Hydrotropic extraction of bioactive limonin from sour orange (Citrusaurantium L.) seeds

Limonoids are potential bioactive compounds present only in citrus among fruits and vegetables. A new process for extraction of limonoid aglycones from sour orange (Citrus aurantium L.) seeds was investigated using aqueous hydrotropic solutions. The extraction efficiency was dependent on hydrotrope concentration, extraction temperature and percent of raw material loaded. Two hydrotropes such as sodium salicylate (Na-Sal) and sodium cumene sulphonate (Na-CuS) were studied using Box-Behnken experiment design. Response surface analysis (RSA) of data was performed to study the effect of parameters on extraction efficiency. Prominent limonoid aglycone such as limonin was extracted and quantified for process optimization. Both hydrotropes gave maximum limonin yield at 2 M concentration, extraction temperature of 45 °C and 10% solid loading. A maximum limonin yield of 0.65 mg/g seeds was obtained using Na-CuS whereas only 0.46 mg/g seed was obtained using Na-Sal. Using this process, the use of organic solvents can be reduced dramatically to keep the process environmental friendly for the extraction of bioactive compounds.     

Simultaneous quantification of selected compounds from Salvia herbs by HPLC method and their application

A simple and reliable reversed-phase high performance liquid chromatographic method was developed and validated for simultaneous determination of danshensu, protocatechuic aldehyde, rosemarinic acid, salvianolic acid B, dihydrotanshinone I, tanshinone I, cryptotanshinone and tanshinone IIA in Salvia herbs. The contents of these eight main components were compared between Salvia miltiorrhiza Bunge and Salvia przewalskii Maxim, which were used for treating coronary heart diseases, cerebrovascular disease, bone loss, hepatitis, hepatocirrhosis and chronic renal failure. The samples were successfully separated by the HPLC method. All the standard compounds showed a good linearity (R² > 0.9994) in the relatively wide concentration range. The limit of detection of the eight compounds was in the range of 0.05–0.5 μg/ml and the limit of quantification was in the range of 0.25–1.50 μg/ml. The intra-day variability was in the range of 0.03–8.08% and the inter-day variability was in the range of 0.18–12.36%. The recoveries of the selected compounds were in the range of 96.2–108.1%. This method was accurate, precise and reproducible, and could be successfully applied to the quality control and stable experiment for the preparations consisted of these active components, and the content comparison of the herbs from Salvia species.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Subcritical water extraction of phenolic compounds from potato peel

Potato peel, a waste generated from potato processing, is a disposal problem. But, potato peel is a good source of functional ingredients such as phenolic compounds. This study investigated the extraction of eight phenolic compounds (Gallic acid, GAC; Chlorogenic acid, CGA; Caffeic acid, CFA; Protocatechuic acid, PCA; Syringic acid, SGA; p-hydroxyl benzoic acid, PBA; Ferulic acid, FRA and Coumaric acid, CMA) from potato peel using subcritical water.Experiments were performed in a batch stainless steel reactor at 6 MPa, 2 mL/min and temperatures ranging from 100 to 240 °C at residence time of 30 to 120 min. High recoveries of phenolic compounds (81.83 mg/100 g; wet basis wb) were obtained at 180 °C and extraction time of 30 min compared to 3 h extraction with methanol (46.59 mg/100 g wb). CGA (14.59 mg/100 g wb) and GAC (29.56 mg/100 g wb) were the main phenolic compounds obtained from potato peel at 180 °C. Subcritical water at 160 to 180 °C, 6 MPa and 60 min might be a good substitute to organic solvents such as methanol and ethanol to obtain phenolic compounds from potato peel.     

Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

Determination of flavonoids in a Citrus fruit extract by LC–DAD and LC–MS

Flavonoids are a group of polyphenolic compounds with health-related properties. Citrus fruits are rich in flavonoids and their extracts are being used as functional ingredients for several industrial products. A new high performance liquid chromatography technique with an UV photodiode-array detector was used to analyze flavonoids of an extract of Citrus species. To our knowledge this is the first study that reports isoquercitrin presence at a level of 77.3 mg/100 g in a sample made of Citrus fruits; four other flavonoids were quantified as rutin (326.59 mg/100 g), naringin (338.36 mg/100 g), quercetin (96.35 mg/100 g) and naringenin (2.35 mg/100 g). Identification was confirmed by a liquid chromatography mass spectrometer system. Method validation was achieved, providing an analytical technique that can be used to detect trace amounts of these compounds in Citrus extracts with an extremely rapid sample preparation.     

