• SABHYATHA T. S. Department of Pharmaceutical Chemistry, M. S. Ramaiah University of Applied Sciences, Bengaluru 560054, Karnataka, India
  • NARAYANA BABU Department of Pharmaceutical Chemistry, M. S. Ramaiah University of Applied Sciences, Bengaluru 560054, Karnataka, India



Method validation, Relative standard deviation, System suitability, Limit of detection, Limit of quantitation, Impurity-F, Gliclazide, RS-HPLC


Objective: The present study was undertaken with the objective of method validation of a rapid, simple, cost-effective HPLC method for the determination of related substances of Gliclazide.

Methods: A simple, rapid, and specific method for analysis of gliclazide by a sensitive high-performance liquid chromatographic method is described.

Validation of the method is carried out by USP and ICH guidelines. The method was validated for parameters like accuracy, precision, linearity, specificity, robustness, and system suitability. These proposed methods are suitable for the determination of title drugs in quality control laboratories in the pharmaceutical industries.

Results: The mobile phase used for the chromatographic runs consisted of (450 ml) of acetonitrile and (550 ml) of water. The separation was achieved on the LiChroCART Supersher RP-8 column, (250 mm × 4.0 mm, 5 µm), using isocratic mode. Drug peaks were well separated and were detected by a UV detector at 235 nm, the method was linear at the concentration range. Gliclazide limit of detection (LOD) and limit of quantification (LOQ) was 0.003 and 0.01 while LOD and LOQ for Impurity-F were 0.003 and 0.01 respectively.

Conclusion: The presented validated method is rapid, economic, simple, accurate, sensitive, robust, specific, and linear. It can be used for routine analysis of gliclazide.


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Stuart J. Practical HPLC method development by Lloyd R. