Correlation between antioxidant concentrations and activities of Yuja (Citrus junos Sieb ex Tanaka) and other citrus fruit

The antioxidant concentrations and activities of tangerines, yuja, lemons, and oranges in the Korean marketplace have been investigated. Flesh and peel tissues of the fruit were analyzed. The total vitamin C, total phenolics, flavonoid, hesperidin, and total antioxidant activity of all citrus types were higher in peel than flesh tissues. Total phenolic concentrations of flesh tissues were in the following order: lemon>yuja>tangerine>orange, while those concentrations in the peel were tangerine >yuja>orange>lemon. Lemon and yuja had higher hesperidin concentrations in flesh and peel tissues, respectively. Among the flesh samples, the total flavonoid and total phenolic concentrations were highly related with total antioxidant activity. The relationship between flavonoid and hesperidin was also strong among flesh and peel tissues. Our results indicate that citrus fruit includes high antioxidant compounds and activity and that citrus peel could be a good source of food ingredients.     

索氏法与超声波辅助提取甜橙橙皮苷比较研究

为了确定从甜橙果皮中提取橙皮苷的高效方法,比较了索氏法和超声波辅助提取的效果。考虑到不同提取条件,即溶剂浓度、液固比、提取时间、提取次数和超声波功率的影响,采用单因素及正交设计比较这两种方法提取橙皮苷的效果,并用RP-HPLC法测定提取液中橙皮苷的含量。结果表明,超声波法辅助提取3个甜橙品种果皮中橙皮苷的平均含量高于索氏提取法的26.5%～34.6%,用这两种方法提取橙皮苷的数量达到统计学上的显著差异。超声波辅助提取法能真实地反映甜橙果皮中橙皮苷的含量,具有操作简便、省时的优点。建议柑橘果实橙皮苷的测定和提取可采用超声波辅助法。     

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.     

Important flavonoids and limonin in selected Thai citrus residues

The distributions of important flavonoids and limonin in selected Thai citrus residues were investigated in this study. The residues of interest were peels of pomelo (Citrus grandis (L.) Osbeck cv. Kao Yai and cv. Kao Nampheung), residues after juice extraction of tangerine (Citrus reticulata Blanco cv. Sainamphueng and cv. Bangmod) and peels and residues after juice extraction of kaffir lime (Citrus hystrix DC). Flavonoids were characterized and quantified in terms of flavanones, i.e., naringin and hesperidin, and polymethoxyflavones (PMFs), i.e., nobiletin, sinensetin and tangeretin. Naringin was a dominant flavanone in polmelo peels. Three PMFs were found in all samples with varying contents. Comparing the samples, residues from tangerine cv. Bangmod exhibited the highest value of each PMFs. Limonin was present in small amounts in pomelo peels and residues after juice extraction of tangerine and kaffir lime. All samples were good sources of dietary fibre, with a total dietary fibre content of more than 60 g/100 g on a dry weight basis with high proportion of soluble dietary fibre.     

超声技术在橘皮中提取橙皮苷的应用

研究了橘皮中类黄酮化合物——橙皮苷的超声提取方法。在超声波作用下，以甲醇为溶剂，从橘皮中提取橙皮苷，通过对甲醇的浓度、提取时间、料液比和橘皮粉碎的粒度等因素进行的研究，确定了提取溶剂为甲醇，最佳的提取条件为液料比=50mL：1g，提取时间30min，橘皮的粒度为0.45～0.08mm。实验的重复性和回收率也较好(HPLC检测)。     

Thermomechanical process intensification for oil extraction from orange peels

The study investigated the intensification and improvement of oil extraction from orange peel through a thermomechanical process: the instantaneous controlled pressure drop (briefly D.I.C. process). This process involves subjecting orange peel for a short time to steam pressure, followed by an instantaneous decompression to a vacuum at 50 mbar. Central composite design was used to study the combined effects of processing steam pressure (1–7 bar; which corresponds to a temperature ranging between 100 °C and 162 °C respectively), processing time (0.3–3.7 min) and initial moisture content of orange peel before thermomechanical oil extraction (9.8–60.2% on dry material basis). The quantitative analysis, have been undertaken on oil present in orange peels, after processing. Correlation analysis of the mathematical regression model indicated that quadratic polynomial model could be employed to optimize the extraction of oil from orange peel. From response surface plots, the three variables exhibited a linear effect with the strongest effect for the processing pressure. The optimum reaction conditions selected with response surface analysis were as follows: steam processing pressure: 6.6 bar, processing time: 3.68 min, initial moisture content: 53.6% d.m. basis. Under these conditions, experimental yield of orange peel oil was close to predicted value (99%) calculated from the polynomial response surface model equation. A kinetic study indicated that extraction performed by D.I.C. process is clearly quicker than conventional steam distillation method.

