TAP GHJ GONG THIfdNG
XAC DINH DIEU KIEN
CHIET THICH H0P DE THU NHAN BROMOPHENOLS TlT RONG DO LAURENCIA INTERMEDIA YAMADA
» NGUYIN THE HAN - NGUYIN VAN MINH - PHAM THJ HIEN - vO LE QUYEN
TOM TAT:
Bromophenols la nhdm chat cd boat tinh sinh hoc riem nang cd the sijf dung trong ngan ngffa va dieu tri nhieu loai benh. Rong bid'n dtfdc chffng minh la ngu6n nguyen lidu giau bro- mophenols. Nghien ctfu nay danh gia anh htfdng cua dieu kidn chie't de'n hieu qua thu nhan bro- mophenols tij" rong do Laurencia intermedia Yamada. Dieu kien chie't thich hdp dtfdc xac dinh nhtf sau: Dung mdi chid't, ty Id dung mdi chid't/nguydn lieu, nhiet dp chid't, thdi gian chie't lan Itfdl la: 75% methanol, 40/1 (ml/g), 80''C va 90 phut. Ke't qu^ dinh tinh cho thay, dich chid't rong CO cac nhdm chat: phenolics, flavonoids, terpenoids, carotenoids, saponins va tannins.
Ttf khoa: Bromophenols, Laurencia intermedia Yamada, dieu kien chiet, tinh Khdnh Hoa.
l.MdrdSu
Viel Nam nam trong vCing khi hau nhiet ddi vdi bd bien trai dai 3.260 km va didn tich mat ntfdc rong hdn 1.000.000 km^ day la nhtfng dieu kien ttf nhidn thuan ldi cho stf sinh trtfdng va phat trien cua cac loai rong bien. Nhieu cdng trinh nghien cdu trdn the' gidi da chdng minh rang rong bid'n cd chtfa nhieu hdp chat sinh hoc quan trong.
Trong so' do, bromophenols dtfdc chtfng minh la cd nhieu hoat tinh sinh hgc quan trpng va c6 tiem nang su" dung trong thu'c pham chtfc nang, my pham va dtfdc pham. Nhtfng hoat tinh sinh hpc cua bromophenols da dtfdc nghien ctfu la khang te' bao ung thtf, chd'ng oxy hda. khang khuan va didu iri cdc bdnh ve tim mach. Nhieu cha't thudc nhdm bromophenols da dtfdc tim tha'y trong ca ba chi
rong do, rong nau va rong luc thu nhan tai cac vung bien khac nhau trdn the' gidi. Trong dd, cac loai rong do thtfdng cd ham Itfdng bromophenols cao hdn so vdi rong nau va rong luc [1], Cho dd'n nay, cac cdng trinh nghien ctfu ve rong bien d Viet Nam chu ye'u tap trung vao thu nhan mot so' hdp cha't cd ham Itfdng Idn nhtf alginate va agar-agar. Can day, mdt sd' nghien ctfu da btfdc dau thu nhan cac hdp chat cd hoat tinh smh hoc nhtf fucoidan, phlorotannins va polyphenols [2,3].
Tuy nhidn, nhu'ng nghidn ctfu ve nhdm cha't bromophenols tii cdc loai rong bie'n ihu hoach tai vung bien Khanh Hoa ndi ridng va Vidt Nam ndi chung cdn rat ban chd'.
Hieu sua't chie't cac hop chat ttf nhidn phu ihupc vao dieu kien chie't (dung mdi, ly Id dung
206 So21 -Thdna 11/2019
HOA HOC-CONG NCHEIUIfC PHAM
radi/nguyen lieu, nhidt dp chiel va thdi gian chieX—)- Nhffng cdng bd' trtfdc day chi sii dung mdi dieu kidn chid't nha't dinh de thu nhan bromophenols ttf rong bi^n, day cd th^ chtfa phai la dieu kidn chie't thich hdp. Do do, de c6 cd sd cho viec xay dtfng quy trinh thu nhan bromophenols ttf rong bien, can nghien ctfu didu kien chid't thich hdp.
