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OAlXIH GIA H A M LU01VG POLYPHEIVOL, DIEP LIJC VOI HOAT TilXIH CHdlVG OXY HOA Q CAC PHAI\I DOAIV
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Bai bao nay trinh bay k^t qua nghifin ciiu v^ cong doan 1 ^ sach polyphenol, clorophyl (di^p luc) v6i hoat tinh chdng oxy hoa c6 nguon gdc tu cSy ng6. n-hexan, ethanol, ethyl acetate duoc su dung nhu dung moi d^
phan dogn lam sach. H^m lugmg polyphenol, clorophyl voi hogt tinh chdng oxy hoa (tdng, khu s^t vi DPPH) a timg phln cky ng6 da duoc danh giA. K^t qua cho thay, ham luong polyphenol 6- than cSy ngo la nhi^u nhat, ham lugng clorophyl voi hoat tinh chdng oxy hoa tdng vt khii sit oia c^y ng6 cao nhit. DPPH ciia la va rS cay ng6 tdt nhat Phan doan etyla axetat cho ham luong polyphenol va hoat tinh chdng oxy hoa la cao nhat.
Tii khda: Polyphenol di$piuc, chdng oxy hoa, cay ngo, phan doan.
I.RMrDAU
Cay ngo 11 mpt trong 5 loai cay luong thuc pho hien cr ViSt Nam cdng nhu tr^n th^ gioi. Tinh d^n nam 2013, tong di6n tieh trdng ngo 6 Viet Nam da 11 1,1 - 1,3 tri$u ha, vol ntog suat binh quan 4,5 tan/ ha (Trung ttol Tin hpc va Thong k& - Bo Nong nghidp vl Phit trien Nong thon, 2013). Cay ngd chiia rlt nhi^u boat chat sinh hpe nhu: polyphenol (hgnin, tannin,...), clorophyl, lyzin, vitamin,... vl trong y hpc cd truyan chiing duoc iing dung dieu tri cic benh khic nhau nhir soi than, soi bing quang, chiing ra nhi^u mo hoi, gan, than,...(ft^.'//
vaas.vn/kienthuc/cayngo/Monantungolathuocchua benh.php.). Nhi^u tli lidu chiing minh, clorophyl va polyphenol chiem khoang 11 - 26% khdi lupng khd eua timg phln ely ngo (Rebecca va cs, 2009; Samuel
vi cs, 2011). Dong thdi polyphenol, clorophyl ciing duoc chiing minh ring co nhi^u cdng dung khie nhau trong y hpc nhu: khtog khuan, ehdng oxy hoa, khtog ung thu, lpc mlu,.,. (Levent, 2011; Stefano, 2009) vl dong vai tro nhu ehlt mlu trong thuc pham
(Stefano, 2009). Nhu vly, cay ngo 11 nguon cung cap polyphenol, clorophyl gilu co eho ngtoh cong nghifip thue pham vl y dupc.
Hien nay, hfin the gioi cung rlt it cle cdng hinh nghifin eiiu ve qui hinh tieh chi^t, Itoi saeh va nghifin Clhl sau ban v6 cle ung dung eua polyphenol,
' Vien Nghien cihi va U'ng dung Cong nghe Nha Trang, VAST
^ Cue Ch^ biSn, Thuang mgi nong lam thuy san va Nghg mu6i - Bo NN&PTNT
clorophyl eo ngudn gdc tu cly ngo de iing dung trong cupc sdng. O Vifit Nam, chua eo edng tiinh nghifin ciiu nio ve polyphenol, clorophyl vm ho^t tinh ehdng oxy hoa tu cky ngd. Chu yeu tie dung sinh hpc cua cay ngo dupc trinh biy o- y hpc d truyen.
Do do, bai blo nay trinh bay k^t qui nghien c^
v^ eong doan Ilm saeh polyphenol, elorophl voi hoat tinh chdng oxy hoa a cle phto doto khic nhau, ii dinh hudng tinh saeh polyphenol, clorophyl vdi hoai tinh ehdng oxy boa iing dung trong thuc pham chiic ntog vl dupe pham.
2. NGUYBU LEU VA PHUDNG PHAP NGHIBU CUU
2.1. Ddi hrpng, thu mlu vl blo quin mlu Gidng ngo lai sd 1 trdng tai Di6n Khtoh, Khanh Hda duoc thu mlu vao thdi diem 75 ngay. Mlu duoc tach hat de lly rfi, thto, la, rto va loi ngo. Tat cl cac mlu da loai hat dupc say l$nh a 50''C den dp am 149t Sau khi sly lanh, mlu dupc bao quto trong phoi^
lanh 4''C.
