocarpa from alkaline (water-pH10) extractant was higher than that of Na₂SO₄ and more Ca-proteinate was obtained from the alkaline aliquot. The crude protein of the Ca-proteinates and isoelectric protein isolates obtained from salt and alkaline extract were in the range 71.7–91.8% (dry basis). Protein isolate from alkaline extract had significantly (P < 0.05) higher fat content than the one from salt extract. Isoelectrically precipitated isolates had lower ash content than Ca-proteinates. The percentage ratio of essential to non-essential amino acid was in the range 45–47%. With reference to FAO/WHO standard, the chemical score showed that most of the essential amino acid were in excess, thus, the amino acid distribution of S. stenocarpa protein isolates showed that it can fulfill the essential amino acid requirement of human especially the acid proteins and can be a good protein supplement in food and a wide range of new food products.     

Extractability of African yam bean (Sphenostylis stenocarpa) protein in acid, salt and alkaline aqueous media

Protein extractability from defatted Africa yam been (Sphenostylis stenocarpa) was studied under various conditions: solid/solvent ratio, time, pH, and salt. Extractable protein from S. stenocarpa was strongly dependent on all these factors. Maximum extractable protein was obtained after 2 h extraction time; the solid to solvent ratio in the range of 1:20–1:50 gave maximum protein extractability. The pH corresponding to maximum and minimum extractable proteins were 10 and 5, respectively, but addition of NaCl changed this slightly. Extractable protein of 92%, 88% and 84% were obtained in aqueous concentration of 0.01 MNa₂SO₄, 1.5 MNaCl and 0.01 MNaOH, respectively while other concentrations gave lower extractability. These salt concentrations, i.e. 0.01 MNa₂SO₄ and 1.5 MNaCl gave slightly lower extractable protein under alkaline condition. S. stenocarpa flour has a higher buffer capacity in acid medium than in alkaline medium.     

Rheological characterization of acylated and dextran conjugated African yam bean (Sphenostylis stenocarpa Hochst. Ex A. Rich.) proteins

The molecular weight distribution and rheological properties of acetylated, succinylated and dextran conjugated African yam bean (Sphenostylis stenocarpa) proteins dispersion were studied. Succinylation of the protein showed the three prominent electrophoretic bands of the unmodified protein but all the bands disappeared with acetylation. Immobile band characterized dextran conjugated S. stenocarpa electrophoregram. The flow behavior indices (n) of these modified S. stenocarpa protein dispersions were in the range 0.03–0.22. This is an indication that they were pseudoplastic in nature. This pseudoplastic nature was maintained in ionic media 0.05–0.5 mol dm^?3, pH 3–8 and temperature range of 27–75 °C. The yield stresses were 0.270, 0.302 and 0.320 Pa for acetylated, succinylated and dextran conjugated protein respectively. Activation energy of acetylated and succinylated protein were in the range 6.2–8.2 and 2–5.4 J mol^?1 respectively. Thus acetylation of S. stenocarpa protein made its dispersion viscosity more susceptible to temperature change than succinylation. These results suggest that acylation and dextran conjugation of African yam bean (S. stenocarpa) protein produce protein species with different rheological properties.     

Modification and properties of African yam bean (Sphenostylis stenocarpa Hochst. Ex A. Rich.) Harms starch I: Heat moisture treatments and annealing

African yam bean (Sphenostylis stenocarpa) starch was subjected to heat moisture treatments at 18% (HMT-18), 21% (HMT-21), 24% (HMT-24), 27% (HMT-27) and excess (Annealing) moisture levels. Proximate chemical composition of the starch samples revealed that the moisture content of the starches ranged between 6.7% and 12.5%. Following modification of the native starch, there was a reduction in the moisture level of the heat moisture treated starches from HMT-18 to HMT-27. However, the annealed starch (HMT-ANN) retained higher moisture content compared to native starch (AYB-Native). The carbohydrate, protein, ash, amylose and fat content reduced with all the forms of heat treatments. At the temperature range studied (60–90 °C), increasing level of heat moisture treatments reduced the solubility and swelling capacity. pH also exert a profound effect both on the solubility and the swelling of the starch. Increasing degree of alkalinity increased both solubility and swelling capacity. In the native and modified starch samples, replacement of the wheat flour by the starch resulted in increased alkaline water retention of the blends. Water absorption capacity of the starch increased with the severity of moisture treatments, while the oil absorption capacity decreased. Apart from HMT-18, there was improved gel forming capacity of all the other heat-modified starches.Pasting temperature increased after hydrothermal modifications, whereas peak viscosity (P_v), Hot Paste Viscosity (H_v), setback and breakdown values all reduced after heat moisture treatments. All the starches were of type-B viscosity.Differential scanning calorimetry studies revealed that heat moisture treatment shifted the onset temperature (T_o), peak temperature(T_p) and conclusion temperature (T_c) to higher values. The gelatinisation temperature of the annealed starch was comparable to native starch. In addition, gelatinisation band of the native starch increased progressively from HMT-18 to HMT-27. Heat moisture treatment reduced the gelatinsation enthalpy (ΔH), while the enthalpy of retrogradation(ΔH_r) increased with the storage time of the gelatinised starch. Retrogradation enthalpy of the heat moisture conditioned starches were lower than the value obtained for the native starch.X-ray diffraction studies of the starch indicated that all the starch samples showed the type-C diffraction pattern. Differences were however observed in their degree of crystallinity. Native starch exhibited the lowest crystallinity (20%) while annealed starch had the highest crystallinity (27%)Microscopy studies revealed surface indentation, formation of groves in the central region, folding of starch granules and formation of doughnut-like appearance in some of the starch samples.     

