j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 9 4 5 e9 5 3

Available online at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/jopr

Original Article

Stability-indicating HPTLC method for

simultaneous determination of Ketoprofen, Methyl
Paraben and Propyl Paraben in gel formulation
Pallavi Mangesh Patil a,*, Sagar Baliram Wankhede b,
Praveen Digambar Chaudhari c
a

Centre for Research and Development, Prist University, Vallam, Thanjavur, Tamil Nadu 613403, India
Department of Pharmaceutical Chemistry, Padm. Dr. D.Y. Patil Institute of Pharmaceutical Sciences & Research,
Pimpri, Pune 411018, Maharashtra, India
c
Department of Pharmaceutics, P.E. Societys Modern College of Pharmacy, Yamunanagar, Nigdi, Pune 411044,
Maharashtra, India
b

article info

abstract

Article history:

Aim: A novel and quick HPTLC-densitometric method was developed for the simultaneous

Received 20 August 2013

determination of Ketoprofen, Methyl Paraben, and Propyl Paraben.

Accepted 2 September 2013

Methods: Chromatographic separation of the drugs was performed on precoated silica gel 60

Available online 26 September 2013

F254 Merck plates using Toluene:Ethyl acetate:Glacial acetic acid (6.5:2.5:1.0 v/v/v) as a
mobile phase. A TLC scanner set at 265 nm was used of Ketoprofen, Methyl Paraben, Propyl

Keywords:

Paraben respectively were validated according to ICH guidelines. Forced degradation con-

Ketoprofen (KETO)

ditions of hydrolysis (neutral, acidic and alkaline), oxidation, photolysis and thermal stress,

Methyl Paraben (MP)

as suggested in the ICH guideline Q1A (R2).

Propyl Paraben (PP)

Results: The three drugs were satisfactorily resolved with Rf values of 0.33
0.05,

HPTLC

0.54
0.05, 0.71
0.05 for Ketoprofen, Methyl Paraben, Propyl Paraben respectively. Cali-

Force degradation studies

bration curves were polynomial in the range 200e1000 ng/band, 200e1500 ng/band, 100
e600 ng/band, for Ketoprofen, Methyl Paraben, and Propyl Paraben respectively. Correlation coefficient (r) values were 0.9917, 0.9927, 0.9906 Ketoprofen, Methyl Paraben, Propyl
Paraben respectively. The percentage recovery ranges from 99 to 101%.
Conclusion: A low relative standard deviation (<2%) was found for both precision and
robustness study showing that the proposed method was precise and robust. The method
had an accuracy of 99.95%, 99.85% and 100.07 of Ketoprofen, Methyl Paraben, Propyl Paraben respectively were validated according to ICH guidelines. The drug showed instability
in oxide , heat and UV light, while it remained stable in neutral conditions.
Copyright 2013, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights
reserved.

* Corresponding author. Tel.: 91 9823720695 (mobile).

E-mail addresses: pallavipatil_2007@yahoo.com, psadanshio@yahoo.co.in (P.M. Patil).
0974-6943/$ e see front matter Copyright 2013, JPR Solutions; Published by Reed Elsevier India Pvt. Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jopr.2013.09.004

946

1.

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Introduction

A new pharmaceutical preparation (gel) containing ketoprofen (Fig .1) as an active compound with anti-inflammatory and
analgesic activity was developed for treatment of diseases of
the muscolo-skeletal apparatus, in which a local action is
preferred. In order to prevent bacterial growth during the
storage of the formulation,1,2 two commonly used preservativesda mixture of the methyl ester and propyl ester of phydroxybenzoic acid Methyl Paraben (MP) (Fig. 2) and Propyl
Paraben (PP) (Fig. 3)dhave been used gas chromatographyemass spectrometry (GCeMS),3 capillary electro chromatography,4,5 high-performance liquid chromatography (HPLC)
6e8
, HPLCeMS9,10 or micellar chromatography11 as well. Only
one HPLC method has been found in literature12 for simultaneous determination of KP and its degradation products, but
not in the presence of preservatives. Recently, preservatives in
pharmaceuticals have to be quantified. HPLC analysis of MP
and PP is frequently described in the literature13e15; another
publication deals with simultaneous quantification of Ketoprofen and Parabens in a commercial gel formulation by
RPeHPLC with UV detection,16 but there is no any HPTLC
method describing simultaneous determination of all three
componentsdketoprofen, MP and PPdin pharmaceutical
preparations with no any HPTLC method describing simultaneous determination in this mobile phase with beneficial
system suitability parameter.
For such a formulation, a novel method capable to analyze
simultaneously the active component ketoprofen, and its two
preservatives Methyl Paraben and Propyl Paraben was developed. Thereafter, this HPTLC method17 was successfully
applied for the separation, quantification and stability study
of all these compounds in formulated ketoprofen gel 2.5%.

