• Korean Journal of Pharmacognosy
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• v.40 no.4
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• pp.370-375
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• 2009

Overproduction of peroxynitrite ($ONOO^-$) causes a variety of disease such as atherosclerosis, hypercholesterolemia, diabetes mellitus or obesity. Peroxynitrite scavenging activities and HPLC analysis on the five Oriental medicinal drugs belonging to the genus Citrus, Aurantium or Poncirus (Rutaceae family) and HPLC analysis were taken to evaluate flavanone glycosides with peroxynitrite scavenging activity. The $IC_{50}s$ of the five crude drugs were shown as follows: Aurantii nobilis Pericarpium (Jinpi, 18.3 ${\mu}g$/ml), Citrii unshiu Pericarpium (Chungpi, 7.50${\mu}g$/ml), Citrii unshiu Semen (Gyulhaek, >50.0${\mu}g$/ml), Aurantii Fructus (Jigak, 18.3${\mu}g$/ml), and Poncirii Fructus (Jisil, >50.0${\mu}g$/ml) where Korean crude drug's names are noted in the parenthesis. Peroxynitrite scavenging effect of flavanones or their glycosides usually contained in Citrus species were observed as follows: hesperetin (1.89 ${\mu}g$/ml), naringenin (7.77 ${\mu}g$/ml), hesperidin (8.44 ${\mu}g$/ml), poncirin (>50.0 ${\mu}g$/ml)and ponciretin(>50.0 ${\mu}g$/ml). The activities of naringin and poncirin with ${\alpha}$-L-rhamnopyranosyl($1{\rightarrow}2$)-${\beta}$-D-glucopyranosyl moiety were weak. HPLC analytical data revealed that Jinpi (the peels of mature fruits of Citrus unshiu) and Chungpi (the peels of immature fruits of C. unshiu) had high quantities of hesperidin as the value of 142.1${\pm}$0.21 and 104.51${\pm}$1.10 mg/g dried weight, respectively. Poncirin was clearly detected in only Jisil and naringenin and naringin were not observed on the HPLC chromatogram of the five crude drugs.

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.442-448
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• 2014

The major objective of this study was to investigate the usability as cultivar for processing of new Citrus hybrid 'Hamilgam'. We investigated various antioxidant activities, such as DPPH radical, ABTS radical, Hydroxyl radical and Superoxide anion radical effect, along with total polyphenol content and total flavonoid content of Citrus hybrid 'Hamilgam', C. unshiu, C. natsudaidai. The total polyphenol content and total flavonoid content were the highest in the Hamilgam. The major flavonoids of Hamilgam and Natsudaidai were naringin and neohesperidin, and those of Unshu were narirutin and hesperidin, as determined by HPLC. Especially, the neohesperidin content of Hamilgam showed much higher (100 times over) than that of Unshu. However, the aglycone form was not found. DPPH and ABTS radical scavenging activities were the highest in Hamilgam peels. Hydroxyl radical scavenging activity was high in the order of Unshu, Hamilgam and Natsudaidai. Superoxide anion radical scavenging of 3 cultivars peels displayed low activities compared to DPPH and ABTS radical scavenging. Based on these results, Hamilgam peel extracts possess antioxidant activities and may thus serve as potential sources of functional food, cosmetic products, etc.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.3
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• pp.331-337
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• 2015

Citrus and its peels, which are by-products from juice and/or jam processing, have long been used in Asian folk medicine. Citrus peels show an abundant variety of flavanones, and these flavanones have glycone and aglycone forms. Aglycones are more potent than glycones with a variety of physiological functions since aglycone absorption is more efficient than glycones. Bioconversion with cytolase converted narirutin and naringin into naringenin and hesperidin into hesperetin. Therefore, this study aimed to investigate the anti-oxidant and anti-inflammatory effects of bioconversion of Citrus unshiu (CU) peel extracts with cytolase (CU-C) in RAW264.7 cells. HPLC chromatograms showed that CU and CU-C had 23.42% and 29.39% total flavonoids, respectively. There was substantial bioconversion of narirutin to naringenin and of hesperidin to hesperetin. All citrus peel extracts showed DPPH scavenging activities in a dose-dependent manner, and CU-C was more potent than intact CU. RAW264.7 cells were pre-treated with $0{\sim}500{\mu}g/mL$ of citrus peel extracts for 4 h and then stimulated by $1{\mu}g/mL$ of lipopolysaccharide (LPS) for 8 h. All citrus peel extracts showed decreased mRNA levels and protein expression of LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner. Especially, CU-C markedly inhibited mRNA and protein expression of iNOS and COX-2 compared to intact citrus peel extracts. All citrus peel extracts showed decreased NO production by iNOS activity. This result suggests that bioconversion of citrus peel extracts with cytolase may provide potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by boosting the anti-inflammatory effects of citrus peels.