Industrial relevance

World production of citrus fruit has experienced continuous growth in the last decades of the XX century. Total annual citrus production was estimated at over 105 million tons in the period 2000–2004 (http://www.unctad.org/infocomm/anglais/orange/market.htm). Oranges constitute the bulk of citrus fruit production, accounting for more than half of global citrus production in 2004. Thus, the quantities of generated by-products are very important. In this study, extraction of oil from orange peel was performed through a thermomechanical process developed in our laboratory. We report on some results obtained using this process as well as others comparing extraction kinetics when using a conventional steam distillation process and the proposed extraction process. One of the advantages of the described method is the short contact of extracted oil with the heated zones in the apparatus resulting in a lower heating period of the product compared to steam distillation. At industrial scale, this is interesting regarding oil quality and energy saving.     

Improving the pressing extraction of polyphenols of orange peel by pulsed electric fields

The influence of pulsed electric field (PEF) treatment on the extraction by pressing of total polyphenols and flavonoids (naringin and hesperin) from orange peel was investigated. A treatment time of 60 μs (20 pulses of 3 μs) achieved the highest cell disintegration index (Zp) at the different electric field strengths tested. After 30 min of pressurization at 5 bars, the total polyphenol extraction yield (TPEY) increased 20%, 129%, 153% and 159% for orange peel PEF treated at 1, 3, 5 and 7 kV/cm, respectively. A PEF treatment of 5 kV/cm to the orange peels increased the quantity of naringin and hesperidin in the extract of 100 g of orange peels from 1 to 3.1 mg/100 g of fresh weigh (fw) orange peel and from 1.3 to 4.6 mg100 g fw orange peel respectively. Compared to the untreated sample, PEF treatments of 1, 3, 5 and 7 kV/cm increased the antioxidant activity of the extract 51%, 94%, 148% and 192%, respectively.The results of this investigation demonstrate the potential of PEF as a gentle technology to improve the extraction by pressing of polyphenols from fresh orange peel. This procedure enhances the antioxidant capacity of the extracts, reduces extraction times and does not require using organic solvents.Industrial relevanceProcessing of orange fruits to obtain fresh juice or citrus-based drinks generates very large amounts of byproduct wastes, such as peels that are a rich source of polyphenols mainly flavonoids. Extraction of these compounds from orange peels is a crucial step for use of these compounds in the food and pharmaceutical industries as antioxidants. PEF-assisted extraction by pressing of polyphenols from fresh orange peels stands as an economical and environmentally friendly alternative to conventional extraction methods which require the product to be dried, use large amounts of organic solvents and need long extraction times.     

Functional orange juice enriched with encapsulated polyphenolic extract of lime waste and hesperidin

Polyphenolic extract from lime waste and hesperidin were encapsulated to reduce the bitter taste of flavonoid glycosides, mitigate destruction of nutrients during heat process and increase the nutrition value of orange juice. The properties of polyphenolic extract of lime waste, encapsules, the physicochemical and the sensory factors of enriched orange juice during storage were measured. The value of yield of the process (Y) and encapsulation efficiency (EE) was obtained within the range of 85–90% and 59–71%, respectively. The particle size of samples was reported within the range of 27–41 μm. The physicochemical properties in treatments follow the same pattern when compared with the control sample in 60‐day storage. In the sensory assessment, encapsulated formulas were considered to have the best treatment. If the hesperidin was not encapsulated, 80% of it would have been destroyed during pasteurisation, while when encapsulated, only 18.8–33.18% of hesperidin was destroyed.     

