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International Journal for Pharmaceutical
Research Scholars (IJPRS) V-3, I-2, 2014 ISSN No: 2277 - 7873
RESEARCH ARTICLE
© Copyright reserved by IJPRS Impact Factor = 1.0285 17
Development and Validation of Analytical Method for Simultaneous Estimation of
Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form Patel PD, Makwana CJ, Patel MD, Dey S, Upadhyay UM
Sigma Institute of Pharmacy Vadodara, Gujarat, India.
Manuscript No: IJPRS/V3/I2/00158, Received On: 01/04/2014, Accepted On: 05/04/2014
ABSTRACT
A simple, economic, selective, precise, and stability-indicating Reverse phase High Performance Liquid
Chromatography method for analysis of Vardenafil and Dapoxetine HCl was developed and validated
according to ICH guidelines. The quantification of the drug was carried out using Hypercil BDS C18
250mm × 4.6mm × 5μm or its equivalent in isocratic mode, with mobile phase compressing of Buffer
(KH2PO4): Acetonitrile: Triethylamine (55: 45: 0.1 v/v/v) the flow rate was 1ml/min and the detection
was carried at 266 nm. The retention time for Vardenafil and Dapoxetine HCl was found to be 3.673 and
5.070min respectively. The percent assay for Vardenafil and Dapoxetine HCl was found to be 100.22%
and 99.44% respectively. The method was also applied for the determination of Vardenafil and
Dapoxetine HCl in the presence of their degradation products formed under variety of stress conditions.
Proposed method was validated for precision, accuracy, linearity range, specificity and robustness.
KEYWORDS
Vardenafil, Dapoxetine HCl, RP-HPLC, Stability Indicating, Validation
INTRODUCTION
Dapoxetine Hydrochloride
Category: Selective serotonin reuptake inhibitor.
Empirical Formula C21H23NO ·HCl, IUPAC
Name (S)-N,N-dimethyl-3-(naphthalen-1-
yloxy)-1-phenylpropan-1-amin.
Mechanism of Action Dapoxetine hydrochloride
is a SSRI in development for the management of
premature ejaculation. Its mechanism of action
is to increase the serotonin level in central
nervous system.
Vardenafil
Category: Vasoconstrictor Agents,
Phosphodiesterase Inhibitors, Anti-Impotence
Agents
*Address for Correspondence:
Paritosh D. Patel
Sigma Institute of Pharmacy, Bakrol, Ajwa-Nimeta Road,
Vadodara- 390019, Gujarat, India .
E-Mail Id: paritosh4u4ever@gmail.com
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 18
Empirical Formula C23H32N6O4S, IUPAC Name
2-[2-ethoxy-5-(4-ethylpiperazine-1-
sulfonyl)phenyl] -5-methyl- 7-propyl-1H,
4Himidazo[4,3-f][1,2,4]triazin-4-one.
Mechanism of Action Vardenafil inhibits the
cGMP specificphosphodiesterase type 5 (PDE5)
which is responsible for degradation of cGMP
in the corpus cavernosum located around the
penis. Penile erection during sexual stimulation
is caused by increased penile blood flow
resulting from the relaxation of penile arteries
and corpus cavernosal smooth muscle. This
response is mediated by the release of nitric
oxide (NO) from nerve terminals and
endothelial cells, which stimulates the synthesis
of cGMP in smooth muscle cells. Cyclic GMP
causes smooth muscle relaxation and increased
blood flow into the corpus cavernosum. The
inhibition of phosphodiesterase type 5 (PDE5)
by vardenafil enhances erectile function by
increasing the amount of Cgmp.
MATERIALS AND METHOD
Materials
Gift sample of Vardenafil was procured from
Centurion Laboratory Pvt. Ltd. Vadodara.
Dapoxetine HCl was kindly gifted by Centurion
Laboratory Pvt. Ltd., Vadodara both the drugs
are used as a standard without further
purification. HPLC grade Methanol,
Acetonitrile, KH2PO4, Ortho-phosphoric acid
and Double distilled water were used.
