LI CM N

Tri qua thi gian hc tp ti trng v 3 thng lm n tt nghip ti Vin nghin cu v ng dng Nha Trang, ti hon thnh bi n: Nghin cu trch ly phlorotannin t rong Nu Sargassum Serratum v ng dng to sn phm ung giu phlorotannin. t c kt qu , trc ht ti xin by t lng bit n su sc n PGS.TS Bi Minh L ngi tn tnh gip ti trong sut thi gian ti tin hnh thc hin ti. Ti chn thnh cm n qu Thy, C Khoa Cng ngh Thc Phm trng i hc Nha Trang tn tnh dy d, ch bo ti trong sut thi gian hc ti trng. Xin gi li cm n n TS. Trn Th Thanh Vn, Th.S V Mai Nh Hiu, Ks. Trn Nguyn H Vy cng cc anh ch cng tc ti phng th nghim ha phn tch v trin khai cng ngh ca Vin tn tnh hng dn, gip ti trong sut qu trnh thc hin n ny. Ti xin gi li cm n n gia nh, bn b l ch da vng chc, ht lng ng vin gip ti trong sut qu trnh hc tp cng nh thc hin ti. Mt ln na ti xin chn thnh cm n!

Nha Trang, thng 6 nm 2012 Sinh vin thc hin Hong Th Thy An

MC LC MC LC ...............................................................................................................I LI M U .........................................................................................................1 CHNG 1: TNG QUAN .................................................................................3 1.1. TNG QUAN V RONG BIN [1]. ............................................................3

1.1.1 c im, s phn b rong bin. ...............................................................3 1.1.2 Ngun li rong bin Bo v v pht trin ngun li rong bin Vit Nam.........4 1.1.3 Phn loi rong bin.....................................................................................7 1.1.4 Thnh phn sinh ha ca rong bin ............................................................8 1.1.5 Gi tr dinh dng v ng dng ca rong bin. ...........................................8 1.1.6 c im sinh trng, pht trin v thi k thu hoch rong bin hp l cho cng ngh ch bin. ........................................................................................... 10 1.1.7 Tnh hnh s dng, ch bin rong bin Vit Nam.................................. 12 1.2 1.3 Qu trnh vn chuyn v cc bin php bo qun rong kh. ........................ 12 Gii thiu v rong Nu v rong M (Sargassum)........................................ 13

1.3.1 c im. ................................................................................................ 13 1.3.2 S phn b [1]. ........................................................................................ 16 1.3.3 Thnh phn ha hc ca rong Nu. ......................................................... 18 1.4 Mt s quy trnh cng ngh sn xut cc cht t rong Nu. ........................ 24

1.4.1 Cng ngh ch bin Mannitol t rong Nu [1]. ........................................ 24

1.4.2

Chit rt Iod t rong Nu [1] ....................................................................... 25

1.5

Tng quan v hp cht Phlorotannin (polyphenol). ..................................... 26

1.5.1 c im. ................................................................................................ 26 1.5.2 C ch oxi ha ca polyphenol................................................................ 27 1.5.3 Hot tnh sinh hc ca phlorotannin. ....................................................... 28 1.5.4 ng dng. ............................................................................................... 29 1.6 Gii thiu v qu trnh trch ly. ................................................................... 30

1.6.1 Bn cht. ................................................................................................. 30 1.6.2 Phm vi s dng ca qu trnh. ............................................................... 30 1.6.3 Cc yu t nh hng n qu trnh chit. ............................................... 31

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1.6.4 Tng quan v dung mi chit. ................................................................. 32 1.6.5 Tm hiu dung mi chit trong ti. ...................................................... 34 CHNG 2: I TNG V PHNG PHP NGHIN CU .................. 37 2.1 2.2 2.3 2.4 i tng nghin cu. ................................................................................ 37 Ha cht v dng c th nghim. ................................................................. 37 Phng php nghin cu ............................................................................ 37 Quy trnh trch ly v thu nhn phlorotannin................................................. 41 S quy trnh....................................................................................... 41

2.4.1

2.4.2 Thuyt minh quy trnh............................................................................. 42 2.5 Cc yu t nh hng n qu trnh trch ly Phlorotannin. .......................... 43

2.5.1 Tin hnh kho st iu kin mi trng (pH) trch ly............................. 43 2.5.2 Tin hnh kho st t l dung mi trch ly. .............................................. 44 2.5.3 Tin hnh kho st nhit trch ly. ........................................................ 45 2.5.4 Tin hnh kho st thi gian trch ly. ....................................................... 46 2.5.5 Tin hnh xc nh hiu sut trch ly ....................................................... 47 2.5.6 Th nghim xc nh nhit c c. ...................................................... 48 2.5.7 Th nghim xc nh thi gian ly tm ...................................................... 49 2.6 ng dng cao chit phlorotannin vo qu trnh to sn phm: nc ung

rong bin ng chai. .............................................................................................. 50 CHNG 3: KT QU NGHIN CU V THO LUN ............................ 52 3.1 3.2 3.3 Kt qu xc nh mt s thnh phn ca rong nguyn liu. ......................... 52 Kt qu kh mi ca rong bin. .................................................................. 53 Kt qu xc nh thng s thch hp ca qu trnh chit phlorotannin. ........ 53

3.3.1 Kt qu xc nh nh hng ca mi trng (pH) n kh nng chit. .... 54 3.3.2 Kt qu xc nh nh hng ca t l NL/DM (khi lng/th tch) n kh nng chit. ................................................................................................... 55 3.3.3 Kt qu xc nh nh hng ca nhit n kh nng chit. ................. 58 3.3.4 Kt qu xc nh nh hng ca thi gian n kh nng chit. ................ 59 3.3.5 Xc nh hiu sut trch ly. ...................................................................... 61 3.3.6 Kt qu xc nh thng s thch hp ca qu trnh c c........................ 63

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3.3.7 Kt qu xc nh thng s thch hp ca qu trnh ly tm........................ 66 3.3.8 xut quy trnh chit phlorotannin t rong Sargassum Serratum.......... 67 3.3.9 ng dng cao Phlorotannin vo qu trnh to sn phm. ......................... 69 KT LUN V XUT KIN .................................................................... 74 TI LIU THAM KHO ..................................................................................... 77 PH LC..................................................................................................................

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DANH MC BNG Bng 1.1: Ngun li, sn lng thu hoch v tim nng sn xut rong bin [1]. ......5 Bng 1.2: Din tch cc vng c th qui hoch nui trng rong bin mt s tnh duyn hi Vit Nam [1]. ..........................................................................................6 Bng 1.3: Cc ging loi rong Nu tm thy v phn b. ....................................... 16 Bng 1.4: Tr lng rong M theo vng bin cc tnh. .......................................... 17 Bng 1.5: Hm lng Mannitol 2 loi rong S.mcclurei v S.kjellmanianum ti Hn Chng Nha Trang 1979 (% so vi trng lng kh tuyt i)................................ 18 Bng 1.6: Hm lng axit Alginic trong cc loi rong Nu ................................... 20 Bng 1.7: Hm lng axit amin mt s loi rong Nu vung bin Jeddah, Saudi Arabia (mg %)....................................................................................................... 22 Bng 1.8: Hm lng Iod trong cc loi rong Nu (% trng lng kh) Nha Trang Khnh Ha. ........................................................................................................... 23 Bng 2.1: Cc mc cht lng .............................................................................. 39 Bng 2.2: H s trng lng ca sn phm............................................................ 40 Bng 3.1 Thnh phn ha hc chnh ca rong nguyn liu .................................... 52 Bng 3.2 nh hng ca nng dung dch H2O2 n kh nng kh mi............ 53 Bng 3.13: nh hng ca nhit n hm lng Phlorotannin v Carbohydrate trong qu trnh c c. ........................................................................................... 64 Bng 3.14: Hm lng cc thnh phn (%) sau khi c c..................................... 65 Bng 3.15 Kt qu nh gi cm quan dch chit. ................................................. 66 Bng 3.16: Ch tiu ha l v nht ca mt s sn phm nc ngt.................. 69 Bng 3.17: Kt qu nh gi cm quan dch chit theo t l dch cao b sung. ...... 69 Bng 3.18: Bng nh gi cm quan dch chit theo t l ng phi ch ............. 70 Bng 3.19: Kt qu ch tiu vi sinh theo thi gian thanh trng............................... 71 Bng 3.20: Bng m t sn phm nc rong bin ng chai c b sung cao phlorotannin. ......................................................................................................... 71 Bng 3.21: Bng phn tch cc thnh phn ca sn phm sau khi b sung cao phlorotannin. ......................................................................................................... 72 Bng 3.22: Kt qu kim tra ch tiu vi sinh vt trong sn phm............................ 72

DANH MC HNH Hnh 1.1: S hnh chit Mannitol bng phng php ha tan trong Alcol ..........25 Hnh 1.2: S quy trnh tng qut chit rt Iod t rong Nu................................... 26 Hnh 1.3: Phloroglucinol (i) v phlorotannin [tetrafucol A (ii), fucodiphloroethol B (iii), fucodiphlorethol A (iv), tetrafuhalol A (v), tetraisofuhalol (vi),

phlorofucofuroeckol (vii)] v hot tnh ca chng. ................................................27 Hnh 2.1: Rong M (Sargassum serratum) .............................................................37 Hnh 2.2: S quy trnh thu nhn phlorotannin t rong Sargassum Serratum. ......41 Hnh 2.3: S b tr th nghim xc nh nh hng ca pH ................................44 Hnh 2.4: s b tr th nghim xc nh nh hng ca t l dung mi n kh nng trch ly. ..........................................................................................................45 Hnh 2.5: S b tr th nghim xc nh nh hng ca nhit ........................46 Hnh 2.6: S b tr th nghim xc nh nh hng ............................................47 Hnh 2.8: S b tr th nghim xc nh nhit c c. ....................................48 Hnh 2.9: S b tr th nghim xc nh thi gian ly tm ....................................49 Hnh 2.10: S quy trnh to sn phm nc ung rong bin ng chai...............50 Hnh 3.1a: Biu biu din nh hng ca pH trch ly n hm lng phlorotannin trong qu trnh chit ..........................................................................44 Hnh 3.1b: Biu biu din nh hng ca pH trch ly n hm lng carbohydrate trong qu trnh chit. .........................................................................55 Hnh 3.2a: Biu biu din nh hng ca t l NL/DM n hm lng phlorotannin trong qu trnh chit. ..........................................................................56 Hnh 3.2b: Biu biu din nh hng ca t l NL/DM n hm lng carbohydrate trong qu trnh chit. .........................................................................56 Hnh 3.3a: Biu biu din nh hng ca nhit chit n hm lng phlorotannin trong qu trnh chit. ..........................................................................58 Hnh 3.3b: Biu biu din nh hng ca nhit chit n hm lng carbohydrate trong qu trnh chit. .........................................................................58

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Hnh 3.4a: Biu biu din nh hng ca thi gian chit n hm lng phlorotannin trong qu trnh chit. ..........................................................................60 Hnh 3.4b: Biu biu din nh hng ca thi gian chit n hm lng carbohydrate trong qu trnh chit. .........................................................................60 Hnh 3.5a: th biu din hm lng phlorotannin trong cc ln trch ly ............61 Hnh 3.5b: th biu din hm lng carbohydrate trong cc ln trch ly. ..........62 Hnh 3.6a: th biu din hiu sut trch ly phlorotannin trong cc ln trch ly ....62 Hnh 3.6b: th biu din hiu sut trch ly carbohydrate trong cc ln trch ly...63 Hnh 3.7: S quy trnh chit rt phlorotannin t rong Sargassum Serratum Khnh Ha. ............................................................................................................67

LI M U

X hi ngy cng pht trin, i sng con ngi ngy cng c nng cao. Con ngi khng ch quan tm n nhng vn n, mc, n gin nh trc y m cn c nhng yu cu cao hn. Khi i sng c nng cao, con ngi ngy cng quan tm hn n sc khe ca mnh. Do , nhng thc phm, vt dng c th gy hi cho sc khe dn b loi b v c thay th bng cc sn phm c sn xut t cc thnh phn chit xut t thin nhin. Nhu cu ca con ngi l mt trong nhng yu t quan trng thc y x hi pht trin v cng l ng lc thc y cc nh khoa hc, nh sn xut tm ti, sng to ra nhiu sn phm mi. Vi s pht trin ca khoa hc, con ngi bit cch chit xut ra nhiu hp cht c ngun gc t nhin c li cho sc khe con ngi v ng dng chng vo trong i sng thng qua nhiu ngnh cng nghip nh: cng nghip thc phm, cng nghip xy dng, dt mayTrong , vic ng dng cc thnh tu khoa hc vo ngnh cng nghip thc phm ng vai tr quan trng v n nh hng trc tip n sc khe con ngi. Hin nay, c rt nhiu hp cht c chit xut t thin nhin c ng dng rng ri vo i sng. Tht vy, sn phm ung khng cn n thun l gii kht m cn phi tt cho sc khe. V th vic nghin cu sn xut cc chit xut t thin nhin b sung vo ung lun c quan tm, v nhng cht c trch ly chnh l hp cht polyphenol (phlorotannin). Thnh phn ny c s dng trong thc phm nh mt loi thc phm chc nng nhm mc ch phng nga bnh do c tnh cht khng oxi ha mnh. Vit Nam c h ng vt, thc vt v cng phong ph, c nhiu gen qu him c trng cho kh hu nhit i nng m. Mt trong nhng iu kin to nn s phong ph v giu c y chnh l vng bin nhit i rng vi b di hn 3200 km bao bc ht pha ng v nam t nc. Mt trong nhng ngun ti nguyn phong ph v c gi tr m vng bin ban tng cho chng ta l rong bin. Rong bin l loi thc vt bin qu gi c dng lm nguyn liu ch bin thnh cc sn phm c gi tr trong cng nghip v thc phm. T lu, rong bin c coi l i tng nghin cu ca nhiu nc trn th gii. nc ta tr lng rong bin

rt ln, l ngun ti nguyn bin v cng phong ph, rong bin chim v tr quan trng trong lnh vc kinh t bin Vit Nam. Ngnh rong bin c nhiu loi, mt trong nhng loi c nhiu tnh nng u vit c nhiu nh nghin cu nc ta quan tm ti l ngnh rong Nu m in hnh l rong M. Gn y, ngnh nui trng v ch bin rong bin ni ln nh mt ngnh cng nghip mi mang li mt s thnh tu nht nh. Nhiu cng trnh nghin cu v gi tr dinh dng cng nh dc hc t rong bin c cng b v ng dng rng ri trn ton cu. Ngi ta pht hin ra nhiu thnh phn qu c trong rong bin nh: Iod, Alginate, Fuccoidin, hp cht chng oxi ha (phlorotannin), cc axit bo,... rt c gi tr trong y hc, thc phm, dc phmgip cho mi quc gia gii quyt c vn nhp khu dc liu. Ngy nay, nhiu cng dng khc ca rong bin cn ang c cc nh nghin cu tip tc khm ph. Xut pht t nhu cu ca x hi hin i, ng thi tn dng c ngun nguyn liu di do, cc nh khoa hc ca nc ta v ang tng cng nghin cu, chuyn ngun rong bin v ph thi t rong bin thnh cc sn phm c gi tr, bng cch sn xut ra cc loi thc phm khc nhau, tch chit ra cc thnh phn khc nhau c hot tnh sinh hc hoc hn hp cc thnh phn c hot tnh sinh hc gia tng gi tr rong bin Vit Nam. gp phn vo xu th , ti thc hin ti Nghin cu trch ly phlorotannin t rong Nu Sargassum Seratum v ng dng to sn phm ung giu phlorotannin. ti gm cc ni dung: 1. Tm hiu mt s thnh phn ha hc ch yu ca nguyn liu rong Nu. 2. Kho st qu trnh trch ly phlorotannin. 3. Th nghim to sn phm ung giu phlorotannin. Bc u tip cn vi nghin cu khoa hc, trong iu kin kin thc cn hn ch, iu kin c s vt cht, kinh ph nghin cu cn thiu thn, ngoi vic n lc ca bn thn, s gip ca gio vin hng dn cng cc anh ch ph trch phng th nghim ca Vin Nghin cu v ng dng cng ngh Nha Trang, ti hon thnh ti c giao. Tuy nhin ti khng trnh khi nhng thiu st, ti rt mong c s gp chn thnh ca cc thy c, cc bn c gi ti c hon thin hn. Ti xin chn thnh cm n!

CHNG 1: TNG QUAN

1.1. TNG QUAN V RONG BIN [1]. 1.1.1 c im, s phn b rong bin. Rong bin hay to bin c tn khoa hc l marine algae, marine plant hay seaweed. Rong bin l thc vt thy sinh c i sng gn lin vi nc. Chng c th l n bo, a bo sng thnh qun th. Chng c kch thc hin vi hoc c khi di hng chc mt. Hnh dng ca chng c th l hnh cu, hnh si, hnh phin l hay hnh th rt c bit. Sn lng hng nm cc a dng cung cp cho tri t hng 200 t tn rong. Nhiu nh khoa hc cho rng trn 90% carbon tng hp hng nm nh quang hp trong mi trng lng, trong c 20% do rong bin tng hp nn. Rong bin thng phn b cc vng nc mn, nc l, ca sng, vng trin su, Rong v rong Nu l hai i tng c nghin cu vi sn lng ln v c ng dng nhiu trong cc ngnh cng nghip v i sng. i vi rong Lc th loi to Chlorella c xp vo loi to k diu, c tc sinh khi cc nhanh, ang c nghin cu phc v cho con ngi. Rong bin sng bin, hp th mt lng thc n phong ph chy tri dt t lc a ra, rong c nhiu tnh cht khng ging thc vt trn cn. Mt s yu t sinh thi i vi ng vt sng trn cn l rt quan trng, song i vi rong bin li khng quan trng nh m ca khng kh, lng ma. Nhng cc yu t sinh thi bin c nh hng n i sng rong bin nh: a bn sinh trng, nhit , nh sang, mui, PH, mui dinh dng, kh ha tan, mc triu, song, gi, hi lu. Nhiu hp cht hu c trong rong bin c tc dng iu ha, kch thch sinh trng i vi cy trng nh auxin, giberelin, cytokinin, mannitol v cc oligosacaride khc. Ngy nay rong bin cn c sn xut thnh phn bn hu c (phn bn l v phn bn gc).

1.1.2 Ngun li rong bin Bo v v pht trin ngun li rong bin Vit Nam. a. Ngun li rong bin th gii. Ngun li rong bin trn th gii rt ln, song sn lng rong c khai thc v s dng hng nm khng u (theo ti liu ca FAO v sn lng rong bin hng nm trn th gii). Chu l khu vc cung cp rong . Trong , Philippines k t nm 1970 sau khi p dng thnh cng phng php pht trin rong Eucheuma bng bao t chuyn ln hng u th gii v rong bin nguyn liu, 85% lng nguyn liu sn xut Carrageenan v Furcellaran hng nm do Philippins cung cp. Nam Triu Tin l nc cung cp nguyn liu sn xut Agar vi khi lng ln nht trn th gii, chim 52%. Ngun li rong Nu ch yu tp trung cc nc Chu u v Bc M. Canada tp trung hn 75% khi lng rong nguyn liu sn xut Alginate , trong khi khi lng rong Nu Chu ch khong 5%. Theo FAO c tnh mi nm trn th gii rong Nu c khai thc dc b i Ty Dng k c bin en v a Trung Hi [1]. Trn th gii Alginate c sn xut t rong Nu c sn lng ln hn Agar, Carrageenan, Furcellanan c sn xut t rong . V sn lng rong Nu th khu vc Bc M c sn lng ln nht, tip n l Chu u, M La Tinh v Chu . i vi rong th sn lng ch yu tp trung ln ti Chu , n Chu M La Tinh, ri n Chu u. Vic chn loi rong no lm nguyn liu chnh sn xut cc loi keo rong ph thuc vo nhiu yu t khc nhau. Mt trong cc yu t quan trng nht l tnh cht n nh v ngun nguyn liu, hay ni mt cch khc l ph thuc vo kh nng pht trin ca loi rong trong iu kin t nhin ca mi nc cng nh cht lng keo rong c chit rt t loi rong .