Antioxidant and anti-inflammatory properties of taiwanese yam (Dioscorea japonica Thunb. var. pseudojaponica (Hayata) Yamam.) and its reference compounds

Dioscorea japonica Thunb. var. pseudojaponica (DP) is consumed as food and widely used in traditional Chinese medicine in Taiwan. The aims of this study are to investigate the antioxidant and anti-inflammatory effects of ethanol extract of DP (EDP) and its reference compounds. Fingerprint chromatogram from HPLC indicated that EDP contains gallic acid and vanillic acid. EDP was evaluated for its antioxidant effects and LPS-induced nitrite oxide (NO) production in RAW264.7 cells. EDP decreased the LPS-induced NO production and expressions of iNOS and COX-2 in RAW264.7 cells. In-vivo anti-inflammatory activities of EDP were assessed in mouse paw oedema induced by λ-carrageenan (Carr). We investigated the antioxidant mechanism of EDP via studies of the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) and the levels of malondialdehyde (MDA) in the oedematous paw. The results showed that EDP might be a natural antioxidant and anti-inflammatory agent.     

Rheological properties of Cedrela odorata gum exudate aqueous dispersions

Rheological studies of Cedrela odorata gum aqueous dispersions demonstrated viscoelastic properties. Mechanical spectra derived from small amplitude oscillatory shear (SAOS) results support the onset of gel-like structure at given conditions (10% w/v, 20 °C). The increase in the storage and loss moduli, along with the drop in their frequency dependence, observed at higher gum concentrations, indicates a slower relaxation mechanism, which may be related to the increasing number and structural complexity of transient junction zones among macromolecules. The transition from gel-like to sol-like structure, at higher temperature, was detected from SAOS results. The flow behaviour of the aqueous gum dispersions under steady shear is highly non-Newtonian and can be modelled by the Sisko equation. The Cox–Merz rule was found to fail at critical gum concentration for the onset of gel-like behaviour and also at a temperature slightly above the gel–sol transition. C. odorata gum aqueous dispersions may have interesting applications as stabilisers of emulsions and suspensions on account of their rheological behaviour.     

Deterrent and insecticidal properties of bean seed (Phaseolus vulgaris L.) whole meal or protein extract incorporated into the diet of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae)

Callosobruchus maculatus, a pest that causes serious damage to chickpea Cicer arietinum, cannot develop in the seeds of Phaseolus or Vigna spp. which contain lectins. The insecticidal activity towards C. maculatus in these seeds is attributed both to lectins with specific affinity to N-acetylglucosamine, the major component of insect chitin, and to alpha-amylase inhibitors (lectin-like proteins). The insecticidal properties of bean meal or bean protein extracts from different sources towards different pest species are variable and need to be experimentally evaluated. The main objective of this study was to determine through a feeding trial on artificial chickpea seed the potential of bean seed meal from a wild bean Vigna caracalla, four varieties of Phaseolus vulgaris, and of a protein extract of P. vulgaris seed, to alter different life history traits of C. maculatus. The chickpea weevil was set up on artificial chickpea seeds containing different amounts of bean meal to observe the effects on female oviposition, percentage of development to adulthood and juvenile development time. These traits were combined in a composite index measuring the alteration of the multiplication rate of C. maculatus fed on artificial seed. The activity of lectin-like extracts was observed on chickpea artificial seed spiked with bean seed extract. Incorporation of bean flour at a rate of 10 and 20% in chickpea artificial seed significantly decreased C. maculatus female fecundity, percentage of adult emergence, and greatly increased the development time. Feeding trials with protein extracts of P. vulgaris reduced fecundity and survival of C. maculatus. Incorporation of 10% V. caracalla bean seed meal in chickpea artificial seed, reduced the multiplication potential of C. maculatus by over 90% showing that bean seed lectin extracts are worthy of further investigation for post-harvest infestation control.     

