超声波辅助法提取石榴皮中总多酚工艺

以石榴皮为原料,采用正交试验对石榴皮多酚的提取工艺进行研究。确定利用超声波辅助法提取石榴皮中多酚类物质的最佳工艺条件。结果表明：料液比1:20(g/mL)、30℃条件下超声波(固定功率100W)作用20min、提取1次,总多酚提取率最高,最终测定石榴皮多酚含量为14.06mg/g。     

超声处理对石榴皮多酚提取效果的影响

石榴皮是一种易获得的、廉价的,但含有丰富多酚的废料。采用超声辅助提取法从石榴皮中提取多酚,优化工艺参数并利用高效液相色谱法(HPLC)测定石榴皮中多酚的成分和含量,同时研究了石榴皮表面在超声处理下的变化。研究结果表明,在超声条件下,用水作为溶剂有更高的多酚得率,比甲醇等有机溶剂提取效果更好。在优化条件,即液固比(mL∶g)为60∶1,超声功率为400 W和超声时间为25 min时,多酚的最佳得率为39.30 mg/g(以没食子酸为标准),高于传统溶剂提取法的得率(32.58 mg/g)。HPLC分析得到石榴皮多酚中安石榴苷含量最高,其次是石榴皮鞣素,并且超声处理能较大地强化提取效果。石榴皮表面显微观察证明,超声能够显著破坏石榴皮表面而影响植物组织完整性,使多酚更多地释放出来。     

Study of flavonoids in aqueous spinach extract using positive electrospray ionisation tandem quadrupole mass spectrometry

The presence of three flavonols (quercetin, kaempferol, myricetin) and two flavones (apigenin, luteolin) were investigated in the extract of fresh spinach leaves. Aqueous spinach extracts were prepared with a leaf/water ratio of 1:2 at 80 °C for 30 min stirring. Ferric ammonium sulphate method was used for measuring total polyphenols in the extracts and expressed as catechins and tannic acid equivalents. The flavonoids glycosides in the extract were hydrolysed to their aglycons with 1.2 M HCl in boiling 50% water methanol solution. The resulting aglycons were identified and quantified by a C18 reverse phase high-performance liquid chromatography (RP-HPLC). Furthermore, the results were confirmed by HPLC coupled to an electrospray ionisation tandem triple-quadrupole mass spectrometer ESI-MS performing low-energy collision induced dissociation (CID-MS/MS) in the collision cell (HPLC-ESI-MS/MS). Analyses were made in the multiple reaction monitory (MRM) mode. Results showed that total polyphenols contents in fresh spinach leaves were 270 mg kg⁻¹ and 390 mg kg⁻¹ as tannic acid and catechin equivalents, respectively, in which, major flavonoids aglycons were apigenin (170 mg kg⁻¹), quercetin (50 mg kg⁻¹) and kaempferol (30 mg kg⁻¹).     

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.     

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).     

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.     

Purification,antioxidant activity and protein-precipitating capacity of punicalin from pomegranate husk

Punicalin is a kind of ellagitannin, existing in pomegranate husk, and has shown remarkable biological activities. A rapid and large-scale separation method of punicalin from pomegranate husk was established, using medium pressure liquid chromatography (MPLC). The optimal mobile phase consisted of 5% methanol and 0.1% (v/v) TFA in water, and the optimal loading amount and flow rate were 1.0 g and 80 ml/min, respectively. Under this condition, 339 mg of 95.9% punicalin could be obtained in 40 min. 59.7 mg of 78.0% gallic acid could be separated simultaneously. This method was practical for industrial utilisation of pomegranate husk. Afterwards, the antioxidant and protein-precipitating capacities of the purified punicalin, together with punicalagin, were evaluated. Results showed that punicalin had strong antioxidant activity, and it exhibited a low affinity for protein. This suggested that the antioxidant of punicalin would not be greatly masked by tannin–protein precipitation in application, and hence confirmed punicalin to be a promising antioxidant.     

