4.

RESULTS AND DISCUSSION

4.1. Preformulation Studies

4.1.1. Dissolution Method Development

The results of dissolution studies of 25 mg of standard Carvedilol and the marketed

products with water, and SLS in water, are given in Tables 3 to 5. The results indicate
that less than 3% of Carvedilol (standard) dissolves in water in 60 minutes, whereas in
the case of marketed it Carvedilol was around 30 %. A significantly higher dissolution
was obtained, however, in presence of SLS. An increase in the rate and extent of
dissolution was seen with an increase in SLS concentration. The increase was only
marginal when the SLS concentration was increased from 2.0 to 3.0 %. The results
indicate that dissolution was optimal (i.e. > 85 %) when the concentration of SLS was 1
% and rpm was 50. This was therefore, selected for further studies.

Table .Dissolution tests for standard Carvedilol

(Single time point)

S.No. Medium Parameters Time %

dissolved
1 Water 100rpm, Paddle 60 2.15 %
2 Sodium Lauryl Sulphate 3 % 100rpm, Paddle 60 94.53%
in water
3 Sodium Lauryl Sulphate 2% 100rpm, Paddle 60 93.82%
in water
4 Sodium Lauryl Sulphate 1% 100rpm, Paddle 60 88.99%
in water
5 Sodium Lauryl Sulphate 0.5% 100rpm, Paddle 60 78.04 %
in water
6 Sodium Lauryl Sulphate 100 rpm, Paddle 60 78.30%
0.25% in water
 Table . Dissolution tests for standard Carvedilol
(Different time points)

S.No. Medium Parameters Time %

(minutes) dissolved
1 Sodium Lauryl Sulphate 1% 100rpm, Paddle 30 75.99
in water 45 86.47
60 90.93
2 Sodium Lauryl Sulphate 1% 50 rpm, Paddle 30 79.41
in water 45 86.58
60 88.74
3 Sodium Lauryl Sulphate 0.5 50 rpm, Paddle 30 73.03
% in water 45 80.75
60 82.38
4 Sodium Lauryl Sulphate 50 rpm, Paddle 30 61.43
0.25% in water 45 71.51
60 78.52

Table . Dissolution tests for marketed Carvedilol tablets

(Cardivas, 25 mg, Sun pharma)

S.No. Medium Parameters Time % dissolved

(minutes)
1 Water 50 rpm, Paddle 60 25.27
2 Sodium Lauryl Sulphate 50 rpm, Paddle 10 71.38
0.5% in water 20 93.38
30 98.32
45 98.55
60 99.18

4.1.2. Development of calibration curve

The λ max for the highest concentration observed was 242.8 nm. At this λ max
absorbance of different concentrations prepared were recorded. The absorbances of
different dilutions are given in Table 6 and the plot of concentration vs. absorbance is
given in Figure 1. The regression coefficient value (r2) obtained was 0.9992 which
indicates a fair linearity.

Table . Calibration curve values for Carvedilol

 Concentration Absorbance
3 0.358
6 0.663
9 1.008
12 1.318
15 1.614

Figure . Calibration curve for Carvedilol

y = 0.1041x + 0.02
CC for Carvedilol
R2 = 0.9992
1.8
1.6
1.4
Absorbance

1.2
1
0.8
0.6
0.4
0.2
0
0 5 10 15 20
Concentration in (mcg/ml)

4.1.3. Compatibility studies

The initial IR spectra of the drug and the polymer are satisfactory with their characteristic
absorption bands. Similarly, the physical mixtures also indicate the presence of
characteristic peaks of the drug and the polymer. It is clear that the drug and the
excipients are free from any significant chemical interactions.

