HAM LU'P'NG POLYPHENOL VA KHA NANG CHONG OXY HOA CUA 28 THU'C VAT AN DU'OC TAI VIET NAM
T r u o n g Tuy§t Mai, T r u a n g Hoang Kien*, Nguyin Cong KhSn, Nguyen Van Chuyen Vien Dinh duffng Quoc gia
'^ Chi cue ve sinh an todn thuc phdm Qudng Ninh
"Khoa Dinh duffng vd Thuc phdm, tru&ng Dgi hgc Phu nd- Nhdt Bdn
T O M T A T :/v;...
Kha nang chdng oxy hda cua 28 thye vat an dupc ciia Viet Nam, du'di dang nudc va dang methanol, gdm 4 nhom (thu'c vat dung lam dd udng, rau dai an dugc, rau gia vj, va rau la xlim mau), da dugc danh gia tren phdng thi nghiem. Kit qua, nhom thye vat dung lam dd udng the hien hoat tinh cao nhdt vd kha nang ch6ng oxy hda, tidp theo la ddn nhdm rau dai an dugc, nhdm rau gia vi va cudi cung la nhdm rau sdm mau. Mdi quan he thuan chilu va duang tir:i gida kha nang chdng oxy hda vdi ham lu'gng polyphenol cua 28 thye vat da du'pc xac dinh. Bdn nguyen lieu tidm nang co hoat tinh sinh hpc bang hoac tdt han la oi, bao gdm la Tram Vo do, Nu Vdi, la Sang mau va la Vdng qua xoan, da chi ra kha nang ch6ng oxy hda cao (tu'ang dng 85,87, 78 va 80) tai ndng dp 30 mg nguyen lieu dong kho/ml dung dich. Cac nguyen lieu nay cung co ham lu'gng polyphenol eao mpt each dang ke (tu'ang ling vdi 251.7, 146.6, 136.6 va 168.6 mg catechin/g nguyen lieu dong khd). Chinh vi vay, 4 nguyen lieu nay co the du'oc xem la ngudn nguyen lieu mdi tilm nang v l kha nang ehdng oxy hda, thich hgp ddi vdi viee sd dung nhu" cac thye pham chdc nang trong tuang lai.
TCf khoa: ehdng oxy hda, ham lugng polyphenols, thye vat an dugc Viet Nam
L DAT VANDE
Tii cac nghien ciiu dich te hgc va cac nghien ciiu thii nghiem da cho t h a y tieu thu cac do udng va thiic an giau polyphenol cd mdi quan he cd y nghia vdi viec giam cac nguy cd sinh benh man tinh khdng lay, trong dd cd lienh dai thao dudng [1,2J.
Polyphenols trong t h u c v a t dugc xem la thanh phan quan trgng ddi vdi k h a u phan an ciia con ngudi. Cac t h a n h p h a n nay cd rat nhieu cac hieu qua boat tinh sinh hoc, bao gdm kha nang chdng oxy hda, k h a nang chdng tc bao ung thu...
Trong cac nghien ciiu tren phdng thi nghiem cho thay, t h a n h p h a n polyphenol trong thuc vat c6 kha n a n g giam oxy hda stress thdng qua viec triet tieu cac phan tii oxy gem hoat tinh va k h a n a n g lam giam
peroxyt hda lipid. Tren ddng vat, cac n h a nghien ciiu ciing da chi ra vai trd bao ve ton thuong te bao thdng qua kha n a n g chdng oxy hda ciia polyphenol, hd trg giam cac bien chiing ciia mgt so benh lien q u a n den t a n g oxy hda, n h u benh dai t h a o dudng, benh tim mach...[1,2].
Viet N a m la mgt nudc nhiet ddi, da d a n g cac loai thuc vat, cd nhieu cac bai thudc dan gian sii dung thuc vat de hd trg viec phdng va tri benh. H a m lugng va chat lugng polyphenol va boat tinh sinh hgc ciia t h u c vat vimg nhiet ddi dugc xem la td't hon ca do su tiep xiic nhieu vdi anh sang m a t trdi.
