Research on Extraction of Anti-allergic Active Ingredient from Citrus Peels and Its Anti-allergic Activity

[Objective] This study aimed to determine the optimal process conditions for extraction of anti-allergic active ingredient from citrus peels. [Method] Crude anti- allergic active extracts of citrus peels were extracted. Effects of extraction conditions including extraction temperature, ethanol concentration, extraction time and liquid-to-solid ratio on extraction were explored with single-factor experiment, and the extrac- tion process was optimized with orthogonal experiment. [Result] Based on the single- factor experiment and orthogonal experiment, the optimal extraction condition for crude anti-allergic active extracts of citrus peels were determined to be under extraction temperature of 60℃, ethanol concentration of 70%, extraction time of 60 min, liquid-to-solid ratio of 1：25 and twice extraction. [Concluslon] This process condition is of simple operation and high extraction efficiency, which is environment-friendly and suitable for the extraction of crude anti-allergic active extracts from citrus peels. This study provided a new approach and research ideas for the effective and rapid extraction of anti-allergic active ingredient in citrus peels.

Determination of Hesperidin in Citrus Peel by HPLC

[Objective] This study aimed to measure the hesperidin content in citrus peel by high performance liquid chromatography, to provide a scientific basis for quality control and identification. [Method] The hesperidin was extracted with alkaline solution at 70 ~C and pH 6-7, and the purity of hesperidin was determined by HPLC. [Result] The formula for the regression line was Y=466,097Xq3.415 0 （r=0.999 6）, identify- ing the relationship between hesperidin concentration and peak area, and the linear range was 0.2-1.4μg. The hesperidin solution was stable within 24 h at room temperature. The average recovery rate of hesperidin was 98.41%. [Conclusion] The HPLC method is rapid, simple, and with good linear relationship, can be used for routine analysis of hesperidin.

Human plasma protein binding of water soluble flavonoids extracted from citrus peels

The human plasma protein binding of water soluble flavonoids in the peels of five spices of citrus fruits was studied by ultrafiltration combined with HPLC.The flavonoids were extracted separately by hot and cold water,and higher total flavonoid contents were detected in the former extracts than the latter ones.All the extracts show significant scavenging abilities to both ABTS and DPPH free radicals,which indicates the health benefits of the water extracts of citrus fruits peels.For DPPH radical,the IC50values of hot extract follow as Navel orange(NO)≈Mandarin orange(MO)< Lemon(LE)< Lo tangerine(LO)< Pomelo(PO),while the rank is NO< PO<LE≈MO<LO for ABTS radical.The HPLC results reveal that the kinds and contents of the flavonoids detected in the extracts are different among the species.MO extract has the most neohesperidin dihydrochalcone of 118.76 μmol/L and quercetrin of 211.81 μmol/L of which are much more than the rest extracts.Pomelo extract has the most plentiful flavonoids of naringin with a concentration of 303.28 μmol/L.The high contents of myricetrin and dihydromyricetin which both are potent free radical scavengers may explain the highest free radical scavenging activity of the NO extract.The plasma binding rates decrease with the increasing concentrations of flavonoids,and the flavonoids having plenty hydroxyl groups on both A ring and B ring of the molecular skeleton have relative higher plasma binding rates.In addition,the plasma binding rates of flavonoids with saturated C3-C4 bond decrease significantly with the increasing concentrations.

Efficacious anti-cancer property of flavonoids from citrus peels

Cancer is one of the two leading human fatal diseases.Drug development for cancer intervention has progressed well in past decades yet existing drugs face many limitations in applications and effectiveness and are often associated with serious of side effects,which can further deteriorate the patients’quality of life.Recent development of natural product based and therapeutically sound anti-cancer agents have gained popularity in the field of functional foods,in which a few have demonstrated efficacy and minimal toxicity toward the prevention and treatment of carcinogenesis.With multiple active molecular components,citrus peels and derived extracts have demonstrated potent efficacious properties against various cancers due in large part to the rich content of flavonoids present in citrus peels.This review summarizes the results of currently available data regarding the in vivo anti-cancer activity of citrus peel flavonoids,and identifies opportunities for subsequent human clinical trials to assess preventive and therapeutic effects in the near future.

Formulated citrus peel extract gold lotion improves cognitive and functional recovery from traumatic brain injury(TBI)in rats

Traumatic brain injury(TBI)is closely related to neuro-inflammatory response and causes a complex pathological process and serious consequence.In this study,we explored whether the formatted citrus peel extract gold lotion(GL)could have a therapeutic effect on rats suffered TBI.TBI rat model was prepared by the electric Cortical Contusion Impactor(eCCI)device.Prevention against TBI by GL was assessed by the behavioral,organizational of rats and the molecular markers in the brain injury area.The results showed that GL could reduce nerve damage and neurological dysfunction.The mechanisms probably due to that GL could not only reduce the associated damage markers including GFAP,OX-42,TNF-α,COX-2,NF-κB and TLR4 but also promote the expression of VEGF which is responsible for protecting neuronal cells in the injury area.These findings revealed that GL could be a promising preparation for TBI prevention.

