Saran Pengembangan kitosan sebagai absorben pengotor dalam aplikasi pemurnian agar dan karagenan
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LAMPIRAN
Lampiran 1 Tabel US Paten mengenai penggunaan kitosan sebagai absorben
No No patent Inventor
Tahun Judul patent 1
4,895,724 Cardinal;John R. Old Lyme,
CT; Curatolo; William J. Old Lyme, CT; Ebert; Charumput
lautesBrewster, NY 1985
Chitosan composition for
controlled and prolonged release
of macromolecules
2 6,800,789
Kasai; Takao Haga-gun, JP; Kondo; Megumi Haga-gun,
JP; Sato; Noriko Haga-gun, JP; Matsui; Manabu Haga-
gun, JP 2001
Absorbent article
3 6,786,336
Boddu;Veera M. Champaign, IL; Smith;Edgar Dean
Seymour, IL 2003
Composite biosorbent for
treatment for waste aqueous systems
containing heavy metals
4 6,753,179
Tsuciya; Akihito Shiga-ken, JP
2000 Method for
purification treatment of
environmental pollutants
5 5,993,668
Duan; Hailing Allendele, NJ 1997
Methode for removing metal
ions andor complexes
containing metal ions for a solution
6 6,465,521
Rosenberg; Melvyt Ramat- Gan, IL
1994 Composition for
desorbing bacteria 7
6,599,290 Nakatani; MasaruKobe, JP;
Furuyoshi; ShigeoKobe, JP; Takata; SatoshiTakasgo, JP
1997 Methode for
removing a chemokine
8 5,516,569
Veith;Michael W.Oshkosh,WI;Abuto;Francis
P.Alpharetta,GA;Werner;Edw ard
E.Oshkosh,WI;Wisneski;Anth ony J.Kimberumput lauty,WI
1993 High absorbency
composite
9 5,750,065
Kilbane,II;John J. Woodstock,IL
1993 Adsorption of
PCB’s using biosorbents
Lampiran 1 lanjutan Lanjutan tabel US Paten mengenai penggunaan kitosan sebagai absorben
10 5,789,204
Kogtev;Leonid Semionovich Moscow,RU;Park;Jin Kyu
Seoul,KR;Pyo;Jin Kyuk Seoul,KR;Mo;Young Keun
Chungbook,KR 1996
Biosorbent for heavy metals
prepared from biomass
11 4,651,725
Kifune;Koji Nara,JP;Yamaguchi;Yasuhiko
Kyoto,JPTanae;Hiroyuki Kyoto,JP
1986 Wound dressing
12 5,051,293
Mayer;Jean M. Smithfield,RI;Kaplan;David
L. Stow,MA 1989
Method of forming a crosslinked
chitosan polymer and product thereof
13 6,833,487
Pesce; Antonella Pescara, IT; Tordone; Adelia Alessandra
Pescara, IT; Carumput lautucci; Giovanni Chieti,IT;
Di Cintio; Achille Pescara,IT 2002
Articles compirising a
cationic polysaccharide and
silica
14 6,844,430
Pesce;Antonella Pescara,IT;Tordone; Adelia
Alessandra Pescara,IT; Carumput lautucci; Giovanni
Chieti,IT; Di Cintio; Achille Pescara,IT
2002 Articles
compirising cationic
polysaccharides and acidic pH
buffering means
15 5,110,733
Kim; Chan W. Cambridge, MA; Robinson; Elizabeth M.
