Recovery of hesperidin and narirutin from waste Citrus unshiu peel using subcritical water extraction aided by pulsed electric field treatment

The objective of this study was to identify whether the efficacy of extracting hesperidin and narirutin from Citrus unshiu peel by-products can be increased by combining pulsed electric field (PEF) and subcritical water extraction (SWE). The samples were treated with a PEF at a strength of 3 kV/cm for 60 and 120 s. Subsequent SWE was conducted at extraction temperatures of 110–190 °C for 3–15 min. The concentration of hesperidin was highest at 46.96 ± 3.37 mg/g peel (dry basis) after PEF treatment at 120 s, combined with SWE at 150 °C for 15 min, while that of narirutin peaked at 8.76 ± 0.83 mg/g after PEF treatment at 120 s, integrated with SWE at 190 °C for 5 min. The concentrations of both hesperidin and narirutin increased with PEF treatment time. The PEF increased the amounts of hesperidin and narirutin extracted by 22.1% and 33.6%, respectively. This study demonstrate the potential of PEF pretreatment for enhancing the SWE of flavonoids from C. unshiu peel.     

Quantification of main phenolic compounds in sweet and bitter orange peel using CE–MS/MS

The food and agricultural products processing industries generate substantial quantities of phenolics-rich subproducts, which could be valuable natural sources of polyphenols. In oranges, the peel represents roughly 30% of the fruit mass and the highest concentrations of flavonoids in citrus fruit occur in peel. In this work we have carried out the characterisation and quantification of citrus flavonoids in methanolic extracts of bitter and sweet orange peels using CE–ESI–IT–MS. Naringin (m/z 579.2) and neohesperidin (m/z 609.2) are the major polyphenols in bitter orange peels and narirutin (m/z 579.2) and hesperidin (m/z 609.2) in sweet orange peels. The proposed method allowed the unmistakable identification, using MS/MS experiments, and also the quantification of naringin (5.1 ± 0.4 mg/g), neohesperidin (7.9 ± 0.8 mg/g), narirutin (26.9 ± 2.1 mg/g) and hesperidin (35.2 ± 3.6 mg/g) in bitter and sweet orange peels. CE coupled to MS detection can provides structure-selective information about the analytes. In this work we have developed a CE–ESI–IT–MS method for the analysis and quantification of main phenolic compounds in orange peels.     

Isolation of hesperidin from peels of thinned Citrus unshiu fruits by microwave-assisted extraction

Simultaneous extraction by microwave-irradiation and crystallisation were performed in the same pot of solvent of 70% (v/v) aqueous ethanol for isolation of hesperidin from thinned immature fruit peels of Citrus unshiu as refining of Citrus waste biomass. The hesperidin content in immature fruits peels was about 3.2-fold higher than that of mature fruit. After microwave-assisted extraction (MAE), the yield of hesperidin reached 58.6 mg/g, which was comparable to the amount obtained after extraction using DMSO:methanol (1:1, v/v) as a solvent for 30 min at room temperature. Heating temperature and time for isolation of hesperidin crystallites were optimised as 140 °C and 8 min by using response surface methodology. Under this optimal condition, 86.8% (47.7 mg/g) of total hesperidin was isolable by MAE and low-temperature storage (5 °C, 24 h).     

Antioxidative and anti-inflammatory properties of Citrus sulcata extracts

Citrus sulcata was subjected to ultrasound, high-pressure, and Soxhlet extractions. The antioxidant and anti-inflammatory activities of the extracts were evaluated qualitatively and quantitatively. The antioxidant content of the peel extract was twice that of the fruit extract. The quantitative analysis showed that the narirutin and hesperidin contents in the peel extracts were 8.8 and 7.5 mg/100 g, respectively. These extracts had a total phenolic content of 112.22 ± 2.89 gallic acid equivalent (GAE) mg/100 g, a total flavonoid content of 54.09 ± 1.01 rutin equivalent (RE) mg/100 g, DPPH radical scavenging activity of 46%, and antioxidant activity of 213.25 ± 2.82 μM of Trolox equivalents (TEAC). C. sulcata extracts could prevent hydrogen peroxide (H₂O₂)-induced oxidative stress, reduce expression of the inflammatory markers nuclear Factor kappa B (NFκB) and phosphorylated IκBα (p-IκBα) in A549 human lung carcinoma cells, and inhibit Lipopolysaccharide (LPS)-induced THP-1 monocyte differentiation to an extent of 85%.     