Identification and quantification of phenols, carotenoids, and vitamin C from papaya (Carica papaya L., cv. Maradol) fruit determined by HPLC-DAD-MS/MS-ESI

Recent studies have demonstrated that vitamin C, phenols, and carotenoids are bioactive compounds that protect the body from oxidative stress, reducing the risk of cardiovascular diseases and some types of cancer. Qualitative and quantitative analysis of the major phytochemicals found in papaya fruit flesh and skin (Carica papaya L., cv Maradol) was conducted in four stages of ripeness, using high-performance liquid chromatography mass spectrometry. Phenolic compounds identified in the fruit skin tended to decrease with ripening. The compounds identified were ferulic acid (277.49 to 186.63 mg/100 gDW), p-coumaric acid (229.59 to 135.64 mg/100 gDW), and caffeic acid (175.51 to 112.89 mg/100 gDW). The following carotenoids, along with vitamin C, increased in flesh with ripening; lycopene (0.36 to 3.40 mg/100 gDW), ??-criptoxanthin (0.28 to 1.06 mg/100 gDW), ??-carotene (0.23 to 0.50 mg/100 gDW), and vitamin C (25.07 to 58.59 mg/100 gDW). These results indicate that stage of ripeness significantly influences the contents of bioactive compounds in papaya fruit.     

Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions

Melissa officinalis L. (lemon balm) is normally consumed as an infusion and presents therapeutic properties, such as sedative, carminative and antispasmodic, also being included in some pharmaceutical preparations. The phenolic profiles of different samples of lemon balm, prepared as infusions, were evaluated by HPLC-DAD-ESI/MS. The profiles were compared in order to understand the differences between cultivated, in vitro cultured and commercial (bags and granulated) samples. All the samples showed a similar phenolic profile, presenting differences only in the quantities found of each compound. Rosmarinic acid was the most abundant compound, being higher in commercial samples, especially in tea bag sample (55.68 mg/g of infusion) and lower in in vitro cultured sample (15.46 mg/g). Moreover, dimers, trimers and tetramers of caffeic acid were identified and quantified for the first time in lemon balm. Only one flavonoid, luteolin-3′-O-glucuronide was found in all the samples, ranging from 8.43 mg/g in commercial granulate sample to 1.22 mg/g in in vitro cultured sample. Overall, cultivated and in vitro cultured samples presented the lowest amounts of phenolic compounds (59.59 and 30.21 mg/g, respectively); otherwise, commercial samples showed the highest contents (109.24 mg/g for tea bag and 101.03 mg/g for granulate sample). The present study shows that infusion of lemon balm can be a source of phenolic compounds, known for their bioactive effects.     

Phenolic compounds in strawberry (Fragaria x ananassa Duch.) fruits: Composition in 27 cultivars and changes during ripening

Phenolic compounds in fruits of 27 cultivars of strawberry (Fragaria x ananassa Duch.) grown in Norway were characterised and quantified by HPLC-DAD-MSⁿ. Total phenolic content, calculated as the sum of the individual compounds, varied 2.3-fold among cultivars, i.e., from 57 to 133 mg/100 g of fw. There were significant differences among cultivars in concentration of all phenolic compounds. The highest variation between cultivars was found for cinnamoyl glucose (0.6-24.9 mg/100 g of fw). Concentration of anthocyanins, the most abundant class of phenolic compounds in the majority of the cultivars, varied from 8.5 to 65.9 mg/100 g of fw. Flavan-3-ols (11-45 mg/100 g of fw) and ellagitannins (7.7-18.2 mg/100 g of fw) contributed on average 28% and 14% to total phenolic contents in the strawberry cultivars, respectively. In three cultivars harvested at three stages of ripeness, anthocyanins and cinnamic acid conjugates were the compounds most affected by ripening. The anthocyanin profile for the individual cultivars was only slightly affected by ripening and growing conditions.     

