MATERIALS AND METHODS

4. MATERIALS AND METHODS

4.1 MATERIALS

Table 4: List of materials

Materials suppliers
Glimepiride HETRO labs,Hyderabad,India.
Polyethylene glycol 4000 S.D. Fine Chem. Ltd, Mumbai, India
Urea S.D. Fine Chem. Ltd, Mumbai, India
MCC S.D. Fine Chem. Ltd, Mumbai, India
Xanthum gum S.D. Fine Chem. Ltd, Mumbai, India
Guar gum S.D. Fine Chem. Ltd, Mumbai, India
Magnesium stearate S.D. Fine Chem. Ltd, Mumbai, India
Talc S.D. Fine Chem. Ltd, Mumbai, India
4.2 EQUIPMENTS
Table 5 List of equipments
Equipments Model /Company
Tablet compression machine Cadmach
Hardness tester Monsanto hardness tester
Dissolution test apparatus Lab India
Disintegration test apparatus Campbell Electronics
Friability test apparatus Riche Rich
U.V visible spectrophotometer Shimadzu UV-1601, Japan
Hot air oven Lab india
pH meter Citizen, India

4.3.1 Development of calibration curve for GLIMEPIRIDE

Methodology
Preparation of Standard Curve for Glimepiride
Preparation of 6.8pH phosphate buffer:
Add 0.2% of SLS for above buffer. And used for further dilutions and makeup.

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 MATERIALS AND METHODS

Determination of Standard Curve

a) Stock solution of 1000μg/ml of Glimepiride was prepared by dissolving
25mg of drug in small quantity of methanol and diluted with methanol to
25ml.
b) From this take 10ml and make up to 100ml using b u f f e r to get a stock
solution of 100 μg/ml.
c) From the above solution take 0.4, 0.8, 1.2, 1.6, 2.0ml and dilute
to 10 ml with buffer to get a concentrations of 4μg/ml, 8μg/ml, 12μg/ml,
16 μg/ml, 20 μg/ml.
d) The absorbance of the different diluted solutions was measured in a UV
spectrophotometer at 232nm.
A calibration curve was plotted by taking concentration of the solution in µg/ml on X-axis
and absorbance on Y-axis and correlation co-efficient “r2” was calculated.

4.4 SOLID DISPERSION OF GLIMEPIRIDE WITH UREA AND

POLYETHYLENE GLYCOL 4000 (PEG 4000)
4.4.1 Methods of Preparation of Solid Dispersion
Solid dispersions were prepared fusion method.
Fusion Method: Each of water soluble carrier UREA, PEG 4000 were weighed
accurately in various ratios (1:1, 1:2, 1:3, 1:4) and melted in a porcelain dish at 80-85ºC
and to this calculated amount of glimepiride was added with thorough mixing for 1-2
minutes followed by quick cooling. The dried mass was then pulverized by passing
through sieve no.85 and stored in a dessicator until used for further studies.
Table 6 Composition of GLIMEPIRIDE solid dispersions
Solid dispersion Method Drug-Polymer Formulation Code
composition ratio

Glimepiride: Urea Fusion method 1:1 SDUF1

1:2 SDUF2

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 MATERIALS AND METHODS

1:3 SDUF3
1:4 SDUF4
Fusion method 1:1 SDPF1
Glimepiride: PEG 1:2 SDPF2
4000 1:3 SDPF3
1:4 SDPF4

4.5 Characterization of GLIMEPIRIDE Solid Dispersions

4.5.1 Fourier Transform Infrared spectroscopy (FT-IR)
The samples of U r e a , PEG 4000 and solid dispersion were prepared in the form
of KBr pellets and subjected for scanning from 4000 cm-1 to 400 cm-1 using
FT-IR spectrophotometer.
4.5.2 Drug content
An accurately weighed quantity of solid dispersion equivalent to 32mg of
GLIMEPIRIDE was taken into a 100ml volumetric flask, dissolved in acetone and suitably
diluted with 6.5 pH phosphate buffer. The content of GLIMEPIRIDE was determined
spectrophotometrically at 232 nm against suitable blank using UV-visible spectrophotometer
(1601, Shimadzu, Kyoto, Japan).

4.5.3 IN vitro dissolution studies

The quantity of solid dispersion equivalent to 32mg of GLIMEPIRIDE was filled in
colourless hard gelatin capsule by hand filling method. The dissolution study of capsules
was conducted using dissolution testing USP apparatus 1 (basket method) in 900 ml of 6.8
pH phosphate buffer and to this buffer add 0.2% SLS at 37±0.5ºC and at a speed of
50 rpm. Aliquot of 5ml was withdrawn at predetermined time interval and equivalent
amount of fresh medium was replaced to maintain a constant volume after each
sampling and analyzed spectrophotometrically at 232nm against suitable blank using
UV-visible spectrophotometer.

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