2.2. Phuong p h ^ nghifen cuu
2.2.L Bd tri thi nghi&m va chuan hi miu Trudc khi chiet, mlu dupc ng^m 15 phiit hong nude rdi vat ra dg rIo 5 phiit. Sau dd mlu duoc xay nho d^n kich thuoc 2 - 4 mm. Him luong MgCOj dupe bo sung trong qui trinh xu 1^ mlu 11 0,04%. Tat ca cle mlu deu dupe chi^t a nhifit dp phong, thffl gian 24 gio vdi ty le dung moi/ nguyfin lifiu 11 20/1 (v/vif). Dich ehiet thu dupc lpe qua ming lpc. Dich
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sau Ipc duoc danh gil ham luong polyphenol, clorophyl vcri boat tinh ehdng oxy hda.
2.2.2. Phuongphip dinh luang
Dinh lupng polyphenol tdng CTPC) theo phuong phip Folin-Ciocalteu nhu md ta boi Swanson va es (2002). Phlorotamn tdng (TPC) theo phuong phip Folin-Ciocalteu nhu md t l bdi Swanson eung cdng su (2002) vdi phlorogluemol 11 ehlt ehuan. Lly 300 ^il dich mlu bd sung 01 ml Fohn-Ciocalteu 10%, giii 5 phiit. Tiep theo thfim vio 2 ml NaaCOj 10%, frdn d^u, giu 90 phiit trong bong tdi, sau do do dp h i p thu o budc sdng 750 nm, do trfin may UV-Vis Spech-ophotometer JenWay 6400/ 6405.
Dinh lupng clorophyl tdng tiieo lichtenthaler v l cs (2001).
2.2.3. Phuong phip xic dinh hoat tinh
• Chdng oxy hda tdng (Ti^ dupc xle dinh theo phuong phip eua Prieto va cs (1^9), lly lOOpl mlu bo sung 900 pi nudc cat v l thfim 3 ml dung dich A (HaSO^ 0,6 M, phdt phat nahi 28 mM vl moHpdat amon 4 mM). Hdn hpp dupc giu 90 phut a QS^C. Sau dd do b bude song 695 nm vdi chat chuan la axit ascobic.
- Khii Fe (RP) dupc xic dinh theo phuang phap cua Zhu vl es (2002), lly 500 pi dich mlu bd sung 0,5 ml dem phdt phat pH = 7,2 v l 0,2 ml K3tFe(CN)6] 1%.
Giii hdn hpp 20 phut d 50''C. Sau dd thfim vio 500 pi CCI3COOH 10% vdi su bd sung 300 pi nude cat va 80 pi FeClg 0,1%. Tiep theo do a buoc song 655 nm vol chit chuan 11 FeSO*.
- Bit gde tu do DPPH ti^n htoh theo Blois va es (1958), lly lln lupt 200 pi, 400 pi, 600 pi, 800 ul vl 1000 pi dieh chidt vio 5 dng nghi&m, rdi bd sung 3 ml DPPH (25 mg/l) vio timg dng nghiem Ilm dung dich mlu (mlu). 6 dung dich trang (mlu tring) Itoi tuong tu mlu nhung thay DPPH bang 3 ml cdn tuyfit ddi vao tiing dng. Mlu kiem soat chuan hi bing cleh Ilm gidng nhu mlu trang nhung thay dieh chiet bang DPPH. Giii cac hdn hpp trong tdi d- nhiet dd phdng.
Sau 30 phut tidn htoh do o budc sdng 550 nm. Cdng thuc tinh phln trtoi bat gdc nhu sau:
AV^-- xlOO%
2.2.4. Phin tich ddlieu
TTiuc nghifim duac tien htoh lap lai 3 lln (n=3), phto tieh ANOVA, hoi quy tren co sd su dung phan mem MS. Excel 2010.
3.KFrqUAVATHA0LUAIV
3.1. Him lupng p o ^ h e n o l , clorophyl vdi ho?it tinh ehdng oxy hda d c l cly ngd v l tiing bd ph§n trfin cly ngfi
Sau khi chiet mlu b i n g ethanol 96%, dich chidt thu dupe tifin htoh cd dac v l phto doan d^ thu diroc him lupng clorophyl v l polyphenol cd boat tinh chdng oxy hda cao. Ket qui thu dupe nhu sau:
Ham lupng polyphenol hda tan 6 thto c^y ngd cao hon so vdi elc bO phan khic cua cly ngd, iing vdi 2,41 ± 0,01 g axit gaUc/100 g mlu kho. Hoat tinh ehdng oxy hda tdng v l khii sit d thto tuong iing 5,354 ± 0,02 g axit ascobic/ lOOg mlu kho v l 11,762
± 0,01 g FeSOyiOO g mlu kho (Hinh 1 va 2).
