TAP CHl SINH HQC, 2012 34(4) 521 528 NGHIEN ClTU VA iTNG DVNG CONG NGHE M 6 I THAN THI$N M 6 I TRUING K H 6 N G SU* DVNG HOA CHAT TACH CHltj CHITIN TU* VO T 6 M Nguyen v a n Thilt ''''*, Nguyin Ngpc Luon[.]
TAP CHl SINH HQC, 2012 34(4): 521-528 NGHIEN ClTU VA iTNG DVNG CONG NGHE M I THAN THI$N M I TRUING K H N G SU* DVNG HOA CHAT TACH CHltj CHITIN TU* VO T M Nguyen v a n Thilt'*, Nguyin Ngpc L u o n g ^ Trdn Thj Qu^ M a i ^ Hoa Thj HSng', Nguyin Xuan Thy', Nguyin Ngpc Phong'' 'Vifn Cong nghf sinh hpc, *nvlhietibt@yahoo.com ^Vifn Chdn nufli ^Sd Y l l Bdc Giang ""Vifn Khoa hpc vdt lifu TOM TAT: Ldn ddu lien d Vift Nam chiing l6i da nghien ciru irng dyng thdnh cflng cong nghf difn hfla de tach chiet va tinh chi chilin lu vfl ddu tflm, Day 1^ ni^l cong nghf mdi tdch chilt chitin, khong sit dung kiem va acid nhu cac phumig phap khdc, cho nen rdt than thifn vdi moi trudng Qua Irinh difn phan tach chiet chitin dupc thyc hifn ircn ihiet bj mfl hinh kich thudc x 10 x 16,7 cm (dung lich lit) d cac ndng dfl NaCI khac thdi gian 90 phut, Sau difn phdn tach chitin Iren thilt bi nay, dung dich catolite cfl gid trj pH cao nhdl Id 12,43 d nflng dfl NaCI 4%, dung dich anoltle cfl pH thap nhat b^ng 1,9S d ndng dfl NaCI 1% Tu kit qud gid trj pH ciia cac dung djch difn cue va xac dinh ham luong protein dugc chilt rut qua trinh difn phan, dd xac djnh dupc ndng dfl NaCI va thdi gian tfli uu cho qua trinh difn phan tach chiet chitin Ircn thiet bi la 1% va l-l.S h d mSt dfl dong mflt chieu la 400 A/m^ Chi phdm chitin nh^n dupc bang cflng nghf difn hfla cfl dfl sach cao, mdu trSng hojlc trdng hcri vang khong cfl miii la, hdu nhu khong chiia protein, vdi du lupng khoang Ihap (< 0,4%) va khdi lupng phan tu nhdt trung binh td 240 din 1000 kDa Tit khoa: Anolite catolite, chitin, cac chdt khoang, difn phan, protein, vfl torn MO DAU Chitin d u a c phdt hifn vdo ndm 1811, Id mflt polymer sinh hpc n h i l u thd v l lupng ly nhifn, cung voi ddn xudt quan trpng nhdt ciia no la chilosan cfl nhilu tiem ndng dng dung Uong cac linh vyc khdc [3, 4, 5, 9] Trong sudt hai Ihl ky qua va cd hien nay, chitin dupe tdch chilt td cac ngudn nguyfn lifu khac chii ylu bdng p h u a n g phdp hfla hpc, sd dyng xdt vd acid gay fl n h i l m moi d u d n g [ I , 10, 13] Cdc phucmg phdp cflng nghf sinh hpc cdng dd dupc nghien ciht, it gay fl nhilm mfli truflng han, nhung thyc hifn phdc tgp, mdt nhilu thdi gian vd gid enzyme ddt, nfn cQng mfli chi dupe thu nghifm d quy mo pilot [ , 1 , 12, 14] Vift Nam cfl mflt lupng khflng 16 phy phdm vfl ddu tflm rdt gidu protein va chda chilin, mpt polymer sinh hpc cfl rdt nhifu ttiln vpng dng dung cdc ITnh vyc khdc ciia ddi sflng Tuy nhien, ngudn phy phdm ndy dupc chf bien chu y l u bdng phucmg phdp kilm-acid, mpt cdng nghe gdy n h i l m mfli u u d n g vd khflng thu hfli dupc protein co nguyen lifu ban ddu Cdc ca sd s a c h i vd