Tieu ban Sinh hgc vd ngudn lgi sinh vgt Biin
SV TICH LUY VA PHAN BO PHLOROTANNIN CHONG OXY HOA TRONG MOT SO LOAI RONG NAU SARGASSUM KHANH HOA
THEO T H 6 I GIAN SINH TRlTCtNG
Dang Xuan Cirdnp'. Tran Thi Thanh Van', Bui Minh Ly', Vu Nggc Bgi^
Vign Nghign cira vd U'ng dung Cdng nghe Nha Trang 02A Hung Vuong - Nha Trang, Khdnh Hda
^Trudng Dgi hgc Nha Trang 02 Nguyin Dinh Chilu - Nha Trang, Khanh Hda
Email: [email protected] Tom tat:
Bdi bdo ndy trinh bdy kit qua nghlin ciru sir tich lUy vd phdn bd hdm lugng phlorotannin chdng oxy hda ciia 5 lodi rong ndu (S. polycystum, S. serratum, S.
mcclurei, S. bi'nderi, S. oligocystum) theo thai gian sinh tnrang. 5 lodi rong Sargassum ndy dugc thu mdu theo chu ky sinh tnrang tir thdng 01 din thdng 4/2011 tgi Sdng Ld - Nha Trang, Khdnh Hda vdi sir thir nghiem chdng o.xy hda nhu chdng oxy hda tdng, khu sdt vd khd ndng bdt gdc tu do DPPH. Kit qud th\rc nghiem cho thdy 5 lodi nghiin ciht diu cd hogt tinh chdng oxy hda trin 3 phuang phdp thir nghiem niit trin, trong dd khd nang khir sdt tdt nhdt. Thuc nghiem cung cho thdy tdc dg tich liiy vd phdn bd phlorotannin tuang tu nhu sir tich liiy alginate trong rong vd phii hgp vdi cdc nghiin cim vi thai gian thu hogch rong ndu a Viet Nam va tren the gidi.
ANTIOXIDANT PHLOROTANNIN DISTRIBUTION AND ACCUMULATION IN SOME SARGASSUM BROWN ALGAE SPECIES IN
KHANH HOA ON THE GROWTH TIME Abstract:
This paper show that the research results about antioxidant phlorotannin distribution and accumulation of 5 brown algae species (S. polycystum, S.
serratum, S. mcclurei, S. binderi, S. oligocystum). Those 5 seaweed species were collected from 1/2011 ~ 4/2011 on the growth time at Lo river - Nha Trang city, Khanh Hoa province. The methods were collected to assay antioxidant activity as total antioxidant, reducing power and DPPH radical scavenging. The result showed that 5 species were performed antioxidant activity by 3 aforesaid assay methods, inside the best reducing power. The experiment showed that phlorotannin distribution and accumulation on the growth time are similar alginate distribution and accumulation in seaweed and accord with researchs that research on brown algae harvest time in Vietnam and on the world.
Hgi nghi Khoa hgc vd Cdng nghi bien todn qudc lan thir V
1. MO DAU
Rong ndu thudng tdn tgi mgt lugng ddng kl cac hgp chit polyphenol sinh hgc. Logi duy nhdt trong nhung bdn hgp polyphenolic cd gid tri tir thuc vat bien la "Phlorotannin"
hay la hon hgp polyphenolic trong rong, loai duy nhit dugc bilt tir rong nau [31]. Chiing tdn tgi trong rong ndu dudi dang cdc dan xuit acetate-malonate cua phloroglucinol (1,3,5- trihydroxybenzen) [23], va dugc sinh tong hgp theo con dudng malonate acetate [32], Cdu tnic hda hgc cua phlorotannin trong nhieu loai rong nau da duge nghien cuu bdi [5,6,7,8,9]
va da sd tdn tgi d trgng lugng cao phan tu 40,22,24].