• Korean Journal of Pharmacognosy
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• v.49 no.2
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• pp.113-123
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• 2018

The application of 112 samples of Citrus fruit peel, which is commonly cultivated in Korea as crude drug, has been limited due to the absence of criteria concerning chemical constituents and application of itself. In the context, this study is on the analysis of 11 flavonoid compounds. As a result, the flavonoid pattern could be classified into 5 groups. Furthermore, by utilizing this HPLC data, this study could provide a basis for fruit peel applications of not only Citrus unshiu, but also its cultivars, hybrids and some of Citrus plants which has been cultivated in Jeju Island for a long time. Moondan could be used for the same purposes as Chinese Huajuhong, and Poncirus trifoliata of Korean Jisil origin are effective enough, in terms of ingredients and function, compared with C. aurantium. Based on the content and composition of the PMF region in this study, C. unshiu and C. reticulata, as Tanaka System shows, should be considered as different hortcultural speicies. Also, Jikak, which is known as 'sour orange' until now, is confirmed as Chosen-daidai, C. tenuissima, that was reported by Tanaka in 1970, and this C. tenuissima could be used for the same purposes as sour orange.

• Natural Product Sciences
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• v.21 no.2
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• pp.141-145
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• 2015

Hesperidin, which is the most abundant flavonoid of Citrus unshiu (Rutaceae), has been reported to possess diverse activities and widely used as functional foods and cosmetics. For the development of functional products, extraction procedure is indispensable. Extraction conditions affect the composition of extract as well as its biological activity. Therefore, we tried to optimize extraction conditions such as extraction solvent, extraction time and extraction temperature for maximum yield of hesperidin using response surface methodology with threelevel-three-factor Box-Behnken design (BBD). Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as ethanol concentration, 59.0%; temperature $71.5^{\circ}C$ and extraction time, 12.4 h. The hesperidin yield under the optimal condition was found to be $287.8{\mu}g$ per 5 mg extract, which was well matched with the predicted value of 290.5 μg. These results provides optimized extraction condition for hesperidin and might be useful for the development of hesperidin as functional products like health supplements, cosmetics and medicinal products.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.9
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• pp.1476-1485
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• 2004

Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

• Journal of Life Science
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• v.24 no.3
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• pp.260-265
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• 2014

Dried Citrus unshiu peels (Aurantii Nobilis Pericarpium; ANP) are used as a traditional folk medicine for the treatment of gastrointestinal (GI) motility disorders in East Asia, including Korea. In the present study, an ethanolic extract of ANP (ANP-E) exhibited no significant toxicity in mice, even at an oral dose of 5 g/kg. The effects of ANP-E on GI motor function were investigated by measuring the intestinal transit rate (ITR) of Evans blue in normal mice and mice with experimental GI motility dysfunction (i.e., peritoneal irritation by acetic acid; PIA). In normal mice, ANP-E significantly increased the ITR in a dose-dependent manner. The ITR in the PIA mice was significantly retarded compared to that in the normal mice. However, ANP-E significantly inhibited this retardation in a dose-dependent manner. Furthermore, in all the models, the potency of ANP-E appeared to be same or higher than that of cisapride, which was used predominantly for the treatment of various GI motility disorders in humans in the 1900s but was removed from the market in 2000 due to fatal side effects. The results suggest that ANP-E has potential as a new prokinetic agent that could be used as a substitute for cisapride.

