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Reactive & Functional Polymers 46 (2000) 127 www.elsevier.

com/locate/react

Review

review o! c"itin an# c"itosan a$$lications


&a'eti (.). Ravi *umar %
+e$artment o! ,"emistry- .niversity o! Roor/ee- Roor/ee 247 667- 0n#ia Receive# 24 1anuary 20002 receive# in revise# !orm 20 1une 20002 acce$te# 23 1une 2000

4stract ,"itin is t"e most a4un#ant natural amino $olysacc"ari#e an# is estimate# to 4e $ro#uce# annually almost as muc" as cellulose. 0t "as 4ecome o! 5reat interest not only as an un#erutili6e# resource- 4ut also as a new !unctional material o! "i5" $otential in various 7el#s- an# recent $ro5ress in c"itin c"emistry is 8uite notewort"y. 9"e $ur$ose o! t"is review is to ta/e a closer loo/ at c"itin an# c"itosan a$$lications. :ase# on current researc" an# e;istin5 $ro#ucts- some new an# !uturistic 2000 <lsevier =cience :.). ll ri5"ts reserve#. a$$roac"es in t"is !ascinatin5 area are t"orou5"ly #iscusse#.
*eywor#s> :ea#s2 :iotec"nolo5y2 ,"itin2 ,"itosan2 ,ontrolle# #ru5 #elivery2 Fi4ers2 (ano$articles2 ?y#ro5els2 9a4lets2 9rans#ermal #evices

1. 0ntro#uction ,"itin- a naturally a4un#ant muco$olysac@ c"ari#e- an# t"e su$$ortin5 material o! crusta@ ceans- insects- etc.- is well /nown to consist o! 2@acetami#o@2@#eo;y@b @+@5lucose t"rou5" a b (1 4) lin/a5e. ,"itin can 4e #e5ra#e# 4y c"itinase. 0ts immuno5enicity is e;ce$tionally low- in s$ite o! t"e $resence o! nitro5en. 0t is a "i5"ly insolu4le material resem4lin5 cellulose in its solu4ility an# low c"emical reactivity. 0t may 4e re5ar#e# as cellulose wit" "y#ro;yl at
q 9"is $a$er is #e#icate# to Pro!essor &.(.). Prasa#- P".+.F(0< ((ew +el"i)- +=c. ("c ,olom4o)- =c"ool o! Ai!e =ciences.niversity o! ?y#era4a#- ?y#era4a#- 0n#ia- w"o ins$ire# me wit" "is scienti7c a$$roac"- "onesty an# "uman warmt". %Post :a5 (o. 2E- Roor/ee 247 667- 0n#ia. Fa;> 1 E1@1GG2@ 7G360. <@mail a##ress> mnvr/umarHmailcity.com (&.(.). Ravi *umar).

$osition ,@2 re$lace# 4y an acetami#o 5rou$. Ai/e cellulose- it !unctions naturally as a struc@ tural $olysacc"ari#e. ,"itin is a w"ite- "ar#inelastic- nitro5enous $olysacc"ari#e an# t"e ma'or source o! sur!ace $ollution in coastal areas. ,"itosan is t"e (@#eacetylate# #erivative o! c"itin- alt"ou5" t"is (@#eacetylation is al@ most never com$lete. s"ar$ nomenclature wit" res$ect to t"e #e5ree o! (@#eacetylation "as not 4een #e7ne# 4etween c"itin an# c"itosan B1-2C. 9"e structures o! cellulose- c"itin an# c"itosan are s"own in Fi5. 1. ,"itin an# c"itosan are o! commercial interest #ue to t"eir "i5" $ercenta5e o! nitro5en (6.DEF) com$are# to synt"etically su4stitute# cellulose (1.23F). 9"is ma/es c"itin a use!ul c"elatin5 a5ent B1C. s most o! t"e $resent@#ay $olymers are syn@ t"etic materials- t"eir 4iocom$ati4ility an# 4io#e5ra#a4ility are muc" more limite# t"an t"ose o! natural $olymers suc" as cellulose-

1GD1@314D /00/I see !ront matter P00> =1GD1@314D(00)000GD@E

2000 <lsevier =cience :.). ll ri5"ts reserve#.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

ra#a4ility- non@to;icity- a#sor$tion $ro$ertiesetc. Recently- muc" attention "as 4een $ai# to c"itosan as a $otential $olysacc"ari#e resource B3C. lt"ou5" several e!!orts "ave 4een re$orte# to $re$are !unctional #erivatives o! c"itosan 4y c"emical mo#i7cations B6DC- very !ew attaine# solu4ility in 5eneral or5anic solvents BE-10C an# some 4inary solvent systems B111GC. ,"emi@ cally mo#i7e# c"itin an# c"itosan structures resultin5 in im$rove# solu4ility in 5eneral or@ 5anic solvents "ave 4een re$orte# 4y many wor/ers B142GC. 9"e $resent review is an attem$t to #iscuss t"e current a$$lications an# !uture $ros$ects o! c"itin an# c"itosan.

2. Processin5 o! c"itin an# c"itosan ,"itin is easily o4taine# !rom cra4 or s"rim$ s"ells an# !un5al mycelia. 0n t"e 7rst casec"itin $ro#uction is associate# wit" !oo# in#us@ tries suc" as s"rim$ cannin5. 0n t"e secon# caset"e $ro#uction o! c"itosan5lucan com$le;es is associate# wit" !ermentation $rocesses- similar to t"ose !or t"e $ro#uction o! citric aci# !rom s$er5illus ni5er- &ucor rou;ii- an# =tre$@ tomyces- w"ic" involves al/ali treatment yiel#@ in5 c"itosan5lucan com$le;es. 9"e al/ali re@ moves t"e $rotein an# #eacetylates c"itin simul@ taneously. +e$en#in5 on t"e al/ali concentra@

Fi5. 1. =tructures o! cellulose- c"itin an# c"itosan.

c"itin- c"itosan an# t"eir #erivatives. ?owevert"ese naturally a4un#ant materials also e;"i4it a limitation in t"eir reactivity an# $rocessa4ility BG-4C. 0n t"is res$ect- c"itin an# c"itosan are recommen#e# as suita4le !unctional materials4ecause t"ese natural $olymers "ave e;cellent $ro$erties suc" as 4iocom$ati4ility- 4io#e5@

=c"eme 1.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

tion- some solu4le 5lycans are remove# B24C. 9"e $rocessin5 o! crustacean s"ells mainly involves t"e removal o! $roteins an# t"e #isso@ lution o! calcium car4onate w"ic" is $resent in cra4 s"ells in "i5" concentrations. 9"e resultin5 c"itin is #eacetylate# in 40F so#ium "y#ro;i#e at 120 8, !or 1G ". 9"is treatment $ro#uces 70F #eacetylate# c"itosan (=c"eme 1). G. <conomic as$ects 9"e $ro#uction o! c"itin an# c"itosan is currently 4ase# on cra4 an# s"rim$ s"ells #iscar#e# 4y t"e cannin5 in#ustries in Jre5onLas"in5ton- )ir5inia an# 1a$an an# 4y various 7nis"in5 Keets in t"e ntarctic. =everal coun@ tries $ossess lar5e une;$loite# crustacean re@ sources- e.5. (orway- &e;ico an# ,"ile B23C. 9"e $ro#uction o! c"itosan !rom crustacean s"ells o4taine# as a !oo# in#ustry waste is economically !easi4le- es$ecially i! it inclu#es t"e recovery o! carotenoi#s. 9"e s"ells contain consi#era4le 8uantities o! asta;ant"in- a carot@ enoi# t"at "as so !ar not 4een synt"esi6e#- an# w"ic" is mar/ete# as a 7s" !oo# a##itive in a8uaculture- es$ecially !or salmon. 9o $ro#uce 1 /5 o! 70F #eacetylate# c"itosan !rom s"rim$ s"ells- 6.G /5 o! ?,l an# 1.D /5 o! (aJ? are re8uire# in a##ition to nitro5en- $rocess water (0.3 t) an# coolin5 water (0.E t). 0m$ortant items !or estimatin5 t"e $ro#uction cost inclu#e trans$ortation- w"ic" varies #e$en#in5 on la4or an# location. 0n 0n#iat"e ,entral 0nstitute o! Fis"eries 9ec"nolo5y*erala- initiate# researc" on c"itin an# c"itosan. From t"eir investi5ation- t"ey !oun# t"at #ry $rawn waste containe# 2GF an# #ry s8uilla containe# 13F c"itin B26C. 9"ey "ave also re$orte# t"at t"e c"itinous soli# waste !raction o! t"e avera5e 0n#ian lan#in5 o! s"ell 7s" ran5es !rom 60 000 to D0 000 tonnes B27-2DC. ,"itin an# c"itosan are now $ro#uce# commer@ cially in 0n#ia- 1a$an- Polan#- (orway an# ustralia. 9"e worl#wi#e $rice o! c"itosan (in small 8uantities) is ca. .=I7.3/10 5 (=i5ma an# l#ric" $rice list).

4. Pro$erties o! c"itin an# c"itosan &ost o! t"e naturally occurrin5 $olysac@ c"ari#es- e.5. cellulose- #e;tran- $ectin- al5inic aci#- a5ar- a5arose an# carra5enans- are neutral or aci#ic in nature- w"ereas c"itin an# c"itosan are e;am$les o! "i5"ly 4asic $olysacc"ari#es. 9"eir uni8ue $ro$erties inclu#e $olyo;ysalt !ormation- a4ility to !orm 7lms- c"elate metal ions an# o$tical structural c"aracteristics B2EC. Ai/e cellulose- c"itin !unctions naturally as a structural $olysacc"ari#e- 4ut #i!!ers !rom cellu@ lose in its $ro$erties. ,"itin is "i5"ly "y#ro@ $"o4ic an# is insolu4le in water an# most or5anic solvents. 0t is solu4le in "e;aKuoro@ iso$ro$anol- "e;aKuoroacetone- c"loroalco"ols in con'u5ation wit" a8ueous solutions o! miner@ al aci#s B24C an# #imet"ylacetami#e containin5 3F lit"ium c"lori#e. ,"itosan- t"e #eacetylate# $ro#uct o! c"itin- is solu4le in #ilute aci#s suc" as acetic aci#- !ormic aci#- etc. Recently- t"e 5el !ormin5 a4ility o! c"itosan in (@met"ylmor@ $"oline (@o;i#e an# its a$$lication in controlle# #ru5 release !ormulations "as 4een re$orte# BG0G2C. 9"e "y#rolysis o! c"itin wit" concen@ trate# aci#s un#er #rastic con#itions $ro#uces relatively $ure +@5lucosamine. 9"e nitro5en content o! c"itin varies !rom 3 to DF #e$en#in5 on t"e e;tent o! #eacetylationw"ereas t"e nitro5en in c"itosan is mostly in t"e !orm o! $rimary ali$"atic amino 5rou$s. ,"itosan- t"ere!ore- un#er5oes reactions ty$ical o! amines- o! w"ic" (@acylation an# =c"i!! reaction are t"e most im$ortant. ,"itosan #e@ rivatives are easily o4taine# un#er mil# con@ #itions an# can 4e consi#ere# as su4stitute# 5lucans. (@ cylation wit" aci# an"y#ri#es or acyl "ali#es intro#uces ami#o 5rou$s at t"e c"itosan nitro5en. cetic an"y#ri#e a!!or#s !ully acetylate# c"itins. Ainear ali$"atic (@acyl 5rou$s a4ove $ro$ionyl $ermit ra$i# acetylation o! "y#ro;yl 5rou$s. ?i5"er 4en6oylate# c"itin is solu4le in 4en6yl alco"ol- #imet"ylsul!o;i#e!ormic aci# an# #ic"loroacetic aci#. 9"e (@ "e;anoyl- (@#ecanoyl an# (@#o#ecanoyl #eriva@

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

tives "ave 4een o4taine# in met"anesul!onic aci# BGG-G4C. t room tem$erature- c"itosan !orms al@ #imines an# /etimines wit" al#e"y#es an# /etones- res$ectively. Reaction wit" /etoaci#s !ollowe# 4y reaction wit" so#ium 4oro"y#ri#e $ro#uces 5lucans carryin5 $roteic an# non@ $roteic amino 5rou$s. (@,ar4o;ymet"yl c"itosan is o4taine# !rom 5lyo;ylic aci#. <;am@ $les o! non@$roteic amine aci# 5lucans #erive# !rom c"itosan are t"e (@car4o;y4en6yl c"itosans o4taine# !rom o@ an# $@$"t"alal@ #e"y#ic aci#s B24-23C. ,"itosan an# sim$le al#e"y#es $ro#uce (@al/yl c"itosan u$on "y#ro@ 5enation. 9"e $resence o! t"e more or less 4ul/y su4stituent wea/ens t"e "y#ro5en 4on#s o! c"itosan2 t"ere!ore (@al/yl c"itosans swell in water in s$ite o! t"e "y#ro$"o4icity o! t"e al/yl c"ains- 4ut t"ey retain t"e 7lm !ormin5 $ro$erty o! c"itosan B1C.

t"e universally acce$te# non@to;ic (@#e@ acetylate# #erivative o! c"itin- w"ere c"itin is (@#eacetylate# to suc" an e;tent t"at it 4ecomes solu4le in #ilute a8ueous acetic an# !ormic aci#s. 0n c"itin- t"e acetylate# units $revail (#e5ree o! acetylation ty$ically 0.E0). ,"itosan is t"e !ully or $artially (@#eacetylate# #erivative o! c"itin wit" a ty$ical #e5ree o! acetylation o! less t"an 0.G3. 9o #e7ne t"is ratio- attem$ts "ave 4een ma#e wit" many analytical tools BG344C- w"ic" inclu#e 0R s$ectrosco$y$yrolysis 5as c"romato5ra$"y- 5el $ermeation c"romato5ra$"y an# .) s$ectro$"otometry1 7rst #erivative o! .) s$ectro$"otometry?@ 1G (&R s$ectrosco$y- , soli# state (&R- t"er@ mal analysis- various titration sc"emes- aci# "y#rolysis an# ?PA,- se$aration s$ectrometry met"o#s an#- more recently- near@in!rare# s$ec@ trosco$y B43C.

4.1. P"ysical an# c"emical c"aracteri6ation


9"e structural #etails o! cellulose- c"itin an# c"itosan are s"own in Fi5. 1. ,ellulose is a "omo$olymer- w"ile c"itin an# c"itosan are "etero$olymers. (eit"er ran#om nor 4loc/ orientation is meant to 4e im$lie# !or c"itin an# c"itosan. 9"e $ro$erties o! c"itin an# c"itosan suc" as t"e ori5in o! t"e material (#iscusse# in t"e $revious section)- t"e #e5ree o! (@#eacetyla@ tion- molecular wei5"t an# solvent an# solution $ro$erties are #iscusse# in 4rie!. Mlycol c"itin- a $artially J@"y#ro;yet"ylate# c"itin- was t"e 7rst #erivative o! $ractical im$ortance2 ot"er #erivatives an# t"eir $ro$ose# uses are s"own in 9a4le 1.

4.1.2. &olecular wei5"t ,"itosan molecular wei5"t #istri4utions "ave 4een o4taine# usin5 ?PA, B46C. 9"e wei5"t@ avera5e molecular wei5"t (& ) w o! c"itin an# c"itosan "as 4een #etermine# 4y li5"t scatterin5 B47C. )iscometry is a sim$le an# ra$i# met"o# !or t"e #etermination o! molecular wei5"t2 t"e constants a an# * in t"e &ar/?ouwin/ e8uation "ave 4een #etermine# in 0.1 & acetic aci# an# 0.2 & so#ium c"lori#e solution. 9"e intrinsic viscosity is e;$resse# as
Bh C 5 *&a 5 1.D1 3 10 2 &
G 0.EG

4.1.1. +e5ree o! (@acetylation n im$ortant $arameter to e;amine closely is t"e #e5ree o! (@acetylation in c"itin- i.e. t"e ratio o! 2@acetami#o@2@#eo;y@+@5luco$yranose to 2@amino@2@#eo;y@+@5luco$yranose structural units. 9"is ratio "as a stri/in5 e!!ect on c"itin solu4ility an# solution $ro$erties. ,"itosan is

9"e c"ar5e# nature o! c"itosan in aci# solvents an# c"itosanNs $ro$ensity to !orm a55re5ation com$le;es re8uire care w"en a$$lyin5 t"ese constants. Furt"ermore- convertin5 c"itin into c"itosan lowers t"e molecular wei5"t- c"an5es t"e #e5ree o! #eacetylation- an# t"ere4y alters t"e c"ar5e #istri4ution- w"ic" in turn inKuences t"e a55lomeration. 9"e wei5"t@avera5e molecu@ 6 6 lar wei5"t o! c"itin is 1.0G 3 10 to 2.3 3 10 4ut t"e (@#eacetylation reaction re#uces t"is to 3 3 1 3 10 to 3 3 10 B4DC.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 9a4le 1 ,"itin #erivatives an# t"eir $ro$ose# uses +erivative (@ cyl c"itosans <;am$les Formyl- acetyl- $ro$ionyl- 4utyryl- "e;anoyloctanoyl- #ecanoyl- #o#ecanoyl- tetra#ecanoyllauroyl- myristoyl- $almitoyl- stearoyl- 4en6oylmonoc"loroacetoyl- #ic"loroacetyl- triKuoroacetylcar4amoyl- succinyl- aceto;y4en6oyl (@,ar4o;y4en6yl- 5lycine@5lucan ((@car4o;y@ met"yl c"itosan)- alanine 5lucan- $"enylalanine 5lucan- tyrosine 5lucan- serine 5lucan- 5lutamic aci# 5lucan- met"ionine 5lucan- leucine 5lucan From an"y#ri#es suc" as maleic- itaconic- acetyl@ t"iosuccinic- 5lutaric- cyclo"e;ane 1-2@#icar4o;@ ylic- $"t"alic- cis@tetra"y#ro$"t"alic- 3@nor4o@ rnene@2-G@#icar4o;ylic- #i$"enic- salicylic- tri@ mellitic- $yromellitic an"y#ri#e o@,ar4o;ymet"yl- crosslin/e# o@car4o;ymet"yl Potential uses 9e;tiles- mem4ranes an# me#ical ai#s