Sn lng v ngun li rong bin trn th gii c th hin trn bng sau. Bng 1.1: Ngun li, sn lng thu hoch v tim nng sn xut rong bin [1]. (n v: 1.000 tn) Rong Rong Nu Khu vc Sn lng Sn lng Ngun li Ngun li thu hoch thu hoch Bc cc Ty bc i Ty dng ng bc i Ty dng Trung tm ty i Ty dng Trung tm ng i Ty dng a Trung Hi v bin en Ty nam i Ty dng ng nam i Ty dng Ty n dng ng n dng Ty bc Thi Bnh dng ng bc Thi Bnh dng Trung tm ty Thi Bnh dng Trung tm ng Thi Bnh dng Ty nam Thi Bnh dng ng nam Thi Bnh dng Nam cc Tng cng 35 72 10 50 23 7 4 3 545 20 7 1 30 807 100 150 10 50 1.000 100 100 120 100 650 10 100 50 20 100 2.660 6 223 1 1 1 75 13 5 10 822 1 153 1 1 1.315 500 2.000 1.000 150 50 2.000 100 150 500 1.500 1.500 50 3.500 100 1.500 14.600

b. Ngun li rong bin Vit Nam nc ta c khong 794 loi rong bin, phn b vng bin min Bc 310 loi, min Nam 484 loi, 156 loi tm thy c 2 min (Nguyn Hu Dinh, 1998). Trong c cc i tng quan trng l: rong Cu (Gracilaria), rong M

(Sargassum), rong ng (Hypnea), rong Mt (Porohyza) v rong Bn (Enteromorpha). Ngun rong trng bao gm ch yu cc loi rong nh: rong Cu ch vng (G.verrucosa), rong Cu cc (G.acerosa), rong Cu (G.asiatica v G.heteroclada), rong Sn (Alvarezii). Trong G.verrucosa v G.asiatica c trng vng nc l (Blackish water) t nm 1970 pha Bc, pha Nam t 1980 vi tng din tch 1.000 ha t sn lng khong 1.500 n 2.000 tn kh/nm. Rong Cu cc (G.acerosa) cng c trng vng thy triu, vnh, ao, a vi din tch khong 100 ha, sn lng khong 150 n 200 tn kh/nm. Rong Sn kapaphycus alvarezii c di trng vo vng bin nc ta nm 1993, loi rong ny c cht lng tt sn xut Carrageenan. Ngy nay trong nui trng Thy sn, cc nh nghin cu ang kt hp vic nui trng rong bin vi cc loi thy sn tm, c, nhuyn th nng cao hiu qu kinh t trn mt n v din tch mt nc. y l phng php hu hiu va pht trin nui thy sn va pht trin ngun li rong bin cc nc trn th gii ni chung v nc ta ni ring. Bng 1.2: Din tch cc vng c th qui hoch nui trng rong bin mt s tnh duyn hi Vit Nam [1]. (n v: ha) Din tch cc vng c th qui hoch nui trng Vng qui hoch Rong Cu cc Rong Sn Bnh nh Ph Yn Khnh Ha Ninh Thun Bnh Thun B Ra Vng Tu Tng cng 1.140 1.750 5.050 390 Cha qui hoch 4.500 12.824 ha 610 2.580 8.160 2.030 Cha qui hoch 6.140 19.520 ha

1.1.3 Phn loi rong bin Ty thuc vo thnh phn cu to, thnh phn sc t, c im hnh thi, c im sinh sn m rong bin c chia thnh 9 ngnh sau y: 1. Ngnh rong Lc (Chlorophyta) 2. Ngnh rong Trn (Englenophyta) 3. Ngnh rong Gip (Pyrophyta) 4. Ngnh rong Khu (Bacillareonphyta) 5. Ngnh rong Kim (Chrysophyta) 6. Ngnh rong Vng (Xantophyta) 7. Ngnh rong Nu (Phacophyta) 8. Ngnh rong (Rhodophyta) 9. Ngnh rong Lam (Cyanophyta) Trong , ba ngnh c gi tr kinh t cao l rong Lc, rong Nu, rong . * Ngnh rong Lc: c trn di 360 chi v hn 5.700 loi, phn ln sng trong nc ngt, nt c trng ca loi rong ny l c mu lc, sn phm quang hp l tinh bt. Rong c dng t bo n gin hoc phc tp, nhiu t bo dng hnh phin hay dng si, chia nhnh hoc khng chia nhnh. Tr mt s trng hp rong ch l mt t bo trn khng c v cn i a s c v ring l cht pectin hay Cellulose. * Ngnh rong Nu: c trn 190 chi, hn 900 loi, phn ln sng bin, s chi, loi tm thy trong nc ngt khng nhiu lm. Rong c cu to nhiu t bo dng mng gi, dng phin, dng si n gin, mt hng t bo chia nhnh, dng nhnh hoc phn nhnh phc tp hn thnh dng cy c gc, r, thn, l. Rong sinh trng nh (apical), gia, gc, cc lng. Ngoi ra, do cc t bo rong dng phin chia ct sinh trng khuch tn gi l sinh trng b mt. * Ngnh rong : Rong hay to l nhng loi rong bin khi ti c mu hng lc, hng tm, hng nu. Khi kh ty theo phng php s ch chuyn sang mu nu hay nu vng n vng. Trn th gii rong c s dng vi khi lng ln phc v con ngi, mt s loi c hm lng cao v Agar, Carrageenan, Fucellaran c s dng ch bin keo rong.

Phn ln rong c cu trc a bo, rong Gracilaria Verrucosa c di

nht. Tuy vy chiu di ca rong u ngn hn nhiu so vi rong Nu. Rong c 2.500 loi, gm 400 chi, thuc nhiu h, phn ln sng bin, c cu to t nhiu t bo, tr mt s t thuc dng mt t bo hay qun th. Rong c dng hnh tr dp di, phin chia hoc khng chia nhnh. Phn ln chia nhnh kiu mt trc (monopodial), mt s t theo kiu hp trc (symodial). Sinh trng ch yu nh, gia t hay phn tn. c trng ca loi ny l cha nhiu sc t , gia cc t bo c cha nhiu cht keo (Agar). Loi ny sinh trng vng bin su, ni thy triu thp, su t 6m n 7m. 1.1.4 Thnh phn sinh ha ca rong bin Theo Noris, Symeon, Williams trong 100 gam rong kh c [1]: m M ng : 24.5 g : 0.95 g : 31.0 g Provitamin A Vitamin B1 Vitamin B2 Acid nicotinic Vitamin C P : 1.23 g : 0.41 g : 2.07 g : 5.1 mg : 1.0 mg : 440 mg

Nhit lng : 30.3 Kcal Fe : 32 mg

Mui khong : 30.3 g

1.1.5 Gi tr dinh dng v ng dng ca rong bin. Gi tr dinh dng ca rong bin l cung cp y cc khong cht c bit l cc nguyn t vi lng, cc acid amin cn thit cho c th, cc loi vitamin, cc carbohydrate c trng v cc hot cht sinh hc c li cho c th, ng thi c kh nng phng v tr bnh. Theo s liu nghin cu ca Nht Bn trong rong Laminaria c cha cc vitamin sau y (miligam %): tin vitamin A (caroten)-1.1; A 622; B1 0.53; B2 0.41; acid nicotin- 1.6; acid folic 0.14; B12 0.0033 v ascorbic 28. Rong bin c hm lng lipid rt thp (t hn 2%). Nhng acid licozopentae kh cao ti 20 25% tng s lng cc acid bo, trong rong bin cn tm thy nhiu fucosterol v nhiu nguyn t vi lng khc. Trong rong bin c cha nhiu Iod. Iod hu c rt c gi tr trong y hc. Do vy rong Nu cn c dng lm thuc phng chng v cha bnh bu c (Basedow) [1]. Th d 1 Kg rong Laminaria cha mt lng Iod bng lng Iod c trong 100.000 lt nc bin.

Trong 10 gam rong kh loi Alginatearia esculenta cha mt lng vitamin E bng trong 100 gam c ci ng, trong 10 gam rong kh Gracilaria sn phm cha mt lng Canxi c trong mt cc sa [1]. Cc sn phm hu c t rong bin ngy nay c s dng ht sc rng ri trong cc ngnh nh: thc phm, dc phm, m phm, cng nghip dt, nng nghip, cng ngh sinh hc v nghin cu khoa hc. Cc polysaccharide t rong bin c coi l nhng hp cht hu c khng th thay th trong nhiu lnh vc cng nghip khi c s dng nh cht to ng, lm c, cht n nh v cht n nh. Gi tr cng nghip ca rong bin l cung cp cc cht keo rong quan trng nh: Agar, Alginate, Carrageenan, Fucryllanzan dng cho thc phm v nhiu ngnh cng nghip khc. Khong 20% sn lng rong bin th gii c dng sn xut cc loi keo rong, ch bin thc n cho vt nui v lm phn bn, s cn li ch yu c dng lm thc n cho ngi (Ohno v Critchley, 1997). Nm 1870 ngi ta iu ch x phng t cc cht K2O, Na2O ly t rong bin (rong Nu), pht hin trong rong Nu c cha Iod, t ngi ta dng nguyn liu rong Nu iu ch Iod [1]. Rong bin ngy cng c s dng rng ri lm thc phm. Nhu cu v rong lm thc phm Chu chim ti 90% ton th gii cn Chu u ch chim 1%. Tiu th rong nhiu nht l Nht, Nam Triu Tin, Trung Quc chim 70 90 nghn tn/nm, Bc M tiu th 240 tn/nm. Nh cc tnh cht vt l, rong c ch bin cng vi u, nhiu loi ng cc v rau qu khc thnh cc mn n c sc, dng t nhin hay qua s ch. Rong c s dng lm ph gia trong cc mn n ch bin t c, gip xc, nhuyn th, gi ch, ko bnh, ung. Porpyra cn gi l rong mt c gi tr thc phm cao nht v c cc nc phng ng rt a chung. Rong mt ti c a vo ch bin cc mn n khc nhau vi tht, c nh xo, nu canhhay nu ch gii kht. Rong c c tnh y hc: Bi b c th v khong cht, tng sc khng t nhin, iu ha s bin dng v cc ni tuyn, lm gim lng m, lm ti tr, chng lo ha, chng vim ng rut, chng bu, chng khp, kch thch s tun hon. Rong bin dng cha cc bnh thiu khong, bing n, vim thp khp,

10

bo ph, tim mch, thn kinh, bu cCc oligosaccharide c tc dng khng nm bnh, c ch cc qu trnh pht trin ca t bo ung th, t bo HIV, cc bnh thuc h tun hon cng nh h tiu ho ngy cng c pht trin trong nhiu loi rong bin. V vy ngy nay rong bin c xp vo loi thc phm chc nng v ngy cng c s dng rng ri khp th gii. Theo nghin cu rong Lc Nht Bn Laminaria c cha nhiu cellulose, cc glucid, acid Alginic, Fucoidin, cc mui ca acid Alginic. Tnh chng nhim x cao ca acid Alginic v cc mui ca n c s dng rng ri. Ngoi ra Fucoidin v Alginate ha tan trong nc cn lm gim cc tnh cht ng t trong mu v chng to ra cc khi u. Rong Laminaria cn l ngun cung cp cc acid amin asparagin v glutamine c c th con ngi hp th d dng. Trong cc t bo ca rong Laminaria cn tm thy cc acid amin ng ng c th Laminin c tc dng gim huyt p ca ng vt. 1.1.6 c im sinh trng, pht trin v thi k thu hoch rong bin hp l cho cng ngh ch bin. Qu trnh ln ln ca cy rong gm c nm thi k: Sinh trng pht trin tch ly sinh sn tn li. cui thi k tch ly, hm lng cc cht keo t cao nht v hon thin nht v cu trc ca n. Do cn thu hoch cui thi k ny l tt nht. Nu thu hoch rong non (cha tch ly ti a) hiu sut qui trnh gim, cht lng keo rong gim. Nu thu hoch rong tn li cng phm phi cc nhc im trn. C th xc nh c thi k ny theo cc ch tiu cm quan sau: Rong ang thi k phn nhnh cp II mnh, chiu di nhnh trn 20 cm i

vi rong hi t nhin v hn 30 cm i vi rong nui trng. Khi lng c th t t 2 n 3.5 gam. T l rong kh/ rong ti t 1/8 n 1/10. Vi rong Nu, thng quan st phao sinh sn ca n. Kh nng khai thc Mannitol tp trung cao vo thng 3 v 4.

11

Thi im khai thc tt nht i vi rong Nu sn xut Alginic l vo

cui thng 4 khi rong thnh thc sinh dc v phng thch giao t vo nc bin lu truyn ni ging cho th h sau. Nu khai thc sm hn (thng 3 chng hn) th nn bng cch ct cha li

gc 20 cm rong c th ti pht trin v hnh thnh c quan sinh sn. Qua kt qu phn tch hm lng cc cht axit Alginic, Mannitol v Iod trong cc loi rong Nu tm thy vng bin Qung Nam Nng n Ninh Thun (Trn Th Luyn, Nguyn Anh Tun v cng tc vin, 1998) cho thy: Hm lng axit Alginic tch ly trong cc loi rong khc nhau c s sai khc

nhau r rt. Hm lng axit Alginic cao nht thng vo thng 4 l lc cc loi rong trng thnh, phng thch giao t v kch thc ca cy rong l ln nht sau t thng 5 tr i th c hin tng hm lng axit Alginic gim dn theo s tn li ca rong. Hm lng axit Alginic trong 5 loi rong Nu dao ng trung bnh t 23.5

37.05% trng lng kh tuyt i. Hm lng ny cao nht vo thng 4, ng vo thi k rong trng thnh. Hm lng Mannitol cng bin i tng t nh axit Alginic,cao dn theo s

sinh trng ca rong v cng t gi tr cao nht lc rong trng thnh, phng thch giao t vo thng 4. Hm lng Mannitol trong 5 loi rong Nu vng bin Khnh Ha c phn

tch, c hm lng trung bnh vo khong 6.3 11.35% trng lng rong kh tuyt i. Trong loi S.mcclurei c hm lng ln hn c. Hm lng ny thng cao vo khong thng 3, 4. Hm lng Iod trong cc loi rong Nu c phn tch c dao ng trung bnh

t 0.07 0.09% trng lng kh tuyt i. Trong loi Turbinaria ornate v loi S.kjellmanianum c hm lng cao. Thng hm lng Iod tp trung cao vo cc thng ma ng cn cc thng ma h th hm lng lng ny thp. Cc loi rong c hm lng axit Alginic v Mannitol cao l Sargassum

mcclurei, Sargassum kjellmanianum, Sargassum quinhoneness Dai, Sargassum

12

polycystum. Cao nht l loi Turbinaria ornate. Trong loi Sargassum mcclurei c tr lng ln nht, v Turbinaria ornate c tr lng khng ng k. T cho thy nn khai thc rong vo thng 4 v u thng 5 l lc rong Nu trng thnh, c kch thc ln nht, hm lng axit Alginic v Mannitol cao nht. Mt khc lc ny rong phng thch cc giao t vo nc bin do vy ta va thu va bo v c li ch lu di, trnh c nn hy dit ngun li. 1.1.7 Tnh hnh s dng, ch bin rong bin Vit Nam. Mc d c thin nhin u i v bin nhng s u t pht trin nui trng, ch bin khai thc rong bin cn hn ch v cha c hiu qu. Ngnh cng nghip ch bin rong bin cha pht trin, hin nay ch mi c nh my c hp H Long Hi Phng sn xut vi cng sut nh. Nm 1985 B Thy sn xut khu c 150 tn v nm 1986 c 100 tn rong kh cho Nht Bn. Nhu cu Alginate v Agar ngy cng tng, c nhiu c s cng nghip phi mua Alginate ca Nht Bn vi gi kh cao. Trong thi gian ti nn cng nghip cng pht trin th nhu cu v Alginate, Agar v cc keo rong khc s cn tng gp bi. Nu c u t v pht trin ng mc, cng ngh rong bin s mang li hiu qu ln cho nn kinh t nc nh. Rong bin l ngun nguyn liu qu, c kh nng gip cho c th phng chng c mt s loi bnh. Do vy nhiu nc trn th gii ginh khon ngn sch kh ln cho vic nghin cu ng dng sn xut thc phm t rong bin. nc ta, thc phm t rong bin cha thc s c ch , l mt vn cn ang b ng. Mt s c s ch bin nh ti gia nh nh: lm gi, nu thch, ng sng, mt, ko, ch rong bin,Tuy nhin cc sn phm ny cha nhiu, cha ph bin, rt t ngi dn bit n cc loi thc phm c bit ny. Do cn phi c k hoch pht trin mnh hn tin ti cc sn phm rong bin phi phong ph hn, c sn xut theo quy m cng nghip trong iu kin v sinh am ton thc phm cao phc v tiu dng ni a v xut khu. 1.2 Qu trnh vn chuyn v cc bin php bo qun rong kh. Trong qu trnh thu hi, ch bin v thng mi rong bin thng phi vn chuyn rong bin vi khi lng kh ln. Trong qu trnh vn chuyn rong kh

13

thng ng rong thnh kin tng cng kho cha v phng tin vn chuyn. Kin rong phi c ng gi bng dy ai dc, ngang c gn nhn hiu ghi tn sn phm, c s sn xut, khi lng tnh. Trong khi ch i, rong phi c trn cc sn g cch mt t, cch tng 20 cm. Hoc c th ct rong thnh cc mnh c chiu di 3 n 5 cm ri ng gi vo bao ti hoc bao nylon. Vn chuyn rong bng cc phng tin c mi che. Phng tin phi kh sch, khng vn chuyn rong kh cng vi cc loi hng ha ti sng, t nh tm, c, Mt s hin tng h hng ca rong: Trng thi cy rong b thay i: rong gin, mn. Gin l do sy nhit kh

cao, rong mn l do s ch nc ngt khng ng k thut, hm lng mui cn nhiu. Cc loi vi sinh vt nh cellulomonas, Aspegillus, Streptococcus, Pseudomonas v Penicilium hot ng mnh phn hy cellulose v cc cht keo rong. Rong hao ht trng lng do m cao. Rong h cc b: do tri rong xung nn nh m khng tn nhit, xut hin s

t pht nhit nm mc pht trin. Cc bin php bo qun rong kh. Rong cha phi thng thong, lu thng khng kh. Khng kh trong kho c

m 80%. Ngy kh ro phi m ca kho gim m ca kho. Cc kin rong c trn cc gin cch mt t 15 20 cm. Gia cc gin

c li i li thng xuyn kim tra, bc xp, to thong tn nhit. Pht hin rong m phi a i ch bin ngay. Khi rong mc phi loi b

phn mc, ra, sy li. Cc kin rong phi c sp xp theo cht lng v thi gian sn xut, rong

nhp kho trc phi a sn xut trc. Rong kh ng tiu chun, bo qun ng ch thi gian ti a l 1 nm. 1.3 Gii thiu v rong Nu v rong M (Sargassum).

1.3.1 c im. Ngnh rong Nu (phaeophyta) c khong 190 loi, trn 900 loi phn ln l sng bin, s ging loi tm thy trong nc l v nc ngt khng nhiu lm.

14

Rong M Sargassum l mt ging To ln thuc h rong M Sargassaceae ca ngnh rong Nu sng tri ni trong nc. Thn cy c dng tr gn trn rt ging vi thc vt bc cao c mu t xanh oliu n nu ( mu nu m, nu vng). Mu sc khc nhau cc ging loi ph thuc vo thnh phn v t l ca cc loi sc t c trong rong. i vi rong c lm kh s c mu nu m [2], [19]. Thn rong di ngn ty loi v ty iu kin mi trng. Thng gp t vi chc cm n vi ba mt hay hn (6- 8 m). Rong m l loi rong to mc thnh bi, gm vi trc chnh quanh nhnh, nhnh mang phin c dng ca l, phin c rng mn ging nh l m do c tn l rong l m hay gi tt l rong M. Cc loi rong M u c phao, phao nhiu t to nh khc nhau, hnh dng ca phao l hnh cu hay tri xoan, ng knh ca phao nh khong 0.5 0.8 mm, phao ln khong 5 10 mm. Phao c th mang cnh hoc khng. Rong l m l nhng loi rong mc nhng vng bin m nng, trn nn vi, san h cht, ni sng mnh v nc trong, nht l ven cc o.Chng mc t pha trn ca mc nc trung bnh thp ca con nc thng n su t 2 4 m. Sinh trng trung bnh t 2.000 4000 b/m2, c ni n 7.000 g/m2 nh Hn Chng, Nha Trang. Rong M l loi c kch thc c th ln v tr lng cao nht trong cc loi rong bin Vit Nam. Rong M mc trn tt c cc loi vt bm cng, trn cc vch dc ng, cc bi tng, cc vng c ngm hay rn san h ngm, nhng thch nghi nht l trn vt bm san h. Cu trc ca cc qun th rong M trn cc vt bm khc nhau r rt. Trn cc b dc ng, chng phn b thnh ai hp di mc triu thp n su khong 0.5 m. cc b bin tng nm trn nn ct hay cui, chng mc thnh qun th dy, phn b tng i u, mt khi rong trng thnh c th t 10 c th/dm2, cho nn vo ma pht trin ca chng rt t cc loi rong bin khc c th mc chen c vo trong qun th rong ny. a s cc loi rong u thch mc ni c sng mnh. cc o, b pha ng chng mc dy v phong ph hn b pha Ty. cc bi hng ra bin khi, chng pht trin mnh v sinh lng cao hn nhiu so vi cc bi rong trong

15

cc vng, vnh yn sng. Cc bi rong trn b bin dc, thm san h cht, vi ng vai tr quan trng trong ngun li ca rong M, nhiu vng rng 30 50 ha hay hng trm ha, ko di vi chc km, thng gp ven bin min Trung, nht l t Qung Ngi n Bnh Thun. Ma v rong M c s sai khc cht t ty thuc tng loi, ni phn b, ty cc iu kin mi trng sng,nhng nhn chung qui lut v ma v kh r rt. Chng tng trng rt mnh t thng 2 n thng 3, a s cc loi c kch thc ti a vo thng 3, 4 v hnh thnh cc c quan sinh sn, sau s b sng nh tp vo b v tn li. n thng 7 hu ht cc loi rong u tr tri. Mt s loi mc ln cao hoc phn b ln cao (vng triu thp) nh: S.mcclurei, S.polystum pht trin v tn li sm (thng 4). Trong khi cc loi mc vng di triu nh S.binderi, S.microcystum,mc chm hn, n thng 6, 7 i ni vn cn cc qun th rong ny. Mt vi loi thch nghi trong cc vng, vnh yn sng c th tn ti v pht trin tt vo thng 7 nh S.polycystum v S.longicaulis. Rong trng thnh v phng thch giao t vo cc thng 3, 4, 5, vo thi im ny, kch thc ca rong t n ti a v sinh lng cao nht. Mt khc hm lng axit Alginic cng cao nht. Cc c im ny rt quan trng, ph hp v c li cho vic khai thc ngun li t thng 4 tr i. Vic khai thc ng ma v hon ton c kh nng bo v ngun ging t nhin, gip cho rong ti pht trin li vo ma sau. Ngoi ra vic khai thc bng cch ct gc rong tng 10 cm gip cho mt s nhnh cn st li vn tip tc pht trin to ra cc c quan sinh sn. Cc bi rong M mc trn thm san h cht c din tch rng ln, mt dy, sinh lng cao (trn 12 kg rong ti/m2) rt quan trng vi ngun li, tm thy cc tnh Qung Ninh, Qung Ngi, Bnh nh, Khnh Ha, Ninh Thun v Bnh Thun. Cc bi rong rng ln nm gn trc giao thng rt thun li cho vic khai thc v vn chuyn. Sn lng hng nm c tnh c th t 10.000 tn rong ti [1].

