A NEW METHOD FOR ELLAGIC ACID PRODUCTION FROM POMEGRANATE HUSK

Pomegranate ( Punica granatum L.) husk, a by-product of the pomegranate juice industry, is an inexpensive and abundant source of ellagic acid. Ellagic acid is widely used as functional food for its physiological functions. It is the breakdown product of ellagitannins. To date, the preparation of ellagic acid from pomegranate husk has not been reported. This article reports a new process for ellagic acid production from pomegranate husk by extraction of tannins followed by acid hydrolysis and purification by extraction and recrystallization. Several tests were conducted to obtain optimum conditions including extraction of tannins by varying solvents, acid concentration and reaction time for acid hydrolysis and the volume of methanol used for purification. Ellagic acid (3.5 g) with 90% purity from 100 g pomegranate husk was obtained. This new method is easy to scale up. All equipment used in this production process is widely used in food processing industry. The cost of production is low. It is suitable for industrial applications.

PRACTICAL APPLICATIONS

The production of ellagic acid is easier and the yield and purity of ellagic acid produced this way are higher than before. This method can be used not only for experiment in laboratory but also for industrial applications. The material – pomegranate husk – is a by-product of the pomegranate juice industry, so it is very cheap and easy to get. High-purity ellagic acid produced this way is sold to many companies back home and abroad. It is used as food additive and cosmetic material because of its antioxidant activity and whiteningfunction. The toxicity of pomegranate husk is lower than that of gallnut, which has been the main material of ellagic acid production in the past. Reagents are common and inexpensive; some of them are reusable.     

A survey of ellagitannin content in raspberry and blackberry cultivars grown in Trentino (Italy)

This study describes the first application of a new procedure for acid hydrolysis of Rubus ellagitannins in methanol, which allows quantification of all the major reaction products (ellagic acid, methyl-sanguisorboate, methyl gallate and an unknown ellagic acid derivative) and estimation of the mean degree of polymerization (mDP) of Rubus ellagitannins. The ellagitannins of 21 raspberry and 15 blackberry cultivars produced in Trentino, Italy during 2005 were extracted and characterised. The total content of ellagitannins was shown to vary by a factor of 2 among raspberry cultivars and by a factor of 4 among blackberry cultivars.     

响应面法优化树莓鞣花酸提取工艺及其体外抗氧化活性

本研究旨在通过微波辅助提取法确定树莓鞣花酸的最佳提取工艺,并对其抗氧化活性进行研究。以红树莓为实验原料,使用乙醇提取溶剂,采用微波辅助溶剂法提取红树莓中鞣花酸,通过单因素试验与响应面试验优化红树莓中鞣花酸提取工艺,利用高效液相色谱(HPLC)法对鞣花酸进行分析测定,并对其抗氧化性进行研究。结果表明:响应面试验优化出最佳提取条件为:提取时间1.8 min、提取温度60 ℃、乙醇浓度50%、液料比19:1、微波功率411 W,此条件下测得鞣花酸提取量为(82.75±1.05) mg/100 g,高于传统盐酸水解法在最佳提取条件下所测树莓鞣花酸提取量。实际结果与理论预测高度吻合,因此本实验得到的最优条件准确可靠有参考应用价值。微波辅助溶剂法所得鞣花酸对羟自由基清除能力、清除DPPH自由基能力都显著高于传统盐酸水解法所得鞣花酸对羟自由基清除能力,清除DPPH自由基能力,说明微波辅助溶剂提取法,不但提取时间短暂,提取溶剂用量少,而且更好的保护了树莓鞣花酸的抗氧化活性,可为树莓鞣花酸工业生产提供一定参考。     

刺梨中鞣花酸的分离鉴定及提取工艺的优化

目的：本研究旨在建立刺梨中鞣花酸的分离纯化方法,并对提取鞣花酸的酸水解法进行工艺优化。方法：以刺梨干果为原料,通过超声辅助提取法提取单宁酸,加酸进行水解制得粗鞣花酸,并通过甲醇重结晶进行纯化,采用高效液相色谱测定其纯度,及核磁共振波谱法(NMR)、红外光谱法(IR)、质谱法(MS)、高效液相色谱法(HPLC)、紫外光谱法(UV)对提取得到的鞣花酸进行结构确证,另外,以得率为指标,以酸浓度、提取温度、提取时间、液料比为自变量,在单因素实验的基础上,通过Box-Benhnken中心组合设计和响应面分析法确定刺梨鞣花酸的酸水解工艺条件。结果：从刺梨中提取得到了鞣花酸提取物,通过HPLC分析测定其纯度达到92%,鞣花酸提取物的最大吸收波长254 nm,分子离子峰m/z 301.00,红外吸收各波数与标准品一致,并利用¹H-NMR,¹³C-NMR确定了其结构。通过响应面法得到了酸水解法制取鞣花酸的最优工艺：酸浓度5%,提取温度127℃,液料比为8:1 mL/g,提取时间9.8 h,在此条件下刺梨鞣花酸的得率为30.2%±2.3%(与预测值相比P>...     