Comparative study of the composition and thermal properties of protein isolates prepared from nine Phaseolus vulgaris legume varieties

Beans contain high amounts of protein and, as demonstrated with other legumes, may contain peptides with bioactive properties. Dozens of dry bean (Phaseolus vulgaris) varieties are grown in Canada, and many vary not only in terms of their protein content but also in their composition. In this study, the composition, molecular structure, and thermal properties of nine varieties of P. vulgaris were studied using electrophoresis, reversed-phase high-performance liquid chromatography (RP-HPLC), differential scanning calorimetry (DSC), and Fourier-transform infrared spectroscopy (FTIR). The predominant protein observed in each variety was vicilin. RP-HPLC chromatogram showed protein isolates (PIs) of white, great northern and black beans to have different profiles of low-molecular-weight proteins. Additionally, black bean PIs were missing proteins in the molecular mass (MM) range of 60 to 97 kDa. DSC analysis showed cranberry and light red kidney bean PIs to have significantly lower denaturation temperatures compared to the other varieties. RP-HPLC chromatograms of cranberry and light red kidney bean PIs were also very different. FTIR spectroscopy showed the predominant secondary structures in all varieties to be ??-sheets and random coils. Increasing the temperature from 25 °C to 95 °C resulted in a reduction in ??-sheet structures and an increase in the content of random coils and antiparallel ??-sheet structures. Overall there were no major dissimilarities observed in the FTIR profiles of the nine P. vulgaris varieties.     

Essential oils of the aerial parts of three Salvia species from Jordan: Salvia lanigera, S. spinosa and S. syriaca

The compositions of the essential oils of three Jordanian Salvia species, S. lanigera and S. spinosa from a desert climate, and S. syriaca from a Mediterranean habitat, were studied. All three oils are rich in monoterpene derivatives (68–73%).     

DNA identification of two laboratory colonies of the weevils, Sitophilus oryzae (L.) and S. zeamais Motschulsky (Coleoptera: Curculionidae) in Taiwan

DNA amplification fingerprinting (DAF) and nuclear ribosomal DNA (nrDNA) amplification were used to discriminate between two laboratory colonies of two closely related species of weevils: the rice weevil, Sitophilus oryzae (L.), and the maize weevil, S. zeamais Motschulsky. For DAF, three sets of primers (aldolase, prolactin receptor, and interleukin-1β) were used for identification by polymerase chain reaction (PCR) and agarose gel electrophoresis, and the highly similar patterns of the resultant amplicons reconfirmed that the two weevils are closely related. The fragments of nrDNA amplification showed that for S. oryzae and S. zeamais, the homologies of the nucleotide sequences of the internal transcribed spacer regions, ITS-1 and ITS-2, were unusually high, at 96% and 97%, respectively. Based on the ITS-1 and ITS-2 sequences, two species-specific primer sets were designed: with the primer set ITS3/So, the predicted 450 bp DNA fragment was yielded with S. oryzae genomic DNA after PCR amplification (n=10), but no PCR product was obtained with S. zeamais (n=10); with the primer set ITS3/Sz, the same 10 S. zeamais specimens yielded a 550 bp DNA fragment, but S. oryzae yielded no amplicons. In view of the difficulty of distinguishing between these two closely related species, the specificity and availability of these two primer sets might prove to be a useful tool for distinguishing between them. However, the nrDNA sequences of the ITS-1 and ITS-2 regions of geographically isolated populations of both weevils still need to be elucidated, and the applicability of this technique to different geographical populations will need to be confirmed by further study.     

Antioxidant potentials and rosmarinic acid levels of the methanolic extracts of Salvia verticillata (L.) subsp. verticillata and S. verticillata (L.) subsp. amasiaca (Freyn & Bornm.) Bornm

This study was designed to examine the in vitro antioxidant activities and rosmarinic acid levels of the methanol extracts of Salvia verticillata subsp. verticillata and S. verticillata subsp. amasiaca. The extracts were screened for their possible antioxidant activity by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid systems. In the first case, S. verticillata subsp. verticillata was superior to the subsp. amasiaca with an IC₅₀ value of 14.5 ± 1.21 μg mg⁻¹. In the β-carotene/linoleic acid test system, inhibition capacity of S. verticillata subsp. verticillata was 74.4 ± 1.29%. Antioxidant activities of BHT, ascorbic acid, curcumin and α-tocopherol were determined in parallel experiments. Activity of rosmarinic acid was also screened for better establishing the relationship between rosmarinic acid level and antioxidant activity for the plant extracts. S. verticillata subsp. verticillata had the highest rosmarinic acid level with a value of 28.7 ± 0.89 μg mg⁻¹. There is a strong correlation between the rosmarinic acid level and antioxidant activity potential. Our results showed that rosmarinic acid and its derivatives are more likely to be responsible for most of the observed antioxidant activities of Salvia species.     