(@,ar4o;yal/yl (aryl) c"itosans

,"romato5ra$"ic me#ia an# metal ion collection O

(@,ar4o;yacyl c"itosans

o@,ar4o;yal/yl c"itosans

&olecular sievesviscosity 4uil#ersan# metal ion collec@ tion O

=u5ar #erivatives

1@+eo;y5alactic@1@yl@- 1@#eo;y5lucit@1@yl@1@#eo;ymeli4iit@1@yl@- 1@#eo;ylactit@1@yl@1@#eo;ylactit@1@yl@4(2-2-6-6@tetramet"yl$i$eri#ine@ @1@o;yl)@- 1@#eo;y@6 9@al#e"y#olactit@1@yl@1@#eo;y@69@al#e"y#omeli4iit@1@yl@- cello4iit@1@yl@ c"itosans- $ro#ucts o4taine# !rom ascor4ic aci# Palla#ium- co$$er- silver- io#ine

&etal ion c"elates

,atalyst- $"oto5ra$"y"ealt" $ro#ucts- an# insectici#es 9e;tiles

=emisynt"etic resins o! c"itosan (atural $olysacc"ar@ i#e com$le;esmiscellaneous

,o$olymer o! c"itosan wit" met"yl met"acrylate$olyurea@uret"ane- $oly(ami#eester)- acrylami#e@ maleic an"y#ri#e ,"itosan 5lucans !rom various or5anisms l/yl c"itin- 4en6yl c"itin ?y#ro;y 4utyl c"itin- cyanoet"yl c"itosan

?y#ro;y et"yl 5lycol c"itosan Mlutaral#e"y#e c"itosan Ainoelic aci#c"itosan com$le; .racylc"itosan- t"eo$"ylline c"itosan- a#enine@ c"itosan- c"itosan salts o! aci# $olysacc"ari#esc"itosan stre$tomycin- 2@ami#o@2-6@#iamino"e$@ tanoic aci# c"itosan

Flocculation an# metal ion c"elation 0nterme#iate- serine $rotease $uri7cation +esaltin5 7ltration#ialysis an# insulatin5 $a$ers <n6ymolo5y- #ialysis an# s$ecial $a$ers <n6yme immo4ili6ation Foo# a##itive an# antic"olesterolemic

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

4.1.G. =olvent an# solution $ro$erties :ot" cellulose an# c"itin are "i5"ly crys@ talline- intracta4le materials an# only a limite# num4er o! solvents are /nown w"ic" are a$$lic@ a4le as reaction solvents. ,"itin an# c"itosan #e5ra#e 4e!ore meltin5- w"ic" is ty$ical !or $olysacc"ari#es wit" e;tensive "y#ro5en 4on#@ in5. 9"is ma/es it necessary to #issolve c"itin an# c"itosan in an a$$ro$riate solvent system to im$art !unctionality. For eac" solvent system$olymer concentration- $?- counterion concen@ tration an# tem$erature e!!ects on t"e solution viscosity must 4e /nown. ,om$arative #ata !rom solvent to solvent are not availa4le. s a 5eneral rule- t"e ma;imum amount o! $olymer is #issolve# in a 5iven solvent towar#s a "omo5eneous solution. =u4se8uently- t"e $oly@ mer is re5enerate# in t"e re8uire# !orm (#is@ cusse# in t"e !ollowin5 sections). coa5ulant is re8uire# !or $olymer re5eneration or soli#i7ca@ tion. 9"e nature o! t"e coa5ulant is also "i5"ly #e$en#ent on t"e solvent an# solution $ro$erties as well as t"e $olymer use# B34-73C.
3. ,"itin an# its #erivatives in 74re !ormation

3.1. (atural micro74riller arran5ement


,"itin "as 4een /nown to !orm micro74rillar arran5ements in livin5 or5anisms. 9"ese 74rils are usually em4e##e# in a $rotein matri; an# "ave #iameters !rom 2.3 to 2.D nm. ,rustacean cuticles $ossess c"itin micro74rils wit" #iame@ ters as lar5e as 23 nm. 9"e $resence o! mi@ cro74rils su55ests t"at c"itin "as c"aracteristics w"ic" ma/e it a 5oo# can#i#ate !or 74re s$innin5. 9o s$in c"itin or c"itosan 74res- t"e raw $olymer must 4e suita4ly re#issolve# a!ter removal o! e;traneous material suc" as calcium car4onate an# $roteins- w"ic" encase t"e mi@ cro74rils.

3.2. Fi4re !ormation R in retros$ection


(umerous met"o#s o! s$innin5 c"itin 74res "ave 4een re$orte# since )on Leimarn re$orte#

t"e 7rst solutions o! c"itin t"at coul# 4e !orme# into a Pro$y@$lasticN state in 1E26. ?e $re$are# t"e solution usin5 inor5anic salts ca$a4le o! stron5 "y#ration B4EC- suc" as Ai,(=,a(,(=) 2 - ,a0 -2,a:r - ,a,l - etc. !ter t"is 2 2 re$ort- many solvent systems inclu#in5 or5anic solvents an# mi;tures o! inor5anic salts an# or5anic solvents came into e;istence. 9o "el$ t"e #issolution o! c"itin- it was (@ #eacetylate# in 3F caustic so#a at 60 8, !or 14 #ays B30C. not"er $roce#ure !or (@#eacetyla@ tion was to $lace t"e c"itin in an autoclave !or G " at 1D0 8, an# 10 atm $ressure. 0t was $ointe# out t"at 6 to 10F o! soli#s o! (@#eacetylate# c"itin can 4e 4rou5"t into aci#ic solution at room tem$erature. 8ueous acetic aci# was !oun# to 4e suita4le !or t"is $ur$ose. !ter $assin5 t"e $olymer solutions t"rou5" a 7lter $ress to remove im$urities- 74res were s$un. ,"emicals incom$ati4le wit" c"itin were su55este# as coa5ulants. 9"e resultant 74res were was"e# an# #rie# un#er tension. 9"e 7nal $ro#uct 74res "a# a roun#@ to "eart@s"a$e# cross section wit" a tensile 4rea/in5 loa# o! G3 2 /5/mm (G43 Pa). 9"e 74res $ossesse# a #ull luster similar to natural sil/- lea#in5 to t"e su55estion t"at t"e (@#eacetylate# c"itin 74res woul# ma/e 5oo# arti7cial "air. 9"e collection an# recyclin5 o! c"itin !rom small@scale con@ sumers was also su55este#. ,lar/ an# =mit" re$orte# a $roce#ure !or $ro#ucin5 74res 4y #issolution o! c"itin at E3 8, in $resaturate# solutions o! lit"ium t"iocyanate (saturate# 60 8,) B31C. (o tensile $ro$erties or solution concen@ trations were re$orte#. ?owever- Q@ray analysis s"owe# a "i5" #e5ree o! orientation. =olvent removal was not success!ul even at 200 8,. Ait"ium io#i#e was im$lie# to "ave 4e"ave# in t"e same manner. ratio o! 3 mol lit"ium t"iocyanate $er mole an"y#ro5lucose unit was !oun# to e;ist. 9"is is com$ara4le to t"e celluloselit"ium t"iocyanate com$oun#. ,ellu@ lose solu4ility an# t"e role o! solvate/salt com$le;es "ave 4een reviewe# in #etail B32-3GC. Recently- Rat"/e an# ?u#son $u4lis"e# a re@ view "i5"li5"tin5 t"e a4ility o! c"itin an# c"itosan as 74re an# 7lm !ormers B34C.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

3.G. (ovel solvent s$in systems 3.G.1. ?alo5enate# solvent s$in system 0n 1E73- ustin su55este# or5anic solvents containin5 aci#s !or t"e #irect #issolution o! c"itin. =uc" a system was c"loroet"anol an# sul!uric aci#. 9"e $reci$itation o! c"itin in 74rillar !orm in water- met"anol- or a8ueous ammonium "y#ro;i#e was mentione#- 4ut no 74re tensile #ata were $resente# B33C. 0n 1E73- :rine an# ustin su55este# tri@ c"loroacetic aci# (9, ) as a c"itin solvent. ,"itin was $ulveri6e# an# two $arts 4y wei5"t were a##e# to D7 $arts 4y wei5"t o! a solvent mi;ture containin5 40F 9, - 40F c"loral "y#rate (.= +e$artment o! 1ustice- +ru5 <n@ !orcement 5ency- class 0) controlle# su4@ stance)- an# 20F met"ylene c"lori#e over a $erio# o! G043 min. 7lament was e;tru#e# !rom t"is solution usin5 a "y$o#ermic nee#le an# acetone as t"e coa5ulant. 9"e 7lament was t"en neutrali6e# wit" $otassium "y#ro;i#e (*J?) in 2@$ro$anol !ollowe# 4y was"in5 in #eioni6e# water. 9"e 7laments were t"en col# #rawn. 9wo tensile 4rea/s were ta/en at 60F relative "umi#ity an# room tem$erature. 9"e 7rst was !rom a 7lament wit" a cross section o! 0.0D 3 0.10 mm- yiel#in5 a tensile stren5t" o! 72 2 /5/mm (710 Pa) an# a 4rea/in5 elon5ation o! 1GF. 9"e secon# 7lament "a# a cross section o! 0.014 3 0.740 mm- in#icatin5 a colla$se# core structure. 0t "a# a tensile stren5t" o! 104 /5/ 2 mm (1026 Pa) an# a 4rea/in5 elon5ation o! 44F B36C. =yrin5in5 a 7lament cannot 4e inter$rete# as conclusive evi#ence !or a $ossi4le wet s$innin5 $rocess. L"ile syrin5e e;trusion mi5"t in#icate t"e selection o! a coa5ulant- it woul# 4e rat"er sur$risin5 to o4tain meanin5!ul tensile #ata. ="ear !orces in a s$inneret are muc" 5reater t"an t"ose e;$erience# in a sy@ rin5e ti$. *i!une an# co@wor/ers su55este# #issolvin5 c"itin in 9, an# a c"lorinate# "y#rocar4on suc" as c"loromet"ane- #ic"loromet"ane- an# 1-1-2@tric"loroet"ane. 9"e 9, concentration s"oul# 4e /e$t 4etween 23 an# 73F. con@ centration ran5e 4etween 1 an# 10F c"itin was

su55este# as well as #issolution 4elow room tem$erature. Fi4res were e;tru#e# t"rou5" a s$inneret o! 0.04 an# 0.06 mm #iameter into an acetone coa5ulation 4at" !ollowe# 4y a met"a@ nol 4at". 9"e tensile stren5t" o! #rie# 7laments was in t"e ran5e o! 1.67 to G.1 5/# wit" an elon5ation !rom D.7 to 20.0F. 9"e stren5t" o! t"e 74res was im$rove# 4y leavin5 t"em in a 0.3 5/l a8ueous caustic so#a solution !or 1 ". 9"e resultant tensile stren5t"s were 2.23 to G.20 5/# wit" elon5ations o! 1E.2 to 27.GF- res$ec@ tively B37C. *i!une an# co@wor/ers !urt"er su5@ 5este# t"at t"ese c"itin 7laments were suita4le as a4sor4a4le sur5ical suture B3DC. ?owever9, is very corrosive an# #e5ra#es t"e $oly@ mer molecular wei5"t. 9"e 4rea/in5 elon5ations su55est t"at t"e "alo5enate# solvents act as $lastici6ers. Fu'i =$innin5 ,om$any #issolve# c"itosan in a mi;ture o! water an# #ic"loroacetic aci# (+, ). 9"e 6.44F c"itosan acetate salt solution viscosity was 410 $oise. 9"e #o$e was e;tru#e# t"rou5" a $latinum no66le (G0 "oles o! 0.2 mm #iameter eac") into 4asic ,u,J ((? )J?4 G solution to !orm 74res. +enier an# tensile $ro$erties were not re$orte# B3EC. 9o/ura an# co@wor/ers use# a com4ination o! !ormic aci# (F )- +, an# #iiso$ro$yl et"er as a solvent system. ,"itin was cycle# several times !rom 2 208, to room tem$erature in F !ollowe# 4y a##ition o! a small amount o! +, . +iiso$ro$yl et"er was t"en a##e# to re#uce t"e solution viscosity to 4elow 1EE $oise an# tensile $ro$erties were also re$orte# B60C. 0t is notewort"y t"at t"e wet stren5t" #ro$s to 4elow 0.30 5/# 4ut t"at t"e elon5ation increases to 1GF. 9, /#ic"loromet"ane s$in system is also #escri4e# 4y t"e .niti/a ,o. 9"ree $arts c"itin were #issolve# in 30 $arts 9, an# 30 $arts #ic"loromet"ane. 9"e #e!oame# #o$e was e;@ tru#e# into acetone 4e!ore win#@u$. 9"e 4o4@ 4ins were neutrali6e# wit" *J?- was"e# wit" water- an# #rie#. 9"e 74res "a# a tensile stren5t" o! 2 5/# an# 0.320 #enier B61C. .niti/a ,o. also use# t"e 9, /c"loral "y@ #rate/#ic"loroet"ane solvent system !or c"itin.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

Five $arts were #issolve# in 100 $arts o! a 4>4>2 9, /c"loral "y#rate/#ic"loroet"ane solvent mi;ture an# e;tru#e# t"rou5" a 0.06 mm no66le into acetone. 9"e 74res were treate# wit" met"anolic (aJ?. 9"e o$timum 74res 5ave a tenacity o! G.2 5/# wit" an elon5ation o! 20F B62C. .niti/a ,o. !ollowe# t"is u$ wit" anot"er $atent usin5 a 60>40 9, /tric"loroet"ylene s$in #o$e mi;ture. 9ensile $ro$erties were unavaila4le B6GC. 0n 1EDG- .niti/a ,o. s"owe# t"at a #o$e consistin5 o! t"ree $arts c"itin- 30 $arts 9, - an# 30 $arts #ic"loromet"ane coul# 4e s$un at a rate o! 1.7 ml/min un#er 23 2 /5/cm $ressure into acetone to !orm 7laments. 9"e e;trusion #ie "a# "oles o! 0.07 mm #iam@ eter- in#icatin5 a 'et velocity o! D.D m/min an# a ta/e@u$ o! 3 m/min. 9"e coa5ulation 4at" was maintaine# at 1D 8,. 9"e 7laments were was"e# wit" acetone at 1D 8, !or 10 min- rewoun# at 4.3 m/min- t"en neutrali6e#- was"e# an# #rie#. 9"e multi7lament $ro#uct "a# a total #enier o! 130 wit" a tenacity o! 2.63 5/# B6GC. similar system usin5 !our $arts c"itin in t"e same solvent 4ut a 40@"ole #ie o! 0.0D mm #iameter eac" was also use#. 9"e 'et velocity was 10.4 m/min into a 23 8, acetone 4at". rewin#in5 at 7 m/min !ollowe# t"e 7rst ta/e@u$ roll at 3 m/min. 9"e total #enier was 1732 "owever- no tensile $ro$erties were re$orte# B64C. =ome o! t"e "alo5enate# solvent systems attaine# #ry tenacities o! a4ove G 5/#2 "owevert"e low wet tenacities were still un#esira4le. lt"ou5" t"e 74re c"aracteri6ation was muc" 4etter !or t"ese systems- t"e $olymer c"aracteri@ 6ation lac/e# molecular wei5"t as well as #e5ree o! (@acetylation !ormation. =olution $ro$erties woul# 4e "ar# to o4tain #ue to ra$i# c"itin #e5ra#ation in t"ese solvents. lt"ou5" an"y#rous coa5ulation 4at"s were use# an# com$are#- 74res were neutrali6e# in a8ueous me#ia. stu#y in com$letely an"y#rous sys@ tems woul# 4e o! interest- since it may lea# to more #ensely consoli#ate# 74res. 9"e im@ $lementation o! t"ese s$in systems re$resents a $ro4lem #ue to t"e nature o! t"e solvents. 9, an# +, are corrosive an# #e5ra#e t"e $olymer

u$on s"ort e;$osures. ,"loro"y#rocar4ons are increasin5ly environmentally unacce$ta4le sol@ vents. ?e;aKuoro@2@$ro$anol an# "e;aKuoro@ acetone ses8ui"y#rate are to;ic. Formic aci# can act as a sensiti6er.

3.G.2. mi#eAi,l system 0n 1E7D- Rut"er!or# an# ustin summari6e# t"e $ro4lems encountere# in 7n#in5 a solvent system !or c"itin B63C. ustin su55este# (-(@ #imet"ylacetami#e (+& c)3F Ai,l or (@ met"yl@2@$yrroli#one ((&P)3F Ai,l as sol@ vents !or c"itin. solution o! 3F w/v was o4taine# wit"in 2 " wit" t"ese systems. 7lament was e;tru#e# !rom t"e solution usin5 a 13@5au5e nee#le into an acetone coa5ulation 4at". 9"is was !ollowe# 4y more was"in5 an# #rawin5 in acetone. 9"e 7nal 7lament was was"e# in #eioni6e# water. 9ensile $ro$erties were o4taine# at 60F R.?. an# room tem$era@ ture at an a$$lie# stress o! 0.1 cm/min. 9"e resultant #ry tensile stren5t"s !or #i!!erent cra4 an# s"rim$ s$ecies ran5e# !rom 24 to 60 /5/ 2 mm (2G63E2 Pa) B66C. Russian researc"ers s$un c"itin 74res out o! +& c/(&P solutions containin5 3F c"itin an# 3F Ai,l (4ase# on c"itin content). 9"ese 74res were #rawn in a 30>30 et"anol/et"ylene 5lycol 4at"- 5ivin5 an avera5e yiel# stren5t" o! GE0 &Pa wit" GF elon5ation. n initial mo#ulus o! 2 MPa was also re$orte#. =cannin5 electron microsco$y s"owe# 74res wit" a roun# 74rillar cross section B67C. !ollow@u$ stu#y s"owe# a #ecrease in t"e elasticity mo#ulus an# relative elon5ation wit" increase in t"e #e5ree o! (@acetylation (12G0F). From Q@ray analy@ sis- an increase in t"e amount o! amor$"ous re5ions was o4serve# wit" increase in #e5ree o! acetylation B6DC. 9"e ami#elit"ium systems s"owe# some o! t"e 4est #ry tenacities- alt"ou5" t"ey still lac/ a#e8uate wet tenacities. 9"e low wet tenacities are $ro4a4ly #ue to low crystallinity an# $oor consoli#ation o! t"e 74re. 9"e 74res an# s$in #o$es were well c"aracteri6e# 4ut t"e $olymers use# to $re$are t"ese #o$es were not. =ome

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

coa5ulation stu#ies were carrie# out 4ut a clear com$arison coul# not 4e ma#e. $ro4lem wit" t"is s$in system is t"e removal an# recovery o! lit"ium !rom t"e 74re. 9"e lit"ium acts as a Aewis aci# 4y solvatin5 t"e c"itin ami#e 5rou$. 0t is unclear i! t"is can 4e com$letely reverse# t"rou5" was"in5- once t"e 74res are !orme#.

