Tiiu ban Sinh hoc vd nguon lgi sinh vgt Bien
HOA^T T I N H C H O N G O X Y H O A C U A D I C H C H I E T M E T H A N O L V A C A C P H A N D O A N C H I E T D U N G M O I K H A C N H A U C U A
C H U N G TU" M O T S O L O A I R O N G B I E N V I E T N A M 'Vo Mai Nhu-Hilu. ' T r i n Nguyin Ha Vy, 'Nguyen Bach Khoa, 'Tran Thi Thanh Van
va ^Pham Quoc Long
'Vipn nghien ciiu va iing dvmg cong nghe nha hang, VKHCNVN.
2A Himg Vuong, Nha Trang, Khanh Hoa.
V i e n hoa hgc cac hgp chit thien nhien, VKHVN.
18 Hoang QuSc Vipt, C i u Giiy, Ha NOi.
Email: [email protected].
Tom tat:
Hogt tinh chong oxy hda cua dich chiet methanol vd 4 phdn dogn chiet khdc nhau:
hexan (HX). ethylaxetate (EA). butanol (Bu) vd cloroform (CF) cua 6 lodi rong bien dd dugc nghien ciru. Kit qud cho thdy cdc phdn dogn chiet dung moi khdc nhau tir dich chiet methanol cd hogt tinh chong oxy hda mgnh han dich chiet tdng. Trong so cdc phdn dogn thu dugrc thi phdn dogn Bu tir lodi rong S.crasifolium cd hogt tinh bdt gdc DPPH cao han vd phdn dogn EA tir lodi Padina australis cd hogt tinh oxi hda tdng so cao nhdt. Ket qud nghien ciru ndy dugc sir dung de nghien ciru xdc dinh, tdch chiet vd dgng cdc hgp chdt cd tinh chong oxy hda trong rong bidn.
ANTIOXIDANT PROPERTIES OF METHANOL AND ITS SOLVENT FRACTION OBTAINED FROM SELECTED VIET N A M SEAWEEDS
Abstract:
The antioxidant activities of crude methanol extract and four different solvent fractions (hexan (HX), ethyl acetate (EA), Cloroform (CF), butanol (BuOH) fro^ six selected Vietnam seaweeds were evaluated. The results indicate that different solvent fractions obtained from total (methanol) extract exhibit higher antioxidant activities as compared to the total extract. Bu fraction of S.crasifolium exhibited higher DPPH radical scavenging activity. Sc among all the fractions and EA fraction of Padina australis found to have the highest total antioxidant activity. The findings of this work are useful to further research to identify, isolate and
characterize the specific compound which is responsible for higher antioxidant activity. .
I . M d DAU
Rong bien la ngu6n cac hgp chit sinh hgc da dang va phong phu nhu carotenoid, cac axit beo, vitamin, muoi khoang ....[1]. Trong s6 cac hgp chit sinh hgc tren thi nhom hgp
Hgi nghi Khoa hgc vd Cdng ngh? bien todn qudc lan thir V
chdt chdng oxy hda cd le la nhdm hgp chat thu hiit cac nha nghien ciiu va nha sdn xudt th\rc phdm chiic nang nhit. Vi cac hgp chat chdng oxy hda dugc coi nhu la chia khoa dl chdng lai cac benh khac nhau nhu ung thu, vi6m nhiem kinh nien, xa viia dgng mach, cac benh tim mach vd ddc bigt la chdng lao hda [2]. Cac hgp chdt chdng oxy hda tir rong biin da dugc n^ien ciiu trong thdi gian gdn ddy Id phylopheophylin tu loai Eisenia bicyclls vd Porphyra sp, phlorotannins tir loai Sargassum kfellamanlanum, fiicoxanthin tir loai Hljikia fusiformis.... [3,4]
Trong bdo cao trudc chiing tdi da danh gia boat tinh chdng oxy hda cua dich chilt mdt s6 lodi rong biin sinh trudng t^i mien Trung Vigt Nam, trong bao cao ndy chung tdi tiip t\ic nghien cuu tinh chdt chdng oxy hda cua. djch chiet methanol vd 4 phan d o ^ chiet khac nhau: hexan (HX), ethylaxetate (EA), butanol (Bu) vd cloroform (CF) cua 06 loai rong biin bang cdc phuang phap thii khd nang bdt gdc DPPH, khd nang khii, boat tinh oxy hda tdng sd vd hdm lugng polyphenol trong cac phan doan.