Formulation and Optimization of Constituent in Legumes-Based Milk Chocolate Fortified with Citrus Peel Powder

The present study was employed to optimize the process for development of legumes-milk based chocolate using peanut (PN) and yellow-pea (YP) milk using Response surface methodology (RSM). Different combinations of legumes-milk chocolate at various ratios of PN:YP milk with fixed concentration of other regular ingredients were prepared, and the finest combination (1:1) was selected on the basis of their sensory and nutritional properties. PN and YP milk, Jaggery (JG), Butter (BT), and Citrus Peel Powder (CPP) served as independent variables while the dependent variables were allocated to the regression equation to determine folic acid (R2 = 93.15) along with protein content (R2 = 93.11) and vitamin C (R2 = 90.57). The nutritional parameters such as Folic acid, Protein and Vitamin C content were found to be optimum in the milk chocolate. The optimized concentrations of PNM, YPM, JG, BT and CPP were found to be 5.0 ml, 5.5 ml, 18.1 g, 2.9 g and 0.53 g respectively. Addition of JG had an interactive effect on folic acid in YP milk (p < 0.05) and CPP shows a significant. Enhancement of vitamin C was perceived in legume based milk chocolate than the control chocolate due to supplement of CPP.

Assessing the bioactivity,cytotoxicity,and rheological properties of pectin recovered from citrus peels

The recovery of pectin from citrus peel waste for application in the food,nutraceutical,pharmaceutical,and medical industries has attracted much interest.It has been postulated that the presence of polyphenols,sometimes regarded as an impurity,in extracted pectin might confer enhanced biological property.This research study aimed at investigating the physicochemical and biological properties(antioxidant,enzyme inhibitory,and cytotoxic properties)of pectin extracted from the alcohol insoluble residues of the peel waste of grapefruit,lime,and lemon as well as the pectin recovered from untreated peel waste.The rheological behavior of the pectin samples was also evaluated.High methoxyl pectin with galacturonic acid content greater that 60%was recovered from the different raw materials.Pectin obtained from untreated material possessed high phenolic content(19.83-5.94 mg gallic acid equivalent/g sample)and showed higher antioxidant activity.Inhibition against pancreatic cholesterol esterase,pancreatic lipase,andα-glucosidase was observed and pectin enriched with polyphenol showed higher activity.Addition of pectin samples to healthy Vero cells enhanced proliferation and did not show cytotoxic effect.Pectin solutions extracted from untreated lime and untreated grapefruit presented highest viscosity and elastic modulus,which were comparable to those obtained for commercial high-methoxyl pectin.Pectin extracted from lime,lemon,and grapefruit peels possessed interesting biological and viscoelastic properties,which can be exploited for development of novel bioproducts.

Protective effects of a polymethoxy flavonoids-rich Citrus aurantium peel extract on liver fibrosis induced by bile duct ligation in mice

Objective: To evaluate the possible protective effect of Citrus aurantium peel extract(CAE) against apoptosis in cholestatic liver fibrosis induced by bile duct ligation in mice. Methods: Male ICR mice were divided to 5 groups: 1) Control group(Sham-operated mice), 2) Cholestatic liver injury group induced by bile duct ligation(BDL), 3) BDL mice treated with silymarin(200 mg/kg) for 4 weeks, 4) BDL mice treated with 50 mg/kg CAE for 4 weeks, 5) BDL mice treated with 200 mg/kg CAE for 4 weeks. Mice were sacrificed and liver fibrosis was evaluated by serum and hepatic tissue biochemistry tests and liver histopathological examination. Effects of CAE on inflammation and apoptosis gene regulation were investigated through real-time PCR. CAE effect on lipid metabolism related signaling was determined by western blot analysis. Results: In BDL mice, administration of CAE for 4 weeks markedly attenuated liver fibrosis based on histopathological alteration. Serum and hepatic tissue biochemistry results revealed that CAE(50 and 200 mg/kg) decreased the levels of alanine transaminase, aspartate transaminase, gamma glutamyl transferase, total bilirubin, nitric oxide, and thiobarbituric acid reactive substances. Real-time PCR and western blot analysis showed that CAE regulated inflammation, apoptosis, and lipid metabolism factors increased by BDL. Interleukin family, tumor necrosis factor α, and related apoptosis factors m RNA levels were increased by BDL treatment. However, these increases were suppressed by CAE administration. In addition, CAE effectively increased phosphorylation of AMPactivated protein kinase, nuclear factor E2-related factor 2, and related cytoprotective proteins. Conclusions: CAE can efficiently regulate BDL-induced liver injury with antioxidant, antiinflammatory, and antiapoptotic activities.