6.1. P"oto5ra$"y
,"itosan "as im$ortant a$$lications in $"oto@ 5ra$"y #ue to its resistance to a4rasion- its o$tical c"aracteristics- an# 7lm !ormin5 a4ility. =ilver com$le;es are not a$$recia4ly retaine# 4y c"itosan an# t"ere!ore can easily 4e $ene@ trate# !rom one layer to anot"er o! a 7lm 4y #i!!usion B70C.

3.G.G. mine o;i#e/water system ttem$ts "ave 4een ma#e to #evelo$ a $ro@ cess !or c"itosan 74res 4y #irect #issolution usin5 a novel solvent system- (@met"ylmor@ $"oline o;i#e/water ((&&J/? J)-24ut no interestin5 tensile #ata were o4taine# !rom t"ese $reliminary investi5ations B6EC.
6. $$lications

6.2. ,osmetics
For cosmetic a$$lications- or5anic aci#s are usually 5oo# solvents- c"itin an# c"itosan "ave !un5ici#al an# !un5istatic $ro$erties. ,"itosan is t"e only natural cationic 5um t"at 4ecomes viscous on 4ein5 neutrali6e# wit" aci#. 9"ese materials are use# in creams- lotions an# $erma@ nent wavin5 lotions an# several #erivatives "ave also 4een re$orte# as nail lac8uers B7DC.

9"e interest in c"itin ori5inates !rom t"e stu#y o! t"e 4e"aviour an# c"emical c"aracteris@ tics o! lyso6yme- an en6yme $resent in "uman 4o#y Kui#s B70C. wi#e variety o! me#ical a$$lications !or c"itin an# c"itin #erivatives "ave 4een re$orte# over t"e last t"ree #eca#es B717GC. 0t "as 4een su55este# t"at c"itosan may 4e use# to in"i4it 74ro$lasia in woun# "ealin5 an# to $romote tissue 5rowt" an# #i!!erentiation in tissue culture B74C. 9"e $oor solu4ility o! c"itin is t"e ma'or limitin5 !actor in its utili6ation. +es$ite t"is limitation- various a$$lications o! c"itin an# mo#i7e# c"itins "ave 4een re$orte#- e.5. as raw material !or man@ma#e 74res B34C. Fi4res ma#e o! c"itin an# c"itosan are use!ul as a4sor4@ a4le sutures an# woun#@#ressin5 materials B3D-73-76C. ,"itin sutures resist attac/ in 4ileurine an# $ancreatic 'uice- w"ic" are $ro4lem areas wit" ot"er a4sor4a4le sutures B3DC. 0t "as 4een claime# t"at woun# #ressin5s ma#e o! c"itin an# c"itosan 74res "ave a$$lications in wastewater treatment. ?ere- t"e removal o! "eavy metal ions 4y c"itosan t"rou5" c"elation "as receive# muc" attention B70-77C. 9"eir use in t"e a$$aral in#ustry- wit" a muc" lar5er sco$e- coul# 4e a lon5@term $ossi4ility B7DC.

6.G. ,"itosan as an arti7cial s/in


0n#ivi#uals w"o "ave su!!ere# e;tensive loss@ es o! s/in- commonly in 7res- are actually ill an# in #an5er o! succum4in5 eit"er to massive in!ection or to severe Kui# loss. Patients must o!ten co$e wit" $ro4lems o! re"a4ilitation aris@ in5 !rom #ee$- #is75urin5 scars an# cri$$lin5 contractures. &alette et al. stu#ie# t"e e!!ect o! treatment wit" c"itosan an# saline solution on "ealin5 an# 74ro$lasia o! woun#s ma#e 4y scal$el insertions in s/in an# su4cutaneous tissue in t"e a4#ominal sur!ace o! #o5s B7EC. Sannas et al. $ro$ose# a #esi5n !or arti7cial s/in- a$$lica4le to lon5@term c"ronic use- !ocus@ in5 on a nonanti5enic mem4rane- w"ic" $er@ !orms as a 4io#e5ra#a4le tem$late !or synt"esis o! neo#ermal tissue BD0C. 0t a$$ears t"at c"itosan- "avin5 structural c"aracteristics simi@ lar to 5lycosamino 5lycans- coul# 4e consi#ere# !or #evelo$in5 suc" su4stratum !or s/in re$lace@ ment BD1DGC.

6.G.1. ,"itin@ an# c"itosan@4ase# #ressin5s ,"itin an# c"itosan "ave many #istinctive 4iome#ical $ro$erties. ?owever- c"itin@4ase#

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&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

woun# "ealin5 $ro#ucts are still at t"e early sta5es o! researc" BD4C. =$ar/es an# &urray BD3C #evelo$e# a sur@ 5ical #ressin5 ma#e o! a c"itosan5elatin com@ $le;. 9"e $roce#ure involves #issolvin5 t"e c"itosan in water in t"e $resence o! a suita4le aci#- maintainin5 t"e $? o! t"e solution at a4out 2G- !ollowe# 4y a##in5 t"e 5elatin #issolve# in water. 9"e ratio o! c"itosan an# 5elatin is G>1 to 1>G. 9o re#uce t"e sti!!ness o! t"e resultin5 #ressin5 a certain amount o! $lastici6ers suc" as 5lycerol an# sor4itol coul# 4e a##e# to t"e mi;ture. +ressin5 7lm was cast !rom t"is solution on a Kat $late an# #rie# at room tem$erature. 0t was claime# t"at- in contrast to conventional 4iolo5ical #ressin5s- t"is e;@ $erimental #ressin5 #is$laye# e;cellent a#"e@ sion to su4cutaneous !at. (ara et al. BD6C $atente# a woun# #ressin5 com$risin5 a nonwoven !a4ric com$ose# o! c"itin 74res ma#e 4y t"e wet s$innin5 tec"@ ni8ue. 0n one o! t"e e;am$les- c"itin $ow#er was 5roun# to 100 mes" an# treate# in 1 & ?,l !or 1 " at 4 8,. 0t was t"en "eate# to E0 8, w"ere it was treate# !or G " in a 0.GF (aJ? solution to remove calcium an# $rotein in t"e c"itin $ow#er- an# rinse# re$eate#ly !ollowe# 4y #ryin5. 9"e resultant c"itin was #issolve# in a #imet"ylacetami#e solution containin5 7 wtF lit"ium c"lori#e to !orm a 7F #o$e. !ter 7lterin5 an# allowin5 #e!oamin5 to occur- t"e #o$e was e;tru#e# t"rou5" a no66le o! #iameter 0.06 mm an# 200 "oles into 4utanol at 60 8, a t a rate o! 2.2 5/min. 9"e c"itin was coa5ulate# an# collecte# at a s$ee# o! 10 m/min. 9"e resultant stran# was rinse# wit" water an# #rie# to o4tain a 7lament o! 0.74 #te; wit" a stren5t" o! 2.D 5/#en. 9"e 7laments were t"en cut into sta$le 74res. .sin5 $oly(vinyl alco"ol) as a 74rous 4in#er- nonwoven #ressin5s were ma#e. *i!une et al. BD7C #evelo$e# a new woun# #ressin5- :esc"itin L- com$ose# o! c"itin non@ woven !a4ric w"ic" $rove# to 4e 4ene7cial in clinical $ractice. *im an# &in BDDC "ave #e@ velo$e# a woun#@coverin5 material !rom $oly@ electrolyte com$le;es o! c"itosan wit" sul!on@ ate# c"itosan. 0t is $ro$ose# t"at woun# "ealin5

is accelerate# 4y t"e oli5omers o! #e5ra#e# c"itosan 4y tissue en6ymes an# t"is material was !oun# to 4e e!!ective in re5eneratin5 t"e s/in tissue in t"e area o! t"e woun#. :ia5ini et al. BDEC #evelo$e# an (@car4o;y@ 4utyl c"itosan #ressin5 !or treatin5 $lastic sur5ery #onor sites. solution o! (@car4o;y@ 4utyl c"itosan was #ialy6e# an# !ree6e@#rie# to G $ro#uce a 10 3 20 3 0.3 cm so!t an# Ke;i4le $a#- w"ic" was sterili6e# an# a$$lie# to t"e woun#. 9"is #ressin5 coul# $romote or#ere# tissue re5eneration com$are# to control #onor sites. :etter "istoarc"itectural or#er- 4etter vas@ culari6ation an# t"e a4sence o! inKammatory cells were o4serve# at t"e #ermal level- w"ile !ewer as$ects o! $roli!eration o! t"e mal$i5"ian layer were re$orte# at t"e e$i#ermal level. 9"e :ritis" 9e;tile 9ec"nolo5y Mrou$ (:99M) $atente# a $roce#ure !or ma/in5 a c"itin@4ase# 74rous #ressin5s BE0EGC. 0n t"is met"o# t"e c"itin/c"itosan 74res were not ma#e 4y t"e tra#itional 74re@s$innin5 tec"ni8ue an# t"e raw materials were not !rom s"rim$ s"ell 4ut !rom micro@!un5i instea#. 9"e $roce#ure can 4e summari6e# as !ollows. (i) &icro@!un5al mycelia $re$aration !rom a culture o! &ucor muce#o 5rowin5 in a nutrient solution. (ii) ,ulture was"in5 an# treatment wit" (aJ? to remove $rotein an# $reci$itate c"itin/c"itosan. (iii) :leac"in5 an# !urt"er was"in5. (iv) Pre$aration o! t"e #is$ersion o! 74res usin5 $a$er@ma/in5 e8ui$ment. (v) Filtration an# wet@lai# matt $re$aration2 mi;in5 wit" ot"er 74res to 5ive mec"anical stren5t". 9"is is a novel met"o#- w"ic" uses a non@ animal source as t"e raw material- an# t"e resultin5 micro@!un5al 74res are totally #i!!erent !rom normal s$un 74res. 9"ey "ave "i5"ly 4ranc"e# an# irre5ular structures. 9"e 74res are unmana5ea4ly 4rittle w"en t"ey are allowe# to #ry an# a $lastici6er "as to 4e associate# wit" t"e w"ole $rocess an# a wet@lai# matt is use# as t"e 4asic $ro#uct.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

11

Recently- &u66arelli BE4C intro#uce# anot"er c"itosan #erivative3@met"yl$yrroli#inone c"itosan- w"ic" is 4elieve# to 4e very $romisin5 in me#ical a$$lications. 9"is $olymer is claime# to 4e com$ati4le wit" ot"er $olymer solutionsinclu#in5 5elatin- $oly(vinyl alco"ol)- $oly@ (vinyl $yrroli#one) an# "yaluronic aci#. 9"e a#vanta5es inclu#e "ealin5 o! woun#e# mensi@ cal tissues- an# o! #ecu4itus ulcers- #e$ression o! ca$sule !ormation aroun# $rost"eses- limita@ tion o! scar !ormation an# retraction #urin5 "ealin5. =ome woun#@#ressin5 sam$les were $re$are# !rom an a8ueous solution o! t"is 3@ met"yl$yrroli#one c"itosan- w"ic" was #ialy6e# an# laminate# 4etween stainless steel $lates an# !ree6e@#rie# to yiel# Keeces. 9"e material coul# 4e !a4ricate# into many #i!!erent !orms- suc" as 7laments- nonwoven !a4rics- etc. Jnce a$$lie# to a woun#- 3@met"yl$yrroli#inone c"itosan 4ecomes availa4le in t"e !orm o! oli5omers $ro#uce# un#er t"e action o! lyso6yme. not"er c"itin #erivative- #i4utyrylc"itinwas $re$are# 4y treatment o! /rill c"itin wit" 4utyric an"y#ri#e in t"e $resence o! $erc"loric aci# as a catalyst at 23G0 8, BE3C. =am$les o! $olymers wit" molecular wei5"ts "i5" enou5" to !orm 74res were o4taine# an# #i4utyryl c"itin 74res were ma#e 4y #ry s$innin5 a 20 22F solution into acetone. 9"e 74res "ave tensile $ro$erties similar to or 4etter t"an t"ose o! c"itin. &oreover- it was claime# t"at c"itin 74res wit" 5oo# tensile $ro$erties coul# 4e o4taine# 4y al/aline "y#rolysis o! #i4utyryl c"itin 74res wit"out #estroyin5 t"e 74re struc@ ture. s !ar as c"itin@4ase# commercial woun# #ressin5s are concerne#- one $ro#uct (:esc"itin - .niti/a) is commercially availa4le in 1a$an- w"ic" is a nonwoven !a4ric manu!ac@ ture# !rom c"itin 7laments.

!or #i5estion o! mil/ lactose. ,owNs mil/ con@ tains only a limite# amount o! t"e ( M moiety"ence some in!ants !e# cowNs mil/ may "ave in#i5estion. &any animals an# some "umans (inclu#in5 t"e el#erly) "ave similar lactose intolerances BE6-E7C. nimal nutritional stu#ies "ave s"own t"at t"e utili6ation o! w"ey may 4e im$rove# i! t"e #iet contains small amounts o! c"itinous materi@ al. 9"is im$rovement is attri4ute# to t"e c"an5e in t"e intestinal microKora 4rou5"t a4out 4y t"e c"itinous su$$lement BEDC. ,"ic/ens !e# a com@ mercial 4roiler #iet containin5 20F #rie# w"ey an# 2 or 0.3F c"itin "a# si5ni7cantly im$rove# wei5"t a5ain com$are# to controls BEE-100C. 9"e !ee# e!7ciency ratio s"i!te# !rom 2.3 to 2.GD #ue to incor$oration o! c"itin in t"e !ee# B100C.

6.3. J$t"almolo5y
,"itosan $ossesses all t"e c"aracteristics re@ 8uire# !or ma/in5 an i#eal contact lens> o$tical clarity- mec"anical sta4ility- su!7cient o$tical correction- 5as $ermea4ility- $articularly to@ war#s o;y5en- wetta4ility an# immunolo5ical com$ati4ility. ,ontact lenses are ma#e !rom $artially #e$olymeri6e# an# $uri7e# s8ui# $en c"itosan 4y s$in castin5 tec"nolo5y an# t"ese contact lenses are clear- tou5" an# $ossess ot"er re8uire# $"ysical $ro$erties suc" as mo#ulustensile stren5t"- tear stren5t"- elon5ation- water content an# o;y5en $ermea4ility. 9"e anti@ micro4ial an# woun# "ealin5 $ro$erties o! c"itosan alon5 wit" an e;cellent 7lm ca$a4ility ma/e c"itosan suita4le !or #evelo$ment o! ocular 4an#a5e lenses B101C.

6.6. Later en5ineerin5


s environmental $rotection is 4ecomin5 an im$ortant 5lo4al $ro4lem- t"e relevant in#us@ tries $ay attention to t"e #evelo$ment o! tec"@ nolo5y w"ic" #oes not cause environmental $ro4lems.

6.4. Foo# an# nutrition


9"e (@acetyl5lucosamine (( M) moiety $resent in "uman mil/ $romotes t"e 5rowt" o! 4i7#o 4acteria- w"ic" 4loc/ ot"er ty$es o! microor5anism an# 5enerate t"e lactase re8uire#

6.6.1. &etal ca$ture !rom wastewater (air an# &a#"avan B102C use# c"itosan !or

12

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

t"e removal o! mercury !rom solutions- an# t"e a#sor$tion /inetics o! mercuric ions 4y c"itosan were re$orte# 4y Penic"e@covas et al. B10GC. 9"e results in#icate t"at t"e e!7ciency o! a#@ 2 sor$tion o! ?5 1 4y c"itosan #e$en#s u$on t"e $erio# o! treatment- t"e $article si6e- initial 2 concentration o! ?5 1 an# 8uantity o! c"itosan. 1"a et al. B104C stu#ie# t"e a#sor$tion o! 2 ,# 1 on c"itosan $ow#er over t"e concentration ran5e o! 110 $$m usin5 various $article si6es 4y a#o$tin5 a similar $roce#ure as !or t"e removal o! mercury. ?y#ro;ymet"yl c"itin an# ot"er water@solu@ 4le #erivatives are use!ul Kocculents !or anionic waste streams. ,"itosan (@4en6ylsul!onate #e@ rivatives were use# as sor4ents !or removal o! metal ions in an aci#ic me#ium 4y Leltrows/i et al. B103C. 9"e selective a#sor$tion ca$acity !or metal ions o! ami#o;imate# c"itosan 4ea#@ 5@P ( co$olymer "as 4een stu#ie# 4y *an5 et al. B106C. 9"ese investi5ations clearly in#icate t"at c"itosan "as a natural selectivity !or "eavy metal ions an# is use!ul !or t"e treatment o! wastewater. &c*ay et al. B107C use# c"itosan !or t"e 2 2 2 2 removal o! ,u 1 - ?5 1 - (i 1 an# Tn 1 wit"in t"e tem$erature ran5e 2360 8, at near neutral $?. Furt"er a#sor$tion $arameters !or t"e re@ moval o! t"ese metal ions were re$orte# 4y San5 et al. B10DC. &aruca et al. B10EC use# c"itosan Ka/es o! 0.44 mm !or t"e removal o! ,r(000) !rom wastewater. 9"e a#sor$tion ca$aci@ ty increase# wit" a #ecrease in t"e si6e o! t"e Ka/es- w"ic" im$lie# t"at metal ions were $re!era4ly a#sor4e# on t"e outer sur!ace o! c"itosan in t"e removal o! ,r(000) !rom t"e wastewater. Pseu#o@7rst@or#er /inetics are re@ $orte#.