n . NGUYEN LIEU VA PHU'ONG PHAP NGHIEN CUtJ ILl.Thu thap man rong:
Cac mdu rong bien thu thsip vao cac thang 4, 5 va 6 nam 2010 tai cac viing biin miln Trung. Rong dugc nia sach san cat va cdc chat ban ca hgc va phai khd trong bdng ram, den xay nhd va bao quan tai nhiet do phdng.
II.2.Chuan hi chiet phan doan:
lOg rong khd dugc them 200 mL methanol 50%. Qua trinh ngam ndy tiln hanh trong 2 ngay d nhiet do phdng. Sau dd, dich lgc dem cd quay d nhiet do 40^0 ttiu dugc phdn dich nude dugc chiet phan doan Idn lugt vdi cac dung mdi khdc nhau nhu hexan (HX), cloroform (CF), ethylaxetate (EA) va butanol (Bu).
n.3.Phuffng phap nghien cihi
- Xac dinh ham lugng phlorotannin theo phuang phdp so mau vdi Folin-Ciocalteu sii dyng phloroglucinol la chdt chudn [5].
- Xdc dinh boat tinh oxy hda tdng sd theo phuang phap theo phuang phdp Prieto, P va CS (1999) [6]
- Xac djnh khd nang quyet gdc DPPH theo phuang phap Yen GC va Chen HY (1995) [7]
Khd nang quet gdc DPPH (Sc) tinh theo cdng thiic :
ScVo = [1 - i^^JuZAniLi X100
A
Trong dd Ac : Do hdp thu cua mdu so sdnh (Dung dich DPPH khdng mdu); Am : Do hdp thu ciia mdu (Dung dich DPPH them vdo mlu thu): A^, : Dd hap thu ciia mdu khdng cdDPPH.
- Xac dinh kha ndng khir sdt theo phuang phap Zhu va CS (2002) [8]
Tieu ban Sinh hoc vd ngudn l^i sinh vdt Bien
III. ld:T QUA VA THAO LUAN III.L Hieu suat chiet phan doan
Bdng 1. Hiiu sudt chiit methanol (theo khdi lugng rong khd) vd hieu sudt chiet phdn dogn (theo khdi lugng chdt chiit)
STT 01 02 03 04 05 06
Ten loiii rong S.crasifolium Dictvota linearis Padina australis Turbinaria ornata Caulerpa.cup Chnoocpora implexa
Hi|u suSt Chiet ME
(%)
12,1 11,06
6,3 9,27' 11,8 5,01
Hieu suSt cac ohdn doan hexan
(HX)(%) 7,2 38,4 30,9 13,2 6,94 6,66
cloroform (CF)(%) 44,3 8,22 14,2 45,5 12,27 11,35
ethylaxetate (EA) (%)
8,6 0,7 17,0 15,15 2,11 9,87
butanol (Bu) (%)
40,8 52,7 37,8 26,06 78,6 72,0 Hieu sudt chiet methanol ciia 06 loai rong (bang 1) hong khoang tir 5,01 den 12,1 % va hieu sudt chiet cdc phdn doan cd khac biet nhau rd ret giiia cdc hg rong khdc nhau. De dang nhan thdy hieu sudt chiet phdn doan Bu cao han so vdi hau het cac phdn doan cdn l^i. Hieu suat chiet phdn doan cua 02 loai rong S.crasifolium vd Turbinaria ornata cimg hg Sargassaceae cimg cd thir tu hieu sudt cac phan doan nhu nhau vd cao nhat la phdn doan , CF tuang iing vdi gid trj 44,3 va 45,5 %. Ket qud nay tuang tii vdi nghien cim cua [9]. Hai
lodi rong Dictyota linearis va Padina australis ciing hg Dictyotaceae cimg cd hieu sudt phan do^n HX cao han cdc lodi rong khac.