ddu tflm d m i i n Nam Vift Nam sd dyng acid cQng da gdy nhieu fl nhilm cho mfli trudng [16] Vi vdy, vifc nghien cim ung dyng cdc cflng nghe lien tiln de chf bifn ngufln phy phdm qu^ gia ndy cdc sdn pham co gid tri la mflt van de rat cdn thiet Y tudng sd dyng phuang phdp difn hoa tach chill chilin Idn ddu tifn dupc cdc nhd khoa Nga dua id nhung nam 1980 [6] Ban chdt cua phuong phap mdi Id ho^it hoa difn hfla dung dich ciia cdc chdt difn Ii nhu NaCI, kit qiia la tao cac dung djch catolite vd anolite vfli cdc tinh chdt kiim va acid, co tdc dyng thay t h i cho cdc dung dich NaOH va HCl, tuang img, d l loai protein vd cdc chdt khodng khfli vd tflm [6, 7] Han t h i nda, vifc t?io thdnh cdc gflc ty do, cac chdt oxy hda vd cdc chat khu cdc khoang anod vd catod cfln Idm cho chitin dupc tdy mau qud trinh difn phan, kft qua Id khflng cdn phai tien hdnh budc tdy mdu sdn phdm chitin nhdn dupe va quy trinh tdch chilt chitin bdng phucmg phap difn hfla chi gdm cac budc loai protein va loai khodng Ngoai chitin ra, cdng nghf cdn cho phep thu hdi thdnh phan protein cho cdc muc Nguyen Van Thiet et dich dinh duflng khdc Trong cdng trinh ndy chi trinh bdy cdc kit qud nghifin cdu dng dyng cdng nghf mdi ndy dl tdch chilt chitin td vo tflm VAT Lifu \ A PHUONG PHAP NC.H|£N CUU \ (it lifu Nguyen liv'u su dyng cdc thf nghifm tdi uu hfla diL-u kifn tdch chilt chitin Id vfl tflm dd dupc rira s^ch phdn thjt bdm trfn vo, phai khfl vd nghiin (cdt) thdnh cdc mdnh nhd kich thudc 3-5 mm Mudi dn tinh khilt NaCI ddng Idm chdt difn Ii ciia Vift Nam, NaOM, nairi acetate vd acid acetic bdng cua Trung Qudc Thilt bi difn phSn Thilt bi difn phfin phye vy cho myc dich tdch chilt chitin td vfl tflm chung tfli ty thilt kl vd chi t90 (hinh I), gdm binh phan dng A (dung ileh chung Id lit, vdi kich thudc Id X 10 X 16,7 cm), ngudn difn mflt chilu B vd ddy dh\ loyi I (C) vd lo^i II (dung djch cdc khoang difn eye) Binh phdn img A cdu t^o tu khoang: khoang anod (1) vd khoang catod (2) vdi cdc difn eye tuang dng (3 va 4), chung dupc dp sdt vdo cdc thdnh bfn cua binh phan dng; cde khoang difn eye dupc ngdn cdch bdi mflt mdng bdn thdm chpn Ipc cation (5) d chinh gida, ngan ddi binh phdn dng, Thdnh (6) vd ddy (7) binh phdn dng 1dm td mf-ca sudt Hinh Anh thiet bi difn hda cho tdch chiet chitin Nguyen li hoat dpng ctia thiet bj difn hfla Khi cho ddng difn mpt chilu ch^y qua dung dich chat difn Ii, tren anod xay qud trinh oxy hfla nude (2H2O - 4e -> 4H* + O2), cdn U-fn catod-khu nude (2H2O + 2e -* H2 + 20H") Mflt nhdng biln ddi quan trpng nhdt xay cdc khoang difn eye Id sy biln ddi gid Ui pH: khoang catod pH tdng ddn va cfl thi d9t gid trj pH > 12, tuang ducmg vdi pH dung dich kiem m^nh, cfln khoang anod pH biln doi theo chilu ngupc I^i - gidm ddn vd cfl thi dat gid tri pH < [7] Phuvng phdp nghien cuu Ngdm vfl tflm khfl (20 gam) 15 phut dung dich NaCI (theo ti If khfli