16

1.3.2 S phn b [1]. Bng 1.3: Cc ging loi rong Nu tm thy v phn b.
a phng STT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Loi rong Sargassum mcclurei Sargassum graminafolium (rong M tro l nhnh) Sargassum phamhoangii (mt loi rong mi t m thy Vit Nam) Sargassum siliquosum Padina australis (rong cnh qut) Sargassum crassifolium Sargassum patens var. Vietnamese Dai Sargassum quinhonense Dai Sargassum polycystum Sargassum kjellmanianum Sargassum microcystum Sargassum congkinhii Turbinaria ornate (rong ci bp) Padina tetrastromatica (rong qut 4 lp) Q. Nam Nng Bnh nh Khnh Ha Ninh Thun

Rong Nu phn b ti vng bin Qung Nam Nng khng nhiu so vi vng bin Khnh Ha v Ninh Thun. Qung Nam Nng tuy c nhiu trin dc, bi ci, bi san h cht nhng c chiu ngang rt hp (1 n 10 m) nn din tch phn b rt nh, tr lng kh cao. Khi lng rong ti trung bnh q0 = 2 4 kg/m2, c bit c ni t n 7 kg/m2 nh vng C Lao Chm, trin o Hi Vn. Din tch rong M mc ti ch vng bin Qung Nam Nng khong 190.000 m2 , tr lng rong mc ti ch c th thu c vo thng 4 khong hn 800 tn rong ti. y l kt qu khng ln i vi mt vng bin c iu kin t nhin thun li cho s pht trin ca rong M. Kt qu ny rt nh so vi cc va rong

17

M khc nm ri rc ven bin min Trung. Rong t kch thc v sinh lng cao nht vo cui thng 3 v u thng 4. Sau rong sng cm c thm mt thi gian na ri tn li vo thng 7. Din tch c rong mc ti ch ca tnh Bnh nh khong hn 40.000m2 , tr lng rong ti c tnh hn 100 tn/nm. Sinh lng cao nht vo cui thng 4 v u thng 5. Din tch c rong phn b rt b so vi cc tnh khc, sinh lng trung bnh q0 khong 2.5 kg/m2. Cc vng c rong mc l Bi Xp, Ghnh Rang, Hn Kh.Trong vng Hn Kh l vng c chiu di bi rong khong 10 km, rong mc khng u di rong hp, c tr lng cao nht. Tr lng rong ca tnh Bnh nh thp nht trong cc tnh iu tra. Hin nay s rong ny hng nm t mc, t tn li, khng c k hoch no khai thc s dng, cn rt lng ph. Vng bin Khnh Ha l vng c din tch rong M mc cao nht trong cc tnh iu tra, tng din tch c rong ln ti 2.000.000m2, tr lng c th khai thc c hng nm c th c tnh hn 11.000 tn rong ti. Khnh Ha c nhiu vng rong nh Hn Chng, Bi Tin, bn o Cam Ranh, Hn Tre v mt s o khc. Bng 1.4: Tr lng rong M theo vng bin cc tnh.
STT A IM DIN TCH (M2) THNG TR LNG RONG TI (tn)

1 2

Qung Nam Nng Bnh nh

190.000 42.750

4/97 3/97 4/97 5/97 3/97 4/97 5/97 3/97 4/97 5/97 3/97 4/97 5/97

860 106 124 129 11.002 7.930 6.046 7.650 6.180 4.650 18.750 15.724 10.825

Khnh Ha

2.000.000

Ninh Thun

1.500.000

Tng cng

3.732.750

18

1.3.3 Thnh phn ha hc ca rong Nu. a. Sc t Sc t trong rong Nu l dip lc t (Chlorophyl), dip hong t (Xantophyl), sc t mu nu (Fucoxanthin), sc t (Caroten). Ty theo t l cc loi sc t m rong c mu t nu vng nu nu m vng lc. Nhn chung sc t ca rong Nu kh bn [1]. b. Gluxit Monosacaride [1]. Monosacaride quan trng trong rong Nu l ng Mannitol c Stenhouds pht hin ra nm 1884 v c Kylin (1913) chng minh thm. Mannitol c cng thc tng qut: HOCH2 (CHOH)4 CH2OH. Mannitol tan c trong Alcol, d tan trong nc c v ngt. Hm lng t 14% 25% trng lng rong kh ty thuc vo hon cnh a l ni sinh sng. Hm lng Mannitol trong 2 loi rong Nu c cc nh nghin cu Vin Hi Dng hc Nha Trang xc nh trn bng 1.5. Bng 1.5: Hm lng Mannitol 2 loi rong S.mcclurei v S.kjellmanianum ti Hn Chng Nha Trang 1979 (% so vi trng lng kh tuyt i) Loi rong Hm lng Mannitol trung bnh (%) ca thng 1 2 3 4 5 6 Thng 6 rong bt u tn li Ghi ch

S.mcclurei

11.3

14.6

15.79

16.73

16.36

9.37

S.kjellmanianum 10.12

10.53

12.40

13.82

6.95

Thng 5 rong bt u tn li

Hm lng Mannitol bin ng theo thi gian sinh trng trong nm ca rong kh r rt, tng dn t thng 1, tp trung cao vo ma h (thng 4) ri sau gim i: Theo Kylin (1993) v Vedrinski (1938) cho thy hm lng Mannitol t

19

25% v ma h ri b phn hy dn trong cc thng ma ng ch cn 4% 6%. Vit Nam, theo kt qu nghin cu ca cc nh khoa hc vin Hi Dng hc (1979) v i hc Thy sn (1997 1998) hm lng Mannitol cng theo quy lut ny. Thng 4 l hm lng Mannitol cao hn c, Mannitol trong loi S.mcclurei c hm lng cao nht, vo thng 4 t 15.9% so vi trng lng kh. Hm lng Mannitol ca cc loi rong cc vng bin Qung Nam Nng, Bnh nh, Khnh Ha, Ninh Thun dao ng t 7% 15.95% (vo thng 4 hng nm). Trong qu trnh bo qun rong kh c hin tng xut hin cc im m trng trn thn cy rong, l hn hp ng Mannitol theo t l : mui 60% 80%, Mannitol 20% 40%. Rong bo qun khng tt, m cao lm cho Mannitol b ph hy. Cng dng ca Mannitol: dng trong y hc cha bnh cho ngi gi yu; trong quc phng dng iu ch thuc n theo t l hn hp Mannitol vi Hydrogen v Nito. Ngoi ra Mannitol cn dng iu ch thuc st trng (Mannit vi kim loi c tc dng st trng cao). Do c kh nng ny m ngy nay mt s tc gi cho thy c kh nng sn xut thuc tr su c bn cht sinh hc (bo v thc vt) t rong bin. Polysaccharide Alginic: Alginic l mt polysaccharide tp trung gia vch t bo, l thnh

phn ch yu to thnh tng bn ngoi ca mng t bo rong Nu. Alginic v cc mui ca n c nhiu cng dng trong ngnh cng nghip, y hc, nng hc v thc phm. Hm lng Alginic trong cc loi rong Nu khong 2% 4% so vi rong ti v 13% 15% so vi rong kh. Hm lng ny ph thuc vo loi rong v v tr a l mi trng m rong sinh sng. Theo cc ti liu tng kt ca Miyake (1995) cho thy hm lng Alginic trong cc loi rong Nu c cc vng bin Lin X c l 13% 40%. Theo ti liu phn tch cc chuyn gia B Thy sn cho thy hm lng Alginic trong cc loi rong Nu Hi Phng l 22% 40%, trong khi , rong Nu vng bin Ph Yn Khnh Ha c hm lng Alginic cao hn hn. Theo s liu phn tch ca cc nh nghin cu vin Hi Dng hc th hm lng Alginic ca hai loi rong Nu c trnh by trn bng 1.6.

20

Bng 1.6: Hm lng axit Alginic trong cc loi rong Nu (% so vi rong kh tuyt i). Loi rong Thi gian thu mu (thng/nm) 11/77 4/78 1/79 6/79 5/1978 11/77 4/78 1/79 6/79 3/1978 11/77 4/78 3/1978 5/78 4/79 3/1978 3 4/1979 a im thu mu Phm vi dao ng ca hm lng 26.41 43.36 29.89 39.30 40.06 31.60 43.64 37.64 44.77 42.747 35.17 41.00 34.82 39.11 43.50 39.60 36.41 41.93 Hm lng trung bnh 33.24 35.61 37.03 40.39 38.47 41.30 39.17 43.75 l kh cao,

S.mcclurei Hn Chng Sn Hi S.kjellmanianum Hn Chng S.polycystum S.congkinhii Hn Chng Sn Hi S.microcystum Hn Chng S.feidmanii Sn Hi Turbinaria 3 5/1979 39.14 47.16 ornata Hm lng Alginic c cc loi rong min Trung Vit Nam

dao ng t 12.3% 35.9% so vi trng lng kh tuyt i, ty thuc vo loi v vng a l. Trong loi rong S.mcclurei v Turbinaria ornate c hm lng Alginic cao nht khong 35.9% 39.4% rong kh tuyt i. Nu so snh cc vng bin th rong bin vng bin Khnh Ha c hm lng Alginic cao hn c (t 26.2% 39.4% rong kh tuyt i) [1]. Axit Fucxinic: c tnh cht gn ging vi axit Alginic. Axit Fucxinic tc dng vi axit Sunfuric to hp cht c mu ph thuc vo nng axit Sunfuric. Chng hn: Axit Fucxinic + H2SO4 0.1% : cho sn phm mu xanh. Axit Fucxinic + H2SO4 10% : cho sn phm mu xanh tm Axit Fucxinic + H2SO4 25% : cho sn phm mu tm Axit Fucxinic + H2SO4 25% : cho sn phm mu Axit Fucxinic + H2SO4 > 50% : cho sn phm mt mu

21

Nh c tnh cht ny m Fucxinic c ng dng vo sn xut t si mu, phim nh mu. Mui ca Fucxinic vi kim loi gi l Fucxin. Fucxinic tc dng vi Iod cho sn phm mu xanh [1]. Fuccoidin: L loi mui gia axit Fuccoidinic vi cc kim loi ha tr khc nhau nh: Ca, Cu, Zn. Fuccoidin c tnh cht gn ging vi Alginic, nhng hm lng thp hn Alginic. Fuccoidin hay Fucoidan l mt polysaccharide sulfate ha d hp, trong fucose chim t 18.6% 60% [18], sulfate chim t 17.7% 39.2%, ngoi ra cn c mt cc thnh phn ng khc nh: galactose, glucose, mannose, xylose, rhamnose, v acid uronic. Hm lng fucoidan trong cc loi rong Nu khong 2 9% so vi rong kh. Hm lng ny ph thuc vo loi rong v v tr a l mi trng m rong sinh sng [18]. Laminarin: Laminarin l tinh bt ca rong Nu. Laminarin thng dng bt khng mu, khng mi v c hai loi: loi ha tan v loi khng ha tan trong nc. Laminarin c hm lng t 10% 15% trng lng rong kh ty thuc vo loi rong, v tr a l v mi trng sinh sng ca rong Nu. Thng th vo ma h hm lng Laminarin gim v phi tiu hao cho qu trnh sinh trng v pht trin ca cy rong. Cng dng ca Laminarin l dng cho thc phm v chn nui [1]. Laminarin c hnh thnh t cc gc D- glucans kt hp vi nhau bng cc lin kt - 13 v mt t lin kt - 16, gc ng cui mch ca mt s phn t c th c cc gc manitol (M series) hoc vn l glucose (G series) [17]. Cellulose: l thnh phn to nn v cy rong. Hm lng Cellulose trong rong Nu nhiu hn rong . Cng dng: Dng cho cng nghip giy, trong cng nghip xy dng (l ph gia kt cu xi mng) [1]. c. Protein Protein ca rong Nu khng cao lm nhng kh hon ho. Do vy rong Nu c th s dng lm thc phm. Protein ca rong Nu thng dng kt hp vi Iod to Iod hu c nh: MonoIodInzodizin, DiIodInzodizin. Iod hu c rt c gi tr trong y hc. Do vy rong Nu cn c dng lm thuc phng chng v cha bnh bu c (Basedow). Hm lng protein rong Nu vng bin Nha Trang dao ng t 8.05% 21.11% so vi trng lng rong kh. Hm lng cc axit amin cng ng k v c gi tr cao trong Protein ca rong bin. Theo ti liu nghin cu ca

22

chuyn gia rong bin quc t, hm lng axit amin mt s loi rong Nu vng bin Jeddah, Saudi Arabia c trnh by trn bng 1.7 [1]. Bng 1.7: Hm lng axit amin mt s loi rong Nu vung bin Jeddah, Saudi Arabia (mg %). Axit amin (mg %) Asp Thr Ser Glu Pro Gly Ala Val Cys Met Tre Leu Tyr S.vulgare 12.70 3.23 5.90 16.50 Nd 6.20 5.78 4.77 0.72 0.53 4.06 7.98 2.56 S.enerve 12.86 3.25 6.60 18.03 Nd 6.91 5.91 5.21 0.66 0.76 4.13 8.08 2.67 C.barbata 13.3 1.7 3.4 24.2 2.7 1.4 1.3 1.5 2.5 0 4.9 6.3 5.7 T.murrayana P.vikersiae 12.2 2.2 3.6 21.2 3.6 1.3 1.5 1.4 2.9 0 4.7 7.6 6.2 12.4 3.4 6.7 15.1 2.9 2.7 3.0 0.3 5.2 0.7 5.6 9.1 3.5

Nd = not determined (khng xc nh) d. Hn hp Phenolic Theo Ragan v Glombitza, hp cht rt ph bin trong rong Nu thuc nhm hp cht polyphenolic l phlorotannin. Phlorotannin l cht chuyn ha th cp, xut hin ch yu cc m, ti nng c th ti 20% so vi khi lng tnh ca rong bin. Schoenwaelder 2002, ch ra mt s chc nng ca chng nh tng tnh lin kt v chc cho thnh t bo. Phlorotannin hp th bc sng UV, ch yu l UVC v mt phn UVB, vi cc i ti 195 nm v 265 nm [9]. e. Cht khong Hm lng cc nguyn t khong trong rong Nu thng ln hn nc bin. Chng hn: I2 ca rong Nu ln hn trong nc bin t 80 n 90 ln. Hm lng

23

Ba ln hn trong nc bin 1.800 ln. Mt s loi rong Nu cn c kh nng hp th mt s cht phng x, do vy c th dng rong Nu xc nh nhim phng x ca mt vng a l no . Hm lng khong ca cc loi rong Nu Nha Trang dao ng t 15.51 46.30% ph thuc vo ma v, thi k sinh trng. Chiu hng tch ly khong theo Wort cho thy lng khong ti a v ma ng. Theo s liu ca cc nh nghin cu Vin Hi Dng hc cng nhn xt theo quy lut . Chng hn hm lng khong tng s ca loi S.mcclirei bin ng theo ma r rt cao nht vo thng 12/1977 l 42.805% ri li gim dn v thp nht vo thng 3/1978 l 21.1% sau li tng dn vo thng 7/1978 l 33.75%. Bng 1.8: Hm lng Iod trong cc loi rong Nu (% trng lng kh) Nha Trang Khnh Ha. Thi gian thu mu 11/77 4/78 1 6/1979 S. Kjellmanianum 11/77 4/78 1 5/1979 11/78 5/1979 5/1979 a im ly mu Hn Chng ---Sn Hi -Phm vi dao ng ca hm lng 0.06 1.13 0.05 0.08 0.05 0.15 0.05 0.11 0.05 0.11 0.1 Hm lng trung bnh 0.09 0.07 0.09 0.07 0.08 --

Loi rong S. mcclurei

S.polycystum S. congkinhii

S.feldmanii 3 4/1979 -0.12 0.13 -Turbinaria 3 5/1979 -0.1 0.16 0.15 ornata Hm lng Iod trong mt s loi rong Nu dao ng 0.05% 0.16% so vi rong kh tuyt i. Hm lng ny c phn thp hn rong Nu vng bin Hi Phng trung bnh trong khong 0.25% 0.34% v cng thp hn rong Nu vng Vin ng (pha Bc Lin X c) c hm lng trung bnh 0.24%. S bin i hm lng Iod kh r rt, thng vo ma ng rong Nu c hm lng Iod cao hn ma h.

24

1.4

Mt s quy trnh cng ngh sn xut cc cht t rong Nu.

1.4.1 Cng ngh ch bin Mannitol t rong Nu [1].

Rong ti X l ct nh L= 23 cm Sy s b W = 30%, T0 = 50 600C Alcol thu hi Chit Mannitol T0C = 80 850C Phn tch Rong kh

X l ct nh L = 2 3 cm

Alcol 960 T l v/w = 56/1 rong

Dch chit

B rong

Lc li

Chng ui alcol

Chng ui dung mi C c, w = 50%

Sn xut alginate

Kt tinh, T = 2 40C
0

(5 6 ln)

Ha tan

C c

Kt tinh

Lm kh

Bao gi

25

Hnh1.1: S quy trnh chit Mannitol bng phng php ha tan trong Alcol 1.4.2 Chit rt Iod t rong Nu [1].
Rong Nu

Ra sch

X l qua formol

Ra

Ct nh (1 1.5 cm)

Chit Iod

Nc 800C, pH = 7. Thi gian: 10 15 pht

Lc tch

Dch chit 3 ln

B rong

C c

Ch bin alginate

Cao rong

Cht bo qun

nh hnh

Sy kh

Bao gi

Hnh 1.2: S quy trnh tng qut chit rt Iod t rong Nu.

26

1.5

Tng quan v hp cht Phlorotannin (polyphenol).