III.2. Ham lugng polyphenol trong djch chiet va cac phan doan
Bdng 2. Hdm hrang polyphenol trong dich chiit methnol vd cdc phdn dogn
STT 01 02 03 04 05 06
Ten loai rong S.crasifolium Dictyota linearis Padina australis Turbinaria ornata Caulerpa.cup Chnoocpora implexa
Polyphenol trong ME mg/g
rong kho 22,23
2,0 3,4 9,35 38,6 2,6
Ham lu-Q'ng polyphenol trong cac ph hexan
(HX)(mK) 0,087 0,723 0,21 0,08 0,17 0,14
cloroform (CFKms) 0,22 0,19 0,11 0,2 0,16 0,18
ethylaxetate (EA)(mg)
3,73 0,14 0,13 2,8 0,21 0,11
an doan butanol (Bu) (mg) . 0,32
0,80 1,1 0,4 12,3
1,2 Ham lugng polyphenol trong dich chi6t methanol khac nhau theo loai, cao nhat la loai rong xanh vai ham lugng 38,6 mg/g rong kho va thap nhit la loai rong Dictyota linearis vol ham lugng 2,0 mg/g rong kho. Tuong tu nhu hieu suit chiet phan doan, ham lugng polyphenol trong cac phan doan khac nhau va cac loai rong trong cung mpt hg co cimg thii HI phan b5 ham lugng polyphenol nhu nhau. Hg rong Sargassaceae cimg co thii tu sau EA>Bu>CF>HX va hg rong Dictyotaceae co thii tu Bu>HX>CF>EA. Trong khi hai loai rong do va rong xanh ciing co ham lugng polphenol trong phan doan Bu la cao nhat.
Hdi nghj Khoa hgcvd Cdng nghe bien todn qudc Idn thu V
ni.3. Hoat tinh oxy hda tdng so cua djch chiet va cac phan doan
Bdng 3. Hogt tinh chdng o.xy hoa cua dich chiit methanol vd cdc phdn dogn tinh theo mg AscB/g chdt chiet khd
STT 01 02 03 04 05 06
Ten loai rong S.crasifolium Dictyota linearis Padina australis Turbinaria ornata CaUlerpa.cup Chnoocpora implexa
Hoat tinh chong oxi hoa tong dich chilt ME(mg)
110 166,9 72,1 65,1 94,06 30,7
Hoat tinh chong oxi hoa cac phan doan mg ascb hexan
(HX)(mg) 94,0 206,7 123,6 58.1 170,0 154,5
cloroform (CF)(mg) 112,2 248,4 1340 149,3 451,4 853,0
ethylaxetate (EA)(mg)
206,3 151,6 1700 408,0 955,8 101,7
butanol (Bu) (mg)
213,0 190,1 115,0 300 141,9 110,1 Tii sd lieu bang 3 cho thay rang boat tinh chdng oxy hda tdng sd cua cdc phdn dosm chilt khdc nhau cao ban boat tinh cua phdn doan chiet thd mac du phdn doan chiet tho chiia tat ca cdc hgp chdt chdng oxy hda khdc nhau. Yuan va CS [10] cho rang mgt sd dung mdi cd dnh hudng den dang ciia cdc hgp chat hda hgc va chinh mgt sd hgp chdt hda hgc nay dd can trd boat tinh chdng oxy hda'trong dich chiet methanol. Cac loai rong S.crasifolium va Turbinaria ornata cimg cd thii tu hoat tinh oxy hda tang din ciia cac phan doan nhu sau: EA>Bu>CF>ME>HX. Cdc phdn doan CF ciia rong Chnoocpora implexa va phdn doan EA cua rong Caulerpa.cup va dac biet la ed 02 ^hdn doan CF va EA cua loai rong Padina australis cd hoat tinh chdng oxy hda cao gdn bang hoac ban vitamin C, trong khi theo sd lieu bang 2 thi hdm lugng polyphenol khdng cao so vdi cac phan do?n khdc.
