TAP CHl KHOA HOC A C A N G NGHf CAC T R U O N C DAI HQC K V T H U ^ T « S 6 19 - 2012
KHAO SAT KHA NANG HAP PHV vA Bi^N D 6 I TINH C H A T CCA P-CHITIN (PHAN Li^P Ttr MAI MV*C) TRONG MAI TRlTdNG KI^.M
STUDY ON THP. ABSORPTION BEHAVIORS A N D PROPERTILiS CHANGES OF p-CHlTIN IN A L K A L I ENVIRONMENT
yirtmg Ngifc Chinh', Hu^nh Thi Thiiy Trang Thanh', Trhn Thuang Qu^ng^, Tiing Thanh Danh'
' Trudng Dgi hgc Bdch Khoa-DHQG HCM ' Trudng Dgi hgc Bdch khoa Hd Ndi
Nh$n ngiy 08 thing 02 nim 2012, chip nh^n ding ngiy 6 thing 6 nim 2012
T6IVI TAT
Khd ndng hdp phg vd bidn ddi tinh chdt cua f3-chitln dugc phdn Idp tir mai mi^ dng Phan Thidt Vi^l Nam tmng mdi Irudng kidm dd dugv khdo sdt & cdc didu kidn khdc nhau nhw. ndng <Sd dung dich kidm, t^ Id. nhidt dd. vd th&i gian. Sg' bidn ddi tinh chdt cua chitin dugc xdc dinh thdng qua phd nhidu xg XRD. phd hdng ngogi FTIR vd kinh hidn vi didn td' qudt SEM, quang phd tu ngogi khd kidn UV-VIS. Kdt qud nghidn cOv dd cho thdy rdng khd ndng hdp phg dung dich cda chitin tdng khi nhidt dd gidm ho$c ndng dd kidm tdng Cdu tnic tinh thd cua {3-chitin bi phd hOy tgo thdnh chitin gel vd hdp phg dung dich nhidu nhdt & nhidt dd -4(fC v&i NaOH 4M. tyl$ 1.75(g/ml) trong 4 gi& phdn Cmg.
ABSTRACT
Absorption behaviors of B-ehitin in alkali soiutkjn under different treatment conditkins of concentration, soaking time and temperature, ratio of p-chitin to alkali si^utkjn were analyzed. The solid state structural changes of the resulting chitin were inverstigated by X-ray diffraction, FT-IR spectroscopy, SEM and UV-VIS spectrosrxipy. The results showed that the absorbency increased as temperature decreased or alkali concentration increased Maximum absorbency and establish of 0- ch'itin were achieved at ~ 4(fC with a NaOH 4M solution and the ratk) 1:75 (g^l) after soaking 4 hours.
I.GldlTHI$U
Chitin la mgt thinh phin quan trgng ciia Idp vd Cling bao bgc ben ngoii cic loai sinh v$t sdng dudi nudc vi tren c ^ . mgi polysaccharide mach thing cd nhifiu trong ty nhifin vi dugc xem nhu dan xuai cua cellulose, trong dd nhdm (-0H) d nguyen tu C(2) dugc thay the bing nhdm acetyl amino. Chmn dugc su dyng nhieu trong thyc pham, my pham. dugc pham do ed kha ning tuong hgp sinh hgc, it gay dj Lme.
khdng dgc h^i. Nghien ciru sy bien ddi tinh eluit cua chitin cd the nang cao hifiu qua su dyng chiing trong tucmg lai.
Chitin li poly (p-(l-4)-2-acetaniido-2- dcoxy-D-glucosamine) [Ij. Chitin phan l^p tir mai myc dng Phan Thiit Vi?i Nam la P-ehilin.
Dang [i-chitin [2. 3] ed cac mic xich ghep vdi nhau theo kieu song song, lifin kfit hydro ngi va ngoai trong cau Iriic p-chilin yfiu nfin nd de bi inh hudng boi cac dung mdi va cho phan irng hda hgc dfi dang hem.
p-Chiiin cd khi ning hip phy dung djch do trong mdi trudng kiem vdi cac diiu ki^n phan ung thich hgp xiy ra sy phi huy ciu triic tinh thfi vi) lien ket hydro ngi vi ngo?! trong m^ng chitin. Sy hap phy niy bao gdm hip phy v9t ly vi hda hgc nhung v i ^ phan bifit co chi hap phy eua chitin thi rat khd khin vi cho dfin nay ihl chua cd lai li^u nao cdng bo rd ring.