Potential dietary sources of ellagic acid and other antioxidants among fruits consumed in Brazil: jabuticaba (Myrciaria jaboticaba (Vell.) Berg)

BACKGROUND: The objectives of this study were (1) to evaluate the content of ellagic acid in fruits consumed by the Brazilian population, including native ones; (2) to further characterize rich sources in relation to ascorbic acid, phenolics contents and in vitro antioxidant capacity; and (3) to study the distribution and effect of ripening stage on ellagitannins content of jabuticaba (Myrciaria jaboticaba). The content of free ellagic acid and ellagic acid derivatives such as ellagitannins was analyzed using high‐performance liquid chromatography (HPLC). RESULTS: Ellagic acid was detected in 10 out of a total of 35 fruits analyzed. The content of free ellagic acid in fruits varied from 0.0028 to 0.085 g kg^?1 (FW) and total ellagic acid varied from 0.215 to 3.11 g kg^?1 (FW). All the seven fruits belonging to the Myrtaceae family evaluated in this study presented high contents of ellagitannins in their composition, with jabuticaba, grumixama and cambuci (all native from Brazil) showing the highest total ellagic acid contents. Jabuticaba, the most consumed in Brazil among those and already adapted to commercial plantations, contained concentrated phenolics compounds, including ellagitannins, in the peel. Anthocyanins (cyanidin derivatives) increased significantly through ripening of jabuticaba and were not present in the pulp or seeds. Samples collected from three different locations during summer, winter and spring had total ellagic contents varying from 1.88 to 3.31 g kg^?1 (FW). The decrease in ellagic acid content with ripening was more accentuated for pulp (eight times) compared to seeds (2.3 times) and peel (2.0 times). CONCLUSION: These results showed the potential of jabuticaba as dietary source of ellagic acid and reinforced consumption of the whole fruit by the population. Copyright © 2011 Society of Chemical Industry     

Antioxidant compounds and antioxidant capacity of Peruvian camu camu (Myrciaria dubia (H.B.K.) McVaugh) fruit at different maturity stages

The antioxidant capacities of ascorbic acid and phenolic compounds present in camu camu fruit were screened during ripening. Ascorbic acid decreased, and anthocyanin, flavonol and flavanol contents, and DPPH antioxidant capacity increased during ripening. Antioxidant compounds from camu camu were fractionated in two fractions: an ascorbic acid-rich fraction (F-I) and a phenolics-rich fraction (F-II). F-I was the major contributor to the DPPH antioxidant capacity (67.5–79.3%) and F-II played a minor role (20.7–32.5%). A total of 30 different phenolic compounds were detected by HPLC-PAD. The presence of catechin, delphinidin 3-glucoside, cyanidin 3-glucoside, ellagic acid and rutin was elucidated. Other phenolic compounds, such as flavan-3-ol, flavonol, flavanone and ellagic acid derivatives, were also present. For the three ripening stages the flavan-3-ols and ellagic acid group were the most representative phenolic compounds in this fruit. Acid hydrolysis of F-II revealed the presence mainly of gallic and ellagic acids, suggesting that camu camu fruit possesses important quantities of hydrolysed tannins (gallo- and/or ellagitannins). These results confirm that camu camu fruit is a promising source of antioxidant phenolics.     

Antioxidant capacity and phenolic composition of different woods used in cooperage

This study has investigated the antioxidant capacity of different woods used in cooperage and the effect of the botanical species of wood on this capacity. Quercus robur and Castanea sativa were the species with the highest antioxidant capacity, due to their relatively high content of phenolic compounds. However, the phenolic content of Prunus avium samples was very low, also reflected in its antioxidant capacity. All measurements of antioxidant capacity were consistent with the content and composition of the phenolic compounds detected in the wood samples. The major contributors to the antioxidant capacity were identified as phenolic acids, including gallic, protocatechuic, p-coumaric and ellagic acid and all the ellagitannins, due to their characteristic structure.     