Method
Identification of Drugs
Identification of both the drug should be done
by following parameter:
Melting point
IR identification
Solubility
Selection of Wavelength
The sensitivity of HPLC method that uses UV
detection depends upon proper selection of
detection wavelength. An ideal wavelength is
the one that gives good response for the drugs
that are to be detected. In the present study 10
μg/ml of VAR and 30 μg/ml of DAPO. These
drug solutions were than scanned in the UV
region of 200-400 nm and the overlay spectrum
were recorded.
Selection of Chromatographic Condition
Proper selection of the HPLC method depends
upon the nature of the sample (ionic, ionizable
or neutral molecule), its molecular weight and
solubility. The drugs selected for the present
study are polar in nature and hence either
reversed phase or ion-pair or ion exchange
chromatography can be used. Reversed phase
HPLC was selected for the initial separations
because of its simplicity and suitability. To
optimize the chromatographic conditions, the
effect of chromatographic variables such as
mobile phase, pH, flow rate, and solvent ratio
were studied. The resulting chromatograms
were recorded and the chromatographic
parameters such as capacity factor, asymmetric
factor, and resolution and column efficiency
were calculated. The conditions that gave the
best resolution, symmetry and capacity factor
were selected for estimation.
Effect of Ratio of Mobile Phase
Mixed standard solution containing 10 ppm
Vardenafil and 30 ppm Dapoxetine HCl was
chromatographed using different type of mobile
phases and optimized mobile phase was
finalised.
Preparation of Mobile Phase
A mixture of 45 ml Acetonitrile and 55 ml
0.02m Phosphate Buffer (pH 3.5+0.1% Tea) of
HPLC grade filtered through 0.75 μm filter
paper, Sonicate for 15minutes to degas the
mixture and used as mobile phase.
Preparation of Standard Stock Solution
Vardenafil Standard Stock Solution
(100μg/ml)
A 10 mg of standard Vardenafil was weighed
and transferred to a 100 ml volumetric flask and
dissolved in 25 ml mobile phase. The flask was
shaken and volume was made up to the mark
with mobile phase to give a solution containing
100μg/ml Vardenafil. Dapoxetine HCl standard
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 19
stock solution: (300μg/ml) A 30 mg of standard
Dapoxetine HCl was accurately weighed and
transferred to a 100 ml volumetric flask and
dissolved in 25 ml mobile phase. The flask was
shaken and volume was made up to the mark
with mobile phase to give a solution containing
300μg/ml Dapoxetine HCl.
Calibration Curve for Vardenafil and
Dapoxetine HCl
Calibration curve for the Vardenafil (5-15
μg/ml) and Dapoxetine HCl (15-45 μg/ml)
Appropriate volume of aliquots from standard
Vardenafi and Dapoxetine HCl stock solutions
weretransferred to same volumetric flasks of 10
ml capacity. The volume was adjusted to the
mark with mobile phase give a solution
containing 5, 7.5, 10, 12.5 and 15 μg/ml
Vardenafil and 15, 22.5, 30, 30.7, and 45 μg/ml
Dapoxetine Hcl.
The mixed standard solution was
chromatographed for 10minutes using mobile
phase at a flow rate of 1.0 ml/min. The graphs
were plotted for peak area vs. concentration for
both thedrugs.
Determination of Vardenafil and Dapoxetine
HCl from its Marketed Formulation
Ten tablets were weighed and average weight
was calculated. The tablets were crushed to
obtain fine powder. Tablet powder Weighted
equivalent to 25mg was transferred to 100 ml
volumetric flask. Mobile Phase was added to
dissolve the drugs and sonicated for 10 minutes
and then volume was made up to the mark with
same.
The solution was then filtered through
Whatmann filter paper (No. 41). From that 1 ml
of aliquots was taken and diluted up to 10 ml for
getting concentration of 100 μg/ml. From this
stock solution, working standard solution of 10
μg/ml of VAR and 30 μg/ml of DAPO was
prepared and diluted it up to 10 ml with mobile
phase. This solution was used for the estimation
of VAR and DAPO in their combined dosage
form.
Sample Preparation
Estimation of Vardenafil and Dapoxetine HCl in
its Marketed formulation: The prepared sample
solution was chromatographed for 10 minutes
using mobile phase at a flow rate of 1.0 ml/min.
