Phn lp cc hp cht phenolic t mt s thc vt Vit Nam

Nguyn Th Qunh Hoa

Trng i hc Khoa hc T nhin Khoa Ha hc, Chuyn ngnh: Ha hu c, M s: 60 44 27 Ngi hng dn: PGS. TS. Phan Minh Giang Nm bo v: 2012
Abstract: Trnh by tng quan: cc hp cht phenolic thc vt, gii thiu v cy Cho l phong (Engelhardtia spicata Lesh ex. Blume). Gii thiu nhim v v phng php nghin cu: phng php phn tch, phn tch cc hn hp v phn lp cc hp cht, phng php xc nh cu trc. Trnh by kt qu: quy trnh chit cc phn chit t cy Cho l phong, phn tch phn chit etyl axetat (EG3), phn tch phn chit nc (EG4), cu trc cc hp cht c phn lp. Tin hnh thc nghim: thit b v ha cht, nguyn liu thc vt, iu ch cc phn chit t cy Cho l phong, phn tch v phn tch phn chit etyl axetat (EG3), phn tch sc k ct phn chit nc (EG4), hng s vt l v d kin ph ca cc hp cht c phn lp. Keywords: Ha hu c; Qa trnh phn lp; Hp cht phenolic; Thc vt Content Cc hp cht phenolic thc vt nh cc axit phenolic, cc flavonoit v cc flavomoit polyme ang ngy cng thu ht c nhiu s quan tm do tnh cht chng oxi ha, tc dng phng nga ung th v cc bnh lin quan n tim mch ca cc hp cht phenolic. Cc nghin cu (ha hc, dc l hc, lm sng) phn no l gii c mi lin quan gia sc khe con ngi v vic tiu th cc sn phm thc phm giu cc hot cht phenolic thin nhin. Cc hp cht phenolic chim mt v tr ng k trong s cc nhm hp cht thin nhin c tc dng d phng ung th. Chng c cu trc a dng v xut hin ph bin trong gii thc vt. Vic phn lp cc hp cht ny cho cc th nghim hot tnh sinh hc c th c thc hin bng cc phng php chit v sc k iu ch; tuy nhin ph rng tan ca cc hp cht ny cho thy mi qui trnh phn lp nn c gii hn vo mt nhm hp cht phenolic. Cc sng lc sc k lp mng s b xc nh c nhm hp cht phenolic trong cc phn chit t cc loi cy: Alnus nepalensis D. Don (Betulaceae), Betula alnoides Buch. Ham. ex D. Don (Betulaceae) v Engelhardtia spicata Lesch. ex. Blume (Juglandaceae). Mc tiu nghin cu ca lun vn ny l xy dng mt qui trnh chit cc phn chit giu cc hp cht phenolic v phn lp sc k cc hp cht phenolic, sau cu trc chnh xc ca cc cc hp cht c phn lp s c xc nh bng cc phng php ph hin i. Qui trnh ny c p dng thnh cng phn lp cc hp cht phenolic thnh phn chnh t l cy Cho l phong (E. spicata).

Chng 1 TNG QUAN 1.1 Cc hp cht phenolic thc vt 1

1.1.1 Gii thiu v cc hp cht phenolic thc vt [6] Cc hp cht phenolic l cc hp cht c mt hoc nhiu vng thm vi mt hoc nhiu nhm hydroxy. Chng c phn b rng ri trong gii thc vt v l cc sn phm trao i cht phong ph ca thc vt. Hn 8.000 cu trc phenolic c tm thy, t cc phn t n gin nh cc axit phenolic n cc cht polyme nh tannin. Cc hp cht phenolic thc vt c tc dng chng li bc x tia cc tm hoc ngn chn cc tc nhn gy bnh, k sinh trng v ng vt n tht, cng nh lm tng cc mu sc ca thc vt. Chng c khp cc b phn ca cy v v vy, chng cng l mt phn khng th thiu trong ch n ung ca con ngi. 1.1.2 Phn loi cc hp cht phenolic [6, 10] Cc hp cht phenolic c cu trc rt a dng v c th c chia thnh 10 nhm chnh. Cc hp cht phenolic thc vt bao gm cc stilben, cc lignan, cc axit phenolic, cc flavonoid v cc tannin. 1.1.3 Hot tnh sinh hc ca cc hp cht phenolic Do s phn b rng ri, cc polyphenol c vai tr i vi sc khe ca con ngi nn ch n ung dinh dng c ch trong nhng nm gn y. Cc nh nghin cu v cc nh sn xut thc phm tp trung vo cc polyphenol c c tnh chng oxy ha mnh trong ch n ung, cc hiu ng ng tin cy ca chng trong vic phng nga nhng chng bnh cng thng oxy ha lin quan.Theo cc nghin cu dch t hc, hp th cc hp cht phenolic s gim c nguy c mc cc bnh tim mch ngn nga c bnh ung th. Hn na cc polyphenol cn c cc tc dng sin h l hc c th trong vic ngn nga v iu tr bnh. 1.1.4 Phng php chit v phn lp cc hp cht phenolic 1.1.4.1 Phng php chit 1.1.4.2. Phng php phn lp v tinh ch 1.1.4.3 Phn tch v xc nh cu trc 1.2 Gii thiu v cy Cho l phong (Engelhardtia spicata Lesh ex. Blume) 1.2.1 c im thc vt hc 1.2.2 Ni sng v thu hi 1.2.3 Cng dng ca cy Cho l phong 1.2.4 Mt s nghin cu v thnh phn ha hc v hot tnh sinh hc ca chi Engelhardia (Juglandaceae)