Antioxidant capacity, scavenger activity, and ellagitannins content from commercial oak pieces used in winemaking

In 2005, the European Union authorized the use of oak wood pieces in winemaking, and consequently, the addition of oak chips has been generalized. Thus, the antioxidant capacity, scavenger activity, and ellagitannins content from fourteen different commercial oak wood pieces (Quercus spp.) that can be found on the Portuguese market as alternatives to barrels for aging wines were studied. Commercial oak wood pieces samples from Quercus petraea had significantly higher antioxidant activity values than the pieces from Quercus alba species. In general, antioxidant capacity and scavenging activity decrease with the toasting level of the oak wood pieces samples analyzed. A similar effect was detected in the individual ellagitannin content studied. On the other hand, with the toasting process, the total phenolic content and the ellagic acid concentration increased. Finally, a correlation analysis revealed that ellagitannins contribute to the antioxidant capacity of oak wood pieces studied. From the data presented in this paper, antioxidant capacity and ellagitannin content of the oak wood pieces studied have been clearly related either to the oak wood species used, particle size, or to toasting levels.     

Identification of Ellagic Acid in Blackberry Juice Sediment

ABSTRACT: The sediment in a commercial reconstituted 'Evergreen' blackberry ( Rubus laciniatus L.) juice concentrate was found to be composed of ellagic acid, protein, and other unidentified compounds. The qualitative tannin and protein-tannin haze test indicated that the sediment was predominantly tannin or protein-tannin complexes. Nitrogen determination showed the sediment to be 6.69%± 2.21% protein on a dry-weight basis. Almost all of the extractable material was identified as ellagic acid by high-performance liquid chromatography and liquid chromatography-mass spectrometry. The ellagic acid content of the wet sediment was 0.05 g/100 g, whereas it was 7.41 g/100 g in freeze-dried sediment. Tannase enzyme did not significantly decrease the concentration of ellagitannins in Marion blackberry ( Rubus spp. Hyb.) juice in this study.     

Extraction and determination of ellagic acid contentin chestnut bark and fruit

Chestnuts are an important economic resource in the chestnut growing regions, not only for the fruit, but also for the wood. The content of ellagic acid (EA), a naturally occurring inhibitor of carcinogenesis, was determined in chestnut fruits and bark. EA was extracted with methanol and free ellagic acid was determined by HPLC with UV detection, both in the crude extract and after hydrolysis. The concentration of EA was generally increased after hydrolysis due to the presence of ellagitannins in the crude extract. The concentration varied between 0.71 and 21.6 mg g⁻¹ (d.w.) in un-hydrolyzed samples, and between 2.83 and 18.4 mg g⁻¹ (d.w.) in hydrolyzed samples. In chestnut fruits, traces of EA were present in the seed, with higher concentrations in the pellicle and pericarp. However, all fruit tissues had lower concentrations of EA than had the bark. The concentration of EA in the hydrolyzed samples showed a non-linear correlation with the concentration in the unhydrolyzed extracts.     

鞣花酸代谢产物——尿石素的研究进展

鞣花单宁和鞣花酸是广泛分布于多种蔬菜、水果和坚果中的多酚类抗氧化剂,其在胃肠道的吸收较差,生物利用率较低,但部分会被哺乳动物肠道微生物转化为更易吸收的尿石素类代谢产物。因此,人们推测这些代谢产物可能为鞣花单宁及鞣花酸发挥生物学活性的最终物质基础。本文对近年来国内外有关尿石素化学性质和抗氧化、抗炎、抗癌以及调节肠道菌群等生物活性的研究进行综述,以期为富含鞣花单宁及鞣花酸类食品的科学利用提供借鉴。     

Microwave-assisted extraction of phenolic compounds from wine lees and spray-drying of the extract

Most research on extraction of phenol compounds from wine by-products and commercial exploitation of extracts use grape seeds and/or skins as raw materials. Looking for alternative antioxidants sources, obtaining antioxidant extracts from wine lees (also known as dregs), a sub-exploited by-product of winemaking process, is here presented. Microwave-assisted extraction (MAE) of phenolic compounds from wine lees has been optimized using the total phenols index, the ORAC values and yield of the extraction as response variables. Under the optimal working conditions, the proposed MAE method provides better extraction efficiency in a much shorter time (17 min) than the conventional extraction method for phenolic compounds (24 h). The liquid extract obtained by MAE was spray-dried. The type and amount of excipients used, as well as the spray-drying temperature, were optimized in order to minimize the oxidation of phenolic compounds and maximize the yield of the spray-drying process. The total phenols index in the dried extract thus obtained was 36.8% (expressed as gallic acid), showing an ORAC value of 3930 μmol TE/g. Additionally, Mv3G, Cm-Mv3G, myricetin, quercetin, quercetin-3-β-glucoside, caffeic acid and p-coumaric acid were quantified in the dry extract by HPLC–DAD. The results indicate that wine lees antioxidant extracts can be a suitable and cheap alternative to those obtained from grape seeds or skins.     