Chi rong Laurencia dtfdc tim tha'y pho bie'n tai cdc viing bien cua Viet Nam, mot so' chat thudc nhom bromophenols c6 hoat tinh sinh hpc da dtfdc tim tha'y trong chi rong nay [4J,
Do do, nghien ctfu nay dtfdc thtfc hien nhdm xdc dinh dieu kien chie't thich hdp de thu nhan dich chie't giau bromophenols ttf rong do Laurencia intermedia Yamada thu hoach lai viing bien Khdnh Hoa,
2. vat lidu va phtfcfng phap nghien citu 2.1. Mdu rong Men vd hoa chdi
Rong do Laurencia intermedia Yamada dtfdc thu nhan tai viing bien Khanh Hoa ttf thang 5 - 7/2017 (Hinh 1). Mlu rong dtfdc dinh danh bdi ThS, Do Anh Duy (Vien Nghien ctfu Hai san, Hai Phong). Sau khi thu nhan, rong dtfdc riia sach bang ntfdc bien va lam khd bang phtfdng phap sa'y lanh d nhiet dp 40''C, de'n dp am dtfdi 13%, Rong khd dtfdc bao gdi chan khdng trong bao hi PA de sii dung cho cac nghien ctfu. Thud'c thu" Foiln-Ciocalleu dtfdc cung ca'p bdi Cdng ty Sigma (Hoa Ky), Cac hoa chat khac su" dung trong nghien cdu deu dat hang phan tich. (Hinh 1)
2.2. Nghien cdu dnh hddng cua dieu kien chie't bromophenols
Anh htfdng cua didu kien chie't ham Itfdng bromophenols thu nhan dtfdc xac dinh bang phtfdng phap cd' dien ddn ye'u td'. Cu the nhtf sau:
- Lua chon he dung moi chie't: Rong nguydn lieu dtfdc chie'l trong dung mdi methanol/ntfdc (0, 25, 50, 75 va 100%, v/v). Cd' dinh cdc thdng so':
nhidt dp chid't, thdi gian chid't va ty le dung mdi/nguyen lieu.
- Lua chon ty li dung mdi/nguyen lieu: Rong nguyen lieu dtfdc chie't trong dung mdi Itfa chpn ttf thi nghidm trdn. Ty le dung mdi/nguyen lidu (ml/g) nghidn ctfu; 10/1, 20/1, 30/1, 40/1 va 50/1. Cd'dinh cac thdng so' cdn lai: nhiet dd chie't va thdi gian chie't.
Li/a chon nhiet do chie't: Rong nguydn lidu dtfdc chie'l trong dung mdi, vdi ly Id dung mdi/nguyen lieu Itfa chon Itf cac thi nghidm tren, Cdc nhiet do chie'l nghien ctfu: 35, 50, 65, 80 va 95''C. Cd'dinh Ihdi gian chie't.
LUa chon thdi gian chiel: Rong nguyen heu dtfdc chie't trong dung mdi, vdi ty le dung mdi/nguyen lieu va nhiet dp chie'l Itfa chon Itt cac thi nghiem trdn. Cac thdi gian chie'l nghien ctfu: 30, 60,90,120 va 150phut.
2.3. Phumg phdp xdc dinh hdm liiffng bromophenols tong
Bromophenols trong rong bie'n dtfdc chie'l theo
Hinh 1: Hinh thai (A) vd hinh cat ngang quan sdt di/dl kinh hien vi (B) mdu rong Laurencia Intermedia Yamada sif dung trong nghien cifu
• v f i H
EL.'^^wv^ lal^^gl
1^'^
nO ' S H H
^B^^^^^^K^*^^ ' 1
TAP CHl CONG THIfiNG
phtfdng phap cua Zhang va cdng stf (2017) [5J.
Ham Itfdng bromophenols long sd' dtfdc xac dinh bang phtfdng phap so mau sii dung thud'c thii Folin - Ciocalteu[6J.
2.4. Dinh tinh mot so' thdnh phdn trong djch chie't
Sau khi chie't d dieu kien chid't thich hdp, mot so' thanh phan trong dich chie'l rong (phenolics, flavonoids, terpenoids, carotenoids, saponins va tannins) dtfdc dinh tinh. Nguydn tac ciia phtfdng phdp dinh linh nay la dtfa trdn phan tfng ddc hieu ciia cha't phan lich va thud'c thii. Mdi thi nghiem dinh dnh lid'n hanh nhtf sau: Lay 0,5 ml dich chie't rong cho vao d'ng nghiem cd chtfa 1,5 ml chat nhan biet. Tie'p theo, quan sdt mau ciia dung dich de' nhan bie't cdc nhdm cha't theo Yadav va cdng stf (2014) [7].
