Asian Journal of Research in Chemistry

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Development and Validation of Stability Indicating HPTLC Method for Estimation of Ursolic Acid


     

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A stability indicating high performance thin layer chromatographic (HPTLC) method was developed and validated for estimation of ursolic acid. Study was performed on pre-coated silica gel TLC plates using toluene: ethyl acetate: formic acid (7:3:0.1, v/v/v) as the mobile phase. After development, plates were derivatized with Anisaldehyde- Sulphuric acid reagent. Detection and quantification were performed by densitometric scanning at 530 nm. Method was validated according to ICH Q2(R<SUB>1</SUB>)guideline. The linear regression analysis for the calibration plot showed good linear relationship over the concentration range of 200-600 ng/spot. The correlation coefficient of calibration curve was found to be 0.9986. The intra and inter day precision was found to be less than 2%. Ursolic acid was also subjected to acid, base, oxidation, heat, humidity and photo-degradation studies. The degradation products obtained were well resolved from the pure drug with significantly different Rf values. As the method could effectively separate the drugs from its degradation products, it can be used for stability-indicating analysis.

Keywords

Ursolic Acid, Stability Indicating Method, High Performance Thin Layer Chromatography (HPTLC), Method Validation
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  • Development and Validation of Stability Indicating HPTLC Method for Estimation of Ursolic Acid

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Abstract


A stability indicating high performance thin layer chromatographic (HPTLC) method was developed and validated for estimation of ursolic acid. Study was performed on pre-coated silica gel TLC plates using toluene: ethyl acetate: formic acid (7:3:0.1, v/v/v) as the mobile phase. After development, plates were derivatized with Anisaldehyde- Sulphuric acid reagent. Detection and quantification were performed by densitometric scanning at 530 nm. Method was validated according to ICH Q2(R<SUB>1</SUB>)guideline. The linear regression analysis for the calibration plot showed good linear relationship over the concentration range of 200-600 ng/spot. The correlation coefficient of calibration curve was found to be 0.9986. The intra and inter day precision was found to be less than 2%. Ursolic acid was also subjected to acid, base, oxidation, heat, humidity and photo-degradation studies. The degradation products obtained were well resolved from the pure drug with significantly different Rf values. As the method could effectively separate the drugs from its degradation products, it can be used for stability-indicating analysis.

Keywords


Ursolic Acid, Stability Indicating Method, High Performance Thin Layer Chromatography (HPTLC), Method Validation

References