lupng/thl tich bdng 1:20), sau dfl chuyen vdo khoang catod dl tiln hanh difn phdn Mdi chu trinh tdch chilt gflm idn difn phfin (DPI vd DPII)- Ddu lien nguyen lifu dupc xu li Uong khoang catod df lo^i protein, sau dfl dupc xu Ii khoang anod df lo^ii khodng va tdy mau Sau mfli Idn difn phfin, hfln hpp khoang catod dupc dun ndng 30 phut d 80 ± S^C Sau Idn DPI, sdn phdm dupe tdch khfli dung djch bdng g?ui hay Ipc, sau dfl dupc dua trd l^i cdc khoang tuong img cho Idn DPII Trong qud trinh difn phan, mau cdc dung djch difn eye dupc idy djnh ki 5-10 phut I Idn dl pH vd xdc djnh hdm lupng protein Gid trj pH cua cdc dung djch difn eye dupc tren mdy pH HI 2211 pH/ORP Meter cua hdng Hanna Du lirpng khodngtt-ongchi phdm chitin dupc xdc djnh bdng phuong phdp cdn khdi lupng tro cfln l^i sau khodng hfla bdng ddt mdu Id ddt d nhift dfl cao [15]; lupng protein cfln I?! chi phdm chitin dupe xdc TAP CHI SINH HQC, 2012, 34(4): 521-528 dinh bdng dun mflt lupng mdu nhdt dinh vfli dung dich KOH 2% h vdi ti If chdt khfl/KOH la 1:20 (khdi lupng/thl tfch) d 90'C [2] Ham lupng protein tdt cd cdc mdu dupc xdc djnh bdng phuong phdp quang phd theo hdp thy d 280 nm [8] Khdi lupng phan tu ciia chitin dupc xdc dinh bang phucmg phdp dp nhflt sau dd chuyin tlidnh chilosan bdng ngdm dung dich NaOH 60% ngdy d nhift dfl phdng [15] K £ T QUA VA THAO LU^N Da lien hdnh nghifn cOru tdi im hfla qud trinh difn phdn theo thflng sd quan trpng nhdt: ndng dfl chdt difn 11 NaCI vd thdi gian difn phdn Trong thilt b| dupc sd dyng (hinh 1), cdc difn eye cdch mflt khodng cdch c6 dinh Id 10 cm, cdn mflt thflng sfl khdc Id m$t dfl dflng dien mpt chilu, chung tfli sd dyng gid trj I = 400 A/m^, dya tren thflng bdo ve dflng tdi uu cua cdc 13 Bpr pH • zz:2 ã- t7c c3a ô c6a DPn ãpH 11 tdc gid Nga Id 300-450 A/m^ [7], qud trinh difn phdn tdch chilt chitin dupe tiln hdnh 90 phut d nhift dfl phflng, Kit qud khdo sdt sy biln ddi tfnh chdt kilmaeid qud trinh difn phdn d nhihig ndng dfl NaCI khdc cua ede dung djch difn eye Do myc dfch chfnh ciia qud trinh difn phdn Id 190 dung djch catolite vdi phan Cmg Idlm dl lo^i protein vd anolite vfli phdn dng acid dl lo^i khodng, vi vdy pH cua cdc dung djch difn eye Id thflng sfl quan trpng nhdl cdn dupc khdo sdt Sy biln ddi pH cua dung dich difn eye qud trinh difn phan di dupc khao sdt d cdc ndng dfl NaCI 0,5, 1, 1,5, 2, 3, vd 5% (dupc ki hifu td C| din C7, tuang dng) Kit qud khdo sdt sy biln dfli gia tri pH cdc khoang difn cue t^i cdc thdi diim khdc DPI vd DPII dupc dupc trinh bdy d dgng dd thj trfn hinh • « c2c cSt , ^^^ ^ irz::: — cZa —•—cZe —»t—c5c —8—cla —•—c4a —»t—c7fl ••—-clc »* c4c — • c7c ã c3a ôc7a ã ^ -•—1—•:—'—1 —*—c5a rppMC= •4 ZT.J!„ H—^——1—•— • C3c —•—c6c —8—c2a —.—1—1—.