Trong khi dd, viec nghien ciiu d a n h gia t h a n h p h a n polyphenol va hoat tinh cac thuc vat d Viet N a m cdn rat it [3]. Can phai tien h a n h nghien ciin sang lgc tren cac thuc Tacgia: Truong Tuyet Mai "'
Dia chi: Vien I )inh dudng, 48 B Tang Bat Hd Dien thoai: 09 I 9911 777; 04-39716058 Email: [email protected]
Tap chi Y hpc du phong, Tap XX, sd 2 (110) 57
vat an dugc nhdm tim ra cac nguyen lieu tiem n a n g giau polyphenol va boat tinh sinh hpc cao cd tac dung hd trg phong va tri benh m a m t i n h khdng lay. Chinh vi vay, chiing toi tien h a n h thuc hien nghien ciiu vdi muc dich d a n h gia h a m lugng polyphe- nol va k h a n a n g chdng oxy hda ciia 28 thuc vat an dugc tai Viet Nam.
I L P H U ^ O N G P H A P N G H I E N C l T U 2.1. Nguyen lieu va chi§t tach.
28 loai thuc vat an dugc ciia Viet Nam dugc m u a tii cac chg dia phuang tai ca mien Bdc va mien Nam, Viet Nam. Viec xac dinh ten khoa hgc, ten Latinh ciia 28 loai thuc vat n a y d u a tren cudn sach
"Nhiing cay thudc va vi thud'c Viet N a m "
cua Dd Ta't Lgi [4] (bang 1). Cac nguyen lieu dugc chia t h a n h 4 nhdm, nhdm thuc vat ddng de lam do udng (5 loai), nhdm r a u dai an dugc (11 loai), nhdm r a u gia vi (5 loai), va nhdm r a u xanh thdm (7 loai). T a t ca cac nguyen lieu dugc lay phan la banh te, rieng chi cd Vdi va Hde la lay phan nu.
Sau khi t h u hoach, cac nguyen lieu thd dugc riia sach, lam ddng khd, sau dd xay t h a n h bgt min va bao quan tai nhiet do - 30°C cho den khi sii dung.
Su d u n g 2 dung mdi la nudc va methanol 50% de chiet tach, vdi ty le 100 mg bgt nguyen lieu khd/5 ml dung mdi, dun tai nhiet do 100°C, ly tam va thu dung dich de lam thi nghiem.
2.2. Xac dinh h a m lu'Ong polyphenol Ham lugng polyphenol dugc xac dinh theo phuong p h a p so m a u ciia F o l i n - C i o - calteu [3], do tai budc sdng 756 nm. Cac ke't qua dugc dien t a tuong duong vdi m i l - ligrams ciia catechin t r e n 1 gram nguyen lieu kho (mg catechin/g dw). Mdi mau dugc thuc hien 3 lan trong dieu kien tuong duong.
2.3. Phuo'ng p h a p xac djnh kha nang chong oxy hoa.
C h u a n hi dung dich ddng n h a t nao chudt. Thuc hien lay n a e cac con chupt khde m a n h , sau dd ddng n h a t nap chudt bang dung dich dem (0.1 - phosphate tai
pH 7.4) vdi ty le IC^r. Sau khi ly t a m , d u n g dich t h u dugc bao quan tai - 80°C cho den khi sii dung. J / ' . - iJ
De xac dinh k h a n a n g iic che su p e r - oxy hda lipid tren cac liposomes nao chugt, tien h a n h cac budc thi nghiem theo tac gia Ohkawa va cdng su [5 ] vdi mgt sd t h a y ddi nhd. Mdi d u n g dich m a u chiet tach dugc pha loang vdi nudc cat hoac methanol 50%
tai cac ndng dg khac n h a u . H e n h g p b a n d a u bao gdm 30 j^L dung dich m a u chiet tach, 45 |iL N a H C 0 3 0.2 M, 150 pL a s c o r - bic acid 0.05 mM va 975 |iL d u n g dich dem phosphate (0.1 M, pH 7.4), hdn hgp dugc li am trong 10 phiit tai 37°C. S a u khi t h e m 300 iiL ciia dung dich ddng nha't nao chudt, p h a n ling tiep tuc xay ra trong 60 phiit tai 37°C. Bd s u n g them 1500 |aL d u n g dich thiobarbituric acid 0.5% vdi T C A - H C l (20% w/v trichloroacetic acid trong 0.2 N HCl) va giu tai nhiet do sdi 100°C trong 30 min. Lam lanh, ly tam, dung dich t h u dugc do tai budc sdng 532 nm. T h a n h p h a n C a t - echin dugc sii dung la t h a n h p h a n t h a m khao. Mdi m a u se thuc hien 3 lan va xac dinh ty le lie che su peroxy hda lipid dugc xac dinh bang each so sanh vdi ong chiing.