Khi ning hip phy dung djch anh hudng rat Idn den sy bifin ddi cau tnic chitin cung nhu lam ting tinh truong nd va dg truyen suit cua chitin.
Bai bio na> t^p trung vao vific phan tich cac yeu td inh hudng tdi khi ning hap phy dung djch ciia chitin trong mdi trudng kiem.
.\cin \ct sy bifin ddi cau true tir dang p-chitin tinh [he sang dang chitin gel.
2. P H I <)N(; PHAP 2.1. N<iu>cn li^u
p-Chilin d^ng bgt khd (VNC-2010. Bich Khoa) dugc phan lap tir mai myc dng Phan
TAP CHl KHOA HQC & C6NG NCH$ CAC TRII'ONG D^l HQC K? THUAT 4 S6 89 - 2012 Thifit Vi?t Nam, d$ im 14.4%, him lugng tro
0.001%, trgng lugng phan tu 5.1x10* (Dalton) va DA 85%. NaOH, methanol, acetone, di khdi.
2.2. Phmmg phip phfin tfeh / Phuang phdp do dg hdp phg
Cin O.lg p-chitin cho vio binh phin img, sau dd them dung djch NaOH. Thyc hi^n cic diiu ki§n thich hgp vi khuiy m^nh di khao sit sy bifin ddi tinh chit cua chitin. Kfit thiic qui trinh, dem sin phim ly tam di lo^i bd kiim du vdi tic dg ly tim li 5000(vdng/phut) trong 20phul, rua sin phim bing nude eat (pH = 7) vi tinh dg hip phy dung djeh ciia chitin theo cdngthijrc(l):
AV(g/mgchiiin) = '"'^^^~"'°^^Km (1) Vdi AV(g/100g chitin) la dg hip phy dung djch eua chitin linh trfin lOOg chitin khd, mo(g) khdi lugng P-chitin ban diu, mi(g) khoi lugng sin pham sau tie kich.
2. Phuang phdp kinh hien vi dien lu quel SEM Cac miu dugc say khd va sau dd duge phii plalin (20nm) trfin bfi mat mau trong chan khdng vi bfi mat ciia mau dugc quan sal bang kinh hiin vi difin tu quel LEO-440 su dyng mgt dienip 12 kV.
3. Phuang phdp phd hdng ngogi FTIR Phd hdng ngoai FTIR ciia mau dugc do trfin may TENSOR37, d^ng vifin nen KBr.
DA(%): miirc dg acetyl dugc xie djnh thdng qua phi hdng ngoai theo phuang phap[i,4,5], linh theo cdng thuc(2):
DAi%) = [[A^f^g/A^^^)/].33][]00-baseline] (2) Trong dd A,ess la kha nang hap thu eiia nhdm amide bae 1 va A3431 kha ning hap thu eiia nhdm hydroxy! iheo quang phd hdng ngo^i.
1.33 la he sd theo phuang phap [4], [100- baseline] la he si hieu chinh lir phd dd.
4. Phuang phdp nhieu xg XRD
Phi nhifiu xa tia X dugc do tren may D8ADVANCE-BRUKER Germany vdi gde quet 29 = 3 - 50°, sir dung tia birc xa CuKa tai 50kV. Thdng sd (leu) dugc linh tir ket qua gian
do vi kich thudc trung binh ciia tinh thi (D,p(iioi) xic djnh theo phucmg phip giin di nghifin ciru [6]. Cudng dO t?i cic dinh peak m^t ludi (IIIO, dng vdi gde qufit 2B = 20° nhiiu x?
m^nh nhit, !,„, vdi gde qufit 26 = 160 (peak vd djnh hinh nhiiu x?)) di dugc sir dyng di tinh toin IcR Iheo cdng Ihirc (3). Thdng s6 (Dip [no]) dugc tinh toin theo cdng thde (4) [7],
/..
Aio<100
" ^ " " i ' ^ ^ c o s e
(3)
(4)
Trong dd K li hing si tir 0.8-1.1, Xcu - 1.540596 (A") li dg dii sdng hire x? eua dong; Po (rad) li dO bin rgng ciia peak tinh thi, 0 (rad) li nCta gde Bragg ciia peak tinh thi [5].
5. Phuang phdp do dg truyen sudt Dg truyin suit T% (do dye) cua chitin gel dugc do tren may quang phd tir ngoai - kha kifin hieu Helios Epsilon dvdi budc sdng A. = 400.
