Extraction Chitosan from Prawn Shells And its adsorption Studies on copper

1. EXTRACTION OF CHITOSAN FROM PRAWN
SHELLS AND OPTIMIZATION OF ITS ADSORPTION
PARAMETERS FOR THE REMOVAL OF COPPER
under the guidance of
Ms.P.MALLIKA RANI
Assistant professor
PRESENTED BY
Mr.S.MANIKANTA (316126502044)
Ms.S.KRISHNA VENI (316126502026)
Mr.N.MANOJ YADAV (316126502019)
Mr.M.VINEEL KUMAR (316126502033)

2. Contents
 Objective
 Properties of Chitin and Chitosan
 Applications
 Material and Methods
 Apparatus and Equipment
 Collection and washing of prawn shells
 Experimental procedure
 Visible absorption spectra of obtained chitosan
 Adsorption
 Selection of metal ion
 Study of Parameters
 Conclusion
 Future work

3. Objective
During the recent years waste has become a
major concern.
In this project the main aim is at reducing the
water pollution using the prawn shells wastage
as raw material for the extraction of chitin and
chitosan which is useful as adsorbent in waste
water treatment.

4. Structure of Chitosan

5. Chemical properties of Chitosan
Linear polyamine
Reactive amino groups
Reactive hydroxyl groups
Chelates many transitional metal ions

6. Linear polyamine
Reactive amino groups
Reactive hydroxyl groups
Chelates many transitional metal ions.
Chemical properties of Chitosan

7. Bio Compatable
Bio Degradable
Non-Toxic
Highly Hydro-Phobic
Anti -Tumour
Anti Cholesteremic
Accelerates Bone Formation.
Biological properties of Chitosan

8. Waste water treatment
Cosmetics
Paper industry
Textile industry
Photography
Pharmaceuticals
Wound healing
Applications

9. So extracted chitosan is used as flocculating
agent but in our study this chitosan is used as
adsorbent for the treatment of waste water
preferably removal of dyes and heavy metal ions.
Waste Water Treatment

10. There are 2 methods to extract chitin from
prawn shells:
1)Chemical
2)Biological
Methods

11. The following steps are to be followed in this
method :
1. De-mineralization
2. De-proteinization
3. De-colouration
4. De-acetylation
Chemical method

12. De-Mineralization
De-Proteinization
De-Colouration
De-Acetylation
HCl
NaOH
Acetone
Deactylation Reagent
HCl+ Calcium Carbonate
Protein+NaOH
Colors+Acetone
Chitosan
BLOCK DIAGRAM

13.  Chemicals required:
1)Hydrochloric acid(HCl)
2)Sodium hydroxide(NaOH)
3)Acetone
4)Deacetylation reagent
Materials

14. • Conical flask(250 ml)
• Beakers (400ml)
• Glass rod
• Measuring jar
• Funnel
• Magnetic Stirrer
• Heat source
Apparatus and Equipment

15. • Collected prawn shells waste from our local
markets.
• Dried it for 2-3 days and then removed the
unwanted residue(i.e., head and tails) from the
shells.
Collection and Washing of prawn
shells

16. • Shells are washed with water 2-3 times for
removal of salt and unwanted matter present on
the shells.
• Then taken in a beaker of 300 ml with water and
boiled at temperature of 80oC for about 45
minutes.
• Then the shells are filtered using mesh and the
obtained shells are dried for 3-4 hours and
grinded to fine powder.
Cleaning

18. De-mineralization
• The main inorganic component to be removed in
this process is calcium carbonate using HCl.
• The solution is taken into a beaker and the
prawn shell powder of 200g is added to HCl(1M)
solution in the ratio of 1:10(w/v).
• Then the solution is stirred with a magnetic
stirrer at a temperature of 400C for about 50
minutes.

19. Experimental Setup

20. • The solution is filtered using filter paper.
• The slurry is washed with distilled water and the
mixture is filtered with filter medium and the process is
repeated for 2 times.
• The obtained shells are then dried in oven at 80oC for
about 1 hour.
Filtration, Washing and Drying

21. De-proteinization
• After de-mineralisation process 131g of powder
was obtained.
• As per 1:10(w/v) ratio NaOH of 2M is required to
perform de-proteinization.
• The process is maintained at a temperature of
450C for about 6 hours.