Fig. 1 e Ketoprofen.

Fig. 2 e Methyl Paraben.

2.2.

Preparation of standard stock solutions

2.2.1.

Stock solution A

Accurately weighed quantity (100 mg) of KETO was transferred to 100.0 mL volumetric flask, dissolved and diluted up to
the mark with mobile phase. From this solution, 5.0 mL was
transferred to 50.0 mL volumetric flask and diluted to the
mark with mobile phase (concentration 100 mg/mL). The solution was mixed and filtered through 0.2 m membrane filter.

2.2.2.

Stock solution B

2.

Material and method

Accurately weighed quantity (100 mg) of MP was transferred

to 100.0 mL volumetric flask, dissolved and diluted up to the
mark with mobile phase. From this solution, 5.0 mL was
transferred to 50.0 mL volumetric flask and diluted to the
mark with mobile phase (concentration 100 mg/mL). The solution was mixed and filtered through 0.2 m membrane filter.

2.1.

Reagents and chemicals

2.2.3.

Toluene, Ethyl acetate, Glacial acetic acid from S. D. Fine

Chemicals, Mumbai Reference standard Ketoprofen and
Methyl Paraben and Propyl Paraben were procured from ZIM
laboratories, Nagpur, India as gift samples. Formulated gel
formulation (Ketoprofen 2.5% w/w).
Instrumentation and chromatographic conditions are
given in the following table:

Sr.
no.

Instruments

Descriptions

1
2
3
4
5

HPTLC system
Sample application
Scanner
Software
Saturated chamber

HPTLC plate

Syringe

Camag HPTLC system

Camag Linomat IV automatic sample
Camag TLC scanner
Camag winCATS software
Camag twin-trough chamber (10  10)
and (20  20)
Merck HPTLC plate coated with silica
gel 60 F 254 (0.2 mm thickness) on
aluminum sheet
Hamilton syringe (100 ml)

Stock solution C

Accurately weighed quantity (100 mg) of PP was transferred to

100.0 mL volumetric flask, dissolved and diluted up to the
mark with mobile phase. From this solution, 5.0 mL was
transferred to 50.0 mL volumetric flask and diluted to the
mark with mobile phase (concentration 100 mg/mL). The solution was mixed and filtered through 0.2 m membrane filter.

2.2.4.

Stock solution D

An accurately weighed quantity of 250 mg KETO and 100 mg

MP, 10 mg was transferred to 100.0 mL volumetric flasks,
40.0 mL of mobile phase was added; the content was dissolved
and diluted up to the mark with mobile phase. From this solution, 5.0 mL was transferred to 10.0 mL volumetric flask and
diluted to the mark with mobile phase. Further, 5.0 mL of

Fig. 3 e Propyl Paraben.

j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 9 4 5 e9 5 3

above solution was diluted to 10.0 mL with mobile phase

(concentration of 625 mg/mL KETO and 250 mg/mL MP, 25 mg/mL
PP respectively). The solution was mixed and filtered through
0.2 m membrane filter.

2.3.

947

Migration distance: z80 mm


Temperature: 20
5 C
Scanning mode: Absorbance/Reflectance
Slit dimensions: 5  0.45 mm
Scanning wavelength: 265 nm

Selection of mobile phase

The retention factors of KETO, MP and PP were:

Aliquot portion of standard stock solutions D (5 mL each) was

applied on TLC plates in the form of band (band size: 6 mm).
Different solvents with varying polarity as well as combination of solvent were tried to get well separated bands of the
drugs. After trying several permutations and combinations,
the solvent system containing Toluene:Ethyl acetate:Glacial
acetic acid (6.5:2.5:1.0 v/v/v) was found to be most satisfactory
as it gave good resolution of both drugs.

2.4.
Selection of wavelength for densitometric
evaluation of separated bands
Standard stock solution D was applied on TLC plate with the
help of CAMAG LINOMAT-V automatic sample applicator, the
plate was chromatographed in twin-through glass chamber
saturated with mobile phase for 30 min. After chromatographic development, the plate was removed and air dried.
The separated bands on the TLC plate were scanned over the
wavelength range of 200e700 nm. The wavelength 265 nm
was selected for densitometric evaluation of separated bands.
The overlain spectrum obtained is depicted in Fig. 4.