_ ^ ^ •HJmkftDg polyphenol (^lOOg mau kho]
Q Him tucmg chkm^hyll (mg/g mau kbo)
> 2.00
a 1 ^ S. 1,00 l 0 , 5 0 a 3 0,00
Hinh 1: Him lupng polyphenol, clorophyl trfin tong bo p h | n ciJia cly ngd
H Hoai tnh cbSng oxy hoa t6ng{g acid ascotc/IOOg mau tiro) B Ho?t tiih khir sit (g FeS04/IOOg nJu kho)
11,762 ±0,IW2
9.S70± 0,002 7^62 ±0,003
73 ''366*
May UV-Vis Spectrophotometer JenWay 6400/
3405 dupe su dung d^ do elc mlu.
LB Than Rau Re Loi Tirag b$ p h ^ trcD cSy b ^
Hinh 2: Ho£it tinh chdng oxy hda tdng v l khu s i t a ttmg bo phto trfin ely ngd
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La Than Rka M Loi Timg phan Iren ciy bip
Hmh 3: Ho?it tinh bat gdc tii do DPPH Of timg phan cly ngfi
Tir hinh 1 v l 2, thay ring htoi lupng clorophyl, boat tinh ehdng oxy hda tong vl khu sat d 11 cao hon so vdi cle bo phan khie eua cay, ung vdi 1,84 mg clorophyl/g mlu, 8,748 ± 0,003 g axit aseobic/lOO g mlu va 18,32 ± 0,001 g FeSO^/lOO g mlu. Trong khi dd DPPH cua la va rfi eay ngd lai tdt ban elc thtoh phln cdn lai tren eay, iing voi 72,6% va 74,8%. Trong khi dd loi ngo co kha ntog b i t gdc tu do thip nhat ling vdi 53,7% (Hinh 3). Kha nang bat gde tu do DPPH d tung phln eay ngd ed dp dao dong ldn vao khotog 20%.
Phto tieh ANOVA v l hdi quy thay rang, h i m luong polyphenol, clorophyl vdi boat tinh chdng oxy hda b timg phto doan ed su tuong quan manh me vdi nhau. Siic toh huong cua polyphenol ddi vdi boat tinh khli sit manh hon so vdi su toh huong ciia clorophyl. Tuy nhifin, su tuang quan giira ham luong clorophyl ddi vdi boat tinh chdng oxy hda tdng lai manh me ban so vdi hoat tinh khii s i t
So stoh vdi cac ket qua nghien ciiu tren the gidi thay ring: t r i xanh, t r i fi long v l tra den dupc chiet vdi nudc, sau do phto doan vdi ele dung mdi nhu:
clorofom, etyla axfitat v l butanol. Flavanol tdn tai d- phto doan etyla axfitat, trong khi do alkaloid trong phto doan clorofom. Thearubigin ton tai trong phto doan butanol, v l axti amin ehi tdn tai trong phto doan nudc (Xie va cs, 1993). Nhieu nghifin ciiu khae tren the gidi cung ehi ra boat tinh ehdng oxy hda v l him lupng polyphenol o cle loai thuc vat khic nhau (Xie vi cs, 1993; Namjooyan vi cs, 2010;Emilio vi cs, 2011). Ham lupng polyphenol, clorophyl d cly ngd cao hon 6 loai rau bae h i b Szczecin - Ba Lan (Monica vi cs, 2009) v l tuong duong vdi 18 loai thue vat o Brazin (Maria vi cs, 2010). Him lupng clorophyl d tao Spirulina platensis nudi tai Artoraissi v l Brandi Cong hoa Chad dat tuong iing 9,74 mg v l
7,26 m g / g chit khd, cao ban rat nhidu so vdi cay ngo (Ngakou vi cs, 2012).
Dieu do cd th^ thiy ring eay ngd eijng c6 hoEit tinh ehdng oxy hda va h i m luong clorophyl, polyphenol nhu cac thuc vat khie trfin the gidi. Dich chiet nly hoto toto co th^ hda rIn lam eae s t o pham gil tri gia ttog gilu boat chat polyphenol, clorophyl vdi boat tinh chdng oxy hda nhlm ntog eao siic khoe cho con ngudi.