Chng 2 NHIM V V PHNG PHP NGHIN CU 2.1 Nhim v ca Lun vn 1) Xy dng quy trnh chit v phn lp cc hp cht phenolic phn cc t l cy Cho l phong; 2) Kho st sc k lp mng cc phn chit cha cc hp cht phenolic nhn c. nh gi nh tnh v s phn gii, kh nng phn tch cc phn chit v phn lp cc hp cht thnh phn; 3) Xy dng quy trnh phn tch cc phn chit phn lp cc hp cht phenolic thnh phn chnh trong l cy Cho l phong; 4) Xc nh cu trc cc hp cht c phn lp. 2.2 Phng php nhin cu 2.2.1 Cc phng php phn tch, phn tch cc hn hp v phn lp cc hp cht 2.2.2 Cc ph-ng php xc nh cu trc Chng 3 KT QU V THO LUN 3.1 i tng nghin cu L cy Cho l phong (Engelhardtia spicata Lesch. ex Blume, Juglandaceae) c thu hi ti Tam Sn, Ph Th vo thng 2 nm 2008. 3.2 Quy trnh chit cc phn chit t l cy Cho l phong 3.3 Phn tch phn chit etyl axetat (EG3) 3.4 Phn tch phn chit nc (EG4) 3.5 Cu trc cc hp cht c phn lp 3,5,7-Trihydroxychromon (cht I) Aromadendrin (II) 3,5,7-Trihydroxychromon 3-O- -L-rhamopyanosid (Eucryphin) (cht III) Taxifolin 3-O- -L-rhamnopyranosid (Astilbin) (IV Chng 4 THC NGHIM 4.1 Thit b v ha cht 4.2 Nguyn liu thc vt L cy Cho l phong (Engelhardtia spicata Lesch. ex Blume, Juglandaceae) c thu thp vo thng 11 nm 2007 ti Tam Sn, Ph Th. 4.3 iu ch cc phn chit t cy Cho l phong 4.4 Phn tch v phn tch phn chit etyl axetat (EG3) 4.4.1 Phn tch sc k lp mng phn chit etyl axetat (EG3) 4.5 Phn tch sc k ct phn chit nc (EG4) 4.5.1 Phn tch sc k lp mng phn chit nc (EG4) 4.5.2 Phn tch phn chit nc (EG4) 4.6 Hng s vt l v d kin ph ca cc hp cht c phn lp KT LUN 4

Lun vn tt nghip Nghin cu phn lp cc hp cht phenolic t mt s loi thc vt ca Vit Nam thc hin cc nhim v nghin cu phn lp cc hp cht phenolic t l cy Cho l phong. Lun vn hon thnh cc nhim v nghin cu v t c cc kt qu chnh sau: 1. xy dng c qui trnh chit cc hp cht phenolic vo phn chit etyl axetat (EG3), hiu sut chit 0,5% so vi lng nguyn liu kh) v phn chit nc ( EG 4) t l cy Cho l phong. 2. phn tch sc k lp mng (TLC) cc phn chit etyl axetat v phn chit nc xc nh iu kin sc k phn tch nh tnh, xc nh cc h dung mi thch hp cho phn tch sc k iu ch gradient cc phn chit phenolic v kim tra cc qu trnh phn tch. 3. Bng cc qui trnh phn tch cc hp cht phenolic phn cc kt hp sc k ct thng (CC) gradient, sc k ct tinh ch (Mini-C) v kt tinh li phn lp c 2 hp cht chromon v 2 hp cht flavonoit t cc phn chit etyl axetat v phn chit nc. 4. S dng cc phng php ph hin i ESI-MS, 1H-NMR, 13C-NMR v DEPT xc nh c cu trc ca cc hp cht phenolic thnh phn chnh ca l cy Cho l phong l 3,5,7-trihydroxychromon (I), aromadendrin (II), 3,5,7-trihydroxychromon 3O- -L-rhamopyanosid (eucryphin) (III) v taxifolin 3-O- -L-rhamnopyranosid (astilbin) (IV). Astilbin c chng t l mt hp cht c tc dng chng oxi ha, lm gim nng cholesterol v lipid trong gan.