Antimicrobial activity of acid-hydrolyzed Citrus unshiu peel extract in milk

Citrus fruit (Citrus unshiu) peels were extracted with hot water and then acid-hydrolyzed using hydrochloric acid. Antimicrobial activities of acid-hydrolyzed Citrus unshiu peel extract were evaluated against pathogenic bacteria, including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes. Antilisterial effect was also determined by adding extracts at 1, 2, and 4% to whole, low-fat, and skim milk. The cell numbers of B. cereus, Staph. aureus, and L. monocytogenes cultures treated with acid-hydrolyzed extract for 12 h at 35°C were reduced from about 8 log cfu/mL to <1 log cfu/mL. Bacillus cereus was more sensitive to acid-hydrolyzed Citrus unshiu peel extract than were the other bacteria. The addition of 4% acid-hydrolyzed Citrus unshiu extracts to all types of milk inhibited the growth of L. monocytogenes within 1 d of storage at 4°C. The results indicated that Citrus unshiu peel extracts, after acid hydrolysis, effectively inhibited the growth of pathogenic bacteria. These findings indicate that acid hydrolysis of Citrus unshiu peel facilitates its use as a natural antimicrobial agent for food products.     

Determination of the change of flavonoid components as the defence materials of Citrus unshiu Marc. fruit peel against Penicillium digitatum by liquid chromatography coupled with tandem mass spectrometry

A healthy fruit peel of Citrus unshiu Marc. and one infected by Penicillium digitatum were analysed for flavonoids via high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC–MS/MS) in the positive mode with selected ion monitoring (SIM). Among 16 flavonoid components characterised in C. unshiu Marc., four flavanones and nine flavones were identified for the first time. The identified compounds were quantified by HPLC–UV. To investigate the function of the flavonoids as defence materials, the flavonoid content change of the fruit peel inoculated with P. digitatum was monitored by HPLC. The flavonoid concentration in the infected fruit peel decreased initially after the infection and then gradually increased before finally progressively decreasing.     

Bioactive compounds and antioxidant capacities in different edible tissues of citrus fruit of four species

Different edible tissues of citrus fruit, namely juice sacs (JS), segment membrane (SM), and segment (Seg), of four species, were examined for contents of bioactive compounds and total antioxidant capacities (TAC) by ferric reducing antioxidant power (FRAP) assay. Two flavanones (naringin and hesperidin) were identified by HPLC; hesperidin accounted for 18.5–38.5% of the total phenolics in the species Citrus unshiu, Citrus reticulata, and Citrus sinensis, while naringin was only found in Citrus changshanensis and it accounted for 53.7% of the total phenolics in SM of this species. In SM of all selected species, the contents of phenolic compounds and TAC were significantly higher than those in JS and Seg. Highest total phenolics, total flavonoids, naringin, and TAC were found in SM of C. changshanensis, while the highest carotenoid content was found in JS of C. reticulata. The contribution of vitamin C to TAC ranged from 26.9% to 45.9% in JS and Seg of all selected species. In SM, however, a high contribution from hesperidin was observed in C. unshiu (54.0%), C. sinensis (46.7%) and C. reticulata (30.0%). The results indicated that SM of citrus fruit were high in contents of bioactive compounds and TAC; it is thus recommended to consume citrus fruit with all edible tissues rather than juice or JS alone.     

