不同萃取溶劑對餘甘子抗氧化性、澱粉水解酶抑制活性及糖化最終產物形成之影響

Effects of Different Extraction Solvents on Antioxidant Activities, Starch Hydrolase Inhibitory Activities and Advanced Glycation End Products Formation of Phyllanthus emblica Linn

10.6578/TJACFS.202012_58(6).0004

江文德(Wen-Dee Chiang)；劉上詠(Shang-Yung Liu)；高馥君(Fuh-Juin Kao)

余甘子 ； 可水解單寧 ； 抗氧化活性 ； 澱粉水解酶 ； 糖化最終產物 ； Phyllanthus emblica Linn. ； Hydrolyzable tannin ； Antioxidant activity ； Starch hydrolase ； Advanced glycation end products

臺灣農業化學與食品科學

58卷6期（2020 / 12 / 01）

163 - 175

繁體中文

本研究在比較沸水、50%甲醇、50%乙醇、95%甲醇、95%乙醇與乙酸乙酯等六種餘甘子萃取物對其自由基清除活性（DPPH、超氧陰離子、NO）、脂肪過氧化抑制活性、澱粉水解酶（α-澱粉酶、α-葡萄糖苷酶）抑制活性及糖化最終產物（AGEs）形成抑制活性之影響，另外也分析各萃取物中活性成份含量。結果發現，以50%甲醇及50%乙醇萃取物的收率、總類黃酮含量與可水解單寧含量最高，而95%甲醇及95%乙醇萃取物的總酚含量、NO清除活性則是最高。所有甲、乙醇萃取物在DPPH自由基清除活性、脂肪過氧化抑制活性、α-葡萄糖苷酶抑制活性與AGEs抑制活性上，皆顯著高於沸水及乙酸乙酯萃取物。HPLC分析結果顯示，沒食子酸苷在六種萃取物中的含量皆是最高，尤以乙酸乙酯萃取物中的含量最豐富。除此之外，所有醇類萃取物中，以50%甲醇及50%乙醇萃取物，具高抑制AGEs形成活性、高α-葡萄糖苷酶抑制活性與較溫和的α-澱粉酶抑制活性，上述的結果指出50%甲醇及50%乙醇萃取物具有潛力開發為營養保健品，用於管理第二型糖尿病。

The study was to compared the effects of different solvents (boiling water, 50% methanol, 50% ethanol, 95% methanol, 95% ethanol and ethyl acetate) extraction of Phyllanthus emblica Linn. on their scavenging activity of DPPH radical, superoxide radical and NO as well as inhibitory activity of lipid peroxidation, starch hydrolase (α-amylase, α-glucosidase) and advanced glycation end products (AGEs) formation. Furthermore, active compounds of each extract were also determined. The results showed that extracts of 50% methanol and 50% ethanol had highest yield, total flavonoid content and hydrolyzable tannin content, while extracts of 95% methanol and 95% ethanol exhibited the maximum total phenolics content and NO scavenging activities. All the extracts of methanol/ethanol exhibited superior DPPH radical scavenging activities as well as inhibitory activities of lipid peroxidation, α-glucosidase and AGEs formation than extracts of boiling water and ethyl acetate. Based on HPLC analysis, all the extracts, especially ethyl acetate extract, were rich in β-glucogallin. In addition, all methanol/ethanol extracts, especially extracts of 50% methanol and 50% ethanol, showed strong inhibitory activities of AGEs formation and α-glucosidase but mild inhibitory activity of α-amylase. The results indicate that extracts of 50% methanol and 50% ethanol have the potential to be used as nutraceutical in the management of type 2 diabetes.