6.6.2. ,olour removal !rom te;tile mill e!Kuents 6.6.2.1. =or$tion o! #yes (o sin5le #ecolori6ation met"o# is li/ely to 4e t"e o$timum !or all wastewater streams

B110C. +ue to its uni8ue molecular structurec"itosan "as an e;tremely "i5" a!7nity !or many classes o! #yes- inclu#in5 #is$erse- #irect- reac@ tive- aci#- vat- sul!ur an# na$"t"ol #yes. 9"e rate o! #i!!usion o! #yes in c"itosan is similar to t"at in cellulose. Jnly !or 4asic #yes "as c"itosan a low a!7nity. ,"itosan is versatile in sor4in5 metals an# sur!actants- as well as to #erivati6ation to attract 4asic #yes an# ot"er moieties (e.5.- $roteins !rom !oo# $rocessin5 $lants). 9"e sor$tion o! #yes 4y c"itosan is e;ot"er@ mic- an increase in t"e tem$erature lea#s to an increase in t"e #ye sor$tion rate- 4ut #iminis"es total sor$tion ca$acity B111C. ?owever- t"ese e!!ects are small an# normal wastewater tem@ $erature variations #o not si5ni7cantly a!!ect t"e overall #ecolori6ation $er!ormance B112C. lsot"e wastewater $? may 4e an im$ortant !actor in t"e sor$tion o! certain #yes onto c"itosan 4ecause- at low $?- c"itosanNs !ree amino 5rou$s are $rotonate#- causin5 t"em to attract anionic #yes. ,ontact time or- inversely- Ku; (wastewater Kow $er unit cross@sectional area) a!!ects sor$tion in a com$le; manner in a 7;e# 4e# #esi5n reactor system #ue to contact time4e# $enetration an# 4oun#ary layer e!!ects. t "i5" Ku;- t"e #iversion o! li8ui# into lar5er c"annels aroun# $articles an# tur4ulent Kow occur. 0n 5eneral- a low Ku; ten#s to 5ive more com$lete contaminant removal. For almost all t"e treatment strate5ies- a ma'or !actor w"ic" "as not yet 4een a#e8uately c"aracteri6e# is t"e e!!ect o! ty$ical wastewater contaminants on #ecolori6ation e!7ciencies. 0n ty$ical #yein5 systems it is well /nown t"at certain a##itives suc" as salt an# sur!actants can eit"er accelerate or retar# #ye sor$tion $ro@ cesses. 9"e e;treme varia4ility o! te;tile waste@ water must 4e ta/en into account in t"e #esi5n o! any #ecolori6ation system. Finally- a !actor w"ic" si5ni7cantly increases t"e sor$tion rate is t"e loa#in5 t"ermo#ynamicsw"ic" in#icates w"et"er a reaction is !avoure#. s loa#in5 increases- t"e #rivin5 !orces !or sor$tion #ecrease- lea#in5 to an ultimate satura@

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

1G

tion value 4eyon# w"ic" !urt"er sor$tion is not $ossi4le.

ran5e 2.07.0 t"e #ye@4in#in5 ca$acity o! c"itin was s"own to 4e sta4le- w"ile c"itosan !orme# 5els 4elow $? 3.3 an# coul# not 4e evaluate#.

6.6.2.2. +ye@4in#in5 $ro$erties o! c"itin an# c"itosan *norr e;amine# t"e #ye@4in#in5 $ro$erties 4y wei5"in5 0.3 or 2.0 5 c"itin or c"itosan in centri!u5e tu4es- a##in5 20 5 o! a8ueous #ye solution (3 to 4E m5 #ye/l) an# t"en s"a/in5 t"e close# centri!u5e tu4es !or G0 min at 200 r$m in a "ori6ontal $osition. 9"e sam$les were t"en centri!u5e# !or G3 min at 4300 3 5- t"e su$ernatant #ecante# an# t"e water u$ta/e o! c"itin an# c"itosan #etermine# a!ter =osuls/i B11GC. 9"e a4sor4ance o! t"e su$ernatant was measure# at 303 nm usin5 #ecolori6e# water as a 4lan/. 9"e wei5"t o! t"e su$ernatant was use# as t"e 4asis !or t"e calculation o! t"e total amount o! #ye 4oun# or release#. $? a#'ust@ ment was carrie# out 4y usin5 eit"er 10 ml o! a commercial 4u!!er solution or 4y a##in5 0.1 & ?,l to a slurry o! 0.3 5 c"itin/c"itosan an# 10 ml o! #ye solution. !ter stirrin5 !or 13 min- t"e $? was rea#'uste# an# #eioni6e# water a##e# to 20.3 5 total wei5"t. ,"itosan !orme# 5els at $? values 4elow 3.3 an# no #ye@4in#in5 measure@ ments coul# 4e o4taine#. 9"e e!!ects o! #ye concentration an# c"itin/ c"itosan #ye solution ratios on #ye@4in#in5 ca$acity an# water u$ta/e o! c"itin an# c"itosan are #iscusse# in #etail elsew"ere B114C. &ar/e# #i!!erences 4etween water u$ta/e o! c"itin an# c"itosan e;ist wit" c"itosan ta/in5 more water t"an c"itin. 9"e #i!!erence may 4e #ue to #i!!erences in crystallinity o! t"e $ro#ucts or #ue to #i!!erences in t"e amount o! salt@!ormin5 5rou$s B113C. +i!!erences in t"e amount o! covalently 4oun# $rotein resi#ue mi5"t also a!!ect water u$ta/e. +ye concentrations "a# no mar/e# e!!ect on t"e water u$ta/e 4ut correlate# si5ni7cantly wit" t"e #ye@4in#in5 ca$acity o! c"itin an# c"itosan B116C. 9"e e!!ect o! $? on t"e #ye@ 4in#in5 ca$acity o! c"itin an# c"itosan was also stu#ie#. #ecline in t"e #ye@4in#in5 ca$acity a4ove $? 7.0 was o4serve#. Lit"in t"e $?

6.7. Pa$er 7nis"in5


,"itosan "as 4een re$orte# to im$art wet stren5t" to $a$er B117C. ?y#ro;ymet"yl c"itin an# ot"er water@solu4le #erivatives are use!ul en# a##itives in $a$er ma/in5. 9"is $olymeralt"ou5" $otentially availa4le in lar5e 8uan@ tities- never 4ecame a commercially si5ni7cant $ro#uct. 9"e entre$reneur in $a$er ma/in5 can utili6e t"is $olymer !or 4etter 7nis" $a$er $ro$erties.

6.D. =oli#@state 4atteries


,"itosan is insolu4le in water. 9"is $oses a $ro4lem in t"e !a4rication o! soli#@state $roton@ con#uctin5 4atteries 4ecause t"ere will not 4e any water $resent in t"e c"itosan w"ic" can act as a source o! "y#ro5en ions. 0n ot"er wor#st"e $roton@con#uctin5 $olymer nee#e# !or soli#@ state 4attery a$$lication cannot 4e o4taine# !rom c"itosan alone. ,"itosan is a 4io$olymer w"ic" can $rovi#e ionic con#uctivity w"en #issolve# in acetic aci#. 9"e con#uctivity is #ue to t"e $resence o! $rotons !rom t"e acetic aci# solution. 9"e trans$ort o! t"ese $rotons is t"ou5"t to occur t"rou5" many microvoi#s in t"e $olymer since t"e #ielectric constants !rom $ie6oelectric stu#ies are small. 9"e c"oice o! a more suita4le electro#e material may $ro#uce a 4etter 4attery system B11DC.

6.E. +ru5@#elivery systems


,ontrolle#@release tec"nolo5y emer5e# #ur@ in5 t"e 1ED0s as a commercially soun# met"@ o#olo5y. 9"e ac"ievement o! $re#icta4le an# re$ro#uci4le release o! an a5ent into a s$eci7c environment over an e;ten#e# $erio# o! time "as muc" si5ni7cant merit. 0t creates a #esire# environment wit" o$timal res$onse- minimum si#e@e!!ects an# $rolon5e# e!7cacy. ,ontrolle#@

14

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

release #osa5e !orms en"ance t"e sa!ety- e!7ca@ cy an# relia4ility o! #ru5 t"era$y. 9"ey re5ulate t"e #ru5 release rate an# re#uce t"e !re8uency o! #ru5 a#ministration to encoura5e $atients to com$ly wit" #osin5 instructions. ,onventional #osa5e !orms o!ten lea# to wi#e swin5s in serum #ru5 concentrations. &ost o! t"e #ru5 content is release# soon a!ter a#ministrationcausin5 #ru5 levels in t"e 4o#y to rise ra$i#ly$ea/ an# t"en #ecline s"ar$ly. For #ru5s w"ose actions correlate wit" t"eir serum #ru5 con@ centration- t"e s"ar$ Kuctuations o!ten cause unacce$ta4le si#e@e!!ects at t"e $ea/s- !ollowe# 4y ina#e8uate t"era$y at t"e trou5"s (Fi5. 2) B11EC. new #imension is t"e incor$oration o! 4io#e5ra#a4ility into t"e system. num4er o! #e5ra#a4le $olymers are $otentially use!ul !or t"is $ur$ose- inclu#in5 synt"etic as well as natural su4stances B11E1G2C. 9"e release o! #ru5s- a4sor4e# or enca$sulate# 4y $olymersinvolves t"eir slow an# controlla4le #i!!usion !rom/t"rou5" $olymeric materials. Pro#uction o! slow release (=R) #ru5s 4y t"e $"arma@ ceutical in#ustry is now a matter o! routine. +ru5s covalently attac"e# to 4io#e5ra#a4le $olymers or #is$erse# in a $olymeric matri; o! suc" macromolecules may 4e release# 4y ero@ sion/#e5ra#ation o! t"e $olymer. 9"era$eutic molecules- com$le;e# 4y $olymers- may also 4e release# !rom 5els 4y #i!!usion. ,"itosan is non@to;ic an# easily 4ioa4sor4@ a4le B74C wit" 5el@!ormin5 a4ility at low $?. &oreover- c"itosan "as antaci# an# antiulcer activities w"ic" $revent or wea/en #ru5 irrita@

tion in t"e stomac" B1GG-1G4C. lso- c"itosan matri; !ormulations a$$ear to Koat an# 5ra#ual@ ly swell in an aci# me#ium. ll t"ese interestin5 $ro$erties o! c"itosan ma/e t"is natural $olymer an i#eal can#i#ate !or controlle# #ru5 release !ormulations. &any e;cellent reviews an# 4oo/s #eal wit" t"e $ro$erties- c"emistry- 4ioc"emis@ try an# a$$lications o! c"itin- c"itosan an# t"eir #erivatives B1-4-34-72-7G-73-1G3-1G6C.

6.E.1. ?y#ro5els 4ase# on c"itin an# c"itosan ?y#ro5els are "i5"ly swollen- "y#ro$"ilic $olymer networ/s t"at can a4sor4 lar5e amounts o! water an# #rastically increase in volume. 0t is well /nown t"at t"e $"ysicoc"emical $ro$erties o! t"e "y#ro5el #e$en# not only on t"e molecu@ lar structure- t"e 5el structure- an# t"e #e5ree o! crosslin/in5- 4ut also on t"e content an# state o! t"e water in t"e "y#ro5el. ?y#ro5els "ave 4een wi#ely use# in controlle#@release systems B1G7-1GDC. Recently- "y#ro5els w"ic" swell an# contract in res$onse to e;ternal $? B1GE141C "ave 4een e;$lore#. 9"e $?@sensitive "y#ro5els "ave $o@ tential use in site@s$eci7c #elivery o! #ru5s to s$eci7c re5ions o! t"e 5astrointestinal tract (M0) an# "ave 4een $re$are# !or low molecular wei5"t an# $rotein #ru5 #elivery B142C. 0t is /nown t"at t"e release o! #ru5s !rom "y#ro5els #e$en#s on t"eir structure or t"eir c"emical $ro$erties in res$onse to $? B14G-144C. 9"ese $olymers- in certain cases- are e;$ecte# to resi#e in t"e 4o#y !or a lon5er $erio# an# res$on# to local environmental stimuli to mo#u@ late #ru5 release B143C. =ometimes t"e $olymers use# are 4io#e5ra#a4le to o4tain a #esira4le #evice to control #ru5 release B146C. 9"us- to 4e a4le to #esi5n "y#ro5els !or a $articular a$$lica@ tion- it is im$ortant to /now t"e nature o! t"e systems in t"eir environmental con#itions. =ome recent a#vances in controlle#@release !ormula@ tions usin5 5els o! c"itin an# c"itosan are $resente# "ere. 6.E.1.1. ,"itosan/$olyet"er inter$enetratin5 $olymer networ/ (0P() "y#ro5el Sao et al. B147C re$orte# a $roce#ure !or t"e $re$aration o! semi@0P( "y#ro5el 4ase# on

Fi5. 2. ,ontrolle# #ru5 #elivery versus imme#iate release.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

13

5lutaral#e"y#e@crosslin/e# c"itosan wit" an in@ ter$enetratin5 $olyet"er $olymer networ/. 9"e $? sensitivity- swellin5 an# release /inetics an# structural c"an5es o! t"e 5el in #i!!erent $? solutions were stu#ie# B1GE-140-14D-14EC. 9"e $"ysicoc"emical $ro$erties o! t"e "y#ro5el #e@ $en# not only on t"e molecular structure- t"e 5el structure an# t"e #e5ree o! crosslin/in5- 4ut also on t"e content an# state o! t"e water in t"e "y#ro5el. =ince t"e inclusion o! water si5ni7@ cantly a!!ects t"e $er!ormance o! "y#ro5els- a stu#y o! t"e $"ysical state o! water in t"e "y#ro5els is o! 5reat im$ortance to un#erstan# t"e nature o! interactions 4etween a4sor4e# water an# $olymers. Sao et al. B14EC stu#ie# t"e #ynamic water a4sor$tion c"aracteristics- state o! water- correlation 4etween state o! water an# swellin5 /inetics o! c"itosan$olyet"er "y#ro@ 5els 4y a$$lyin5 tec"ni8ues suc" as +=, an# some novel tec"ni8ues suc" as $ositron anni"i@ lation li!e@time s$ectrosco$y. Sao et al. B140C o4serve# ra$i# "y#rolysis o! t"e 5el wit" #ecrease in t"e ionic stren5t"- i.e. a "i5"er #e5ree o! swellin5 in lower ionic stren5t" solution B74C. 9"e "y#rolysis o! t"e 5el can 4e controlle# 4y t"e amount o! crosslin/er a$$lie#. 9"e more crosslin/er a##e#- t"e "i5"er t"e crosslin/ #ensity o! semi@0P(s- w"ic" re@ sults in a lower #e5ree o! swellin5 an# slower "y#rolysis B140C. ,"lor"e;i#ini acetas an# ,imeti#ine were use# as mo#el #ru5s !or #ru5 release stu#ies. !ast swellin5 o! 5els results in "i5"er #ru5 release at $? , 6 in com$arison to t"at at $? . 6 B1GE-147C.

"y#ro5els as woun#@coverin5 materials an# also stu#ie# t"e #ru5 release 4e"aviour usin5 silver sul!a#ia6ine as a mo#el #ru5 B132C.

6.E.1.G. ?y#ro5els o! $oly(et"ylene 5lycol)@co@ $oly(lactone) #iacrylate macromers an# b @ c"itin Aee an# *im B13GC re$orte# a $roce#ure !or $re$arin5 $oly(esteret"erester) tri4loc/ co@ $olymers. 9"e synt"esis o! t"e tri4loc/ co$oly@ mers was carrie# out 4y 4ul/ $olymeri6ation usin5 low to;ic stannous octoate as catalyst or wit"out catalyst (Fi5. G). 0nvesti5ations o! t"e t"ermal an# mec"anical $ro$erties were carrie# out. )itamin - vitamin < an# ri4oKavin were use# as mo#el #ru5s B134-133C. ?owever- t"ere were no re$orts on swellin5 /inetics an# solu@ 4ility $arameters o! t"e 5els. 6.E.1.4. ?y#ro5els o! $oly(et"ylene 5lycol) macromer/ b @c"itosan 0n t"eir stu#ies on c"itosan !or 4iome#ical a$$lications- Aee et al. B136C re$orte# a $ro@ ce#ure !or $re$arin5 semi@0P( $olymer networ/ "y#ro5els com$ose# o! b @c"itosan an# $oly@ (et"ylene 5lycol) #iacrylate macromer. 9"e "y#ro5els were $re$are# 4y #issolvin5 a mi;@ ture o! P<M& an# b @c"itosan in a8ueous acetic

6.E.1.2. =emi@0P( "y#ro5el $olymer networ/s o! b @c"itin an# $oly(et"ylene 5lycol) macromer =emi@0P( $olymer networ/ "y#ro5els com@ $ose# o! b @c"itin an# $oly(et"ylene 5lycol) macromer were synt"esi6e# !or 4iome#ical a$@ $lications B130-131C. 9"e t"ermal an# mec"ani@ cal $ro$erties o! t"ese "y#ro5els "ave also 4een stu#ie#. 9"e tensile stren5t"s o! semi@0P(s in t"e swollen state were !oun# to 4e 4etween 1.G3 an# 2.41 &Pa- t"e "i5"est re$orte# values to #ate !or crosslin/e# "y#ro5els. 9"ey use# t"ese

Fi5. G. =ynt"etic sc"eme o! P<MA& or P<M,&/ 0P(s.

b @c"itin semi@

16

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

aci#. 9"e resultin5 mi;ture was t"en cast to 7lms- !ollowe# 4y su4se8uent crosslin/in5 wit" 2-2@#imet"o;y@2@$"enylaceto$"enone as non@ to;ic $"oto initiator 4y .) irra#iation. 9"ey stu#ie# t"e crystallinity an# t"ermal an# me@ c"anical $ro$erties o! t"e 5els.

6.E.1.3. ?y#ro5els o! c"itosan/5elatin "y4ri# $olymer networ/ Sao et al. B137C re$orte# a novel "y#ro5el 4ase# on crosslin/e# c"itosan/5elatin wit" a 5lutaral#e"y#e "y4ri# $olymer networ/. 9"ey o4serve# #rastic swellin5 o! t"e 5els at aci#ic $? in com$arison to 4asic solutions. Aevamisole- cimeti#ine an# c"loram$"enicol were use# as mo#el #ru5s. $?@#e$en#ent release o! cimeti#ine- levamisole an# c"loram@ $"enicol !rom t"e 5el was re$orte#. 6.E.1.6. ,"itosanamine o;i#e 5el +utta et al. BG0G2C $re$are# an "omo5e@ neous c"itosanamine o;i#e 5el an# stu#ie# its swellin5 4e"avior an# release c"aracteristics in a 4u!!er solution ($? 7.4) at room tem$erature. ?omo5enous erosion o! t"e matri; an# a near 6ero@or#er release o! am$icillin tri"y#rate were o4serve#. 9"ey re$orte# t"e t"ermal $ro$erties o! t"e c"itosanamine o;i#e 5el in a !urt"er stu#y BG1C. 6.E.2. ,"itin an# c"itosan ta4lets &any #irect@com$ression #iluents "ave 4een re$orte# in t"e literature- 4ut every #iluent "as some #isa#vanta5es B13DC. &icrocrystalline cel@ lulose (&,,) "as 4een wi#ely use# as a ta4let #iluent in 1a$an. ,"itin an# c"itosan- 4ecause o! t"eir versatility- "ave 4een re$orte# to 4e use!ul #iluents in $"armaceutical $re$arations B13E-160C. 6.E.2.1. +irectly com$resse# ta4lets containin5 c"itin or c"itosan in a##ition to lactose or $otato starc" =awayana5i et al. B161C re$orte# t"e Kui#ity an# com$ressi4ility o! com4ine# $ow#ers o! lactose wit" c"itin (lactose/c"itin)- wit"

c"itosan (lactose/c"itosan) an# $otato starc" wit" c"itin ($otato starc"/c"itin)- an# wit" c"itosan ($otato starc"/c"itosan). 9"e #isinte@ 5ration $ro$erties o! ta4lets ma#e !rom t"ese $ow#ers- in com$arison wit" t"ose o! com4ine# $ow#ers o! lactose wit" &,, (lactose/&,,) an# $otato starc" wit" &,, ($otato starc"/ &,,) in or#er to #evelo$ new #irect@com$res@ sion #iluents- are also re$orte# B161C. 9"e Kui#ity o! com4ine# $ow#ers wit" c"itin an# c"itosan was 5reater t"an t"at o! t"e $ow#er wit" crystalline cellulose. 9"e re$orte# "ar#ness o! t"e ta4lets !ollows t"e or#er> c"itosan ta4lets . &,, . c"itin. 0n #isinte5ration stu#iesta4lets containin5 less t"an 70F c"itin or c"itosan $asse# t"e test. &oreover- t"e e'ection !orce o! t"e ta4lets o! lactose/c"itin an# lac@ tose/c"itosan was si5ni7cantly less t"an t"at o! lactose/crystalline cellulose ta4lets B161C. ?ow@ ever- no re$orts are availa4le on controlle# #ru5 release !ormulations usin5 t"ese ta4lets.

