Phytochemicals characterization of solvent extracts from taro-scented japonica rice bran

The major phytochemicals and antioxidant properties of taro-scented rice bran (TaiNung 71; TN71) extracts using 3 different solvents are characterized. Some progress is realized in creating an economic value for rice bran that has long been considered an agricultural waste. Various solvent extracts reveal the presence of phenolic compounds, oryzanols, tocopherols, and tocotrienols. Ethyl acetate (EtOAc) can extract more oryzanols (1.55 ± 0.20 g/kg rice bran). Meanwhile, the methanol (MeOH) extract possesses a higher yield in total contents (15.42 ± 1.41 g/kg bran), which includes phenolic compounds (2.69 ± 0.29 g gallic acid equivalent/kg bran), tocopherols (251 ± 26 mg/kg bran) and tocotrienols (111 ± 4 mg/kg bran). The MeOH extract exhibits more effective antioxidant activity against various oxidative systems in vitro, including the inhibition of linoleic acid peroxidation (33.89%), scavenging of DPPH radicals (83.88%), and reducing power. It is found that the yield, total content in phenolic compounds and tocols of the extracts increase with increasing Synder's polarity value and viscosity, which can then be used as the indices in isolation of the desired rice bran phytochemicals extracts.     

Antioxidant efficacy of phytochemical extracts from defatted rice bran in the bulk oil system

The antioxidant compounds oryzanols, tocols and ferulic acid were identified in the methanolic extracts of defatted rice bran (DRB) by high pressure liquid chromatography (HPLC). The crude methanolic extract (CME) was partially purified by re-extraction with acetone to give an acetone extract (AE). For further purification of the acetone extract, sequential solvent extraction was employed yielding a lipophilic phase (AE-LP) with hexane and a polar phase (AE-PP) with acetone. The antioxidant potential of the DRB extracts and their phytochemical constituents in bulk oils were evaluated using the Schall oven test (SOT) and differential scanning calorimetry (DSC). The extracts were effective in inhibiting lipid oxidation as assessed by peroxide value, diene value and p-anisidine value. The activity of the extracts with respect to the inhibition of primary oxidation products followed the order AE-PP > AE-LP = AE > CME with the activity of AE-PP being equivalent to that of butylated hydroxytoluene (BHT) at a 200 ppm level. However, tertiarybutylhydroquinone (TBHQ) was most active as compared to extracts and pure compounds with AE-PP showing about 45% of the activity of TBHQ at 200 ppm level. Defatted rice bran extracts proved to be effective even at the high temperature employed in DSC. The antioxidant efficacy of AE-PP was close to that of TBHQ and far greater than that of BHT at a 200 ppm level as evident from DSC results. The increase in activity with purification might be due to the enhanced levels of antioxidants in purified extracts compared to CME.     

Radical scavenging capacities and inhibition of human prostate (LNCaP) cell proliferation by Fortunella margarita

Lyophilised nagami kumquat (Fortunella margarita) powder was extracted with five different solvents. Dried extracts of EtOAc and MeOH-water (4:1, v/v) had the highest and lowest total phenolics, respectively, by Folin-Ciocalteu method. LC-MS analysis confirmed the presence of different levels of rutin, narirutin, poncirin, apigenin 8-C-rutinoside and 3′,5′,di-C-β-glucopyranosyl phloretin in all the extracts except n-hexane. EtOAc and MeOH extracts exhibited the highest and the lowest 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, respectively. The order of antioxidant capacity was found to be MeOH-water (4:1, v/v) > EtOAc > MeOH > n-hexane > acetone by phosphomolybdenum complex and oxygen radical absorbance capacity (ORAC) values. Among five extracts, n-hexane extract exhibited the highest inhibition of human prostate cancer (LNCaP) cells (86.4%) after 96 h at 100 μg/mL, followed by EtOAc (82.8%), MeOH (76.7%) and MeOH-water (4:1, v/v) (68.2%). Fragmentation of DNA suggests the ability of extracts to induce apoptosis in LNCaP cells. The cleavage of caspase-3 was the highest in n-hexane and EtOAc extracts, whereas the ratio of Bax/Bcl2 was the highest in MeOH and MeOH-water (4:1, v/v) extracts. The results of the present study were also supported by fluorescent images of LNCaP cells treated with kumquat extracts. The maximum cell proliferation inhibition activity of n-hexane extract may be due to the presence of one or cumulative effect of β-carotene, β-cubebene and hexadecanoic acid. Remaining four extracts exhibited differential antioxidant activity and cytotoxicity which may be due to the presence of various levels of rutin, narirutin, poncirin, apigenin-8-C-rutinoside and 3′,4′-di-C-β-glucopyranosyl phloretin in each extract.     