1. Arora, S.,Kaur, K.,Kaur, S.(2003).Indian medicinal plants as a reservoir of protective phytochemicals.Teratogen. Carcin. Mut.,1,295-300.
2. Baliga, M. S.,Dsouza, J. J.(2011).Amla (Emblica officinalisGaertn), a wonder berry in the treatment and prevention of cancer.Eur. J. Cancer Prev.,20,225-239.
3. Bandyopadhyay, S. K.,Pakrashi, S. C.,Pakrashi, A.(2000).The role of antioxidant activity of Phyllanthus emblica fruits on prevention from Indomethacin induced gastric ulcer.J. Ethnopharmacol.,70,171-176.
4. Chang, K. C.,Laffin, B.,Ponder, J.,Énzsöly, A.,Németh, J.,LaBarbera, D. V.,Petrash, J. M.(2013).Beta-glucogallin reduces the expression of lipopolysaccharide-induced inflammatory markers by inhibition of aldose reductase in murine macrophages and ocular tissues.Chem.-Biol. Interact.,202,283-287.
5. Charoenteeraboon, J.,Ngamkitidechakul, C.,Soonthornchareonnon, N.,Jaijoy, K.,Sireeratawong, S.(2010).Antioxidant activities of the standardized water extract from fruit of Phyllanthus emblica Linn.Songklanakarin J. Sci. Technol.,32,599-604.
6. Chen, J.,Li, L.,Zhou, X.,Li, B.,Zhang, X.,Hui, R.(2018).Structural characterization and α-glucosidase inhibitory activity of polysaccharides extracted from Chinese traditional medicine Huidouba.Int. J. Biol. Macromol.,117,815-819.
7. Dasaroju, S.,Gottumukkala, K. M.(2014).Current trends in the research of Emblica officinalis (Amla): A pharmacological perspective.Int J Pharm Sci Rev Res.,24,150-159.
8. Elendran, S.,Wang, L. W.,Prankerd, R.,Palanisamy, U. D.(2015).The physicochemical properties of geraniin, a potential antihyperglycemic agent.Pharm. Biol.,53,1719-1726.
9. Felhi, S.,Daoud, A.,Hajlaoui, H.,Mnafgui, K.,Gharsallah, N.,Kadri, A.(2017).Solvent extraction effects on phytochemical constituents profiles, 2 antioxidant and antimicrobial activities and functional group analysis of Ecballium elaterium seeds and peels fruits.Food Sci. Technol.,37,483-492.
10. Fernandes, L.,Prereira, J. A.,López-Cortés, I.,Salazar, D. M.,Ramalhosa, E. C.(2015).Physicochemical changes and antioxidant activity of juice, skin, pellicle and seed of pomegranate (cv. Mollar de Elche) at different stages of ripening.Food Technol. Biotechnol.,53,397-406.
11. Fitriansyah, S. N.,Aulifa, D. L.,Febriani, Y.,Sapitri, E.(2018).Correlation of total phenolic, flavonoid and carotenoid content of Phyllanthus emblica extract from Bandung with DPPH scavenging activities.Pharmacogn. Mag.,10,447-452.
12. Gambari, R.,Borgatti, M.,Lampronti, I.,Fabbri, E.,Brognara, E.,Bianchi, N.,Piccagli, L.,Yuen, M. C.,Kan, C. W.,Hau, D. K.,Fong, W. F.,Wong, W. Y.,Wong, R. S.,Chui, C. H.(2012).Corilagin is a potent inhibitor of NF kappa B activity and downregulates TNF-alpha induced expression of IL-8 gene in cystic fibrosis IB3-1 cells.Int. Immunopharmacol.,13,308-315.
13. Ghosal, S.,Tripathi, V. K.,Chauhan, S.(1996).Active constituent of Emblica officinalis: part 1.the chemistry and antioxidant effects of two new hydrolysable tannins, emblicanin A and B.Indian J. Chem.,35B,941-948.
14. Hayouni, E. A.,Abedrabba, M.,Bouix, M.,Hamdi, M.(2007).The effects of solvents and extraction method on the phenolic contents and biological activities in vitro of Tunisian Quercus coccifera L. and Juniperus phoenicea L. fruit extracts.Food Chem.,105,1126-1134.
15. Hsu, C. Y.,Chan, Y. P.,Chang, J.(2007).Antioxidant activity of extract from Polygonum cuspidatum.Biol Res,40,13-21.
16. Huang, C. Z.,Tung, Y. T.,Hsia, S. M.,Wu, C. H.,Yen, G. C.(2017).The hepatoprotective effect of Phyllanthus emblica L. fruit on high fat diet-induced non-alcoholic fatty liver disease (NAFLD) in SD rats.Funct.,8,842-850.