—i- t,min Hinh Dfl thj bilu difn dpng thdi cua pH cde khoang difn eye qud trinh dien phdn Ki hieu tu C] din C7 Id ndng dfl NaCI; o va c chi khoang anod vd catod tuang dng Td cdc dd thi trfn hinh cd thf nit xu hudng biln dfli gid tri pH cdc khoang difn eye d eac ndng dp mudi khac nhu sau: (1) Nhin chung gia tri pH khoang catod tdng lifn tyc, va ngupc lai, gia tri pH khoang anod giam lien tyc qua trinh difn phdn, trd mflt vdi ngoai If; (2) Gid trj pH cdc khoang dien cue biln ddi rdt nhanh 5-10 phut ddu tien cua qud trinh difn phdn, ddc bift DPII, sau biln dfli rdt chdm qud trinh difn phdn tiep theo: sau 30 phut difn phdn gid tti pH khoang catod tdng them khflng nhilu vd gid ttj pH ttong khoang anod gidm di ciing khflng nhilu NIU ddnh gid kha ndng tao dung dich catolite vdi pH cang cao cdng tflt, sg cfl day ndng dfl chdt difn Ii sau: Cj < C| < C4 < C3 < C7 < cs < Cft ddi vdi DPI, va ddy c, < C7 < C4 < cs Nguyen Van Thiet el al < C2 < cj < C6 ddi vdi DPII Tuong ty, cfl cdc ddy ndng dfl NaCI sau ddi vdi kha ndng tgo dung djch anolite vdi pH cdng Uidp cdng tflt: ci < C6< cj < Cj < Cl < Ci < ci dfli vdi DP I vd Cj < Cl < C7 < c^ < Cfi < C3 < C2 ddi vdi DP II Vdi thilt bj difn phdn mfl td ttfn ddy, pH cua dung djch catolite dyt gia trj cao nhdt Id 12,43 ttong DPI vd 12,36 ttong DPII diu d ndng dfl NaCI 4%, cfln dung djch anolite dgt cdc gid tri pH thdp nhdl ttong idn difn phdn tuong dng Id 2,24 vd 1.95 diu d ndng dO NaCI bdng 1% Nhu vdy, so bfl cfl thi kit ludn rdng, trfn thilt bi cua chung tfli, dl tgo dung dich catolite vdi pH cao nhdt, dung djch NaCI 4% Id thich hpp hon cd, cdn dl I90 dung dich anolite cd gid tri pH thdp nhdt, dung dich NaCI 1% Id tdt nhdt Xdc djnh ndng dO NaCI tdi uu vd thdi gian cdn thilt dl xir If vfl tflm khoang catod Trong phuang phdp difn hda tdch chilt chilin, qud ttinh logi protein xdy ttong klioang catod vdi phdn img kiim, cfln qud ttinh 1091 khodng xdy khoang anod vdi phan dng acid VI v$y, gid trj pH ttong cdc khoang catod vd anod Id thudc khd ndng logi bfl protein bdi dung djch catolite vd kha ndng logi cdc chdt khodng bdi dung dich anolite Hinh bilu diln sy phy thuflc ciia pH cdc khoang difn eye tyi cdc thdi diim 30, 60 va 90 phut vdo ndng dfl NaCI \A -^ pH c 30 mm - C, 60 C 90 mm A, 30 - A , 60 A, 90 DP II C, 30 - C , 60 • C, 90 A, 30 —A, 60 A 90 (NaQj, % Hinh Dfl thj bifu difn mfli phy thudc ciia pH ttong cdc khoang difn eye dupc tai cac thdi difm 30, 60 va 90 phut cua qua trinh difn phdn vdo ndng dfl NaCI ttong Idn difn phdn Ki hifu C va A chi cdc dung dich catolite vd anolite, tuang dng Cdc dfl thj trfn hinh cho thdy, gid ttj pH ciia cdc dung dich catolite vd anolite dupc tgi cdc thdi diem 30, 60 vd 90 phiit khdc bift khflng nhifu vd it phy thuflc vdo ndng dO chdt dien ii pH khoang catod sau 30 phdt difn phdn dat gia tti 11,01-12,06; sau 60 phiit dgt gid tti 11,52-12,23 vd sau 90 phut dgt gid tri HJO12,43 Trong dfl, pH khoang anod tgi cdc thdi di6m dgt cde gid tti tuong dng Id: 2,54-3,16; 2,34-3,50 vd 1,95^,00 Han nda, nhin