2.5. PhUdng phdp x&ly sd'lidu
Ta't ca cac thi nghidm dtfdc thtfc hidn 3 Ian ddc ISp. Ke't qua ciia thi nghiem dtfdc bie'u didn bang gia trj Irung binh ± dp lech chuan. Do thj dtfdc ve bdng phan mem Microsoft Excel 2010. Gid tri irung binh dtfdc phan tich ANOVA theo phep thii Ducan, gid tri p < 0,05 chi ra stf khde nhau c6 y nghia thd'ng ke sii dung phan mem Statistical Product and Services Solutions (SPSS) 16.0.
3. Ke't qua nghidn cii'u va thao luan 3.1. Anh hddng cua ndng dq dung mdi chiet Anh htfdng cua n6ng dp dung mdi chie't (methanol) de'n ham Itfdng bromophenols long sd' cua mau rong nghien ctfu dtfdc trinh bay d Hinh 2.
Kd't qua cho tha'y, nong dp dung mdi chie't anh htfdng dang ke de'n ham Itfdng cdc p cha't
bromophenols. Khi tdng n6ng dp methanol ttf 0 de'n 75%, ham Itfdng bromophenols tong sd' tang len ttf 48,44 de'n 87,08 mg/kg khd'i Itfdng khd. Tid'p luc tang nong do methanol len 100%, ham Itfdng bromophenols t6ng sd' giam xud'ng cdn 74,39 mg/kg khd'i Itfdng khd.
Trong nhtfng nghidn ctfu trtfdc day ve thu nhdn bromophenols ttf rong bien, cac tac gia thtfdng sijf dung mpi loai dung mdi nha't dinh. Acetone, eihanol va methanol hodc hdn hdp cua cac dung mdi nay trong ntfdc (thtfdng la 80%), thtfdng dtfdc suf dung de chie't cac hdp chat bromophenols [4,8,9]. Trong nghien ctfu nay, cac nong dp dung mdi methanol khac nhau dtfdc nghien ctfu. Ke't qua nghidn ctfu btfdc dau nay cd the tha'y 75%
methnanol cho hieu qua chie't cac cha^t bromophenols ttf rong la td't nhat trong khoang nong dp dung mdi nghidn ctfu. (Hinh 2)
3.2. Anh hudng cua ty 1$ dung mdi chiet/
nguyen lieu
Anh htfdng ciia li le dung moi chid't/nguydn lieu de'n ham Itfdng bromophenols dtfdc the hien 6 Hinh 3. Ke't qua nghien ctfu cho thay dung mdi chie'i/nguyen lieu 40/1 (ml/g) la thich hdp cho qua trinh chie't de thu dtfdc ham Itfdng cao nhd't bromopphenols. Ti le nay cho ham Itfdng bromophenols (104,96 mg/kg khd'i Itfdng khd) cao hdn dang k^ so vdi chie't ct ti Id 10/1 va 20/1 (ml/g).
Chid't d ti le dung moi chid't/nguydn heu 50/1 khdng lam lang ddng ke' ham Itfdng bromophenols (p >
0,05). Ttf nhtfng ke't qua dai dtfdc, ti Id dung mdi chiel/nguyen lieu la 40/1 (ml/g) dtfdc Itfa chpn cho nhtfng thi nghidm lie'p theo. (Hinh 3)
Hinh 2: Anh hifdng cua ndng dp dung mdi chie't methanol den hdm lifdng bromophenols trong dich chief rong Laurencia Intermedia Yamada. Cdc chif cdi khac nhau chi si/khde nhau CO y nghia thd'ng ke (p > 0,05)
100 n .,0 9 0 •
S s o •
"1 7 0 •
1 1 60
-a-<ts 50 • O -S 3 " 40
O BIj
i f 3°
g - S 20 •• = 1 " "
•S 0 •
a
i A ^ •
<> ^ 1 1
^ • • •
^^1 ^^1 ^^1 ^^1
1 I 1 1
H H • H
1 1 1 1
• , • , • , •
0 25 SO 75 Nong dp methanol (%)b
^ •
• 1
• I
M .