Theo rdt nhilu nghien cira tren the gidi da chi ra polyphenol/ phlorotannin la hgp chat chuyin hda thii cdp gidu boat tinh sinh hgc, trong dd hogt tinh chdng oxy hda da dugc nghien ciiu tren mgt sd loai bdi [12,13,14,26] vd rdt nhieu cdc nha khoa hgc khae. Van dl polyphenol/phlorotannin chdng oxy hda tir rong bien cung da va dang dugc Vien Nghien ciiu va U'ng dung Cdng nghe Nha Trang, Vien Hda hgc cac Hgp chat Thien nhien tien hanh nghien cuu trong nhung nam gan day.
Tuy nhien, ham lugng phlorotannin khac nhau theo'viing dia ly vd thdi gian [17,33].
Trong mgt lodi, phlorotannin cd the tdn tai d cdc te bao sinh san hay te bao md phdn sinh [20,23,28,34]. Ham lugng phlorotannin cd the thay ddi theo miia [23] va bj anh hudng bdi ning d6 cac chat dinh dudng [1,19,35] cudng do anh sang [18,25], chieu cao thiiy trieu [16], vd giai doan phat trien [4], Phlorotannin bi anh hudng bdi alginate cilng dugc chi ra bdi [15]. Khd nang tu bdo ve cd vj tri rdt quan trgng trong he sinh thai va tuang tdc giCta ddng vat an thuc vgt varong [11,28,29],
Vi vay, chiing tdi da tien hanh nghien ciiu su tich liiy va phan bd ciia phlorotaimin theo thdi gian sinh trudng cita mgt so lodi rong ndu thu tai bd bien Khanh Hda, vdi dinh hudng thu nghiem chdng oxy hda nhu chdng oxy hda tdng, khir sdt vd kha nang bat gdc tu do DPPH, ciing su tich liiy cua alginate trong 5 lodi nghien ciiu.
2. DOI TlTQKG VA PHU'CNG PHAP NGHIEN ClTU 2.1. Doi tugng nghien ciiu
5 lodi rong nau 5". polycystum, S. serratum, S. mcclurei, S. binderi, S. oligocystum kha phd bien, phan bd rgng va tap trung chii yeu d duyen hai mien Trung - Viet Nam.
5 loai dugc thu theo thdi gian sinh trudng tir-01 /2011 - 4/201 Id Sdng Ld - Nha Trang - Khdnh Hod. Phan loai va dinh danh bdi chuyen gia phdn loai rong bien TS.Le Nhu Hau.
Sau khi thu mau, rong dugc rira sach bdng nude man, phai khd den ham dm 19% vd bdo quan d nhi?t do thudng.
2.2. Phuang phap nghien cuu
Nghien ciru sutich lily vd phdn bd phlorotannln/polyphenol
Rong dugc thu tirng thdng vd theo cum, vdi sir ngau nhien 3 nhdm trong mdi loai nghien cuu, chieu cao tirng nhom ciia 1 loai cu thi la nhu nhau, trung binh 3 nhdm trong loai la ket qua dugc chgn.
Tieu ban Sinh hoc vd nguon lgi sinh vdt Bien
BS nghien ciiu su tich luy theo thang vi phan b6 trong rong cua phlorotannin, thi lOOg rong tuoi chi6t ethanol 96° trong 24 gia a nhiet do thu&ng, ty le dung moi: nguySn li$u 20:1, roi ti^n hanh dinh lugng phlorotannin, a l ^ a t e va thu hoat tinh ch6ng oxy hoa. Tuy nhiSn, nghien ctiu sir tich luy phlorotannin chSng oxy hoa theo thang thi lay ca cay rong, c6n xac dinh su phSn b6 trong rong thi ISy than da tach la, la tach khoi thto, than hen khfing bo la tach rieng than duoi chua birt la. Sau d6 tiSn hanh cSt nho di ngam chiSt.
Phuong phdp djnh luang
Dinh lirgrng phlorotannin/ polyphenol [31].
Dinh lugng algmate theo Reviers (1989) [27].
Phttangphdp thir hogt tinh
Chong oxy hoa t6ng (TA) dugrc xac dinh theo phuong phap cua Prieto (1999) [21].
Khir Fe (RP) dugc x4c dinh theo phuong phap cua Zhu va cgng su (2002) [36].
Bit g6c tu do DPPH tiSn hanh theo Blois ctmg cong su [2].