3.2. H i m lupng potyphenol, clorophyl vdi hogt tinh ehdng oxy hda b ttmg p h t o doan dung mdi
3.2.1. Phin doan n-hexan
Dieh chidt ethanol ddi vdi ca cay ngo sau khi dupe cd dac, chung dupc tien htoh phto doto b&ig n-hexan theo cic t^ lfi dung mdi n-hexan {P^f dich ethanol cd dac (B) nhu sau: 1/1, 2 / 1 , 3/1 vl 10/1, Ket q u i thu dupc a trong phto doan n-hexan d polyphenol, clorophyl, Upit v l mpt sd sic td khac, Didu nly ed the n h t o dinh polyphenol hda tan nay tdn tai d" trang thli khdng phln cue. H i m lupng hpit cue thap, chi bing 1/15 lln so vdi h i m lupng ciia polyphenol.
Hinh 4: H i m lupng polyphenol, clorophyl trong tung p h t o doto n-hexan
B Ho}l dnh ctnng oty hoa iSng (m^mll
• H o ^ l U k h u s i l l i D g ^
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Hinh 5: Host tinh chSng oxy h6a tong v4 Idlft s5t trong timg plian d o ^ n-hexan Ham luong polyphenol, clorophyl voi hoat tioh chong oxy hoa khi phan doan voi ty 1^ li hexan/ethanol la 1/1 luon cao hon so vol cac ty t khac, tmmg ling voi 1,214 ± 0, 012 pg dorophyl/ul va 0,352 ± 0,011 mg axit galic/ml. Clorophyl chilli
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 12/2014
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chiet hoan toto khoi dich ethanol cd dae khi su dung den ty lfi 10/1 (n-hexan/ethanol), m^c dii truac dd da su dung cle ty le 1/1, 1/2 v l 1/3 (ethanol/n-hexan) de p h t o doan. Htoi lupng polyphenol thu dupc sau khi phto doan bang n-hexan tuong iing 12,9% so vdi tdng sd polyphenol tix dieh ethanol cd dac. Di^u nay hoto toto phil hpp vdi elc ket q u i nghifin euu trudc day ve clorophyl.
Hinh 6: Ho^t tinh b i t gdc tu do DPPH tt-ong tiing phto d o ^ n-hexan
Nghifin ciiu ve boat tinh ehdng oxy hoa cua phto doan n-hexan, thay rang str dung ty le n- hexan/ethanol cd dae 1/1 la eao nhat. Ho^t tinh ehdng oxy hda tdng va khu sit 6 p h t o doan 1/1 chiem hon 80% so vdi tong 3 phto doan d, 1/3 v l 1/10. Hoat tinh bat gde tu do DPPH cao nhit d phto doan 1/1 khotog 78% v l thap nhat d phto doan 1/10 11 khoing 68%. P h t o tich ANOVA v l thdng kfi thay ring su tuong quan duong giua h i m luong clorophyl, polyphenol voi boat tinh ehdng oxy hoa II kha cao (R^ > 0,8). Su tuong t i e qua lai giua cic ham mue tifiu tuto theo mo hinh phi tuydn logarit tiem can true holnh. Tliue nghifim eho thay d^
:hiet hoan toln clorophyl khoi dich ethanol cd dac
; l n lupng n-hexan gip 16 lan so vdi lupng dich 3thanol da cd d | c . Ket qua nghifin ciiu ve ho^it tinh
;hdng oxy hda d p h t o doan nay hoto toan phii hpp fbi cac ket q u i da cdng bd tren the gioi. Namjooyan /a es (2010) da ehi ra h i m lupng phenoHc tong rong phto doan n-hexan dat 0,014 mg axit jahe/lOOg. Hoat tinh khii sit v l b i t gdc tu do )PPH tirong irng 1,305 mmol Fe^Vl va 5,2 mg/ml.
imilio v l es (2011) eiing su dung n-hexan M. phto l p clorophyl.
Nhu v|y, phto doan n-hexan hoto toto phii hop rong vific tinh chd clorophyl boat tinh sinh hoc iing lung trong thuc, dupe pham. Dly se 11 phto doan iem n t o g mang tinh dOt pha ve sle td clorophyl o
^fit Nam cung nhu trfin the gioi.
3.2.2. Phin doan ethanol
Sau khi thu dupc phto doan n-hexan, tiep tuc Ilm saeh clorophyl va polyphenol b i n g each phto doan hang ethanol 96%. Ket qua eho thay su tuong ddng vd h i m luong clorophyl, polyphenol va hoat tinh d c i c phto doan theo ty le khic nhau MiA khi phto doan bang n-hexan. Him lupng clorophyl dupre chiet sang phto doan ethanol chi chidm 46% so voi tdng sd clorophyl trong dieh ethanol co dac. Him lupng polyphenol khong phto cue duac Itoi sach bing ethanol chi ehiem 46% so vdi tong polyphenol 6 phln doan n-hexan. Kdt qua cho thiy, him luong polyphenol cao nhit d' phto doan Ll 1:1 tuong iing vdi 0,316 ± 0,003 mg/ml. Tai phto doan Ll nly, h i m lupng clorophyl e t o g cao nhat, iing vdi 0,559 ± 0,002 mcg/ml (hinh?).