6.E.2.2. ,"itosan ta4lets !or controlle# release> anioniccationic inter$olymer com$le; Recently- c"itosan "as 5aine# im$ortance as a #isinte5ration a5ent #ue to its stron5 a4ility to a4sor4 water. 0t "as 4een o4serve# t"at c"itosan containe# in ta4lets at levels 4elow 70F acts as a #isinte5ration a5ent B161-162C. (eau et al. B16GC investi5ate# t"e sustaine#@release c"arac@ teristics o! et"ylcellulose ta4lets containin5 t"eo$"ylline as t"e mo#el #ru5. =everal e8ua@ tions were teste# to c"aracteri6e release mec"a@ nisms wit" res$ect to t"e release #ata. 9"e investi5ations reveal t"at- at "i5" #ru5 loa#in5#ru5 was release# 4y a #i!!usion mec"anism wit" a rate constant t"at increase# wit" an increase in a8ueous solu4ility. t low #ru5 loa#in5- $olymer rela;ation also 4ecomes a com$onent o! t"e release mec"anism. ?oweverits contri4ution to #ru5 release was less $ro@ nounce# as #ru5 solu4ility #ecrease#- 4ecomin5 ne5li5i4le in t"e case o! t"eo$"ylline. Recently- &i et al. B164C "ave re$orte# al5inate as an anionic $olyelectrolyte to control t"e swellin5 an# erosion rates o! c"itosan ta4lets

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

17

in aci#ic me#ia. 0nvesti5ations o! t"e #ru5 release mec"anism o! various ta4lets "ave 4een carrie# out 4ase# on Pe$$asNs mo#el B163-166C an# nuclear ma5netic resonance ima5in5 micro@ sco$y was use# to e;amine t"e swellin5/#i!!u@ sion mec"anism o! various ta4lets B164C.

6.E.G. &icroca$sules/micros$"eres o! c"itosan Pmicroca$suleN is #e7ne# as a s$"erical $article wit" si6e varyin5 !rom 30 nm to 2 mmcontainin5 a core su4stance. &icros$"eres arein a strict sense- s$"erical em$ty $articles. ?owever- t"e terms microca$sules an# micro@ s$"eres are o!ten use# synonymously. 0n a##i@ tion- some relate# terms are use# as well. For e;am$le- Pmicro4ea#sN an# P4ea#sN are use# alternatively. =$"eres an# s$"erical $articles are also use# !or a lar5e si6e an# ri5i# mor$"olo5y. Recently- Sao et al. B167C "i5"li5"te# t"e $re$a@ ration an# $ro$erties o! microca$sules an# micros$"eres relate# to c"itosan. +ue to t"e attractive $ro$erties an# wi#er a$$lications o! c"itosan@4ase# microca$sules an# micros$"eresa survey o! t"e a$$lications in controlle# #ru5 release !ormulations is a$$ro$riate. &oreovermicroca$sule an# micros$"ere !orms "ave an e#5e over ot"er !orms in "an#lin5 an# a#minis@ tration. 6.E.G.1. ,rosslin/e# c"itosan micros$"eres coate# wit" $olysacc"ari#es or li$i# 9"e $re$aration o! crosslin/e# c"itosan micros$"eres coate# wit" $olysacc"ari#e or li$i# !or intelli5ent #ru5 #elivery systems "as 4een re$orte# (Fi5. 4) B167C. 9"e micros$"eres were $re$are# wit" an inverse emulsion o! 3@ Kuorouracil (3@F.) or its #erivative solution o! "y#roc"loric aci# o! c"itosan in toluene con@ tainin5 =P ( D0. ,"itosan was crosslin/e# t"rou5" =c"i!!Ns salt !ormation 4y a##in5 a 5lutaral#e"y#e solution in toluene. t t"e same time- t"e amino #erivatives o! 3@F. were immo4ili6e#- o4viously resultin5 in an increase in t"e amount o! #ru5 wit"in t"e micros$"eres. 9"e micros$"eres were coate# wit" anionic $olysacc"ari#es (e.5.- car4o;ymet"ylc"itin- etc.)

Fi5. 4. =c"ematic structure o! a c"itosan 5el micros$"ere coate# wit" anionic $olysacc"ari#e an# li$i#.

t"rou5" a $olyion com$le; !ormation reaction. 0n t"e case o! li$i#@coate# micros$"eres- t"e micros$"eres alon5 wit" #i$almitoyl $"os@ $"ali#yl c"oline (+PP,) were #is$erse# in c"loro!orm. !ter eva$oration o! t"e solventmicros$"eres were o4taine# coate# wit" a +PP, li$i# multilayer- w"ic" e;"i4ite# a transi@ tion tem$erature o! a li8ui# crystal $"ase at 41.48,. 9"e #iameter ran5e o! t"e micros$"eres was 230G00 nm wit" a narrow #istri4ution. 9"e sta4ility o! t"e #is$ersion was im$rove# 4y coatin5 (Fi5. 3) t"e micros$"ere wit" anionic $olysacc"ari#e or a li$i# multilayer. com$arative stu#y on t"e release o! 3@F. an# its #erivatives !rom a $olysacc"ari#e@coate# micros$"ere &= (,&) was carrie# out in $"ysiolo5ical saline at G7 8,. 9"e #ata in#icate# t"at t"e 3@F. release rate #ecrease# in t"e or#er> !ree 3@F. . car4o;ymet"yl ty$e 3@F. . ester ty$e 3@F.. 9"e results reveale# t"at t"e coatin5 layers on t"e micros$"eres were e!!ec@ tive 4arriers to 3@F. release. Ai$i# mutilayers wit" a "omo5eneous com@ $osition 5enerally s"ow a 5elli8ui# crystal transition. L"en t"e tem$erature is raise# to 428,- w"ic" is "i5"er t"an t"e $"ase transition o! 41.4 8,- t"e amount o! 3@F. release# in@ crease#- an# t"e amount o! #ru5 #elivere# #ecrease# at G7 8,- w"ic" is lower t"an t"e transition tem$erature. +ue to t"e im$rove# reco5nition !unction o! $olysacc"ari#e c"ains !or animal cell mem4ranes- #elivery systems !rom $olysacc"ari#e@coate# micros$"eres- &= (,&)- seem $romisin5.

1D

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

&oreover- t"e release rate can 4e controlle# via t"e com$osition o! t"e ?P( an# t"e #e5ree o! #eacetylation o! c"itosan.

6.E.G.G. ,"itosan micros$"eres !or controlle# release o! #iclo!enac so#ium Mo"el et al. B16EC re$orte# on c"itosan micro@ s$"eres containin5 #iclo!enac so#ium- w"ic" were $re$are# 4y a coacervation $"ase se$a@ ration met"o#. ,"itosan an# 5lutaral#e"y#e were use# as coatin5 material an# crosslin/in5 a5ent- res$ectively. 0n vivo stu#ies were $er@ !orme# on (ew Tealan# w"ite ra44its. &ore@ over- t"e micros$"eres were !oun# to 4e sta4le at 43 8, !or G0 #ays. =tu#entNs P tN test was $er!orme# !or t"e results o! in vitro #issolution #ata !or !res" an# a5e# sam$les (G0 #ays at 438,) an# no si5ni7cant #i!!erence was !oun# u$on stora5e.
Fi5. 3. Pre$aration o! &= (,&)- &= (,&A) an# &= (,&) $olysacc"ari#e. sc"ematic #ia5ram.

6.E.G.2. ,"itosan/5elatin networ/ $olymer micros$"eres 0n t"eir stu#ies on t"e $"armaceutical a$$li@ cations o! c"itin an# c"itosan- Sao an# co@ wor/ers B16DC re$orte# c"itosan/5elatin net@ wor/ $olymer micros$"eres !or controlle# re@ lease o! cimeti#ine. 9"e #ru5@loa#e# micro@ s$"eres were $re$are# 4y #issolvin5 c"itosan5elatin (1>1 4y wei5"t) an# cimeti#ine in 3F acetic aci#. certain amount o! 9ween@D0 an# li8ui# $ara!7n at a water@to@oil ratio o! 1>10 was a##e# to t"e c"itosan/5elatin mi;ture un#er a5itation at 630 r$m at G0 8,. suita4le amount o! 23F a8ueous 5lutaral#e"y#e was a##e# to t"e inverse emulsion an# t"e system maintaine# !or 2 ". Finally- t"e li8ui# $ara!7n was va$or@ i6e# un#er vacuum to o4tain micros$"eres. 9"e #ru5 release stu#ies were $er!orme# in "y#roc"loric aci# solution ($? 1.0) an# $otas@ sium #i"y#ro5en $"os$"ate ($? 7.D) 4u!!er at an ionic stren5t" o! 0.1 m/l. $?@#e$en#ent $ulse#@release 4e"avior o! t"e "y4ri# $olymer networ/ (?P() matri; was o4serve# B16DC.

6.E.G.4. ,"itosan$olyet"ylene o;i#e nano$articles as $rotein carriers ?y#ro$"ilic nano$articulate carriers "ave many $otential a$$lications !or t"e a#ministra@ tion o! t"era$eutic molecules. 9"e recently #evelo$e# "y#ro$"o4ic"y#ro$"ilic carriers re@ 8uire t"e use o! or5anic solvents !or t"eir $re$aration an# "ave a limite# $rotein@loa#in5 ca$acity B17017GC. 9o a##ress t"ese limita@ tions- ,alvo et al. B174C re$orte# a new a$@ $roac" !or t"e $re$aration o! nano$articles ma#e solely o! "y#ro$"ilic $olymer. 9"e $re$a@ ration tec"ni8ue- 4ase# on an ionic 5elation $rocess- is e;tremely mil# an# involves a mi;ture o! two a8ueous $"ases at room tem@ $erature (Fi5. 6). Jne $"ase contains t"e c"itosan (,=) an# a #i4loc/ co$olymer o! et"ylene o;i#e an# so#ium tri$oly$"os$"ate (9PP). 9"e si6e (2001000 nm) an# 6eta $o@ tential (4etween 1 20 an# 1 60 m)) o! t"e nano$articles can 4e mo#ulate# conventionally 4y varyin5 t"e ,=/P<J@PPJ ratio. Furt"er@ more- usin5 4ovine serum al4umin (:= ) as a mo#el $rotein- it was s"own t"at t"ese new nano$articles "ave 5reat $rotein loa#in5 ca$aci@ ty (entra$ment e!7ciency u$ to D0F o! t"e

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

1E

Fi5. 6. 9"e $re$aration o! ,= nano$articles. 5ram.

sc"ematic #ia@

GE0 000 c"itosan at $? 4 (less t"an 7F loss wit" re5ar# to t"e 130 5/l initial concentration). =imilarly- t"e enca$sulation o! various mole@ cules B"aemo5lo4in (?4)- 4ovine serum al4umin (:= ) an# #e;trans wit" various molecular wei5"tsC in calcium al5inate 4ea#s coate# wit" c"itosan "as 4een re$orte# B176-177C. 9"eir release "as 4een com$are# an# t"e inKuence o! t"e #imensions- t"e c"emical com$osition an# t"e molecular wei5"t o! t"e enca$sulate# ma@ terials "ave 4een analyse# B176C. 9"e ionic interactions 4etween al5inate an# c"itosan at #i!!erent $? are #e$icte# in Fi5. 7.

$rotein) an# can $rovi#e continuous release o! t"e entra$$e# $rotein !or u$ to 1 wee/.

6.E.G.3. ,"itosan/calcium al5inate 4ea#s 9"e enca$sulation $rocess o! c"itosan an# calcium al5inate as a$$lie# to enca$sulation o! "aemo5lo4in was re$orte# 4y ?u5uet et al. B173C. 0n t"e 7rst $rocess- a mi;ture o! "aemo@ 5lo4in an# so#ium al5inate is a##e# #ro$wise to a solution o! c"itosan an# t"e interior o! t"e ca$sules t"us !orme# in t"e $resence o! ,a,l is 2 "ar#ene#. 0n t"e secon# met"o#- t"e #ro$lets were #irectly $ulle# o!! in a c"itosan,a,l 2 mi;ture. :ot" $roce#ures lea# to 4ea#s con@ tainin5 a "i5" concentration o! "aemo5lo4in (more t"an E0F o! t"e initial concentration (130 5/l) is retaine# insi#e t"e 4ea#s) $rovi#e# t"e c"itosan concentration is su!7cient. 9"e molecular mass o! c"itosan (243 000 or GE0 000 +a) an# t"e $? (2- 4- or 3.4) "a# only a sli5"t e!!ect on t"e entra$ment o! "aemo@ 5lo4in- t"e 4est retention 4ein5 o4taine# wit" 4ea#s $re$are# at $? 3.4. 9"e release o! "aemo5lo4in #urin5 4ea# stora5e in water was !oun# to 4e #e$en#ent on t"e molecular wei5"t o! c"itosan. 9"e 4est retention #urin5 stora5e in water was o4taine# wit" 4ea#s $re$are# wit" a "i5" molecular wei5"t c"itosan solution at $? 2.0. ,onsi#erin5 t"e total loss in "aemo5lo4in #urin5 4ea# !ormation an# a!ter 1 mont" o! stora5e in water- t"e 4est results were o4taine# 4y $re$arin5 t"e 4ea#s in an D 5/l solution o!

6.E.G.6. &ulti$orous 4ea#s o! c"itosan =everal researc"ers B17D-17EC "ave stu#ie# sim$le coacervation o! c"itosan in t"e $ro#uct@ ion o! c"itosan 4ea#s. 0n 5eneral- c"itosan is #issolve# in a8ueous acetic aci# or !ormic aci#. .sin5 a com$resse# air no66le- t"is solution is 4lown into (aJ?- (aJ?met"anol- or et"yl@ ene#iamine solution to !orm coacervate #ro$s. 9"e #ro$s are t"en 7ltere# an# was"e# wit" "ot an# col# water successively. )aryin5 t"e e;clu@ sion rate o! t"e c"itosan solution or t"e no66le #iameter can control t"e #iameter o! t"e #ro$@ lets. 9"e $orosity an# stren5t" o! t"e 4ea#s corres$on# to t"e concentration o! t"e c"itosan aci# solution- t"e #e5ree o! (@#eacetylation o! c"itosan- an# t"e ty$e an# concentration o! coacervation a5ents use#. 9"e c"itosan 4ea#s #escri4e# a4ove "ave 4een a$$lie# in various 7el#s- vi6. en6ymatic immo4ili6ation- c"romato5ra$"ic su$$ort- a#@ sor4ent o! metal ions- or li$o$rotein- an# cell cultures. 0t was con7rme# t"at t"e $orous sur!aces o! t"e c"itosan 4ea#s !orm a 5oo# cell culture carrier. ?ayas"i an# 0/a#a B1D0C im@ mo4ili6e# $rotease onto $orous c"itosan 4ea#s wit" a s$acer an# !oun# t"at t"e immo4ili6e# $rotease "a# "i5"er $?- an# t"ermal stora5e sta4ility- an# e;"i4ite# "i5"er activity towar#s t"e small ester su4strate (@4en6yl@A@ar5inine et"yl ester. 0n a##ition- (is"imura et al. B17DC investi5ate# t"e $ossi4ilities o! usin5 c"itosan 4ea#s as a carrier !or t"e cancer c"emot"era@

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&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

"i5"er release rates at $? 12 t"an at $? 7.27.4. 9"e e!!ect o! t"e amount o! #ru5 loa#e#- t"e molecular wei5"t o! c"itosan an# t"e crosslin/in5 a5ent on t"e #ru5@#elivery $ro7les "ave 4een re$orte# B1D11DGC.

Fi5. 7. =c"ematic re$resentation o! t"e ionic interactions 4etween al5inate an# c"itosan> (a) $? 3.42 (4) $? 2.0.