6. Phucmg phdp do trgng lu'gng phdn tu Trgng lugng phan tu (Mw) dugc tinh theo cdng thire Mark-Houwink, vdi cic hang sd a va K d 30''C li 0.95 va 7.6x10"Ml/g ciia chitin trong N,N-dimethylacetamide/lithium chloride [8]. [tl] do nhdt theo cdng thirc (5):
[q]=KM:' (5)
3. KET QUA VA THAO LUAN
3.1. Khi nang hap phy dung dich va dg truyen sunt ciia p-chitin
Nhiet dg va ndng do kifim la mgt yfiu td anh hudng rit Idn den kha nang hip phu dung djch va do truyin suit ciia p-chitin thi hien (hinh 1). Khi ndng dg kifim ling cao thi cac lien kit hydro ndi va ngoai trong cau triic chitin de dang bi eii diit. cac lien kit nay sg kit hgp vdi cac ion OH" cd trong dung djch kiem hinh lhanh phan til nudc. cac phan tu nay se khuech tan dinh chit trong mang chitin. Nhu vay chinh cac ciu ndi hydro bi dul cang nhifiu cang lam tang kha nang hip phy dung dich ciia chilin. Nhung khi ndng dp kiem tang qua cao Iren 5M va nhiel dp cao Iren eO^C cd kha nang dang P-chitin
TAP CHl KHOA HQC & C6NG NGH$ CAC TRlfftNG DAI HQC K 9 THUAT • S 6 89 - 2012
chuyin thinh a-chitin, lifin kit hydro n$i vi ngo^i trong a-chitin bin hon n<n khd ddt ra vi m$t phin chitin bj deacetyl thinh chltosan do dd khi ning hip phy dung djch vi dd truyin suit cCia chitin li khdng ding ki.
Khi h? nhiet dd lit 30"C xuing -I0''C»
trong mdi trudng kiim khi ning hip phy dung djch vi d^ truyin suit cCia chitin bit diu tfing d$t biin, cd hi^n tirgng phi ciu triic llnh Ihi ehitin vi chuyin thinh d^ng chitin get. Nhift dd ty 1$ nghjch vdi khi ning hip phy dung djch cua chitin, nhift d$ cing giim Iht khi nfing hip phy dung djch ciia chitin cing ting vi vdi nhifl
dd tic kich 40°C trong NaOH 5M thi khi nfing hip phy dung djch c6a chitin li cao nhit li 97Sg trtn tOOg chitin khd. V9y nhift d$ thich hgp dk tic kich p-chitin trong mfli trudng kiim li -40°C.
Ngoii y ^ ti nhift dfl. ning dd thi ty If chitin/dung djch kiim cQng inh hudng dfing ki din khi n&ng hip phy dung djch cda diitin. Khi tfing ty If chitin/dung djch kiim \tn thi chitin truong nhiiu hem trong dung djch. Diiu niy thi hifn rd khi sir dyng NaOH 4M (hinh 2). Theo kit qui khio sit ty If l:75(g/mt) li diiu kifn thich hgp di tic kich p-chitin.
Hinh 1. Dd thi dnh hudng ddng thai nhi4l dd vd ndng dg kiem tdc kich den khd ndng hdp phg dung dich vd dg truyen sudl eua P-chitin
_ 1500.00
§ 1000.00
Hmh 2. Dd thi dnh hudng ddng thai ly i(' vd ndng Hinh 3 Dd thi dnh hudng thdi gian tdc kich den dg kiem tdc kich den khd ndng hdp phg dung dich khd ndng hdp phg dung dich cua ^-chilin a -4(fC
vdi NaOH 4M
^ 6 0
NiUHiM) NDng(loNaOH(M) j
Hinh 4 Do thf dnh hudng nong dg tde kieh den khd ndng hdp phg dung dich vd do truyin sudl ciia ^- chilm trong 4 gid sir dung thiet hf ddng hda
TAP CHi KHOA HQC & CONG NGHf CAC TRlTdNG DAI HQC KV THUAT * S6 89 - 2012 Thdi gian cQng anh hudng din khi nfing
hip phy ciia chitin (hinh 3). Trong duim djch NaOH 4M, d nhift dO -40''C chitin h ^ phy dung djch eye d^i t^i thdi gian 4h vdi tong lugng hip phy li 1132g trong lODg chitin khd.
ciu triic tinh thi vi lifin kit hydro nOi vi ngo&i trong ciu triic phfin tu p-chilin yiu vi long Ifio hon trong a-chitin nfin P-chitin dl ding hip phy dung djch kiim vi biin ddi ciu triic sang d^ng chitin gel.