1.5.1 c im. Theo Ragan M. A [9] th phlorotannin c nh ngha nh sau: chng l polyme sinh hc c hnh thnh t gc phloroglucinol (1,3,5 trihydroxybenzen) v cc gc ny lin kt vi nhau bng nhiu cch khc nhau. Chnh v vy m chng to nn nhiu cu trc khc nhau v i lc c trng lng phn t ln n 650 KDA. Cho n nay, trn th gii c mt s cu trc ca cc hp cht thuc nhm phlorotannin t rong Nu c xc nh v nhiu dch chit th ca chng c kho st hot tnh sinh hc. Tuy nhin, rt t cng trnh cng b mt cch y cc cu trc ca tt c cc dn xut ca phloroglucinol trong dch chit phlorotannin th c chit t mt loi rong c th v hot tnh ca chng v cc cng trnh ny ch c cng b trong 10 tr li y. Cc nh khoa hc Hn Quc [10] cng a ra cu trc y ca cc dn xut ca phloroglucinol chit t loi rong Ecklonia cava bao gm:

27

Hnh 1.3: Phloroglucinol (i) v phlorotannin [tetrafucol A (ii), fucodiphloroethol B (iii), fucodiphlorethol A (iv), tetrafuhalol A (v), tetraisofuhalol (vi), phlorofucofuroeckol (vii)] v hot tnh ca chng. 1.5.2 C ch oxi ha ca polyphenol. Trong c th con ngi thng xuyn din ra nhiu hot ng hoc xy dng hoc ph hy. C nhng cht tng nh l thc phm chnh ca t bo nhng ng thi cng lm hi t bo. C nhng phn t gy ra tn thng th cng c nhng cht khng li hot ng ph phch ny. Gc t do, oxygen v cht chng oxi ha l mt v d. Theo cc nh khoa hc th gc t do c th l th phm gy ra ti hn 60 bnh, Chng tn cng ln cc phn t protein, lipid, ADN,dn n cc bnh nan y nh thoi ha thn kinh, ung th, tim mch, tiu ng, V vy ch n nhiu hoa qu v t cholesterol s lm gim ung th, km hm v gim cc gc t do. Cc cht chng oxi ha t tri cy hoc sn phm chit xut gip bo v, ngn cn qu trnh oxi ha. chng li tc hi ln do cc gc t do gy ra, c th con ngi c trang b mt h thng cc cht chng oxi ha. Cht chng oxi ha l cc cht

28

c kh nng ngn nga, chng li v loi b tc dng c hi ca cc gc t do trc tip hoc gin tip. Cht chng oxi ha c th phn ng trc tip vi gc t do hot ng, to ra nhng gc mi km hot ng hn, ngn cn cc chui phn ng dy chuyn do cc gc t do khi mo. Cht chng oxi ha cng c th to phc gin tip vi ion kim loi chuyn tip c ch enzim xc tc trong phn ng sinh gc t do nhm ngn s hnh thnh gc t do trong c th. Polyphenol c coi l cht chng oxi ha hu hiu nht hin nay (hu hiu gp 100 ln vitamin C v gp 25 ln vitamin E) [5]. S oxi ha ca cc hp cht polyphenol c ngha rt ln trong s sinh tn g hp nhiu cht t nhin nh licnhin, alkaloid v melanin. S to thnh cc hp cht ny c th gii thch bng c ch gc t do, trong giai on u ca hp cht monomerphenol xut hin cc gc phenol. giai on sau, xy ra s ghp i cc gc t do vi s to thnh mi lin kt C-C hoc C-O-C ph thuc vo kiu ghp i [5]. Kh nng khng oxi ha mnh ca polyphenol c gii thch da trn s lng ln cc nhm hydroxyl trong cng thc chng , n cho ghp tip nhn electron mt cch d dng (polyphenol l cht nhn cc gc t do), tc l chng c kh nng dp tt cc qu trnh to ra gc t do. 1.5.3 Hot tnh sinh hc ca phlorotannin. Hot tnh sinh hc ca phlorotannin c cng b nhiu nht l hot tnh chng oxi ha. Hot tnh ny c nghin cu thng qua kho st kh nng qut cc gc DPPH (2,2-diphenyl-1-picrylhydrasyl), gc hydroxyl (OH), cc anion nh superoxide anion (O2-), kh st (III), to phc chelat vi cc ion ha tr (II) v oxi ha lipid ca chng. Cc nh khoa hc Mexico [11] xc nh kh nng qut cc gc DPPH, anion O2- v hot tnh kh ca hn hp cc cht thuc nhm phlorotannin c chit t mt s loi rong Nu sinh trng ti vng bin nhit i ti Mexico. H thu cc phlorotannin bng chit lnh, s dng dung mi l dichloromethanol : methanol (2:1) trong 20 gi. Kt qu thu c cho thy tt c tng phlorotannin c chit t cc loi rong (Lobophora variegate, Padina gymnospora, Dictyota

29

cervicornis v mt s loi rong thuc chi Sargassum, chi Turbinaria) u c hot tnh chng oxi ha vi gi tr EC50 ca gc DPPH tng ng l nh sau: Lobophora variegate: 0.32 0.01 mg/ ml, Padina gymnospora: 3.45 mg/ml, chi Sargassum: t 6.64 7.14 mg/ml, chi Turbinaria: 8.85 mg/ml v chi Dictyota: 6.42 mg/ml. Ngoi ra tng phlorotannin chit t loi Lopophora variegate cn th hin kh nng qut gc anion O2- v tnh kh mnh. Hot tnh khng vi sinh vt ca phlorotannin chit t loi rong Ecklonia kurome ti Nht Bn c Koki Nagayama v CS kim nh [12]. th hot tnh khng vi sinh vt ca phlorotannin, nhm tc gi s dng phng php chit phlorotannin bng cn methanol. Hn hp thu c sau khi chit bng cn c chit li vi ethyl acetate v cui cng c tch trn sc k bn mng. Tt c cc phn on thu c u th hot tnh trn 25 chng vi sinh vt kim nh thuc 02 nhm Gr(+) v Gr(-). Kt qu l tt c cc phn on u th hin hot tnh khng 25 chng vi sinh vt em th. Trn c s phlorotannin th chit t loi rong Ecklonia cava cc nh khoa hc Hn Quc xc nh c nhiu hot tnh sinh hc khc ca chng. Tt c cc dn xut ca phloroglucinol phn lp c (tetrafucol A, fucodiphloroethol B, fucodiphlorethol A, tetrafuhalol A, tetraisofuhalol, phlorofucofuroeckol) u th hin hot tnh c ch cc enzyme Matrix metalloproteinase (MMP), Alphaglucosidase, alpha- amylase v c ch qu trnh gii phng histamine [13, 14, 15]. Kt qu ny nh hng cho vic s dng chng h tr iu tr cc bnh tiu ng, thp khp, vim nhim kinh nin, chng d ng. 1.5.4 ng dng. Polyphenol l nhng cht chng oxi ha, do chng c ng dng trong nhiu lnh vc. ng dng c quan tm mnh m nht hin nay l v kh nng dc l. Bn cnh kh nng ngn chn c hiu qu cc qu trnh oxy ha, c bit l qu trnh oxi ha lipid. Nhm cht ny c quan tm s dng trong cc sn phm thuc cc lnh vc khc nhau nh m phm, thc phm v dc phm.

30

Ring trong ngnh thc phm, do c im xut pht t thin nhin, m bo tnh an ton nn chng c s dng ch yu nh mt cht ph gia chng oxi ha, nhm thay th cho cc ph gia tng hp thng dng nh: BHA (Butyl hydroxyl anisol), BHT (Butyl hydroxyl toluene), nhng cht ny d gy tc dng ph khng c li cho sc khe ngi s dng. Polyphenol ch yu c ng dng lm bn cht bo trong cc sn phm bnh, ko, Ngoi ra, polyphenol (trong ch, rong bin, ) cn c s dng nh mt loi thc phm chc nng nhm tng cng kh nng phng v cha bnh ca c th [5]. Phlorotannin trong rong Nu cn c hot tnh khng khun [6], khng nm bnh, c ch cc qu trnh pht trin ca t bo ung th, t bo HIV, cc bnh thuc h tun hon nh iu ha lng ng huyt trong c th v cc bnh v ng tiu ha. 1.6 Gii thiu v qu trnh trch ly.

1.6.1 Bn cht. Bn cht ca qu trnh trch ly (qu trnh chit) l s rt cht ha tan trong cht lng hay cht rn bng mt cht ha tan khc (gi l dung mi) nh qu trnh khuch tn gia cc cht c nng khc nhau [4]. Trch ly cht ha tan trong cht lng c gi l trch ly lng, cn trch ly cht ha tan trong cht rn l trch ly cht rn. 1.6.2 Phm vi s dng ca qu trnh. Trong cng ngh thc phm nhm mc ch sau: Khai thc l mc ch ch yu. Ph bin l trch ly cc nguyn liu dng rn

nh: ht du, cc nguyn liu tinh du nh l, r, cy, hoa hoc qu; trch ly cc loi c nh c ci ng, trch ly ma. Cng vi mc ch ny c th phi hp vi qu trnh khc nng cao hiu sut thu sn phm. V d nh phi hp vi qu trnh p trong sn xut ma ng [4]. Trch ly cn nhm mc ch chun b cho cc qu trnh tip theo, v d nh

ngm cc loi ht (cc loi u, ng, thc,) trc khi ch bin; ngm cc loi c (sn, khoai,) lm yu cc lin kt trong vt liu, ng thi trch ly mt s phn t vo dung mi ngm (v d: ha tan HCN khi ngm c sn) [4].

31

Vi mc ch thu nhn sn phm nh tch penicillin t dung dch ln men

sn xut nc chm bng phng php m trch ly, hoc trch ly trong qu trnh sn xut c ph ha tan, trch ly khi ngm qu, ngm tm cc loi thuc b v thuc cha bnh, [4]. 1.6.3 Cc yu t nh hng n qu trnh chit. Thc cht qu trnh trch ly l qu trnh khuch tn. V vy s chnh lch nng gia hai pha (gradient nng ) chnh l ng lc ca qu trnh. Khi chnh lch nng ln, lng cht trch ly tng; thi gian trch ly gim ta thc hin bng cch tng t l dung mi so vi nguyn liu. Qu trnh trch ly polyphenol (phlorotannin) hin nay c thc hin bng cc loi dung mi. Nguyn tc ca trch ly bng dung mi da trn s thm thu dung mi vo t bo, cht cn trch ly ha tan vo dung mi v khuch tn ra ngoi t bo. Qu trnh trch ly bng dung mi c u im l thit b n gin, c th x l mt lng ln nguyn liu v c th s dng quy trnh lin tc. Cc yu t nh hng n qu trnh trch ly bng dung mi: La chn dung mi trch ly: Dung mi trch ly l dung mi ha tan c hp

cht cn trch ly. Cc polyphenol (phlorotannin) l cc hp cht phn cc nn ch yu s dng cc dung mi phn cc nh: nc, ethanol, acetone, ethyl acetate Din tch tip xc gia nguyn liu v dung mi. Cn tng din tch ny

lm tng hiu qu ca qu trnh. Vi cc nguyn liu rn cn tng din tch tip xc gia chng v dung mi. iu ny thc hin bng cch nghin nh, thi nh, bm nh vt liu. N cn lm ph v cu trc t bo, thc y qu trnh tip xc trit gia dung mi v vt liu. Tuy nhin kch thc v hnh dng ca vt liu khi lm nh cng c gii hn v nu chng qu mn s b lng ng ln lp nguyn liu, lm tc cc ng mao dn hoc b dng dung mi cun vo mixen (hn hp) lm cho dung dch c nhiu cn gy phc tp cho qu trnh x l tip theo [4]. Khuy trn: Khuy trn gip tng cng hiu qu trch ly, do tng cng

qu trnh truyn khi. pH: Polyphenol (phlorotannin) rt bn pH acid v km bn pH bazo. Khi

thc hin trch ly pH thp s trnh c hin tng oxy ha polyphenol v tng

32

hiu qu qu trnh trch ly. Tuy nhin, cng c nghin cu cho rng nh hng ca pH khng lm nh hng nhiu n hiu qu trch ly, ch cn m bo pH trung tnh hoc thp hn. Tnh cht ca vt liu cng nh hng ln n hiu sut trch ly. Vt liu l

hn hp lng- lng hoc hn hp rn lng cng vi mt dung mi hoc tp hp mt s loi dung mi. Chng c ha tan khc nhau, nng cc cht khc nhau v c tc dng tng h, khuch tn vo nhau. Nh khi trch ly u nu m nguyn liu gim, tc trch tng ln, v m tc dng vi protein v cc cht ho nc khc ngn cn s dch chuyn ca dung mi thm su vo trong nguyn liu, lm chm qu trnh khuch tn [4]. Nhit c tc dng tng tc khuch tn v gim nht, phn t cht

ha tan chuyn ng d dng khi khuch tn gia cc phn t dung mi. Tuy nhin nhit l mt yu t c gii hn. V khi nhit qu cao c th xy ra cc phn ng khc khng cn thit (lm oxi ha polyphenol cn trch ly), gy kh khn cho qu trnh cng ngh [4]. Do trong ton b qu trnh nhit khng nn vt qu 800C. S dng cht khng oxy ha h tr: Vic s dng cc cht khng oxy ha b

sung nh acid ascorbic, acid citric, c xc nhn lm tng hiu qu trch ly cng nh lm gim s oxy ha polyphenol. Yu t thi gian cng nh hng n qu trnh trch ly. Khi thi gian tng

ln, lng cht khuch tn tng, nhng thi gian phi c gii hn. V khi t c mc trch ly cao nht, nu ko di thi gian s khng mang li hiu qu kinh t. 1.6.4 Tng quan v dung mi chit. Dung mi: Dung mi l pha c nng thp. Dung mi l chit cc hp cht ra khi nguyn liu v thay i ty theo bn cht ca mi loi nguyn liu. C s la chn mt dung mi chit l tnh phn cc ca hp cht cha trong nguyn liu v ca dung mi. Dung mi phn cc v khng phn cc [6].

33

Qu trnh hnh thnh mt l ty thuc vo c tnh ca cht tan v dung mi. hnh thnh mt dung dch trc ht phi c s ph v cc dy ni lin kt trong hp cht tan v trong dung mi hnh thnh dy ni lin kt mi gia cht tan v dung mi. hiu v tnh phn cc ca dung mi, ta c th so snh gia 2 cht ete v nc: Hng s in mi ca nc 200C l 80.4 trong khi ca ete l 4.34. Nguyn t hydro ca phn t nc c kh nng lin kt vi mt nguyn t mang in m ca mt hp cht khc, hnh thnh dy ni lin kt hydro trong dung dch nc, trong khi ete etylic khng c s lin kt ny. Dy ni hydro hnh thnh nh hng n tnh ha tan ca hp cht i vi dung mi. Nc c tc dng va nh mt acid va nh mt bazo, cn ete ch l bazo rt yu v khng hot ng nh mt acid. Do nhng c tnh trn, nc c xem nh mt dung mi phn cc mnh cn ete l mt loi dung mi khng phn cc. Cc dung mi phn cc mnh ngoi nc ra cn c cc ancol bc thp nh: methanol, ethanol, propanol, Cc dung mi khng phn cc ngoi ete cn c cc hydrocacbua nh ete du, benzene, toluene, hexan,cc cht nm gia hai nhm ny gi l cc cht phn cc yu hoc va nh ethyl acetate, clorofoc, acetone, diethyl clorua. Khi la chn dung mi cn p ng c cc yu cu sau y [4]. Phi c tnh ha tan chn lc, tc l ha tan tt cc cht cn tch m

khng c ha tan hoc ha tan rt t cc cht khc. y l tnh cht rt c bn khng th thiu c khi la chn dung mi. Khng c tc dng ha hc vi cc cu t ca dung dch. Nu trch ly lng yu cu khi lng ring ca dung mi khc xa vi

khi lng ring ca dung dch. Tuy nhin cng c loi thit b trch ly dung dch c khi lng ring rt gn nhau. Khng ph hy thit b, dng c. Khng b bin i thnh phn khi bo qun.

34

chy.

Khng c khi thao tc, khng to hn hp n vi khng kh v kh

R tin, d kim. Dung mi phi c trch ly ra sau qu trnh ly bng phng php

un nng, chng ct hoc sy. Sau khi tch khng li mi v l v khng gy c cho sn phm. Trong cng ngh thc phm dung mi trch ly ph bin l nc (nh trch ly c ci ng, trch ly ma sau khi bm nh, trch ly c ph, ngm nc tinh ch cc loi tinh bt,). Vn quan trng cn gii quyt l cht lng nc thc phm. Bn cnh dung mi trch ly c th l dung dch ng trong qu trnh ngm cc loi qu, tm cc loi hng,; c khi dung mi l mt s dung dch ha cht c nng rt long nh: dung dch CuSO4 1%, dung dch nc vi long. Cc cht tan trong nc v dung mi phn cc [6].

Cc cht in ly nh cc mui v c u tan trong dung mi phn cc. Cht phn cc: Cc hp cht hu c ni chung khng ion ha nhng nu chng c cha cc nhm hoc nguyn t mang in m c th hnh thnh dy ni hydro vi phn t nc th chng s tan c trong nc. Nhng nhm c kh nng to dy ni hydro nh: -OH, CO, NO, NH2 v cc halogen gi l nhm phn cc. Cng c nhiu nhm phn cc th phn t y cng d ha tan trong nc. Nhng nu mch hydro cacbon ca phn t cng di th ha tan cng gim. Thc nghim cho thy, mt nhm phn cc trong phn t c kh nng hnh thnh lin kt hydro vi phn t H2O s lm cho phn t y tan c trong nc nu s cacbon ca mch khng qu 5 hoc 6 nu phn t c thm mch nhnh. Nu phn t c nhiu nhm phn cc (2 nhm tr ln) th t l ny gim xung. Mt nhm phn cc cho 3 hoc 4 cacbon trong mch th phn t y tan c trong nc. 1.6.5 Tm hiu dung mi chit trong ti. Nc. Phn t nc c cu to t mt phn t Oxy v hai phn t Hydro, m i electron trong mi trng lin kt ny b ko lch v pha Oxy nn phn t nc c tnh phn cc, do nc d lin kt vi cc nguyn t, phn t v cc cht khc. Mt khc cc phn t nc lin kt vi nhau bng cc lin kt Hydro (lin kt km

35

bn) nn cc phn t, cc nguyn t hoc cc cht khc d b gy lin kt to nn cht mi [19]. Nc s dng l nc c cung cp t nh my nc ca thnh ph. Nc t cc tiu chun cho php ca nc dng trong sinh hot v ch bin thc phm ca Vit Nam theo tiu chun ca B y t. Phng php chit xut. Chit xut (trch ly) l mt qu trnh tch chit hon ton cc cht c trong hn hp. Cc cht ny c ha tan khc nhau trong mt dung mi hoc trong nhng h dung mi khc nhau. Qu trnh chit xut l mt qu trnh phc tp, c th xy ra c l nh hin tng khuch tn, ha tan, thm thu, thm tch,ca cht tan vo dung mi. Phng php chit xut l bao gm c vic chn dung mi, dng c chit v cch chit. Mt phng php chit xut thch hp ch c th c hoch nh khi bit r thnh phn ca cc cht cn chit trong nguyn liu ra. Mi loi hp cht c ha tan khc nhau trong tng loi dung mi. V vy khng th c phng php chit xut chung p dng cho tt c cc nguyn liu. Phng php thc hin qu trnh trch ly: Cc cng on ch yu ca qu trnh trch ly l: Cho hn hp cn trch ly tip xc vi dung mi ri tch cc pha, sau tch v thu hi dung mi v tin hnh phn tch ring. Phng php ngm: Nguyn liu c ngm vi dung mi trong bn chit

nhit phng, thnh thong o trn. Thi gian ngm ty thuc vo loi nguyn liu. C th thc hin qu trnh trch ly gin on hoc lin tc; trch ly mt hoc

nhiu bc, dung mi c th chuyn ng cng chiu hoc ngc chiu. C th chit nhit thng hoc chit nng. Hai cch chit nhit

thng l ngm kit v ngm phn on. Phng php ngm kit cho kt qu tt hn v chit c nhiu hot cht v t tn dung mi, nht l khi p dng cch chit ngm kit ngc dng. Khi chit nng, nu dung mi l cc cht bay hi th p

36

dng cch chit lin tc hoc chit hi lu. Nu dung mi l nc th sc hoc hm phn on. Phng php ngm kit v phng php ngc dng: c thc hin trong

bn chit c van iu chnh dch chit chy xung theo mong mun sau khi ngm nguyn liu vi dung mi mt thi gian nht nh. Dch rt ra t bn th nht dng ngm nguyn liu bn th hai, tip tc thc hin cho n bn chit sau cng. Phng php ny c s dng rng ri trong sn xut.