References Ting Vit 1. V Vn Chi (1997), T in cy thuc Vit Nam, Nh xut bn Y hc, Thnh ph H Ch Minh, tr. 226. Ting Anh 2. Agarwal, C., Veluri, R., Kaur, M., Chou, S. C., Thompson, J. A.; Agarwal, R. (2007),Fractionation of high molecular weight tannins in grape seed extract and identification of procyanidin B2-3,3'-di-O-gallate as a major active constituent causing growth inhibition and apoptotic death of DU145 human prostate carcinoma cells, Carcinogenesis, 28, pp. 1478-1484. 3. Asquith, T. N., Izuno, C. C., Butler, L. G. (1983), Characterization of the condensed tannin (proanthocyanidin) from a group II sorghum, J. Agric. Food Chem, 31, pp. 1299-1303. 4. Castro-Vargas, H.I., Rodrguez-Varela, L.I., Ferreira, S.R.S., Parada-Alfonso, F. (2010), Extraction of phenolic fraction from guava seeds (Psidium guajava L.) using supercritical carbon dioxide and co-solvents, J. Supercrit. Fluid., 51, pp. 319-324. 5. Chu, T. Y., Chang, C. H., Liao, Y. C., Chen, Y. C. (2001), Microwave-accelerated derivatization processes for the determination of phenolic acids by gas chromatography-mass spectrometry, Talanta, 54, pp. 1163-1171. 6. Dai, J.; Mumper, R. J. (2010), Plant Phenolics: Extraction, Analysis and Their Antioxidant and Anticancer Properties, Molecules, 15, pp. 7313-7352. 2

7. De Paule M. Moreira, F., Coutinho, V., Montanher, A. B. P., Caro, M. S. B., Brighente,
I. M. C.; Pizzolatti, M. G. (2003), Flavonoids and Triterpenes from Bacchsis psendotenui folia - Bioactivity on Arternisia salina, Quim. Nova, 26, pp. 309-311. 8. Dobi, P., Pavlkov, P., Adam, M., Eisner, A., Beov, B., Ventura, K. (2010), Comparison of pressurised fluid and ultrasonic extraction methods for analysis of plant antioxidants and their antioxidant capacity, Cent. Eur. J. Chem, 8, pp. 87-95. 9. Ek, S., Kartimo, H., Mattila, S., Tolonen, A. (2006), Characterization of phenolic compounds from lingonberry (Vaccinium vitis-idaea), J. Agric. Food Chem, 54, pp. 9834-9842. 10. Garcia-Salas, P., Morales-Soto, A., Segura-Carretero, A., Fernndez-Gutirrez, A. (2010), Phenolic-Compound-Extraction Systems for Fruit and Vegetable Samples, Molecules, 15, pp. 8813-8826. 11. Gmez, A. M, Carrasco, A, Caabate, B, Segura, A, Fernndez, A. (2005), Electrophoretic identification and quantitation of compounds in the polyphenolic fraction of extra-virgin olive oil, Electrophoresis, 26, 3538-3551. 12. Hagerman, A. E., Butler, L. G. (1980), Condensed tannin purification and characterization of tannin-associated proteins J. Agric. Food Chem, 28, pp. 947-952. 13. Igarashi, K., Uchida, Y., Murakami, N., Mizutani, K., Masuda, H. (1996), Effect of astilbin in tea processed from leaves of Engelhardtia chrysolepis on the serum and liver lipid concentrations and on the erythrocyte and liver antioxidative enzyme activities of rats, Bioscience Biotechnology and Biochemistry, 60, pp. 513-515. 14. Jac, P., Polasek, M., Pospisilova, M. (2006), Recent trends in the determination of polyphenols by electromigration methods. J. Pharm. Biomed. Anal, 40, pp. 805-814. 15. Juregui, O., Galceran, M. T. (2001), Handbook of Analytical Separations; University of Barcelona, Barcelona, Spain, Chapter 6, pp. 196. 16. Kaehkoenen, M. P., Heinaemaeki, J., Ollilainen, V., Heinonen, M. (2003), Berry anthocyanins: Isolation, identification and antioxidant activities, J. Sci. Food Agric, 83, pp. 1403-1411. 17. Kasai, R.; Hirono, S., Chou, W. H., Tanaka, O., Chen, F. H. (1988), Sweet dihydro flavonol rhamnofide from leaves of Engelhardtia chrysolepis, a Chinese folk medicine, Hung-gi, Chem. Pharm. Bull., 36, pp. 4167-4170. 18. Klejdusa, B., Kopeck, J., Benesov, L., Vaceka, J. (2009), Solidphase/supercritical-fluid extraction for liquid chromatography of phenolic compounds in freshwater microalgae and selected cyanobacterial species, J. Chromatogr. A, 1216, pp. 763-771. 19. Liazid, A., Palma, M., Brigui, J., Barroso, G. C. (2007), Investigation on phenolic compounds stability during microwave-assisted extraction, J. Chromatogr. A, 1140, pp. 29-34. 20. Llorach, R.; Gil-Izquierdo, A., Ferreres, F., Tomas-Barberan, F. A. (2003), HPLCDAD MS/MS ESI characterization of unusual highly glycosylated acylated flavonoids from cauliflower (Brassica oleracea L. var. botrytis) agroindustrial byproducts, J. Agric. Food Chem, 51, pp. 3895-3899. 21. Lin, Y., Zhang, C., Zhang, M. (2009), Chemical constituents in herbs of Polygonum jucundum, China Journal of Chinese Materia Medica, 34, pp. 1690-1691. 22. Liu, H.; Wang, S., Cai, B., Yao, X. (2004), Anticancer activity of compounds isolated from Engelhardtia serrata stem bark, Archives of Physiology and Biochemistry, 42, pp. 475-477. 3