chung gid tri pH ciia dung dich catolite dgt dupc DPII cao han ttong DPI, ngupc Igi, gia tti pH CLia dung djch anolite dgt dupe ttong DPII Igi thdp ban so vdi DPI Nhu vdy, td phdn tich mdi phu thuflc cua gid ttj pH cdc khoang difn cue tgi cac thdi diim 30, 60 vd 90 phut vdo ndng dfl NaCI, cd Ihl kit ludn sa bO dupc rdng sau 30, 60 vd 90 phut difn phdn cac dung djch NaCI nflng dp 0,5-5,0% khoang catod dd trd thdnh dung dich cfl tinh kiim rdt mgnh, cfl kha ndng thay thi dung dich NaOH ho$c KOH vdn dupc sd dyng ttong cdc phuong phdp hda hpc de logi bo protein qud trinh tach chift vd Idm sgeh chitin Tuang ty, eac dung djch anolite tuang dng ciing co tinh acid du mgnh dl logi khoang thay cho dung djch HCl Tuy nhien, cdc kit qua khdo sdt sy biln dfli gid ttj pH ttong cde khoang difn cue vdn chua cho phep xdc djnh mflt cdch cy thf ndng dp tdi uu ciia chdt difn li vd thdi gian cdn phdi tifln hanh qud trinh difn phdn Vi vdy, ddng thai vfli vifc gia ttj pH ttong cde khoang difn cue, da TAP CHI SINH HQC, 2012, 34(4): 521-528 xac djnh cd hdm lupng protein chill nil dupe cde khoang difn eye d cd idn difn phan, Kft qud cde thf nghifm dupe ttlnh bdy dudi dgng dfl thj d hinh fNaCll,% Hinh Dfl thj bilu difn mdi phy thuflc cua hdm lupng protein chilt rut dupc cde khoang difn eye vdo ndng dfl chdt difn li sau 90 phdt difn phdn (A) vd vdo thdi gian difn phdn (doi vdi dung dich catolite) d nflng dfl NaCI 1% (B) Cac dfl thj tren hinh 4A cho thdy, ttong DPI protein dupe chilt nil chu yeu bfli dung dich catolite; tdng nflng dp NaCI lu 0,5% len 5% lupng protein chilt rut dupc bdi dung dich catolite ddu tien tdng, sau dd I^i gidm vd dat nhilu nhdt d ndng dp NaCI 1%, ttong lupng protein chill rut dupc bdi dung djch anolite Igi tdng lien tyc, m^c du il hon nhifu so vfli lupng proteiil dupc chilt nil bdi dung djch catolite Trong DPII lupng protein chilt nit dupc bdi cac dung dich catolite vd anolite gdn tucmg duong nhau, dat nhilu nhdt d ndng dfl NaCI 3%, vd Cling it han nhilu so vfli lupng protein chiflt nit bfli dung djch catolite ttong DPI vd tflm khd ttong khoang catod Id h vd ndng dfl chdt difn Ii NaCI tfli uu cho qud ttinh difn phdn Id 1%, Difn phdn Uong thdi gian I h d nflng dp NaCI 1% cho phep tgo dung dich catolite cfl pH ~I2, tuang ducmg vdi pH dung djch kiim mgnh, thay ihl dupc cho NaOH dl loai protein khfli vfl tom, va dung dich anolite vfli pH ~2, cd tinh acid du manh dl logi khoang Quy trinh difn hfla tach chiet chitin Tu cde kit qud nghifn cdu tim dilu kifn tfli uu cho quy trinh difn hoa tdch chift chitin tu vfl tdm ttinh bay d trfn, chung tfli dd dua quy trinh thu nhdn chi pham chitin bdng phuang phdp difn hoa gdm budc nhu sau: Chudn bi Mat khdc cac dd thi tten hinh 4B lai cho nguyen lifu, Idn dien phan, Idn dun ndng thdy, ham lupng protein dung dich catolite dung dich catolite din 85°C va rua sdn pham tdng lifn tyc