*Corresponding Author Address: Manisha S. Phoujdar, Department Quality Assurance Techniques, STES’s, Sinhgad College of
Pharmacy, Vadgaon (Bk.), Pune- 411041, Maharashtra, India; Email- mphoujdar@hotmail.com
World Journal of Pharmaceutical Sciences
ISSN (Print): 2321-3310; ISSN (Online): 2321-3086
Published by Atom and Cell Publishers © All Rights Reserved
Available online at: http://www.wjpsonline.org/
Original Article
Development and validation of HPTLC method for determination of betamethasone
valerate in API and pharmaceutical dosage form
Neha Sharad Sinnarkar and Manisha S. Phoujdar*
Department Quality Assurance Techniques, STES’s, Sinhgad College of Pharmacy, Vadgaon (Bk.), Pune-
411041, Maharashtra, India.
Received: 19-03-2017 / Revised: 24-04-2017 / Accepted: 26-04-2017 / Published: 27-04-2017
ABSTRACT
The aim of present work was to develop a simple and sensitive, HPTLC for the quantitative estimation of
Betamethasone Valerate in its single component cream formulations (20 g). Betamethasone Valerate was
chromatographed on silica Gel 60 F254 TLC plate using Ethyl Acetate: n- Heptane: Toluene: Ethanol
(5.1:2.4:1.2:0.3 v/v/v/v) as mobile phase. Betamethasone Valerate in methanol scanned by Camag TLC scanner
4 with UV visible detector over wavelength range 200 to 400 nm, showed Rf value of 0.34 at wavelength 246
nm and selected for further studies. The method was validated in terms of linearity (1-9 μg/ml), precision (intra-
day variation 1.40, inter-day variation 1.71), accuracy (84 to 96%) and specificity. The limit of detection and
limit of quantification for Betamethasone Valerate were found to be 0.97 μg/spot and 2.96 μg/spot, respectively .
It can be concluded from the results that the proposed method was accurate, precise and consistent the
determination of Betamethasone Valerate in dosage form. This method was validated as per ICH guideline Q2
(R1). Results suggest that this method can be used for routine estimation of Betamethasone Valerate in bulk and
pharmaceutical dosage forms.
Key Words: Betamethasone Valerate, Toluene, Ethanol, n-heptane, Ethyl acetate, HPTLC.
INTRODUCTION
Betamethasone Valerate is a corticosteroid. It
works by modifying the body's immune response to
various conditions and decreasing inflammation.
Betamethasone Valerate is official in Indian
Pharmacopoeia. Literature survey reveals that some
methods have been developed for their
determination by HPLC, HPTLC or
spectrophotometry either alone or in combination.
However, overall cost of analysis of reported
HPTLC method is more. In this view, an
economical HPTLC method has been developed
for estimation of Bethamethasone Valerate in
pharmaceutical dosage form.
MATERIALS AND METHODS
Bethamethasone Valerate standard was provided by
GSK pharmaceuticals, Nasik, India. “Betnovate 20
g” skin cream were procured from local market.
AR grade of solvents used for this study were
purchased from Merck Pvt. Ltd, Mumbai.
Preparation of standard solution: A standard
stock solution of Betamethasone Valerate was
prepared by dissolving 10 mg of standard API in 10
ml of methanol to get concentration of 1000 μg/ml.
This solution was further diluted to get 100 μg/ml
solution of Bethamethasone Valerate as working
standard.
Fig. 1: Chemical Structure of Bethamethasone
Valerate
Selection of wavelength for Detection: The
working standard of Bethamethasone Valerate in
methanol was scanned by Camag TLC scanner 4
with UV-visible detector over wavelength range
200 to 400 nm. Wavelength 246 nm was selected
for further studies. (Figure 2).