8 Hdm hiijng chlorophyll (mcg/ml) Q H ^ lupng polyphenol (rag/ml)
Hinh 7: Him lupng polyphenol trong timg p h t o doto ethanol
Hinh 8: H i m lupng clorophyl trong ttoig phto doto ethanol
Hinh 9: Hoat tinh chdng oxy hda tdng v l khu sit trong tung phto doto ethanol
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Hoat tinh chdng oxy hoa tdng va Idiu sit d phto doto nly k h i manh, tuong iing vdi 2,468 mg axit ascobie/ml v l 6,021 mg FeSOi/ml. Hoat tinh bat gdc hi do DPPH dat 66,75 - 76,25 (%) (hinh 9) b tting phto doto ti-ong phto doan ethanol nly. Dinh gil su tuong quan giiia cle ham mue tifiu va phto tieh ANOVA thay ring, chiing tuong tac manh me vol nhau theo h i m phi tuydn.
Cac nghifin ciiu tren the gidi eho thiy, phto doan ethanol tach tir phan hoa Bee Venezuelan cd h i m luong polyphenol vdi hoat tinh chdng oxy hda eao hon so vdi phto doan mehanola v l nuac {Perez- Perez vi cs, 2012). H i m Iupng polyphenol trong phto doan dao dOng trong khotog 396,7 -1286,7 mg gahc/lOOg phln hoa; hoat tinh ehdng oxy hda tuong duong 0,50 1,84 micromoles Trolox TEAC/lOOg.
Nhu v | y phto doan etiianol nly da lam sach dupc phan polyphenol khdng phto cue v l tich dupc mpt phln boat chat cd dp phto cue thap vdi boat tinh nhit dinh. Vdi htoi Iuong v l hoat tinh nhu da trinh biy, phan doan nly hoto toto cd the iing dung trong di^u ch^ thue pham gilu polyphenol boat tinh.
3.2.3. Phan doan etyla axetat
Hmh 10, 11 va 12 minh hpa chi tidt h i m lupng polyphenol, clorophyl v l boat tinh chdng oxy hda it phto doto- Him lupng polyphenol cao n h i t 6 phSn doan Ll 1:1 ttiong iing vdi 2,378 ± 0.16 mg/ml (hinh 10). Hoat tinh chdng oxy hda tdng va khu sit cQng eao nhat d phto doan Ll 1:1, tuong duong vdi 5,584
±0,1 mg axit aseobie/ml v l 12,131 t 0,13 rag F e S 0 4 / m l ( h m h l l ) .
12.131 ±0.13 a Ho?i liih chAng oxy h4a l4ng (n B Ho»I dnh Mm stt {mg/iaS)
2,378 ± 0
~h
6
1,222 ±0
fki
I I
0,732±0.15
i
0,423 ±0.11 han d o ^ L l Phan iSwn LlHmh 10: ffim lining polyphenol trong timg phan doan etyla axetat
Dich chiet ethanol con lai sau khi phan doan bang n-hexan, duoc a^p tuc phan doan bang etyla ax4tat Ket qua cho thay luong eiyla axetateetyla axetat can dung de lam sach polyphenol khoang 16/1 (etyla axStat/ ethanol). Ham luong clorophyl trong phan doan nay khong xac dinh th^y. Ham Iuong polyphenol dat 0,423, 0,732, 1,222, 2,378 mg axit gahc/<->ml tuong iing phan doan etyla axelat/ethanol 10/1; 3 / 1 ; 2 / 1 va 1/1. Hoat tinh chong oxy hoa tong va khu s i t ciia tong c4c phto doan trong phan doan etyla axetat nay toong ling 11,634 mg axit ascobic/ml va 26,202 mg FeSOj/ml.