ttong 60 phut ddu cua DPI vd ttong bang nude may Mfli chu trinh tdch chift chilin 50 phut ddu cua DPII, sau hdu nhu khflng gdm Idn difn phdn Trong chu trinh difn phan thay ddi tdng tilp thdi gian difn phdn ddu tien chi cd khoang catod chda nguyen lifu cd DPI vd DPII Dilu ndy cd nghTa Id dl logi bfl vd tflm Td chu trinh dien phdn thu ttd di ca protein ciia vfl tflm kho bdng phuang phdp difn khoang difn cue diu chda nguyfn lifu: khoang hfla, cdn dien phan it nhdt 60 phiit cdc catod chda me vd tflm mfli, khoang anod chira dilu kifn da mfl ta d tren ban thdnh phdm dd dupe loai protein td khoang Nhu vdy, tir kit qua khao sal sy biln ddi gid catod cua chu trinh dien phdn trudc, d day nfl tti pH vd xdc dinh ham lupng protein ttong cdc dupc logi khodng va lay mau, sg cho sdn phdm chitin sach Chf pham chitin tdch chilt bang dung djch difn eye tgi cde thdi difm khdc cflng nghf mfli cd dfl sgch cao, mau trdng ttong qua trinh difn phdn ttinh bay d tten ed the dep ho^c trdng hoi vang (hinh 5) thdy rdng, thfli gian cdn thilt (tSi im) cho xd Ii 525 Nguyen Van Thiet et al •^•^^^v* Hinh Chi phdm chitin tdch Lhiei bang phuang phdp difn hfla chyp trfn cdc ndn khdc Cdc chi phdm chitin tdch chili bdng phuang phdp difn hfla hdu nhu khflng chda protein vd chi chiia mdt hdm lupng nhfl cdc chdt khodng (< 0,4% ttxi), cfl khdi l u ^ g phfin tu nhdl trung binh Mv = 240-1000 kDa, phy thuflc vdo mdu vfl tflm Ktl LUAN Td cdc kit qud nghifn cim ttinh bdy tten day cfl thi rut cdc ket ludn sau: Dd thilt kl vd chi tgo dupc mflt thilt bj mfl hinh df nghien cdu vd dp dyng phuang phdp difn hda tdch chilt chitin td vd tom Thilt bi ndy cho phfp tgo dung djch catolite vdi pH 12,0-12,43 vd dung djch anod vdi pH 1,95-2,5, CO the su dyng thay Ihl cho kiim (NaOH) vd acid (HCl) dl logi protein vd cdc chdt khodng tuang ung, ttong cdc quy trinh tdch chiet va tinh chf chitin td vd tflm Da xac dinh dupc cdc dieu kifn tdi uu cho quy trinh difn phdn tdch chilt chitin td vfl tom ttfn thilt bj mfl hinh ndy Id [NaCI] = 1,0%, thdi gian difn phdn t = 1-1,5 h d mdt dd ddng difn mflt chilu khdng dfli Id 400 A/m^ vd ti If vfl tflm khfl/dung djch chdt difn Ii Id 1:20 (khdi lupng/thl tich) Dd dua quy trinh difn hfla tdch chilt chitin td vo ddu tom gdm budc^ Chudn bj nguyen lifu, Idn difn phan, idn dun dung dich catolite d 85 ± 5°C 30 phut vd rua san pham bdng nude mdy din pH trung tinh Chf pham chitin nhdn dupc bdng cflng nghf mdi ndy cfl mdu ttdng ho^c ttdng hai vdng, khong cfl mui vj Ig, chda -0,4% khodng, hdu nhu khdng cd chda protein, cd khdi lupng phdn td nhdt td 240-1000 kDa, phy thuflc vdo mlu nguyen Hfu 526 L&i c&m an: Cflng ttinh dupc sy hS trp kinh phi ciia dl tdi "Nghifn cdu dng dyng enzyme tgo nanochitin dl sdn xudt biosorbent su dyn^ cflng nghifp dupc" (2011-2012) thuflc Dfl dn phdt triln vd img dyng cflng nghf sinh hpc ITnh vyc cdng nghifp chi biln din ndm 