Survival and reproduction of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) following periods of starvation

This study determined the starvation tolerance of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) in terms of both adult survival and reproduction, the impact of starvation on reproduction not having been studied before. Experiments were conducted at 30 °C and 55% or 70% r.h. using a laboratory strain and a field strain of each species. The number of progeny was a better indicator of the impact of starvation on a species than adult survival. Tribolium castaneum was the most tolerant species, requiring up to 35 d starvation before no progeny were produced. Rhyzopertha dominica and S. oryzae required up to 8 d starvation before no progeny were produced. The results suggest that hygiene will have a greater impact on populations of S. oryzae and R. dominica than T. castaneum.     

Anthraquinone profiles, antioxidant and antimicrobial properties of Frangula rupestris (Scop.) Schur and Frangula alnus Mill. bark

Frangula rupestris and Frangula alnus are deciduous shrubs distributed in Balkan Peninsula. While the bark of F. alnus is medicinally widely used, little is known about chemical and biological properties of F. rupestris. In the present study, F. rupestris and F. alnus bark were evaluated for their reducing power, DPPH radical scavenging and chelating activity, as well as antioxidant activity in β-carotene-linoleic acid assay. In addition, phenolic content, anthraquinone profile and antimicrobial activity of F. rupestris and F. alnus bark were determined. The most represented anthraquinone derivatives in F. rupestris and F. alnus bark were physcion (0.11 mg/g) and emodin (2.03 mg/g), respectively. Both species demonstrated excellent antioxidant and antimicrobial activities with MIC values equal or lower than 2.5 mg/mL. The presented research indicates that both species may have health benefits as natural antioxidants and antimicrobial agents for use in functional foods or medicine.     

Inactivation of Escherichia coli, Listeria innocua and Saccharomyces cerevisiae by UV-C light: Study of cell injury by flow cytometry

Flow cytometry (FCM) is a powerful tool for analyzing physiological characteristics of microorganisms on a single-cell basis and identifying heterogeneities within population. This work analyzed the UV-C induced damage on Escherichia coli ATCC 11229; Listeria innocua ATCC 33090 and Saccharomyces cerevisiae KE162 cells by applying flow cytometry technique. The UV-C doses, obtained by altering the exposure time and measured by the iodide-iodate chemical actinometer, ranged between 0 and 5 kJ/m². E. coli; L. innocua and S. cerevisiae populations were quantified by plate count technique. For flow cytometry studies, cells were labeled with fluorescein diacetate (FDA) for detecting membrane integrity and esterase activity, and with propidium iodide (PI) for monitoring membrane integrity. The results showed that mechanisms of cellular damage differed according to time of exposure to ultraviolet radiation and the organism tested. E. coli and S. cerevisiae sub-populations with PI increased within the first minutes of UV-C treatment, without much change afterwards. On the contrary, FCM was used to detect the inactivation of those L. innocua sub-populations of viable microorganisms (maintaining metabolic activity) which were non-culturable due to membrane rupture and thus not detectable by viable plate count technique.     

Nutritional and functional properties of Vicia faba protein isolates and related fractions

The goal of this research was the characterisation of Vicia faba (broadbean) protein isolates and related fractions in order to determine whether this grain legume could be used for production of high quality protein products and other fractions rich in functional components. Alkaline extraction of the defatted seed flour, followed by precipitation at the isoelectric pH, yielded a 92% protein isolate with a high oil absorption capacity. The contents of the favism-inducing glycosides, vicine and convicine, in the isolate were reduced by more than 99% as compared to the original flour, although the amino acid composition was similar to that of the flour. Some of the by-products of protein isolate production may also be of interest from a nutritional and functional point of view. Thus, the oil resulting from hexane extraction of the flour is rich in unsaturated fatty acids, and polyphenols (resulting from extraction of the defatted flour with acetone) showed a high ABTS radical-scavenging activity. In addition, the solid residue (resulting from protein solubilisation) was high in fibre and showed good water absorption. These results show good nutritional and functional properties in V. faba protein isolates and related fractions, which may favour the revalorisation of this traditional bean crop.     