• Korean Journal of Food Science and Technology
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• v.45 no.5
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• pp.598-604
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• 2013

This study was designed to evaluate the usability of the following citrus peel extracts (CPEs): Citrus sinensis (orange), C. unshiu (mandarin orange), C. limon (lemon), and C. paradise (grapefruit) as natural antibrowning agents. Overall, 0.1% of the CPEs were effective in reducing the browning of apple slices. The appearance of apple slices dipped in C. limon peel extracts (LPE) were found to be excellent and their Hunter L and ${\Delta}E$ values were similar to the values obtained when apple slices were dipped in ascorbic acid. The pH values of the different CPEs were not significantly different from each other, but were higher than that of ascorbic acid. LPE was found to possess the highest total phenolic content, flavonoid content, DPPH radical scavenging activity, and copper ($Cu^{2+}$) chelating activity. All these results suggest that citrus peel extracts, especially lemon peel extract, can be used as natural antibrowning agents.

• Korean Journal of Food Science and Technology
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• v.31 no.5
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• pp.1178-1183
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• 1999

Essential oils were isolated from the peel of mandarin (Citrus unshiu Marc. Var. Okitsu) cultivated in Cheju Island. Among three different isolation methods investigated, simultaneous distillation gave higher yield 1.14%(w/w) than solvent extraction or cold pressing. The densities of essential oils were between 0.8409 and 0.8530, which showed no significant effect of the isolation methods. Some variations in the number of constituents detected and the content of d-limonene in the peel oils were observed depending upon the isolation methods. The citrus fruit was collected seasonally from the field and used as a sample for analysis. The peel thickness and the peel content of citrus fruit were around 2 mm and about 17% on wet weight basis, respectively, in harvest season. The maximum yield of essential oils was obtained in September from citrus peels just prior to ripeness and thereafter the yield had a tendency of decrease but remained over 1%(w/w). The relative content of d-limonene, the major constituent of citrus peel oils. increased gradually with ripening up to 68.69%(relative peak area, RPA). The other minor constituents were ${\gamma}-terpinene,\;{\beta}-elemene$, farnesene, hexadecanoic acid, ${\alpha}-pinene,\; {\beta}-myrcene$ and linalool in the decreasing order of their own contents of 7.75, 2.96, 2.29, 1.76, 1.63, 1.56 and 1.46%(RPA).

• Journal of Applied Biological Chemistry
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• v.50 no.3
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• pp.148-154
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• 2007

Volatile components of the essential oils of Satsuma mandarin (C. unshiu), Dangyuza (C. grandis), Yuza (C. junos), Byungkyul (C. playtymamma), Jinkyul (C. sunki), and Hakyul (C. natsudaidai) grown in Jeju Island were isolated from the fruit peels by hydro distillation and determined by GC-MS. GC-MS analysis identified 58 compounds, with main components being d-limonene $(64.01{\sim}79.34%),\;{\beta}-myrcene\;(3.01{\sim}26.53%),\;{\gamma}-terpinene\;(0.11{\sim}12.88%),\;{\beta}-pinene\;(0.78{\sim}4.74%),\;and\;{\alpha}-pinene\;(1.01{\sim}2.55%)$. Differences in compositions and contents of the essential oils were observed among citrus varieties. Effects of citrus oils on growth inhibitions of Escherchia coli, Staphyllococcus epidermidis, and Candida albicans were investigated using disc diffusion assay and minimal inhibitory concentration (MIC) assay. The essential oils inhibited growths of the test organisms, exhibiting higher levels of activity against Gram-positive S. epidermidis (MIC values $0.04{\sim}0.17mg/mL$), whereas Gram-negative E. coli was moderately resistant (MIC values $1.66{\sim}20.30mg/mL$). MIC of citrus essential oils ranged from $0.82{\sim}23.69mg/mL$ against C. albicans. The essential oils obtained from C. sunki, C. grandis, and C. playtymamma showed the highest antimicrobial activities against S. epidermidis and C. albicans, indicating their potential as natural antimicrobial agents.