CHAPTER 6 RESULTS AND DISCUSSION

6.0 RESULTS AND DISCUSSION

a) Organoleptic properties:
Timolol was studied for organoleptic properties such as color, odor.

Color: White

Odor: Odorless

b) Melting point

Table No 10 - Melting point of drug and polyelectrolyte by capillary method

Sr. Sample M.P. Standard M.P. Observed

No.
1. Timolol 201.5-202.5 oC 201 oC
2. Gellan Gum 85-90 oC 87 oC

Melting point of Timolol and Gellan gum was determined by using capillary
method. The melting point of Timolol and Gellan Gum was found 201 oC and
87oC respectively that is within a standard range from this we can conclude that
drug and polyelectrolyte both were in pure form.

c) Special Characterizaation
I. The λmax determination

λ max of Timolol was found to be 294nm in STF pH 7.4.

II. FT-IR spectra of Timolol Maleate

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CHAPTER 6 RESULTS AND DISCUSSION

The FT-IR Spectrum of Timolol Maleate pure drug was found to be similar to the
standard spectrum of Timolol Maleate showed the following functional groups at their
frequencies.

Table No. 15 - Structure elucidation of Timolol Maleate

Sr. No. Functional Groups Standard Peaks Observed Peaks

(cm-1) (cm-1)
1. COH stretch (s) 1072

2. COC stretch (s) 1120

3. NH stretch (m,w) 3298

4. OH stretch (s) 3545

Figure No : FTIR spectra of Timolol

Table No : Structure elucidation of Gellan Gum

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CHAPTER 6 RESULTS AND DISCUSSION

Figure No : FTIR Spectra of Gellan Gum

III. Experimental Methods

a. Calibration curve in water

Table No 11 - Calibration curve of Timolol in distilled water

Sr No. Conc. (PPM) Absorbance at 295 nm

1. 2 0.053
2. 4 0.104
3. 6 0.156
4. 8 0.208
5. 10 0.258

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CHAPTER 6 RESULTS AND DISCUSSION

Figure No : Calibration curve of Timolol in distilled water

The calibration curve of Timolol was taken in distilled water. The various concentrations
were prepared and absorbance was measured at 294 nm and absorbance vs concentration
graph was plotted as given in figure no .

b. Calibration curve of Timolol in Stimulated Tear Fluid

Table No. 12 – Calibration curve of Timolol in Stimulated Tear Fluid (pH7.4 )

Sr No. Conc. (PPM) Absorbance at 294 nm

1. 0 0
2. 5 0.129
3. 10 0.230
4. 15 0.373
5. 20 0.458
6. 25 0.571
7. 30 0.717
8. 35 0.815
9. 40 0.964

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y = 0.023x + 0.001
0.8 R² = 0.999
0.7
0.6
 0.4
0.3
CHAPTER0.2
6 RESULTS AND DISCUSSION
0.1
0
Figure No 6: Calibration
0 10
curve20of Timolol
30
in Stimulted
40
Tear Fluid (pH7.4 )
50

Concentration ( μg/ml)
The calibration curve of Timolol was taken in artificial Tear Fluid . The various
concentrations were prepared and absorbance was measured at 294 nm and absorbance vs
concentration graph was plotted as given in figure no 6.

d) Solubility study of Timolol

Solubility of Timolol was determined in different solvents i.e. distilled water, ethanol,
Methanol,Artificial tear fluid at pH 7.4 at 37˚C by using orbital shaking incubator with 50
rpm. The solubility of Timolol in different solvents showed in table no .

Table No : Solubility of Timolol in different solvents

Sr No. Solvent Solubility

(Mg/ml)

1. Distilled water 0.269

2. Ethanol Soluble
3. Methanol Soluble
4. Chloroform Sparingly Soluble
5 STF (PpH 7.4) Soluble

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CHAPTER 6 RESULTS AND DISCUSSION

6.2 Evaluation of In-Situ Gel

a) Visual appearance :
During preparation of in situ gel of timolol drug , visual appearance of formulation on
varing conc of polymer in drug has been observed. The visual Appearnce of timolol

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CHAPTER 6 RESULTS AND DISCUSSION

ocular gel is displayed in table . The visual appearance of various formulations was
Transparent.
Table No : Visual Apperance Analysis of Formulations

Formulations Visual Appearance

F1 Transparent
F2 Transparent
F3 Transparent
F4 Transparent
F5 Transparent

b) Clarity

Formulations Clatity
F1 Clear
F2 Clear
F3 Clear
F4 Clear
F5 Clear
F6 Clear

c ) pH

The pH of various formulations observed and display in Table .The pH of formulations

are found to be in range of 6.5 to 6.7 which is good for eye .All formulations of ocular
gel were shown pH near to eye.

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CHAPTER 6 RESULTS AND DISCUSSION

Table No: pH Analysis of Formultions

Formulations pH
F1 7.12
F2 7.24
F3 7.26
F4 7.25
F5 7.31
F6 z

d) Drug Content
Formulation Drug Content
F1 98.01
F2 97.06
F3 96.08
F4 95.29
F5 93.43
F6 92.97

e)In vitro Gelation

The Gelling Capacity was measured for formulations F1 to F5 Formulations and display in
table . The gelling capacity of the prepared formulations was determined by placing a drop
of formulations in vial containing 2 ml freshly prepared simulated tear fluid and visually
observed.
Table No: Gelling capacity
Formulations Gelling Capacity
F1 ++
F2 +++
F3 +++
F4 +++
F5 +++

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CHAPTER 6 RESULTS AND DISCUSSION

Note - ++ gelation immediate and remain for few hours, +++ shows gelation immediate
and remain for extended periods.
In Vitro Drug Released :

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