17. International Diabetes Foundation. IDF diabetes atlas, 9th edition 2019. 取自http://www.diabetesatlas.org/
18. Jacob, A.,Pandey, M.,Kapoor, S.,Saroja, R.(1988).Effect of the Indian gooseberry (amla) on serum cholesterol levels in men aged 35-55 years.Eur. J. Clin. Nutr.,42,939-944.
19. Julius, A.,Hopper, W.(2017).Inhibition of advanced glycation end-product formation by quercetin and catechin: an alternative therapy for treating diabetic complications.Asian J. Pharm. Clin. Res.,10,173-176.
20. Khan, K. H.(2009).Roles of Emblica officinalis in medicine – A review.J. Bot.,2,218-228.
21. Khan, M.,Liu, H.,Wang, J.,Sun, B.(2020).Inhibitory effect of phenolic compounds and plant extracts on the formation of advance glycation end products: A comprehensive review.Food Res. Int.,130,1-14.
22. Kim, H. K.,Cheon, B. S.,Kim, Y. H.,Kim, S. Y.,Kim, H. P.(1999).Effects of naturally occurring flavonoids on nitric oxide production in the macrophage cell line Raw 264.7 and their structure–activity relationships.Biochem. Pharmacol.,58,759-765.
23. Klika, K. D.,Saleem, A.,Sinkkonen, J.,Kähkönen, M.,Loponen, J.,Tähtinen, P.,Pihlaja, K.(2004).The structural and conformational analyses and antioxidant activities of chebulinic acid and its thrice-hydrolyzed derivative, 2, 4-chebuloyl-β-D-glucopyranoside, isolated from the fruit of Terminalia Chebula.Arkivoc,7,83-105.
24. Liu, X. L.,Cui, C.,Zhao, M. M.,Wang, J. S.,Luo, W.,Yang, B.,Jiang, Y.(2008).Identification of phenolics in the fruit of emblica and (Phyllanthus emblica L.) their antioxidant activities.Food Chem.,109,909-915.
25. Liu, X.,Zhao, M.,Wang, J.,Yang, B.,Jiang, Y.(2008).Antioxidant activity of methanolic extract of emblica fruit (Phyllanthus emblica L.) from six regions in China.J. Food Compos. Anal.,21,219-228.
26. Luo, W.,Zhao, M.,Yang, B.,Ren, J.,Shen, G.,Rao, G.(2011).Antioxidant and antiproliferative capacities of phenolics purified from Phyllanthus emblica L. fruit.Food Chem.,126,277-282.
27. Majeed, M.,Bhat, B.,Anand, T. S.(2010).Inhibition of UV induced adversaries by β-glucogallin from Amla (Emblica officinalis Gaertn.) fruits.Indian J. Nat. Prod. Resour.,1,462-465.
28. Matsuura, N.,Aradate, T.,Sasaki, C.,Kojima, H.,Ohara, M.,Hasegawa, J.,Ubukata, M.(2002).Screening system for the Maillard reaction inhibitor from natural product extracts.J. Health Sci.,48,520-526.
29. Mogale, M.,Lebelo, L. S.,Thovhogi, N.,de Freitas, A. N.,Shai, L. J.(2011).α-amylase and α-glucosidase inhibitory effects of Sclerocarya birrea [(A. Rich.) Hochst.] subspecies caffra (Sond) Kokwaro (Anacardiaceae) stem-bark extracts.Afr. J. Biotechnol.,10,15033-15039.
30. Nampoothiri, S. V.,Prathapan, A.,Cherian, O. L.,Raghu, K. G.,Venugopalan, V. V.,Sundaresan, A.(2011).In vitro antioxidant and inhibitory potential of Terminalia bellerica and Emblica officinalis fruits against LDL oxidation and key enzymes linked to type 2 diabetes.Food Chem. Toxicol.,49,125-131.
31. Nguyen, V. T.,Bowyer, M. C.,Vuong, Q. V.,Altena, I. A. V.,Scarlett, C. J.(2015).Phytochemicals and antioxidant capacity of Xao tam phan (Paramignya trimera) root as affected by various solvents and extraction methods.Ind. Crops Prod.,67,192-200.
32. Nonaka, G. I.,Nishioka, I.,Nishizawa, M.,Yamagishi, T.,Kashiwada, Y.,Dutschman, G. E.,Bonder, A.,Kilkuskie, R. E.,Cheng, Y. C.,Lee, K. H.(1990).Anti-AIDS agents, 2: Inhibitory effect of tannins on HIV reverse transcriptase and HIV replication in H9 lymphocyte cells.J. Nat. Prod.,53,587-595.
33. Palanisam, U. D.,Teng, L. L.,Manaharan, T.,Appleton, D.(2011).Rapid isolation of geraniin from Nephelium lappaceum rind waste and its anti-hyperglycemic activity.Food Chem.,127,21-27.