100208 So 21 -Thang 11/2019
HOA HOC-CONG NGHE THifC PHAM
Hinh 3: Anh hifdng ciia ty le dung mdi/nguyen lieu den hdm lUdng bromophenols trong djch chief rong Laurencia Intermedia Yamada. Cac chif cai khde nhau chi sif khde nhau CO y nghla thdng ke (p < 0,05)
^^
30/1 40/1 Ty le dung mdi/nguyen lieu (ml/g)
3.3. Anh hifdng cua nhiet do chie't
Anh htfdng ciia nhiel do chie'l de'n ham Itfdng bromophenols td'ng sd ciia loai rong nghien ctfu dtfdc the hien d Hinh 4. Ke'l qua cho Ihay, nhidt do chie't anh htfdng ddng ke de'n ham Itfdng cac hdp cha't bromophenols td'ng so'. Anh htfdng ciia nhidt do dd'n h^m Itfdng bromophenols nhtf sau:
Khi nhidt dp chid't tang ttf 35 len SO^C, ham Itfdng bromophenols tang manh ttf 54,78 de'n 102,65 mg/kg khd'i Itfdng khd. Tid'p tuc tang nhiel dp len 95*'C ham Itfdng bromophenol giam nhe xud'ng con 97,46 mg/kg nguyen Hdu khd va khdng cd stf khac nhau cd y nghia ve mat thd'ng kd vdi ham Itfdng bromophenols long so' khi chid't d nhiet do
SO'^C (p > 0,05). Cho de'n nay, nhffng nghidn ctfu ve dieu kidn chid't dp thich hdp d^ thu nhan bromophenols ttf rong bi^n con rd't ban chd'. Mdt sd' nghidn ctfu anh htfdng ciia nhiel dd chie'l dd'n ham Itfdng polyphenol long so' trong dich chie'l iff thtfc v3t cho ket qua ttfdng itf vdi nghien ctfu nay, Vi du, Bucic-Kojic va cdng stf (2011) [10] khi nghien ctfu anh htfdng ciia nhiet dp chie'l dd'n ham Itfdng polyphenol td'ng so ciia trdi va (Ficus Carica L.), cho tha'y ham Itfdng tang len ttf 2,50 de'n 3,70 mg GAE/g cha'l khd khi tang nhidt dd chie'l Itf 25 len SO^C. Nhtf vay, SO^'C dtfdc Itfa chon la nhiet dp thich hdp nha't trong khoang nhietdp nghien ctfu. (Hinh 4)
hTinh 4: Anh hudng cua nhiet do chief den hdm lUdng bromophenols trong djch chief rong Laurencia intermedia Yamada. Cdc chif cai khac nhau chi sif khde nhau cd y nghia thdng ke (p < 0,05)
120 -
•<o OL
Jsl""-
-s S 2 00 80 -
J 1
9"-! 60 - d - o S 2 » » 40 -
&>
,2 1 2 0 n .g K 0 .
d
A •
H
35
a
± I
^ ^ 1 ^H^ 1 1 H • I H I
I I I
I I I
, M > ^ , *
50 65 80 Miiet do (°C)
a
^
1 1
• I
1 , 1 H
95TAP CHi CONG THIflfNG
3.4. Anh hddng cua thdi gian chiet Ben canh nhiet dp va dung mdi chie't, Ihdi gian cung la ye'u Id' quan trpng anh htfdng dd'n hieu qua chie't cac hdp chat sinh hpc ttf nguyen lieu itf nhien. Anh htfdng cua Ihdi gian chie't de'n ham Itfdng bromophenols td'ng sd' ciia rong dtfdc the' hidn d Hinh 5. Ke't qua cho tha'y, thdi gian chie'l anh htfdng ddng ke dd'n ham Itfdng cdc chat bromophenols.
Khi thdi gian chid't tang ttf 30 len 90 phut thi ham Itfdng bromophenols long so' lang manh. Tuy nhien, khi tie'p tuc tang thdi gian chie't len 90 va 150 phiil thi ham Itfdng bromophenols khdng de'p tuc tang, khdng cd stf khac nhau cd y nghia ve mat thong ke (p > 0,05) so vdi Ihdi gian chid't 120 phut.