37

CHNG 2: I TNG V PHNG PHP NGHIN CU

2.1 i tng nghin cu. i tng nghin cu l loi rong M (Sargassum serratum) mc t nhin vng bin Nha Trang Khnh Ha. Rong khi thu nhn c ra qua loi b cc cht bn c hc nc bin v lm kh n m bo qun.
Hnh 2.1: Rong M (Sargassum serratum)

2.2 Ha cht v dng c th nghim. Ha cht: Cc loi ha cht s dng cho ton b qu trnh thc hin cc th nghim u l ha cht tinh khit t tiu chun dng cho phn tch do Trung Quc sn xut. Dng c th nghim: S dng cc thit b hin c ca phng th nghim ha phn tch, trin khai cng ngh ti Vin nghin cu v ng dng cng ngh Nha Trang nh: cc thy tinh, bnh tam gic, a thy tinh, bnh nh mc, giy lc, bnh cu, cn, t sy, bp in, my so mu, thit b ly tm, my c quay, t lnh, 2.3 a) Phng php nghin cu S tin hnh nghin cu.
Kho st phng php o Phng php Folin Ciocalteu

La chn nguyn liu

Rong kh

Kho st qu trnh trch ly

Thu nhn phlorotannin

ng dng trong sn phm

38

b) lc 6). c)

Cc phng php phn tch ha hc v vt l. Xc nh hm lng m bng phng php sy (ph lc 1) Xc nh hm lng protein bng phng php Kjeldadl (ph lc 2). Xc nh hm lng lipid tng theo phng php Bligh v Dyer hn hp Xc nh hm lng Iod theo phng php chun (ph lc 4). Xc nh hm lng tro ton phn (ph lc 5). Xc nh hm lng x th theo Crudefibre v nh lng carbohydrate (ph Xc nh hm lng Fucoidan (ph lc 7). Xc nh hm lng Alginate (ph lc 8). Xc nh hm lng phlorotannin trong dch chit (ph lc 9). Phng php cm quan. Tin hnh nh gi cm quan sn phm ung theo phng php cho im

chloroform/methanol (ph lc 3).

cht lng theo TCVN 3215- 79. Phng php ny c s dng nh gi tng qut mc cht lng ca mt sn phm so vi tiu chun hoc so vi mt sn phm cng loi trn tt c cc ch tiu cm quan: mu sc, mi, v, trng thi. Tnh trng cht lng ca mi ch tiu c nh gi bng im. Gi tr im tng theo mc tng ca cht lng. Ty theo sn phm v quc gia m thang im s dng rt khc nhau (thang 10, 20, 50, thm ch l 100). Do cc ch tiu c vai tr i vi cht lng chung ca sn phm mc khc nhau nn cc gi tr cho c i vi mi ch tiu c nhn thm mt gi tr tng ng gi l h s trng lng. Cc ch tiu c vai tr ln hn th c h s trng lng cao hn. Vic xc nh h s trng lng cho mi ch tiu ca mi loi sn phm l rt kh. Cc h s ny thng c xc nh theo kinh nghim, phng php iu tra kt hp vi phng php chuyn gia trn c s thng k. Khi nh gi cht lng cm quan bng mt hi ng th im cht lng ca ch tiu no l im trung bnh ca mi thnh vin nhn vi h s trng lng ca n. Tng im ca cc ch tiu l im cht lng sn phm. im ny quyt nh mc cht lng ca sn phm c nh gi.

39

Vit Nam, phng php nh gi cm quan sn phm thc phm thc hin theo Tiu chun Vit Nam TCVN 3215 79. y l tiu chun s dng h im 20 xy dng trn mt thang thng nht 6 bc 5 im (t 0 n 5) trong im 0 ng vi cht lng sn phm b hng, cn t im 1 n im 5 ng vi mc khuyt tt gim dn. im 5 sn phm coi nh khng c sai li v khuyt tt no, trong tnh cht ang xt, sn phm c tnh tt c trng v r rt cho ch tiu . Tng h s trng lng ca tt c cc ch tiu c nh gi cho mt sn phm bng 4. Qu trnh nh gi phi c thc hin trong phng phn tch cm quan t yu cu. Vic chun b mu phi ph hp vi tng loi sn phm theo quy nh cht ch. Hi ng phi gm t 5 n 12 ngi chuyn gia c hiu bit v sn phm c nh gi. Hi ng c ch tch v th k lnh o hi ng trong qu trnh lm vic. Khi nh gi, cc thnh vin lm vic c lp, cho im vo phiu v np cho th k sau gi lm vic. Th k hi ng s tng kt im ca cc thnh vin v t tnh ra im cht lng ca sn phm. Theo h im 20, cht lng sn phm c chia ra 6 mc theo bng sau: Bng 2.1: Cc mc cht lng Mc Tt Kh Trung bnh Km Rt km Hng im 18.6 20.0 15.2 18.5 11.2 15.1 7.2 11.1 4.0 7.1 im trung bnh cha c trng lng Cc ch tiu quan trng nht 4.7 Cc ch tiu quan trng nht 3.8 Mi ch tiu 2.8 Mi ch tiu 1.8 Mi ch tiu 1.0

0.0 3.9 Sn phm t cht lng khi im trung bnh cha c trng lng ca mt

ch tiu bt k phi t nh nht l 2.8 v im cht lng khng nh hn 11.2. Nu hi ng thng nht nh cho mt ch tiu no 0 im th im chung bng 0 v sn phm coi nh hng. Nu thnh vin no cho im lch qu 1.5 im trung bnh cha c trng lng ca hi ng th im ca thnh vin b loi.

40

Bng 2.2: H s trng lng ca sn phm

Ch tiu im cht lng 5 Sn phm ng nht trong sut, khng c bt Trong sng, c trng cho sn phm ung rong bin. Mu nc vng sng ng, hp dn. 4 Sn phm trong, khng bt 3 Sn phm hi c 2 Sn phm c, c cn 1 Sn phm cn nhiu, nc c. 0 Sn phm qu nhiu cn, c vng, mc ni trn b mt. Sm en do qu nhiu cn bn. Mu ca sn phm h hng.

Trng thi

Mu sc

Trong sng, kh snh, c trng cho sn phm ung rong bin. Mu nc tng i.

Trong sng, c trng ca sn phm. Mu nc hi nht.

B c, khng c trng cho sn phm, c cn bn.

c ti, nhiu cn bn

Mi

Thm t Thm t nhin, hi nhin, gy n ha, hp tng kh hi dn, d ha v hp chu, c dn c trng trng cho cho sn sn phm, phm. khng c mi l hoc mi do khuyt tt. Mi thm mnh v bn. Ngt du, c trng cho sn phm, hi ha gia v v mi. Sn phm ngon, hp dn.

Mi thm tng i, c thong mi l.

Km thm, l mi l v mi do khuyt tt, khng c trng cho sn phm.

L r mi l v cc mi do khuyt tt.

Mi tanh kh chu ca sn phm hng.

Ngt du, c Ngt du, Ngt du, C v l V ca sn trng cho sn c cm gic nhng c hoc do cc phm hng. phm, kh xut hin v ln v khc. khuyt tt hi ha gia khc. khc gy v v mi. cm gic Sn phm kh chu. tng i hp dn.

41

d)

Phng php phn tch vi sinh Ch tiu vi sinh vt l ch tiu rt quan trng nh gi cht lng v v sinh

an ton thc phm. i vi sn phm nc ung rong bin ng chai: e) Xc nh tng s vi sinh vt hiu kh bng phng php TCVN 4884:2005. Xc nh Colifom v E. coli bng phng php TCVN 6848:2005. Xc nh tng s nm men, nm mc bng phng php TCVN 5166:1900. Xc nh S.aureus bng phng php TCVN 4830-1:2005. Xc nh Strepstococcuss faecalis bng phng php TCVN 6189-2:1996. Xc nh P.aeruginosa bng phng php 3347/2001/Q. Xc nh Cl.perfringens bng phng php TCVN 4991:2005. Phng php x l s liu. X l s liu theo phng php thng k. Mi th nghim u tin hnh 3 ln, v kt qu l trung bnh cng ca cc ln th nghim. S liu c x l v th trn phn mm MS. Excel. 2.4 2.4.1 Quy trnh trch ly v thu nhn phlorotannin. S quy trnh Rong kh

Ra sch Ct nh Dung mi Trch ly Lc C c Ly tm B

Cht chit Hnh 2.2: S quy trnh thu nhn phlorotannin t rong Sargassum Serratum.

42

2.4.2 Thuyt minh quy trnh Nguyn liu: Thu mua rong kh c x l v c ngun gc r rng. Mc ch ca vic la chn rong kh: hm lng m trong rong nh, gp phn quan trng trong vic c ch s pht trin ca vi sinh vt sng trn rong, c ch qu trnh sinh tng hp v chuyn ha trong rong. T rong c bo qun tt hn, m bo cho qu trnh thc hin khng gp gin on v vn nguyn liu. ng thi, khi hm lng m trong rong nguyn liu gim, tc trch ly tng ln v m tc dng vi protein v cc cht ho nc khc ngn cn s dch chuyn ca dung mi thm su vo trong nguyn liu, lm chm qu trnh khuch tn. Ra sch. Rong s c ra sch nhm loi b tp cht v mui bm dnh trn rong, to iu kin cho qu trnh chit v lc c thc hin d dng. Ct nh. Sau khi ra sch, rong c lm ro s b ri ct vi kch thc thun li cho qu trnh chit rt. Lm nh rong nhm mc ch tng din tch tip xc ca rong nguyn liu v dung mi, ng thi lm ph v cu trc t bo rong, to iu kin thun li cho cc cht ha tan trong dung mi. Tuy nhin, ta khng nn xay hoc nghin rong qu nh, bi kch thc bt rong qu nh, bt kn l ca giy lc lm cn tr qu trnh lc. Trch ly Phlorotannin. Phlorotannin l hp cht hu c c kh nng tan trong mt s loi dung mi phn cc. Nc l dung mi phn cc nn c s dng trch ly cc cht phn cc c trong rong, c th l phlorotannin. Cc yu t nh mi trng chit, nhit , t l nguyn liu: dung mi, thi gian chit u c nh hng n kh nng ha tan ca phlorotannin trong qu trnh chit. V vy ti cng on chit ny ta s tin hnh cc th nghim nghin cu nh hng ca mt s yu t n kh nng chit phlorotannin. Lc Sau khi chit, tin hnh lc nhm thu hi nhng cht ha tan c trong dung mi. ng thi loi b c nhng tp cht khng tan khc (cn, b rong).

43

C c Sau khi lc, dch rong c em i c c, nhm mc ch loi b bt dung

mi trong dch lc nh qu trnh bay hi dung mi bi nhit v p sut, to iu kin thun li cho cng on sau. Qu trnh c c ph thuc vo nhit , thi gian, ch khuy o, p sut ca thit b,Trong qu trnh c c, dung mi trong dch rong bay hi nn mu sm li, th tch dch gim dn. Nu c c nhit v thi gian khng thch hp s gy hao ht hoc lm bin tnh cht tan cn chit. V vy khng nn c c nhit qu cao v thi gian di. Sau khi c c, dch chit c em i phn tch thnh phn ri qua cng on ly tm. Ly tm Sau qu trnh c c, tin hnh ly tm dch rong nhm loi b ht cc tp cht hoc b rong cn st li sau khi lc v c c. Bn cht ca qu trnh ly tm l tch d dng cc phn t khc nhau v kch thc v trng lng (khi lng ring) trong dch chit, di tc dng ca lc ly tm v trng lc ca Tri t (lc ly tm ln hn trng lc rt nhiu). 2.5 Cc yu t nh hng n qu trnh trch ly Phlorotannin. Kho st qu trnh trch ly Trch ly nhm mc ch tch phlorotannin ra khi rong bng cch s dng dung mi ha tan da trn c s chnh lch nng . Trong qu trnh chit, vic la chn ra loi dung mi chit thch hp nht ha tan tt cht tan cn xc nh l mt yu t rt quan trng. Qua bn cht ca mt s loi dung mi v ca cht tan, ta thy dung mi c th ha tan tt phlorotannin l cc dung mi c tnh phn cc, v vy ti chn dung mi l nc.. Phng php kho st: khi kho st mt yu t no (nhit , thi

gian) th cc yu t cn li c nh. Yu t no kho st xong th s c c nh gi tr ti u tip tc kho st n cc yu t khc. 2.5.1 Tin hnh kho st iu kin mi trng (pH) trch ly. Khi lng mu cho mi th nghim l 10(g). Tin hnh ngm rong trong dung mi l nc.

44

Chun b 11 mu th nghim theo cc pH nh sau: 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0. Khi tin hnh th nghim ta c nh cc thng s: T l nguyn liu: dung mi l 1:20 Nhit trch ly l 700C Thi gian trch ly l 120 pht. Dch chit thu c em i xc nh hm lng phlorotannin v carbohydrat. T la chn pH chit thch hp. Trch ly

1.0

1.5

2.0

2.5

3.0

3.5

4.0

5.0

6.0

7.0

8.0

o OD

Chn pH ti u Hnh 2.3: S b tr th nghim xc nh nh hng ca pH n kh nng trch ly (chit). 2.5.2 Tin hnh kho st t l dung mi trch ly. Vic la chn t l nguyn liu: dung mi (NL/DM) c nh hng ln ti qu trnh chit. Nu lng dung mi qu t qu trnh chit s khng trit , lng cht ha tan bn trong nguyn liu cn nhiu (do chnh lch nng cht tan nh). Ngc li nu lng dung mi qu ln, kh nng chit trit hn, lng cht tan ha tan nhanh hn, tuy nhin li nh hng n chi ph dung mi v ko di thi gian thc hin cng on c c. V vy cn la chn t l dung mi sao cho thch hp nht.

45

Chun b 10 mu th nghim theo cc t l khi lng nguyn liu: dung mi nh sau: 1: 5, 1: 10, 1: 15, 1: 20, 1: 25, 1: 30, 1: 40, 1: 50. C nh cc thng s: pH c xc nh th nghim 2.5.1 Nhit trch ly: 700C. Thi gian trch ly: 120 pht. Dch chit thu c em xc nh hm lng phlorotannin v carbohydrate. T la chn t l nguyn liu: dung mi thch hp.

Trch ly

1: 5

1: 10

1: 15

1: 20

1: 25

1: 30

1: 40

1: 50

o OD

Chn t l dung mi ph hp

Hnh 2.4: s b tr th nghim xc nh nh hng ca t l dung mi n kh nng trch ly. 2.5.3 Tin hnh kho st nhit trch ly. Nhit c nh hng n kh nng ha tan cc cht trong rong nguyn liu vo mi trng chit. Khi nhit tng cc phn t khuch tn cng mnh, nht gim, lng cht tan vo mi trng chit cng nhiu. Tuy nhin nu nhit qu cao c th lm bin tnh mt s hp cht c hot tnh sinh hc hoc lm hao ht mt lng cht tan d bay hi khc. Nu nhit qu thp, lng cht tan khuch tn ra mi trng b hn ch. V vy cn xc nh nhit chit thch hp.

46

Chun b 7 mu th nghim theo cc nhit nh sau: 400C, 500C, 600C, 700C, 800C, 900C, 1000C. C nh cc thng s: PH c xc nh th nghim 2.5.1 T l nguyn liu: dung mi c xc nh th nghim 2.5.2 Thi gian trch ly: 120 pht. Dch chit thu c em xc nh hm lng phlorotannin v carbohydrate. T la chn nhit trch ly thch hp.

Trch ly

400C

500C

600C

700C

800C

900C

1000C

o OD

Chn nhit ti u Hnh 2.5: S b tr th nghim xc nh nh hng ca nhit n kh nng chit. 2.5.4 Tin hnh kho st thi gian trch ly. Th nghim nhm xc nh thi gian chit ti u thu c hm lng phlorotannin cao nht. Thi gian chit phi hp l, nu ko di thi gian chit lng cht ha tan dung mi ht, ng thi s nh hng n thi gian thc hin v chi ph cho ton b qu trnh. Chun b 12 mu th nghim theo cc thi gian nh sau: 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240 (pht).

47

C nh cc thng s: pH c xc nh th nghim 2.5.1 T l nguyn liu: dung mi c xc nh th nghim 2.5.2 Nhit trch ly c xc nh th nghim 2.5.3 Dch chit thu c em xc nh hm lng phlorotannin v carbohydrate. Trch ly

20

40

60

80

100

120

140

160

180

200

220

240

o OD

Chn thi gian ti u Hnh 2.6: S b tr th nghim xc nh nh hng ca thi gian n kh nng chit. 2.5.5 Tin hnh xc nh hiu sut trch ly Tin hnh trch ly mu iu kin ti u kho st trn. Sau khi trch ly 1 ln xong th tin hnh lc thu ly dch lc. Phn b tip tc c s dng trch ly li (ln 2, 3, 4) cc iu kin tng t nh trch ly ln 1. Vic trch ly c lp li 4 ln trn cng mt mu. o OD ca dung dch sau mi ln trch ly. T so snh vi hm lng ban u tnh c hiu sut trch ly. C nh cc thng s: pH c xc nh th nghim 2.5.1 T l nguyn liu: dung mi c xc nh th nghim 2.5.2 Nhit trch ly c xc nh th nghim 2.5.3

48

Thi gian trch ly c xc nh th nghim 2.5.4

Dch chit thu c em xc nh hm lng phlorotannin v carbohydrate. Trch ly

1 ln

2 ln

3 ln

4 ln

o OD

Chn s ln trch ly Hnh 2.7: s b tr th nghim xc nh nh hng ca s ln chit 2.5.6 Th nghim xc nh nhit c c. Qu trnh c c l tch bt lng dung mi c trong dch chit to iu kin thun li cho cc cng on sau. Ta cn xc nh nhit c c thch hp nhm hn ch nh hng ca yu t nhit n cht tan cn chit. B tr th nghim: C c

Nhit c c (0C)

40

50

60

70

Xc nh hm lng phlorotannin, Carbohydrate

La chn nhit c c thch hp Hnh 2.8: S b tr th nghim xc nh nhit c c.

49

Cch tin hnh: Sau khi dch rong c lc, ta chia dch rong thnh 4 th tch bng nhau (khong 500 ml) ri tin hnh c c ti cc nhit 400C, 500C, 600C v 700C bng thit b c quay (iu kin chn khng ti nhit 400C). Lm th nghim ny khi la chn c cc yu t nh hng n qu trnh chit nh: loi dung mi, t l nguyn liu: dung mi, nht chit v thi gian chit. 2.5.7 Th nghim xc nh thi gian ly tm Qu trnh ly tm l tch ht cn (b rong) cn st li trong dch chit v cc cn mui hnh thnh trong qu trnh c c . Dch chit thu c s trong hn, sng mu hn. Ta cn xc nh thi gian ly tm thch hp tch trit cn trong dch rong, ng thi tit kim c thi gian v chi ph thc hin qu trnh. B tr th nghim: Thi gian ly tm (pht)

10

15

20

25

Kim tra trong ca dch rong

La chn th nghim thi gian ly tm Hnh 2.9: S b tr th nghim xc nh thi gian ly tm Cch tin hnh: Dch rong sau khi c c tch bt dung mi c chia thnh 5 mu v tin hnh ly tm tc 5000 vng/ pht ti cc mc thi gian: 5 pht,

50

10 pht, 15 pht, 20 pht, 25 pht. Sau kim tra trong bng cm quan v la chn thi gian ly tm thch hp. 2.6 ng dng cao chit phlorotannin vo qu trnh to sn phm: nc

ung rong bin ng chai. S quy trnh

Nguyn liu

Trch ly

Cao hlorotannin

Lc

Phi ch

Dch ng

Gia nhit, bi kh

Natribenzoat

Rt chai, y np

Chai, np

Thanh trng, lm ngui

Bo n

Thnh phm

Hnh 2.10: S quy trnh to sn phm nc ung rong bin ng chai.

51

Thuyt minh quy trnh

Nguyn liu: S dng nguyn liu rong kh sch, khng ln tp cht v cc

loi khc, khng b h hng, m mc,loi rong s dng c cha hm lng Phlorotanin cao v mt s thnh phn khong cht c li cho sc khe. Trch ly: Tch chit cc thnh phn c trong rong nguyn liu. Lc: Nhm loi b phn b v cc cu t l lng gy c, lm trong dch

trch ly. Phi ch: Dch trch ly c phi trn cng vi cao phlorotannin, dung dch

ng. Gia nhit, bi kh: To iu kin thun li cho qu trnh vo chai, do ui ht

khng kh ra khi chai. giai on ny c b sung Natribenzoate nhm mc ch bo qun sn phm. Rt chai, y np: s dng bao b thy tinh th tch 300ml vi chiu cao l

20 cm. Np chai lm bng st, c m km v c sn lp vecni chng r, m bo phi to kn cho chai thy tinh. Thanh trng, lm ngui: Tiu dit cc vi sinh vt gy h hng sn phm, ko

di thi gian bo qun. Bo n: Pht hin cc sn phm khng t cht lng trc khi a ra th

trng.

52

CHNG 3: KT QU NGHIN CU V THO LUN

3.1 Kt qu xc nh mt s thnh phn ca rong nguyn liu. Trong n ny ti la chn rong Nu Sargassum Serratum lm nguyn liu chit phlorotannin. V y l loi rong c tr lng ln v phn b rng ri ti vng bin Vit Nam. nh gi s b cht lng ca rong Sargassum Serratum, ti tin hnh xc nh mt s thnh phn ha hc ca nguyn liu ny v kt qu phn tch c ch ra trn bng 3.1 nh sau: Bng 3.1 Thnh phn ha hc chnh ca rong nguyn liu STT 1 2 3 4 5 6 7 8 9 Phlorotannin m Tro Protit th Lipid It Alginate Laminaran Fucoidan Thnh phn Hm lng (% cht kh) 0.8 11.5 35.1 4.7 0.7 0.1 25.9 0.1 1.7

Nhn xt: T kt qu trong bng 3.1 ta thy, thnh phn ha hc chnh ca rong Sargassum Serratum bao gm cc cht c hm lng ln nh alginate, khong v cc thnh phn khc chim t 0.1% n 11.5% nh protein, fucoidan, phlorotannin, cc cht tan trong cn nh lipid, iot. Nhng thnh phn ny c th tan trong cc loi dung mi phn cc, khng phn cc. Nn khi chit phlorotannin s xy ra ng thi qu trnh chit cc cht trn, dch chit thu c bao gm phlorotannin v cc cht i km, s c mt ca nhng cht ny trong dch chit chim mt hm lng ng k. V vy tng hm lng phlorotannin chit v gim cc cht i km, ta tin hnh la chn mt s thng s thch hp c th chit c hm lng phlorotannin l nhiu nht.

