Development and Validation of High Performance Planar Chromatography for Estimation of Gentamicin by Fluorophore Generation

Author: Arka Bhattacharjee, Arpan Chakraborty, Baishakhi Mondal, Nitai Chand Chaulya, Goutam Mukhopadhyay and Alpana Majumder

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Abstract

This study presents the development and validation of an HPTLC method for estimating Gentamicin in bulk powder, pharmaceutical dosage forms, and blood plasma samples. The chromatography separation was conducted on pre-coated aluminum plates using silica gel 60 F254 as the stationary phase and a mobile phase comprising chloroform, methanol, and 25% ammonia solution. The developed method was able to successfully separate and detect the various components of Gentamicin, despite the challenges of optimizing the separation and detection parameters. The method produced compact and well-resolved bands, which is important for accurate analysis. By demonstrating excellent linearity (r2 = 0.9996) in the calibration plots, the method enables the quantification of Gentamicin within the concentration range of 60-200 ng/band at 436 nm. Furthermore, the method's precision, accuracy, robustness, specificity, limit of detection, and limit of quantification were validated according to the International Conference on Harmonization (ICH) guidelines. The results of the study confirm that the developed method is suitable for routine quality control testing of Gentamicin in bulk drug, marketed formulations, and plasma samples. This means that the method can be used to reliably ensure the quality of Gentamicin and guide its therapeutic use.

Keywords

Gentamicin, Planar-chromatography, Derivatization, HPTLC, Flurophore, Optimization

Conclusion

The developed HPTLC technique enables the quantitative analysis of GMS in bulk drug, marketed formulation, and plasma samples. With RSD values of 2%, the method demonstrates satisfactory accuracy. The recovery percentages for GMS (100.925–101.795%) further confirm the efficiency and reliability of the method. In addition, the fingerprint profiling of chromatograms obtained from GMS can be used for comparing and evaluating commercial samples. Compared to HPLC, HPTLC offers advantages such as shorter processing times, lower sample requirements, optimized extractions using cost-effective chemicals, and smaller mobile phase volumes. This quick, easy, and sensitive HPTLC procedure serves as a valuable quality control tool for assessing the aerial GMS in different test samples.

References

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How to cite this article

Arka Bhattacharjee, Arpan Chakraborty, Baishakhi Mondal, Nitai Chand Chaulya, Goutam Mukhopadhyay and Alpana Majumder (2023). Development and Validation of High Performance Planar Chromatography for Estimation of Gentamicin by Fluorophore Generation. Biological Forum – An International Journal, 15(5): 1543-1549.