Ket qua nay chiing td rang ngoai polyphenol cdn cd mat cac cdc chdt chdng oxy hda khac trong nhiing phdn doan nay cd die la nhdm cac chat chuyen hda thii cap nhu sesquiterpenoid, diterpenoid [11]. Phdn doan Bu cua rong Caulerpa.cup cd hdm lugng polyphenol cao nhat (Bang 2) nhimg hoat tinh chdng oxy hda khdng cao. Dieu nay dugc giai thich nhu sau cac chdt thugc nhdm hgp chdt polyphenol cd nhieu dang cdu tnic khac' nhau vd boat tinb chdng oxy hda phu thudc vdo cau tnic cua chiing, vi vdy md trong phdn doan Bu cd the cd nhieu hgp chdt polyphenol cd dang cdu tnic gdn nhau vd d?ng cdu tnic nay cd hoat tinh chdng oxy hda khdng cao.
in.4.Hoat tinh khu sat (Fe) cua djch chlet va cac phan doan
Bdng 4. Hogt tinh khu sdt (Fe) cua dich chiet methanol vd cdc phdn dogn tinh theomg AscB/g chdt chiet kho
STT 1 2 3 4 5 6
Ten loai rong S.crasifolium Dictvota linearis Padina australis Turbinaria ornata Caulerpa.cup Chnoocpora implexa
Kha nang khir sat chiet
ciia ME 72 86,3 80,5 79,8 80,5 56,0
Kha nang khir sat hexan
(HX) 88,9 77,8 98,2 59,3 91,0 197,7
cloroform (CF) 104,4 54,7 900,5
65,3 240.4 366,7
cac phSn doan mg Fe ethylaxetate
(EA) 50,8 126,6 1966 75,2 285,7 107,7
butanol (Bu) 343,2 71,5 71,9 126,2 709,1 •
72,3
Tieu ban Sinh hgc vd nmdn lgi sinh vdt Biin
^ Tu so lieu bang 3 va 4, de dang nhan thiy ring cac phan doan cd hoat tinh ch6ng oxy hoa cao cung la cac phSn doan co ho?t tmh khii sit cao nhu la cac phan doan CF va EA cua rong Padina australis, phan dojn CF cua rong Chnoocpora implexa . Khac vai hojt tinh chong oxy hoa phan doan Bu cila rong Caulerpa.cup vira c6 ham lugng polyphenol cao vira CO hoat tinh khu sit cao. Ket qua nay co thS do cac hgp chit polyphenol dugc chiet bang dung moi co cau tnjc c6 hoat tinh khii cao.
in.S.Kha nang bat goc DPPH ciia dich chiet va cac phSn doan
Goc tii do (ROS) thuang kem bSn va co boat tinh cao, phSn hi cua chiing thuong chiia mpt electron ty: do ho}c khong can bing dien hi, chiing dugc tao ra trong co thi d8 ch5ng lai cac vi khuari, vi nit gay benh tuy nhien khi cr ham lugng cao cac g6c tu do nay phan img vai protein, lipid, ADN va gay ra mgt s6 benh nghiem trgng nhu: ung thu, suy than, benh tim mach. Vi vay ma mgt honj cac thong so quan hgng dJ xac dinh hoat tinh ch6ng oxy hoa la xac djnh kha nang quet goc DPPH.
Ket qua so lieu trong bang 5 cho thiy trong s6 06 dich chiSt methanol cua 06 loai rong thi kha nang bat g6c DPPH ciia loai rong xanh Chnoocpora implexa la cao nhit va thap nhat la 02 loai rong nSu Dictyota linearis va Padina australis ciia cimg hg Dictyotaceae vai cac gia hi EC50 tucmg iing la 0,26 va 14,2 ; 12,12 mg chit kho. Dai vdi kha nang bit goc DPPH ciia djch chiet cr cac phan doan khac nhau thi phSn doan Bu cua rong S.crasifolium la cao nhit vai EC50 la 0,085 mg chit kho va phSn doan HX, CF cua rong Dictyota linearis la thap nhat vol gia tri EC50 la 20,5 mg chit kho.