Antioxidant, antiacetylcholinesterase and antimicrobial activities of Cymbopogon schoenanthus L. Spreng (lemon grass) from Tunisia

Aqueous extract, proanthocyanidin rich extract, and organic extracts of Cymbopogon schoenanthus L. Spreng (lemon grass) shoots from three different locations in South Tunisia were screened for their antioxidant, acetylcholinesterase and antimicrobial activities. In addition to the evaluation of these activities, the contents of flavonoids and total phenolic compounds were determined.Antioxidant activity measured by DPPH assay showed that the proanthocyanidin extract exhibited higher antioxidant activity than the aqueous extract. Extract concentration providing 50% inhibition (IC₅₀) ranged from 16.4 ± 6.8 μg/mL to 26.4 ± 6.8 μg/mL. The antioxidant activity was also determined using the β-carotene/linoleic acid bleaching test. The best results (IC₅₀ = 0.11 ± 0.10 mg/mL) were obtained with the proanthocyanidin extract of the plants collected from the desert region (Dhibat).The greatest acetylcholinesterase inhibitory activity (IC₅₀ = 0.23 ± 0.04 mg/mL) was exhibited by the ethyl acetate and methanol extracts of the plants collected from the mountainous region. It seems that extracts obtained with more polar solvents gave better results.The proanthocyanidin extracts showed a good antimicrobial activity against Streptococcus sobrinus at low concentration (MIC = 4 mg/mL). Therefore, these extracts could be used to prevent carious lesions by inhibiting S. sobrinus growth.     

Speciation of selenium in vitamin tablets using spectrofluorometry following cloud point extraction

Selenium is one of the trace and essential elements for good health but required only in a very narrow range. Hence, determination of selenium in trace level is important. In this work, cloud point extraction (CPE) with non-ionic surfactant Triton X-114 and the fluorometric ligand, 2,3-diaminonaphtalene (DAN) were used for extraction of trace amounts of organic and inorganic selenium species prior to their determination by spectrofluorometry. CPE parameters affecting complexation and phase separation were optimised. The limit of detection calculated by using nine replicate measurements of 0.020 mg/L Se solution after complexing with DAN and 10-fold CPE preconcentration was 2.1 μg/L. Accuracy of the method was checked using EnviroMat Waste Water, EU-L-2 as CRM and result was found to be in good agreement with the certified value. The suggested method can be used for selenium species of selenite, selenate, and total organic selenium at μg/L level.     

Flavonoids and other phenolic compounds in Andean indigenous grains: Quinoa (Chenopodium quinoa), kañiwa (Chenopodium pallidicaule) and kiwicha (Amaranthus caudatus)

The amount of phenolic acids, flavonoids and betalains in Andean indigenous grains, quinoa (Chenopodium quinoa), kañiwa (Chenopodium pallidicaule) and kiwicha (Amaranthus caudatus), was determined. The total amount of phenolic acids varied from 16.8 to 59.7 mg/100 g and the proportion of soluble phenolic acids varied from 7% to 61%. The phenolic acid content in Andean crops was low compared with common cereals like wheat and rye, but was similar to levels found in oat, barley, corn and rice. The flavonoid content of quinoa and kañiwa was exceptionally high, varying from 36.2 to 144.3 mg/100 g. Kiwicha did not contain quantifiable amounts of these compounds. Only one variety of kiwicha contained low amounts of betalains. These compounds were not detected in kañiwa or quinoa. Our study demonstrates that Andean indigenous crops have excellent potential as sources of health-promoting bioactive compounds such as flavonoids.     

Chemical characterization and antioxidant capacity of berries from Clidemia rubra (Aubl.) Mart. (Melastomataceae)

The flora of Latin America attracts gaining interest as it provides a plethora of still unexplored or under-utilized fruits that can contribute to human well-being due to their nutritional value and their content of bioactive compounds. Clidemia rubra (Aubl.) Mart. is a shrub belonging to the family of the Melastomataceae that grows preferably in a tropical climate. This paper comprises a nutritional characterization of the berries from Clidemia rubra and provides data on the phenolic compounds as well as the antioxidant capacity of the fruit. Findings in macronutrients like protein, carbohydrates, and fat were comparable to that of common berry fruits. Clidemia rubra berries seemed to be a good source for dietary fibers and some minerals (Ca, Mn, and Zn). In contrast, contents of titratable acids and ascorbic acid were low. The polyphenolic profile was determined by using HPLC-MS/MS in comparison to standard compounds. Noteworthy amounts of cyanidin 3-O-rutinoside (39.43 ± 1.66 mg/100 g fresh weight (FW)), delphinidin 3-O-rutinoside (23.74 ± 1.18 mg/100 g FW), cyanidin 3-O-glucoside (11.68 ± 0.56 mg/100 g FW), and delphinidin 3-O-glucoside (6.08 ± 0.35 mg/100 g FW) were found. Non-anthocyanin phenolic constituents were phenolic acids (gallic, protocatechuic, p-hydroxy-benzoic, vanillic, and caffeic acid), flavan-3-ols (epigallocatechin, epigallocatechin gallate, and epicatechin gallate), and 11 different myricetin and quercetin derivatives of which quercetin 3-O-arabinoside (5.26 ± 0.16 mg/100 g FW) and quercetin 3-O-rhamnoside (5.06 ± 0.08 mg/100 g FW) were dominating. Anthocyanins and ascorbic acid were mainly responsible for the antioxidant capacity of Clidemia rubra berries assessed with the total oxidant scavenging capacity (TOSC) assay.     