From the peak area obtained in the
chromatogram, the amounts of both the drugs
were calculated by fitting peak area responses
into the equation of the straight line representing
the calibration curves for Vardenafil and
Dapoxetine HCl.
Validation of the Development HPLC
Method
Accuracy
Preparation of sample solution for % recovery
10 mg Vardenafil and 30 mg Dapoxetine HCl
was accurately weighed and transferred to
volumetric flask of 100ml capacity and aliquot
them to make final concentration10μg/ml
Vardenafil and 30μg/ml Dapoxetine HCl. The
resulting solution was filtered through Whatman
filter paper. Then chromatogram of each sample
solutions was taken at selected wavelength for
Vardenafil and Dapoxetine HCl and
concentration is calculated which is known as
pre-analyzed sample.
Table 1: Spiking of Std. solution in pre-analyzed
solution
Spiking
Concent
ration
Concentration
Of Preanalysed
Sample (μg/ml)
Concentratio
n of Spiked
Std. Solution
(μg/ml)
Total
Concentrati
on (μg/ml)
VAR DAPO VAR DAPO VAR DAPO
80 10 30 8 24 18 54
100 10 30 10 30 20 60
120 10 30 12 36 22 66
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 20
Precision
Repeatability
6 replicates of standard mixture solution having
Vardenafil (10 μg/ml) and Dapoxetine HCl
(30μg/ml) were prepared and chromatograms
were recorded and RSD was calculated.
Intraday Precision
Standard solutions containing 5, 10 and 15
μg/ml Vardenafil and 15, 30 and 45 μg/ml
Dapoxetine HCl were analyzed 3 times on the
same day as per the procedure. Chromatogram
of each sample was taken. SD and RSD were
calculated.
Inter day precision
Standard solutions containing 5, 10 and 15
μg/ml Vardenafil and 10, 30 and 60 μg/ml
Dapoxetine HCl were analyzed on three
different daysas per the procedure.
Chromatogram of each sample was taken. SD
and RSD were calculated.
Linearity and Range
The linearity peak area response was determined
by analyzing solutions having concentrations in
the range of 5-15μg/ml and 15-45 μg/ml for
Vardenafil and Dapoxetine HCl respectively
from same solution.
Limit of Detection
Calibration Curve was repeated for 6 times and
the SD of the Intercept was calculated then LOD
was calculated.
Limit of Quantitation
Calibration Curve was repeated for 6 times and
the SD of the Intercept was calculated then LOD
was calculated.
Robustness
Robustness study was performed in following
altered chromatographic conditions:
• Variation in Analyst
• Variations in the mobile phase (± 1)
• Variation in pH (± 0.1)
Forced Degradation Study (Stress Testing)
by RP-HPLC
Preparation of Stock Solution from Marketed
Formulation
Twenty capsules were taken and drug content
was collect by empty gelatin shell. Capsule
powder equivalent to 10 mg VAR and 30 mg of
DAPO was taken in 100 ml volumetric flask.
Mobile phase (50 ml) was added to the above
flask and was sonicated for 15 minutes. The
solution was filtered using 0.45 μm whatman
filter paper and volume was made up to the
mark with the mobile phase. It was
concentration of 100 μg/ml of VAR and 300
μg/ml of DAPO.
Forced degradation study was carried out by
Acid Hydrolysis, Alkali Hydrolysis, Oxidative
Hydrolysis, Thermal Hydrolysis and Photolytic
Hydrolysis.
RESULTS AND DISCUSSION
Identification of Drugs
Melting Point
Table 2: Melting Point of API
Name of
Drug
Observed
M.P (0C)
Reported
M.P (0C)
Vardenafil 170-174 (0C) 175-179 (
0C)
Dapoxetine
HCl 228-233 (
0C) 230-235 (
0C)
IR Identification by FTIR Spectra
Figure 1: FTIR spectrum of Vardenafil
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 21
IR spectrum of Vardenafil sample was
interpreted. From these, we can say that the
given sample may be Vardenafil
Figure 2: FTIR spectrum of Dapoxetine HCl
IR spectrum of Dapoxetine HCl sample was
interpreted. From these, we can say that the
given sample may be Dapoxetine HCl.
