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Abstract:
This review article gives knowledge of about HPTLC-based analytical method development and validation parameter in accordance to practical evaluation. It meets standards and minimizes errors and investigation. This review article helps to choose best mobile phase and gives guidelines for the good validation practice and understand the steps of analytical procedure.
Keywords: method development, validation, HPTLC
INTRODUCTION:
High Performance Thin Layer Chromatography (HPTLC) is the most powerful advanced form of Thin Layer Chromatography (TLC) and consists of chromatographic layers of utmost separation efficiency and the application of sophisticated instrumentation for all steps in the procedure include accurate sample application, standardized reproducible chromatogram development and software controlled evaluation[1]. HPTLC is a concept that includes a widely standardized methodology based on scientific facts as well as the use of validated methods for qualitative and quantitative analysis [2]. HPTLC meets all quality requirements for today's analytical labs, to increase the resolution and to allow more accurate quantitative measurements [3].
HPTLC method:
Stationary Phase:
HPTLC is the most advanced form of modern TLC. It uses HPTLC plates featuring small particles with a narrow size distribution which results in homogenous layers with a smooth surface to be obtained. HPTLC uses smaller plates (10 x 10 or 10 x 20 cm). HPTLC plates provide improved resolution, higher detection sensitivity, and improved in situ quantification and are used for industrial pharmaceutical densitometric quantitative analysis. Normal phase adsorption TLC on silica gel with a less polar mobile phase, such as chloroform- methanol, has been used for more than 90% of reported analysis of pharmaceuticals and drugs[4].
1.Simple and precise HPTLC methods were developed for the simultaneous estimation of two anti-inflammatory drugs (curcuminand galangin). The method was tailored to analyze both drugs in their commercial dosage form (capsules) with no interference fromingredients. Chromatographic separation was performed over precoated TLC plates (60 F254, 20 cm x 10 cm, 250pm thickness,Merck, Darmstadt, Germany) via a linear ascending technique using n-hexane, ethyl acetate, acetic acid, and methanol as the mobilephase. Detection and quantification was achieved at 404 nm through spectrodensitometricanalysis[5].
2. The report of TLC densitometric method, which has been developed and validated for quantification of stigmasterol from petroleum ether extract of leaves and stems of Bryophyllumpinnatum. The separation was performed on TLC aluminum plates...