Ndng dd cudi ciing ciia m a u chiet tach dugc xac dinh dudi dieu kien thi nghiem va dugc dien ta bang microgram nguyen lieu ddng khd t r e n lyophilized millilit dung dich. Gia tri IC5Q la ndng do cudi ciing ciia m a u trong hdn hgp p h a n iing m a dugc xac dinh n h u lugng polyphenol (tuong duong mmol c a t - echin) can thiet de lie che 50% s u peroxy hda lipid dudi dieu kien thi nghiem.
2.4. P h a n tich thong ke.
Ta't ca gia tri IC50 dugc tinh toan tii dudng cong lieu dap iing. Cac m i u chiet tach cd boat ddng iic che dudi 40% tai lieu dau tien ciia cac m a u chiet tach p h a loang se khdng dugc xac dinh IC50. T a t ca cac ket q u a dugc dien ta bang gia tri t r u n g binh ciia 3 thii nghiem khac n h a u . P h a n tich mdi q u a n he thdng ke su dung p h a n mem S P S S version 12.0. Gia tri p<0,05 dugc xem la mdi quan he cd y nghia thdng ke.
I I I . K E T Q U A V A B A N L U A N 58 Tap chi Y hgc du phong. Tap XX, sd 2 (110)
Ham lu'ong polyphenol
Bang 1. Ham lu'gng polyphenol va hoat dgng chong oxy hoa cua chiet tach dung djch nirac va methanol tir 28 thiPC vat an difcc a Viet Nam.
Nguyen lieu thiPc vat
Ten khoa hoc Ten Viet Nam
Ham
Nuf&c
lu'O'ng Polyphenol (mg/g dvi)'^
Methanol
NU'O'C (%) In.^
Hoat dong chong oxy hoa^
ICsof
Methanol (%) In.^ IC50' Nhom thirc vat diing uong
Psidium guajava*
Cleistocalyx operculatus'' Sophora japonica'' Camellia sinensis^
Nelumbo nucifera^
(Trung binh)
Oi
Vol Hoe Che Xanh Sen
122.8 122.5 105.5 84.8 83.5 103.8
169.7 146.6 165.4 111.9 133.5 145.4
75 87 56 83 74 75
3.02 1.34 5.40 1.12 3.80 2.94
80 87 65 84 76 78
2.81 1.12 5.83 1.06 3.41 2.85 Nhom rau dai an dvorc
Syzygium zeylanicum^
Horsfieldia amygdalina^
Careya arborea^
Artemisia vulgaris^
Ficus racemosa^
Houttuynia cordata^
Spondias pinnata^
Polyscias fruticosa^
Paederia lanuginosa^
Premna corymbosa^
Centella asialica^
(Trung binh)
Tram Vo do Sang mau VCrng Qua Xoan Ngai cdu Sung Diep ca Coc Chua Dinh Lang Mo long Vong Cach Rau Ma
176.9 91.5 95.3 49.7 41.2 40.8 32.9 29.2 28.8 25.5 24.3 57.8
251.7 136.6 168.6 64.2 87.2 56.1 40.8 34.0 32.2 43.5 31.5 86.1
84 75 80 43 39 14 16 18 2 38 27 40
2.12 3.81 4.23 8.80 9.65 41,86 39.60 48.63 ND 6.22 9.19 17.41
85 78 80 58 68 29 12 16 4 63 36 48
1.71 3.45 4.09 6.90 5.73 35.21 34.83 42.47 ND 3.68 7.43 14.55 Nhom rau gia vj
Enydra fluctuans^
Perilla frutescens^
Polygonum odoratum^
Eryngium foetidum^
Ocimum basilicum^
(Trung binh)
Rau Ngo Tia to Rau Ram Mui Tau Hung Que
55.9 50.3 32.5 19.1 12.9 34.1
80.6 66.8 64.8 22.5 19.2 50.8
56 36 32 4 8 27
3.63 7.85 6.23 ND 12.45 7.54
60 39 40 5 24 37
2.83 7.42 7.85 ND 11.84 7.49 Nhom rau xanh tham