Solubility Study
Solubility
Vardenafil Dapoxetine HCl
Soluble in Distilled
water, Ethanol (98%),
Methanol, Phosphate
Buffer and
Acetonitrile
Soluble in Methanol,
Acetonitrile, Ethanol,
Phosphate Buffer and
Distilled water
Selection of Wavelength
Vardenafil and Dapoxetine HCl were scanned in
UV as well different trails were taken in RP-
HPLC at different wavelength in which both
Vardenafil and Dapoxetine HCl show
reasonably good response at 266.0nm.
Figure 3: Spectra of Vardenafil and Dapoxetine
HCl at 266 nm
Selection of Mobile Phase
Different type of mobile phase were tried and
from chromatogram optimized mobile phase
was finalised having the composition as below.
Buffer (0.02m KH2PO4 (pH-3.5+0.1% TEA)-
Acetonitrile (55-45 v/v)
Figure 4: Buffer 0.02m KH2PO4 (pH
3.5+0.1%TEA): CAN (55:45) (Finalized Mobile
phase)
Calibration Curve for the Vardenafil and
Dapoxetine HCl
Figure 5: Chromatograms of Vardenafil and
Dapoxetine HCl for Linearity
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 22
Calibration Curve for the Vardenafil (5-15
μg/ml)
Figure 6: Graph of Calibration curve for
Vardenafil
Table 4: Calibration curve for Vardenafil
Concentration
(μg/ml)
Peak Area of
Vardenafil
5 684.863
7.5 1023.604
10 1366.508
12.5 1657.954
15 2046.418
Linearity range for Vardenafil was found to be
5-15 μg/ml in Mobile Phase. Regression
Equation for Vardenafil 266 nm: Y=134.3x +
12.88. r2 value: 0.998.
Calibration Curve for the Dapoxetine HCl
(15-45 μg/ml)
Linearity range for Dapoxetine HCl was found
to be 15-45 μg/ml in mobile phase. Regression
Equation for Dapoxetine HCl at 266 nm: Y=
81.25x + 1.993 r2 value: 0.999.
Figure 7: Graph of Calibration curve for
Dapoxetine HCl
Calibration curve for the Dapoxetine HCl (15-
45 μg/ml)
Table 5: Calibration curve for Dapoxetine HCl
Concentration
(μg/ml)
Peak Area of
Dapoxetine Hcl
15 1221.746
22.5 1826.255
30 2438.193
37.5 3060.792
45 3651.643
% Assay of Vardenafil (10µg/ml) and
Dapoxetine HCl (30µg/ml) in their Marketed
Product
% Assay of Vardenafil and Dapoxetine HCl was
found in an acceptance limit as shown in table 6.
So this method could be used for analysis of this
combination.
Validation of the developed HPLC method
Accuracy (% Recovery Study)
Vardenafil:
Concentration of Preanalysed sample of
Vardenafil: 10μg/ml.
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 23
Result obtained reveals that % recovery of
Vardenafil was within acceptance criteria given
in ICH i.e. 98-102%.
Dapoxetine HCl:
Concentration of Preanalysed sample of
Dapoxetine HCl: 30 μg/ml
Result obtained reveals that % recovery of
Dapoxetine HCl was within acceptance criteria
given in ICH i.e. 98-102%.