22. Experimental Setup
during the process

23. • The solution is filtered using filter paper.
• The slurry is washed with distilled water and the
mixture is filtered with filter medium and the
process is repeated for 2 times.
• The obtained shells are then dried in oven at
800C for about 2 hours.
Filtration, Washing and Drying:

24. De-colouration
• After de-proteinization process, 67g of the
processed powder is obtained.
• For decolouration, acetone is used and it is
mixed with the obtained powder in the ratio of
1:10(w/v).
• Then the solution is stirred at room temperature
for about 10 minutes.

25. • The solution is filtered using filter paper.
• The slurry is washed with distilled water and the
mixture is filtered with filter medium and the
process is repeated for 2 times.
• The obtained shells are then dried in open
atmosphere at room temperature for a
overnight.
Filtration, Washing and Drying:

26. • Chitin will remain in the powder form after
decolouration process.

27. De-acetylation
• De-acetylation reagent is prepared using a
suspension of 1g of chitin in 50ml of NaOH.
• Chitin powder is dissolved with reagent in the
ratio of 1:10(w/v).
• The solution is stirred at a temperature of
1000C for about 4 hours.
• After stirring, the solid powder was filtered
and washed with water and 80% alcohol until
the filtrate was neutral and dried for
overnight at 800C.

28. Percentage of extraction
• 55.5g of chitosan is obtained from 200g of feed
% Extraction =55.5/200=0.2775=27.75 %
• The obtained chitosan is analysed by visible
spectrometer analysis.

29. Visible absorbtion spectra of obtained
chitosan
S.NO Wavelen
gth(nm)
Absorb
ance
1 354 0.275
2 370 0.25
3 390 0.225
4 480 0.172
5 550 0.134
6 650 0.114
7 750 0.106
8 850 0.09

30. Visible absorbtion spectra of pure
chitosan

31. Adsorption
• It is a process by which a solid holds molecules of a gas or
liquid as a thin layer on the surface of the solid.
• Types of adsorption are:
1. Physical
2. Chemical

32. Selection of metal ion
• Present study involves removal of copper metal ions.
• According to the United State Environmental Protection
Agency permissible limits of copper is 1.3mg/l in drinking
water and 1.9mg/l in industrial waste water.
• The obtained chitosan is used as an adsorbent for
wastewater treatment(by adsorption process).

33. Parameter studies
By varying
1) Dosage of adsorbent
2) Agitation time
3) pH of solution
4) Temperature of the solution
5) Size of the adsorbent

34. By varying dosage of adsorbent
• Firstly different dosage of adsorbent are taken
ranging from 0.5 to 3g.
• Copper solution of 0.001N is prepared as a
stock solution and 50 ml is taken in every
sample.

36. • All the parameters are kept constant such as:
• pH = 9.3
• Conc. of CuSO4 = 0.001 N
• Time = 60 min
• Temperature = 27˚C(room temp.)
• These samples are then kept in shaker for about
60 min and after completion the samples are
analyzed using visible spectrophotometer.

37. S.NO Dosage
(g)
%Adsorption
1 0.5 29.5
2 1.0 39.2
3 1.5 40.8
4 2.0 64.1
5 2.5 64.1
6 3.0 64.1

38. By varying the agitation time
• From the first parameter study it was observed
that dosage of chitosan is optimum at 2 g.
• By varying the agitation time ranging from 10-70
minutes.

39. • All the parameters are kept constant such as:
• pH = 9.3
• Conc. of CuSO4 = 0.001 N
• Dosage of chitosan = 2 g
• Temperature = 27 ˚C(room temp.)
• For every 10 min the samples are analyzed using
visible spectrophotometer.

40. S.NO Agitation
time(min)
%Adsorption
1 10 19.16
2 20 29.16
3 30 40
4 40 45.83
5 50 54.16
6 60 64.1
7 70 64.1

41. By varying the pH of solution
• From the above two parameter studies it was
observed that dosage of chitosan and agitation
time are optimum at 2 g and 60 minutes.
• By varying the pH of solution ranging from 4-10.