Ipomoea batatas^
Sauropus androgynus^
Nasturtium officinale^
Blumea lanceolaria^
Piper lolot^
Ipomoea aquatica^
Cucurbita pepo^
(Trung binh)
Thanh phan tham khao Catechin
Khoai Lang Rau Ngot Cai Soong Xircng Song La lot Rau Muong B i N g o
68.4 29.3 25.4 24.6 19.6 18.5 17.2 29.0
83.6 28.4 27.4 25.5 20.9 26.9 19.7 32.2
68 0 11 32 0 16 2 18
4.05 ND 23.61 18.28 ND 26.24 ND 18.05
69 4 12 37 0 31 0 22 50 ng/ml 29
3.22 ND 30.64 17.22 ND 18.25 ND 17.33 nmol 58.91
"sir dung phdm Id. ^sir dung phdn nu hoa. ^&'^Chiet tdch 100 mf nguyen lieu kh6/5 ml nwac hodc 50%MeOH: Icet qud Id trung hinh cua 3 so. 'Dien Id tuang duang milligram catechin / lg mdu kho. ^IJc che (In.) lgi nong do 30 /jg nguyen lieu kho/ml dung dich (50 fjg/ml doi vai catechin) . 'Nong do cuoi ciing (/umol catechin) cdn thiit de ire che 50% sir peroxy hoa lipid duoi dieu kien Ihi nghiem. (Trung binh) diem ta Ining binh cua cdc gid tri moi nhom. ND: khong xdc dinh.
Trong bang 1 cho tha'y, nhin chung h a m lugng polyphenol trong dung dich chiet tach methanol cao hon dung dich chiet tach nudc. Ham lugng polyphenols cd su dao ddng dang ke trong 28 m a u t h u c
vat, h a m lugng tii 12.9 tdi 176.9 mg c a t e - chin/g nguyen lieu khd (dw) dd'i vdi d u n g dich chiet tach b a n g n u d c va. tii 19.2 tdi 251.7 mg/g ddi vdi d u n g dich chiet tach bang methanol. Trong 4 nhdm, h a m lugng Tap chi Y hoc du phong. Tap XX, sd 2 (110) 59
polyphenol cao nha't dugc quan sat thay d n h d m t h u c v a t diing de udng, vdi gia tri t r u n g binh la 103.8 mg/g (trong dung dich nudc) va 145.4 mg/g (trong dung dich methanol), tiep theo la nhdm rau dai an dugc (tuong duong vdi chiet tach nudc va chiet tach methanol: 57.8 va 86.1 mg/g), tiep theo la nhdm rau gia vi (34.1 va 50.8 mg/g), va cudi cung la nhdm rau xanh thdm (29.0 va 33.2 mg/g). Trong s d 2 8 mdu thuc vat, h a m lugng polyphenols cao nha't tim thay trong la Tram Vd dd (tuong duong vdi 251.7 mg catechin/g), la Viing Qua Xoan (168.6 mg catechin/g) va la Sang m a u (136.6 mg catechin/g) ddi vdi nhdm r a u dai an dugc, tiep theo la 3 loai ciia nhdm thuc vat dung de udng: la 6 i (169.7 mg c a t e - chin/g), nu Hoa hde (165.4 mg catechin/g), va n u Vdi (146.6 mg catechin/g).
Hoat dgng chong oxy hda
Bang 1 cho thay, trong 28 m a u thuc vat, cd 10 m a u thuc vat cd boat dgng chd'ng oxy hda cao (tai ndng do cudi ciing 30 \xg nguyen lieu khd/ml dung dich), lan lugt xep theo thii t u tii cao den t h a p : Nu Vdi > la Tram Vd Dd > la Che xanh> la Vimg Qua Xoan > la Oi > la Sang mau > la Sen > la Khoai lang > nu Hoa hde > la Rau Ngd.