Pomegranate (Punica granatum) supplements: Authenticity,antioxidant and polyphenol composition

Pomegranates contain a complex mixture of gallotannins, ellagitannins, ellagic acid and anthocyanins. However, label claims on pomegranate supplements (PS) may not correlate with actual content of antioxidants, polyphenols or tannins. Nineteen PS were evaluated for their authenticity by determining ellagitannin composition by RP-HPLC and studying the relationship between total polyphenols as measured by the Folin–Ciocalteau assay and antioxidant capacity by oxygen radical absorbing capacity (ORAC), free radical scavenging properties by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and ferric reducing antioxidant power (FRAP). Only a limited number of pomegranate supplements were authentic. Product labels were inconsistent with polyphenol composition and antioxidant content. A majority of the samples (n = 13) contained disproportionately high amounts of ellagic acid and low or no detectable pomegranate tannins. Only six products had tannin composition that resembled pomegranates (punicalagin, punicalin, ellagitannins and gallotannins). PS-01 (natural pomegranate extract) was the most representative of pomegranate fruit polyphenols with 99% total pomegranate polyphenol and the highest antioxidant capacity across all measures. Correlations between total polyphenols and antioxidant content were high (R² > 0.87) in products that had polyphenol composition resembling pomegranates. Products that contained high amounts of ellagic acid and low or no detectable pomegranate tannins had poor correlations between total polyphenols and antioxidant content. The results indicate that reliable labeling information, better standardization, improved manufacturing practices and regulation of the market is required to assure consumers of the quality of pomegranate supplements.     

Fabrication of nanoparticles using partially purified pomegranate ellagitannins and gelatin and their apoptotic effects

Scope: Nanoparticles possess unique chemical and biological properties compared to bulk materials. Bioactive food components encapsulated in nanoparticles may have increased bioavailability and bioactivities. Methods and results: Self‐assembled nanoparticles made of partially purified pomegranate ellagitannins (PPE) and gelatin were fabricated using three PPE‐to‐gelatin mass ratios (1:5, 5:5, and 7:5). The PPE contained 16.6% (w/w) of punicalagin A, 32.5% (w/w) of punicalagin B, and a small amount of ellagic acid‐hexoside and ellagic acid (1%, w/w). Nanoparticles fabricated using the ratio 5:5 had a particle size of 149.3±1.8 nm, positive zeta‐potential of 17.8±0.9 mV, production efficiency 53.0±4.2%, and spherical morphology under scanning electron microscopy. Loading efficiency of punicalagin A and punicalagin B in these particles were 94.2±0.4% and 83.8±0.5 %, respectively. Loading capacity was 14.8±1.5% and 25.7±2.2%, respectively. Only punicalagin anomers were able to bind with gelatin to form nanoparticles, whereas ellagic acid‐hexoside or ellagic acid could not. Fourier transform infrared spectroscopy suggested that the interactions between ellagitannins and gelatin were hydrogen bonding and hydrophobic interactions. PPE‐gelatin nanoparticle suspension was less effective than PPE in inducing the early stage of apoptosis on human promyelocytic leukemia cells HL‐60. But they had similar effects in inducing late stage of apoptosis and necrosis. Conclusion: Pomegranate ellagitannins bind with gelatin to form self‐assembled nanoparticles. Ellagitannins encapsulated in nanoparticles had decreased apoptotic effects on leukemia cells HL‐60.     

An in vitro method to simulate phenolic compound release from the food matrix in the gastrointestinal tract

A method to simulate in vitro the physiological conditions in the stomach and small intestine has been developed as a modification of a method already described by Miller et al. [11]. This method enables the study of phenolic compound release from the food matrix and their transformation during digestion, and has been applied to orange juice, strawberries, and strawberry jam. The dialysis rates obtained for the different phenolics in the new method are much better than those found in the original one. Some food constituents, as is the case of sugars in jams, can affect the dialysis rates in vitro, showing that the results obtained for dialysis should be taken with caution in these products. These assays have shown that strawberry anthocyanins are largely degraded (transformed) during the digestion and that free ellagic acid is released from ellagitannins leading to a ten-fold increase of this compound during digestion.     