53

3.2

Kt qu kh mi ca rong bin. Trong n ny ti s dng ngay dch chit c c phi ch to nc

ung, do , sn phm ung s c mi tanh ca rong bin do s c mt cc dn xut ca pyridine nh 2,4-di-isopropenylpyridine. y ti s dng tc nhn kh mi l cht H2O2 v tin hnh th nghim nh sau: %. Kt qu thu c nh sau: Bng 3.2 nh hng ca nng dung dch H2O2 n kh nng kh mi Dung dch H2O2 Cm quan (mi) 0,1 % Mi tanh 0,3 % Mi tanh 0,5 % Mi tanh nh 0,7 % 0,9 % 1,2 % Nhit kh mi Thi gian kh mi : Nhit phng :10 pht

Thay i nng dung dch H2O2 :0,1%; 0,3%; 0,5%; 0,7%; 0,9 % v 1,2

Mi thm c trng ca rong bin, khng c v tanh

Da trn s liu dn trn bng 3.2, ti chn iu kin kh mi l ngm rong trong dung dch H2O2 trong 10 pht ti nhit phng, sau ra sch bng nc v tin hnh chit. 3.3 Kt qu xc nh thng s thch hp ca qu trnh chit phlorotannin. Theo mt s tc gi [1], [2], [6] th phlorotannin tan trong dung mi phn cc v vi rong M sinh trng ti Vit Nam th theo kt qu ca [7] dung mi chit phlorotannin thch hp l axeton:nc vi t l 7:3. Tuy nhin khi s dng dung mi ny s ch chit c phloroatannin m khng chit c mt s cht c hot tnh trong rong nh fucoidan, It, khongHn th na s dung dung mi axeton th hiu qu kinh t khng cao do gi thnh dung mi chit cao. Mc ch ca lun vn l thu nhn c thc phm ung c tc dng tt tng cng bo v sc khe. V vy ti la chn dung mi nc c th chit c phlorotannin v cc cht c hot tnh nh l fucoidan, It..Chnh v l do nu trn nn ti la chn dung mi l nc. Nc l dung mi c tnh phn cc mnh nn ngoi kh nng ha tan phlorotannin, n cn c th ha tan c nhiu thnh phn ha hc khc trong rong

54

nguyn liu. c bit nhng thnh phn tan c trong nc li chim mt lng ln trong rong nh: khong, acid fucxinic, fucoidan, laminarin, iod, sc t,[6]. Chnh v vy lng cht chit trong mi trng nc l ln. Mt khc nc li l dung mi d kim, r tin v an ton. ng thi cc thnh phn ny (nht l iod) to nn mu vng ca dch chit. Khi s dng dung mi nc, cc cht chit i km cng vi phlorotannin ch yu l fucoidan, laminaran v cht khong. V vy xc nh kh nng chit ti xc nh 02 thnh phn chnh l phlorotannin v fucoidan thng qua vic xc nh hm lng carbohydrate. tm iu kin chit thch hp ti tin hnh cc nghin cu sau: nh hng ca mi trng (pH) n kh nng chit. nh hng ca t l NL/DM (khi lng/th tch). nh hng ca nhit n kh nng chit. nh hng ca thi gian n kh nng chit. nh s ln chit (Hiu sut chit ly). Vi dung mi chit l nc, ta tin hnh nghin cu nh hng ca pH n kh nng chit. Kt qu c th hin hnh 3.1a; 3.1b v bng 3.3 (ph lc 11), bng 3.4 (ph lc 12).
Hm lng phlorotannin (mg/ml)
30 25 20 15 10 5 0 1 1.5 2 2.5 3 3.5 4 5 6 7 8

3.3.1 Kt qu xc nh nh hng ca mi trng (pH) n kh nng chit.

pH

Hnh 3.1a: Biu biu din nh hng ca pH trch ly n hm lng phlorotannin trong qu trnh chit.

55

Hm lng carbohydrate (mg/ml)

80 70 60 50 40 30 20 10 0 1 1.5 2 2.5 3 3.5 4 5 6 7 8

pH

Hnh 3.1b: Biu biu din nh hng ca pH trch ly n hm lng carbohydrate trong qu trnh chit. Nhn xt: Dng acid H2SO4 5% v NaOH 5% iu chnh pH ca dung dch v cc khong 1.0; 1.5; 2.0; 2.5; 3.0; 4.0; 5.0; 6.0; 7.0; 8.0. T kt qu nghin cu (ph lc 11, ph lc 12) v nhn vo th (hnh 3.1a, hnh 3.1b) ta thy khi gim pH t 8.0 xung 1.0 th hm lng phlorotannin v hm lng carbohydrate u tng , iu ny chng t trong mi trng acid th kh nng thu nhn hai thnh phn trn tng. iu ny l do phlorotannin tn ti trong cy rong di dng to lin kt cng ha tr vi fucoidan v alginate trong mng t bo, v vy khi gim pH ca dung mi chit s ph hy mng t ba v ph v lin kt cng ha tr ca phlorotannin dn n kh nng kh chit tng ln v hm lng cc cht carbohydrate v phlorotannin tng ln. Tuy nhin khi iu chnh pH xung qu su s lm cho t bo ca rong b co li mt nc dn ti cn tr qu trnh trch ly phlorotannin v carbohydrate vo dung mi. Do ti chn pH = 2 l pH ti u cho qu trnh chit. 3.3.2 Kt qu xc nh nh hng ca t l NL/DM (khi lng/th tch) n kh nng chit.

56

Dung mi chit l nc. Sau khi chn c pH chit = 2, ta tin hnh nghin cu nh hng ca t l nguyn liu/dung mi n kh nng chit. Kt qu c th hin hnh 3.2a; 3.2b v bng 3.5 (ph lc 13); bng 3.6 (ph lc 14).

Hm lng phlorotannin (mg/ml)

30 25 20 15 10 5 0 1:05 1:10 1:15 1:20 1:25 1:30 1:40 1:50

T l NL/DM
Hnh 3.2a: Biu biu din nh hng ca t l NL/DM n hm lng phlorotannin trong qu trnh chit.
100 90 80 70 60 50 40 30 20 10 0 1:05 1:10 1:15 1:20 1:25 1:30 1:40 1:50

Hm lng carbohydrate (mg/ml)

T l NL/DM
Hnh 3.2b: Biu biu din nh hng ca t l NL/DM n hm lng carbohydrate trong qu trnh chit.

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Nhn xt: T kt qu nghin cu (ph lc 13, ph lc 14) v th (hnh 3.2a, hnh 3.2b) cho thy: khi tng lng dung mi s dng th hiu qu trch ly phlorotannin tng th hin r nht khi tng t l t 1: 5 ln 1: 10; 1: 15 ln 1: 20; 1: 30 ln 1: 40; 1: 40 ln 1: 50; v tip tc tng t t l 1: 10 ln 1: 15 v 1: 20 ln 1: 30. Hm lng carbohydrate tng th hin r nht khi tng t l t 1: 5 cho n 1: 40; gim xung t l 1: 50. C th gii thch iu ny nh sau: Trong qu trnh trch ly bng dung mi th nhn thy rng khi lng dung

mi cng ln th hiu qu trch ly cng tng do chng lun to ra c mt s chnh lch nng cn thit bn trong v bn ngoi mi trng tc l lun c ng lc cho qu trnh. Khi t l NL/DM gim tc lng dung mi s dng tng, phlorotannin,

carbohydrate v cc cht tan c trong rong Sargassum Serratum s c iu kin ha tan tt vo dung mi bi lng dung mi ln s lm tng kh nng tip xc ca dung mi v cht tan, to ra c mt s chnh lch nng cn thit bn trong v bn ngoi mi trng tc l lun c ng lc cho qu trnh, t lm tng chnh lch p sut thm thu v s khuch tn ca cc cht tan c tron g t bo rong ra dung mi chit, lm cho hm lng cht tan, phlorotannin v carbohydrat c trong dch chit tng ln. Lc u nng cht tan, phlorotannin v carbohydrate c trong rong nhiu,

s khuch tn chng ra khi t bo cng nhanh v mnh. Nhng dn dn khi lng dung mi ngm kit vo trong cc thnh phn ca rong (s thm thu gn t trng thi bo ha) v nng cht tan cng nh phlorotannin v carbohydrate gim so vi ban u, th s khuch tn cc cht ny ra khi t bo cy rong cn g gim v chm hn. Do kh nng ha tan ca chng vo mi trng chit s gim v dn n n nh (lc ny chnh lch nng cht tan trong t bo rong v trong dch chit l rt nh). Tuy nhin nu chn lng dung mi qu ln (t l NL/DM= 1/30 hay 1/40) s gy lng ph dung mi v nng lng, ng thi lm ko di thi gian c c. Do vy, ti chn t l NL/DM l 1/20 lm t l chit thch hp cho qu trnh chit v l thng s c nh cho cc cng on tip theo.

58

3.3.3 Kt qu xc nh nh hng ca nhit n kh nng chit. Dung mi chit l nc. Sau khi chn c pH chit = 2 , t l NL/DM = 1/20 thch hp, ta tin hnh nghin cu nh hng ca nhit n kh nng chit trong nhng iu kin ny. Kt qu c th hin trn hnh 3.3a; 3.3b v bng 3.7 (ph lc 15); bng 3.8 (ph lc 16)
Hm lng phlorotannin (mg/ml)
30 25 20 15 10 5 0 40 50 60 70 80 90 100

Nhit trch ly (0C)

Hnh 3.3a: Biu biu din nh hng ca nhit chit n hm lng phlorotannin trong qu trnh chit.
Hm lng carbohydrate (mg/ml
80 70 60 50 40 30 20 10 0 40 50 60 70 80 90 100

Nhit trch ly (0C)

Hnh 3.3b: Biu biu din nh hng ca nhit chit n hm lng carbohydrate trong qu trnh chit. Nhn xt: T kt qu nghin cu (ph lc 15, ph lc 16) v th (hnh 3.3a, hnh 3.3b), ta thy hm lng phlorotannin trong dch chit tng ln khi ta

59

tng nhit t 400C n 700C v gim xung khi tng nhit t 700C n 1000C. Cn hm lng carbohydrate trong dch chit nm trong khong 51.35625 (mg/ml) n 72.1875 (mg/ml) (tng ln khi tng nhit t 400C n 500C ri gim khi tng ln 600C, sau li tip tc tng ln khi tng nhit ln 900C v gim xung khi tng nhit ln 1000C). C th gii thch iu ny nh sau: Khi tng nhit chit s lm gim nht ca dch, lm tng tc phn

ng gia cc thnh phn ha hc [4] trong rong vi nhau cng nh cc thnh phn vi hn hp dung mi chit. V tng hiu qu ca qu trnh trch ly. Tuy nhin nhit qu cao lm cho hin tng chuyn pha ca dung mi din ra mnh lit ng thi qu trnh oxy ha phlorotannin tng cao nn lm gim hiu qu trch ly. Khi tng nhit chit lm cho chuyn ng nhit tng, tc khuch tn

ca cc cht tan cng nh phlorotannin v carbohydrate t bn trong t bo rong ra mi trng chit s tng, lm tng s thm thu gia dung mi v t bo nguyn liu [4]. V th, hm lng phlorotannin v carbohydrate trong dch chit tng ln. Tuy nhin, nhit l mt yu t c gii hn. V nhit qu cao, chng ta phi cung cp thm nng lng, nh hng n gi thnh sn phm v c th xy ra cc phn ng khng mong mun, lm bin tnh hay thay i mt s thnh phn cht tan cn chit. Nhit qu cao lm cho hin tng chuyn pha ca dung mi din ra mnh lit, ng thi qu trnh oxy ha phlorotannin tng cao nn lm gim hiu qu trch ly. Cn nu nhit qu thp th li ko di thi gian trch ly nh hng n c qu trnh. Nhn chung c th thy l yu t nhit t nh hng n qu trnh trch ly. V vy, ti chn nhit chit 600C l nhit chit thch hp cho qu trnh chit v l thng s c nh cho cc cng on tip theo. 3.3.4 Kt qu xc nh nh hng ca thi gian n kh nng chit. Dung mi chit l nc. Sau khi chn c pH chit = 2 , t l NL/DM = 1/20 , nhit chit l 600C, ta tin hnh nghin cu nh hng ca thi gian n kh nng chit trong nhng iu kin ny. Kt qu c th hin trn hnh 3.4a; 3.4b v bng 3.9 (ph lc 17); bng 3.10 (ph lc 18).

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Hm lng phlorotannin (mg/ml

30 25 20 15 10 5 0 20 40 60 80 100 120 140 160 180 200 220 240

Thi gian trch ly (pht)

Hnh 3.4a: Biu biu din nh hng ca thi gian chit n hm lng phlorotannin trong qu trnh chit.
Hm lng carbohydrate (mg/ml)
90 80 70 60 50 40 30 20 10 0 20 40 60 80 100 120 140 160 180 200 220 240

Thi gian trch ly (pht)

Hnh 3.4b: Biu biu din nh hng ca thi gian chit n hm lng carbohydrate trong qu trnh chit. Nhn xt: T kt qu nghin cu (ph lc17, ph lc 18) v th (hnh 3.4a, hnh 3.4b), ta thy c rng khi thay i thi gian t 20 pht n 200 pht th tng hm lng phlorotannin v carbohydrate trong dch trch ly tng dn; t 220 pht n 240 pht th tng hm lng phlorotannin v carbohydrate trong dch trch ly gim xung. C th gii thch iu ny nh sau:

61

Khi thi gian chit ko di, gip dung mi thm thu vo trong tng t bo

rong qua cc mao qun, to iu kin thun li cho s khuch tn cht tan , phlorotannin v carbohydrate ra khi rong vo mi trng chit. Lc u lng cht tan, phlorotannin v carbohydrate c trong rong nhiu

nn kh nng ha tan ca n trong mi trng chit s ln. Nhng lng phlorotannin cng nh lng carbohydrate trong rong ch c mc gii hn nht nh. V vy hm lng ca chng ch tng mnh thi gian u v gim dn v sau, n mt thi im no khi s khuch tn cc cht ny xy ra hn ch th hm lng phlorotannin cng carbohydrate s khng thay i. Ta thy thi gian trch ly di th hm lng phlorotannin v carbohydrate

trong dch chit tng, nhng vic ko di thi gian li bt li v mt nng lng v nh hng ti thi gian thc hin quy trnh. Qua qu trnh thc nghim v do rong nguyn liu mi c thu v, cht lng cn tt nn ti chn thi gian chit ngn, 100 pht l thi gian chit thch hp cho qu trnh chit. 3.3.5 Xc nh hiu sut trch ly. Sau 4 ln trch ly th hm lng phlorotannin v carbohydrate trong nguyn liu cn li khng ng k, kt qu c th hin trn hnh 3.5a, 3.5b v bng 3.11 (ph lc 19); bng 3.12 (ph lc 20), do :
Hm lng phlorotannin (mg/ml
30 25 20 15 10 5 0 Ln 1 Ln 2 Ln 3 Ln 4

Ln trch ly

Hnh 3.5a: th biu din hm lng phlorotannin trong cc ln trch ly

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Hm lng carbohydrate (mg/ml)

70 60 50 40 30 20 10 0 Ln 1 Ln 2 Ln 3 Ln 4

Ln trch ly
Hnh 3.5b: th biu din hm lng carbohydrate trong cc ln trch ly. Tng lng phlorotannin trch ly c trong rong l 33.62 (mg/ml). Hiu sut trch ly phlorotannin ln 1 = 25.36 / 33.62= 75.43 (%) Hiu sut trch ly phlorotannin ln 2 = 4.20 / 33.62= 12.49 (%) Hiu sut trch ly phlorotannin ln 3 = 2.45 / 33.62 = 7.29 (%) Hiu sut trch ly phlorotannin ln 4 = 1.61/ 33.62 = 4.79 (%)
Hiu sut trch ly phlorotannin
Hm lng phlorotannin (mg/ml)
100 80 60 40 20 0 1 ln 2 ln 3 ln 75.43 87.92 95.21

Ln trch ly

Hnh 3.6a: th biu din hiu sut trch ly phlorotannin trong cc ln trch ly Tng lng carbohydrate trch ly c trong rong l 101.85 (mg/ml) Hiu sut trch ly carbohydrate ln 1 = 62.700/ 101.85 = 61.56 (%)

63

Hiu sut trch ly carbohydrate ln 2 = 20.0475/ 101.85 = 19.68 (%) Hiu sut trch ly carbohydrate ln 3 = 12.870 / 101.85 = 12.64 (%) Hiu sut trch ly carbohydrate ln 4 = 6.22875/ 101.85= 6.12(%)
Hiu sut trch ly carbohydrate
Hm lng carbohydrate (mg/ml
100 90 80 70 60 50 40 30 20 10 0 93.88 81.24 61.56

1 ln

2 ln

3 ln

Ln trch ly

Hnh 3.6b: th biu din hiu sut trch ly carbohydrate trong cc ln trch ly. Nhn xt: Nu trch ly 1 ln th phlorotannin v carbohydrate cn li trong b rt nhiu (8.26 (mg/ml) phlorotannin v 39.15 (mg/ml) carbohydrate). Nu trch ly 2 ln th phlorotannin v carbohydrate cn li trong b khng nhiu (4.06 (mg/ml) phlorotannin v 19.1025 (mg/ml) carbohydrate). Nu trch ly 3 ln th phlorotannin v carbohydrate cn li trong b rt thp (1.61 (mg/ml) phlorotannin v 6.2325 (mg/ml) carbohydrate). Nh vy sau 2 ln trch ly, lng phlorotannin v carbohydrate thu c t rong tng ng l 29.56 (mg/ml) v 82.7475 (mg/ml) (trong 10 (g) nguyn liu ban u). Do vy, chng ta nn tin hnh trch ly 2 ln thu hm lng phlorotannin v Carbohydrate c trong rong. 3.3.6 Kt qu xc nh thng s thch hp ca qu trnh c c. V ti chn t l nguyn liu: dung mi l 1:20 nn th tch dch chit ln v nng cht chit nh. V vy cn thit phi tng nng cht chit bng phng

64

php khc nhau. Trong lun vn ny, ti chn phng php c c. y l phng php s dng ph bin i vi cc cht c hot tnh sinh hc. xc nh nhit c c, mi mt mu ti ly 500 ml dch chit em c ti cc nhit 500C, 600C, 700C ti p sut thng v c ti nhit 400C di p sut 40 mmHg cho n khi th tch dung dch cn 1/10 th tch ban u (50 ml) v em cc mu i phn tch v nh gi cm quan. Kt qu th hin trn bng 3.13. Bng 3.13: nh hng ca nhit n hm lng Phlorotannin v Carbohydrate trong qu trnh c c. Nhit c c Stt Ch tiu phn tch Hm lng phlorotannin (mg/ml) Hm lng 2 carbohydrate (g/ml) 56910.25 56525.75 55425.25 55205.25 400C 500C 600C 700C

19.45

19.15

18.60

18.04

Cm quan Mu sc 3 Trng thi Nu Dng lng c cn mu trng Nu Dng lng cn mu trng Nu en Dng lng cn mu trng Nu en Dng lng cn mu vng

Hot tnh oxy ha tng s (mg axit ascorbic/g cht kh)

38,7 mg /g cht kh

38,1 mg /g cht kh

30,3 mg /g cht kh

18,2 mg /g cht kh

S liu bng 3.13 cho thy rng, hm lng carbohydrate v hm lng phlorotannin t thay i theo nhit chit. Tuy nhin cc ch tiu cm quan thay i r rt t mu nu sang nu en khi nhit c t tng t 400C n 70 0C.