2.5.

Optimum chromatographic conditions

Stationary phase: Aluminum plates precoated with silica gel

60 F254 (Merck)
Mobile phase: Toluene:Ethyl acetate:Glacial acetic acid
(6.5:2.5:1.0 v/v/v)
Plate size: 10 cm  10 cm
Mode of application: Band
Band size: 6 mm (Distance between two bands: 5.5 mm)
Sample volume: 5 mL
Development chamber: Twin-through glass chamber,
10 cm  10 cm with (20  20) stain less steel lid
Saturation time: 30 min
Separation technique: Ascending

KETO: 0.33
0.05

MP: 0.54
0.05
PP: 0.71
0.05
Densitogram of KETO, MP and PP is shown in Fig. 5.

2.6.

Study of linearity range

The standard stock solution A containing KETO and standard

stock solution B containing MP and stock solution C containing PP was applied on the TLC plate in the range 1e6 mL with
the help of micro syringe using LINOMAT-V automatic sample
applicator. The plate was then developed and scanned under
the above mentioned chromatographic conditions.
Rf was recorded for each drug concentration and the calibration curves of the concentration vs. Rf were constructed for
both the drugs. The calibration curve for KETO and MP and PP
are depicted in Figs. 6e8 respectively.

2.6.1.

Selection of mobile phase

From standard stock D was appropriately to obtain final concentration 625 mg/mL KETO and 250 mg/mL MP, 25 mg/mL PP
respectively. The diluted standard solutions were filtered
through 0.2 m membrane filter. After trying several permutations and combinations, the solvent system containing Toluene:Ethyl acetate:Glacial acetic acid (6.5:2.5:1.0 v/v/v) was
found to be most satisfactory as it gave good resolution of both
drugs.

2.7.

Preparation of gel formulation of Ketoprofen

Ketoprofen gel formulation was prepared using 1% carbopol

940 and as a gelling agent. Gelling agent was dispersed in a
small quantity of distilled water 75 ml and then stored overnight to ensure complete hydration. Ketoprofen in a suitable

Fig. 4 e Overlain spectra of KETO, MP and PP.

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Fig. 5 e Typical densitogram of KETO, MP and PP.

solvent (water) as added to the dispersion and make up weight

with distilled water.
Other excipients (Methyl Paraben 1% and propyl Paraben
0.1%) were also added slowly with continuous stirring. In
carbopol gels, pH of the vehicle was brought to neutral by
using TEA (Triethanolamine).
The final weight of the gel was adjusted to 100 gm with
distilled water. Entrapped air bubbles were removed by
keeping the gels in vacuum desiccators as shown in Table 1.

2.8.

Analysis of gel formulation

An accurately weighed quantity of gel was weighed equivalent

to about 1000 mg of Ketoprofen and 400 mg of Methyl Paraben
and 40 mg Propyl Paraben into a 1000 mL volumetric flask. And
appropriate amount of methanol was then added. The
mixture was ultrasonicated for 30 min with heating and
allowed to cool at room temperature before adjusting to volume with methanol. The organic layer was decanted and the
extraction procedure was repeated.
The resulting mixture was centrifuged at 3500 rpm for
20 min and 20 mL of the clear supernatant was injected directly

Fig. 6 e Standard calibration curve for Ketoprofen.

onto the column. After centrifugation 20 mL of this mixture

was injected into the chromatograph. The resulting solution
was mixed and filtered through Whatman filter paper and
filtrate was appropriately diluted to get approximate concentration and to obtain final concentration of 1000 mg/mL KETO
and 400 mg/mL MP, 40 mg/mL respectively. The diluted solution
was filtered through 0.20 m filter.
On the TLC plate two bands of standard stock solution D
and four bands of sample solution, 5.0 mL each, were applied
and the plate was developed and scanned under the optimum
chromatographic condition. After chromatographic development the peak obtained for standard and sample bands was
integrated. The amount of KETO, MP and PP present in applied
volume of standard solution was fed to computer. Amount of
drug present in applied volume of sample solution was obtained by comparing Rf of sample bands with that of standard
bands.
Amount of drug estimated in mg/gel and the percent label
claim were calculated using the following formula:
The content of KETO, MP and PP in sample was calculated
using the following formula no. 1.

Fig. 7 e Standard calibration curve for Methyl Paraben.