Bioactive compounds extracted by liquid and supercritical carbon dioxide from citrus peels

This work investigated the extraction of bioactive compounds from citrus peels, an agri-food waste. Carbon dioxide (CO₂), an eco-friendly solvent, was used under liquid and supercritical conditions to perform the extractions from orange, tangerine and lemon peels. The possibility of using ethanol as a cosolvent at small percentages up to 20% was also studied. The extraction yield, total polyphenolic content, individual polyphenolic profile, antiradical activity and volatile organic compounds of the extracts were evaluated. The highest yields were obtained when 20% ethanol was used as a cosolvent in both liquid (at 20 MPa and 20 °C) and supercritical (at 30 MPa and 60 °C) CO₂ extraction. In addition, the extracts obtained with liquid CO₂ + 20% ethanol showed the highest content of naringin (35.26, 44.05 and 19.86 mg g^-1 in orange, tangerine and lemon peel extracts, respectively) and terpenes, in particular limonene. This type of extract also showed the highest antiradical activity (31.78–59.51 µmolTE g^-1) as measured by both ABTS·+ and DPPH·. These findings show that the extraction with a liquid CO₂ and ethanol mixture could be a valid alternative to traditional solvent extraction using 80% less organic solvent and producing extracts with high antiradical capacity and rich in volatile organic compounds.     

14种柑橘皮中多甲氧基黄酮成分分析

建立柑橘皮中9 种多甲氧基黄酮（polymethoxyflavonoids,PMFs）快速定量检测方法,比较不同品种来源柑橘皮中PMFs的含量差异,为寻找高含量PMFs高生物活性柑橘皮的药食两用素材提供参考。采用超声乙醇提取、高效液相色谱方法快速准确检测PMFs,并对14 种柑橘皮、陈皮中9 种特征物质PMFs含量进行比较。结果显示9种PMFs在对照品溶液质量浓度范围内具有良好的线性关系,且准确度和精确度良好。分析发现,不同品种来源柑橘皮中PMFs种类和含量差异很大。江西陈皮、重庆柑橘、广西皇帝柑、广西砂糖橘果皮中PMFs总含量很高,均在7495.19 μg/g以上,其中广西沙糖橘果皮PMFs总含量高达10 412.18 μg/g。该方法简便,稳定性好,可为柑橘皮中PMFs生物活性研究的深入、柑橘皮产品开发和中药原料选材及其质量控制提供科学依据。     

CITRUS PRODUCT COLOR ENHANCEMENT USING EXTRACTS OF PEEL OF DIFFERENT VARIETIES

SUMMARY— Colored fractions ware extracted and concentrated from peel of most commercial orange varieties and used to enhance the color of orange juice products. These included Pineapple, Hamlin, Parson Brown, Valencia, blood orange and Dancy tangerine. The relative yields of crude pigment per kg of peel were compared between he different varieties. The color extracts were analyzed for tristimulus yellowness (Y) and redness (RI factors. Crude pigment yields ranged from .34 g (Hamlin) to 1.29 g (Dancy tangerine) per kg of peel. R factors were highest for Dancy tangerine, followed by Pineapple orange and Valencia, and lowest for Hamlin. Y factors were highest for Dancy tangerine, followed by Valencia and Pineapple and lowest for Hamlin. Most color extracts were effective at a dilution of 113500 parts in improving the color of orange juice products.     

Flavanone glycosides in Brazilian orange juice

Authentic samples of oranges, frozen concentrated orange juice and pulp-wash, and retail samples of freshly squeezed orange juice and frozen concentrated orange juice have been collected in Brazil and analysed for the flavanone glycosides (FG) narirutin and hesperidin by reversed phase HPLC with UV detection at 280 nm. The juice from hand-squeezed fruit gave narirutin and hesperidin concentrations of 16–142 mg l⁻¹ and 104–537 mg l⁻¹, respectively. The ratio of hesperidin to narirutin showed varietal difference with Pera having the highest ratio (mean 8.4) and Baı́a the lowest (3.6). Frozen concentrated orange juice contained higher quantities of FG with narirutin ranging from 62 to 84 mg l⁻¹ and hesperidin from 531 to 690 mg l⁻¹ (after dilution to 12 °Brix). In frozen concentrated orange juice pulp-wash, the narirutin level ranged from 155 to 239 mg l⁻¹ and hesperidin from 1089 to 1200 mg l⁻¹. The analysis of 23 samples of freshly squeezed juice from the Brazilian market place showed that the FG content of most samples (9.1 to 94.8 and 105.8 to 586.6 mg l⁻¹, respectively, for narirutin and hesperidin) was similar to those found for authentic ones, indicating that these orange juices were not adulterated.     