4.2. Development of SR Carvedilol tablets

The granulation properties examined included the angle of repose, loose bulk density,
tapped bulk density, Carr’s index (Compressibility index) and drug content. The
formulated tablets have acceptable hardness and friability, capable of withstanding the
abrasional forces during transit.
Evaluation of Granules
 Tapped
Angle of Loose Moisture
S.No Batch No Bulk Carr’s Index (%)
Repose Density content
Density
1. CDA-1 39o08’ 0.3908 0.4112 04.96 1.23
2. CDA-2 38o38’ 0.4526 0.5246 13.72 1.52
3. CDA-3 35o54’ 0.4635 0.4968 06.70 1.96
4. CDA-4 38o28’ 0.4401 0.5123 14.09 1.96
5. CDA-5 40o42’ 0.4587 0.4869 05.79 2.05
6. CDS-1 27o38’ 0.4452 0.4693 05.14 1.56
7. CDS-2 29o56’ 0.4528 0.5123 11.61 1.54
8. CDS-3 35o42’ 0.4106 0.5968 31.20 1.75
9. CDS-4 32o34’ 0.4176 0.4694 11.04 1.62
10. CDS-5 30o42’ 0.4583 0.5286 13.30 1.96
11. CDL-1 31o22’ 0.4521 0.4639 02.54 1.45
12. CDL-2 30o18’ 0.3944 0.4639 14.98 1.44
13. CDL-3 27o22’ 0.3965 0.4526 12.40 1.86
14. CDL-4 28o28’ 0.4350 0.4996 12.93 1.46
15. CDL-5 29o38’ 0.3494 0.4425 21.04 1.43

A hydrophilic matrix controlled release system is a dynamic system composed of

polymer wetting, hydration and dissolution. In all the batches, it was observed that as the
polymer concentration increase the drug release rate decreases. From T50 values it is
clear that the batches with higher polymer to diluent ratios have higher T 50 values. A
convincing explanation for the decrease in the rate of drug release with increase in the
polymer concentration is due to the increase in the thickness of the gel layer, thus
retarding drug diffusion out of the tablet. At the same time, other soluble excipients will
also wet, dissolve and diffuse while the insoluble ingredients will be held in place until
the polymer erodes or dissolves.
The results of the effect of swelling, insoluble fillers like Avicel PH102 (Microcrystalline
cellulose), starch 1500 and soluble filler (Lactose). The results indicate that these fillers
change the release profiles due to a change in the rate of swelling at the tablet surface.
Faster release rates were obtained with lactose followed by starch 15000 and Avicel Ph
102. This might be due to the controlled swelling of Avicel Ph 102.

The effects of sodium starch glycolate (SSG) on the release of carvedilol indicate that
the rate of release decreases with the decrease in SSG. This might be due to the
reduction in the capillary channels with decrease in the concentration of SSG.

Tablet Characterization
 Average
S.No Batch No Hardness Thickness Weight
(mm)
1. CDA-1 5.3±0.67 3.95±0.04 101.10±1.29
2. CDA-2 5.4±0.55 3.96 ±0.01 101.65±2.02
3. CDA-3 4.8±0.45 3.96±0.02 101.81±2.50
4. CDA-4 5.6±0.55 3.96±0.01 102.60±1.96
5. CDA-5 5.2±0.45 3.96±0.02 102.59±1.86
6. CDS-1 5.4±0.55 3.95±0.03 100.13±1.47
7. CDS-2 5.3±0.84 3.99±0.02 101.43±2.12
8. CDS-3 5.5±0.79 3.99±0.02 103.53±1.69
9. CDS-4 5.3±0.84 3.99±0.03 101.78±2.74
10. CDS-5 5.0±0.61 4.00±0.03 103.28±1.38
11. CDL-1 5.3±0.45 3.92±0.03 102.02±1.14
12. CDL-2 5.2±0.57 3.90±0.03 102.09±0.99
13. CDL-3 5.4±0.65 3.90±0.03 102.48±2.20
14. CDL-4 5.1±0.55 3.88±0.02 103.23±1.51
15. CDL-5 5.2±0.57 3.88±0.02 101.79±1.72

The optimized formulation was CDA-3 containing 15 %w/w of polymer, 1% w/w of

diffusion controller and Avicel PH 101 as filler, subjected to stability studies, toxicity
studies and bioavailability studies in healthy human volunteers.
Figure . Invirto dissolution study of Carvidilol tablets (CDA-1 to CDA-5) in Distilled water
with SLS
125

100

75
% Release

50

25

CDA-1 CDA-2 CDA-3 CDA-4 CDA-5

0
0 2 4 6 8 10 12 14 16 18 20 22 24
Time

Figure . Invirto dissolution study of Carvidilol tablets (CDS-1 to CDS-5) in Distilled water
with SLS

100

75
% Release

50

25

CDS-1 CDS-2 CDS-3 CDS-4 CDS-5

0
0 2 4 6 8 10 12 14 16 18 20 22 24
Time
Figure . Invirto dissolution study of Carvidilol tablets (CDL-1 to CDL-5) in Distilled water
with SLS

