Step By Experimental

1. Sample Preparation

Shrimp shell waste is cleaned of residual dirt and shrimp meat is left on the

skin. Shrimp shells washed with clean running water for 3 times, the washing process

shrimp shells to obtain a clean and odorless. Dried shrimp shells under the blazing

sun for hours with 2x24 covered with black cloth to prevent direct exposure to the

sun can damage the sample. After drying, and then blended to a powder to expand the

sample surface and subsequently sieved and the results floured shrimp shells are used

as raw materials in this study.

2. Isolation of Chitin

Chitin isolation method performed according Hang. Chitin isolation includes

the step of demineralization and decolorization deproteinasi or depigmentation

(Purwatiningsih, 1993).

a. Demineralization

As much as 50 grams of powder shell of shrimp and then dissolved in a

solution of hydrochloric acid (HCl) 1.0 M in comparison to 1:10 (sample:

solvent), then stirred with stirrer for 1 hour at a temperature of 75oC. Further

filtered using the filter paper and residue produced washed by using aquades

to pH neutral, then dried in an oven at the temperature 80oC.

 b. Deproteinization

Sample result demineralization then deproteinization using NaOH 3.5%

in comparison with the 1:10 (sample: solvent), then stirred with stirrer for 2

hours at a temperature of 65oC. Then the sample is filtered and washed with

aquades to neutral pH and dried in the oven at the temperature 80oC.

c. Decolorization

The result of the stage (a) and (b) further weighed and dissolved into

0.315% NaOCl by comparison 1:10 (sample: solvent), then put in the

erlenmeyer flask and stirred for 5 minutes at room temperature. Further

filtered using the filter paper and residue produced washed by using aquades

to pH neutral, then dried in an oven at the temperature 80oC.

3. Making Of Glucosamine

Production of Glucosamine hydrochloride starts with soaking chitin as much

as 3 grams of hydrochloric acid in aqueous solution with a concentration of 37%

(Mojarrad et al., 2007) and is carried out at a temperature 90oC (Ernawati, 2012) then

enter into a microwave with a reaction that is old, 7.11 3 and 15 minutes.

4. Characterization of Glucosamine

a. Degree Deacetylation of chitin with FTIR (Fourier Transform-Infra Red)

(Azhar.2010).
 The degree of deacetilation chitin is determined by baseline methods. This

method is based on comparison of the values of absorbance of the

spectrum of infrared absorption bands in the wave number 1655 cm-1 and

3450 cm-1. Absorbance (A) is expressed as equation (1), while the value of

DD is expressed as equation (2).

The value of absorbans can be measured by using the formula:

𝑃𝑂
𝐴 = 𝐿𝑜𝑔 … … … … … … … … … … … … … . . (1)
𝑃

Where :

A: Absorbans

PO : % transmitan on base-line

P : % transmitan on maxsimum absorption

𝐴1656 1
% 𝐷𝐷 = (1 − ( × )) × 100% … . (2)
𝐴1343 1,33

A1655 and A3450 is A value that corresponds to the tape 1655 cm-1

absorption and 3450 cm-1. 1655 cm –1 absorption bands is Carbonyl

absorption bands of functional N-acetyl 3450 cm-1 absorption bands while

the absorption bands cluster NH2.

b. Analysis of the structure of Crystalline Glucosamine use XRD.

c. Glucosamine Molecular Weight Analysis using Ostwald viscometer.