Table 6: % Assay of Vardenafil and Dapoxetine HCl
Drug
Concentration
in Marketed
Formulation
Concentration
taken for %
Assay
Peak
Area of
Sample
Solution
Concentration
found from
marketed
Formulation
% Assay ±
SD
Vardenafil 10 mg 10 μg/ml 1463.61 10.80 100.22 ± 0.42
Dapoxetine
Hcl 30 mg 30 μg/ml 2427.08 29.80 99.44 ± 0.32
Table 7: % Recovery of Vardenafil
Level of
Recovery
Amount of
Std. VAR
Spiked (μg/ml)
Total
Amount of
VAR (μg/ml)
Amount of
VAR
recovered (μg/ml)
% Recovery
Mean %
Recovery ±
SD
80 %
8 18 7.98 99.856
99.762 ± 0.70 8 18 8.03 100.415
8 18 7.92 99.015
100 %
10 20 9.97 99.796
99.862 ± 0.84 10 20 9.89 98.998
10 20 10.00 100.684
150 %
12 22 11.90 99.810
99.946 ± 0.75 12 22 12.00 100.312
12 22 11.96 99.717
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 24
Table 8: % Recovery of Dapoxetine HCl
Level of
Recovery
Amount of
Std. DAPO
Spiked (μg/ml)
Total
Amount of
DAPO (μg/ml)
Amount of
DAPO
recovered (μg/ml)
% Recovery Mean %
Recovery ± SD
80 %
24 54 24.03 100.130
99.926 ± 0.64 24 54 24.10 100.450
24 54 23.80 99.199
100 %
30 60 29.99 99.973
100.108 ± 1.0 30 60 29.75 99.175
30 60 30.35 101.177
150 %
36 66 36.19 100.548
100.295 ± 0.37 36 66 36.16 100.471
36 66 35.95 99.866
Precision
Repeatability
Table 9: Repeatability data for Vardenafil and Dapoxetine HCl
Standard
Drug
Target
Concentration
(μg/ml)
Peak
Area of
Sample
Found
Concentration
(μg/ml)
Mean SD % RSD
Vardenafil
10 1352.853 9.9
10 0.12 1.2
10 1381.578 10.1
10 1352.450 9.9
10 1385.690 10.2
10 1351.490 9.9
10 1358.773 10.0
Dapoxetine
Hcl
30 2465.094 30.3
29.9 0.22 0.73
30 2413.817 29.7
30 2423.541 29.8
30 2472.380 30.0
30 2411.389 29.7
30 2433.313 29.9
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 25
The SD and % RSD for Repeatability of both
the drugs was found to be less than 1 and less
than 2 respectively.
So, it was concluded that proposed method for
estimation of Vardenafil and Dapoxetine HCl is
précised in nature.
Intraday Precision
The SD and % RSD for Intraday precision of
both the drugs was found to be less than 1 and
less than 2 respectively. So, it was concluded
that proposed method for estimation of
Vardenafil and Dapoxetine HCl is précised in
nature.
Table 10: Intraday precision data for Vardenafil Dapoxetine HCl
Standard
Drug
Target
Concentration
(μg/ml)
Peak Area
of Sample
Found
Concentration
(μg/ml)
Mean SD % RSD
Vardenafil
5 686.218 5.09
4.97 0.03 0.60 5 680.072 4.97
5 677.161 4.95
10 1385.69 10.2
10.06 0.12 1.19 10 1351.49 9.97
10 1358.77 10.02
15 2050.46 15.17
15.06 0.11 0.73 15 2019.84 14.94
15 2037.27 15.07
Dapoxetine
HCl
15 1224.19 15.04
14.97 0.07 0.46 15 1213.19 14.90
15 1219.31 14.98
30 2472.38 30.40
29.99 0.37 1.2 30 2411.39 29.65
30 2433.31 29.92
45 3658.94 45.00
44.72 0.34 0.76 45 3604.27 44.33
45 3644.38 44.83
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 26
Interday Precision
The SD and % RSD for Interday precision of
both the drugs was found to be less than 1 and
less than 2 respectively. So, it was concluded
that proposed method for estimation of
Vardenafil and Dapoxetinel HCl is précised in
nature.
Limit of Detection
The proposed method can detect Vardenafil and
Dapoxetine HCl at very low level .So, it was
concluded that the proposed method is very
sensitive in nature.
Limit of Quantitation
The proposed method can quantify small
amount of drugs with precisely. So, it was
concluded that the proposed method is very
sensitive in nature.