$eutic a#riamycin. Recently- ="arma et al. B1D11DGC $re$are# c"itosan micro4ea#s !or oral sustaine# #elivery o! ne!e#i$ine- am$icillin an# various steroi#s 4y a##in5 t"ese #ru5s to c"itosan an# t"en enterin5 a sim$le coacerva@ tion $rocess. 9"ese coacervate 4ea#s can 4e "ar#ene# 4y crosslin/in5 wit" 5lutaral#e"y#e or e$o;yc"loro$ro$ane to $ro#uce microca$sules containin5 rotun#ine B167C. 9"e release $ro7les o! t"e #ru5s !rom all t"ese c"itosan #elivery systems were monitore# an# s"owe#- in 5eneral-

6.E.4. ,"itosan@4ase# trans#ermal #ru5 #elivery systems 9"ac"aro#i an# Rao B1D41D6C re$orte# $er@ meation@controlle# trans#ermal #ru5 #elivery systems (9+=) usin5 c"itosan. =tu#ies on $ro@ $ranolol "y#roc"lori#e ($ro$@?,l) #elivery sys@ tems usin5 various c"itosan mem4ranes wit" #i!!erent crosslin/ #ensities as #ru5 release controllin5 mem4ranes an# c"itosan 5el as t"e #ru5 reservoir "ave 4een $er!orme#. 9"e $"ysicoc"emical $ro$erties o! t"e mem4ranes "ave 4een c"aracteri6e# an# t"e $ermea4ility c"aracteristics o! t"ese mem4ranes to 4ot" li$o$"ilic an# "y#ro$"ilic #ru5s "ave 4een re$orte# B1D4-1D3C. 0n vitro evaluations o! t"e 9+= #evices w"ile su$$orte# on ra44it $inna s/in were carrie# out in mo#i7e# Fran6 #i!!u@ sion cells B1D6C. 9"e in vitro #ru5 release $ro7les s"owe# t"at all #evices release# $ro$@ ?,l in a relia4le- re$ro#uci4le manner. 9"e #ru5 release was si5ni7cantly re#uce# w"en crosslin/e# c"itosan mem4ranes were use# to re5ulate #ru5 release in t"e #evices. &oreovert"e #ru5 release rate was !oun# to #e$en# on t"e crosslin/ #ensity wit"in t"e mem4ranes. 0t was o4serve# t"at t"e #evice constructe# wit" a c"itosan mem4rane wit" a "i5" crosslin/ #en@ sity release# t"e minimum amount o! #ru5. 9"is is #ue to t"e #ecrease# $ermea4ility coe!7cient o! crosslin/e# mem4ranes resultin5 !rom t"e crosslin/ $oints. 6.10. :iotec"nolo5y 6.10.1. Pre$aration o! 4iotec"nolo5ical materials ,"itin "as two "y#ro;yl 5rou$s- w"ile c"itosan "as one amino 5rou$ an# two "y#ro;yl 5rou$s in t"e re$eatin5 "e;osami#e resi#ue. ,"emical mo#i7cation o! t"ese 5rou$s an# t"e

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

21

re5eneration reaction 5ives rise to various novel 4io!unctional macromolecular $ro#ucts "avin5 t"e ori5inal or5ani6ation or new ty$es o! or5ani@ 6ation.

6.10.2. ,ell@stimulatin5 materials 6.10.2.1. 0n $lants &ainly two /in#s o! e;tracellular c"itinase were !oun# in t"e normal cell sus$ension cul@ ture o! rice (Jry6a sativa A. var. 'a$onica cv. *os"i"i/ari). 0n t"e $resence o! a c"itin oli5o@ sacc"ari#e mi;ture (#e5ree o! $olymeri6ation 2D)- "owever- t"e e;tracellular c"itinase activi@ ty increase# a4out t"ree@!ol# over t"e control on secretin5 an a##itional e;tracellular new c"itin@ ase iso!orm. 9"ese t"ree c"itinase iso!orms are one 5rou$ o! $at"o5enesis@relate# (PR) $roteins in $lants B1D7C. =oya4eans were coate# wit" a t"in layer o! #e$olymeri6e# c"itin- car4o;ymet"yl (,&)@ c"itin an# "y#ro;yet"yl (?<)@c"itin- an# t"e see#s were culture# in t"e 7el#. 0t was o4serve# t"at t"e see# c"itinase increase# 1.32.0@!ol#t"e see# 5ermination rate increase# 4y 6F- t"e $o# num4er increase# 4y EF- t"e $lant #ry wei5"t increase# 4y DF- an# t"e cro$ yiel# also increase# 4y 1012F over t"e control B1DDC. +ressin5 wit" c"itin 7lms- s$on5es an# 74res en"ance# c"itinase activity in tree@4ar/ tissues aroun# woun#s u$ to !our@!ol# over t"e control. 9"e c"itin 7lms- w"ic" were im$lante# in or use# to #ress t"e tree@4ar/ tissues- were #i5este# wit"in 4 to 24 wee/s t"erea!ter an# were assimilate# into t"e woun#e# 4ar/ tissues. 9"e !ate o! (@acetyl@+@5lucosamine in $lant tissue is un/nown. P"enylalanine ammonia@lyase was stimulate# 4y treatment wit" c"itin- an# li5nin !ormation in t"e $lant increase#. s a resultwoun# "ealin5 was accelerate# B1D7C. 6.10.2.2. 0n animals <;tracellular lyso6yme activity was en"ance# in in vitro cultures o! several mammalian cells 4y treatment wit" c"itin an# its #erivatives. s a result- connective tissue !ormation was stimu@

late#- an# t"e sel!@#e!ence !unction a5ainst micro4ial in!ection was en"ance# at t"e cellular level. Jn t"e 4asis o! t"ese results- several c"itin an# c"itosan #ressin5 materials (#iscusse# in t"e !ore5oin5 sections) "ave 4een #evelo$e# commercially !or t"e "ealin5 treatment o! "uman an# animal woun#s.

6.10.G. nti4acterial a5ents 9"e 5rowt" o! <sc"eric"ia coli was in"i4ite# in t"e $resence o! more t"an 0.023F c"itosan. ,"itosan also in"i4ite# t"e 5rowt" o! Fusariumlternaria an# ?elmint"os$orium. 9"e cationic amino 5rou$s o! c"itosan $ro4a4ly 4in# to anionic 5rou$s o! t"ese microor5anisms- re@ sultin5 in 5rowt" in"i4ition B1DEC. 6.10.4. :loo# anti@coa5ulants ("e$arinoi#s) ,"itin an# c"itosan sul$"ates "ave 4loo# anticoa5ulant an# li$o$rotein li$ase (APA)@re@ leasin5 activities. ,"itin G-6@sul!ate s"owe# a4out two@!ol# anticoa5ulant activity an# 0.1@ !ol# APA@releasin5 activity over t"ose o! "e$arin2 t"e sul!ate #erivatives mi5"t 4e usa4le as "e$arinoi#s !or arti7cial 4loo# #ialysis B1D7C. 6.10.3. nti@t"ro4o5enic an# "aemostatic materials ,"itosan 74res were !oun# to 4e t"rom@ 4o5enic an# "aemostatic in an in vitro test- an# (@"e;anoyl an# (@octanoyl c"itosan 74res were anti@t"rom4o5enic. ,"itosan 74res can 4e use# as "aemostatic material2 (@"e;anoyl an# (@ octanoylc"itosan 74res are use# as anti@t"rom@ 4o5enic materials B1E0C. 6.11. ,"itosan as !at tra$$er
Jne o! t"e c"aracters in a recent movie- P9"e Full &ontyN- "a# a memora4le line> PP9"e less 0 eat- t"e !atter 0 5et.NN 0ts a $"enomenon t"at $la5ues many #ieters w"o eat less an# lose muscle instea# o! !at. s a result- t"eir meta4o@ lism slows #own an# it 4ecomes more an# more #i!7cult to control wei5"t. Fortunately- it is not too #i!7cult to lose t"e ri5"t stu!!- !at- w"ile

22

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127

im$rovin5 muscle tone- meta4olism an# "ealt". &any su$$lements can "el$ in t"e !at re#uction $rocess- inclu#in5 $yruvate an# c"itosan. Pyru@ vate- !oun# in re# a$$les- some ty$es o! c"eesean# re# wine- stimulates !at loss an# 4oosts e;ercise $er!ormance. ,"itosan attac"es itsel! to !at in t"e stomac" 4e!ore it is #i5este#- t"us tra$$in5 t"e !at an# $reventin5 its a4sor$tion 4y t"e #i5estive tract. Fat in turn 4in#s to t"e c"itosan 74re- !ormin5 a mass w"ic" t"e 4o#y cannot a4sor4- an# w"ic" is eliminate# 4y t"e 4o#y. ,"itosan 74re #i!!ers !rom ot"er 74res in t"at it $ossesses a $ositive ionic c"ar5e- w"ic" 5ives it t"e a4ility to 4on# c"emically wit" t"e ne5atively c"ar5e# li$i#s- !ats an# 4ile aci#s B1E11EGC. 7. ,onclusion ,"itin an# c"itosan "ave a wi#e ran5e o! a$$lications. 9"ey may 4e em$loye#- !or e;am@ $le- to solve numerous $ro4lems in environmen@ tal an# 4iome#ical en5ineerin5. ,"itin #eriva@ tives inclu#in5 $artially #eacetylate# c"itosan can 4e easily mol#e# to various !orms an# t"eir #erivatives are #i5este# in vivo 4y lyso6omal en6ymes. 9"us- it a$$ears t"at t"is material can 4e a most interestin5 can#i#ate !or use as a carrier o! a variety o! #ru5s !or controlle#@ release a$$lications. Aately- t"e trans#ermal a4sor$tion $romotin5 c"aracteristics o! c"itosan "ave 4een e;$loite#- es$ecially !or nasal an# oral #elivery o! $olar #ru5s to inclu#e $e$ti#es an# $roteins an# !or vaccine #elivery. 9"ese $ro$erties- to5et"er wit" t"e very sa!e to;icity $ro7le- ma/e c"itosan an e;citin5 an# $romis@ in5 e;ci$ient !or t"e $"armaceutical in#ustry !or $resent an# !uture a$$lications. 9"an/s to t"e 4ioactivities o! c"itosan itsel!- its !ormulations wit" #ru5s may "ave #ual t"era$eutic e!!ects. c/nowle#5ements 9"e aut"or t"an/s +r *.M. Ramac"an#ran (air- ,entral 0nstitute o! Fis"eries 9ec"nolo5y-

*oc"i- 0n#ia- !or $rovi#in5 t"e sam$le o! c"itosan. 9"e aut"or is 5rate!ul to t"e ,ouncil o! =cienti7c an# 0n#ustrial Researc" (,=0R)&inistry o! ?uman Resource +evelo$ment Mrou$s- Movt. o! 0n#ia- (ew +el"i- !or 7nancial assistance to carry out t"is researc". 9"e aut"or is in#e4te# to t"e re!erees !or a t"orou5" revision o! t"e manuscri$t. Re!erences
B1C R. . . &u66arelli (<#.)- (atural ,"elatin5 Polymers- Per@ 5amon Press- (ew Sor/- 1E7G- $. DG. B2C 1.P. Ti/a/is (<#.)- ,"itin- ,"itosan an# Relate# <n6ymesca#emic Press- Jrlan#o- 1ED4- $. Q)00. BGC L. . &ass- . &ass- :. 9i5"e- review o! 4io#e5ra#a4le $olymers> uses- current #evelo$ments in t"e synt"esis an# c"aracteri6ation o! 4io#e5ra#a4le $olyesters. :len#s o! 4io@ #e5ra#a4le $olymers an# recent a#vances in 4io#e5ra#ation stu#ies- Polym. 0nt. 47 (1EED) DE. B4C A. 0llum- ,"itosan an# its use as $"armaceutical e;ci$ientP"arm. Res. 13 (1EED) 1G26. B3C =. (is"imura- J. *o"5o- *. *urita- ?. *u6u"ara- ,"emos@ $eci7c mani$ulations o! a ri5i# $olysacc"ari#e synt"esis o! novel c"itosan #erivatives wit" e;cellent solu4ility in com@ mon or5anic solvents 4y re5ioselective c"emical mo#i7ca@ tions- &acromolecules 24 (1EE1) 4743. B6C . 9o!!ey- M. =amaranaya/e- ,.<. Fra6ier- L.M. Mlasser,"itin #erivatives. 0. *inetics o! t"e "eat in#uce# conversion o! c"itosan to c"itin- 1. $$l. Polym. =ci. 1G (1EE6) 73. B7C =.1. *im- =.=. *im- S.&. Aee- =ynt"esis an# c"aracteri6ation o! et"er ty$e c"itin #erivatives- &acromol. ,"em. P"ys. 1E3 (1EE4) 16D7. BDC M. ,rini- M. 9orri- &. Muerrini- &. &orcellet- &. Leltrow@ s/i- :. &artel- (&R c"aracteri6ation o! (@4en6yl sul!onate# #erivatives o! c"itosan- ,ar4o"y#r. Polym. GG (1EE7) 143. BEC &. =a/amota- =olvents o! cellulose an# c"itin an# t"eir a$$lication- in> Procee#in5s o! t"e Gr# sian 9e;tile ,on!er@ ence- 1EE3- $. GE. B10C 1.T. *naul- *. .&. ,re4er- ,oa5ulation rate stu#ies o! s$inna4le c"itosan solutions- 1. $$l. Polym. =ci. 66 (1EE7) 117. B11C &. =a/amoto- ?. 9sen5- *. Furu"ata- Re5ioselective c"lori@ nation o! c"itin wit" (@c"lorosuccinimi#etri$"enyl$"os@ $"ine un#er "omo5eneous con#itions in lit"ium c"lori#e (-(@#imet"ylacetami#e- ,ar4o"y#r. Res. 263 (1EE4) 271. B12C ?. 9sen5- *. Furu"ata- &. =a/amoto- :romination o! re5enerate# c"itin wit" (@4romosuccinimi#e an# tri@ $"enyl$"os$"ine un#er "omo5eneous con#itions in lit"ium 4romi#e(-(@#imet"ylacetami#e- ,ar4o"y#r. Res. 270 (1EE3) 14E. B1GC ?. 9sen5- R. Aee- *. Furu"ata- &. =a/amoto- :romination o! c"itin wit" tri4romoimi#a6ole an# tri$"enyl$"os$"ine in lit"ium 4romi#e#imet"ylacetami#e- =en@0 Ma//ais"i 31 (1EE3) 340.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 B14C *. *urita- ?. Sos"ino- *. So/ota- &. n#o- =. 0noue- =. 0s"ii- =. (is"imura- Pre$aration o! tosyl c"itins as $recursors !or !acile c"emical mo#i7cation o! c"itin- &acromolecules 23 (1EE2) G7D6. B13C *. *urita- =. 0noue- =. (is"imura- Pre$aration o! solu4le c"itin #erivatives as reactive $recursors !or controlle# mo#i@ 7cations> tosyl@ an# io#o c"itins- 1. Polym. =ci.- Part > Polym. ,"em. 2E (1EE1) EG7. B16C +.). Auyen- ). Ross4ac"- &i;e# esters o! c"itin- 1. $$l. Polym. =ci. 33 (1EE3) 67E. B17C &.9. Uuras"i- ?.=. :lair- =.1. llen- =tu#ies on mo#i7e# c"itosan mem4ranes- 0- 1. $$l. Polym. =ci. 46 (1EE2) 233. B1DC &.9. Uuras"i- ?.=. :lair- =.1. llen- =tu#ies on mo#i7e# c"itosan mem4ranes. 00. +ialysis o! low molecular wei5"t meta4olites- 1. $$l. Polym. =ci. 46 (1EE2) 26G. B1EC (. *u4ota- Permeation $ro$erties o! c"itosan transition metal com$le; mem4ranes- 1. $$l. Polym. =ci. 64 (1EE7) D1E. B20C =. ?irano- !acile met"o# !or t"e $re$aration o! novel mem4ranes !rom (@acyl an# (@aryli#ene c"itosan 5els5ric. :iol. ,"em. 14 (1E7D) 1EGD. B21C M.L. .r4anc6y/- :. Ai$$@=ymonowic6- 9"e inKuence o! $rocessin5 terms o! c"itosan mem4ranes ma#e o! #i!!erently #eacetylate# c"itosan on t"e crystalline structure o! mem@ 4ranes- 1. $$l. Polym. =ci. 31 (1EE4) 21E1. B22C *. *urita- *. 9omita- =. 0s"ii- =. (is"imura- *. ="imo#ab @,"itin as a convenient startin5 material !or acetolysis !or e!7cient $re$aration o! (@acetylc"itooli5osacc"ari#es- 1. Polym. =ci.- Part > Polym. ,"em. G1 (1EEG) 2GEG. B2GC *.R. ?olme- A.+. ?all- ,"itosan #erivatives 4earin5 , 10 @ al/yl 5lycosi#e 4ranc"es> a tem$erature in#uce# 5ellin5 Polysacc"ari#e- &acromolecules 24 (1EE1) GD2D. B24C P. &a#"avan (<#.)- ,"itin- ,"itosan an# t"eir (ovel $$li@ cations- =cience Aecture =eries- ,0F9- *oc"i- 1EE2- $. 1. B23C R. . . &u66arelli- in> ?.F. &ar/- (.&. :i/ales- ,.M. Jver4er5er- M. &en5es (<#s.)- 2n# <#ition- <ncyclo$e#ia o! Polymer =cience an# <n5ineerin5- )ol. G- Liley- (ew Sor/1ED4- $. 4G3. B26C P. &a#"avan- *.M.R. (air- .tili6ation o! $rawn waste. 0solation o! c"itin an# its conversion to c"itosan- Fis"ery 9ec". 11 (1E74) 30. B27C P. &a#"avan- *.M.R. (air- 9.*. 9"an/a$$an- P.). Pra4"u- *. Mo$a/umar- Pro#uction o! c"itin an# c"itosan- Pro'ect Re$ort- ,0F9- *oc"i (Price# :ulletin)- 1ED6. B2DC *. Mo$a/umar- P.M.). (air- &.*. &u/un#am- 1. =te$"en.M. Ra#"a/ris"nan- *.M.R. (air- P.+. nt"ony et al.:ioc"emical com$osition o! 0n#ian !oo# 7s"- ,0F9 :ulletin1EE7. B2EC A. ?enc" Aarry- :iomaterials> a !orecast !or t"e !uture:iomaterials 1E (1EED) 141E. BG0C P.*. +utta- P. )iswanat"an- A. &imrot- &.(.). Ravi *umar.se o! c"itosan amine o;i#e 5el as #ru5 carrier- 1. Polym. &ater. 14 (1EE7) G31. BG1C P.*. +utta- &.(.). Ravi *umar- ,"itosanamine o;i#e> t"ermal 4e"avior o! new 5ellin5 system- 0n#ian 1. ,"em. 9ec"nol. 6 (1EEE) 33. BG2C &.(.). Ravi *umar- P. =in5"- P.*. +utta- <!!ect o! swellin5 on c"itosanamine o;i#e 5el in e;ten#e# #ru5 #elivery0n#ian +ru5s G6 (1EEE) GEG. BGGC (. (is"i- 1. (o5uc"i- =. 9o/ura- ?. ="iota- =tu#ies on c"itin. 0. cetylation o! c"itin- Polym. 1. 11 (1E7E) 27.