2020 Bfl Cflng Thuang qudn li TAI LI$U THAM KHAO Bykova V.M., Nemlsep S.V., 2002 Raw material sources and methods for producing chitin and chilosan, pp.7-23 In: Skryabin K G., Vikhorevaya G A., Variamov V P (Eds), 2002 Chitin and Chilosan: Production, Properties and Usage, Publishing house "Nauka", 368 p (in Russian) Cremades 0., Ponce E., Corpas R et al., 2001 Processing of crawfish (Procamparus clarkii) for the preparation of carotenoproteirm and chitin J Agric Food Chem., 49(11); 5468-5472 Felse P A Panda T., 1999 Smdies on applications of chitin and its derivatives Bioprocess Eng., 20: 505-512 Nguyin Thj Huf, Ldm Ngpc Thy, Nguyfn Qudc Hien, Nguyfn Vdn Hoan, 2001 Nghien cim tde dyng eiia cdc chdt cfl hogt tfnh sinh hpc cao tu chitin ddi vdi sy mdm cua hgt thflc gidng Tap ehi Hod hpc, 39(3): 23-26 Knorr D., 1991 Recovery and utilisation of chitin and chitosans in food processing waste management Food Technol., 45; 114122 Kuprina N E., Vodolazhskaya S., 2002 TAP CHI SINH HQC, 2012,34(4): 521-528 Methods of isolation and activation of chitin and chilosan, pp 44-63 In; Skryabin K G., Vikhorevaya G A., Variamov V P (Eds), 2002 Chitin and Chilosan: Production, Properties and Usage, Publishing house "Nauka", 368 p (in Russian) Maslova G V., 2002 Theory and practice for chitin isolation by electtochemical method, pp 24-43 In: Skryabin K G., Vikhorevaya G A., Variamov V P (Eds), 2002 Chitin and Chilosan: Production, Properties and Usage, Publishing house "Nauka", 368 p (in Russian) Meshkova N P and Severin S E (eds.), 1979 Handbook on Biochemistry, published by Moscow Univ Press, 430 p (in Russian) Suzuki S., 2001 Recent advances in Biomedical Application of chitie substances, in Chilin and Chilosan in Life Science, Eds Uragami T Kurita K., and Fukamizo T., Kodansha Scientific Ltd., Japan, 2001 10 Nguyfn Vdn Thilt, 2006 Xdy dung quy ttinh cflng nghf tflng hpp chdt co lac dung chdng vifm khdp tu vfl tom Bdo cao tdng kit dl tdi cdp Bp - Vifn Khoa hpc vd Cflng nghe Viet Nam 2004-2005 11 Nguyin van Thiet DS Ngpc Tu, Nguyin Thanh Hdi, 2005 Mflt sd kit qud budc ddu nghien cdu quy trinh san xudt chitin td vfl tflm bdng phuang phdp cflng nghf enzyme Tuyin tdp cdc bdo cdo khoa hpc tgi Hfli nghi Mfli ttudng todn qudc 2005, Hd Nfli: 14281432 12 Nguyin Vdn Thilt, Do Ngpc Tu, 2007 Nghien ciru tdch chilt chitin tir ddu-vfl tflm bang cdc phuang phdp sinh hpc I - Sd dyng bromelain ttong djch fp vfl dda Tgp chi Klioa hpc vd Cong nghf, 45(3): 51-58, 13 Mai Xudn Tjnh, 2001 Dilu chi chitin/ chilosan td vfl tflm phi thdi Tap chi Hod hpc vd Cong nghf hdu ca, 4(69): 17-19 14 Dfl Ngpc Tu Nguyin Vdn Thilt, 2006 Phuang phdp Ifn men vfli chiing vi khudn sinh proteinasse tdch chiet chitin td vd tflm Tgp chi Dupc lifu, 11(3): 132-136 15 Variamov V P., Nemtsev S V-., Tikhonov V E (eds.), 2010 Chitin and Chilosan: Nature, Production and