Design of Experiment Approach for the Development and Validation of a Stability-Indicating High-Performance Thin-Layer Chromatography Method for the Estimation of Metformin Hydrochloride and Ursodeoxycholic Acid in Pharmaceutical Dosage Form

Rajendra K. Patel; Vihari K. Shah

doi:10.1556/1006.2019.32.6.6

JPC - Journal of Planar Chromatography - Modern TLC

Volume/Issue:: Volume 32: Issue 6

Design of Experiment Approach for the Development and Validation of a Stability-Indicating High-Performance Thin-Layer Chromatography Method for the Estimation of Metformin Hydrochloride and Ursodeoxycholic Acid in Pharmaceutical Dosage Form

Authors:

Rajendra K. Patel

Rajendra K. Patel Pharmaceutical Chemistry Department, K.B. Institute of Pharmaceutical Education and Research, Gandhinagar, Gujarat 382023, India

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rajukpatel2006@gmail.com

and

Vihari K. Shah

Vihari K. Shah Pharmaceutical Chemistry Department, K.B. Institute of Pharmaceutical Education and Research, Gandhinagar, Gujarat 382023, India

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Pages:: 481–493

Online Publication Date:: Dec 2019

Publication Date:: 01 Dec 2019

Article Category:: Research Article

DOI:: https://doi.org/10.1556/1006.2019.32.6.6

Keywords:: Metformin hydrochloride; Ursodeoxycholic acid; Box-Behnken design; Design of experiment

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Summary

In the present study, a design of experiment (DoE) approach was used to optimize chromatographic conditions for the development of a high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of metformin hydrochloride (MET) and ursodeoxycholic acid (URSO) in pharmaceutical dosage form. The critical factors were identified using a Taguchi design, and after identification of critical factors, optimization was done using Box-Behnken design (BBD). BBD was used to optimize the compositional parameters and to evaluate the main effect, interaction effects, and quadratic effects of the mobile phase compositions, development distance, and saturation time on the retardation factor (R_F) of both drugs. HPTLC separation was performed on aluminum plates pre-coated with silica gel 60 F₂₅₄ as the stationary phase, using toluene–ethanol–acetone–formic acid (4.5:2:2.5:0.85, V/V) as the mobile phase at a wavelength of 234 nm and 700 nm for MET and URSO, respectively. A sharp and well-resolved peak was obtained for MET and URSO at R_F values of 0.19 and 0.80 min, respectively. The calibration curve was in the range of 5000–40000 ng per spot and 1500–12000 ng per spot for MET and URSO, with r² = 0.984 and r² = 0.980, respectively. The method was validated for linearity, accuracy, precision, limit of detection, limit of quantification, and specificity. To provide a better visualization of the statistically significant factors derived from the statistical analysis, the perturbation plot and response surface plot for the effect of independent variables on the R_F of MET and URSO were evaluated. Stability study was performed under different stress conditions such as acid and alkali hydrolysis, oxidation, and temperature. The developed method was able to resolve drugs and their degradation products formed under the afore-mentioned conditions.

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JPC - Journal of Planar Chromatography - Modern TLC

Print ISSN:: 0933-4173

Online ISSN:: 1789-0993

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Journal of Planar Chromatography - Modern TLC
Language	English
Size	A4
Year of Foundation	1988
Volumes per Year	1
Issues per Year	6
Founder	Akadémiai Kiadó
Founder's Address	H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Publisher	Akadémiai Kiadó Springer Nature Switzerland AG
Publisher's Address	H-1117 Budapest, Hungary 1516 Budapest, PO Box 245. CH-6330 Cham, Switzerland Gewerbestrasse 11.
Responsible Publisher	Chief Executive Officer, Akadémiai Kiadó
ISSN	0933-4173 (Print)
ISSN	1789-0993 (Online)