949

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2.9.1.

Accuracy

To as certain the accuracy of proposed method, recovery

studies were carried out by standard addition method, as per
ICH guidelines.

2.10.

Preparation of sample solutions

An accurately weighed quantity of pre-analyzed gel equivalent to about 1000 mg KETO, 400 mg MP and 40 mg PP was
transferred individually in nine different 1000.0 mL volumetric flasks. To each of the flask following quantities of
KETO, MP and PP were added:
Fig. 8 e Standard calibration curve for Propyl Paraben.

Table 1 e Composition of the carbopol and pure drug

Ketoprofen as below.
Ingredient

Quantity taken

Ketoprofen
Methyl Paraben
Propyl Paraben
Carbopol (1%) as gel base
Double distilled water
Triethanolamine

2.5 g
1.0 g
0.1 g
QS
Make up to 100 ml
Q S to neutralize gel

Q.S (Quality sufficient).

Amount of drug estimated mg=gel

Mean amount estimated mg in applied volume

Volume of sample solution applied mL


Volume of stock solution mL

Wt: of gel taken mg

 Average wt: of gel mg

(1)

Percent label claim was calculated using above formula

no 1.
Results of analysis of gel formulation and its statistical
evaluation are shown in Tables 2 and 3 respectively.

2.9.

Method validation

The proposed method was validated by studying several parameters such as accuracy, precision, linearity, limit of
detection (LOD), limit of quantitation (LOQ) and robustness.

Flask no.1: 800 mg KETO 320 mg MP 32 mg PP

Flask no.2: 800 mg KETO 320 mg MP 32 mg PP
Flask no.3: 800 mg KETO 320 mg MP 32 mg PP
Flask no.4: 1000 mg KETO 400 mg MP 40 mg PP
Flask no.5: 1000 mg KETO 400 mg MP 40 mg PP
Flask no.6: 1000 mg KETO 400 mg MP 40 mg PP
Flask no.7: 1200 mg KETO 480 mg MP 48 mg PP
Flask no.8: 1200 mg KETO 480 mg MP 48 mg PP
Flask no.9: 1200 mg KETO 480 mg MP 48 mg PP
Then 100 mL methanol was added to each flask and content of the flask was ultrasonicated for 20 min, volume was
then made up to the mark with mobile phase. The solution
was individually mixed and filtered through Whatman filter
paper no. 42. From the filtrate, 1.0 mL solution was diluted to
10.0 mL with mobile phase. The diluted solution was filtered
through 0.2 m membrane filter.
On the TLC plate two bands of standard stock solution D
and four bands of sample solution, 5.0 mL each, were applied
and the plate was developed and scanned under the optimum
chromatographic condition. After chromatographic development the peak obtained for standard and sample bands were
integrated. The amount of KETO, MP and PP present in applied
volume of standard solution was fed to computer. Amount of
drug present in applied volume of sample solution was obtained by comparing Rf of sample bands with that of standard
bands.
Amount of KETO, MP and PP in sample was calculated by
comparing the mean Rf for standard and sample solution by
formula no. 2.
Amount of KETO, MP and PP in sample (mg) was calculated
by following formula:

Table 2 e Results of analysis gel formulation.

Formulated gel
Sr. no.

1.
2.
3.
4.
5.
6.

Average weight: 4.000 mg

Weight of gel taken (mg)

4000
4000
4000
4000
4000
4000

Amount of drug estimated (mg/Gel taken)

% Label claim

KETO

MP

PP

KETO

MP

PP

1000.09
1000.01
1000.00
999.98
999.78
1000.01

399.00
399.12
400.00
400.12
398.78
401.02

40.00
39.87
39.87
40.00
41.02
41.77

100.90
100.49
100.00
99.88
99.79
100.03

99.80
99.87
100.00
100.12
99.88
100.73

100.00
99.89
99.89
100.00
101.00
101.01

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Table 3 e Statistical validation for analysis of gel formulation.

Sr. no.

Drug

Amount of drug estimated (mg/gel)a

% Label claima

S.D

C.V

S.E

1.
2.
3

KETO
MP
PP

999.99
399.67
40.42

99.95
99.85
100.00

1.3143

1.2108

0.2147

1.4278

1.2146

0.2251

0.3181
0.1504
0.1151

a Mean of six determinations.

Table 4 e Results of recovery studies.