Comparative antioxidant activity of extracts from leaves,bark and catkins of Salix aegyptiaca sp.

Leaves, bark and catkins of Salix aegyptiaca L. were extracted into solvents of increasing polarity from cyclohexane (non-polar), butanol, ethanol and water (polar) and analysed for their antioxidant capacity, total phenol and flavonoids. The highest antioxidant activity (19 μg/ml IC₅₀ for inhibition of DPPH radical activity), total phenolic content (212 mg gallic acid equivalents/g of dried extract) and total flavonoid (479 mg catechin equivalents/g of dried extract) was observed in the ethanolic extract of bark. HPLC identification of phenolic compounds from the extracts indicated the presence of gallic acid, caffeic acid, vanillin and p-coumaric acid, myricetin, catechin, epigallocatechin gallate, rutin, quercetin as well as salicin. Our data indicates the presence of high amounts of phenols and flavonoids in different parts of S. aegyptiaca species and propose that extracts from this plant may be utilised as a source of health promoting antioxidants.     

Optimization of phenolics and dietary fibre extraction from date seeds

This work was conducted to optimise extraction conditions of phenolics and dietary fibre from date seeds. The effects of solvent to sample ratio, temperature, extraction time, number of extractions and solvent type on phenolic extraction efficiency were studied. At two-stage extraction, each stage 1 h duration at 45 °C with a solvent to sample ratio of 60:1, is considered optimum. Acetone (50%), and butanone were the most efficient solvents for extraction and purification, increasing the yield and phenolic contents of seed concentrate to 18.10 and 36.26 g/100 g, respectively. The total dietary fibre of seeds (57.87 g/100 g) increased after water and acetone extractions to 83.50 and 82.17 g/100 g, respectively. Nine phenolic acids (free and liberated) were detected in seeds with p-hydroxybenzoic (9.89 mg/100 g), protocatechuic (8.84 mg/100 g), and m-coumaric (8.42 mg/100 g) acids found to be among the highest. After extraction and purification, total phenolic acid content increased significantly from 48.64 to 193.83 mg/100 g. Protocatechuic, caffeic and ferulic acids were the major phenolic acids found in the concentrates. Based on this study, we believe date seed concentrates could potentially be an inexpensive source of natural dietary fibre and antioxidants and possibly used as a functional food ingredient.     

Chemical composition,antimicrobial potential against cariogenic bacteria and cytotoxic activity of Tunisian Nigella sativa essential oil and thymoquinone

We investigate in this work the chemical composition by GC–EIMS, the antibacterial and the cytotoxic activities of Tunisian Nigella sativa essential oil and its bioactive compound, thymoquinone, were tested against various clinical cariogenic bacteria (n = 30). Eighty-four compounds were identified in the essential oil. The major one was p-cymene (49.48%) whereas thymoquinone represented only 0.79%. The essential oil (2.43 mg/disc) containing only 3.35 μg of thymoquinone showed pronounced dose dependant antibacterial activity against Streptococcus mitis, Streptococcus mutans, Streptococcus constellatus and Gemella haemolysans (15.5 ± 0.707 mm). However, pure thymoquinone compound (150 μg/disk) was active against all the studied strains especially S. mutans and S. mitis (24.5 ± 0.71 and 22 ± 1.41 mm inhibition zones, respectively).     

Determination of steroidal saponins in different organs of yam (Dioscorea pseudojaponica Yamamoto)

Yams (Dioscorea spp.) are perennial trailing rhizome plants. Steroidal saponins, furostanol and spirostanol glycosides are the marked functional compounds in yams. In this investigation, a C18 solid phase extraction method was developed for yam saponins purification. The contents of saponins in various organs of yam (Dioscorea pseudojaponica Yamamoto) were also determined. Results showed that the recoveries of yam saponins extracted by the developed method were about 99.48–100.08% when the saponins (each saponin weighed 0.20, 0.50 and 1.00 mg) passing through the C18 cartridge. The extractive method could efficiently reduce the interferences from impurities in yam saponin extracts prior to HPLC analysis. The recoveries of added saponins in different yam organs were 98.34–99.92% for tuber flesh, 95.98–98.89% for tuber cortex, 97.89–99.44% for rhizophor, 93.82–98.01% for leaf and 93.87–97.65% for vine, respectively. The yam tuber cortex had the highest amount of saponins (582.53 μg/g dw), which was higher than that existed in the tuber flesh (227.86 μg/g dw) about 2.55 times. The contents of saponins in the rhizophor, leaf and vine of yam were 29.39, 24.41 and 23.96 μg/g dw, respectively.     