Effect of air-drying temperature on physico-chemical properties of dietary fibre and antioxidant capacity of orange (Citrus aurantium v. Canoneta) by-products

Dehydration promoted important modifications affecting both the physico-chemical properties of dietary fibre (DF) and the antioxidant capacity of orange by-products (peel and pulp remaining after juice extraction). The significance of such changes was largely dependent on the air-drying temperature used (from 30 °C to 90 °C). The major modifications on the DF components were observed when either extended drying periods, i.e. at lower temperatures, or elevated drying temperatures were applied. Dehydration around 50–60 °C apparently promoted the minor disruption of cell wall polymers, in particular of pectic substances. Pulp samples exhibited higher values of swelling (SW) and fat adsorption capacity (FAC) than those derived from orange peel. Although, significant decreases in water retention capacity (WRC), FAC and solubility values were detected for both by-products as the air-drying temperature increased. The antioxidant capacity associated to dehydrated citrus by-products was significantly higher for orange peel than for pulp samples. In general, the by-products studied proved to be quite resistant to the different heat treatments applied within the range of 40–70 °C. In overall, the study shows that, in order to preserve either the DF quality and/or the antioxidant capacity, air-drying temperature should be controlled since both types of compounds, DF components and antioxidants, might be degraded or modified either when extended drying periods and/or high drying temperatures are applied.     

Optimisation of gelatine extraction from hoki (Macruronus novaezelandiae) skins and measurement of gel strength and SDS–PAGE

The extraction of hoki (Macruronus novaezelandiae) gelatine was optimised using Response Surface Methodology (RSM) and gel strength and SDS–PAGE were evaluated. The optimum conditions for extraction were 0.75 M NaCl for 9 min of pre-treatment time and hot water extraction at 49.3 °C for 60 min. Results showed that the predicted yield by RSM (17.4%) closely matched the experimental yield of 17.6%. SDS–PAGE showed that hoki gelatine contained higher molecular weight subunits (∼191 kDa) but lower gel strength (197 ± 5 g) than those from porcine (307 ± 8.4 g) or bovine (273 ± 16.1 g) gelatine determined at 7 °C. However, hoki gelatine gel strength was significantly higher than those from other cold-water fish species reported in literature, which could account for the slight differences in methodologies reported.     

Subcritical water extraction of flavonol quercetin from onion skin

Subcritical water could be an excellent alternative to organic solvent as a medium for extracting flavonol quercetin, due to its temperature-dependent selectivity, safety, efficiency of recovery, and lower cost. This study investigated the application of subcritical water extraction (SWE) of quercetin from onion skin and evaluated the effect of key operation conditions by varying the temperature (100-190 °C), extraction time (5-30 min), and mixture ratio of onion skin and diatomaceous earth (DE) (0.5:3.5-2:2) under high pressure (90-131 bar). The maximum yield of quercetin (16.29 ± 0.75 mg/g onion skin) was obtained at extraction temperature of 165 °C, extraction time of 15 min, mixture ratio of 1.5:2.5 for onion skin and DE. The SWE was compared with three conventional extraction methods in terms of the efficiency. The quercetin yield by SWE was over eight-, six-, and fourfold greater than those obtained using the ethanol, methanol, and water-at-boiling-point extraction methods, respectively.     

Enzymatic water extraction of taxifolin from wood sawdust of Larix gmelini (Rupr.) Rupr. and evaluation of its antioxidant activity

An enzyme incubation–water extraction (EI–WE) method was developed and optimised for the extraction of the natural antioxidant taxifolin and of the total flavonoids from wood sawdust of Larixgmelini (Rupr.) Rupr. A factorial design and a central composite design approach were used for method optimisation. Optimal conditions were 0.5 mg/ml cellulase and 0.5 mg/ml pectinase, a pH of 5.0, a temperature of 32 °C and 18 h incubation time. The flavonoids and taxifolin were extracted in hot water at 50 °C for 30 min, with a solid to liquid ratio of 1:20. Under optimised conditions, the yields of taxifolin and total flavonoids increased from 1.06 ± 0.08 to 1.35 ± 0.04 mg/g and 4.13 ± 0.17 to 4.96 ± 0.29 mg/g, respectively. DPPH and BHT assays revealed that the EI–WE samples had 1.8- and 1.68-fold higher antioxidant activities than the controls. SEM results revealed the structural disruption of wood sawdust with enzyme incubation.     