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Nguyn nhn l do qu trnh c c l qu trnh lm bay hi cht lng khi dch chit nh s chnh lch nhit v p sut [4]. Tc chuyn ng nhit ca cc phn t cht lng nm gn mt thong mt thi im no gii hn nn cc phn t thot ra khi b mt tr thnh trng thi t do (hi). Khng nhng phn t trn mt thong bay hi m ch yu l cc bt hi to thnh trong cht lng. Chng l nhng tm to thnh hi nh lc y Acsimet c y ln mt thong v chng chuyn ng lin tc t bn trong lp cht lng ln b mt thong. iu kin to thnh bt hi l p sut bt bng p sut cht lng [4]. S bay hi dung mi lm gim th tch ca hn hp dch (gm cht tan v dung mi), t tng nng dung dch v lm cho mu dch sau khi c s sm li. Trong qu trnh c c, nhit si ca dch ph thuc vo p sut hi trn b mt, nng cht kh v tnh cht vt l, ha hc ca hn hp. Khi p sut hi trn b mt ca sn phm cng thp th nhit si ca dch s gim. Hay ni cch khc chng ta iu chnh nhit si bng cch thay i chn khng [6]. ng thi, khi nhit si thp tnh cht ca sn phm t b bin i v mu sc cng t bin i. Nhit si thp cn lm gim tc n mn ca vt liu ln thit b, ko di thi gian bn ca thit b [6]. Do vy ti la chn phng php c chn khng d dng iu chnh c nhit si bng cch gim p sut m khng cn tng nhit , hn ch c qu trnh oxy ha cht tan cng nh nh hng xu ca nhit cao n dch c. Bn cnh , tit kim thi gian v gim c chi ph nng lng trong qu trnh thc hin, ti tin hnh c dch rong nhit 500C trong thi gian khong 30 50 pht. Phn tch sn phm c c cho kt qu sau: Bng 3.14: Hm lng cc thnh phn (%) sau khi c c. Cc thnh phn Cht kh Phlorotannin Carbohydrate Protein Iod Hm lng (tnh theo %) 30 9.0 13.5 2.1

0.8 Nh vy, s dng keo rong ny, ti to sn phm ung c cha

phlorotannin v carbohydrate c hot tnh chng oxy ha.

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3.3.7 Kt qu xc nh thng s thch hp ca qu trnh ly tm. Dung dich sau khi c c cha cc mui khong v mt s tp cht khc t tan trong nc to dng cn mu trng c. V vy, ti tin hnh loi b cn bng phng php ly tm. xc nh thi gian ly tm thch hp, ti tin hnh th nghim nh sau: mi mt th nghim ly 300 ml em ly tm ti cc thi gian khc nhau l 5 pht; 10 pht; 15 pht; 20 pht v 25 pht vi tc 5000 vng/pht. Cc mu em i nh gi cm quan v kt qu thu c th hin bng 3.15. Bng 3.15 Kt qu nh gi cm quan dch chit.
Mu Thi gian (pht) Mu sc 1 5 Mu vng nu Cn cn l trong lng trong dch chit 2 10 Mu vng nu Cn cn l lng trong dch chit nhng gim ng k 3 15 Mu vng nu Dch trong khng cn cn l lng trong dch chit 4 20 Mu vng nu Dch trong, lng cn khng i 5 25 Mu vng nu Dch trong, lng cn khng i

Nhn xt: T kt qu nghin cu (bng 3.15) ta thy trong qu trnh ly tm, lng tp cht trong dch chit ph thuc vo thi gian ly tm. Thi gian cng di, lc ly tm cng c iu kin tc ng ln ton b cc phn t trong dch rong, kh nng phn tch cc tp cht cao nn lng tp cht tch ra cng nhiu, dch cng trong v sng hn. Nhng nu cng ko di thi gian ly tm, s tc ng ca lc ly tm n cc thnh phn trong dch t n trng thi n nh v lng tp cht trong dch rong lc ny khng cn nhiu nh lc u nn chng s thay i t hoc khng thay i. C th nh sau: Dch chit sau khi ly tm trong 5 pht vn cn nhiu cn l lng, lng tp cht tch ra t v mu vn km sng. Khi ly tm trong 10 pht, dch chit trong hn, lng tp cht tch ra nhiu hn nhng vn cn mt lng cn nht nh l lng trong dch chit. Khi tin hnh ly tm trong thi gian 15 pht ta thy dch chit trong, lng tp cht lng ht, mu dch sng r rt.

67

Tuy nhin khi ko di thi gian ly tm trong khong 20 25 pht quan st k ta thy dch trong v sng nhng lng tp cht tn ti trong dch vn khng thay i. Do vy, dch chit thu c trong hn v sng mu hn, ng thi trnh lng ph thi gian v nng lng ta chn thi gian thch hp nht l 15 pht lm thng s c nh cho qu trnh ly tm. 3.3.8 xut quy trnh chit phlorotannin t rong Sargassum Serratum. S quy trnh:

Rong Ra sch Ct nh Trch ly Lc

Dung mi: nc pH= 2 T l NL/DM :1/20 Nhit : 600C Thi gian :100 pht

C c (500C; 40 50 pht) Ly tm (15 pht)

B Sn xut Alginate

Cht chit Bo qun

Hnh 3.7: S quy trnh chit rt phlorotannin t rong Sargassum Serratum Khnh Ha.

68

Thuyt minh quy trnh: Nguyn liu:

La chn loi rong M t tp cht, c ngun gc v c thu mua ng cch. Rong kh cn c bo qun trong iu kin thch hp, trnh ht m, trnh tip xc trc tip vi nh sng mt tri. V khi mt s thnh phn nhy cm ca rong vi mi trng s b phn hy gy hao ht hoc lm bin tnh gi tr ca chng. Do rong c ng trong bao b nilon v di bng ti. Ra sch: Mc ch: loi b tp cht bm dnh trn rong nguyn liu. Tin hnh ra rong bng nc sch ca nh my thnh ph. Thao tc ra phi nhanh v sch, ng thi trnh lm rong b dp nt khi ra gy tn tht cht tan theo nc. Ct nh: Rong sau khi ra c mang i ro s b, sau ct nh khong 1 3 mm. y l kch thc ph hp cho qu trnh chit. C th dng ko ct theo kch thc trn. Trch ly: Rong ct nh c cho vo cc l thy tinh c np vn kn, trnh hin tng dung mi bay hi trong qu trnh ngm rong. Ta tin hnh chit trong cc iu kin: dung mi chit l nc; pH= 2; t l NL/DM l 1/20; nhit chit 600C v thi gian chit l 100 pht. Thc hin cng on chit theo phng php ngm. Khi gia nhit cho qu trnh chit, s dng t sy iu chnh c nhit , ng thi phi thng xuyn lc mu trong sut thi gian chit. Lc: Lc nhm mc ch loi b cc hp cht khng tan, b ra khi hn hp chit v thu hi dch chit. Tin hnh lc iu kin phng v s dng giy lc Whatman 01. Tin hnh c c nhit 500C trong khong thi gian 30 50 pht th t c nng cht kh hoc th tch dch chit nh mong mun. Sau khi c c hu nh dung mi nc bay hi ht. S dng thit b c quay iu chnh c nhit v p sut mt cch d dng, to mi trng chn khng v khuy o cho qu trnh c c. T hn ch c nh hng ca cc yu t mi trng, nhit cao n hm lng v hot tnh cht tan cn chit.

69

Ly tm: Mt lng tp cht cn st li sau qu trnh lc v c c s c tch ht ra khi dch chit nh vo lc ly tm ca qu trnh ly tm. V th sau khi ly tm, dch lc trong hn v sng mu hn. 3.3.9 ng dng cao Phlorotannin vo qu trnh to sn phm. a) Kho st ch tiu ha l v nht ca mt s sn phm nc ngt trn th trng. Bng 3.16: Ch tiu ha l v nht ca mt s sn phm nc ngt. Sn phm Nc yn nature Nc yn Wonderfarm Nc yn Tribeco pH 6.33 6.7 6.67 nht (Cp) 36.1 33.8 30 ng tng 10.4 8.7 9.5

Nc yn Bidrico 5.7 46 9.5 Qua kho st mt s sn phm nc ngt , ti c nhn xt nh sau: pH= 5.7 6.7, nht t 30 46 (Cp) v hm lng ng t 8.7% 10.4%. b) Kho st t l dch cao c c nc. Bng 3.17: Kt qu nh gi cm quan dch chit theo t l dch cao b sung. STT 1 2 3 4 5 6 7 8 9 Dch c : nc 1/100 1.5/100 2/100 2.5/100 3/100 3.5/100 4/100 4.5/100 5/100 Cm quan mu Vng nht Vng nht Vng hi sng Vng sng Vng sng Vng m Vng m Vng sm Vng sm Cm quan mi Thm nh Thm nh Thm nh Thm nh Thm Thm Thm Thm m Rt m

70

T l b sung dch cao phlorotannin/ nc ung cng cao th hm lng phlorotannin trong nc ung cng tng. Tuy nhin mu sc ca sn phm li sm, mi v gt v ngc li. Do , da vo kt qu bng trn ti chn t l b sung cao phlorotannin vo ung l 3/100. Chn t l ny to cho sn phm sng ng v mu sc, hp dn v mi v. c) Kho st t l ng b sung. Bng 3.18: Bng nh gi cm quan dch chit theo t l ng phi ch STT 1 2 3 4 5 6 Hm lng ng 4% 5% 6% 7% 8% 10% Cm quan v Qu nht Nht Ngt va Ngt va Ngt Rt ngt

ng cho vo sn phm nhm to v ngt cho sn phm. Hm lng ng cng cao th sn phm cng ngt. Trong bi lun vn ny, sn phm ung rong bin ng chai vi mc ch gii kht cn phi tt cho sc khe. Nu ta phi ch hm lng ng qu cao th s lm cho sn phm c v ngt gt, mt hng v c trng, v nh hng n gi thnh sn phm, n cng on thanh trng v n th hiu ca ngi tiu dng. Nhng nu qu thp th sn phm c v nht, trng thi khng tt, s khng hp dn. Do vy, t kt qu kho st cc sn phm nc ngt v t bng trn, ti chn t l ng phi ch thch hp l 8%. d) Kho st ch kh trng. Thc phm l mi trng dinh dng tt nht cho s pht trin ca vi sinh vt. Cc loi vi sinh vt pht trin mnh trong cc mi trng c acid hay pH khc nhau.

71

Nc ung rong bin ng chai cha acid, pH = 5.7 < 6 (sn phm t chua). mi trng ny vi sinh vt bn nhit, kh b tiu dit hn khi thanh trng.V vy sn phm cn thanh trng nhit t 1000C tr ln. Trong qu trnh lm th nghim vic kim tra ch thanh trng gp nhiu kh khn nn ti chn ch thanh trng 1000C. T , kho st thi gian gi nhit thanh trng sn phm ti 3 mc thi gian: 5 pht, 10 pht v 15 pht. Qua thi gian bo qun 2 tun, ti tin hnh kim tra ch tiu vi sinh, chn thi gian thanh trng ti u nht. Bng 3.19: Kt qu ch tiu vi sinh theo thi gian thanh trng. Nhit thanh trng (1000C) Ch tiu vi sinh TPC (CFU/g) 5 pht Khng pht hin 10 pht Khng pht hin 15 pht Khng pht hin

Da vo kt qu trn, ti chn ch thanh trng 1000C, gi nhit trong 5 pht. Ti ch ny, tiu dit c ht ton b vi sinh vt gy h hng sn phm, ng thi to mu sc, mi v sn phm hi ha, trng thi ng nht, c trng cho sn phm nc ung rong bin. Vy cng thc thanh trng: 10- 5- 10 1000C e) Bng m t sn phm: nc rong bin ng chai (300 ml) sau khi b sung dch cao phlorotannin. Bng 3.20: Bng m t sn phm nc rong bin ng chai c b sung cao phlorotannin. Trng thi Sn phm trong, ng nht, khng bt. Mu sc Mu vng c trng cho sn phm: tng i sng ng v hp dn Mi V

Mi thm t nhin Ngt du c trng ca sn phm rong cho sn phm, kh bin. hi ha gia v v mi, khng l khuyt tt, kh hp dn. Vi ti ny, ti chn bao b thy tinh do thy tinh tr vi tc dng ca

thc phm, c trong sut cao, hnh thc p, khch hng c th nhn thy c

72

mu sc ca sn phm, c th ti s dng c. Loi chai thy tinh c s dng c th tch l 300 (ml). f) Kt qu phn tch thnh phn ca sn phm sau khi b sung dch cao

phlorotannin. Bng 3.21: Bng phn tch cc thnh phn ca sn phm sau khi b sung cao phlorotannin. Thnh phn Phlorotannin Carbohydrat m Iod g) Kim tra ch tiu vi sinh vt trong sn phm. Bng 3.22: Kt qu kim tra ch tiu vi sinh vt trong sn phm. Tiu chun cho php 102 10 Khng c Khng c Khng c Khng c Khng c 10 Hm lng (mg/ 100ml) 27 40.5 6.3 2.4

STT

Ch tiu kim nghim TSVSVHK Coliforms E. coli S. aureus Streptococcuss faecalis P. aeruginosa Cl.perfringens TSBTNM-M

n v

Kt qu

Phng php th

1 2 3 4 5 6 7 8

Kl/ml Kl/ml Kl/ml Kl/ml Kl/ml Kl/ml Kl/ml Kl/ml

0 0 0 0 0 0 0 0

TCVN 4884:2005 TCVN 6848:2007 TCVN 6846:2007 TCVN 4830-1:2005 TCVN 6189-2:1996 3347/2001/Q-BYT TCVN 4991:2005 TCVN 5166:1990

73

Ghi ch: Kl: khun lc. BT: bo t. TSVSVHK: tng s vi sinh vt hiu kh. TSBTNM-M: tng s bo t nm men, nm mc. h) Kt qu nh gi cht lng sn phm: nc ung rong bin ng chai. PHIU KT QU NH GI CHT LNG SN PHM Php th cho im cht lng TCVN 3215 79 Sn phm: nc ung rong bin ng chai Ch tiu cht lng Trng thi Mu sc Mi V A 4 4 4 5 im cc thnh vin Tng s im B 5 5 5 4 C 4 4 4 4 D 5 5 4 4 E 5 4 5 4 F 5 4 4 5 G 4 5 4 4 32 31 30 30 Ngy th: 10-06-2012 im TB 4.57 4.43 4.29 4.29 H s quan trng 0.8 0.8 1.2 1.2 im c trng lng 3.66 3.54 5.15 5.15 17.5

Tng im

Kt qu: Sn phm xp loi: kh (theo TCVN 3215 79)

74

KT LUN V XUT KIN

1. Kt lun Qua thi gian thc hin ti, ti rt ra mt s kt lun nh sau: xy dng c quy trnh trch ly phlorotannin t rong Sargassum Phlorotannin tn ti ch yu trong t bo ca cy rong v l thnh phn c Serratum ca b bin Nha Trang- Khnh Ha. nhiu gi tr trong y hc, thc phm, m phm, Nu qu trnh thu hi, bo qun lm cy rong b dp nt s rt d gy tn tht mt lng phlorotannin nht nh. Do , ti la chn nguyn liu c ngun gc kh v c bo qun trong ti nylon v trong bng ti. Phlorotannin ha tan trong cc dung mi phn cc, s dng dung mi nc, phng php ngm chit thnh phn ny va n gin, t hiu qu cao, ng thi tit kim c ha cht s dng v thi gian thc hin qu trnh. Hm lng phlorotannin trong rong Sargassum Serratum ca b bin Nha Xc nh c cc thng s k thut cho qu trnh trch ly, l: Dung mi s dng pH T l nguyn liu/dung mi Nhit trch ly Thi gian trch ly : nc : 2.0 : 1//20 : 600C : 100 pht Trang- Khnh Ha chim khong 0.8%.

Xc nh c ch c c ph hp, c c trong iu kin chn khng Xc nh c thi gian ly tm l 15 pht. Vic ng dng cao chit phlorotannin vo trong vic sn xut nc ung

vi: nhit c 500C trong khong thi gian 30 50 pht.

rong bin ng chai tng i tt. Qua vic cm quan nh gi cht lng th sn phm t loi kh. 2. xut kin. Mc d c gng trong qu trnh lm ti, song do thi gian v iu kin nghin cu c hn nn ti vn cn nhiu hn ch v thiu st. Do vy, ti xin xut mt s kin sau:

75

Cn nghin cu thm v kh nng khng oxy ha v nhng tc dng

khc ca cc cht c trong rong. Cn nghin cu thm cc phng php thu hi, bo qun rong ti

nguyn liu hn ch tn tht cc thnh phn qu trong rong , ng thi m bo nguyn liu chit ngay c trong thi gian tri v. Tip tc nghin cu quy trnh, c bit l trong cng on chit (nh

nghin cu chit phlorotannin vng bin c iu kin sng ca rong khc nhau, thc hin chit trong cc mi trng pH khc nhau, dung mi khc nhau,). V vn ny t c nghin cu trong nc. Cn nghin cu thm qu trnh x l v tinh ch dch trch ly, qu

trnh lnh ng v ng kh dch c nhm ko di thi gian bo qun v thu c sn phm c cht lng tt hn, Nghin cu quy trnh chit phlorotannin t i tng rong khc. Cn nghin cu thm qu trnh thu nhn v x l cao chit thnh ch

phm dng bt. T thun li cho vic thng mi ha v s dng b sung vo cc loi sn phm khc. Cn nghin cu kh nng ng dng rng ri ca ch phm vo trong

cc lnh vc nh: thc phm, dc phm, y hc, m phm,

76

MT S HNH NH MINH HA

Hnh 1: Cc l mu rong v b rong sau khi lc.

Hnh 2: Mu dch chit sau khi b sung cao phlorotannin.

77

TI LIU THAM KHO

1. Trn Th Luyn, Minh Phng, Nguyn Anh Tun, Ng ng Ngha (2004), Ch bin rong bin, NXB Nng Nghip TP H Ch Minh. 2. Nguyn Hu i (1997), Rong M Vit Nam ngun li v s dng, NXB

Nng Nghip TP H Ch Minh. 3. Minh Phng, ng Vn Hp (1997), Phn tch kim nghim sn phm

thy sn, Trng i Hc Thy Sn Nha Trang. 4. Khoa ha thc phm v cng ngh sinh hc, Cc qu trnh cng ngh c bn

trong sn xut thc phm, Trng i hc Bch Khoa H Ni. 5. Nguyn Hi H (2004), Nghin cu trch ly polyphenol t tr Camellia

sinensis (L), Lun vn Thc s, i hc Bch Khoa TP H Ch Minh. 6. ng Xun Cng (2009), Nghin cu thu nhn dch chit c hot tnh

khng khun t rong Nu Dictyota Dichotoma Vit Nam, Lun vn Thc s, i hc Nha Trang. 7. 8. Mt s n tt nghip ca anh ch kha trc. Riitta Koivikko (2008), Brown algal phlorotannins improving and applying

chemical methods, Annales Universitatis Turkuensis. 9. Rangan M. A. and Glombitza K. W. 1986, Phlorotannin, Brown algal

polyphenols, Prog. Phycol. Res, 4: 1290 241. 10. Yong Li, Zhong- Ji Qian, Bomi Ryu, Sang- Hoon Lee, Moon- Moo Kim, Se-

Kwon Kim, Chemical components and its antioxidant properties in vitro: Anedible marine brown algal, Ecklonia cava, Bioorg. Med. Chem. (2009),

doi:10.1016/j.bmc.2009.01.031. 11. Mayalen Zubia, Daniel Robledo, Yolanda Freile- Pelegrin, Antioxidant

activities in tropical marine macrroalgae from the Yucatan Peninsula, Mexico, J Appl PHYCOL (2007) 19:449-458. 12. Koki Nagayama, Yoshitoshi Iwamura, Toshiyuki Shibata, Izumi Hirayamal

and Takashi Nakamura, Bactericidal activity of phlorotannins from the brown algal, Ecklonia kurome, Journal of Antimicrobial Chemotherapy (2002) 50, 889- 893. [Cng b nm 2002]

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13.

