Specific High-performance Thin-layer Chromatography Research Articles

A developed high-performance thin-layer chromatography method for the determination of baicalin in Oroxylum indicum L. and its antioxidant activity

A simple, accurate, precise, and specific high-performance thin-layer chromatography (HPTLC) method for the quantitative determination and validation of baicalin in different extracts of Oroxylum indicum has been developed for the first time. The mobile phase of acetone‒ethyl acetate‒water‒formic acid (2:10:0.5:0.5, V/V) was used for achieving good separation. Densitometric determination was carried out at 318 nm. The calibration curves were found to be linear in the range between 200 and 1000 ng per spot. During the analysis, the ethanolic extract of O. indicum showed higher content of baicalin than acetone, DMSO, and DMF extracts. Further, the antioxidant potential of different extracts of O. indicum were assessed with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The developed method of HPTLC was validated for specificity, accuracy, precision and linearity. The ethanolic extract has unveiled significant antioxidant activity with a percentage inhibition of 67.34%.Graphic

STABILITY-INDICATING HPTLC METHOD FOR THE DETERMINATION OF FEBUXOSTAT IN BULK AND PHARMACEUTICAL FORMULATION

Febuxostat (FEB) is a well-known xanthine oxidase (XO) inhibitor that is preferably employed for treating hyperuricemia (extreme stages of uric acid in the human serum). In all the above, particularly, the high-performance thin-layer chromatography (HPTLC) method where the retention factor (Rf) values were found to be quite varying as well as no specific degradation have been yet studied under moisture, sunlight, oxidative stress, acidic environment, and alkaline conditions. The present study exclusively focuses on a much optimized stability-indicating HPTLC-based precise, accurate, and specific HPTLC method for FEB detection in the existence of its degradation components using densitometric detection. The study hereby opened a new perspective in developing a novel HPTLC method for the chromatographic determination of United States Food and Drug Administration (USFDA)-approved drug FEB in bulk and tablet formulations. The method was properly authenticated by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guideline Q2A and guideline Q2B and therefore is found to be accurate, linear, reproducible, precise, robust and economically adequate to execute day after day custom analysis in the pharmaceutical industry scale. The investigation also unties new opportunities for the coherent optimization of HPTLC-based validated analytical methods for other drug products alone as well as simultaneously for frequently available formulations.

Stability indicating high-performance thin layer chromatography method for estimation of ascorbic acid in Hibiscus sabdariffa L. aqueous extract

Background: High performance thin layer chromatography (HPTLC) has become a routine analysis technique due to its advantages of low operating cost, high sample throughput and need minimum sample clean up. Aim: To develop and validate an accurate, specific, repeatable and stability-indicating HPTLC method for the determination of ascorbic acid in presence of their degradation products for assessment of stability of the ascorbic acid in Hibiscus sabdariffa L. preparations. Methods: Chromatographic sample separations employed thin layer chromatography plates (20 cm x10 cm) pre-coated with silica gel 60 F254 as a stationary phase. Solvent system consisted of acetone: toluene: water: glacial acetic acid (26:4:4:1 v/v) while scanning wavelength of 269 nm was used. Ascorbic acid was subjected to stress degradation conditions i.e. oxidation, dry heat treatment, photo degradation and hydrolysis under different PH. Method was validated according to international council for harmonisation guidelines. Results: Peaks of products formed were well resolved with different Rf values. The linear regression data for the calibration plots showed good linear relationship with r2 = 0.9912 in the concentration range 2-4 µg/ml. The mean value slope and intercept were -531.72 and 8040.22 respectively. Conclusion: A rapid, simple, accurate and specific HPTLC method for determination of ascorbic acid in Hibiscus sabdariffa L. preparations was developed and validated. In addition, this method separated all the degradants from stress conditions showing that it is a stability indicating method. The developed method could now be used in quality control of H. sabdariffa herb products.

Standardization of Biomarkers Gallic Acid and Berberine in Polyherbal Formulation Entoban Capsules by High-Performance Thin-Layer Chromatography—Densitometry

The technological improvement in the structural elucidation of natural compounds has made it probable to generate appropriate strategies for the analysis and standardization of plant-based medicines. An appliance of highly oriented hyphenated techniques provides a definite tool for herbal investigations. Therefore, the present study was directed towards the standardization of biomarkers gallic acid and berberine in polyherbal formulation Entoban capsules to ensure the quality of the herbal drugs. A rapid, simple, accurate, and specific high-performance thin-layer chromatography (HPTLC) method for the quantitative estimation of biomarkers berberine and gallic acid has been developed. HPTLC was performed to evaluate the presence of gallic acid and berberine applying toulene—ethyl acetate—formic acid—methanol (12:9:4:0.5 v/v) and ethanol—water—formic acid (90:9:1 v/v), as the mobile phase, respectively. The RF values (0.58 for gallic acid and 0.76 for berberine) in both sample and reference standard were fou...

A Specific High-Performance Thin-Layer Chromatography with Fluorescence Detection for the Determination of Some α1-Blockers

A specific high-performance thin-layer chromatography (HPTLC) method with fluorescence detection was developed for determination of selective α1-blockers, including terazosin (TER), doxazosin, prazosin, and alfuzosin. So far, this is the first HPTLC method using fluorescence detection mode for the determination of α1-blockers. It was designated to combine the specificity attributed to the drug's native fluorescence, the high sample throughputs of the HPTLC methods, and the use of environmentally safe reagents. Good resolution was obtained using simple mobile phase composed of acetonitrile and phosphate buffer (0.02 mole/L) adjusted to pH 3.0 (30:70, v/v). The method used fluorescence detection mode and was validated according to the International Conference on Harmonization guidelines. The emission intensity was measured using optical filter K400 after excitation at 332 nm. The correlation coefficient in the range 0.9984–0.9997 was obtained in the concentration range 5–100 ng/band. The limits of detection and quantification were 3.52–6.91 and 11.76–23.02 ng/band, respectively. Forced degradation stability for TER was studied using the developed HPTLC method. The proposed method was successfully applied for the determination of the studied α1-blockers in pharmaceutical dosage forms and in human plasma without the need for complicated extraction and cleanup steps.