Nutritional and nutraceutical comparison of Jamaican Psidium cattleianum (strawberry guava) and Psidium guajava (common guava) fruits

Psidium cattleianum (strawberry guava) is one of many underutilised edible fruits that grow wild in Jamaica, and could potentially be commercially exploited to yield health and economic benefits. In this study, the total phenolics, proximate contents, and antioxidant, anti-inflammatory, and antimicrobial activities of P. cattleianum and P. guajava (common guava), a well-known species, were compared. Strawberry guavas were found to be superior to common guavas in antioxidant and antimicrobial activities, total phenolics and vitamin C content. They also possessed relatively high fibre content (24.9%). The hexane and ethyl acetate extracts of strawberry guavas showed cyclooxygenase-2 enzyme inhibitory activities of 18.3% and 26.5%, respectively (250 μg/mL), indicating anti-inflammatory activity. The EtOAc and MeOH extracts of P. guajava showed 56.4% (COX-2) and 44.1% (COX-1) inhibitory activity, respectively. Additionally, nine compounds were isolated from strawberry guava fruits, some of which demonstrated anti-inflammatory activity. These results indicate that strawberry guavas are beneficial for health.     

Dioscin: A synergistic tyrosinase inhibitor from the roots of Smilax china

In our study, we investigated the inhibition of mushroom tyrosinase in Smilax china. A methanol (MeOH) extract of S. china was partitioned into hexane, ethyl acetate (EtOAc) and water. Of the three fractions, EtOAc extract showed the strongest inhibition of tyrosinase activity with l-tyrosine or l-DOPA as a substrate. Two compounds were isolated from a final active fraction by activity-guided column chromatography. These compounds were identified as dioscin and oxyresveratrol by comparing their mass, ¹H- and ¹³C-NMR spectral data with those reported in the literature. Dioscin showed little inhibition activity of tyrosinase, whereas oxyresveratrol, a known tyrosinase inhibitor, showed a strong tyrosinase inhibitory activity. We discovered that a mixture of oxyresveratrol and dioscin (IC₅₀ = 5.1 and 5.7 μg/ml) highly increased the inhibition of tyrosinase activity with l-tyrosine or l-DOPA as the substrate as compared to either oxyresveratrol (IC₅₀ = 7.8 and 10.9 μg/ml) or dioscin (IC₅₀ > 100 and 100 μg/ml) alone.     

Microalgae of different phyla display antioxidant, metal chelating and acetylcholinesterase inhibitory activities

Methanol and hexane extracts from Tetraselmis chuii, Nannochloropsis oculata, Chlorella minutissima and Rhodomonas salina were evaluated for total phenolic contents, radical scavenging activity (RSA), metal chelating potential against copper and iron ions and acetylcholinesterase (AChE) inhibition. Only the methanol extracts contained phenolic compounds. The hexane extracts had the highest RSA. The extracts had a higher capacity to chelate Fe²⁺ ions, more pronounced in the lowest concentration of the hexane extracts with values ranging from 73.3 ± 3.3% (R. salina) to 97.5 ± 1.1% (N. oculata). The highest AChE inhibitory activity was found in the hexane extracts at 10 mg/ml of C. minutissima (79.3 ± 1.9%), T. chuii (85.7 ± 0.7%) and R. salina (81.5 ± 7.5%). GC-MS analysis indicated polyunsaturated fatty acids and steroids as the most abundant compounds in the hexane extracts. The species under study provide a valuable source of antioxidants, metal chelators and AChE inhibitors.     

Kinetic modeling of spice extraction from S. aromaticum and C. cassia

A mathematical model was developed for the extracts obtained from Syzygium aromaticum and Cinnamomum cassia with different particle size, solvent–solid ratios on extraction yield. Different particle sizes in the range of 2.8 mm to ?0.5 mm were employed and maximum extraction efficiency was achieved with particles of size ?0.5 mm. Among the solvent–solid ratios (20:1, 30:1, 40:1 and 50:1) ratio of 50:1 showed higher extraction yield. In the extraction kinetics, higher effective diffusivity value of 36.01 × 10⁻¹⁰ m²/s for S. aromaticum and 26.78 × 10⁻¹⁰ m²/s for C. cassia were achieved. Antioxidant values were determined and extracts prepared from ethanol showed higher scavenging activities for S. aromaticum and C. cassia as 78% and 85% respectively. Maximum phenolic content of 1.6 and 12.4 mg GAE/g of sample were achieved for S. aromaticum and C. cassia by hexane and water respectively.     