Trong sin xuit cdng nghifp, vifc dua nhift do xudng -40°C cd thi lim tfing ding ki chi phf ciia doanh nghifp. Vi v$y, chdng tdi sfi khio sit thfim sy biin dii tfnh chit p-chitin trong mdi trudng kiim vdi sy hd trg cua thiit bj ding hda Phillip J1361. Trong hinh 4, vdi ty If 1:75(g/ml) thyc hifin ddng hda trong thdi gian 40 phiit, d 30°C, thiit bj ddng hda cd tic dyng nhu mdt miy xay tdc dO cao nhim cit mgich chitin va giim kfch thudc hat chitin nfin sin phim sau tic kich truong nd nhifiu han trong
dung djch, cdn d nhift d0 -lO^C ttil qui trinh tic kIch chitin trong mdi trudng kiim vira cit m^h vira phi huy ciu true tinh thi lim cho chitin chuyin sang d^ng gel trong vi bin lim tfing khi ning hip phy eye d^i li 2093g trfin lOOg chitin khd vi dO truyin suit li 71.5%.
3.2 Tfnh chit chitin gel
Chilin gel mSu A: p-chitin phin dng trong mdi trudng kiim NaOH 4M, ty If 1:75(g/ral) d -40''C trong 4 gid tic kfch.
Chilin gel mSu B: p-chitin phin dng trong mdi trudng kiim NaOH 4M, ty Ifi l:75(g/ml) d -lO^C trong 4 gid tic kfch, ding hda 40 phut.
ciu triic bi mit eiia sau khi tic kfch trong mdi trudng kiim d cic difiu kifin cd djnh dugc quan sit dudi kinh hiin vi difin tu qufit SEM. Hinh 5a bfi m^t P-chitin d dang tinh thfi, Hinh 5b, 5c bi m^t mau A vi miu B da chuyfin sang m^ing gel trong sudt.
Hinh 5. (5a) SEM bi mgt cua p-chilin, (5b) SEM be mdt cua chitin gel (mdu A). (5c) SEM be mdt cua chilin gel su dung thiet bi ddng hda (mdu B). Dg phdng dai 1.000. I2kW. lOfim).
Hinh 6. (6a) FTIR0-chitin, (6b) FTIR ehitin gel(mdu A). (6c) FTIR chitin gel(mdu B).
Ill
TAP CHf KHOA HQC A CONG NGHf: CAC TRirdNG DAI HQC KV THUAT * S 6 89 • 2012 Phi FTIR thi hifn cfic nhdm chiirc cd
trong ciu triic P-chitin vi sau khi chuyin sang chitin gel cho thiy tifin kit hydro trong ciu iriic p-chitin di ding bj cit diil dudi tic dyng cCia kiim d nhift dO &m siu (hinh 6). Phi hing ngo^i ciia miu A vi mSu B vSn xuit hifn mQi peak d$c trung ciia nhdm hydroxyl (O-H) vi tifin kit N-H d Wing budc sdng tir 3500 - 3200 cm"'. Phi FTIR cda p-chitin cho thiy c6 lifin kit N-Hdbudc sdng 3431 cm'[9].
Sau khi tuong tic trong dung djch kiim tifin kit niy thay ddi Ifin vimg budc sdng cao hon (3436 cm''d miu A vi 3446 cm'' d mSu B).
Vung budc sdng 1659 - 1625 cm ' trong p- chitin xuit hifn mQi peak ddi d^c trung cua nhdm amide (C^O). nhung sau khi tuong tic trong dung djch kifim thay ddi thinh peak dan (1642 cm"'d mSu A vi 1639 cm"' d mfiu B). Tir cic kit qui cua phi hing ngo?! niy cd thi khing djnh lifn kit hydro n$i vi ngo^i trong ciu tnic p-ehilin dfi bj cit ddt [9].
Mdi trudng kifim vi nhifl dO im sfiu thu$n Igi cho vifc cit dut lifin kfit hydro vi phi hiiy ciu triic tinh thi cua p-chitin. Sau qui trinh tic kich d$ acetyl vi trgng lugng phfin tii ciia chitin gel dfiu glim so vdi p-chitin ban diu (bangl).