Applications Materials of Project CYTED TV 14: Chitin and Chilosan from Crustacean Processing Wastes Translated from Spanish by K M: Mikhlina, E V Zhukova, E S Krylova, Published by the Russian Chilin Society, 292 p (in Russian) RESEARCH AND APPLICATION OF NEW ENVIROMENTAL-FRENDLY TECHNOLOGY WITHOUT USING CHEMICALS FOR EXTRACTION OF CHITIN FROM SHRIM SHELLS Nguyen Van Thiet'*, Nguyen Ngoc Luong\ Tran Thi Quy Mai^ Hoa Thi Hang', Nguyen Xuan Thu', Nguyen Ngoc Phong" 'institute of Biotechnology, VAST ^National Instimte of Animal Science ^Bac Giang Department of Health ^Institute of Materials Science, VAST Viettiam has a huge amount ofrich-proteinshrimp shells containing chitin, a biological polymer that has many potential applications in different areas of life However, this shrimp by-products are processed mainly by alkali-acid method, causing environmental pollution and can not recovery the protein in the starting material For the first time in Vietnam, we have carried out research and successfiilly applied electtochemical Nguyen Van Thiet et al technology to extract and purify the chitin from shrimp shells This Is a new technology for chilin extraction, that Mul use ulkiili and iicid as in the other chciiiiciil methods, so it is very friendly to the environment The electrolysis is pcrfonned on the model device of size ofd x 10 X 16,7 cm (with a capacity of liter) at different NaCI conccntrulions in 90 minutes After electrolytic extraction of chilin on the device the highest pH value of catolite solution i.s 12.43 at a concentration of 4% NaCI, and the lowest pH of anolile is 1.95 al NaCI concentrations of 1% From the results of measuring the pH value of die electrode solutions and dcicmiination of protein extracted in (he electrode cells were determined the optimal electrolyte concentration (that is I % of NaCI) and minimal time (thiil is 1.0- hours) in DC cun-cnt of 400 A/m^ Chitin preparations obtained by electtochemical technology have high purity, while or yellowish-white colour, no foreign odors, virtually no protein, with low mineral residues (< 0,4%) (ind average viscosimctric molecular weight from 240 to 1000 kDa, Keywords: Anolite, catolite, chitin, electrolysis, mincruls, protein, shrim shells Ngdy nhdn bdi: 20-5-2012 ... tach chiet chitin Tu cde kit qud nghifn cdu tim dilu kifn tfli uu cho quy trinh difn hoa tdch chift chitin tu vfl tdm ttinh bay d trfn, chung tfli dd dua quy trinh thu nhdn chi pham chitin bdng... khodngtt-ongchi phdm chitin dupc xdc djnh bdng phuong phdp cdn khdi lupng tro cfln l^i sau khodng hfla bdng ddt mdu Id ddt d nhift dfl cao [15]; lupng protein cfln I?! chi phdm chitin dupe xdc TAP... nghf mdi ndy dl tdch chilt chitin td vo tflm VAT Lifu \ A PHUONG PHAP NC.H|£N CUU \ (it lifu Nguyen liv''u su dyng cdc thf nghifm tdi uu hfla diL-u kifn tdch chilt chitin Id vfl tflm dd dupc rira