PhSn dotn .thanol/ n.hei Hinh 11: Hoat hnh ch6ng oxy h6a trong timg phan
doan etyla axetateetyla axetat Nihal v£i cs, 2007 chi ra ham lu<;ng polyphenol trong la t r i tuoi vi t r i xanh (tuong duong 680,2 vJ 560,8 mg axit galic/g dich chi&). Phin doan elyla axetat cua dich chift nudc tir 14 t r i tuoi v i t r i xanl CO hoat tinh chdng oxy h6a (80,8 va 78,6 g axil ascobic/ lOOg dich chi^t). Hd so tuong quan giua ham luong polyphenol v i hoat tinh chdng oxy hda r^t manh (R^ = 0,9376 doi voi 14 t r i tuoi v i 0,9783 ddi voi t r i xanh). Ham luong polyphenol tuong ling 0,1748 ± 0,005 g phloroglucinol/g ch& khd. Hoat tinh chdng oxy hoa tdng v i idiu s i t toong irng 0,1812 ± 0,004 g axit ascobic/g chat khd v i 0,8866 ± 0,0023 g FeSOj/g chat khd. Hon nira, Cho vi cs (2012) chi u h i m luong polyphenol cua rong nau Ecklonia cavi thu d phan doan etyla axdtat la 367,42 m g / g vdi 5 loa polyphenol khic nhau (phloroglucinol, eckstolonol eckol, triphlorethol-A v i dieckol). Polyphenol thu ti phin doan niy ciing cao hon so vdi cac phan d o ^ n hexan, n-butanol v i nudc (Cho va cdng sir, 2012).
Hinh 12: Ho?t tinh bSt gdc tl; do DPPH trong timg phin dosn etyla axetat
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Hoat tinh h i t gdc ty do DPPH o phto doan nay tirang iing 61,79%, 63,81%. 70,37% v l 76,51% ddi voi tiing ty le etyla axetat/ ethanol 10/1; 3/1; 2/1 v l 1/1 (hinh 12). Phto tich ANOVA va hdi quy thay rtog h i m lupng polyphenol vdi ho^t tinh ehdng oxy hda ttiong quan duong manh me vdi R^ lon hon 0,8 va tuto theo phuong trinh logant tifim can true holnh.
Sanaa M. va cs (2011) cung chi ra, rong luc Enteromotpha compressa (linnaeus) Nees, Ulva lactuca, v l E. linza, dupc thu d vinh Abu Qir d Alexandria (Mediterranean Sea) dupe phto doan vdi elc dung mdi ed dp phto cue ttog din (petroleum fite, clorofom, etyla axfitat v l nudc). Phto do$n etyla axfitat c6 kha ntog b i t gdc tii do DPPH (82,80%) (Sanaa vi cs, 2011). T i t ca cic nghien ciiu deu chi ra h i m lupng polyphenol vdi boat tinh chdng oxy hda chidm ^ le phan trtoi eao trong phto doan etyla axfitat thu dupc, ddng nghia vdi su xae dinh polyphenol chdng oxy hda thu dupe tti phto doan etyla axfitat ed dd sach ban so vdi ele phto doan khic.
Tu do thiy ring, phto dogn nly 11 mpt trong nhung phto doto quan trpng trong vific tmh saeh polyphenol phto eue cd ngudn gde tti cay ngd. Phto doan etyla axfitat nly phu hpp eho vific dieu che polyphenol, clorophyl boat tinh trong y dupc.
3.2.4. Do sach cua polyphenol vi clorophyl qua timg phin doan
Ket qua nghifin ciiu cho thay dich chiet ban diu 6 didu kifin nghifin ciiu duoc phto Ilm bdn phto doto khie nhau: n-hexan, ethanol v l etyla axfitat Him lupng polyphenol trong phto doan n-hexan chidm 32,6% trfin tdng sd chat cd trong phto doan.
Nhu vay dp sach eua polyphenol da tang Ifin 20%.
H i m lupng clorophyl thu dupc tuong iing 0,3% chat khd.
Dich ehiet sau khi dupc phto doan bing n-hexan tien htoh phto doan bing ethanol. Phto doan nly h i m lupng polyphenol chi^m 42,4% chit khd thu dupc. Nhu vay dd sach cua polyphenol da ttog len 9,8% so vdi phto doto n-hexan. Dp sach cua polyphenol tai phto doan nay cung eao hon so vdi phto doto n-hexan. So stoh vdi phto doto n-hexan thiy ring, phto doan ethanol nly ed him lupng polyphenol va boat tinh ehdng oxy hda manh hon.
Dich chiet con lai sau khi phto doan btog n- hexan ti^n htoh phto d o ^ bang etyla-^fitat Phln
tich so stoh vdi cac ket q u i trudc v l sau phto doan nay thay polyphenol 36,8%, ddng nghia vdi dp sach eua polyphenol ttog len 24,2% so vdi ban diu. P h t o doan etyla axfitat eho ham luong polyphenol v l hoat tinh chdng oxy hda la cao nhat tuong iing 4,755 mg/ml, 11,634 mg axit ascobic/ml v l 26,202 mg FeSOi/ml. P h t o doan nly cho thiy eo dp sach cao nfin tidp tuc chay qua cot Sephadex LH 20 dd x l e dinh dp sach.