2G

BG4C *. *ai!u- (. (is"i- 9. *omai- Pre$aration o! "e;anoyl#ecanoyl an# #o#ecanoyl c"itin- 1. Polym. =ci.- Part > Polym. ,"em. 1E (1ED1) 2G61. BG3C . :a;ter- &. +illon- *.+. . 9aylor- M. .F. Ro4erts0m$rove# met"o# !or 0R #etermination o! t"e #e5ree o! (@acetylation o! c"itosan- 0nt. 1. :iol. &acromol. 14 (1EE2) 166. BG6C M. . &a5"ami- M. .F. Ro4erts- =tu#ies on t"e a#sor$tion o! anionic #yes on c"itosan- &a/romol. ,"em. 1DE (1EDD) 22GE. BG7C . +omar#- ,ircular #ic"roism stu#y on (@ acetyl5lucosamine oli5omers- 0nt. 1. :iol. &acromol. D (1ED6) 24G. BGDC . +omar#- $? an# c.#. measurements on a !ully 00 #eacetylate# c"itosan> a$$lication to ,u $olymer interac@ tions- 0nt. 1. :iol. &acromol. E (1ED7) ED. BGEC S.,. Lei- =.&. ?u#son- :in#in5 o! so#ium #o#ecyl sul!ate to a $olyelectrolyte 4ase# c"itosan- &acromolecules 26 (1EEG) 4131. B40C ?. =as"iwa- ?. =aimoto- S. ="i5emasa- R. J5awa- =. 9o/ura- +istri4ution o! t"e acetami#o 5rou$ in $artially #eacetylate# c"itins- ,ar4o"y#r. Polym. 16 (1EE1) 2E1. B41C ?. =as"iwa- ?. =aimoto- S. ="i5emasa- =. 9o/ura- (@ cetyl 5rou$ #istri4ution in $artially #eacetylate# c"itins $re$are# un#er "omo5eneous con#itions- ,ar4o"y#r. Res. 242 (1EEG) 167. B42C A. Raymon#- F.M. &orin- R.?. &arc"essault- +e5ree o! #eacetylation o! c"itosan usin5 con#uctometric titration an# soli#@state (&R- ,ar4o"y#r. Res. 24G (1EEG) GG1. B4GC F. (iola- (. :asora- <. ,"ornet- P.F. )i#al- ra$i# met"o# !or t"e #etermination o! t"e #e5ree o! (@acetylation o! c"itinc"itosan sam$le 4y aci# "y#rolysis an# ?PA,- 1. 9"erm. nal. 2D (1EDG) 1DE. B44C =.?. Pan54urn- P.). 9rescony- 1. ?eller- in> 1.P. Ti/a/is (<#.),"itin- ,"itosan an# Relate# <n6ymes- ?arcourt :race 1anovic"- (ew Sor/- 1ED4- $. G. B43C 9.+. Rat"/e- =.&. ?u#son- +etermination o! t"e #e5ree o! (@#eacetylation in c"itin an# c"itosan as well as t"eir monomer su5ar ratios 4y near in!rare# s$ectrosco$y- 1. Polym. =ci.- Polym. ,"em. <#. G1 (1EEG) 74E. B46C .,.&. Lu- +etermination o! molecular wei5"t #istri4ution o! c"itosan 4y "i5"@$er!ormance li8ui# c"romato5ra$"y&et"o#s <n6ymol. 161 (1EDD) 447. B47C R. . . &u66arelli- ,. Aou5"- &. <manuelli- 9"e molecular wei5"t o! c"itosan stu#ie# 4y laser li5"t scatterin5- ,ar@ 4o"y#r. Res. 164 (1ED7) 4GG. B4DC ).F. Aee- =olution an# s"ear $ro$erties o! c"itin an# c"itosan- P".+. +issertation- .niversity o! Las"in5ton.niversity &icro7lms- nn r4or- &0- .= - &icro7lm 74@2E- 1E74- $. 446. B4EC P.P. von Leimarn- 1. 9e;t. 0nst. 17 (1E26) 9642. B30C M. *uni/e,"itin un# c"itinsei#e*unsi#ei#e (,"emie!asern) D (1E26) 1D2. B31C M.A. ,lar/- .F. =mit"- Q@ray #i!!raction stu#ies o! c"itinc"itosan an# #erivatives- 1. P"ys. ,"em. 40 (1EG6) D6G. B32C =.&. ?o#son- 1. . ,uculo- 9"e solu4ility o! unmo#i7e# cellulose> a criti8ue o! t"e literature- 1.&.=.@Rev. &acromol. ,"em. ,1D (1ED0) 1. B3GC 9.R. +awsey- ,.A. &c ,ormic/- 9"e lit"ium/#imet"yl@ acetami#e solvent !or cellulose> a literature review- 1.&.=.@ Rev. &acromol. ,"em. P"ys. ,G0 (1EE0) 403.

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&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 B76C R. . . (u66arelli- ,. &u66arelli- &. 9er4o'evic"- ,"itin c"emistry u$5ra#in5 a renewa4le resource- ,ar4o"y#r. <ur. 1E (1EE7). B77C &.(.). Ravi *umar- P.*. +utta- =. (a/amura- &et"o#s o! metal ca$ture !rom wastewater- in> R.*. 9rive#y (<#.)#vances in Lastewater 9reatment 9ec"nolo5ies- Mlo4al =cience- 0n#ia- 1EED- $. 22. B7DC ?.F. &ar/- (.&. :i/ales- ,.M. Jver4er5er- M. &en5es (<#s.)- <ncyclo$e#ia o! Polymer =cience an# <n5ineerin5)ol. 1- Liley- (ew Sor/- 1ED3- $. 20. B7EC L.M. &alettas- ?.1. Uuin5ley- <.+. #ic/es- in> R. . . &u66arelli (<#.)- ,"itin in (ature an# 9ec"nolo5y- Plenum Press- (ew Sor/- 1ED6- $. 4G3. BD0C 0.). Sannas- ?.F. :ur/e- +.P. Jr5ill- <.&. =/ra4ut- Loun# tissue can utilise a $olymeric tem$late to synt"esise a !unctional e;tension o! s/in- =cience 213 (1ED2) 174. BD1C S. Ae- =.,. nan#- .R. ?orroc/s- +evelo$ment o! anti@ 4acterial $olysacc"ari#e 74res an# t"eir $er!ormance- in> <uro$ean ,on!erence on #vances in Loun# &ana5ementmster#am- (et"erlan#s- Jcto4er- 1EE6. BD2C R. Jlsen- +. =c"wart6miller- L. Le$$ner- R. Linan#y- in> M. =/'a/@:rac/- 9. nt"onsen- P. . =an#!or# (<#s.)- ,"itin an# ,"itosan> =ources- ,"emistry- :ioc"emistry- P"ysical Pro$@ erties an# $$lications- <lsevier $$lie# =cience- (ew Sor/- 1EDE. BDGC +. . =an#!or#- . =tinnes (<#s.)- :iome#ical $$lications o! ?i5" Purity ,"itosan R P"ysical- ,"emical an# :ioactive Pro$erties- ,= =ym$osium =eries- )ol. 467- 1EE1- $. 4G0. BD4C S. Ae- =.,. nan#- .R. ?orroc/s- Recent #evelo$ments in 74res an# materials !or woun# mana5ement- 0n#ian 1. Fi4re 9e;tile Res. 22 (1EE7) GG7. BD3C :. =$ar/es- +.M. &urray- .= Patent 4372E06 (1ED6). BD6C *. (ara- S. Sama5uc"i- ?. 9ane- .= Patent 4631723 (1ED7). BD7C *. *i!une- S. Sama5uc"i- =. *is"imoto- 9rans. =oc. :ioma@ ter. Q0 (1EDD) 216. BDDC *.S. *im- +.=. &in- 9rans. =oc. :iomater. Q0 (1EDD) 63D. BDEC M. :ia5ini- . :ertani- R. &u66arelli et al.- ,ar4o"y#r. Res. 12 (1EE1) 2D1. BE0C :. =a5ar- P. ?amlyn- +. Lales- <ur. Patent 0460774 2 (1EE1). BE1C :. =a5ar- P. ?amlyn- +. Lales- .* Patent 21DDD1G3 (1ED7). BE2C :. =a5ar- P. ?amlyn- +. Lales- .* Patent 214DE3E (1ED3). BEGC :. =a5ar- P. ?amlyn- +. Lales- .* Patent 2163D63 (1ED6). BE4C R. . . &u66arelli- .= Patent 3G7D472 (1EE3). BE3C :. =6oslan#- M.,. <ast- 9"e #ry s$innin5 o! #i4utyrylc"itin 74res- 1. $$l. Polym. =ci. 3D (1EE3) 243E. BE6C +. *norr- Recovery an# utili6ation o! c"itin an# c"itosan in !oo# $rocessin5 waste mana5ement- Foo# 9ec"nol. 1an. (1EE1) 114. BE7C =. (icol- Ai!e a!ter #eat" !or em$ty s"ells- (ew =cientist Fe4. (1EE1) 46. BEDC P.R. ustin- ,.1. :rine- 1.<. ,astle- 1.P. Ti/a/is- =cience 212 (1EDE) 74E. BEEC *. . =$reen- 1.P. Ti/a/is- P.R. ustin- in> 1.P. Ti/a/is (<#.),"itin- ,"itosan an# Relate# <n6ymes- ca#emic PressJrlan#o- 1ED4- $. 37. B100C 1.P. Ti/a/is- L.L. =aylor- P.R. ustin (<#s.)- ,"itin an# ,"itosan- 9"e 1a$anese =ociety o! ,"itin an# ,"itosan9ottori- 1ED2- $. 2GG. B101C &.A. &ar/ey- &.A. :owman- &.).L. :er5amini (<#s.),"itin an# ,"itosan- <lsevier $$lie# =cience- Aon#on1EDE- $. 71G.

B34C 9.+. Rat"/e- =.&. ?o#son- Review o! c"itin an# c"itosan as 74re an# 7lm !ormers- 1.&.=.@Rev. &acromol. ,"em. ,G4 (1EE4) G73. B33C P.R. ustin- .= Patent GDE27G1 (1E73). B36C ,.1. :rine- P.R. ustin- Renaturate# c"itin 74rils- 7lms an# 7laments- in> 9.+. ,"urc" (<#.)- &arine ,"emistry in ,oastal <nvironment- ,= =ym$osium =eries-)ol. 1D- ,=Las"in5ton- +,- 1E73- $. 303. B37C *. *i!une- *. 0nome- =. &ori- .= Patent 4EG2404 (1EE0). B3DC &. (a/a'ima- *. tsumi- *. *i!une- +evelo$ment o! a4sor4a4le sutures !rom c"itin- in> 1.P. Ti/a/is (<#.)- ,"itin,"itosan an# Relate# <n6ymes- ?arcourt :race 1anovic"(ew Sor/- 1ED4- $. 407. B3EC Fu'i =$innin5 ,o.- At#.- 1a$anese Patent 3E11641D (1ED4). B60C =. 9o/ura- (. (is"i- 1. (o5uc"i- =tu#ies on c"itin 000> Pre$aration o! c"itin 74res- Polym. 1. (9o/yo) 11 (1E7E) 7D1. B61C .niti/a ,o.- At#.- 1a$anese Patent 371GE101 (1ED2)2 ,"em. 4str.- ED(D)> 3344E5. B62C .niti/a ,o.- At#.- 1a$anese Patent 371GE101 (1ED2)2 ,"em. 4str.- ED(14)> 11G74$. B6GC .niti/a ,o.- At#.- 1a$anese Patent 3D21431G (1EDG)2 ,"em. 4str.- 100(22)> 1D01GDr. B64C .niti/a ,o.- At#.- 1a$anese Patent 3D214312 (1EDG)2 ,"em. 4str.- 100(22)> 1D01GEs. B63C F. . Rut"er!or# 000- P.R. ustin- &arine c"itin $ro$erties an# solvents- in> R. . . &u66arelli- <.R. Pariser (<#s.)- Procee#@ in5s o! t"e First 0nternational ,on!erence on ,"itin an# ,"itosan- :oston- & - &09 =ea Mrant Pro5ram- ,am4ri#5e1E77- $. 1D2. B66C P.R. ustin- ,"itin solvents an# solu4ility $arameters- in> 1.P. Ti/a/is (<#.)- ,"itin- ,"itosan an# Relate# <n6ymes- ?ar@ court :race 1anovic"- (ew Sor/- 1ED4- $. 227. B67C 9.<. =u/"anova- .). =i#orovic"- M.0. Moryainov- M.&. &i/"ailov- &. (itter$a/"ova- )yso/omol. =oe#in. =er. : G1 (1EDE) GD12 ,"em. 4str.- 111(20)> 173ED3n. B6DC A. . (u#N5a- Su.M. :a/la5ina- M. . Petro$avlovs/ii- M.0. Moryainov- M.&. &i/"ailov- A. . *o$ytc"en/o- )yso/omol. =oe#in. =er. : GG (1EE1) D642 ,"em. 4str.- 116(10)> D3662. B6EC P.*. +utaa- &.(.). Ravi *umar- Laste utili6ation> c"itosan 74res 4y #irect #issolution- in> 0n#ian ,"emist ,onvention(ew +el"i- +ec. 1720- 1EE7. B70C R. . . &u66arelli- ?uman en6ymatic activities relate# to t"e t"era$eutical a#ministration o! c"itin #erivatives- ,ell &ol. :iol. Ai!e =ci. 3G (1EE7) 1G1. B71C R.A. L"istler (<#.)- Polysacc"ari#e ,"emistry- ca#emic Press- (ew Sor/- 1EDG- $. GE3. B72C &. Sal$ani- F. 1o"nson- A.<. Ro4inson- ,"itin- ,"itosan> =ources- ,"emistry- :ioc"emistry- P"ysical Pro$erties an# $$lications- <lsevier- mster#am- 1EE2. B7GC <.R. Pariser- +.P. Aom4a#i- ,"itin =ource :oo/> Mui#e to Researc" Aiterature- Liley- (ew Sor/- 1ED0. B74C R. . &u66arelli- &. &attioli@:elmonte- . Pu5naloni- M. :ia5ini- :ioc"emistry- "istolo5y an# clinical uses o! c"itins an# c"itosans in woun# "ealin5- in> P. 1olles- R. . . &u66arelli (<#s.)- ,"itin an# ,"itinases- :ir/"auser- :asel1EEE. B73C &.(.). Ravi *umar- ,"itin an# c"itosan 74res> a review:ull. &ater. =ci. 22 (1EEE) E03.

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 B102C *.M.R. (air- P. &a#"avan- ,"itosan !or removal o! mer@ cury !rom water- Fis"ery 9ec". 21 (1ED4) 10E. B10GC ,. Penic"e@covas- A.L. lware6- L. r5uelles@&onal- 9"e a#sor$tion o! mercuric ions 4y c"itosan- 1. $$l. Polym. =ci. 46 (1ED7) 1147. B104C (. 1"a- 0. Aeela- .).=. Pra4"a/ar Rao- Removal o! ca#@ mium usin5 c"itosan- 1. <nviron. <n5. 114 (1EDD) E62. B103C &. Leltrows/i- :. &artel- &. (orcellet- 1. $$l. Polym. =ci. 3E (1EE6) 647. B106C +.L. *an5- ?.*. ,"oi- +.*. *weon- Polymer (*orea) 20 (1EE6) EDE. B107C M. &c*ay- ?.=. :lair- 1.R. Mar#ner- #sor$tion o! #yes on c"itin. 0. <8uili4rium stu#ies- 1. $$l. Polym. =ci. 27 (1EDE) G04G. B10DC 9.,. San5- R.R. Tall- 0&<, Pro#uct R&+ 2G (1ED4) 16D. B10EC R. &aruca- :.1. =u#er- 1.P. Li5"tmen- 1. $$l. Polym. =ci. 27 (1ED2) 4D27. B110C &.(.). Ravi *umar- 9. Ra'a/ala =ri#"ari- *. +ur5a :"avani- P.*. +utta- 9ren#s in color removal !rom te;tile mill e!Kuents- ,olora5e u5. (1EED) 23. B111C =. ?arry- 9"e t"eory o! coloration o! te;tiles- in> . 1o"nson (<#.)- 9"ermo#ynamics o! +ye =or$tion- 2n# <#ition- =ociety o! +yers an# ,olorists- Lest Sor/s"ire.*- 1EDE- $. 233. B112C L.:. Le4er- P"ysicoc"emical Process !or Lastewater ,on@ trol- Liley- (ew Sor/- 1EE2. B11GC F.L. =osuls/i- 9"e centri!u5ation met"o# !or #eterminin5 Kour a4sor$tion in "ar# re# s$rin5 w"eats- ,ereal ,"em. GE (1E62) G44. B114C P.*. +utta- &.(.). Ravi *umar- 9e;tile in#ustries> sa!ety"ealt" an# environment- in> R.*. 9rive#y (<#.)- #vances in Lastewater 9reatment 9ec"nolo5ies- Mlo4al =cience0n#ia- 1EED- $. 22E. B113C +. *norr- Functional $ro$erties o! c"itin an# c"itosan- 1. Foo# =ci. 47 (1ED2) 3EG. B116C +. *norr- +ye 4in#in5 $ro$erties o! c"itin an# c"itosan- 1. Foo# =ci. 4D (1EDG) G7. B117C M. llan- M.+. ,ros$y- 1.?. Aee- &.A. &iller- L.&. Rei!in> Procee#in5s o! a =ym$osium on &an@ma#e Polymers in Pa$er &a/in5- ?elsin/i- Finlan#- 1E72. B11DC A. ro!- R.?.S. =u44an- =. Ra#"a/ris"na- in> P.(. Prasa# (<#.)- Polymer an# Jt"er #vance# &aterials> <mer5in5 9ec"nolo5ies an# :usiness- Plenum Press- (ew Sor/- 1EE3$. 3GE. B11EC *.<. ."ric"- =.&. ,anni66aro- R.=. Aan5er- *.&. ="a/es"e!!- Polymeric systems !or controlle# #ru5 release,"em. Rev. EE (1EEE) G1D1. B120C ,. lvare6@Aoren6o- R. +uro- 1.A. Mome6@ mo6a- R. &artin@Pac"eco- ,. =outo- . ,onc"eiro- +e5ra#ation o! "y#ro;y$ro$ylcellulose 4y r"i6omucor> e!!ects on release !rom t"eo$"ylline"y#ro;y$ro$ylcellulose ta4lets- 0nt. 1. P"arm. 1D0 (1EEE) 103. B121C ,. lvare6@Aoren6o- 1.A. Mome6@ mo6a- R. &artin@Pac"@ eco- ,. =outo- . ,onc"eiro- &icroviscosity o! "y#ro;y@ $ro$ylcellulose 5els as a 4asis !or $re#iction o! #ru5 #i!!usion rates- 0nt. 1. P"arm. 1D0 (1EEE) E1. B122C +. 9eomim- . (ys/a- .1. +om4- Ricinoleic aci# 4ase# 4io$olymers- 1. :iome#. &ater. Res. 43 (1EEE) 23D. B12GC R.A. Reis- .&. ,un"a- ,"aracteri6ation o! two 4io#e5ra@ #a4le $olymers o! $otential a$$lications wit"in t"e 4ioma@ terial 7el#- 1. &ater. =ci.> &ater. &e#. 6 (1EE3) 7D6.