Nhtf vay, theo kd't qua nghien ctfu thdi gian chie't thich hdp dd' ihu nhan bromophenols ttf rong dtfdc xac dinh la 90 phiil. Trong nhtfng nghien ctfu trtfdc day, vi chtfa dtfdc danh gid mdt each day dii, nen thdi gian chie't
bromophenols thtfdng keo dai [8,9]. Dieu nay, CO the' gay lang phi thdi gian va dung mdi chid't.
(Hinh 5)
3.5. Nh^n biet mdt sd' nhom chat cd trong dich chiet rong
Ke'l qua dinh tinh mot sd' thanh phan phd' bien CO trong dich chid't Laurencia intermedia Yamada cho tha'y, ta't ca cac nhdm chat nghien ctfu (alkaloids, phenolics, flavnooids, carotenoids, terpenoids, saponins va tannins) ddu cd trong dich chie'l rong.
Kd't qua nghien cffu nay phu hdp vdi mdt sd' loai rong thudc chi Laurencia thu mdu tai cdc viing bien khde nhau trdn the'gidi [11,12]. Thanh phan cac nhdm chat c6 trong rong bidn phu thudc vao dieu kien tdch chie'l va tac ddng bdi cdc yd'u to' mdi trtfdng sd'ng cua rong [II]. Stf xua't hien cua cdc nhom chat cd trong djch chie'l rong Laurencia intermedia Yamada da giai Ihich cho nhffng hoat tinh sinh hoc ciia loai rong nay. Tuy nhien, nhffng nghien ctfu lid'p theo can dinh Itfdng nhffng nhom chat nay.
4. Ke't luan
Nghidn ctfu hidn tai da xdc dinh dtfdc dieu kien chie'l thich hdp de thu nhdn bromophenols ttf rong do Laurencia intermedia Yamada Ihu hoach tai vung bie'n Khanh Hoa. Nhffng nghien ctfu tid'p theo can thtfc hien de' td'i tfu hoa dieu kien thu nhan va xay dtfng quy Irinh dnh sach bromophenols ttf loai rong nay •
Hinh 5: Anh hifdng cua fhdi gian chief de'n hdm lifdng bromophenols trong djch chief rong Laurencia infermedia Yamada. Cdc chit cdi khde nhau chi sU khac nhau c6 y nghia fhd'ng k§ (p > 0,05)
-s ^^° 1
00
• 1 ^ 100 •
•S ^ a M 8 0 •
1-
1 . | 60 -"toj^ 40 -
&>
• 3 S 2 0 -
.1
a; 0 _
a a
T ^ ^ ^
I" • • m m
^ • •
^ • 1 1 I 1 1 1
1 1 1 1
1 1 1 1
1 • . • , • . • ,
30 60 90 120 Thai gian chiet (phut)a
ii •
• 1 •
1 •
1
150Ldi cdm dn:
Nghien cdu nay dddc tdi trd bdi de tai kitoa hoc cdng ngh§ linh Khdnh Hoa, md sd': DT-2017-20902-DL.
210 So21 -Thang 11/2019
HOA HOC-CONG NGHE IHIfC PHAM
TAI LIEU THAM KHAO:
7. Whitfield. F. B.. Helidoniotis. F.. Shaw. K. J.. & Svoronos. D. (1999). Distribution of bromophenols in speeles of marine algae from eastern Australia. Journal of Agricultural and Food Chemistry. 47(6),
2367-2373.
2. Thinh. P. D., Menshova. R. V., Ermakova, S. P., Anastyuk, S. D., Ly, B. M.. & Zvyaginlseva. T. N.
(2013). Structural characteristics and anticancer activity of fucoidan from the brown alga Sargassum mcclurel. Marine Drugs, 11(5), 1456-1476.
3. Cuong. H. D., Thuy, T. T. T., Huong, T. T.. Ly. B. M.. & Van, T. T. T. (2015). Structure and hypollpldaemic activity of fucoidan extracted from brown seaweed Sargassum henslowianum. Natural Product Research, 29(5), 411-415.
