Poster code In vitro Antioxidant activity of Chitin from Mud crab Scylla Serrate shells by

D-160
DPPH method

* 1
S.Ravindrababu ,Gudibandi Phani C Reddy
Department of pharmacology,Mallareddy institute of pharmaceutical sciences,
Dhullapally,Hyderabad.
Email id:- ravicology@gmail.com
ABSTRACT:
Chitin, a homopolymer of N-acetyl-D-glucosamine units linked by β-1-4 bonds, is the most widespread available natural resource of most of
marine byproducts obtained from crab shells of Scylla Serrate. They are known to possess Anti-oxidant activity by their natural habitat of living in marine environment, as
their shells are resistant to large number of micro-organisms of water from degradation. So, their resistance to shell degradation was thought to be due to the Anti-oxidant
effect which was assessed by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) method. The chitin from shells of Scylla Serrate as an alternative to chemical method was extracted by
biological method by using deproteination bacteria like P.aeruginosa K-187 and demineralization by using Alcalase enzyme in the process of fermentation. This method had
many advantages over chemical method as it is natural and less deleterious effects and economical and high yield. The chitin at different concentrations (20-100µg/ml) was
measured for percentage inhibitory effect (DPPH scavenging activity) against the standard drug L-Ascorbic acid by measuring absorbance at 517nm in spectrophotometer.
The results obtained shown % inhibitory activity of (56.6 ± 0.6) % for chitin at 100µg/ml concentration which was compared to standard L-Ascorbic acid (60.3 ± 0.2). Our
data shows that chitin obtained from shells of Scylla Serrate by biological method possess Anti-oxidant effect. 
Keywords :
DPPH, Scylla Serrate, P.aeruginosa K187, Chitin.

Introduction Antioxidant activity Figure-1

Chitin, a homopolymer of N-acetyl-D-glucosamine units DPPH radical scavenging activity

linked by β-1-4 bonds, is the most widespread available
natural resource of most of marine byproducts obtained
from crab shells of Scylla Serrate. Their shells are resistant
to large number of micro-organisms of water from
degradation which was due to their Anti-oxidant activity.
The free radicals are responsible for oxidation in many
cases so, it can be effectively assessed by DPPH method
which is based on free-radical scavenging activity. Chitin
extracted from mud crabs shells of Scylla Serrate are
thought to possess Anti-oxidant activity due to their marine
habitat.
The free-radical scavenging capacity of chitin was evaluated
with the DPPH stable radical. An 0.1mM alcoholic solution
of DPPH in methanol was prepared and 2mL of this solution
was added to 0.3mL of different chitin concentrations (20-
100 µg/ml) and allowed to react at room temperature. After
30min, the absorbance values were measured at 517nm
against the blank and L-Ascorbic acid as control.
The radical scavenging activity (AA %) was expressed as p
ercentage of DPPH radical elimination calculated according
to the following equation.

Values are Expressed as Mean ± S.D, n=3

(ABSsample – ABSblank) × 100
AA% = 100 - ABScontrol

Methodology
  Results Conclusion
Extraction of Chitin from Crab Shells from the results obtained the
The free radical scavenging activity can be assessed chitin extracted from shells of Scylla Serrate
  effectively by DPPH method which were shown in Table-1 possess Anti-oxidant activity as % inhibitory
and Figure-1. activity of (56.6 ± 0.6) % for chitin at 100µg/ml
Table-1 concentration which was compared to standard
L-Ascorbic acid (60.3 ± 0.2) where significant.
% of Scavenging activity
S.no (AA %)
Bibliography
Conc. µg/ml Ascorbic acid Chitin  
Hirano S,. Chitin biotechnology application.
1. 20 16.5±0.9 11.1±0.2 Biotechnol. Annu. (2006): 237-258.
 
2. 40 28.6±0.4 20.2±0.4 Chu Y, Flavonoid content of several vegetables
3. 60 46.5±0.6 40.4±0.3 and their antioxidant activity, J. Sci. Food and
Agriculture, (2000): 561-566.
4. 80 54.6±0.4 48.5±0.4
5. 100 60.3±0.2 55.6±0.6

Presented at the 66 Indian Pharmaceutical Congress held at Hitex, Hyderabad during 23 - 25 ,January 2015
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