Chemical composition and biological activity of Citrus jambhiri Lush

The fresh peel of Citrus jambhiri was extracted with aqueous methanol and the residue was fractionated using light petroleum, chloroform and ethyl acetate. The constituents of the extracts were separated by column chromatography employing solvents of different polarity. The chemical structure of the isolated compounds was then identified by MS and NMR. Column chromatography of the petroleum fraction resulted in the isolation of nobiletin, 5-O-demethylnobiletin, tangeretin, 5-hydroxy-3,6,7,8,3′,4′-hexamethoxyflavone, 3,5,6,7,8,3′,4′-heptamethoxyflavone, and a mixture of β-sitosterol and stigmasterol. The chloroform fraction afforded 6-demethoxynobiletin, 5,4′-dihydroxy-6,7,8,3′-tetramethoxyflavone, limonin and nomilin. The flavonoid glycosides naringin, hesperidin and neohesperidin were isolated from the ethyl acetate fraction. The chemical structure of the isolated compounds was established by MS and NMR (APT, COSY, HSQC, HMBC, and NOESY). LC–ESI-MS analysis of the ethyl acetate fraction afforded eight flavonoid glycosides, while the dichloromethane fraction of the defatted seeds contained seven limonoid aglycones. The chloroform fraction exerted the strongest DPPH^∗ free radical scavenging activity in comparison to other fractions. The petroleum fraction showed a significant inhibition of lipoxygenase indicating an anti-inflammatory action (IC₅₀ 29 ± 1 μg/mL). Some of the isolated polymethoxyflavones exhibited strong cytotoxicity against COS7, HeLa and Caco-2 cell lines.     

橙皮苷提取方法的研究

陈皮富含丰富的橙皮苷,是一种抗氧化剂.应用配对比较试验和正交拉丁方试验的方法,以产量和纯度为指标,对3种市售陈皮和碱提酸析和醇提酸析两种提取方法进行比较和筛选.结果表明,从碱石灰和自来水出发的碱提酸析法是合适的橙皮苷工业化提取方法,并找到一种适于橙皮苷工业化的提取方法和来源广、成本低的陈皮原料.     

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.     

Fatty acids,tocopherols, phenolic and antioxidant properties of six citrus fruit species: a comparative study

Citrus fruit is a rich source of bioactive phytochemicals. Information on the fatty acid and tocopherol composition of locally grown citrus fruits in Korea is elusive. This work was aimed to study fatty acid, tocopherols, ascorbic acid, antioxidant potential, and selected phenolics from peel and pulp of six citrus species. The most dominant fatty acid was linoleic acid (15–45%), followed by linolenic, palmitic, and oleic acid. Stearic acid was highly abundant in yuzu peel (14.45%) and pulp (15.88%) compared to the other fruits. Unsaturated fatty acids (54–74%) contributed higher composition than saturated fatty acids (25–46%). Peel exhibited better antioxidant potential and contained higher phytochemicals than pulp. The concentrations of α-tocopherol and γ-tocopherol were ranged from 22.96 (yuzu) to 86.93 (cheonhyeyang) and 38.59 (yuzu) ~83.03 (tangerine) μg/g DW, respectively. J-Redhyeyang peel exhibited highest total flavonoids (4.17?±?0.10 mgQE/g DW) and DPPH radical scavenging activity (6.17?±?0.03 mgTE/g DW). FRAP values were highest (20.05?±?0.64 mgTE/g DW) in yuzu peel, while peel of cheonhyeyang was superior in total phenolic contents. Tangerine (3.02?±?0.05 mg/g DW) and yuzu (7.49?±?0.38 mg/g DW) had higher hesperidin concentrations in pulp and peel, respectively. Naringin was found in an appreciable amount in yuzu pulp (2.04?±?0.09 mg/g DW) and peel (6.30?±?0.19 mg/g DW) but not detected in all other fruit species. Our results indicate that citrus fruit peel is the rich source of antioxidant compounds, which can be used to prepare antioxidant rich food product.     