Cambridge, MA;Rha; Chokyun Boston, MA
1989 Liquid-liquid
extraction with particulate polymer
adsorbent
16 4,971,698
Weber; Alfred Berumput lautin, DE; Klages; Uwe
Berumput lautin, DE; Donner; Cristoph Berumput lautin, DE
1989 Process for
wastewater puritfication
17 4,424,346
Hall; Laurance D. Vancouver, Ca; Yalpani; Mansur
Vancouver, CA 1981
Derivative of chitins, chitosans
and other polysaccharides
Lampiran 2 Hasil analisis karotenoid terabsorbsi
Konsentrasi kitosan
Absorbansi Rata-rata
Standar deviasi Konsentrasi
karotenoid
0,1 0,0860
0,0861 0,0001
±9,11 mg 0,0862
0,2 0,9770
0,0972 0,0085
±10,2 mg 0,9650
0,3 0,1224
0,1224 0,0000
±12,5 mg 0,1224
0,4 0,1401
0,1402 0,0001
±17,4 mg 0,1403
0,5 0,1639
0,1642 0,0004
±20,1 mg 0,1645
0.02 0.04
0.06 0.08
0.1 0.12
0.14 0.16
0.18
0.1 0.2
0.3 0.4
0.5
Konsentrasi Kitosan A
b s
o r
b s
i E
k s
tr a
k W
o r
te l
C karotin mg
Absorben 10
0,0925 20
0,1592 30
0,2130 40
0,2644
0.861±0.0001 0.0972±0.0085
0.1224±0.000 0.1402±0.0001
0.1642±0.0004
Lampiran 3 Komposisi Kimia Buffer Pospat Metoda Pembuatan Buffer Posfat pH7
Buffer Posfat dibuat melalui pencampuran larutan A dan Larutan B, masing- masing larutan terdiri dari komposisi sebgai berikut :
Larutan A Penimbangan : Na
2
HPO
4
2H
2
O sejumlah 17.799 gram dimasukan kedalam labu takar 1 liter dan dilarutkan dengan akuades serta dihomogenkan kemudian
dijadikan 1 liter. Larutan B
Penimbangan : NaH
2
PO
4
2H
2
O sejumlah 15.601 gram dimasukan kedalam labu takar 1 liter dan dilarutkan dengan akuades serta dihomogenkan kemudian
dijadikan 1 liter. Campurkan 61.1 ml larutan A dengan 38.9 ml larutan B, menjadi 100 ml buffer
posfat pH 7 yang siap digunakan.
Lampiran 4 Gambar alat SEM dan AAS.
Lampiran 5 Spektrogram FTIR dari kitosan terpilih
Lampiran 6 Spektrogram FTIR dari kitosan komersil.
Lampiran 7 Hasil analisis Autosorp.
Lampiran 8 Hasil Analisis Spektrofotometer dari FeSO
4
a. Tabel standar FeSO
4
b. Tabel dan kurva absorbsi FeSO
4
Lampiran 9 Hasil Analisis Spektrofotometer dari CuSO
4
a. Tabel dan kurva standar CuSO
4
b. Tabel dan histogram absorbsi CuSO
4
Lampiran 10 Hasil Analisis Spektrofotometer dari Pb asetat b Tabel dan kurva baku Pb asetat
b. Histogram absorbsi Pb Asetat
Lampiran 11 Hasil Analisis Spektrofotometer dari E coli a Tabel dan kurva standar E coli
b Histogram absorbsi E coli oleh kitosan Konsentrasi E coli Absorbansi
0,10 1,10
0,20 1,45
0,30 1,52
0,40 2,11
0,50 2,48
Konsentrasi kitosan
Absorbansi 0,05
0,85 0,10
0,96 0,20
0,10 0,30
0,11 0,40
0,12 0,50
0,12
Lampiran 12 Struktur karagenan dan typical bend dari karagenan
Lampiran 4. Kromatogram HPLC dari karagenan
Lampiran 13 Gambar spektrum HPLC dari karagenan. Alexander 1993
Lampiran 14 Diagram alir proses produksi kitosan dengan sistem zero waste
Kulit udang
Pencucian
Ekstraksi Ekstrak FLAVOUR
Demineralisasi HCl 1 N, 90
C, 1 Jam
KALSIUM Presifitasi
Netralisasi
Deproteinisasi NaOH 3 N, 90
C, 1 Jam
Netralisasi
KITIN
Deasetilasi NaOH 50, 140
C, 1 Jam
KITOSAN PROTEIN Presifitasi
KLOROASETAT
Larutan NaCl
HIPOKLORIT dengan
Elektrolisis A
A
Lampiran 15 Diagram alir proses produksi karagenan Brian 2000
PIPIH SUPTIJAH. C 526014011. Pengembangan Kitosan sebagai Absorben Pengotor dalam Aplikasi Pemurnian Agar dan Karagenan. Dibimbing oleh:
LINAWATI HARDJITO. JOHN HALUAN dan MAGGY T SUHARTONO.
RINGKASAN
Kitosan adalah polimer glukosamin yang sangat banyak dialam setelah selulosa. Sebagai polimer alami kitosan mempunyai muatan ionik yang reaktif
sehingga dapat mengikat dan mengabsorbsi komponen lain yang bermuatan berlawanan, oleh karena itu kitosan mempunyai kemampuan sebagai absorben.