Antioxidant properties of chlorophyll-enriched and chlorophyll-depleted polyphenolic fractions from leaves of Vernonia amygdalina and Gongronema latifolium

This study investigated the optimum polyphenol extraction conditions for two Nigerian green leafy vegetables, Vernonia amygdalina (VA) and Gongronema latifolium (GL) that are widely used for food and medicinal purposes. Seven different solvents were used to extract the total phenolic contents of the leaves; 100% acetone, 80% acetone, 50% acetone, 100% ethanol, 70% ethanol, 100% methanol and 70% methanol and their total phenolic contents were determined. The results showed that the 80% acetone was significantly (p < 0.05) the most effective for polyphenol extraction from the leaves and generally the acetone-containing solvents were more effective than alcohol-containing solvents. The 80% acetone extract was fractionated on a silicic acid-packed column to give two main fractions: acetone eluted chlorophyll-enriched (flow-through) and ethanol eluted chlorophyll-depleted (column-bound). The ethanol fractions of VA and GL were significantly better (p < 0.05) DPPH radical scavengers than the acetone fractions. In contrast, the acetone fraction was a significantly better (p < 0.05) scavenger of hydroxyl radicals when compared to the ethanol fraction. Both acetone and ethanol fractions significantly (p < 0.05) and dose-dependently inhibited linoleic acid oxidation, though duration of inhibition was less at lower concentrations (10 and 25 ??g/ml) than at the highest concentration (50 ??g/ml).     

Extraction,distribution and characterisation of phenolic compounds and oil in grapeseeds

Five factors, extraction means, extraction repeats, extraction solvents, sample granularities, and grinding manners, were investigated and compared to optimise the extraction methods of phenolic compounds from grapeseed. Twenty minutes sequential sonication for three repeats was a preferred extraction means for assessment of phenolic content in grapeseed over three repeats 100-min sequential mechanical agitation. Methanol/water/HCl (70:29:1, v/v/v) was the best among the solvent compositions tested. Grinding granularity showed a significant (p = 0.05) effect on the yield of phenolic compounds when the sample was ground with a grinder and separated to different granularities by sieves. Grinding sample with mortar in liquid nitrogen could provide samples with uniform size and thus give a more consistent evaluation of phenolic compounds in grapeseed. Significant correlation coefficients (p < 0.01) among total phenols, total flavan-3-ols, and total proanthocyanidins contents in grapeseed extracts were observed. In addition, phenolic compounds and oil were very unevenly distributed in grapeseed as about 90% of phenolic compounds were observed in seed coat while about 60% of oil was in the endosperm and embryo.     

Syzygium aqueum leaf extract and its bioactive compounds enhances pre-adipocyte differentiation and 2-NBDG uptake in 3T3-L1 cells

The insulin-like and/or insulin-sensitising effects of Syzygium aqueum leaf extract and its six bioactive compounds; 4-hydroxybenzaldehyde, myricetin-3-O-rhamnoside, europetin-3-O-rhamnoside, phloretin, myrigalone-G and myrigalone-B were investigated in 3T3-L1 adipocytes. We observed that, S. aqueum leaf extract (0.04–5 μg/ml) and its six bioactive compounds (0.08–10 μM) at non-cytotoxic concentrations were effectively enhance adipogenesis, stimulate glucose uptake and increase adiponectin secretion in 3T3-L1 adipocytes. Clearly, the compounds myricetin-3-O-rhamnoside and europetin-3-O-rhamnoside showed insulin-like and insulin-sensitising effects on adipocytes from a concentration of 0.08 μM. These compounds were far better than rosiglitazone and the other isolated compounds in enhancing adipogenesis, stimulating 2-NBDG uptake and increasing adiponectin secretion at all the concentrations tested. These suggest the antidiabetic potential of S. aqueum leaf extract and its six bioactive compounds. However, further molecular interaction studies to explain the mechanisms of action are highly warranted.