Level of
recover

Weight of gel
taken (mg)

80%

Amount of drug added (mg)

Amount of drug recovered (mg)

KETO

MP

PP

KETO

MP

PP

KETO

MP

PP

800
800
800
1000
1000
1000
1200
1200
1200

320
320
320
400
400
400
480
480
480

32
32
32
40
40
40
48
48
48

799.78
800.01
800.12
999.18
999.70
1000.10
1198.78
1200.00
1201.03

318.13
319.79
320.11
399.18
399.77
400.18
479.11
480.72
480.75

31.99
32.00
32.11
40.10
39.98
40.11
47.91
47.95
48.01

99.71
100.01
100.19
99.78
99.88
100.02
99.93
100.00
101.02

98.11
99.89
100.11
99.77
99.85
100.11
99.10
100.42
100.47

99.98
100.00
100.12
100.11
99.92
100.12
99.92
99.95
100.01

4000
4000
4000
4000
4000
4000
4000
4000
4000

100%

120%

% Recovery

Table 5 e Statistical validation for recovery study.

% Mean recoverya

Level of recovery

80%
100%
120%

Standard deviation

% R.S.D.

S.E

KETO

MP

PP

KETO

MP

PP

KETO

MP

PP

KETO

MP

PP

99.97
99.95
100.02

99.89
99.85
100.15

100.10
100.05
100.15

1.2679

1.2454

1.2788

1.2488

1.2214

1.2958

1.3011

1.3117

1.2481

1.2519
1.2316
1.2668

1.2310
1.2161
1.2848

1.2923
1.3109
1.2316

0.6111
0.5922
0.7155

0.5977
0.5811
0.7066

0.7122
0.7211
0.6811

a Mean of three determinations.

Amount of drug KETO; MP and PP mL estimated mg

2.11.2. Inter-day precision

Mean amount estimated mg in applied volume

Volume of sample solution applied mL

 Volume of stock solution

(2)

Amount of the drug recovered (mg) and % recovery was

calculated and results of recovery studies and statistically are
shown in Tables 4 and 5.

2.11.

Inter-day precision was determined by analyzing Gel sample

solutions on three different days. Gel sample solution was
prepared and analyzed in the similar manner as described in
analysis of the gel formulation.
Results of intra-day precision and inter-day precision are
shown in Tables 6 and 7, respectively.

2.12.
(LOQ)

Precision

2.11.1. Intra-day precision

Intra-day precision was determined by analyzing Gel sample
solutions at different time intervals on the same day. Gel
sample solution was prepared and analyzed in the similar
manner as described under analysis of the gel formulation.

Table 6 e Intra-day precision data.

a

Limit of detection (LOD) and limit of quantitation

The LOD and LOQ were separately determined which is based

on the standard deviation of response of the calibration curve.
The standard deviation of y-intercept and slope of the calibration curves were used to calculate the LOD and LOQ. Results are shown in Table 8.

Table 7 e Inter-day precision data.

Drug

% Mean

S. D.

C. V.

Drug

% Meana

S.D.

C.V.

KETO
MP
PP

99.91
99.94
99.89

0.3110

0.2981

0.5912

0.3212
0.2012

0.6073

KETO
MP
PP

99.15
100.02
99.65

1.7061

0.4173

1.3012

1.7170
0.4153

1.3007

a Average of six determinations.

a Average of six determinations.

j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 9 4 5 e9 5 3

2.13.

Table 8 e LOD and LOQ.

Parameter

KETO

MP

PP

Limit of detection (ng/band)

Limit of quantification (ng/band)

138.41
418.15

58.15
108.14

24.16
68.15

Table 9 e Result of robustness study.

951

Robustness of method

To evaluate the robustness of the proposed method, small but

deliberate variations in the optimized method parameters
were done. The effect of change in flow rate and mobile phase
ratio on retention time and tailing factor were studied. The
solution containing 25 mg/mL of KETO, 12.5 mg/mL of MP and
0.5 mg/mL of PP was injected (in triplicate) into sample injector
of HPLC three times under the varied conditions. Robustness
data is given in Table 9.

Chromatographic changes
Factor
Mobile phase composition
(
0.1 mL)
2.4:3.4:3.9
2.5:3.5:4
2.6:3.6:4.1
Amount of mobile phase (v/v)
(
1 mL)
9
10
11

Level

e0.1
0
0.1

e1.0
0
1.0

Rf value
KETO

MP

PP

0.32
0.34
0.35

0.64
0.65
0.68

0.70
0.71
0.74

KETO

MP

PP

0.33
0.34
0.36

0.65
0.65
0.67

0.71
0.71
0.75

3.