Utilization of galactooligosaccharides by Bifidobacterium longum subsp. infantis isolates

Prebiotics are non-digestible substrates that stimulate the growth of beneficial microbial populations in the intestine, especially Bifidobacterium species. Among them, fructo- and galacto-oligosaccharides are commonly used in the food industry, especially as a supplement for infant formulas. Mechanistic details on the enrichment of bifidobacteria by these prebiotics are important to understand the effects of these dietary interventions. In this study the consumption of galactooligosaccharides was studied for 22 isolates of Bifidobacterium longum subsp. infantis, one of the most representative species in the infant gut microbiota. In general all isolates showed a vigorous growth on these oligosaccharides, but consumption of larger galactooligosaccharides was variable. Bifidobacterium infantis ATCC 15697 has five genes encoding β-galactosidases, and three of them were induced during bacterial growth on commercial galactooligosaccharides. Recombinant β-galactosidases from B. infantis ATCC 15697 displayed different preferences for β-galactosides such as 4′ and 6′-galactobiose, and four β-galactosidases in this strain released monosaccharides from galactooligosaccharides. Finally, we determined the amounts of short chain fatty acids produced by strain ATCC 15697 after growth on different prebiotics. We observed that biomass and product yields of substrate were higher for lactose and galactooligosaccharides, but the amount of acids produced per cell was larger after growth on human milk oligosaccharides. These results provide a molecular basis for galactooligosaccharide consumption in B. infantis, and also represent evidence for physiological differences in the metabolism of prebiotics that might have a differential impact on the host.     

A new variant of antimetabolic protein,arcelin from an Indian bean, Lablab purpureus (Linn.) and its effect on the stored product pest, Callosobruchus maculatus

The anti-metabolic or insecticidal gene, arcelin (Arl) was isolated, cloned and sequenced using sequence specific degenerate primers from the seeds of Lablab purpureus collected from the Western Ghats, Tamil Nadu, India. The L. purpureus arcelin nucleotide sequence was homologous to Arl-3 and Arl-4 alleles from Phaseolus spp. The protein it encodes has 70% amino acid identity with the amino acid sequences of Arl-3I, Arl-3III, Arl-4 precursor, Arl-4 and Arl-4I. The partially purified arcelin from the seeds of L. purpureus using an artificial diet confirmed the complete retardation of development of the stored product pest Callosobruchus maculatus at 0.2% w/w arcelin-incorporated artificial seeds.     

Comparative chemical composition,free radical-scavenging and cytotoxic properties of essential oils of six Stachys species from different regions of the Mediterranean Area

The chemical composition of essential oils of six Stachys species, S. cretica L. ssp. vacillans Rech. fil., S. germanica L., S. hydrophila Boiss., S. nivea Labill., S. palustris L. and S. spinosa L., obtained by hydrodistillation, was studied by GC and GC–MS. All the oils have in common a great percentage of fatty acids and esters (24.2–58.5%) and a high amount of sesquiterpenes (16–35.9%), with the exception of the oil from S. palustris, which consisted mainly of carbonylic compounds (25.4%). The antioxidant activity by DPPH test and the antiproliferative activity on a series of human cancer cell lines (C32, amelanotic melanoma and ACHN, renal cell adenocarcinoma) were investigated for all the oils. S. palustris,S. cretica and S. hydrophila showed the highest antiradical effect, with IC₅₀ values of 0.482, 0.652 and 0.664 mg/ml, respectively. The most antiproliferative essential oil against C32 cell line was the oil of S. germanica with a 77% of inhibition at a concentration of 100 μg/ml. S. germanica, S. palustris and S. spinosa showed the most antiproliferative activity on ACHN cell line, at a concentration of 100 μg/ml,with 81%, 77% and 73% inhibition, respectively.     

Flavonoids of Zizyphus jujuba L. and Zizyphus spina-christi (L.) Willd (Rhamnaceae) fruits

The ripe edible fruits of jujube, Zizyphus jujuba Miller (syn. Z. sativa Gaertner, Z. vulgaris Lam.) and Christ’s thorn jujube Zizyphus spina-christi (L.) Willd (Rhamnaceae family) were phytochemically investigated, comparing their quali-quantitative flavonoids profile. Twelve compounds from both methanol extracts have been recognized as quercetin, kaempferol, and phloretin derivatives by means of HPLC/ESI-MS analyses. Six major compounds have been purified by Sephadex LH-20 column chromatography followed by HPLC and were characterized using NMR spectroscopy. One C-glycoside, 3′,5′-di-C-β-d-glucosylphloretin, was detected in Z. spina-christi. The quantitative analysis of all compounds was also reported showing a higher content of flavonoids in Z. jujuba.