Bdng 5. Khd ndng bdt goc DPPH ciia dich chiet methanol vd cdc phdn dogn tinh theg ECso (mg chdt chiet khd)
STT 1 2 3 4 5 6
Ten loai rong S.crasifolium Dictyota linearis Padina australis Turbinaria ornata Caulerpa.cup Chnoocpora implexa
Kha nang bat goc DPPH djch
chiet ME 1,08 14,2 12,12 0,94 1,91 0,26
Kha nang b i t goc DPPH cac phan doan EC50 (mg) hexan
(HX) 0,11 20,5 0,44 1,2 0,77 6,95
cloroform (CF)
0,9 20,5 1,32 6,15 2,5 2,5
ethylaxetate (EA)
1,16 2,2 1,0 2,45 1,18 2,66
butanol (Bu) 0,085 16,59 13,35 0,48 1,0 0,76 Dieu thii vi la kha nang bat gdc DPPH ciia cimg phan d o ^ EA tir cac lodi rong khdc nhau thay ddi khdng ddng ke trong khi ciing cdc phdn doan HX, CF va Bu td cac loai rong khac nhau cd su khac nhau nhieu. Cimg hg rong Dictyotacea, cung cd dich chilt ME, cung cd khd nang bat gdc DPPH thap nhung kha ndng bat gdc dxt phdn doan HX va CF cua rong Padina australis cao ban nhieu so vdi rong Dictyota linearis.
Khd ndng bdt gdc DPPH khac nhau d cac lodi rong khac nhau cd the do thdnh phdn hgp chdt cd boat tinh sinh hgc khac nhau. Loai rong S.crasifolium, Caulerpa.cup vd Chnoocpora implexa cd the chiet dugc nhieu chdt cd boat tinh smh hgc d phan doan dung mdi phdn c\rc Butanol. Ngugc lai, dung mdi khdng phdn cue hexan va it phdn cue clorform cd thi chiet dugc nhieu chdt cd khd nang bit gdc DPPH d loai rong S.crasifolium, Padina australis va Caulerpa.cup.
Hgi nghi Khoa hgc vd Cdng nghg bien todn qudc lan thuY
IV. KET LUAN
Dich chiSt Metanol va cac phan doan HX, CF, EA va Bu ciia 6 loai rong ( 4 loai rong nau, 1 loai rong do va 1 loai rong xanh) deu the hien hoat tinh chong oxi hoa. Rong S.crasifolium the hien kha nang co hoat tinh dang kS nhit. Tiiy thugc vao loai rong the hien hoat tinh ch6ng oxi hoa khac nhau a nhirng phan doan khac nhau. Ket qua nghien ciiu nay dugc su dimg dl nghien ciiu xac dinh, tach chiet va dang cac hgp chat co tinh chong oxy hoa trong rong bien.
LOfI CAM CJN
Tap thi tac gia xin cam an chuang trinh nghien ciiu ca ban djnh huang iing dung - Bp KH & CN (Hgp d5ng s6 : 05/20I0/HD-NCCB UD) da h5 hg kinh phi di thuc hien bai bao.
TAI LIEU THAM KHAO 1. Fleurence, J.,. Trends Food Sci. Technol. 10,25-28 (1999).
2. Kohen, R., & Nyska, A. (2002). Toxicology Pathology, 30(6), 620-650 (2002).
3. Sachindra, N.M, Sato E., Maeda H., Hosokawa M., Niwano, Y., Kohno M & Miyashita K. J. Agric Food chem 55: 85-16-8522 (2007).
4. Yabuta, Y, Fujisnura, H., Kwa C.S., Enomoto & Watrnabe, F.. Food Sci Technol. Res.
16:347_351 (2010).
5. Velioglu YS, Mazza G, Gao L, Oomah BD, J Agric Food Chem 46:4113^117 (1998) 6. Prieto, P., Pineda, M., Aguilar, M., Anal. Biochem. 269,337-341 (1999) 7. Yen GC, Chen HY, J Agric Food Chem 43:27-37 (1995).
8. Zhu, Q.Y., Hackman, R.M., Ensunsa, J.L., Holt, R.R., Keen, C.L., J Agric Food Chem 50, 6929-6934. (2002)
9. Hong Ye, Chunhong Zhou, Yi Sun, Xin Zhang, Jim Liu, Eur Food Res Technol 230:101-109(2009).
10. Yuan, Y.V., Bone, D.E., Carrington, M.F.,. Food Chem. 91,485^94 (2005).
11. Fenioal W, Paul VJ,. Hydrobiologia 116/117:135-140 (1984)