Development of Advance Extraction Methods for the Extraction of Myristicin from <Emphasis Type="Italic">Myristica fragrans</Emphasis>

Myristicin in a psychoactive, hallucinogenic substance and as a constituent of nutmeg and mace (Myristica fragrans nut and its aril, respectively) is widely available for people seeking so-called legal highs of natural origin. In this work, we present three methods for determination of myristicin from plant material. The methods consist of optimized extraction procedures: microwave-assisted extraction, ultrasound-assisted extraction utilizing ultrasound bath or sonotrode and gas chromatography-mass spectrometry (GC-MS) analysis method. The methods were validated: repeatability and intermediate were below 10 % and recovery ranged from 97 to 109 %. The developed methods were then used for determination of myristicin in commercially available nutmeg and mace samples. The concentration ranged from 0.58 to 12.94 μg mg^?1, and the highest concentration of myristicin was observed in samples of those producers, which are often recommended by individuals experimenting with nutmeg. The microwave-assisted extraction (MAE) method has been proven to be the most effective one for the extraction of myristicin from plant material, as it also generates deeper changes in sample structure than UAE techniques. The MAE/GC-MS method was successfully used for nutmeg residue analysis, allowing identification of five characteristic violate constituents of nutmeg.     

微波辅助萃取技术研究进展

微波萃取技术是近年来发展较快的一种新型提取技术，与传统的天然药物活性成分提取方式相比，具有方便、省时、能耗少、有效成分得率高、选择性强和适于工业化生产等优点，因而越来越得到相关领域研究人员的关注。作者阐述了微波萃取技术在天然产物活性成分提取中的研究现状以及发展前景。     

Extraction of phenolics from new oak casks during spirit maturation: impact on spirit colour

The increase in colour and solubilisation of extract, polyphenols and ellagitannins were studied in wine spirit in new oak casks over a period of 23 years. Ellagitannins and oak colour were solubilised during the early years of maturation. After five years, the concentration of ellagitannins decreased markedly, while colour increased, although at a lower rate. Ellagitannins were oxidised into coloured polymers and hydrolysed into ellagic acid. The dry extract, containing polysaccharides, polyphenols and lignin, increased throughout maturation, growing quickly during the early years and slowly thereafter. In a model solution, ethanol exhibits a strong solvent effect. For higher concentrations (50% v/v and more), the extraction of colour was faster, and was typically dark yellow/brown with a red tone, attributable in part to the solubilisation of lignin and lignin by-products. © 2020 The Institute of Brewing & Distilling     

Structural modification by different pretreatment methods to enhance microwave-assisted extraction of β-carotene from carrots

Since the structure of a material is a key factor influencing the extraction efficiency, any means to modify the structure to enhance extraction is attractive. As limited information is available on the effect of sample pretreatment prior to extraction on the extractability of a bioactive compound, the effects of selected pretreatment methods, i.e., soaking in citric acid, blanching in water as well as in citric acid, on the sample structure and subsequent microwave-assisted extraction (MAE) were investigated; carrots and β-carotene (as well as carotenoids) were selected as the test material and bioactive compound, respectively. A suitable condition to extract β-carotene in terms of the microwave power and extraction time was determined. Comparison between MAE and Soxhlet extraction was also made. At the optimized condition the contents of β-carotene and total carotenoids extractable from carrots blanched in water (29.74 and 58.04 mg/100 g dry basis) and in citric acid (32.08 and 61.62 mg/100 g dry basis) were significantly higher than those from the untreated carrots (23.26 and 51.79 mg/100 g dry basis). The antioxidant activities of the extracts obtained from carrots blanched in water and in citric acid were also higher than that from carrots with no pretreatment. Comparison between MAE and Soxhlet extraction revealed that the extract from MAE contained lower amounts of β-carotene and total carotenoids and exhibited lower antioxidant activity; the required MAE time was significantly shorter, however.     

Microwave-assisted extraction of sulforaphane from white cabbages: Effects of extraction condition,solvent and sample pretreatment

Sulforaphane is one of the most potent food-derived anticarcinogens and is abundantly found in Brassica vegetables, including cabbages. Due to its importance an ability to effectively extract sulforaphane from its source is desirable. This work investigated the feasibility of using microwave-assisted extraction (MAE) to extract sulforaphane from cabbages. The effects of various parameters, which are the type of solvent (dichloromethane and water), set microwave power (130, 260 and 390 W) and extraction time, on the extraction yield of sulforaphane were investigated. In addition, since it was previously shown that partially dried cabbages contain higher sulforaphane content than fresh cabbages, the effect of partial drying prior to MAE on the extractable amount of sulforaphane was investigated. A comparison between MAE and conventional extraction was also made. The results showed that MAE was more effective than the conventional extraction as the former led to higher yield of sulforaphane in a much shorter extraction time. There were no significant differences in the highest extractable sulforaphane contents from the fresh and semi-dried samples. Use of different solvents also did not lead to any significant effect on the MAE of sulforaphane, either from the fresh or semi-dried sample.