Comparison of the activities of hydrophilic anthocyanins and lipophilic tocols in black rice bran against lipid oxidation

The antioxidant capabilities of anthocyanin and tocol extracts from black rice bran were evaluated using an emulsion system containing either cholesterol (1.0 mg/ml) or fish oil (10 mg/ml). The cholesterol oxidation product, 7-ketocholesterol, increased to 180.1 μg/ml in the control emulsion after 168 h of oxidation, while it was only 15.4 and 39.0 μg/ml in the emulsions containing 1 μg/ml of the anthocyanin and tocol extracts, respectively; but below 1.2 μg/ml in the emulsion having 5 μg/ml of anthocyanins or tocols. In the fish oil emulsion, over 80% of C20:5 and C22:6 were oxidised after a 48 h incubation at 37 °C, while they were retained above 38% and 65% in the emulsions containing 10 μg/ml of anthocyanins and tocols, respectively, and above 85% in the emulsion containing 20 μg/ml of anthocyanins or tocols. Compared with the tocols extract, the capability of the anthocyanin extract was relatively greater in stabilising cholesterol but lower in inhibiting fatty acids oxidation.     

Chemical and economic evaluation of natural antioxidant extracts obtained by ultrasound-assisted and agitated bed extraction from jussara pulp (Euterpe edulis)

This work aimed to evaluate the influence of ultrasonic and agitated bed extractions on the chemical composition and manufacturing costs of extracts obtained from jussara (Euterpe edulis) pulp. The effects of extraction time (5–180 min), temperature (25–55 °C), ethanol concentration (0–90% in acidified water) and solvent/pulp ratio (5–30 mL/g) on the extraction yield, phenolic content, anthocyanin content, antioxidant capacity and manufacturing costs were assessed. The yields provided by the ultrasound-assisted and agitated bed extractions were not significantly different. The anthocyanins and phenolic compound yields were significantly affected by the extraction time, the ethanol concentration in water and the solvent/feed ratio, but not by the temperature. In general, the antioxidant capacity of the extracts displayed tendencies similar to the anthocyanin and phenolic compound yields. The production of crude extracts obtained by ultrasound and agitated bed extraction incurred greater manufacturing costs compared to the market prices of assai extracts.     

Characterisation of acid protease expressed from Aspergillus oryzae MTCC 5341

Aspergillus oryzae (MTCC 5341) has the largest expanse of hydrolytic genes, that includes 135 protease genes coding for alkaline, acid as well as neutral proteases. This study reports the purification and characterisation of an acid protease obtained from A. oryzae MTCC 5341. A. oryzae MTCC 5341 produces one of the highest reported acid protease activities reported so far (8.3 × 10⁵ U/g dry bran). The extracellular acid protease (47 kDa) was found to be active in the pH range 3.0–4.0 and stable in the pH range 2.5–6.5. Optimum temperature for activity was 55 °C. The protease was purified 17–fold with a yield of 29%. The enzyme was characterised to be an aspartate protease by inhibition studies, using pepstatin and its ability to activate trypsinogen. The enzyme cleaved the B-chain of insulin at L–V and Y–T residues.     

Validation of a method for simultaneous quantification of total carotenes and tocols in vegetable oils by HPLC

A normal-phase HPLC method for analysis of carotenes, tocopherols and tocotrienols has been developed and validated. In this work we presented a modification to the official AOCS method for analysis of tocols which allowed simultaneous quantification of the three groups of compounds, including carotenes. Analytes were separated using a gradient mobile phase (hexane and isopropanol) and with a gradient flow rate (1–2 mL min⁻¹). The column effluent was monitored by Photo Diode Array detector (PDA) set at 292 nm (tocols) and 455 nm (β-carotene) and by fluorescence detector set at an excitation wavelength of 290 and 330 nm emission. Inter- and intra-run accuracies and precision of the analytical method were better than ±15%. The lower limit of quantification was 5.0 mg L⁻¹ for the tocols and 0.1 mg L⁻¹ for carotenes. The method has been applied for the quantification of these compounds in Amazon oils.     