Phdn tich vd xir ly s6 lieu
Thi nghiem dugc lap lai (n = 3). Tinh toan do tm c|y cua s6 lieu, phan tich ANOVA va h6i quy bang phin mSm Excell va SPSS 16.
3. KET QUA VA THAO LUAN
Thuc nghiem cho thay S. mcclurei co ham lugrng phlorotannin cao nhat trong 5 loai nghien cuu. S. binderi va S. mcclurei tip trung tich luy phlorotaimin cao vao cu6i tliang 2 V£l dSu t h ^ g 3. S. oligocystum tSp trung tich liiy phlorotannin vao cuoi thdng 3 dSu thdng 4/2011, c6n trong S. seratum va S. polycystum thi phlorotannin lai duac tich luy cao vao cuoi thang 4 diu thang 5/2011. Hinh 1 sau ddy se the hien su tich IGy phlorotannin theo thai gian sinh truang cua 5 loai rong nghien cuu.
phlorotaimin^
/gFA
; Ngaythumai Hinh 1. Bieu dd the hien sif tich lHy phlorotannin theo thai gian sinh trudng 5 lodi rong ndu
Thuc nghiem ciing chi ra ring, trong rong tuai nguyen lieu (Fresh algae - FA) 5.
mcclurei chiia 8,17 mg phlorotannin/g, TA tuang ling vai 6,09mg acid ascorbic/g FA, RP tuang timg 19,lmg FeSOVg FA theo thai gian sinh truang. Tuy nhien, S. mcclurei khong mang hojt tinh chang oxy h6a hay khur Fe cao nhdt, md phdn thuang nay lai thuoc ve loai S oligocystum vai TA tuomg ung vdi 7,32mg acid ascorbic/g FA vao 2/2011, RP ling vcri 51,12mg FeS04/g FA vao thang 3/2011. Dua vdo ket qud cho'thdy, phlorotannm ciia loai
Hgi nghj Khoa hgc vd Cdng ngh? biin todn quoc lan thu V
S. oligocystum cao nhit vao thang 3/2011, hoan toan tuong tu vdi RP cao nhat vao thang nay. Bdng 3.1 sau ddy se thi hien hogt tinh chdng oxy hda cua 5 loai rong nghien ciiu da phdn tich sa luge d tren.
Bdng 1. Hogt tinh chdng oxy hda theo thai gian sinh trudng ciia 5 lodi rong ndu Hoat tinh chong oxy hoa tong (m
Thai gian 15-01-11 15-02-11 15-03-11 15-04-11
5. seratum 2,8679±0,17 2,9264±0,11 3,7917±0,13 4,1677±0,14
5. mcclurei 5,8021±0,08 6,0857±0,09 5,9082±0,24 3,5549±0,26
3, acid ascorbic/g rong tuai nguyen lieu) S. binderi
6,1108±0,32 6,1294±0,26 3,9849±0,31 4,8699±0,27
S. oligocvstum 7,17±0,33 7,3184±0,12 5,3897±0,11 4,4133±0,24
S. polycvsttim 3,5823±0,16 3,9366±0,21 2,2103±0,I8 4,1274±0,19 Hoat tinh khir Fe (mg FeSOVg rong tuai nguyen lieu)
Thai gian 15-01-11 15-02-11 15-03-11 15-04-11
S. seratum 11,1301±0,I4 11,1644±0,11 18,8654±0,08 23,5886±0,16
S. mcclurei 17,762±0,04 19,1019±0,02 20,2231±0,1 3,2781±0,12
iS". binderi I8,1423±0,02 19,7206±0,01 18,9827±0,01 25,8497±0,05
S. olygocystum 12,2619±0,01 12,5502±0,02 5I,1051±0,11 9,9973±0,04
S. polycystum 28,9347±0,02 29,5197±0,01 7,5677±0,04 23,3557±0,09 Ve ho^t tinh bat goc t\r do DPPH, S. mcclurei dat gia trj cao nhdt vdo cu6i thang 2 ddu thang 3/2011, ling vai khoang 93%, nhung S. seratum, S. binderi vd S. oligocystum lai dat 95%, 93% vd 92% vdo cuoi thdng 4 diu thang 5/2011. S. polycystum bit goc tir do gdn nhu hodn todn vdo cuoi thang 1 dau thang 2/2011. Nhu vdy, cho thdy sir phan b3 vd tich luy phlorotaimin dnh huang rdt lan den ho^t tinh chong oxy. hoa trong rong. Tuy nhien doi khi hoat tinh khong dong thiidn vdi su bien doi ciia phlorotannin, diSu do phdn nao cho thay co sv tich luy vd phan b6 phlorotannin co trgng lugng phdn tii khac nhau vdi hoat tinh khac nhau Id khong giong nhau. Thuc nghiem cOng cho thdy, sir tich luy phlorotannin cOn^