Bang 1 DA(%) vd A/„ (Dalton) cda p-chilin.
chilin gel mdu A vd mdu B Thong so
DA(%) M..<Dallon)
P-chilin 85 5.1x10'
Mau A 83 5 5.0x10'
MauB 83 S.OxlO' Phd nhifiu x^ XRD ciia P-chitin cd d^ng peak biu trdn vi cudng dp nhilu x^i m^nh tuong ung cic m§t 010(26 = 8°) vi 110(26 -- 19.5") (hinh 7).
Hinh 7. (7a) Phd nhiiu xg XRD ciia p-chitin.
(7b) XRD cua chitin ^el(mdu A). (7c) XRD ciia chilin gel su dgng ihiii bi ddng hda(m3u B).
Sy phi hiiy ciu irOc tinh the cua p-chilln trong mdi trudng kiim hinh thinh chitin gel vd djnh hinh dfi tim biin ddi hinh d^ng vi gde nhilu X9 ciia phd XRD. Phd XRD ciia miu A vi miu B chi li vfch nhilu khdng xic djnh rd peak (hinh 7b. 7c).
4. K £ T LUJJLN
Khi nfing hip phy vi biin ddi tfnh chit ciia p-chitin trone mdi tnrdng kifim inh hudng nhiiu bdi cic dieu kifn tic kich nhir ndng d$
dung djch kifim, ty If. nhiet d$, vi thjri gtan.
Trong mdi trudng kifim p-chitin hip phy dung djch tiic thdi vi sau 4 gid d^t tr^ng diil cin bing. Kit qua nghifin cuu dfi cho thiy ring khi nfing hip phy dung djch cua chitin ting khi nhift dp giim ho^c ndng d$ kifim tfing. Thifit bj ding hda cd tfic dyng tfing khi nfing hip phy dung djch ciia p-chitin. Sy biin ddi tfnh chit cua chilin dugc xic djnh thdng qua phd nhifiu
\9 XRD, phd hdng ngo^i FTIR vi kfnh hien vi difn tli qufit SEM. Ciu tnic tinh thi ciia pKihitin bj phi hiiy vi t^o thinh chitin gel va tiip phy dung djeh nhifiu nhit d nhiet dd -40''C vdi dung djch NaOH 4M. l> If 1:75 g/ml trong thdi gian 4 gid.
TAI LIf.U THAM KHAO
1. Marguerite Rinaudo. "Chitin and chitosan: Properties and applications". Prog. Polym. Sci, 31 (2006) 603-632.
2. Gardner. K.H. Blackwell, J.. "Refinement of structure of p-chilin", Biopolymers, 14(1975) 1581- 1595.
3 Tolaimate, A.. Desbrieres, J.. Rhazi. M., Alagui. A.. Vincendom. M., Votteri. P., "On the influence of the deacelylation process on thenphysiochemical characteristics of chitosan from squid chitin".
Polymers, 41 (2000) 2463-2469.
TAP CHl KHOA HQC & C6NG NGHp CAC TRirClNC DAI HQC KV TH UAT * S 6 89 - 2012 4. Meriina A. Delorino and Salvacion P. Cresidio, "Investigation of chitosan from Squids pen as Scar
Remover", Worid Applies Sciences Journal, 5 (2009) 98-103.
5. Yongle Liu, Zhaofeng Liu, Wantlan Pan, Qingji Wu, "Absorption behaviors and structure changes of chitin in alkali solution", Carbohydrate Polymers, 72 (2008) 235-239.
6. Focher, B., Beltrame, P.L., Naggi, A., Torri. C , "Alkaline N - deacelylation of ehitin enhanced by flash treatment. Reaction kinetics and structure modification", Carbohydrate Polymers, 12 (1900) 405^18.
7. Rodrigo L. Lavall, Odilio B.G. Assis, Se"rgio P. Campana-Filho, "p-Chitin from the pens of Loligo sp. Extraction and characterization", Bioresource Technology, 98 (2007) 2465-2472.
8. Poirier, M., & Charlet, G. "Chilin fractionation and characterization in N, N- dimethylaeetamide/lithium chloride solvent system"'. Carbohydrate Polymers, 50 (2002) 363-370.
9. Muzzarelli. R. A. A. Natural chelating polymers. New York: Pei^amon Press, (1973) 120-124.
Dia chi lien hi: Ting Thanh Danh - Tel: (+84) 933833681 - Email: [email protected] Khoa KJ Thu^t Hda Hgc. Trudng D?i Hpc Bach khoa Tp.HCM
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