De danh gia chat lugng boat ddng chd'ng
oxy hda ciia 28 m l u thuc vat nay, nghien ciiu da tien h a n h t i n h toan gia tri IC.50 (tuong duong vdi mmol catechin), vdi con sd cang cao cd y nghia chat lugng ciia viec chdng oxy hda cang t h a p . Nhdm t h u c vat dung ud'ng lai la nhdm cd k h a n a n g chdng oxy hda cao nhat, vdi gia tri t r u n g binh ciia IC50 la 2.94 |imol ddi vdi chiet tach bdng nudc va 2.85 |J.mol ddi vdi chiet tach bang methanol. Vi tri tiep theo thudc ve nhdm r a u gia vi, sau dd den n h d m r a u dai an dugc va cudng cimg la nhdm r a u xam mau.
So sanh chat lugng chd'ng oxy hda cua 28 m i u thuc vat bdng gia tri IC50 cho thay thii t u ciia 10 m a u t h u c vat ndi t r e n da cd su thay ddi ve vi tri tii cao den t h a p : la Che xanh > nu Vdi > la T r a m Vd do > la Oi > la Rau Ngd > la Sen > la Sang Mau > la Khoai lang > la Viing qua xoan > n u Hoa hde.
Moi quan he giira kha nang chong oxy hda va ham lugng polyphenol
Hinh IA va I B cho t h a y , b o a t ddng chdng oxy hda ciia 28 m a u thuc vat trong nghien ciiu nay cd mdi quan he t h u a n chieu vdi h a m lugng polyphenol. Mdi q u a n he duong tinh va cd y nghia thdng ke, r^ = 0.75 va p<0.01 ddi vdi dung dich nudc (hinh IA) va r2 = 0.69 va p < 0.01 ddi vdi dung dich methanol.
0 40 SO i 2 i 160 20<) 240 280 PoKphcnol contents (ma/g tin)
Hinh 1. Moi quan he giira hoat dgng c h o n g o x y hoa (antioxidant activities -%) va ham lu'O'ng polyphenol (polyphenol contents-mg catechin /g nguyen lieu kho (dw)) cua chiet tach n u a c (A) va
methanol (B) tir 28 t h g e vat an du'o'c cua Viet Nam.
60 Tap chi Y hpc du phdng, Tap XX, sd 2 (110)
Polyphenols thuc vat dang ngay cang dugc cac n h a khoa hgc quan tam ve cac lgi ich sinh hpc tiem nang cua chiing. Cac ket qua thi nghiem tren 28 mau thuc vat cho thay mau thuc vat mao eda ham lugng polyphe- nols cao ciing cd boat dgng chdng oxy hda cao, va dugc the hien bang mdi quan he duong t i n h va t h u a n chieu. Cac ket qua trong nghien ciiu nay giiip k h a n g dinh them vai trd cua t h a n h p h a n polyphenol trong viec chdng oxy hda ciia 28 thuc vat nay, ciing cimg vdi cac bao cao ciia nghien cuu trudc day (3, 6, 7) ma chi r a rang k h a nang boat ddng chdng oxy hda ciia mpt sd thuc vat phu thudc vao h a m lugng polyphenol cd trong chiing.
La Oi va la Che xanh da dugc mdt sd tac gia t r e n the gidi nghien ciiu ve h a m lugng polyphenol va tac d u n g chd'ng oxy hda ciia chiing da dugc chiing minh r a t rd, va trong nghien ciiu nay la Oi va la che Xanh cung van the hien la nguyen lieu cd h a m lugng polyphenol cao va k h a n a n g chd'ng oxy hda cao. Mgt trong ket qua chinh ma nghien cuu nay tim thay, dd la 4 nguyen lieu tiem n a n g mdi da dugc tim tha'y, dd la la Tram Vd dd, nu Vdi, la Sang mau va la Viing Qua xoan.. Theo n h u mieu ta ciia Giao su Dd Tat Lgi va mgt sd sach chuyen nganh thuc vat, ca 4 cay thuc vat ndi tren thupc cay t h a n go la xanh, mge d vung nhiet ddi nong am. Cay T r a m Vd Dd va cay Vdi thupc hg cay Sim (Myrtaceae family), cay Sang Mau thudc hg M y r i s t i - caceae va cay Vung Qua Xoan thudc hg Lecythidaceae. Theo kinh nghiem dan gian tai Viet Nam, nu Vdi thudng dung de n a u nudc ud'ng vdi muc dich giai nhiet, kich thich tieu hda, cdn cac la Tram Vd dd. Sang Mau, Viing Qua Xoan thudng diing de lam r a u an sdng va ciing cd tac dung chd'ng di ngoai. Va cho den nay, chua cd nghien ciiu
khoa hpc ve cac loi ich si nh hgc cua 4 thuc vat ndi tren.
