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Contents:
  1. Account Options
  2. Thin–layer Chromatography (TLC)
  3. INTRODUCTION
  4. Thin-Layer Chromatography: Reagents and Detection Methods - PDF Drive

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Thin Layer Chromatography - Principle

Volume 1b, Physical and chemical detection methods : activation reactions, reagent sequences, reagents II'. Displaying Editions 41 - 46 out of The relative standard deviation RSD of the calculated R f values was close to zero, and the RSD of the raw volumes pixel intensities were 4. This good level of precision was suitable for quality control analysis of AMD.

The error did not exceed 0. Robustness and ruggedness. In order to measure the extent of the method robustness, the most critical parameters were interchanged while keeping the other parameters unchanged, and in parallel the chromatographic profile was observed and recorded. The studied parameters were: the composition of the mobile phase, amount of spraying reagent, and color development time. The results indicated that the small change in the composition of the mobile phase did not significantly affect the R f value of AMD. Both plates yielded the same resolution efficiency and R f values; however, the thinner-layer plate required a longer time for the solvent to reach the marked front.

A compact spots without tailing was observed in both cases. Awareness should be paid to the amount of samples delivered from the spotting syringe because this step expresses the material concentration. A calibrated spotting device could be useful to avoid sample volume variation. However, the loaded volume area variation did not make any difference in the results. Sample solution stability. According to the experimental procedure described above, the preparation and loading of 20 samples on the plate takes approximately 30 min, which is enough time to allow for much material degradation, if it is degradable.

Therefore, it was necessary to investigate the solution stability during the analysis time. The results revealed that no degradation occurred during run time, and the solution was stable for at least 2 hours. The International Conference of Harmonization ICH guideline entitled stability testing of drug substances and products requires the stress testing to be carried out to elucidate the inherent stability characteristics of the active substance, and provide a rapid identification of differences that might result from changes in the manufacturing processes or the source sample Susceptibility to oxidation, hydrolytic, and photolytic stability are the required tests.

An ideal stability-indicating method is one that quantifies the standard drug alone and also resolves its degradation products. A relatively high concentration 1 mg of AMD was used to assure the detection of minor degradation products As described in the experimental section, different stress conditions were applied: boiling, acid, base hydrolysis, oxidation, and irradiation with UV light. From this investigation, it was clear that AMD was stable under all the stress conditions, as the chromatogram did not show any additional spots for degradation spots Fig.

Stress testing of AMD.

Lanes 1 and 8 are the AMD standard solution. Lanes from 2 to 6 are samples that have been subjected to alkali hydrolysis, oxidation, acid hydrolysis, ultraviolet irradiation, and boiling, respectively. Lane 7 is the capsule content subjected to moisture.

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The compatibility of AMD with the exipients used was also studied in the presence and absence of moisture. The stress testing results revealed that AMD was compatible with the combined exipients, whereas no more degradation products were observed when the stress testing experiments carried out on AMD-containing capsules Fig. It is evident from the results obtained previously that the proposed method gave satisfactory results with the analysis of AMD in bulk.

The label claim percentage was These results were compared with those obtained from the official method by statistical analysis with respect to the accuracy t-test and precision F-test. No significant differences were found between the calculated t- and F values were 1. This indicated similar accuracy and precision in the analysis of AMD in capsules form.

The present study represents the development and validation of a simple, accurate, and sensitive TLC method for quantitative determination of AMD. From an economics point of view, the method used the most simple and cost effective chromatographic technique; TLC. The method relies on the use of inexpensive equipment; scanner and software.

Additionally, all of the analytical reagents are inexpensive, and are available in any analytical laboratory. The proposed method could be recommended for routine use in quality control laboratories. National Center for Biotechnology Information , U. Int J Biomed Sci.

Hassan F. Askal , 1 Alaa S. Khedr , 1 Ibrahim A. Darwish , 1 and Ramadan M. Mahmoud 2. Alaa S. Ibrahim A. Ramadan M. Author information Article notes Copyright and License information Disclaimer. Corresponding Author: Ibrahim A. Box , Riyadh , Kingdom of Saudi Arabia.

Thin–layer Chromatography (TLC)

Received Feb 25; Accepted Mar Askal et al. Licensee Master Publishing Group. This article has been cited by other articles in PMC. Abstract A simple and accurate thin-layer chromatographic TLC method for quantitative determination of amantadine hydrochloride AMD was developed and validated. Keywords: amantadine hydrochloride, thin-layer chromatography, stability-indicating, pharmaceutical analysis.

INTRODUCTION

Open in a separate window. Figure 1. Preparation of standard solution An accurately weighed amount 1 g of AMD was transferred into a 10 ml calibrated flask and dissolved in about 5 ml of methanol. Preparation of capsule samples The contents of 20 capsules were mixed, and accurately weighed amount of the contents equivalent to mg of AMD was transferred into a 10 ml volumetric flask.

Thin-Layer Chromatography: Reagents and Detection Methods - PDF Drive

Preparation of detection reagent Forty grams of potassium iodide were as dissolved in ml of distilled water solution A. Methods and procedures Sample loading. Data processing and treatment The TLC chromatogram, as an image, is captured by the scanner and the image is then loaded into the Gel Works software. In the software, the following operations are performed: The series of spots to be manipulated are selected as a lane by lane creation function.

The background of the TLC chromatogram, if any, is subtracted. Figure 2. Method validation Linearity and limits of detection and quantitation. Figure 3. Nominated conc.