Table 11: Interday precision data for Vardenafil and Dapoxetine HCl
Standard
Drug
Target
Concentratio
n (μg/ml)
Peak Area
of Sample
Found
Concentration
(μg/ml)
Mean SD % RSD
Vardenafil
5 681.44 4.97
4.95 0.05 1.01 5 689.63 5.0
5 670.74 4.9
10 1381.58 10.1
10.01 0.07 0.69 10 1352.85 9.97
10 1352.45 9.97
15 2066.81 15.29
15.14 0.15 0.99 15 2025.97 14.98
15 2047.57 15.15
Dapoxetine
HCl
15 1215.66 14.93
14.96 0.13 0.86 15 1230.32 15.11
15 1208.31 14.84
30 2465.09 30.31
29.93 0.33 1.10 30 2413.82 29.68
30 2423.54 29.80
45 3688.13 45.36
44.99 0.46 1.02 45 3615.25 44.47
45 3669.94 45.14
Table 12: LOD data for Vardenafil and Dapoxetine HCl
Parameters Vardenafil Dapoxetine HCl
Mean Slope (n=5) 134.92 81.228
SD of Y-intercept (n=5) 7.684 2.0748
LOD (μg/ml) 0.18 0.10
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 27
Results of forced degradation study by RP-
HPLC:
Acid Hydrolysis
Figure 8: Chromatogram of Acid degradation of
Blank
Figure 9: Chromatogram of standard Drug
Table 13: LOQ data for Vardenafil and Dapoxetine HCl
Parameters Vardenafil Dapoxetine HCl
Mean Slope (n=5) 134.92 81.228
SD of Y-intercept (n=5) 7.684 2.0748
LOQ (μg/ml) 0.56 0.32
Robustness
Table 14: Robustness data for Vardenafil and Dapoxetine HCl
Parameters
(n+5) Variation
Average Peak Area % RSD
Vardenafil Dapoxetine
HCl Vardenafil
Dapoxetine
HCl
Flow rate 1.1 1292.40 2310.41 1.20 1.11
0.9 1424.29 2551.01 1.24 1.02
Mobile phase 56:44 1361.76 2434.14 0.69 0.95
54:46 1359.79 2430.08 1.05 1.00
pH 3.6 1361.56 2431.72 1.23 1.20
3.4 1360.03 2430.00 1.29 1.21
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 28
Figure 10: Chromatogram of Acid degradation
of drug product
Impurities generated from acid degradation
were well separated by this method with good
resolution.
Alkali Hydrolysis
Figure 11: Chromatogram of Alkali degradation
of Blank
Figure 12: Chromatogram of Alkali degradation
of drug product
Impurities generated from alkali degradation
were well separated by this method with good
resolution.
Oxidative Degradation
Figure 13: Chromatogram of Oxidative
degradation of Blank
Figure 14: Chromatogram of Oxidative
degradation of drug product
Impurities generated from oxidative degradation
were well separated by this method with good
resolution.
Thermal Hydrolysis
Figure 15: Chromatogram of Thermal
degradation of drug product
Development and Validation of Analytical Method for Simultaneous Estimation of Vardenafil and Dapoxetine HCl in Bulk and in Combined Dosage Form
© Copyright reserved by IJPRS Impact Factor = 1.0285 29
Impurities generated from Thermal degradation
were well separated by this method with good
resolution.
Photolytic Degradation
Figure 16: Photolytic degradation of drug
product
Impurities generated from Photolytic
degradation were well separated by this method
with good resolution.
CONCLUSION
A simple, specific, accurate and precise RP-
HPLC method has been developed and validated
for simultaneous estimation of Vardenafil and
Dapoxetine HCl in its dosage form. Vardenafil
and Dapoxetine HCl were estimated on
HYPERSIL BDS C18 column using Buffer
0.02m KH2PO4:Acetonitrile (pH 3.5+0.1%TEA)
(55:45 v/v) as mobile phase and detection was
carried out at 266 nm. The linearity range was
found to be 5-15 μg/ml for Vardenafil and 15-45
μg/ml for Dapoxetine HCl. The co-relation
coefficient was found to be 0.998 and 0.999 for
Vardenafil and Dapoxetine HCl respectively.
The assay value for Vardenafil and Dapoxetine
HCl was found to be 100.22% and 99.44%
respectively. A validated stability-indicating
HPLC analytical method has been developed for
the determination of Vardenafil & Dapoxetine
HCl in bulk and in dosage forms. Vardenafil
and Dapoxetine HCl does not undergo any
degradation so it can be said that Vardenafil and
Dapoxetine HCl is stable to forced degradation
conditions.
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