石榴皮提取物的抗氧化作用研究

分别以水、甲醇、丙酮和乙酸乙酯为溶剂,对石榴皮中的多酚进行了提取,得到4种提取物。采用油脂过氧化值(POV)法和二苯三硝基苯肼(DPPH)自由基法研究了提取物的抗氧化活性。结果显示,4种提取物均具有抗油脂过氧化和清除DPPH自由基的能力,其抗氧化活性随添加量的增加而增加。4种提取物中,丙酮提取物的抗油脂过氧化作用最大,其次为水提取物和甲醇提取物,乙酸乙酯提取物的抗油脂过氧化作用最小。当丙酮提取物添加量为猪油质量的0.1%时,其抗油脂过氧化活性接近添加量为0.02%茶多酚或BHT。4种提取物对DPPH自由基的清除能力大小顺序依次为:水提取物、丙酮提取物、甲醇提取物和乙酸乙酯提取物。浓度为0.0175 mg/mL的水提取物对DPPH自由基的清除率高达85.2%,高于BHT,但略低于茶多酚。石榴皮多酚是一种有效的天然抗氧化剂。     

Composition and antioxidant activity of raisin extracts obtained from various solvents

Experiments were conducted to determine if the contents of phenolics and browning reaction products and antioxidant activity of raisin extracts were closely dependent on the extraction solvent. Enhanced extraction yields were obtained from solvent containing higher water concentrations. However, total phenolic content (TPC) was highest for extracts obtained from solvent to water ratios of 60:40 (v/v), whereby the extract obtained from ethanol:water (60:40, v/v) had the highest TPC of 375 mg gallic acid equivalents (GAE)/100 g extract. HMF content was highest in extracts obtained from 60% solvent, regardless of solvent type. The extract obtained from 60% methanol had the highest HMF content at 199 μg/g extract. Although the 60% solvents provided extract with high antioxidant components, the antioxidant activity of raisin extracts obtained from 80:20 (v/v) solvent/water was significantly higher than other raisin extracts obtained from different solvent concentrations. Phenolic acids, HMF, and low-molecular-weight flavonoids were responsible for the antioxidant activity, but not the high-molecular-weight flavonoids.     

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.     

Physicochemical properties of pectins from ambarella peels (Spondias cytherea) obtained using different extraction conditions

Extraction and use of pectins from ambarella peels could add value to the waste products arising from processing of the fruit. Dried alcohol-insoluble residues (AIR) of ambarella peels were treated separately with HCl, deionised water and oxalic acid/ammonium oxalate solutions, and the resulting pectin extracts analysed for some biochemical and physicochemical parameters. The results show that pectin yield (9–30% dry AIR), uronic acid (557–727 mg/g dry weight), neutral sugars (125–158 mg/g), degree of methylation (50–58%) and acetylation (4–6%), molar mass (263,000–303,000 g/mol) and intrinsic viscosity (179–480 ml/g) varied significantly (p < 0.05) with the various extraction methods used. Extraction with oxalic acid/ammonium oxalate solution gave the highest pectin yield, with high molar mass and degree of methylation, making the extracts suitable for use as additives in the food industry. The results compared well to lime pectin extracted under the same conditions, indicating their commercial significance.     

Enhanced extraction of flavanones hesperidin and narirutin from Citrus unshiu peel using subcritical water

Flavanones including hesperidin and narirutin constitute the majority of the flavonoids that occur naturally in citrus fruits. The main purpose of this study was to extract valuable natural flavanones from agricultural by-products such as citrus peels using subcritical water extraction (SWE). Thus, the application of SWE to extraction of flavanones hesperidin and narirutin from Citrus unshiu peel was evaluated, and the effect of key operating conditions was determined by varying the extraction temperature (110–200 °C) and time (5–20 min) under high pressure (100 ± 10 atm). The maximum yields of hesperidin (72 ± 5 mg/g C. unshiu peel) and narirutin (11.7 ± 0.8 mg/g C. unshiu peel) were obtained at an extraction temperature of 160 °C for an extraction time of only 10 min. These yields accounted for approximately 99% of the total amount of these flavanones in the original material. The SWE was compared with three conventional extraction methods in terms of the extraction time and recovery yields for hesperidin and narirutin. The hesperidin yield by SWE was more than 1.9-, 3.2-, and 34.2-fold higher than those obtained by extraction methods using ethanol, methanol, or hot water, respectively, and the narirutin yield was more than 1.2-, 1.5-, and 3.7-fold higher.     