23. Lcia Q. A. Galotta, A., Amlia D. Boaventura, M., A. R. S. Lima, L. (2008),

Antioxidant and cytotoxic activities of Euterpe precatoria Mart. Quim. Nova, 31, pp. 1427-1430. 24. Mahugo, C., Sosa, Z., Torres, M.E., Santana, J.J. (2009) Methodologies for the extraction of phenolic compounds from environmental samples: new approaches, Molecules, 14, pp. 298-320. 25. Naczk, M., Shahidi, F. (2006), Phenolics in cereals, fruits and vegetables: occurrence, extraction and analysis J. Pharm. Biomed. Anal, 41, pp. 1523-1542. 26. Palma, M., Pieiro, Z., Barroso, C.G. (2002), In-line pressurized-fluid extractionsolid-phase extraction for determining phenolic compounds in grapes, J. Chromatogr. A, 968, pp. 1-6. 27. Picouet, P. A., Landl, A., Abadias, M., Castellari, M., Vias, I. (2009), Minimal processing of a Granny Smith apple pure by microwave heating, Innov. Food Sci. Emerg, 10, pp. 545-550. 28. Pinelo, M.; Laurie, V.F., Waterhouse, A.L. (2006), A simple method to separate red wine nonpolymeric and polymeric phenols by solid-phase extraction, J. Agric. Food Chem, 54, pp. 2839-2844. 29. Ranilla, L. G., Genovese, M. I., Lajolo, F. M (2007), Polyphenols and antioxidant capacity of seed coat and cotyledon from Brazilian and Peruvian bean cultivars (Phaseolus vulgaris L.), J. Agric. Food Chem, 55, pp. 90-98. 30. Ray Sahelian, M. D. (2006), Polyphenols supplement research study, health benefit. 31. Ross, K. A.; Beta, T.; Arntfield, S. D. (2009), A comparative study on the phenolic acids identified and quantified in dry beans using HPLC as affected by different extraction and hydrolysis methods, Food Chem, 113, pp. 336- 344. 32. Vilkhu, K.; Mawson, R., Simons, L., Bates, D. (2008), Applications and opportunities for ultrasound assisted extraction in the food industry, Innov. Food Sci. Emerg., 9, pp. 161-169. 33. Wu, C. C.; Peng, C. F.; Tsai, I. L.; Abd El-Razek M. H.; Huang H. S.; Chen I. S. (2007), Secondary metabolites from the roots of Engelghardia roxburghiana and their antitubercular activities Phytochemistry, 68, pp. 1338-1343. 34. Zhang, Y., Seeram, N.P., Lee, R., Feng, L., Heber, D. (2008), Isolation and identification of strawberry phenolics with antioxidant and human cancer cell antiproliferative properties. J. Agric. Food Chem., 56, pp. 670-675.