34. Pietta, P. G.(2000).Flavonoids as antioxidants.J. Nat. Prod.,63,1035-1042.
35. Poongunran, J.,Perera, H. K. I.,Fernando, W. I. T.,Jayasinghe, L.,Sivakanesan, R.(2015).α-Glucosidase and α-amylase inhibitory activities of nine Sri Lankan antidiabetic plants.J. Pharm. Res. Int.,7,365-374.
36. Raman, S.,Akbor, M. M.,Howlader, A.,Jabbar, A.(2009).Antimicrobial and cytotoxic activity of the alkaloids of amlaki (Emblica officinalis).Pak. J. Biol. Sci.,12,1152-1155.
37. Ramani, R.,Anisetti, R. N.,Boddupalli, B. M.,Malothu, N.,Arumugam, B. S.(2013).Antiparkinson’s and free radical scavenging study of ethyl acetate fraction of ethanolic extract of Leucas lanata.Drug invent. today,5,251-255.
38. Ramkissoon, J. S.,Mahomoodally, M. F.,Ahmed, N.,Subratty, A. H.(2013).Antioxidant and anti–glycation activities correlates with phenolic composition of tropical medicinal herbs.Asian Pac. J. Trop. Med.,6,561-569.
39. Rao, T. P.,Sakaguchi, N.,Juneja, L. R.,Wada, E.,Yokozawa, T.(2005).Amla (Emblica officinalis Gaertn.) extracts reduce oxidative stress in streptozotocin-induced diabetic rats.J. Med. Food,8,362-368.
40. Sachan, N.,Arif, D. M.,Zaman, M. K.,Kumar, Y.(2011).Anti-inflammatory, analgesic and antioxidant potential of the stem bark of Spondias mangifera Willd.Arch. Biol. Sci.,63,413-419.
41. Sales, P. M.,Souza, P. M.,Simeoni, L. A.,Magalhães, P. O.,Silveira, D.(2012).α-Amylase inhibitors: A review of raw material and isolated compounds from plant source.J. Pharm. Pharm.,15,141-183.
42. Shimada, K.,Fujikawa, K.,Yahara, K.,Nakamura, T.(1992).Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion.J. Agric. Food Chem.,40,945-948.
43. Siddhuraju, P.,Manian, S.(2007).The antioxidant activity and free radical-scavenging capacity of dietary phenolic extracts from horse gram (Macrotyloma uniflorum(Lam.) Verdc.) seeds.Food Chem.,105,950-958.
44. Surya Prakash, D. V.,Meena, V.(2014).Optimization of physico–chemical parameters for the extraction of chebulinic acid from medicinal herbs.Banats. J. Biotechnol.,10,46-51.
45. Tsai, P. J.,Tsai, T. H.,Yu, C. H.,Ho, S. C.(2007).Comparison of NO-scavenging and NO-suppressing activities of different herbal teas with those of green tea.Food Chem.,103,181-187.
46. Wu, S. X.,Zhou, L. X.(1996).An edible value research of Phyllenthus emblica L.Academic J. Kunming Med. Coll.,17,22-23.
47. Xiang, Y.,Pei, Y.,Qu, C.,Lai, Z.,Ren, Z.,Yang, K.,Xiong, S.,Zhang, Y.,Yang, C.,Wang, D.,Liu, Q.,Kitazato, K.,Wang, Y.(2011).In vitro anti-herpes simplex virus activity of 1, 2, 4, 6-tetra-O-galloyl--β-glucose from Phyllanthus emblica L. (Euphorbiaceae).Phytother. Res.,25,975-982.
48. Yang, B.,Kortesniemi, M.,Liu, P.,Karonen, M.,Salminen, J. P.(2012).Analysis of hydrolyzable tannins and other phenolic compounds in emblic leafflower (Phyllanthus emblica L.) fruits by high performance liquid chromatography-electrospray ionization mass spectrometry.J. Agric. Food Chem.,60,8672-8683.
49. Yang, B.,Liu, P.(2014).Composition and biological activities of hydrolyzable tannins of fruits of Phyllanthus emblica.J. Agric. Food Chem.,62,529-541.
50. Yang, Y.,Lian, G.,Yu, B.(2015).Naturally occurring polyphenolic glucosidase inhibitors.Isr. J. Chem.,55,268-284.
51. Zaharudin, N.,Salmean, A. A.,Dragsted, L. O.(2018).Inhibitory effects of edible seaweeds, polyphenolics and alginates on the activities of porcine pancreatic alpha-amylase.Food Chem.,245,1196-1203.
52. Zaharudin, N.,Salmeán, A. A.,Dragsted, L. O.(2018).Inhibitory effects of edible seaweeds, polyphenolics and alginates on the activities of porcine pancreatic α-amylase.Food Chem.,245,1196-1203.