IV. K E T L U A N
Cac ket qua tim thay trong nghien ciiu chi ra md'i quan he cd y nghia thdng ke giiia h a m lugng polyphenol va k h a nang chdng oxy hda ciia 28 t h u c vat an dugc d Viet Nam. Trong sd' dd, chi ra bd'n nguyen lieu mdi cd tiem n a n g ve chuc n a n g boat tinh sinh hgc trong viec hd trg phdng tri benh m a n tinh khdng lay lien cjuan den tang oxy hda.
(1)
(4)
Hinh 2. Bon nguyen lieu mo'i tiem nang (1) Tram Vd do, (2) Nu Vdi, (3) Sang Mau va (4)
Virng Qua Xoan.
T A I L I E U T H A M K H A O
1. Arts IC, Hollman PC. 2005. Polyphenols and disease risk in epidemiologic studies.
Am J Clin N u t r 81: 317-325.
2.Urquiaga I, Leighton F. 2000. P l a n t polyphenol antioxidants and oxidative stress. Biol Res 33: 55#64.
3. Thu NN, Sakurai C, Uto H, Van Chuyen N, Lien do TK, Yamamoto S, Ohmori R, Kondo K. 2004. The polyphenol content and antioxidant activities o f t he main e d - ible vegetables in n o r t h e r n Vietnam. J Tap chi Y hgc du phdng. Tap XX, sd 2 (110) 61
N u t r Sci Vitaminol 50: 203-210.
4. Dd Tat Lgi. 2001. Nhiing cay thudc va vi thudc Vict Nam. N h a xua't ban Y hpc, Ha Npi.
5. O h k a w a H, Ohishi N, Yagi K. 1979.
Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95: 351-358.
6. Cai Y, Luo Q, S u n M, Corke H. 2004.
Antioxidant activity and phenolic com- pounds of 112 traditional Chinese m e d i c i - nal plants associated with anticancer. Life Sci 74: 2157-2184. ' " :
7. Rice-Evans CA, Miller N J , Bolwell PG, Bramley PM, Pridham J B . 1995. The r e l - ative antioxidant activities of plant - d e - rived polyphenolic flavonoids. Free Radic Res 22: 3 7 5 - 3 8 3 .
THE ANTIOXIDANT ACTIVITY OF 28 VIETNAMESE EDIBLE PLANTS AND ITS RELATIONSHIPS WITH POLYHENOL CON- TENTS
;H
Truong Tuyet Mai, Truong Hoang Kien*, Nguyen Cong Khan, Nguyen Van Chuyen**
National Institute of Nutrition, Hanoi
* Quangninh Food Administration
** Department of Food and Nutrition, Japan Women's University
•)i-;J
The antioxidant activitiy of aqueous and methanolic extracts from 28 common Vietnamese edible plants, comprising 4 groups (plants used for making drinks, e d - ible wild vegetables, herbs, and dark green vegetables), were investigated in vitro. The polyphenol contents of these extracts were determined by the Folin-Ciocalteu method and calculated as catechin equivalents.
The extracts from plants used for making drinks showed the highest activities for antioxidants, followed by edible wild v e g - etables, herbs, and dark green vegetables.
Positive relationships between antioxidant activities and polyphenol contents of these 28 edible plants were found in both a q u e - ous and methanolic extracts. Four new
promising materials t h a t are similar to or better t h a n guava leaf extract, including Syzygium zeylanicum, Cleistocalyx o p e r - culatus, Horsfieldia amygdalina and Careya arborea demonstrated high a n - tioxidant activity (85,87,78 and 80, respec- tively) at the final concentration of 30 mg lyophilized material/ml solution. These four edible p l a n t s contained significant high polyphenol contents (equivalent to 251.7, 146.6, 136.6 and 168.6 mg of c a t e - chin/g dry weight, respectively). Thus, these four materials might be possible new sources of antioxidants suitable for use as functional foods in the future.
Key Words: antioxidant activity, polyphe- nols, Vietnamese edible plants
62 Tap chi Y hpc du phong. Tap XX, sd 2 (110)