Kitosan dimanfaatkan sebagai absorben terhadap pengotor dalam proses pemurnian rumput laut sehingga dihasilkan produk agar dan karagenan yang
bermutu baik.
Tujuan dari penelitian ini adalah: 1 Menentukan karakteristik fisika, kimia dan mikroskopis kitosan yang akan dikembangkan sebagai absorben 2
Menguji kemampuan kitosan sebagai absorben logam berat Fe Cu, Pb, pigmen
ekstrak wortel dan bakteri Escherichia coli E. coli 3 Mengaplikasikan kitosan sebagai absorben terhadap pengotor pada ekstraksi agar dan karagenan 4
Menganalisis mutu agar dan karagenan yang dihasilkan dari metoda tersebut.
Penelitian ini terdiri dari beberapa tahapan yang meliputi: tahap preparasi, tahap karakterisasi, dan tahap aplikasi. Tahap preparasi yaitu tahap produksi
kitosan dengan memodifikasi kondisi proses, melalui 36 perlakuan diantaranya variasi larutan NaOH 0,5-2N dan waktu proses deproteinisasi 2-5 jam serta
deasetilasi dengan NaOH 1,5-6N dilanjutkan dengan analisis mutu hasilnya. Tahap karakterisasi meliputi penentuan sifat fisika ukuran dan viskositas, kimia
proksimat dan derajat deasetilasi dan mikroskopis morfologi permukaan-pori- pori dari kitosan serta uji kemampuan absorbsinya terhadap logam Fe, Cu, Pb,
pigmen ekstrak wortel, dan bakteri E. coli. Tahap aplikasi meliputi tahap penggunaan absorben pada ekstraksi agar dan karagenan serta uji mutu hasil
aplikasinya dengan viskometer viskositas, rheoteks gel strength, FTIR gugus fungsi, SEM morfologi, autosorp distribusi pori, ONPG: Orto Nitro Phenil
Glikosida bakteri dan HPLC untuk uji komponen pengotor yang tersisa dan komponen utama -galaktosa yang sudah bersih.
Hasil kitosan yang dibuat dengan kondisi deproteinisasi: NaOH 1N, waktu proses 4jam, dan deasetilasi: NaOH 6N, waktu 2jam, terpilih sebagai absorben,
adalah kitosan dengan derajat deasetilasi 90, mempunyai karakteristik fisika- kimia sebagai berikut: rendemen 13,5, tidak berwarna, lebih transparan, kadar
N 4, kadar mineral 0,2, kadar air 10, viskositas 247 cPs, gugus fungsi amin terdeteksi oleh FTIR pada bilangan gelombang 1639 cm
-1
dan gugus fungsi hidroksi pada bilangan gelombang 3410 cm
-1
. Analisis SEM menunjukan morfologi kitosan yang ber pori- pori dan melalui analisis autosorp menunjukkan
distribusi pori dengan variasi diameter pori antara 37,2 Å sampai 1802205 Å. Kitosan 0,1 mempunyai kemampuan mengabsobsi 1 larutan logam Fe
32, logam Cu 26, logam Pb 22, ekstrak wortel sebanyak 50 100 bv pewarna minuman bersoda 55 dan 2 bv biomas E. coli 80,58.
Hasil aplikasi kitosan pada ekstraksi agar menunjukkan bahwa penambahan 0,1 kitosan sebagai absorben menghasilkan kualitas agar dengan
pembanding bakto agar sebagai standar adalah sebagai berikut: Total Plate Count 1,25 x10
2
CFU bakto agar 2,04 x10
2
CFU, kadar sulfat 0,1 bakto agar 0,3, kadar air 21,1 bakto agar 16,9 , kekuatan gel 275,10 gF bakto agar
350,15 gF. Hasil aplikasi kitosan pada ekstraksi karagenan menunjukkan kualitas
karagenan sebagai produk: kadar sulfat 12,40, kekuatan gel 80gF, kadar air15 dan viskositas 26,4cPs. Hasil analisis FTIR pada karagenan mengkonfirmasi tujuh
gugus fungsi OH terdeteksi pada 3000-3450cm
-1
, CH pada 2920 cm
-1
, amida pada 1650 cm
-1
, sulfat ester pada 1350-1355 cm
-1
, glikosidik pada 1150 cm
-1
, 3.6 anhidro galaktan pada 930 cm
-1
dan C2-O-S dalam galaktan pada 830 cm
-1
, sementara gugus fungsi sulfat terdeteksi pada bilangan gelombang 1350 cm
-1
. Hasil analisis HPLC karagenan menunjukkan karakteristik serapan paling bersih
pada penambahan kitosan 0,1. Dengan demikian penggunaan kitosan 0,1 baik diaplikasikan sebagai absorben dalam pemurnian agar dan karagenan.