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B124C &.(.). Ravi *umar- M. &a#"ava Re##y- P.*. +utta- =tu#y o! $aracetamol release !rom castor@oil 4ase# co$olyester matri;- 0ranian Polym. 1. 3 (1EE6) 60. B123C &.(.). Ravi *umar- (. *umar- Polymeric controlle# #ru5 #elivery systems> $ers$ective issues an# o$$ortunities+ru5 +ev. 0n#. P"arm. (in $ress). B126C *.,. Mu$ta- &.(.). Ravi *umar- =tructural c"an5es an# release c"aracteristics o! crosslin/e# c"itosan 4ea#s in res$onse to solution $?- 1.&.=.@Pure $$l. ,"em. G6 (1EEE) D27. B127C *.,. Mu$ta- &.(.). Ravi *umar- +ru5 release 4e"avior o! 4ea#s an# micro5ranules o! c"itosan- :iomaterials 21 (2000) 1113. B12DC *.,. Mu$ta- &.(.). Ravi *umar- Pre$aration an# c"arac@ teri6ation an# release $ro7les o! $? sensitive c"itosan 4ea#s- Polym. 0nt. 4E (2000) 141. B12EC *.,. Mu$ta- &.(.). Ravi *umar- =emi@inter$enetratin5 $olymer networ/ 4ea#s o! crosslin/e# c"itosan5lycine !or controlle# release o! c"lor$"enaramine maleate- 1. $$l. Polym. =ci. 76 (2000) 672. B1G0C L. R"ine- +. ?sie"- R. Aan5er- Polymers o! sustaine# macromolecule release> $ro#ucers to !a4ricate re$ro#uci4le #elivery systems an# controlle# release /inetics- 1. P"arm. =ci. 6E (1ED0) 263. B1G1C &.(.). Ravi *umar- (ano an# micro$articles as controlle# #ru5 #elivery #evices- 1. P"arm. P"armaceut. =ci. (in $ress). B1G2C ). *umar- =.?. *ot"ari- <!!ect o! com$ressional !orce on t"e crystallinity o! #irectly com$ressi4le cellulose e;ci$i@ ents- 0nt. 1. P"arm. 177 (1EEE) 17G. B1GGC L.&. ?ou- =. &iya6a/i- &. 9a/a#a- 9. *omai- =ustaine# release o! in#omet"acin !rom c"itosan 5ranules- ,"em. P"arm. :ull. GG (1ED3) GED6. B1G4C =. &iya6a/i- *. 0s"ii- 9. (a#ai- 9"e use o! c"itin an# c"itosan as #ru5 carriers- ,"em. P"arm. :ull. 2E (1ED1) G067. B1G3C 9. ,"an#y- ,.P. ="arma- ,"itosan R as a 4iomaterial:iomater. rti!. ,ells rti!. Jr5ans 1D (1EE0) 1. B1G6C 9. ,"an#y- ,.P. ="arma- :io#e5ra#a4le c"itosan matri; !or t"e controlle# release o! steroi#s- :iomater. rti!. ,ells 0mmo4ili6ation :iotec"nol. 1E (1EE1) 743. B1G7C 1. *ost- R. Aan5er- in> (. . Pe$$as (<#.)- ?y#ro5els in &e#icine an# P"armacy- ,R, Press- :oca Raton- 1ED7- $. E3. B1GDC (.:. Mra"am- ,ontrolle# #ru5 #elivery systems- ,"em. 0n#. 13 (1EE0) 4D2. B1GEC *.+. Sao- 9. Pen5- ?.:. Fen5- S.S. ?e- =wellin5 /inetics an# release c"aracteristics o! crosslin/e# c"itosan$oly@ et"er $olymer networ/ (semi@0P() "y#ro5els- 1. Polym. =ci.- Part > Polym. ,"em. G2 (1EE4) 121G. B140C *.+. Sao- 9. Pen5- &.Q. Qu- ,. Suan- &.F. . Moosen- U. T"an5- A. Ren- $? #e$en#ent "y#rolysis an# #ru5 release o! c"itosan/$olyet"er inter$enetratin5 $olymer networ/ "y#ro5el- Polym. 0nt. G4 (1EE4) 21G. B141C 9. =a/iyama- ,.?. ,"u- 9. Fu'i- 9. Sano- Pre$aration o! $olyelectrolyte com$le; 5el !rom c"itosan an# /@car@ ra5eenan an# its $? sensitivity swellin5- 1. $$l. Polym. =ci. 30 (1EEG) 2021. B142C 1. :ronste#- 1. *o$ece/- ?y#ro5els !or site s$eci7c oral #ru5 #elivery synt"esis an# c"aracteri6ation- :iomaterials 12 (1EE1) 3D4.

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&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 B160C =. &iya6a/i- ,"itin an# c"itosan as ve"icle !or #ru5 #elivery- Tairyo Mi'utsu 16 (1EED) 276. B161C S. =awayana5i- (. (am4u- 9. (a5ai- ,om$resse# ta4lets containin5 c"itin an# c"itosan in a##ition to lactose or $otato starc"- ,"em. P"arm. :ull. G0 (1ED2) 2EG3. B162C M.,. Ritt"i#e'- P. ,"omto- =. Pumman5ura- P. &enasveta,"itin an# c"itosan as #isinti5rants in $aracetamol ta4lets+ru5 +ev. 0n#. P"arm. 20 (1EE4) 201E. B16GC =.0. Pat"er- 0. Russell- 1. . =yee- =.?. (eau- =ustaine# release t"eo$"ylline ta4lets 4y #irect com$ression. Part 0> Formulation an# in vitro testin5- 0nt. 1. P"arm. 164 (1EED) 1. B164C F.A. &i- (.A. ?er- ,.S. *aun- 9. Lon5- =. ="yu- ,"itosan ta4lets !or controlle# #ru5 release o! t"eo$"ylline> e!!ect o! $olymer #ru5 wet or #ry 4len#in5 an# anioniccationic inter $olymer- 1. $$l. Polym. =ci. 66 (1EE7) 24E3. B163C (. . Pe$$as- 1.1. =a"lin- sim$le e8uation !or t"e #escri$@ tion o! solute release. 000. ,ou$lin5 o! #i!!usion an# rela;ation- 0nt. 1. P"arm. 37 (1EDE) 16E. B166C P.A. Rit5er- (. . Pe$$as- sim$le e8uation !or #escri$tion o! solute release. 0. Fic/ian an# non@Fic/ian release !rom non@swella4le #evices in t"e !orm o! sla4s- s$"eres- cylin@ #ers or #iscs- 1. ,ontrol. Release 3 (1ED7) 2G. B167C *.+. Sao- 9. Pen5- 1.1. Su- &.Q. Qu- &.F. . Moosen&icroca$sules/micros$"eres relate# to c"itosan- 1.&.=.@ Rev. &acromol. ,"em. P"ys. ,G3 (1EE3) 133. B16DC S.1. Su'i- &.Q. Qu- Q. ,"en- *.+. Sao- +ru5 release 4e"avior o! c"itosan/5elatin networ/ $olymer micro@ s$"eres- ,"in. =ci. :ull. 41 (1EE6) 1266. B16EC &.,. Mo"el- &.(. ="et"- &.&. Patel- M.*. 1ani- ?. Patel+esi5n o! c"itosan micros$"eres containin5 #iclo!enac so#ium- 0n#ian 1. P"arm. =ci. 36 (1EE4) 210. B170C R. Mre!- S. &inamita/e- &.9. Peracc"ia- ). 9ru4ets/oy- ). 9orc"ilin- R. Aan5er- :io#e5ra#a4le lon5@circulatin5 $oly@ meric micros$"eres- =cience 1D (1EE4) 1600. B171C &. mi'i- *. Par/- in> =.L. ="ala4y- S. 0/a#a- R. Aan5er- 1. Lilliams (<#s.)- Polymers o! :iolo5ical =i5ni7cance- ,= =ym$osium =eries- )ol. 340- ,=- Las"in5ton- +,- 1EE4. B172C ,.&. Ae"r- 1. . :oustra- <.?. =c"ac"t- ?.<. 1un5in5er- 0n vitro evaluation o! mucoa#"esive $ro$erties o! c"itosan an# some ot"er natural $olymers- 0nt. 1. P"arm. 7D (1EE2) 4G. B17GC =. Pornsa/- +elivery systems !or $e$ti#e an# $rotein #ru5sLarasan P"esatc"asat 2G (1EE6) 1. B174C P. ,alvo- ,. Remunan@Ao$e6- 1.A. )ila@1ato- &.1. lonso(ovel "y#ro$"ilic c"itosan$oly et"ylene o;i#e nano$arti@ cles as $rotein carriers- 1. $$l. Polym. =ci. 6G (1EE7) 123. B173C &.A. ?u5uet- . Mro4oillot- R.1. (eu!el#- +. Poncelet- <. +ellac"erie- ?emo5lo4in enca$sulation in c"itosan/calcium al5inate 4ea#s- 1. $$l. Polym. =ci. 31 (1EE4) 1427. B176C &.A. ?u5uet- <. +ellac"erie- ,alcium al5inate 4ea#s coate# wit" c"itosan> e!!ect o! structure enca$sulate# materials on t"eir release- Process :ioc"em. G1 (1EE6) 743. B177C &.A. ?u5uet- R.1. (eu!el#- <. +ellac"erie- ,alcium al5i@ nate 4ea#s coate# wit" $olycationic $olymers> com$arison o! c"itosan an# +< <@#e;tran- Process :ioc"em. G1 (1EE6) G47. B17DC *. (is"imura- =. (is"imura- ?. =eo- (. (is"i- =. 9o/ura- 0. 6uma- &acro$"a5e activation wit" multi$orous 4ea#s $re$are# !rom $artially #eacetylate# c"itin- 1. :iome#. &ater. Res. 20 (1ED6) 1G3E.

B14GC =.:. &itra- in> =.L. ="ala4y- .=. ?o!!man- :.+. Ratner9. . ?or4ett (<#s.)- Polymers as :iomaterials- Plenum Press- (ew Sor/- 1ED4- $. 2EG. B144C 1. *ost (<#.)- Pulse# an# =el!@Re5ulate# +ru5 +elivery,R, Press- :oca Raton- 1EE0. B143C ,.P.+. &oor- A. +o"- R. . =ie5el- Aon5 term structural c"an5es in $?@sensitive "y#ro5els- :iomaterials 12 (1EE1) DG6. B146C . $ocella- :. ,a$$ello- &. . +elno4ile- &.0. Aaroton#aM. &ensitieri- A. (icolao- Poly(et"ylene o;i#e) P<J an# #i!!erent molecular wei5"t P<J 4len#s as monolit"ic #evices !or #ru5 release- :iomaterials 14 (1EEG) DG. B147C *.+. Sao- 9. Pen5- &.F. . Moosen- 1.&. &in- S.S. ?e- $? sensitivity o! "y#ro5els 4ase# on com$le; !ormin5 c"itosan> $olyet"er inter$enetratin5 $olymer networ/- 1. $$l. Polym. =ci. 4D (1EEG) G4G. B14DC 9. Pen5- *.+. Sao- &.F. . Moosen- =tructural c"an5es o! $? sensitive c"itosan/$olyet"er "y#ro5els in #i!!erent $? solution- 1. Polym. =ci.- Part > Polym. ,"em. G2 (1EE4) 3E1. B14EC *.+. Sao- 1. Ain- R.T. T"ao- L.?. Lan5- A. Lei- +ynamic water a4sor$tion c"aracteristics o! c"itosan 4ase# "y#ro@ 5els- n5ew. &a/romol. ,"em. 233 (1EED) 71. B130C =.=. *im- S.&. Aee- ,.=. ,"o- =ynt"esis an# $ro$erties o! semi@inter$enetratin5 $olymer networ/s com$ose# o! b@ c"itin an# $oly(et"ylene 5lycol) macromer- Polymer G6 (1EE3) 44E7. B131C =.=. *im- S.&. Aee- ,.=. ,"o- =emi@inter$enetratin5 $oly@ mer networ/s com$ose# o! b @c"itin an# $oly(et"ylene 5lycol) macromer- 1. Polym. =ci.- Part > Polym. ,"em. GG (1EE3) 22D3. B132C S.&. Aee- =.=. *im- ,.=. ,"o- Loun# coverin5 materials o! semi@inter$enetratin5 $olymer networ/ "y#ro5els com$ose# o! $oly(et"ylene 5lycol) #iacrylate macromers an# b @c"itin an# t"eir release o! silver sul!a#ia6ine- in> G6t" 0.P , 0nternational =ym$osium on &acromolecules- =eoul*orea- u5. 4E- 1EE6. B13GC S.&. Aee- =.=. *im- ?y#ro5els o! $oly(et"ylene 5lycol)@co@ $oly(lactones) #iacrylate macromers an# b @c"itin- Polymer GD (1EE7) 2413. B134C =.S. *im- S.&. Aee- =.0. Aee- Pre$aration an# evaluation o! in vitro sta4ility o! li$i# nanos$"eres containin5 vitamins or < !or cosmetic a$$lication- in> 24t" 0nternational =ym@ $osium on ,ontrolle# Release o! :ioactive &aterials=toc/"olm- =we#en- 1une 131E- 1EE7. B133C S.&. Aee- 1.*. ="im- Pre$aration o! $?/tem$erature stimuli res$onsive $olymer mem4ranes 4y $lasma an# .) $olymeri6ation tec"ni8ues an# t"eir ri4oKavin $ermeationin> 214t" ,= &eetin5- Aas )e5as- =e$t. 711- 1EE7. B136C S.&. Aee- =.=. *im- =.?. *im- =ynt"esis an# $ro$erties o! $oly(et"ylene 5lycol) macromer/ b @c"itosan "y#ro5els- 1. &ater. =ci.> &ater. &e#. D (1EE7) 3G7. B137C *.+. Sao- S.1. Sin- &.Q. Qu- F. Lan5- 0nvesti5ation o! $? sensitive #ru5 #elivery system o! c"itosan/5elatin "y4ri# $olymer networ/- Polym. 0nt. GD (1EE3) 77. B13DC ?. =e/i- 9. *a5ami- 9. ?ayas"i- (. J/usa- =ta4ility o! #ru5s in a8ueous solutions 00. $$lication o! Lei4ull $ro4a4ility $a$er to $re#iction o! coloration o! $aranteral solution- ,"em. P"arm. :ull. 2E (1ED1) G6D0. B13EC *. *o"ei- :iome#ical $olymers- Polymer 14 (1EE7) 22E.

materials- Polym. 0nt. 4D (1EEE) 7G2

&.(.). Ravi *umar / Reactive & Functional Polymers 46 (2000) 127 B17EC M. 0#a- ,. &onica- . nnalia- ,. :ice- &. Auisa- 0nKuence o! 5lutaral#e"y#e on #ru5 release an# mucoa#"esive $ro$@ erties o! c"itosan micros$"eres- ,ar4o"y#r. Polym. G6 (1EED) D1. B1D0C 9. ?ayas"i- S. 0/a#a- Protease immo4ili6ation onto $orous c"itosan 4ea#s- 1. $$l. Polym. =ci. 42 (1EE1) D3. B1D1C 9. ,"an#y- ,.P. ="arma- ,"itosan 4ea#s an# 5ranules !or oral sustaine# #elivery o! ni!e#i$ine> in vitro stu#ies:iomaterials 1G (1EE2) E4E. B1D2C 9. ,"an#y- ,.P. ="arma- ,"itosan matri; !or oral sustaine# #elivery o! am$icillin- :iomaterials 14 (1EEG) EGE. B1DGC 9. ,"an#y- ,.P. ="arma- &.,. =unny- 0n"i4ition o! $latelet a#"esion to 5low #isc"ar5e mo#i7e# sur!aces- 1. :iomater. $$l. 1 (1ED7) 3GG. B1D4C +. 9"ac"aro#i- *. Pan#uran5a Rao- Pro$ranolol "y#roc"lo@ ri#e release 4e"avior o! crosslin/e# c"itosan mem4rane- 1. ,"em. 9ec"nol. :iotec"nol. 3D (1EEG) 177. B1D3C +. 9"ac"aro#i- *. Pan#uran5a Rao- Release o! ni!e#i$ine t"rou5" crosslin/e# c"itosan mem4ranes- 0nt. 1. P"arm. E6 (1EEG) GG. B1D6C +. 9"ac"aro#i- *. Pan#uran5a Rao- +evelo$ment an# in vitro evaluation o! c"itosan 4ase# trans#ermal #ru5 #elivery systems !or controlle# #elivery o! $ro$ranolol "y#roc"lo@ ri#e- :iomaterials 16 (1EE3) 143. B1D7C =. ?irano- ,"itin an# c"itosan as novel 4iotec"nolo5ical

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B1DDC =. ?irano- &. ?ayas"i- =. J/uno- in> L.F. =tevens- &.=. Rao- =. ,"an#ra/ranc"an5 (<#s.)- ,"itin an# ,"itosan. <nvironmental Frien#ly an# )ersatile :iomaterials- 09:an5/o/- 1EE6- $. 1DD. B1DEC =. ?irano- in> ,.M. Me4elein- ,.<. ,arra"er1r. (<#s.)0n#ustrial :iotec"nolo5ical Polymers- 9ec"nomic- Aancas@ ter- 1EE3- $. 1DE. B1E0C =. ?irano- in> L.F. =tevens- &.=. Rao- =. ,"an#ra/ran@ c"an5 (<#s.)- ,"itin an# ,"itosan. <nvironmental Frien#ly an# )ersatile :iomaterials- 09- :an5/o/- 1EE6- $. 22. B1E1C 1. La#stein- <. 9"om- <. ?el#man- =. Mu#munsson- :. Ail'a- :io$olymer A 112- a c"itosan wit" !at 4in#in5 $ro$erties an# $otential as a wei5"t re#ucin5 a5ent> a review o! in vitro an# in vivo e;$eriments- in> R. . . &u66arelli (<#.)- ,"itosan Per os> From +ietary =u$$le@ ment 9o +ru5 ,arrier- Mrottammare- 0taly- 2000. B1E2C R. . . &u66arelli- &ana5ement o! "y$erc"olesterolemia an# overwei5"t 4y oral a#ministration o! c"itosans- in> +. ,"a$man- P.0. ?aris (<#s.)- (ew :iome#ical &aterials R $$lie# an# :asics- PJ0 Press- Aon#on- 1EED. B1EGC R. . &u66arelli- ,linical an# 4ioc"emical evolution o! c"itosan !or "y$erc"olesterolemia an# overwei5"t controlin> P. 1olles- R. . . &u66arelli (<#s.)- ,"itin an# ,"itin@ ases- :ir/"auser- :asel- 1EEE.