Quantification of phenolic contents and antioxidant capacity of Atlantic sea cucumber, Cucumaria frondosa

The antioxidant activity (oxygen radical absorbance capacity, ORAC) and total phenols and flavonoids were determined in extracts from digestive tract, gonads, muscles and respiratory apparatus of sea cucumber, Cucumaria frondosa. Total phenols content varied from 22.5 to 236.0 mg of gallic acid equivalents/100 g dw, and flavonoids from 2.9 to 59.8 mg of rutin equivalents/100 g. ORAC values ranged from 140 to 800 μmol of Trolox equivalents/g dw. Among all extracts, best antioxidant potencies were observed in ethyl acetate extracts from digestive tract, and in acetonitrile-rich fractions obtained from mixed extracts using acetonitrile/TFA (trifluoroacetic acid) acidified water on muscles, gonads and respiratory apparatus. The weakest potencies were observed with water extracts from digestive tract and respiratory apparatus, and with water-rich fractions obtained from mixed extraction of gonads and muscles. A significant correlation was observed between ORAC values and total phenol content in extracts and fractions of gonads and muscles, but ORAC and phenols were not correlated in digestive tract and respiratory apparatus extracts. ORAC values were significantly correlated (p < 0.05) with total flavonoids in all extracts. Successive eluates obtained from solid-phase extraction of water-rich fractions using C₁₈ cartridge showed ORAC values (105–500 μmol of TE/g) reaching up to 2.3 times the potency of their parent fractions. Flavonoids are suggested to be mainly responsible for observed activities. Our results provide a first quantitative evaluation of C. frondosa tissues as useful sources of antioxidants for human consumption.     

Raw Millefiori honey is packed full of antioxidants

Total polyphenols, flavonoids and antioxidant power of raw honey samples from two of the most common Italian varieties, i.e., Millefiori and Acacia, were evaluated. Phenolic content, expressed as caffeic acid equivalents, ranged from 12.5 to 17.5 mg/100 g and from 3 to 11 mg/100 g in Millefiori and Acacia honeys, respectively. All Millefiori samples exhibited the highest flavonoid concentration being between 1.23 and 2.93 mg catechin equivalents (CE)/100 g honey. Total flavonoids in 100 g Acacia honeys were in the range of 0.45–1.01 mg CE. Acacia honeys had lower total antioxidant power, as assessed by ferric reducing/antioxidant power assay, than Millefiori. The relationship between phenolic content and antioxidant power was discussed. Comparative experimental analysis was performed with an artificial honey and processed honeys. Raw Millefiori honey is rich in both amount and variety of antioxidant substances, and its inclusion in the diet may be recommended to complement other polyphenol sources.     

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.     

Antioxidant activity and cell toxicity of pressurised liquid extracts from 20 selected plant species in Jeju,Korea

Twenty plant species were selected from Jeju, Korea, and extracts made using a pressurised liquid extraction system. Integral antioxidative capacity (IAC), inhibitory activity of nitric oxide (NO) production, and cell toxicity were evaluated to identify new dietary antioxidants of natural origin with a low degree of cell toxicity. Geranium thunbergii and Pyrrosia lingua showed high IAC values of water-soluble substances (9.76 and 9.48 mmol ascorbic acid equivalents/g), with high IAC values of lipid-soluble substances of 5.20 and 1.97 mmol trolox equivalents/g, respectively. Prunus padus and Rhus javanica did not show any cytotoxicity on RAW 264.7 macrophages or HS-68 normal skin fibroblast cells and showed inhibitory activity of NO production (IC₅₀ = 327 and 337 μg/ml) with LPS-induced RAW 264.7 macrophages. Malus sieboldii did not exhibit cytotoxicity with either cell type and showed high inhibitory activity of NO production (IC₅₀ = 219 μg/ml) and high IAC values of water-soluble substances (4.93 mmol ascorbic acid equivalents/g).     