tuang ling vai qua trinh tich liiy alginate trong rong, alginate tich liiy trong rong cao nhdt khodng 18% vd chiing dao dgng tir 15,76% - 18,32%. Ve bdn chit phlorotannin t6n tai o dang ngi bao, ngoai bao vd dual dang lien ket vai protein mang te bao vd alginate, nhung tron^ qua triiih tich Ky phlorotannin con chju nhieu tac dgng ciia moi truang dinh duSng va cau tnic he sinh vat thiiy vuc, nen doi khi co su khac bift khSng mang tinh th6ng ke trong tich Wy phlorotannin vd alginate [1,4,11,16,18,19,25,28,29,35].
Ket qua ciing cho thay rang, phlorotannin vd hoat tinh chong oxy hoa cua 5 lodi rong nghien ciiu phdn b6 tap trung a phdn thdn tren han so vai phin thdn dudi, tap trung 6 la nhieu vd phdn than khong co la. Hdm lugng phlorotannin phdn thdn tren cdy rong chiem khoang 60%, phan Id khodng 70%> so yoi toan bg phlorotannin hong rong nghien ciiu. Tir ket qud nay, chiing ta hoan todn co the tien hdnh phan vung phdt triln vimg rong nguyen li$u phuc vii trong thuc phim, dugc phim vd hoach dinh thai gian thu hoach tirng loai, d6ng thai nen cit each g6c chimg 10 - 15cm khi thu hoach [3].
Phdn tich hdm lugng phlorotannin, ho?t tinh chSng oxy hoa ciia phdn thdn tren, than dual, la vd cuong, thiy co s\f tirong quan rit lorn giiia ham lugng phlorotannin, host tinh tirng phdn vai hdm lugng va hojt tinh cua phlorotannin cd cay rong a dp tin cay a < 0,05 ta
Tieu ban Sinh hgc vd ngudn lgi sinh vgt Biin
CO R > 0,98. Hinh 2 sau ddy se the hien c\i the phdn nao nhung bien ludn tren.
mg ^ phlorotannin ? /gFA (,
5 -
a^-Oi-ii j ^ - o j 11 ij-03-i! i^ox II Ngaythumdu Hinh 2. 5v tich liiy vdphdn bd phlorotannin chdng oxy hda theo thai gian sinh truang cua
rong ndu Sargassum seratum
4. KET LUAN
S. binderi va S. mcclurei tdp trung tich IGy phlorotannm cao vao cudi thdng 2 va diu thdng 3, tuang iing vdi 6,77 va 8,17mg phlorotannin/g FA. S. oligocystum tdp trung tich luy phlorotannin vao cudi thdng 3 ddu thang 4/2011, ung vdi 2,9mg phlorotannin/g FA.,^.
seratum va S. polycystum tich luy cao phlorotannin vao cudi thang 4 ddu thdng 5/2011 iing vdi4 va3,5mg/gFA.
Su phdn bd phlorotannin cd boat tmh chdng oxy hda trong 5 loai nghiSn ciiu tdp tnmg khodng 60% d Yz thdn tren, 70% d phdn Id.
Su tich luy va phan bo hoan toan phii hgp vdi cac giai doan sinh trudng cua tiing loai.
Ham lugng phlorotannin tang giam ciing chieu vdi ham lugng alginate trong rong.
LOnCAMOfN
Tgp thi cdc tdc gia chdn thanh cam an chuong trinh nghien cim ca ban dinh hudng iing dyng ciia Bd Khoa hgc va Cdng nghe da cap kinh phi de thuc hi6n dd tdi nay.