Dich ehiet con lai sau khi phto doan bing etyla axfitat 11 phto doan ethanol con lgi. Phto dogn ethanol con 1^ nly chua 24% polyphenol so vdi tdng polyphenol ban diu. Ddng nghia vdi dp sgch eua polyphenol ttog len g i n 10% so vdi ban diu. Phan doan ethanol sau n-hexan saeh hon so vdi phto doan n-hexan va etyla axfitat Nhu vay thay ring cac phto doan ethanol, n-hexan v l etyla axfitat cd the iing dung trong dieu che che pham polyphenol, clorophyl ed dd sach cao ung dung trong thuc dupe phim.
4. KFTLUAN
Him lupng polyphenol o thto cay ngd 11 nhi^u n h i t ung voi 2,41 ± 0,001 g axit galic/lOOg mlu khd.
Him lupng clorophyl vdi hoat tinh chdng oxy hda tong vl khii sit d ' n eay ngd cao nhit iing vdi 1,84 mg clorophyl/g m l u kh6 vdi 8,748 ± 0,036 g axit ascobic/ lOOg m l u va 18,32 ± 0,002 g F e S 0 4 / lOOg mlu. DPPH ciia II v l rfi cay ngo tdt nhat tuong iing 72,6% v l 74,8%.
Phln doan etyla axetat cho h i m luong polyphenol va boat tinh ehdng oxy hda la cao n h i t O phto doan etyla axfitat nly, h i m luong polyphenol dat 368 m g / g chat khd v l dp sach cua polyphenol ttog Ifin 24,2% so vdi ban diu. Hoat tinh ehdng oxy hda tdng, khii sit vdi bat gdc tti do DPPH tuong duong vdi 11,634 mg axit ascobic/ml v l 26,202 mg F e S O / ml vdi DPPH cao nhat dat 70,37%.
Phan doan n-hexan v l ethanol hoto toto ung dung trong dieu ehd polyphenol, clorophyl vdi boat tinh chdng oxy hda iing dung trong thtrc pham v l dupe pham.
PoljT)henol, clorophyl chidt tir cay ngd hoto toto ed the iing dung trong thue pham chiic ntog, y dupc, eung nhu ele linh vuc k h i c trong eude sdng. Cay ngo 11 ngudn cung cap phong phii v l da dang eae hoat chit polyphenol, clorophyl ed hoat tinh chdng oxy hoa phuc vu thuc pham, thuc pham chuic n t o g v l di^pc pham.
KHOA H O C C d N G N G H l
TMUEUDMMKIUO
1. Blois M. S., 1958. Antioxidant determinations by the use of a stable free radical. Natore, 26, 1199- 1200.
2. Cho Suengmok, Yang Hyejin, Jeon You-Jin, Justn Lee C , Jin Young-Ho, Back Nam-In, IQm Dongsoo, Kang Sung-Myung, Yoon Minseok, Yong Heim, Shimizu Makoto, Han Daeseok, 2012.
Phlorotannins of the edible brown seaweed Ecklonia cava Kjelknan induce sleep via positive allosteric modulation of gamma-aminobutyric acid type A- benzodiazepine receptor: A novel neurological activity of seaweed polyphenols. Food Chemistry, 132,1133-1142.
3. Emilio Alvarez-Pairilla, Laura A De La Rosa, Ryszard Amarowicz, and Fereidoon Shahidi, 2011.
Antioxidant Activity of Fresh and Processed Jalapeflo and Serrano Peppers. J. Agric. Food Chem., 59,163- 173.
4. http://vaas.vn/ldenthuc/cayngo/Monanton golathuocchuabenh.php.
5. Levent Inanf. A , 2011. Chlorophyll:
Structural Properties, Health Benefits and Its Occurrence in Vugin Olive Oils. Akademik Gida, 9(2), 26-32.
6. Lichtenthaler K. Hartmut and Buschmann Claus, 2001. Current Protocols in Food Analytical Chemistiy. F4.3.1 - F4.3.8.
7. Maria do Socorro M. Rufino, Ricardo E.
Alves, Edy S. de Brito, Jara Perez-Jimenez, Fulgencio Saura-Calixto, Jorge Mancini-Filho, 2010. Bioactive compounds and antioxidant capacities of 18 non- traditional tropical fruits from Brazil. Food Chemistiy, 121, 996-1002.