Comparison of aqueous extraction efficiency and biological activities of polyphenols from pomegranate peels assisted by infrared,ultrasound, pulsed electric fields and high-voltage electrical discharges

The effects of aqueous extraction of bioactive compounds from pomegranate peels using conventional extraction (CE) and extraction assisted by infrared irradiation (IR), ultrasound (US), pulsed electric fields (PEF), and high-voltage electrical discharges (HVED) have been compared. For the extractions assisted by US, PEF and HVED, the saturation in extraction was observed approximately at the same specific energy input in the order of W ≈ 90–100 kJ/kg. HVED assisted extraction enhanced the recovery of polyphenols by ≈3 and ≈1.3 times as compared to US and PEF assisted extractions, respectively. Scanning electron microscopy (SEM) data evidenced that the highest yield of total polyphenols after the HVED treatment can reflect the presence of a strong damage of the microstructure of pomegranate skins. The obtained data on inhibition of growth of A. flavus and biosynthesis of aflatoxin B1 were explained accounting for the presence of different synergetic effects of phenolic compounds on inhibition of different bioactivities. All the studied extracts (0.2 mg/mL) demonstrated the higher inhibition efficiency for S. aureus (up to ≈80%) as compared to E. coli (up to ≈33%). PEF selectively extracted and enhanced the recovery of ellagic acid (≈740 μg/g DM), whereas HVED (≈345 μg/g DM) intensified gallic acid extraction compared to US, IR, HVED and WB.     

Antioxidant capacity and mineral content of pulp and peel from commercial cultivars of citrus from Brazil

Four Citrus species (C. sinensis, cvs. Pera and Lima; C. latifolia Tanaka cv. Tahiti; C. limettioides Tanaka cv. Sweet lime and C. reticulate, cv. Ponkan) grown in Brazil were characterised in relation to contents of minerals, ascorbic acid, total polyphenols and antioxidant capacity of pulps and peels. In general, the peels demonstrated significantly higher contents of all compounds than the pulps (p < 0.05), with the exception of the Pera orange pulp that presented the highest acid ascorbic content (68 mg/100 ml), while the Tahiti lime peel presented the lowest (8 mg/100 g). Citrus showed high levels of potassium, calcium and magnesium, and the peels were considered sources of these minerals. The Ponkan mandarin peel presented the highest antioxidant capacity. The antioxidant capacity of citrus was correlated both to vitamin C and phenolics. Aside from citrus pulps, the peels are also good sources of bioactive compounds and minerals, and can be explored for their health promoting values in food products.     

Effects of various solvents on the extraction of antioxidant phenolics from the leaves, seeds, veins and skins of Tamarindus indica L.

The effects of solvents, of varying polarities, on the extraction of antioxidant phenolics from the leaves, seeds, veins and skins of Tamarindus indica (T. indica) were studied. The efficiencies of the solvents for extraction of the antioxidant phenolics were in the order: methanol > ethyl acetate > hexane. Phenolic content ranged from 3.17 to 309 mg of gallic acid equivalents/g. Methanol leaf extract (MEL) had the highest phenolic content and was the most potent scavenger of DPPH and superoxide radicals. Methanol vein extract had the highest ferric reducing activity whereas methanol seed extract was the most potent ABTS radical-scavenger. A positive correlation existed between phenolic content and antioxidant activities of the plant parts. HPLC analyses of MEL revealed the presence of catechin, epicatechin, quercetin and isorhamnetin. Overall, methanol was the most effective solvent for extraction of antioxidant phenolics from T. indica. T. indica, particularly the leaf, can be a useful source of natural antioxidants.     

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.