Ruchipriya Jogarami et al.

/ Journal of Pharmacy Research 2012,5(4),2273-2275

Research Article Available online through
ISSN: 0974-6943 www.jpronline.info
Validated UV Spectrophotometric Method Development for Simultaneous
Estimation of Tazarotene and Hydroquinone in Gel Preparation
Ruchipriya Jogarami 1 *, Dr. Prateek Jain1, Sheetal Sharma2
Adina Institute of Tech. & Science, Sagar, Madhya Pradesh, India
Received on:11-01-2012; Revised on: 17-02-2012; Accepted on:19-04-2012

ABSTRACT
Simple, accurate, and economical procedures for simultaneous estimation of Tazarotene and Hydroquinone in gel form have been developed. The method
employs formation and solving of simultaneous equation using 247.2 nm and 289.6 nm as two analytical wavelengths for both drugs in methanol. Tazarotene
and Hydroquinone at their respective λmax 247.2 nm and 289.6 nm shows linearity in a concentration range of 5-25 µg/ml and 5-25 µg/ml respectively.

Key words: Tazarotene, Hydroquinone, λmax, Simultaneous equation method.

INTRODUCTION:
Tazarotene [(Ethyl 6-[2-(4,4-dimethylthiochroman-6-yl) ethynyl] nicoti-
nate (Fig. 1a) is a well known synthetic acetylenic retinoid, that is applied
topically as cream or gel. It is de-esterified in the skin to its active form,
tazarotene acid, which affects cell proliferation and differentiation by modu-
lating gene expression in acne and psoriasis. This medication is approved for
treatment of psoriasis, acne, and sun damaged skin (photodamage).
Hydroquinone is Quinol/1-4 dihydroxy benzene/1-4 hydroxy benzene (Fig.
1b). It acts by inhibiting Hydroquinone has a variety of uses principally
associated with its action as a reducing agent which is soluble in water. It is a
major component in most photographic developers for film and paper where,
with the compound Metol, it reduces silver halides to elemental silver.
A literature review reveals that no analytical method is available for the
simultaneous estimation of Tazarotene and Hydroquinone in gel form in
pharmaceutical preparations, which prompted to pursue the present work.
The objective of the present work is to develop and validate new analytical
methods for simultaneous determination of Tazarotene and Hydroquinone in
gel form.
UV/VIS Spectrophotometer with 1cm. matched quartz cells. Glasswares
used in each procedure were soaked overnight in a mixture of chromic acid
and sulphuric acid rinsed thoroughly with double distilled water and dried in
hot air oven. Tazarotene and Hydroquinone reference standard were pro-
cured from Macleod’s Pharmaceutical Ltd. The pharmaceutical preparation
of combination of Tazarotene and Hydroquinone is prepared Sapiens phar-
maceutical lab, Bhopal. Methanol of analytical grade was purchased from
Merck & Co. All the solutions were protected for light and were analyzed on
the day of preparations.
Selection of common solvent:
Methanol of analytical reagent grade was selected as common solvent for
developing spectral characteristics of drug. The selection was made after
assessing the solubility of both the drugs in different solvents.

Fig.2a Selection of λmax of Tazarotene

Fig.1a Fig.1b
MATERIALS AND METHODS:

Materials:
Spectral runs were made on Thermospectronic model of Elico India SL-159

*Corresponding author.
Ruchipriya Jogarami
Adina Institute of Tech. & Science,
Sagar, Madhya Pradesh, India

Fig.2b Selection of λmax of Hydroquinone

Journal of Pharmacy Research Vol.5 Issue 4.April 2012 2273-2275
 Ruchipriya Jogarami et al. / Journal of Pharmacy Research 2012,5(4),2273-2275
Table 2: Results of analysis of tablet samples.
Preparation of Standard Stock Solution:
10 mg of Tazarotene was weighed accurately and transferred to a 10ml Tazarotene Hydroquinone
Label Claim (mg) % Purity* Label Claim (mg) % Purity*
volumetric flask, and the volume was adjusted to the mark with the diluent
(Methanol), to give a stock solution of 1000ppm and 10 mg of Hydro- 0.1 % 99.6 4% 99
quinone was weighed accurately and transferred to a 10ml volumetric flask,
*Denotes average of five determinations
and the volume was adjusted to the mark with the diluent ( Methanol) to give
a stock solution of 1000ppm Method Validation:
The method was validated for accuracy, precision, specificity, and robust-
Procedure: ness by the following procedures.
Determination of Absorption Maxima:
Standard solution (10µg/ml) of pure Tazarotene and Hydroquinone was Accuracy: The accuracy of the method was determined by calculating recov-
eries of Tazarotene and Hydroquinone by the methods of standard addi-
prepared. The pure drug solutions were scanned on UV spectrophotometer,
tions. This study was performed by addition of known amounts of Tazarotene
which showed maximum absorbance at 347.0 and 289.6 for Tazarotene and
and Hydroquinone to a known concentration of the gel. The amount of
Hydroquinone respectively.
standard recovered was calculated in the terms of mean recovery with the
upper and lower limits of percent relative standard deviation.
Simultaneous equation method:
Two wavelengths selected for the method are 247.2 nm and 289.6 nm that are
Precision: The experiments were repeated for three times a day for intraday
absorption maximas of Tazarotene and Hydroquinone respectively in metha-
precision and on three different days for inter day precision. The developed
nol. The stock solutions of both the drugs were further diluted separately
method was found to be precise for intraday and inter day precision on the
with methanol to get a series of standard solutions of 5-25 µg/ml concentra-
basis of % RSD values for Tazarotene and Hydroquinone.
tions of both drugs. The absorbances were measured at the selected wave-
lengths and absorptivities (A 1%, 1 cm) for both the drugs at both wave- Table 3: Statistical Validation of Recovery Studies
lengths were determined as mean of three independent determinations. Con-
Level of Drug % Recovery Standard % RSD
centrations in the sample were obtained by using following equations- Recovery (%) Deviation*