The analysis of phenolic constituents in glabrous canaryseed groats

Nineteen glabrous canaryseed samples, comprising brown- and yellow-coloured seeds, were investigated to determine the nature of phenolic constituents present. Total phenolic content (TPC) was determined, using the Folin–Ciocalteau assay. Flavonoid and phenolic acid compositions were determined, using high performance liquid chromatographic and mass spectrometric (LC–MS/MS) techniques. TPC ranged from 174 to 209 mg/100 g for canaryseed wholemeal samples. The canaryseed bran contained twice as much TPC as the wholemeal. The brown- and yellow-coloured whole canaryseeds exhibited the same flavonoid profiles. LC–MS/MS analysis showed that the canaryseed acetone extract was rich in flavonoid glycosides, with the bran being mainly composed of O-pentosyl isovitexin and the flour having a compound at m/z 468. No proanthocyanidins were detected in the 19 samples. Ferulic acid was the dominant phenolic acid, followed by caffeic and p-coumaric acids. The wholemeal obtained from the brown-coloured group had significantly higher contents of ferulic (>196 mg/kg) and caffeic (>96 mg/kg) acids in comparison to the yellow-coloured canaryseed group. The latter had ferulic and caffeic acids at levels less than 165 and 78 mg/kg, respectively, with one exception which had relatively higher levels (190 and 94 mg/kg). Whilst canaryseed flour contained significantly very low levels of ferulic acid (22–34 mg/kg), the bran was enriched in ferulic (593–766 mg/kg), caffeic (304–452 mg/kg) and p-coumaric (119–142 mg/kg) acids.     

Identification and analysis of isothiocyanates and new acylated anthocyanins in the juice of Raphanus sativus cv. Sango sprouts

The freeze-dried sprouts’ juice of Raphanus sativus (L.) cv. Sango was prepared and analysed for the first time. HPLC analysis of total isothiocyanates, after protein displacement, resulted in 77.8 ± 3.0 μmol/g of dry juice while GC–MS analysis of hexane and acetone extracts showed E- and Z-raphasatin (8.9 and 0.11 μmol/g, respectively) and sulforaphene (11.7 μmol/g), summing up to 20.7 ± 1.7 μmol/g of free isothiocyanates. Sprouts’ juice contained an unprecedented wealth of anthocyanins and a new fractionation methodology allowed us to isolate 34 mg/g of acylated anthocyanins (28.3 ± 1.9 μmol/g), belonging selectively to the cyanidin family. Analysis was performed by HPLC–PDA–ESI–MSⁿ and extended to deacylated anthocyanins and aglycones, obtained, respectively, by alkaline and acid hydrolysis. This study identified 70 anthocyanins, 19 of which have never been described before and 32 of which are reported here in R. sativus for the first time. Sango radish sprouts are exceptional dietary sources of heath-promoting micronutrients.     

Extraction of active ingredients from green tea (Camellia sinensis): Extraction efficiency of major catechins and caffeine

The effect of different extraction set-ups that influence the extraction efficiency of catechins and caffeine from green tea leaves (variety Fanning Belas, China) were studied using different aqueous and pure solvents (acetone, ethanol, methanol, acetonitrile, water), different temperatures (60, 80, 95 and 100 °C) and times (5–240 min). Raw extracts were analysed for contents of major catechins (EC, EGC, ECG, EGCG), caffeine, proanthocyanidins and flavonols (myricetin, caempherol, quercetin). Starting material was found to contain 191 g major catechins/kg material, 36 g caffeine/kg material and 5.2 g flavonols/kg material on a dry mass basis. The content of major catechins in green tea extracts varied from approximately 280–580 g/kg dry extract, with extraction efficiencies of major catechins varying from 61% to almost 100%. Content of caffeine in extract was in the range of 75 g/kg, where its extraction efficiency varied from 62% to 76%. Average extraction yield was 30% with exceptions when using pure acetone and acetonitrile, where extraction yield was about 3%. Contents of flavonols and proanthocyanidins were in the ranges 6–20 and 12–19 g/kg, respectively. Different extraction procedures with water were also investigated and optimal conditions determined: maximum achieved extraction efficiency of catechins with water was obtained at 80 °C after 20 min (97%) and at 95 °C after 10 min of extraction (90%). Degradation of catechins was observed at higher extraction temperatures and with prolonged extraction times. Using a lower ratio of solvent to material, extraction efficiencies were increased by applying a multi-step extraction procedure. Optimal extraction procedure was then performed using decaffeinated green tea leaves, which were obtained by high-pressure extraction with CO₂, when 98% of caffeine was selectively isolated without significant impact on valuable catechins.     