125

100

75
% Release

50

25

CDL-1 CDL-2 CDL-3 CDL-4 CDL-5

0
0 2 4 6 8 10 12 14 16 18 20 22 24
Time

4.3. Comparison of In vitro profiles

The profiles of cumulative % release against vs. time for the conventional marketed IR
tablets are as given in Figure . The results reveal that dissolution data is highly
reproducible (as evident from the SD values) and hence only the average values were
plotted.
 Figure . In vitro dissolution profile of Marketed IR tablets

120

100

% release
80

60

40

20

0
0 20 40 60 80
time(minutes)

The in vitro profile of cumulative drug release Vs time for the formulated SR tablet of
Carvedilol is shown in Figure 3. The results reveal that the T50 value of the formulated SR
tablets of Carvedilol is 8.0 h as compared to less than 10 minutes of the marketed IR
tablets. It is, therefore, capable of sustaining the drug release in in vivo. To confirm the
same, the developed formulation was subjected to pilot bioavailability studies in healthy
human volunteers (n=6).

Figure . In vitro dissolution profile of the optimized formulation SR

100
cumulative release %

80

60

40

20

0
0 4 8 12 16 20 24
time(hrs)
4.4. Stability Studies

No significant change was observed for the formulated sustained release matrix tablets
of Carvedilol with respect to its physicochemical parameters and in vitro drug release as
evident by table 7. The developed formulation for Carvedilol is, therefore, stable at
various temperature and humidity conditions for a period of 3 months.

Table . Stability data at the end of three months

Parameter Initial Real time Accelerated

Thickness mm 3.95±0.022 4.00±0.02 3.96±0.04
Hardness (kg/cm3) 4.50±0.316 4.50±0.50 5.00±0.36
Friability (%)(n=1) 0.34 0.21 0.30
Drug content (%) 97.59±0.547 98.50±1.98 97.64±1.80
T50 in vitro (h) 8.0 ± 0.5 8.12 ± 0.45 8.30±0.56

4.5. Toxicity studies

The SR Carvedilol formulation developed was tested for acute oral toxicity in female
Wistar rats as per the OECD guideline for the testing of chemicals, “Acute Oral Toxicity
Study (UP and DOWN Method)”, Guideline No. 425.

Based on the results of the acute oral toxicity of the SR Carvedilol formulation developed
in Wistar rats, it can be concluded that the LD 50 of the test item is greater than
2000mg/kg.

The results of the study repeated dose (28 days) oral toxicity study indicate that the
developed SR Carvedilol formulation has no adverse effect on general health, growth,
hematological and clinical chemistry parameters and organ weights of Wistar rats at a
dose of 18 mg/kg b.wt. No dose related changes were observed in any of the
parameters evaluated. From these findings, the evaluated No Observed Adverse Effect
Level (NOAEL) of Carvedilol SR formulation in Wistar rats is 18 mg/kg b.wt. under the
testing conditions and doses employed.

From the in vivo micronucleus test in mice it can be concluded that the developed
Carvedilol SR formulation do not produce genotoxicity at the dose employed and under
the testing conditions.
 4.6. Bioavailability studies
The results of the pharmacokinetic analysis reveal that the developed Carvedilol SR
Tablets containing 25 mg of Carvedilol is capable of sustaining the drug release in in
vivo and can overcome the draw backs associated with conventional therapy of
Carvedilol. It has, however, to be confirmed by conducting the bioavailability studies in
larger population.

3.2. Evaluation of marketed IR tablets

The product details of the marketed immediate release (IR) tablet formulation of
Carvedilol used for in vitro drug release evaluation is given in Table 1.

Table . Product details of marketed conventional formulation

Trade Name Cardivas

Strength 25.00mg
Batch. No AD 40263
Mfg Oct ‘2004
Exp Sep’2006
Price Rs.82.85/10tablets(L.T. extra)
Packing Strips of 10
Manufactured by Sun Pharmaceutical Industries
Dadra – 396 191, India

Dissolution conditions
Apparatus : USP Type II; Paddle
RPM : 50
Medium : 900 ml of 1 % SLS in water
Sampling volume : 5 ml
Sampling intervals : 10, 20, 30, 45 and 60 min
Analysis : UV; λ max, 242.8 nm

Formulation parameters optimized were,

• Concentration of the polymer,
• Concentration and type of filler and
• Concentration of release modifier.
Formula for Carvediliol Tablets
Table . Composition of Carvedilol SR tablets