Study on Infrared Spectra of Citrus grandis Peels from Guangxi Zhuang Autonomous Region

[ Objective ] This study aimed to establish the infrared （IR） spectra fingerprints for peels of Citrus grandis from Guangxi Zhuang Autonomous Region. [ Method ] The fingerprint was established by fourier transform infrared spectroscopy （ FT-IR）, and then were analyzed by Omic software. [ Result ] IR spectra fin- gerprint of C. grand/s were successfully established. There were six common peaks in the characteristic spectra of 12 samples. [ Conclusion] The specific, rapid and stable method can be applied to quality control of C. grandis.

柑桔果皮中多甲氧基黄酮的LC-APCI-MS定性分析

黄酮化合物是具有多种生物活性的物质,广泛存在于自然界中.单纯为甲氧基取代的多甲氧基黄酮PMF是芸香科多种桔类所特有的,特别是在其果皮中有较高的含量.近年来有关桔皮PMF抗癌的研究取得了很大进展.PMF作为桔皮中的抗癌活性成分,对多种癌细胞有较强的抑制作用,受到国内外的广泛关注.……

Identification and fermentation optimization of protopectinase-overproducing strain Aspergillus niger CD-01 for pectin production

In order to solve the citrus peel resource waste problem protopectinase-overproducing strain CD-01 for pectin production and minimize the drawbacks of chemical extraction of pectin, a was isolated from a pit soil dumped with perished orange in Changde City, Hunan Province of China. The strain CD-01 had the same morphology and 28S rRNA gene sequence （FJ184995） as that of Aspergillus niger （ATCC 64028）. It was thus identified and named as Aspergillus niger CD-01. The fermentation condition was optimized based on L9（34） orthogonal experimental design and the variances analyses. The results show that the optimal condition for producing pectin is as follows： time 36 h, temperature 35 ℃, pH 5, and urea as the nitrogen source. Under this condition, the pectin yield can reach up to 24.5%. This shows a great potential of Aspergillus niger CD-01 in pectin extraction from citrus.

5-Acetyl-6,7,8,4′-tetramethylnortangeretin induces apoptosis in multiple myeloma U266 cells

Multiple myeloma(MM)is one of the most common hematological malignancies and to date,it remains an incurable disease.In this study,we evaluated the inhibitory effect of 5-acetyloxy-6,7,8,4′-tetramethoxyflavone(5-ATMF),a compound from aged citrus peel extracts,on the MM U266 cell line.We found that the compound inhibited cell growth and induced cell apoptosis in multiple apoptotic assays.The apoptotic proteins caspase-3,caspase-9,and PARP(poly ADP-ribose polymerase)were cleaved when cells were treated with 5-ATMF.Along with the apoptosis process,the anti-apoptotic protein,Bcl-2(B-cell lymphoma-2),was significantly downregulated and the pro-apoptotic protein,Bax(Bcl-2 associated X protein),upregulated along with the release of cytochrome C(Cyt c)and the reduction of mitochondrial membrane potential(MMP).Notably,we found that the phosphorylation of Bad(Bcl-2/Bcl-XL-associated death promoter)was decreased but Bad remained unchanged in this process.On adding 5-ATMF to U266 cells,we observed that 5-ATMF repressed the phosphorylation of Akt(PKB Protein Kinase B PKB),thereby increasing the release of Cyt c and inhibiting the phosphorylation of Bad.These effects were enhanced by Ly294002,an inhibitor of Akt.These results suggested that 5-ATMF exerts a pro-apoptotic effect on U266 cells,possibly associated with the mitochondrial apoptotic pathway induced by the PI3K/Akt/Bad pathway.

New approach of medicinal herbs and sulfasalazine mixture on ulcerative colitis induced by dextran sodium sulfate

BACKGROUND Sulfasalazine has been used as a standard-of-care in ulcerative colitis for decades,however,it results in severe adverse symptoms,such as hepatotoxicity,blood disorders,male infertility,and hypospermia.Accordingly,the new treatment strategy has to enhance pharmacological efficacy and stimultaneously minimize side effects.AIM To compare the anti-inflammatory action of sulfasalazine alone or in combination with herbal medicine for ulcerative colitis in a dextran sodium sulfate(DSS)-induced colitis mouse model.METHODS To induce ulcerative colitis,mice received 5%DSS in drinking water for 7 d.Animals were divided into five groups(n=9 each)for use as normal(non-DSS),DSS controls,DSS+sulfasalazine(30 mg/kg)-treatment experimentals,DSS+sulfasalazine(60 mg/kg)-treatment experimentals,DSS+sulfasalazine(30 mg/kg)+Citrus unshiu peel and Bupleuri radix mixture(30 mg/kg)(SCPB)-treatment experimentals.RESULTS The SCPB treatment showed an outstanding effectiveness in counteracting the ulcerative colitis,as evidenced by reduction in body weight,improvement in crypt morphology,increase in antioxidant defenses,down-regulation of proinflammatory proteins and cytokines,and inhibition of proteins related to apoptosis.CONCLUSIONSCPB may represent a promising alternative therapeutic against ulcerative colitis,without inducing adverse effects.