42. • All the parameters are kept constant such as:
• Time = 60 minutes
• Conc. of CuSO4 = 0.001 N
• Dosage of chitosan = 2 g
• Temperature = 27 ˚C(room temp.)
• These samples are then kept in shaker for about
60 min and after completion the samples are
analyzed using visible spectrophotometer.

44. S.NO pH %Adsorption
1 4 55.8
2 7 56.6
3 9.3 64.1
4 12 58.8

45. By varying the temperature of
solution
• From the above 3 parameter studies it was
observed that dosage of chitosan , agitation time
and pH of solution are optimum at 2 g ,60
minutes and 9.3.
• By varying the temperature of solution ranging
from 26˚C-50˚C.

46. • All the parameters are kept constant such as:
• Time = 60 minutes
• Conc. of CuSO4 = 0.001 N
• Dosage of chitosan = 2 g
• pH = 9.3
• These samples are then kept in shaker for about
60 min and after completion the samples are
analyzed using visible spectrophotometer.

47. S.NO Temp.(˚C) %Adsorption
1 20 64.1
2 25 64.1
3 30 60
4 40 55.8
5 50 51.6

48. • From the above 4 parameter studies it was
observed that dosage of chitosan , agitation
time, pH of solution and temperature of solution
are optimum at 2 g ,60 minutes , 9.3 and 25 ˚C.
• By varying the size of the adsorbent ranging from
16 to 50-.
By varying the size of the
adsorbent

50. • All the parameters are kept constant such as:
• Time = 60 minutes
• Conc. of CuSO4 = 0.001 N
• Dosage of chitosan = 2 g
• pH = 9.3
• Temperature = 25˚C
• These samples are then kept in shaker for about
60 min and after completion the samples are
analyzed using visible spectrophotometer.

51. ˚C
˚C
S.NO Mesh Size %Adsorption
1 50 64.1
2 20 61.6
3 18 57.5
4 16 53.3

52. Adsorption isotherms
• Adsorption isotherm is a curve that expresses the
variation in the amount of adsorbate adsorbed by
the adsorbent with the constant temperature.
• Types of adsorption isotherm are:
• 1) Freundlich Isotherm
• 2) Langmuir Isotherm

53. Freundlich Isotherm
• Freundlich isotherms assumes heterogeneous surface
energies, in which the energy term varies as a function
of surface coverage, in linearized form the freundlich
isotherm is written as:
log qe = log kf + (1/n) × log Ce
where kf and n are freundlich constants

54. logCe logqe
1.66 1.57
2.38 2.27
2.72 2.55
2.94 2.69
3.08 2.81

55. • From the graph, it shows that the adsorption process
follows freundlich isotherm and adsorption process is
physical adsorption.
• The obtained equation is as follows:
y = 0.871x + 0.151
R² = 0.995
• It is a favourable condition for adsorption

56. By solving the above equation the following values
are obtained:
kf =1.416 (mg/g)
n =1.148

57. Langmuir Isotherm:
• Langmuir isotherm assumes monolayer adsorption on to a
surface containing finite number of adsorption sites.
• The linear form of Langmuir isotherm is given as:
Ce/qe = (1/qmax)×b + (Ce/qmax)
Where qe, kf are Langmuir constants related to adsorption
capacity and rate of adsorption respectively.

58. Ce qe/ Ce
45.71 1.23
239.88 1.29
524.81 1.48
870.06 1.75
1206.26 1.94

59. • The above graph the obtained equation is as
follows:
y = 0.000x + 1.166
R² = 0.991
• As the slope is zero qmax = ∞, which is not a
favourable condition. Therefore the adsorption
does not follow Langmuir isotherm.

60. Optimization using response
surface methodology
• Response-surface designs are the only designs
provided that allow for more than two levels.
There are two general types of response-surface
designs.
1) Central Composite design(CCD)
2) Box Behnken design(BBD)

61. . Central Composite design(CCD)
• Regular CCD’s have 5 levels for each factor. This
can be modified by choosing an axial distance of
1.0 creating a Face-Centered, Central Composite
design which has only 3 levels per factor.
• The center points are replicated to provide
excellent prediction capability near the center
of the factor space. A central composite design is
a common augment from the two-level factorial
design.