TL: Sang- Hoon Lee, Li- Yong, Fatih Karadeniz, Moon- Moo Kim, Se-

Kwon Kim, Alpha- glucosidase and alpha- amylase inhibitory activities of phlorotannin derivatives from Ecklonia cava, Journal of Biotechnology 136S (2008) S577- S588. 14. Moon- Moo Kim, Quang Van Ta, Eresha Mendis, Niranjan Rajapakse, Won-

Kyo Jung, Hee- Guk Byun, You- Jin Jeon, Se- Kwon Kim, Phlorotannins in Ecklonia cava extract inhibit matrix metalloproteinase activity, Life Sciences 79 (2006) 1436- 1443. 15. Murat Artan, Yong Li, Fatih Karadeniz, Sang- Hoo Lee, Moon- Moo Kim,

Se- Kwon Kim, Anti- HIV- 1 activity of phloroglucinol derivative, 6,6- bieckol, from Echlonia cava, Bioorganic & Medicinal Chemistry 16 (2008) 7921- 7926. 16. Quang- To Le, Yong Li, Zhong- Ji Qian, Moon- Moo Kim, Se-Kwon Kim,

Inhibitory effects of polyphenols isolated from marine alga Ecklonia cava on hictamine release, Process Biochemistry 44 (2009) 168- 176. 17. Nelson T.E., Lewis B.A Separation and characterization of the soluble and

insoluble components of insoluble laminaran // Carbohydr. Res. 1974.- Vol.33.P. 63- 74. 18. Maria E.R. Duarte, a Marc A. Cardoso, a Miguel D. Noseda, a, l Alberto

S.Cerezo (2001), Structural studies on fucoidans from the brown seaweed Sargassum Stenophyllum, Carbohydrate Research 333, pp 281- 293. 19. Google.com.vn

PH LC
Ph lc 1: Xc nh hm lng m bng phng php sy [3]. a) Nguyn l:

Dng nhit cao lm bay hi ht nc trong mu th, sau da vo hiu s khi lng ca mu th trc v sau khi sy kh, t tnh c hm lng nc trong thc phm (%). b) c) Dng c v ha cht: T sy iu chnh c nhit . Cn phn tch, chnh xc 10-4 g. Cc sy bng thy tinh. a thy tinh. Bnh ht m. Tin hnh:

Sy cc n khi lng khng i: Cc c ra sch, p kh, sy nhit 100 105 0C trong khong 1 gi, sau ly ra lm ngui trong bnh ht m ri mang i cn v sy tip nhit trn, lm ngui trong bnh ht m v cn n khi no gia hai ln lin tip sai khc khi lng khng qu 5.10-4 g l c (sy n khi lng khng i). Cn chnh xc mt lng rong ct nh (khong 10 g) vo cc sy kh n khi lng khng i. Dng a thy tinh nh ti mu ri dn u mu trn y cc. Chuyn cc vo t sy, sy 60 800C trong 2 gi. Sau nng nhit ln 100 1050C, sy lin tc trong 3 gi. Ch , trong qu trnh sy c sau mt gi o mu mt ln. Ly mu ra ngui trong bnh ht m, sau mang cn trn cn phn tch ri sy tip nhit 100 1050C n khi lng khng i nh trn. d) Tnh kt qu:

m ca rong c tnh theo cng thc:

X G 1 G 2 * 100 G 1 G (%)

Trong : X : m (hm lng nc) ca rong (%). G1: Khi lng cc sy v mu th trc khi sy (g). G2: Khi lng cc sy v mu th sau sy (g). G : Khi lng cc sy (g). Ph lc 2: Xc nh hm lng protein theo phng php Kjeldadl. T hm lng nito nh lng c theo phng php Kjeldadl, nhn vi h s 6.25 s tnh c hm lng protein trong mu rong. Phng php Kjeldadl [3]: a) Nguyn l:

xc nh m tng qut trong thc phm ngi ta dng H2SO4 m c v cht xc tc c bit l hn hp CuSO4 v K2SO4 theo t l 1/10 v c ha; mc ch chuyn ton b nito trong thc phm v dng mui (NH4)2SO4. Sau dng NaOH c y NH3 ra v dng hi nc li cun NH3 ra khi thit b chng ct vo cc hng cha H2SO4 1.1N d, cui cng dng NaOH 0.1N chun li H2SO4 d. b) Cch tin hnh: Ly chnh xc 2 g mu rong ct nh cho vo bnh Kjeldadl. Thm 4 g hn

hp cht xc tc CuSO4 v K2SO4 v 20 ml H2SO4 m c nghing trn bp in un t t. Trong qu trnh v c ha mu sc ca mu chuyn t mu nu en sang mu vng ri xanh trong hoc khng mu l c. Ch : Trong qu trnh v c, nu mu cha t n mu xanh trong m dung

dch b cn th ly bnh ra ngui v thm vo 5ml dung dch H2SO4 m c ri tip tc v c. Khi v c ha mu, trnh hin tng mu si qu mnh b bn ra

ngoi gy sai s, phi v c trit hon ton. Ra thit b chng ct: Tin hnh sc ra thit b chng ct v kim

tra cc khp ni phi m bo sch v kn.

Chun b cc hng: Ly cc thy tinh 250 ml sch cho vo 25 ml dung dch

H2SO4 0.1 N v vi git metyl 0.1%. t cc hng ngp di u ng sinh hn ca thit b chng ct. Chng ct v chun : cho mu v c ha vo bnh chng ct ca thit

b, trng bnh Kjeldadl vi ln bng nc ct ly ht mu th v c ha, dch trng cng chuyn c vo bnh chng ct. Thm vi git phenolphthalein 1% v cho t t dung dch NaOH 40% cho ti khi c mu (hoc tm ), trng phu bng nc ct, y kn cc khp ni v tin hnh chng ct li cun NH3 bng hi nc cho n khi ht NH3 trong bnh chng ct (th bng giy o pH). nh lng trc tip NH3 bay ra bng H2SO4 0.1N d trong cc hng, sau dng NaOH 0.1N chun li lng H2SO4 0.1N d trong cc hng n khi dung dch chuyn t mu sang mu vng nht l c. c) Tnh kt qu:

Hm lng m ton phn tnh theo cng thc sau:

Ntp 0 . 0014 * ( A B ) * 100 P (%)

Trong : A: s ml H2SO4 0.1N dng trong cc hng B: s ml NaOH dng trong chun . P: khi lng mu th (mu rn) (g). 0.0014: s gam nito tng ng vi 1 ml H2SO4 0.1N. Hm lng protein trong rong c tnh theo cng thc sau: Xp = Ntp* 6.25 (%) Ph lc 3: Xc nh hm lng lipid tng bng phng php Bligh v Dyer hn hp chloroform/methanol a. Nguyn l:

Gii phng lipid t cc hp cht vi protein v glucid nh thy phn bng chloroform (CHCl3) mi trng c cn (CH3OH). b. Dng c ha cht: S dng dng c v ha cht thng thng ca phng th nghim.

c.

Tin hnh:

Cn 50- 100 gam mu rong hay b rong c nghin nh trong my xay, ri em ngm chit trong CHCl3- CH3OH t l 1: 2 (100 ml CHCl3: 200 ml CH3OH) v siu m trong 6 gi (chit 2 ln). B sung 100 ml CHCl3, sau thm nc vo lc u. Phn lp ly phn dung dch pha di , ra li bng nc 2 ln, sau em lm khan bng Na2SO4. Dch chit thu c em c ct loi dung mi trn my quay ct chn khng 400C v p sut 25 tor. Hm lng du bo thu c sau khi cn trn cn phn tch Sortorius analytic (10-4) v c tnh ton theo phn trm khi lng so vi mu ban u. d. Tnh kt qu:

Hm lng lipid (%) theo cng thc sau:

A P * 100 G (%)

Trong : A: hm lng lipid trong mu th (%) P: khi lng lipid em cn (g) G: khi lng mu th (g). Ph lc 4: Xc nh hm lng Iod theo phng php chun [3]. a) Nguyn l: Iodua (I-) c oxy ha bng nc Brom bo ha thnh Iodat (IO3-), Iodat kt hp (I-) trong mi trng acid gii phng I2 t do. Chun bng Na2S2O3 vi ch th h tinh bt. T tnh ra lng Iod trong thc phm. Cc phn ng chnh: KI + 3 Br2 + 3 H2O = KIO3 + 6 HBr KIO3 + 5 KI + 2 H3PO4 = 3 I2 + 2K3PO4 + 3 H2O I2 + Na2S2O3 = 2 NaI + Na2S4O6 b) Tin hnh Cn 20 gam mu cho vo cc nung. Trn u vi 10g bt MgO khng ch nhit trnh Iod thng hoa. Nung trong 2h nhit ti a l 3500C. Ha tan tro bng nc ct (10- 15ml) ri lc trn giy lc khng tn, ly dung dch. Phn tro

cn li v giy lc nung li nhit 450- 5000C. Sau ha tan bng nc ct, lc trn giy lc khng tn. Nu cn cn en th tip tc nung li ln 3 thnh tro trng (phi nung nhiu ln trnh Iod thng hoa). Dch lc dn chung vo bnh nh mc 50 ml, thm nc cho mc c dung dch th. Cho vo bnh tam gic c nt mi kn: Dung dch th Nc Brom bo ha lm lnh n 100C. Cho thm vo bnh tam gic: H3PO4 (d = 1.3): 1ml KI tinh th: 1g y np, lc u, yn trong bng ti 15 pht ( phn ng hon ton). Sau m np thm 1 ml h tinh bt 1% dung dch c mu xanh tm. Chun bng Na2S2O3 0.01N ti khi mt mu xanh tm. c th tch Na2S2O3 0.01N dng ht. T tnh ra hm lng Iod theo cng thc:
X 1 N * 0 . 21 * F * 100 P (%)

: 10ml : 3 ml

Dung dch H2C2O4 10% (acid oxalic) : 6 git. un cch thy bnh tam gic khong 10 pht ui Brom d. ngui ri

Trong : X1 : hm lng Iod trong mu phn tch (%) N F P : s ml Na2S2O3 0.01N dng chun (ml) : h s pha long : khi lng mu phn tch (g) 0.21 : s mg I2 tng ng 1ml Na2S2O3 0.01N.

xc nh hm lng Iod trong dch chit cng tin hnh nh trn t khu cho vo bnh tam gic. Ph lc 5: Xc nh tro ton phn [3]. a) Nguyn l Dng sc nng (550- 6000C) nung chy hon ton cc cht hu c. Phn cn li em cn v tnh ra hm lng tro ton phn trong mu phn tch. b) Dng c, vt liu v thuc th

 c)

Chn nung bng s. n cn hay bp in. L nung iu chnh c nhit (n 550- 6000C) Cn phn tch. Bnh ht m, pha di cht ht m. H2O2 10 th tch hoc HNO3 m c. Tin hnh

Nung chn s ra sch l nung ti n 550- 6000C n trng lng khng i. ngui bnh ht m v cn phn tch chnh xc n 10-4 g. Cho vo chn khong 5 gam cht th. Cn tt c cn phn tch vi chnh xc nh trn. Cho tt c vo l nung v tng nhit t t cho n 550- 6000C. Nung cho n cho trng , ngha l loi ht cc cht hu c thng thng khong 6-7 gi. Trng hp cn tro en, ly ra ngui, cho thm vi git H2O2 10 th tch hoc HNO3 m c v nung li cho n tro trng. ngui trong bnh ht m v cn cn c chnh xc nh trn. Tip tc nung thm nhit trn trong 30 pht ri ngui trong bnh ht m v cn, lp i lp li thao tc ny cho ti trng lng khng i. Kt qu gia 2 ln nung v cn lin tip khng c cch nhau qu 0.0005 gam. d) Tnh kt qu
G2G * 100 (%) G1 G

Hm lng tro theo phn trm (X1) tnh bng cng thc:
X1

Trong : X1: Hm lng tro trong mu phn tch (%) G1: khi lng chn nung v mu (g) G : khi lng chn nung (g) G2 : khi lng chn nung v tro trng (g) Ch : Khi chn nung cn nng vo bnh ht m, nh np h m lc u hoc m vi khng kh trn np bnh ht m trnh khng kh nng n ra y lm bt v np bnh.

Ph lc 6: Xc nh hm lng x th theo Crudefibre v nh lng Carbohydrat. xc nh x th, vic nh gi c x l da trn rong nhin liu loi m v cht bo vi acid long 1.25 %, sau vi kim 1.25%. Nh vy, 2 g rong nhin liu loi m v cht bo c x l vi 200 ml dung dch H2SO4 1.25%. Sauk hi lc v ra, b c x l vi dung dch NaOH 1.25%. B c lc, ra vi nc nng v sau vi HNO3 1% v ra li vi nc nng. B c sy kh 2 gi 1300C 20C, ngui trong bnh ht m v cn (W2). Nung 30 pht 6000C 150C, ngui trong bnh ht m v cn (W3).
CrudeFiger W 2 W 3 * 100 (%) W1

Trong : W1: lng mu th Xc nh hm lng carbohydrate: % Carbohydrat = 100 (m + protein + cht bo + tro + x) Ph lc 7: Xc nh hm lng Fucoidan Lng sinh khi rong nghin mn (100g) c x l nhit phng vi hn hp MeOH- CHCl3- H2O = 4:2:1 loi b cht mu, lc sy kh chn khng cho ra mt lng sinh khi rong kh cht bo. Nguyn liu ny v dung dch CaCl2 2% (5250ml) c khuy trn c hc ti 850C trong 5 gi. a 80 ml dung dch hexadecyl trimethylammonium bromide nng 10% trong nc vo trong cc dung dch chit c gom chung li. Ta to thnh c ly tm, ra vi nc khuy vi dung dch NaI 20% trong cn (5150ml) trong vng 2- 3 ngy nhit phng, ra vi cn v ha tab trong nc. Dung dch c thm tch v ng kh nhanh thu c fucoidan th (F) di dng mui Na+. Cn trn cn phn tch Sortorius analytic (10-4) v c tnh ton theo phn trm khi lng so vi mu ban u. Ph lc 8: Xc nh hm lng Alginate. Cn khong 10 gam rong kh, ngm rong trong 500 ml formaldehyt 1%, khuy nh trong 12 gi. Sau ra sch rong bng nc ct. Tip tc ngm rong trong H2SO4 0.2N trong 4 gi ri li ra sch bng nc ct. Chit alginate bng Na2CO3 1% trong 12 gi, c khuy nh. Lc thu dch lc. Ta bng cn 950 theo t

l dch chit: cn l 1: 2, lc thu ta, ra ta bng acetone, sy kh 400C. Cn sn phm v tnh thnh phn phn trm. Hm lng alginate (%) tnh theo cng thc:
A P * 100 G (%)

Trong : A: hm lng alginate (%) P: khi lng alginate em cn (g) G: khi lng mu th (g) Ph lc 9: Xc nh hm lng phlorotannin trong dch chit. Hm lng phlorotannin trong dch chit c xc nh bng phng php so mu, dng thuc th Folin- Ciocalteu bc sng 750 nm. a) Nguyn l: Oxi ha ton b lng phlorotannin trong dch chit bng dung dch FolinCiocalteu (hn hp acid phosphotungstic v acid phosphomolyblic). Cc acid ny s b kh thnh Vonfram (W8O23) v oxyt molipden (Mo8O23) c mu xanh. Mu xanh c hp th nhiu nht bc sng 750 nm. b) Dng c: c) 10%. Sau t cc ng nghim trong bng ti nhit phng khong 1 gi. Sau khong thi gian trn quan st thy mu xanh en xut hin v tin hnh so mu bng my quang ph UV- Vis bc sng 750 nm, ghi hp th v tnh ton xc nh c hm lng phlorotannin c trong mu th. My quang ph UV-Vis. Pipet Bnh nh mc. ng ong. Cc ng nghim. Tin hnh: Dng c, ha cht:

Ly 1ml dch chit dung mi + 1 ml Folin- Ciocalteu 10% + 2 ml Na2CO3

Ch : Trc khi tin hnh th nghim ta phi pha long mu nu nh cn thit. Ph lc 10: Xc nh hm lng Carbohydrat trong dch chit. Ly 0.5 ml dung dch cn xc nh c nng trong khong 10- 100 g/l. Thm 1 ml dung dch cn: phenol (1:8) lc u, thm 5 ml H2SO4 m c ri em un cch thy ti nhit 600C trong 10 pht, ngui. em dung dch trn o mt quang 484 nm, cht chun dng galacto. Ch : Trc khi tin hnh th nghim ta phi pha long mu nu nh cn thit. Ph lc 11 Bng 3.3: Kt qu xc nh nh hng ca pH n kh nng trch ly phlorotannin. P(mg/Vdch trch ly t 10 (g) nguyn liu) 26.26 21.64 22.03 19.18 18.44 18.76 18.92 19.39 17.43 17.29 15.53

pH 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 7.0 8.0

OD 0.254 0.201 0.221 0.179 0.184 0.188 0.190 0.196 0.194 0.192 0.147

pha long 10 10 10 10 10 10 10 10 10 10 10

V dch trch ly (ml) 178 175 165 170 160 160 160 160 145 145 160

Ph lc 12 Bng 3.4: Kt qu xc nh nh hng ca pH n kh nng trch ly carbohydrate. pH 1.0 1.5 2.0 2.5 3.0 3.5 4.0 5.0 6.0 7.0 8.0 OD 0.339 0.337 0.330 0.312 0.310 0.301 0.304 0.305 0.316 0.313 0.302 pha long 5 5 5 5 5 5 5 5 5 5 5 V dch trch ly (ml) 178 175 165 170 160 160 160 160 145 145 160 carbohydrate (g/ Vdch trch ly t 10 (g) nguy n liu) 70975.5 69343.75 63937.5 62050 58000 56200 56800 57000 53650 53106.25 56400

Ph lc 13 Bng 3.5: Kt qu xc nh nh hng ca t l nguyn liu: dung mi n kh nng trch ly phlorotannin. T l 1: 5 1: 10 1: 15 1: 20 1: 25 1: 30 1:40 1:50 OD 0.556 0.376 0.242 0.218 0.170 0.153 0.119 0.072 pha long 10 10 10 10 10 10 10 10 V dch trch ly (ml) 18 67 105 165 210 250 310 410 P(mg/Vdch trch ly t 10 (g) nguyn liu) 5.37 14.06 15.10 21.78 22.76 25.00 25.81 24.64

Ph lc 14 Bng 3.6: Kt qu xc nh nh hng ca t l nguyn liu: dung mi n kh nng trch ly carbohydrate. T l 1: 5 1: 10 1: 15 1: 20 1: 25 1: 30 1: 40 1: 50 OD 1.103 0.541 0.518 0.310 0.266 0.283 0.264 0.167 pha long 5 5 5 5 5 5 5 5 V dch trch ly carbohydrat(g/ Vdch trch ly (ml) t 10 (g) nguyn liu) 18 67 105 165 210 250 310 410 24367.5 43633.75 65362.5 59812.5 64575 82187.5 94550 75337.5

Ph lc 15 Bng 3.7: Kt qu xc nh nh hng ca nhit n kh nng trch ly phlorotannin. Nhit (0C) 40 50 60 70 80 90 100 OD 0.174 0.184 0.243 0.250 0.233 0.224 0.205 pha long 10 10 10 10 10 10 10 V dch trch ly (ml) 165 165 165 165 165 165 165 P(mg/Vdch trch ly t 10 (g) nguyn liu) 18.21 19.02 23.82 24.38 23.00 22.27 20.73

Ph lc 16 Bng 3.8: Kt qu xc nh nh hng ca nhit n kh nng trch ly carbohydrate. Nhit (0C) 40 50 60 70 80 90 100 OD 0.291 0.295 0.269 0.301 0.355 0.370 0.319 pha long 5 5 5 5 5 5 5 V dch trch ly (ml) 165 165 165 165 165 165 165 carbohydrat(g/Vdch trch ly t 10 (g) nguyn liu) 55893.75 56718.75 51356.25 57956.25 69093.75 72187.5 61668.75

Ph lc 17 Bng 3.9: Kt qu xc nh nh hng ca thi gian n kh nng trch ly phlorotannin. Thi gian (pht) 20 40 60 80 100 120 140 160 180 200 220 240 OD 0.209 0.215 0.223 0.234 0.262 0.268 0.269 0.282 0.285 0.288 0.271 0.278 pha long 10 10 10 10 10 10 10 10 10 10 10 10 V dch trch ly (ml) 165 165 165 165 165 165 165 165 165 165 165 165 P(mg/Vdch trch ly t 10 (g) nguyn liu) 21.05 21.54 22.19 23.08 25.36 25.85 25.93 26.99 27.23 27.47 26.09 26.66

Ph lc 18 Bng 3.10: Kt qu xc nh nh hng ca thi gian n kh nng trch ly carbohydrate. Thi gian (pht) 20 40 60 80 100 120 140 160 180 200 220 240 OD 0.198 0.270 0.280 0.311 0.327 0.333 0.379 0.429 0.435 0.420 0.399 0.297 pha long 5 5 5 5 5 5 5 5 5 5 5 5 V dch trch ly (ml) 165 165 165 165 165 165 165 165 165 165 165 165 carbohydrat(g/Vdch trch ly t 10 (g) nguyn liu) 36712.5 51562.5 53625 60018.75 63318.75 64556.25 74043.75 84356.25 85593.75 82500 78168.75 57131.25

Ph lc 19 Bng 3.11: Kt qu xc nh hiu sut trch ly phlorotannin S ln Ln 1 Ln 2 Ln 3 Ln 4 OD 0.262 0.467 0.251 0.148 pha long 10 V dch trch ly (ml) 165 165 165 165 P (mg/ Vdch trch ly t 10 (g) nguyn liu) 25.36 4.20 2.45 1.61

Ph lc 20 Bng 3.12: Kt qu xc nh hiu sut trch ly carbohydrate. S ln Ln 1 Ln 2 Ln 3 Ln 4 OD 0.324 0.506 0.332 0.171 pha long 5 V dch trch ly (ml) 165 165 165 165 carbohydrat(g/Vdch trch ly t 10 (g) nguyn liu) 62700 20047.5 12870 6228.75