4. Mikaml. D., Kurlhara, H., Kim, S. M.. & Takahashl, K. (2013). Red algal bromophenols as glucose 6- phosphale dehydrogenase Inhibitors. Marine Drugs. 11(10). 4050-4057.
5. Zhang, S.. Li, Y., You. ]., Wang, H.. Zheng, ¥., & Sua, Y. (2012). Improved method for the extraction and determination of bromophenols in seafoods by high-performance liquid chromatography with fluorescence detection. Journal of Agrictdturaland Food Chemistry, 60(44), 10985-10990.
6. Fu. L, Xu, B. T, Gan, R. Y., Zhang, Y.. Xu, X. R.. Xia. E. Q., & U H. B. (2011). Total phenolic contents and antioxidant capactties of herbal and tea Infusions. International Journal of Molecular Sciences, 12(4), 2112-2124.
7. Yadav, M., Chatterji, S., Gupta. S. K.. & Watal. G. (2014). Preliminary phytochemical screening of six medicinal plants used in traditional medicine. International Journal of Pharmacy and Pharmaceutical Sciences, 6(5), 539-542.
8. Xu, X. L, Fan, X.. Song. F. H., Zhao. J. L, Han. L. J., Yang. Y. C, & Shi, J. G. (2004). Bromophenols from the brown alga Leathesla nana. Journal of Asian Natural Products Research, 6(3), 217-221.
9. Kim. K. Y, Nguyen. T. H.. Kurihara, H.. & Kim, S. M. (2010). a - glucosldase Inhibitory activity of bromophenol purified from the red alga Polyopes lanclfolia. Journal of Food Science, 75(5), H145-H150.
10. Buclc-Kojlc, A, Planlnic. M., Tomas. S, Jokic, S., Mujlc, I., Billc, M., 4 Velic. D. (2011). Effect of extraction conditions on the extractabillty of phenolic compounds from lyophllisedfigfruljs (Ficus carica L). Polish Journal of FoodandNutrition Sciences. 61(3). 195-199.
11. Jeyaseetan. E. C, Kothai. S., Kavitha, R.. Tharmila. S. & Thavaranpl. A. C. (2012). Antibacterial activity of some selected algae present In the costal lines of Jaffna peninsula. International Journal of Pharmaceutical & Biological Archives. 3(2), 352-356.
12. Omar, H.. Al-Judaibiand. A., & El-Cendy. A. (2018). Antimicrobial. Antioxidant. Anticancer Activity and Phytochemical Analysis of the Red Alga. Laurencia papillosa. International Journal of Pharmacology. 14(4). 572-583.
Ngay nhan bai: 12/10/2019
Ngay phan bi$n danh gia va su'a chffa: 22/10/2019 Ngay cha'p nh$n dang ball 2/I1/2D19
TAP CHl CONG THIflfNG
Thong tin ve tdc gla:
1, TS, NGUYfeN THE HAN 2, PGS,TS, NGUYEN VAN MINH 3, ThS, PHAM THI HIEN 4,ThS,VULEQUYEN
Khoa C6ng nghe Thffc phS'm, Trfftog Dai hpc Nha Trang
DETERMINATION OF SUITABLE EXTRACTION CONDITIONS FOR EXTRACTING
BROMOPHENOLS FROM RED SEAWEED LAURENCIA INTERMEDIA YAMADA
• PHD NGUYEN THE HAN
• Assoc. ProF. PHD NGUYEN VAN MINH
• MSC, PHAM THI HIEN
• MSC.VULE QUYEN
Faculty of Food Technology - Nho Trang University
ABSTRACT:
Bromophenols are potential bioactive compounds and can be used for prevention and treatment of various diseases. Seaweeds have been demonstrated as rich sources of bromophenols. This study investigated effects of extraction conditions on bromophenols content of red seaweed Laurencia Intermedia Yamada. The suitable solvent extraction, hquid to soHd ratio, extraction temperature and extraction time were found as follows: 75% methanol, 40/1 (ml/g), 80°C and 90 min, respectively. Phytochemical analysis of the seaweed extract showed that phenolics, flavonoids, terpenoids, carotenoids, saponins and tannins are presented.
Keywords: Bromophenols, iMurencla Intermedia Yamada, extraction conditions, Khanh Hoa provin.
212 So 21 -Thdng 11/2019