Cx = A1 ay2 – A2 ay1..................................Eq. (i) 80 TAZA 99.96 0.450 0.451

HYDRO 99.37 0.625 0.628
ax1 ay2 – ax2 ay1
100 TAZA 100.3 0.624 0.622
Cy = A1 ax2 – A2 ax1..................................Eq. (ii) HYDRO 100 1.000 1.000
ay1 ax2 – ay2 ax1 120 TAZA 99.88 0.196 0.196
HYDRO 99.97 0.455 0.455
Where, A1 and A2 are absorbances of mixture at 247.2 nm and 289.6 nm
respectively, ax1 and ax2 are absorptivities of Tazarotene at λ1 and λ2
respectively and ay1 and ay2 are absorptivities of Hydroquinone at λ1 and Table 4: Intra-day and Inter-day precision
λ2 respectively. Cx and Cy are concentrations of Tazarotene and hydro- Intra- day Precision Inter-day Precision
Parameters TAZA % HYDRO % TAZA % HYDRO %
quinone respectively. Label Claim Label Claim Label Claim Label Claim

Application of the proposed method for the determination of Tazarotene Mean 100.28 98.68 98.43 96.80
and Hydroquinone in gel preparation: S.D. 0.655 0.800 1.137 1.609
Gel equivalent to 10 mg of Tazarotene and 10 mg of Hydroquinone was % RSD 0.653 0.810 1.155 1.662
weighed and transferred to a separate 10 ml volumetric flask and volume was
made up to 10 ml with mobile phase to obtain concentration of 1000µg/ml. RESULTS AND DISCUSSION:
Resultant solution was filtered through Whatmann filter paper. 1 ml of fil- The overlain spectra of Tazarotene and Hydroquinone exhibit λmax of 247.2
trate was taken in 10 ml volumetric flask and volume was made up to 10 ml nm and 289.6 nm for Tazarotene and Hydroquinone respectively which are
with mobile phase to obtain concentration of 100µg/ml. Further 1 ml of this quite separated from each other. This wavelength was selected for simulta-
solution was taken and diluted up to 10 ml obtain final concentration of neous estimation of Tazarotene and Hydroquinone and it is assume to be
10µg/ml. Absorbance of the sample solutions at 347.0 nm and 289.6 nm was sensitive wavelength. Standard calibration curves for Tazarotene and Hydro-
measured and from the absorbance values, the concentration of drugs in the quinone were linear with correlation coefficients (r) values in the range of
sample solution was determined by using Vierodt’s formula. 0.986- 0.998 at all the selected wavelengths and the values were average of
three readings with standard deviation in the range of 0.002 – 0.003. The
Table 1: Linear regression analysis of calibration curves with their method was repeated for three different days and average % RSD was found
respective absorptivity values. to be 0.36 for Tazarotene and 0.32 for Hydroquinone.
Parameters TAZA HYDRO
CONCLUSIONS:
Beer’s law limit (µg/ ml) 5-25 (µg/ ml) 5-25 (µg/ ml) The developed spectrophotometric method is simple, accurate, and reliable
Correlation coefficient (r) 0.989 0.998 for the simultaneous estimation of Tazarotene and Hydroquinone in com-
Molar absorptivity (lit/mole/cm) 2.26 X 103 3.85 x 104 bined gel form. The relative std. deviation (RSD) for all parameters was
Sandell’s sensitivity (mcg/Sq.cm/0.001) 0.64 X 10-4 0.35 x 10-4
Slope 0.0387 0.0326 found to be less than one, which indicates the validity of method and assay
Intercept 0.1694 0.0153 results are also within the limit so the proposed method can be used for
routine quantitative simultaneous estimation of both the drugs in multi-
component pharmaceutical preparation.

Journal of Pharmacy Research Vol.5 Issue 4.April 2012 2273-2275

 Ruchipriya Jogarami et al. / Journal of Pharmacy Research 2012,5(4),2273-2275
ACKNOWLDGEMENT:
The author thanks to Macleod’s Pharmaceutical Ltd.for supplying samples
of Tazarotene and Hydroquinone.
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Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.5 Issue 4.April 2012 2273-2275