Ultrasound-assisted extraction of polyphenols (flavanone glycosides) from orange (Citrus sinensis L.) peel

The present study reports on the extraction of polyphenols especially flavanones from orange (Citrus sinensis L.) peel by using ethanol as a food grade solvent. After a preliminary study showing that the best yield of extraction was reached for a particle size of 2 cm², a response surface methodology (RSM) was launched to investigate the influence of process variables on the ultrasound-assisted extraction (UAE) followed by a central composite design (CCD) approach. The statistical analysis revealed that the optimised conditions were a temperature of 40 °C, a sonication power of 150 W and a 4:1 (v/v) ethanol:water ratio. The high total phenolic content (275.8 mg of gallic acid equivalent/100 g FW), flavanone concentrations (70.3 mg of naringin and 205.2 mg of hesperidin/100 g FW) and extraction yield (10.9 %) obtained from optimised UAE proved its efficiency when compared with the conventional method. Furthermore, the antioxidant activity determined by the DPPH and ORAC tests confirmed the suitability of UAE for the preparation of antioxidant-rich plant extracts.     

Analysis of ginsenosides in Panax ginseng in high pressure microwave-assisted extraction

High pressure microwave assisted extraction (HPMAE) was applied to extract the ginsenosides from Panax ginseng root. The influences of extraction solvent, extraction pressure and extraction time were individually investigated. HPMAE has been compared with other extraction methods, including Soxhlet extraction, ultrasound-assisted extraction and heat reflux extraction. The determination of ginsenosides was performed by HPLC–ESI-MS. The results indicated that the HPMAE not only took a shorter time but also afforded higher extraction yields of ginsenosides, especially ginsenoside Rb₁, Rc, Rb₂ and Rd. Furthermore, the neutral ginsenosides and malonyl ginsenosides in Panax ginseng root extracts by HPMAE were investigated. The malonyl ginsenoside m-Rb₁, m-Rc, m-Rb₂ and m-Rd degraded in HPMAE at 400 kPa (109–112 °C) in 70% (v/v) ethanol–water and at 600 kPa (112–115 °C) in methanol, and transformed into corresponding neutral ginsenoside Rb₁, Rc, Rb₂ and Rd. Using water as extraction solution, the neutral ginsenosides degraded under HPMAE at 400 kPa (135–140 °C), and transformed into less polarity rare ginsenosides.     

Ionizing radiation and marketing simulation on bioactive compounds and quality of grapefruit (Citrus paradisi c.v. Rio Red)

Bioactive compounds in citrus fruits have been shown to be protective against chronic diseases such as cancer and heart disease, but their levels may be affected by postharvest treatments such as storage and irradiation. In this study, grapefruits were exposed to gamma irradiation at 0, 150 and 300 Gy and then stored at 10 °C for 36 d, followed by an additional 20 d at 20 °C. Flavonoid content, terpenoid content, quality (acidity and total soluble solids) and phenylalanine ammonia-lyase (PAL) activity were evaluated at regular intervals during storage. Irradiation and storage affected (P ? 0.05) the levels of bioactive compounds in grapefruit; however, the effect of storage was prominent. Irradiation differentially affected the flavonoid content of pulp and peel. Fruits exposed to 300 Gy had higher (P ? 0.01) narirutin content in peel compared to control fruits at 12 and 56 d after storage. While storage increased the d-limonene and myrcene content in all treatments, control fruit had higher terpenoid content at the end of the storage. PAL activity was found to be in traces in the peel. In general, irradiation or storage had no considerable effect on total soluble solids; however, acidity decreased (P ? 0.05) with storage.     

Optimisation of the aqueous extraction conditions of phenols from meadowsweet (Filipendula ulmaria L.) for incorporation into beverages

Meadowsweet was extracted in water at a range of temperatures (60–100 °C), and the total phenols, tannins, quercetin, salicylic acid content and colour were analysed. The extraction of total phenols followed pseudo first-order kinetics, the rate constant (k) increased from 0.09 ± 0.02 min⁻¹ to 0.44 ± 0.09 min⁻¹, as the temperature increased from 60 to 100 °C. An increase in temperature from 60 to 100 °C increased the concentration of total phenols extracted from 39 ± 2 to 63 ± 3 mg g⁻¹ gallic acid equivalents, although it did not significantly affect the proportion of tannin and non-tannin fractions. The extraction of quercetin and salicyclic acid from meadowsweet also followed pseudo first-order kinetics, the rate constant of both compounds increasing with an increase in temperature up until 90 °C. Therefore, the aqueous extraction of meadowsweet at temperatures at or above 90 °C for 15 min yields extracts high in phenols, which may be added to beverages.     