不同晚熟柑橘中酚类物质的含量检测及分析

采用超高效液相色谱(ultra-performance liquid chromatography,UPLC)对9种晚熟柑橘(沃柑、春见、大雅柑、不知火、默科特、红肉脐橙、伦晚脐橙、红翠2号和塔罗科血橙)中的主要酚类物质(13种类黄酮和7种酚酸)进行测定,分析比较不同品种柑橘果皮、果肉和果汁(柑橘原汁)中酚类物质种类和含量的差异。结果表明:9个柑橘品种的果皮、果肉、果汁中,类黄酮均以橙皮苷为主,酚酸以阿魏酸为主,且果皮中多甲氧基黄酮含量丰富。在9个柑橘品种果汁中,春见的橙皮苷、咖啡酸含量最高。果肉中,沃柑中阿魏酸、咖啡酸、芥子酸含量最高,塔罗科血橙的橙皮苷、对香豆酸含量最高。果皮中,默科特的芸香柚皮苷、川皮苷、阿魏酸含量最高。杂柑果皮中咖啡酸、阿魏酸及川皮苷含量高于橙类。晚熟柑橘含有丰富的酚类物质,并呈现显著的多样性。     

Major Phytochemical Composition of 3 Native Korean Citrus Varieties and Bioactive Activity on V79-4 Cells Induced by Oxidative Stress

ABSTRACT:  Three citrus varieties grown in Korea, namely, Yuza ( Citrus junos  Sieb ex Tanaka), Kjool ( Citrus unshiu  Marcow), and Dangyooja ( Citrus grandis  Osbeck), were evaluated for their dietary fiber, total phenolics, total flavonoids, and total carotenoids as well as individual flavonoids composition. The biochemical characteristics of citrus varieties were examined by the antioxidant capacity (731 to 1221 μmol of Trolox equivalent/g), total phenolic (334 to 411 mg of chlorogenic acid equivalent/100 g), total flavonoid contents (214 to 281 mg of catechin equivalent/100 g), and total carotenoid contents (63 to 84 mg/100 g). Six flavonoids, including naringin, naringenin, hesperidin, hesperetin, neohesperidin, and luteolin, were tentatively identified. Naringin, hesperidin, and neohesperidin were the predominant flavonoids in 3 citrus varieties. Among the citrus varieties studied, Yuza showed higher antioxidant activity, total phenolics, total carotenoids, and lower superoxide radical scavenging assay (SRSA) IC₅₀ values than other varieties. Furthermore, Korean citrus showed higher protective effect on gap-junction intercellular communication (GJIC) as compared to vitamin C and luteolin.     

从柑桔皮中超声提取橙皮苷

从柑桔皮中连续提取色素、果胶和橙皮苷。浸提果胶后的滤渣,在超声波作用下,利用饱和Ca(OH)2碱液浸提盐酸酸析得到橙皮苷。最佳工艺条件为:碱提pH12.5,液料比10:1,超声时间60min,酸析pH4.7,浸提级数3级。另外,考察了重结晶法精制对橙皮苷纯度的影响,平均纯度为94.96%,提高14.72%。红外光谱、高效液相色谱等方法表征实验产品是橙皮苷。     

Hesperetin: A marker of the floral origin of citrus honey

Seventeen flavonoid aglycones were identified in various experimental and commercial citrus honey samples by HPLC analysis. The flavanone hesperetin was detected in all samples. This flavanone was not detected in any of the honey samples, from diverse floral origin (including rosemary, lavender, sunflower, almond, sweet chestnut, white clover, Erisarum, Robinia, Rhododendron, Tilia, Prosopis, Eucalyptus and Calluna honeys) previously investigated. The analysis of the flavonoids present in orange nectar revealed that the flavanone hesperidin (hesperetin-7-rutinoside) was the major flavonoid detected and, therefore, this should be the main source of the hesperetin found in citrus honey. Hesperetin should be produced by hydrolysis of hesperidin by the bee enzymes present in honey. These results suggest that hesperetin could be used as a marker for the botanical origin of citrus honey.