Kata kunci: kitosan, absorben, pengotor, ekstrak wortel, ONPG.
PIPIH SUPTIJAH. C 526014011. Development of Chitosan as Impurity Absorbent on Agar and Carrageenan Purification
. Supervised by LINAWATI HARDJITO.
JOHN HALUAN and MAGGY T SUHARTONO. ABSTRACT
Chitosan, a polymer of glucosamine is the largest polysaccharides after cellulose. A natural polymer chitosan has reactive ionic charge that has ability to bind and
absorb other components of opposite charge as an absorbent. This research utilized chitosan in the process of seaweed purification .
The purpose of this study were 1 To determine physical, chemical and microscopic characteristics of chitosan that will be developed as an absorbent 2 To
test the ability of chitosan as an absorbent of heavy metals Fe, Cu, Pb, pigments carrot extract and bacteria E. coli 3 To apply chitosan to absorb impurity in the
extraction of agar and carrageenan 4 To analyze the quality of agar and carrageenan produced from these methods.
This study consisted of several stages which included: preparation, characterization and application phase. Preparation of chitosan production was
conducted by modifying the process conditions, through 36 variations treatment NaOH concentrations 0.5-2N and deproteinisation time 2-5 hours and
deacetylation with NaOH 1.5-6N, followed by quality analysis. The characterization phase involved determining the physical properties size and viscosity, chemical
proximate and degree of deacetylation and microscopic morphology-surface pores of chitosan.In addition the chitosan was tested to absorp metal Fe, Cu, Pb, pigments
carrot extract , and bacteria Escherichia coli. Stage of the application included the use of chitosan product on agar and carrageenan extraction, the product was
analysed using viscometer for viscosity, rheotex for gel strength, FTIR for functional group, SEM for morphology, Autosorp for pore distribution, ONPG: Ortho Nitro
Phenil glycosides for bacteria and HPLC for analysis of the remaining components of impurities and main compone
nts -galactose. The selected results were obtained using chitosan with deacetylation degree 90,which was made by using NaOH 1N
for 4 hours of deproteinization and deacetylation time for 2 hours with 6N NaOH. The chitosan showed a physical-chemical characteristics as follows: 13.5 yield,
colorless, transparent, N concentration of 4, 0.2 mineral content, water content 10, viscosity 247cPs at 1.5, have amine functional groups detected at 1639cm-1
and hydroxy group at 3410 cm-1. SEM analysis showed the morphology of chitosan- containing pores and through autosorp analysis showed the distribution of pores with
pore diameter variation between 37.2 Å up to 1,802,205 Å
0.1 chitosan has the ability to absorb 1 solution of Fe 32, Cu 26, Pb 22, carrot extract 100 wv by 50, soft drinks pigment 55 and 2 wv
biomass of E. coli by 80.58. 0.1 chitosan treatment for agar extraction as absorbent resulted agar quality bacto agar as a comparison as follows:
TPC 1.25
x 10
2
CFU 2.04
x10
2
CFU of bacto agar, 0.1 sulphate content 0.3 of bacto agar, water content 21.1 16, 9 of bacto agar, gel strength of 275.10 gF
350.15gF of bakto agar. The application to the extraction of carrageenan resulted : 12.40 sulphate content, gel strength of 80gF, 15 water concentration and viscosity
of 26.4 cps. FTIR analysis showed seven functional groups OH at 3000-3450 cm
-1
, CH at 2920 cm
-
1, amide at 1650 cm
-1
, sulphate ester at 1350-1355 cm
-1
, glycosidic bond at 1150 cm
-1
, 3.6 anhidro galaktan at 930 cm
-1
and C2-OS and galaktan at
830 cm
-1
, while the sulphate functional groups was detected only at 1350 cm-1. The result of HPLC analysis showed the clear pick at 0.1 chitosan treatment. Thus the
use of 0.1 chitosan was selected in the purification of agar and carrageenan as it is cheaper, simpler and non-chemical.
Key words: chitosan, absorbent, impurity, carrot extract, ONPG.
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1 PENDAHULUAN