Supercritical carbon dioxide extraction of polyunsaturated fatty acids from Northern shrimp (Pandalus borealis Kreyer) processing by-products

This study investigated the potential of Northern shrimp (Pandelus borealis Kreyer) by-products as a source of omega-3 polyunsaturated fatty acids (ω-3 PUFAs). The by-products (heads, shell and tail) of processing accounted for approximately 50–60% of the catch. Supercritical CO₂ extraction (SFE) of the by-products at 35 MPa and 40 °C generated a deep red oil, rich in ω-3 PUFAs, specifically 7.8 ± 0.06% eicosapentaenoic acid (EPA) and 8.0 ± 0.07 % docosahexaenoic acid (DHA).     

Simultaneous determination of flavanones,hydroxycinnamic acids and alkaloids in citrus fruits by HPLC-DAD–ESI/MS

A simple and accurate method has been developed to simultaneously separate and determine 10 bioactive compounds in citrus fruits by high-performance liquid chromatography coupled with diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD–ESI/MS). This HPLC assay was performed on a reversed-phase C18 column with acetonitrile and 0.1% (v/v) aqueous formic acid as mobile phase. DAD has been performed at 273, 283 and 324 nm for quantification of the alkaloids, flavanones and hydroxycinnamic acids. MS was also employed to identify the each analyte. Ten analytes (naringin, hesperidin, neohesperidin, ferulic acid, p-coumaric acid, chlorogenic acid, caffeic acid, octopamine, synephrine and tyramine) demonstrated good linearity (r ? 0.9990) in a relatively wide concentration range. The method revealed high average recovery (range, 92.1–97.9%) and good precision with interday and intraday variations with less than 4.71%. The limits of detection (LODs) ranged from 0.02 to 0.11 μg/ml, while the limits of quantification (LOQs) were defined in the range of 0.08–0.39 μg/ml. The proposed method has been successfully applied to analyse three types of bioactive constituents in eight citrus hybrids pulps and eight citrus hybrids peels, which has been successfully cultivated in China.     

Effect of extraction conditions on the quality characteristics of pectin from passion fruit peel (Passiflora edulis f. flavicarpa L.)

Passion fruit (Passiflora edulis f. flavicarpa L.) yellow variety is composed of 50-55 g peel per 100 g of fresh fruit which is discarded as waste during processing. Utilization of passion fruit peel for pectin extraction was studied. Passion fruit peel obtained after juice extraction was blanched in boiling water for 5 min, dehydrated in a cross flow hot air drier at 60 ± 1 °C to a moisture content of 4 g/100 g of dried peel. The dehydrated passion fruit peel was used for extraction experiments of pectin. The effect of pH, peel to extractant ratio, and number of extractions, extraction time and temperature on the yield and quality characteristics of pectin were investigated. The optimized conditions for extraction of pectin from passion fruit peel yielded 14.8 g/100 g of dried peel. Pectin extracted from the dried peels had a methoxyl content of 9.6 g/100 g, galacturonic acid content of 88.2 g/100 g and jelly grade of 200. Extraction of pectin from dried peels of passion fruit may be considered for effective utilization of passion fruit processing waste.     

The optimisation of extraction of antioxidants from potato peel by pressurised liquids

Response Surface Methodology was used to optimise the solid–liquid extraction and Pressurised Liquid Extraction of polyphenols from industrially generated potato peel. Efficiency of extraction was optimised by measuring antioxidant activity, phenol content and the level of caffeic acid. Conditions for optimal antioxidant activity as measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay were 75% ethanol, 80 °C and 22 min with solid–liquid extraction, resulting in an optimum activity of 352 mg Trolox Equivalents/100 g DW potato peel. In comparison, the use of Pressurised Liquid Extraction resulted in an optimum activity of 339 mg Trolox Equivalents/100 g DW potato peel at 70% ethanol and 125 °C. Therefore the use of Pressurised Liquid Extraction did not enhance extraction in comparison to solid–liquid extracts, but using aqueous ethanol as extraction solvent recovered a higher level of polyphenols than when using 100% methanol.