8. Monika Grzeszczuk, Dorota Jadczak, 2009.
Estimation of biological value of some species of mint (MentiiaL). Herbapolonica, 55(3), 193 -199.
9. Namjooyan F., Azemi M. E., IMimanian V.
R., 2010. Investigation of antioxidant activity and total phenolic content of various fractions of aerial parts of Pimpmella barbata (dc.) Boiss. Jundishapur Joumal of Natoral Pharmaceutical Products, 5(1), 1-5.
10. Nihal Turkmen Erol, Ferda Sari, Gdkfe Polat, Y. Sedat Velioglu, 2009. Antioxidant and Antibacterial Activities of Various Extracts and
Fractions of Fresh Tea Leaves and Green Tea. Tariin
BilimleriDergisi, 15(4), 371-378. ^ j | 11. Ngakou Albert, Ridine Wague, MbaiguinamJ^I
Mbailao, Namba Fabienne, 2012. Changes in t™'*
physico-chemical properties of Spirulina plateni from three production sites in Chad. Joumal of Animal & Plant Sciences, 13(3), 1811-1822.
12. Perez-Perez E. M., Vit P., Rivas E., Sciortini) R, Sosa A , Tejada D., Rodriguez-Malaver A J., 2012.
Antioxidant activity of four color fractions of bee pollen from Merida, Venezuela. Arch Latinoam Nutr., 62(4), 375-80.
13. Prieto P., Pineda M. and Aguilar M., 1999, Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Analytical Biochemistiy, 269,337-341.
14. Rebecca J. Garlock, Shishfr P. S. Chundawat Venkatesh Balan and Brace E. Dale, 2009, Optimizing harvest of com stover fractions based on overall sugar yields following ammonia flba expansion pretreabnent and enzymatic hydrolysis Biotechnology for Biotoels, 2(29), 1 - 14.
15. Stefano Petti, Crispian Scully, 2009.
Polyphenols, oral health and disease: A review.
Joumal of dentishy, 37,413 - 423.
16. Samuel Abiven, Alexander Heim, Michael W, L Schmidt, 2011. Lignin content and chemical characteristics in maize and wheat vary between plant organs and growth stages: consequences for assessing lignin dynamics in soil Journal of Plant and Soil, 343(1-2), 369-378.
17. Sanaa M. M. Shanab, Emad A Shalaby, and Eman A. El-Fayoumy, 2011. Enteromorplil compressa Exhibits Potent Antioxidant AcUvilJ.
Journal of Biomedicine and Biotechnology, 11 pages.
18. Swanson A K. and Draehl L D., 2002, Induction, exudation and the UV protective role of kelpphlorotannins. Aquatic Botany, 73,241-253.
19. Trang tam Tin hoc va Thdng ke - Bo Ndng nghi«p v i Phit blen Ndng thon, 2013. Bio cio kS qua thuc hidn ke hoach 7 thing nam 2013 ngank ndng nghiep v i phit trien ndng thdn.
66 NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 12/2014
KHOA HOC C O N G N G H |
20. Xie B., Shi H., Chen Q., Ho C. T.. 1993. 21. Zhu Q. T., Haekraan R. M., Ensunsa J. L , Antioxidant properties of fractions and polyphenol Holt R. R and Keen C. L, 2002. Antioxidative constituents from green, oolong and black teas. Proc. activities of oolong tea. Joumal of Agricultural and Natl. Sci. Counc. Repub. Chma B., 17(2), 77-84. Food Chemistiy, 50, 6929-6934.
EVALUATION OF POLYPHENOL, CHLOROPHYLL CONTENT W i m ANTIOXIDANT A C I l V m E S OF EXTRACTION FRACTION FROM EACH PART OF MAIZE
Dang Xuan Cuong, Dao Trong Hieu Summary
This paper focused on researching about purification step for polyphenol, chlorophyll with antioxidant activities which original from maize, n-hexan, ethanol, ethyl acetate were used as purification fraction solvent. Polyphenol, chlorophyll content with antioxidant activities (total, reducmg power and DPPH) of each part of maize were evaluated. The results showed, polyphenol content of maize stalk was the most, chlorophyll content with antioxidant activities (total, reducing power) of maize leaves were the highest DPPH of maize leaves and roots M were the best. Polyphenol with antioxidant activities of ethyl acetate fraction were the highest.
Keywords: Polyphenol chlorophyll, antioxidant, maize, 6-action.
Ngudi phln b i ^ : TS. Trin Thi Mai Ngly nhin bli: 1/7/2014 Ngly thdng qua phln bi$n: 1/8/2014 Ngly duy^t dtog: 8/8/2014