Forced degradation study of ketoprofen

Amount of gel equivalent to about 25 mg KETO was separately

transferred to five different 25.0 mL volumetric flasks (Flask
no. 1, 2, 3, 4 and 5), added 5.0 mL of 0.1 M HCl, 0.1 M NaOH and
3% H2O2 to Flask no. 1, 2 and 3, respectively. Solution in flask
no. 1, 2, and 3 were heated in water bath for 3 h at 80  C. Flask
no. 4 containing gel was kept at 60  C for 24 h to study the
effect of heat on Gel sample (heat degradation). The forced
degradation was performed in the dark to exclude the possible
degradative effect of light. Flask no. 5 was exposed to ultraviolet radiations at 254 nm for 24 h in a UV-chamber. All the

Fig. 9 e Chromatogram of 0.1 M NaOH gel formulation.

Fig. 10 e Chromatogram of 0.1 M HCl gel formulation.

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Fig. 11 e Chromatogram of H2O2 (3%) gel formulation.

Fig. 12 e Chromatogram of dry heat gel formulation.

flasks were removed Gel samples were treated and analyzed
in similar manner as described under analysis of gel
formulation.
The typical densitogram is shown in Figs. 9e13for acidic,
alkaline, oxide, heat and UV exposure, respectively.
Results of forced (stress) degradation studies are shown in
Table 10.

4.

Results and discussion

In the present work, new method namely, simultaneous

equation method and quick high-performance liquid

chromatography (HPLC) method were developed and validated

for the simultaneous determination of three compounds in a
formulated gel. The three drugs were satisfactorily resolved
with Rf values of 0.33
0.05, 0.54
0.05, 0.71
0.05 for
Ketoprofen, Methyl Paraben, Propyl Paraben respectively.
Calibration curves were polynomial in the range 200e1000 ng/
band, 200e1500 ng/band, 100e600 ng/band, for Ketoprofen,
Methyl Paraben, and Propyl Paraben respectively. Correlation
coefficient (r) values were 0.9917, 0.9927, 0.9906 Ketoprofen,
Methyl Paraben, Propyl Paraben respectively. A low relative
standard deviation (<2%) was found for both precision and
robustness study showing that the proposed method was precise and robust. The method had an accuracy of 99.96%, 99.91%

Fig. 13 e Chromatogram of UV radiation gel formulation.

j o u r n a l o f p h a r m a c y r e s e a r c h 6 ( 2 0 1 3 ) 9 4 5 e9 5 3

Table 10 e Results of degradation study.

Sr. Stress condition Percent assay of
no.
active substance
(ketoprofen)
1.
2.
3.
4.

Acid (0.1 M HCl)

Alkali (0.1 M NaOH)
Oxide (3% H2O2)
Heat (60  C)

99.65
99.53
97.76
97.82

5.

UV (240 nm)

98.69

Acknowledgment
Rf value of
degraded
product

0.33, 0.49, 0.67

0.33, 0.72
0.12, 0.34, 0.50, 0.64
0.34, 0.51, 0.64, 0.66,
0.78
0.33,0.58,0.77

and 101.05 Ketoprofen, Methyl Paraben, Propyl Paraben

respectively. Method had the potential to determine these
drugs simultaneously from dosage forms without any interference, in accordance with ICH guidelines. The limit of
detection was 138.41 ng/band, 58.15 ng/band and 24.16 ng/band
for KETO, MP and PP respectively and limit of quantification
was 418.15 ng/band, 108.14 ng/band and 68.15 ng/band for
KETO, MP and PP respectively and the method was found to be
specific. The percentage recovery ranges from 99 to 101%.
Forced degradation conditions of hydrolysis (neutral, acidic and
alkaline), oxidation, photolysis and thermal stress, as suggested in the ICH guideline Q1A (R2). The drug showed instability in acid and oxide, while it remained stable in neutral
conditions.

5.

Conclusion

The proposed method for simultaneous estimation (HPLC) of

Ketoprofen, Methyl Paraben and Propyl Paraben in their
formulated gel dosage and validated as per ICH guidelines.
Moreover the method is economic, simple and rapid, hence
can be employed for routine analysis in quality control
laboratories.

Conflicts of interest
All authors have none to declare.

953

I sincerely thank Zim Laboratory, Nagpur, Maharashtra and

Gen Pharmaceuticals, Pune, Maharashtra for providing me the
gift sample of KETO, MP and PP and I thank my lab technicians
for their contribution.

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