TAI LIIU THAM KHAO
1. Arnold TM, Tanner CE, Hatch Wl, Mar Ecol Prog Ser, 123, 177-183, (1995).
2. Blois, M. S. Nanu-e, 26,1199-1200, (1958).
3. Bui Minh Ly va Le Nhu Hau, "Ddnh gid hien trgng vd nghiin ciru gidi phdp bdo v?
ngudn lgi rong ma (Sargassum) tgi Khdnh Hda ", Bdo cao tdng kit dl tdi khoa hgc cdp tinh Khdnh Hda, 12/2010.
4. Denton A, Chapman ARC, Markham J, Mar Ecol Prog Ser, 65, 103-104, (1990).
5. Glombitza K.W. and Sattler E, 43,4277-4280, (1973).
Hdi nghi Khoa hgc vd Cdng nghe bien todn qudc lan thic V
6. Glombitza K.W., Hauperich S. and Keusgen M, Nat. Toxins, 5, 58-63, (1997).
7. Glombitza K.W., Koch M. and Eckhardt G, Phytochemistty, 16, 796-798, (1977a).
8. Glombitza K.W., Rauwald H.W. and Eckhardt G, Phytochemistry 14: 1403-1405, (1975).
9. Glombitza K.W., Rauwald H.W. and Eckhardt G, Planta Med, 32, 33-45, (1977b).
10. Glombitza K.W. and RosenerH.U, Phytochemistay, 13, 1245-1247, (1974).
11. Harvell CD, Q Rev Biol, 65, 323-340, (1990).
12. Kang, H. S., et al., Chemical and Pharmaceutical Bulletin, 51, 1012-1014, (2003).
13. Kang, H. S., et al., Archives of Pharmacal Research, 27, 194-198, (2004).
14. Keejung Kang, Yongju Park, Arch Phaim Res, 26, 4, 286-293, (2003).
15. Leonardo T. S., et al, Brazilian Journal of Oceanography, 55(2), 145-154, (2007).
16. Martinez EA, Hydrobiologia, 326/327, 205-211, (1996).
17. Pavia H, Aberg P, Hydrobiologia, 326/327, 199-203, (1996).
18. Pavia H, Cervin G, Lindgren A, Aberg P, Mar Ecol Prog Ser, 157, 139-146, (1997).
19. Peckol P, Krane JM. Yates JL, Mar Ecol Prog Ser, 138,209-217, (1996).
20. Pfister CA, Ecology, 73, 1586-1596, (1992).
21. Prieto, P., Pineda, M., & Aguilar, M., Analytical Biochemistry, 269, 337-341, (1999).
22. Ragan M.A. and Craigie J.S, L. Can. J. Biochem, 54, 66-73, (1976).
23. Ragan M.A. and Glombitza K.W, Prog. Phycol. Res, 4, 129-241, (1986).
24. Ragan M.A. and Jensen A, J. Exp. Mar. Biol. Ecol. 30: 209-221, (1977).
25. Ragan MA, Jensen A, Le Jol J Exp Mar Biol Ecol, 36, 91-101, (1979).
26. Rastian Zahra, et al, African Jouriial of Biotechnology, 6 (24), 2740-2745, (2007).
27. Reviers, B., J. Appl. Phycol., 1,75-76, (1989).
28. Simms EL. Rausher MD, Am Nat, 130, 570-581, (1987).
29. Steinberg PD, Mar Ecol Prog Ser, 112, 129-133, (1994).
30. Steinberg PD, Science, 223, 405-407, (1984).
31. Swanson, A. K. and Druehl, L. D., Aquatic Botany, 73,241-253, (2002).
32. Targett Nancy M and Arnold Thomas M, Journal of Chemical Ecology, 24,3,1998.
33. Targett NM, Boettcher AA, et al., Oecologia, 103,170-179, (1995).
34. Tugwell S, Branch GM, J Exp Mar Biol Ecol, 129,219-230, (1989).
35. Yates JL. Peckol P, Ecology, 74,1757-1766, (1993).
36. Zhu, Q. T., et al. Journal of Agricultural and Food Chemistry, 50,6929-6934, (2002).