62. Box Behnken Design
• In Box Behnken design each factor or independent
variable is placed at one of three equally spaced values,
usually coded as −1, 0, +1. (At least three levels are
needed for the following goal).
• The design should be sufficient to fit a quadratic model,
one containing squared terms, products of two factors,
linear terms and an intercept.
• The ratio of the number of experimental points to the
number of coefficients in the quadratic model should be
reasonable.

63. Contour plot:
• It is a plane section of the three dimensional graph
of the function f(x,y) parallel to f(y,z)-plane.
Contour lines are curved, straight, or a mixture of
both the lines on a map describing the intersection
of a real or hypothetical surface with one or more
horizontal planes. The contour plots are as follows:

64. Variation in % adsorption with
temperature and pH of solution

65. • From temperature vs pH contour plot, it was observed
that at the conditions of decreasing temperature,
increasing pH, % adsorption increases and the dark
green colour shows that, the % adsorption is more in
that region. At the condition of Temperature=25ºC and
pH=9.3 more amount of copper ions were removed from
stock solution.

66. Variation in % adsorption with
agitation time and pH

67. • From agitation time vs pH contour plot, it can be
inferred that with increasing agitation time,
increasing pH, % adsorption increases and the dark
green area shows that, the % adsorption is more in
that region. At the condition of agitation time = 60
min and pH = 9.3 more amount of copper ions are
removed from stock solution.

68. Variation in % adsorption with
dosage of adsorbent and pH

69. • From dosage of adsorbent vs pH contour plot, it was
observed that with an increase in dosage of adsorbent
along with the increase in pH, % adsorption increases
and the dark green area reveals that, the % adsorption
is more in that region. At the region of dosage of
adsorbent = 2g and pH = 9.3 more amount of copper
ions are removed from stock solution.

70. Variation in % adsorption with
temperature and agitation time

71. • From temperature of solution vs agitation time
contour plot, it was observed that decreasing
temperature with increasing agitation time increases
the % adsorption and the dark green region indicates
more adsorption when compared to the other
operating conditions. At a temperature of 25ºC and
agitating for 60 min, more amounts of copper ions can
be removed from stock solution.

72. Variation in % adsorption with
temperature and dosage of
adsorbent

73. • From temperature of solution vs adsorbent dosage
contour plot, it was observed that with an decrease
in temperature followed by increase in dosage of
adsorbent, % adsorption increases and the dark green
zone shows that, the % adsorption is more in that
region. At the condition of temperature of solution =
25ºC and dosage of adsorbent = 2g more amount of
copper ions are removed from stock solution.

74. Variation in % adsorption with
agitation time and dosage of
adsorbent

75. • From agitation time vs dosage of adsorbent contour
plot, it was revealed that an increase in agitation
time along with an increase in dosage of adsorbent
results in an increase of % adsorption and the dark
green area clearly indicates more % adsorption. At
the conditions of agitation time = 60min and dosage
of adsorbent = 2g more number of copper ions can be
removed from stock solution.

76. Comparison of optimum values by
experiment and RSM
Parameters Experimental RSM
Agitation time 60 min 64.444 min
Dosage of chitosan 2 g 2.5303 g
pH 9.3 10
Temperature 25ºC 19ºC

77. Conclusion:
• In this study, using the chitosan extracted from prawn
shell waste, adsorption of copper ions was performed.
• For 200g of prawn shell waste 27.75% (55.5g) of
chitosan was extracted.
• A test for chitosan by UV-Visible spectrophotometer
has shown favourable results by confirming the
extraction (existence) of chitosan.
• The extracted chitosan was further used as a potential
adsorbent besides its wide range of applications
(flocculating/coagulating agent) in the treatment of
waste water.

78. • From the experimentation, it was observed that, an
increase in dosage of adsorbent(fine sized) till 2g by
agitating the stock solution for 60min maintaining a
pH of nearly 9.3 at room temperature of 25ºC results
in the %adsorption of 64.1%.
• Throughout the experimentation a constant
concentration of 0.001N of copper solution is
maintained (as per the permissible limits by USEPA).

79. • The adsorption phenomenon is observed to be
physisorption and freundlich isotherm best fits the
experimental data obtained.
• The obtained optimum values from the experimentation
were well matched with the data obtained from RSM
with a correlation coefficient of 0.995

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82. Thank you