Principal phenolic phytochemicals and antioxidant activities of three Chinese medicinal plants

The principal antioxidant components and content of cinnamon (Cinnamomum cassia), turmeric (Curcuma longa) and golden thread (Coptidis rhizoma) extracts were determined using high performance liquid chromatography (HPLC) with UV detection. In general, C. cassia, C. longa and C. rhizoma extracts from domestic Taiwan were rich in cinnamaldehyde, curcumin, and berberin, respectively. The contents of cinnamaldehyde, curcumin, and berberin in the acetone extracts were 1911, 2029, and 840 mg l⁻¹, respectively. The Folin–Ciocalteu method was used to measure the total phenolic concentrations of extracts, which had the content of 9.6 (C. cassia), 2.6 (C. longa), and 4.3 (C. rhizoma) mM l⁻¹. In addition, DPPH radical-scavenging, ferric-reducing antioxidant power (FRAP), and ferric thiocyanate (FTC) assays were employed to measure antioxidant activities. The C. cassia fresh extracts had higher antioxidant activities which were 84–90% (DPPH), 17–33 μmol l⁻¹ (FRAP), and 53–82% (FTC). The activities of C. longa fresh extracts were 22–44% (DPPH), 7–11 μmol l⁻¹ (FRAP), and 53–81% (FTC) while C. rhizoma were 53–64% (DPPH), 18–26 μmol l⁻¹ (FRAP), and 59–82% (FTC).     

Silphium L. extracts – composition and protective effect on fatty acids content in sunflower oil subjected to heating and storage

The influence of ethanol and hexane extracts from leaves, inflorescences, and rhizomes of Silphium perfoliatum, Silphium trifoliatum, Silphium integrifolium on fatty acid content changes in sunflower oil subjected to heating and storage was studied in comparison to the synthetic antioxidant – butylated hydroxyanisole (BHA). A positive effect of extracts made of above-ground and underground organs of Silphium on the durable quantitative composition of fatty acids was proven. Tested extracts elevated the value of change inhibition with reference to linoleic acid to a level comparable with BHA, and sometimes, in appropriate systems, they were characterized by better values (for oil stored for 180 days at room temperature, the inhibition coefficient for linoleic acid changes reached 4.6% for 0.04% BHA, and 7.09% for hexane extract made of S. trifoliatum inflorescences, 400 μl/2 g; for oil heated for 120 h, the inhibition coefficient of linoleic acid changes amounted to 11.32% for 0.06% BHA, and 15.69% for hexane extract made of S. perfoliatum rhizomes, 600 μl/2 g). It was found that active substances groups such as phenolic acids, flavonoids and terpenes were present in tested extracts.     

Content of antioxidant hydroquinones substituted by β-1,6-linked oligosaccharides in wheat milled fractions,flours and breads

The objective of this study was to quantify seven antioxidant hydroquinones, substituted by β-1,6-linked oligosaccharides, in wheat-based food products by HPLC. These novel compounds were discovered and characterised in our previous study. All seven compounds were quantified in wheat germ and bran. The most abundant compound, 4-hydroxy-3-methoxyphenyl β-d-glucopyranosyl-(1 → 6)-β-d-glucopyranosyl-(1 → 6)-β-d-glucopyranoside, was present at 6.3 g/kg in germ and 0.7 g/kg in bran. In whole-wheat grains, as well as in several kinds of wheat flour and wheat bread, these antioxidants were present at much lower concentrations; only two of them could be quantified. None of these compounds were detected in low-protein flour mixture. Their total content in analysed wheat products decreased in the following order: germ > bran > whole grains ∼ whole grain flours > refined flours > refined bread ? low-protein flour mixture. It correlated well with the antioxidant capacity of water–ethanol extracts of these products determined by Trolox equivalent antioxidant capacity (TEAC) assay (r² = 0.984). The contribution of the analysed compounds to the total antioxidant capacity of wheat germ extract was 52%, whereas the contribution of the most abundant compound was 38%.     

Partial extraction method for the rapid analysis of total lipids and γ-oryzanol contents in rice bran

Total lipids and γ-oryzanol in rice bran were determined by a partial extraction method. The results agreed well with the conventional total extraction methods. The proposed method uses fewer hazardous organic solvents, takes a shorter extraction time and requires no special extraction apparatus. Total lipids and γ-oryzanol in nine rice bran varieties were analysed by the developed technique. Daw Dum 5647 had the highest total lipids and γ-oryzanol while the lowest content was found in KD XBT 313-19-1-1 and SP XBT 43-7, respectively. The adsorption coefficient (K_d) of the lipids and γ-oryzanol, between hexane and bran, at 30 °C are between 1.16 and 2.00 and 2.02 and 2.65, respectively (depending on the moisture content of the bran). From the K_d values, it was estimated that about 92–95% of the lipids and 95–96% of the γ-oryzanol were extracted into hexane at a 10:1 (v/w) ratio of hexane to bran. The effect of solvents on the extraction of γ-oryzanol from rice bran was also studied. It was found that isopropanol was the most suitable solvent for extraction and determination of γ-oryzanol in rice bran. It showed better agreement with the total extraction method.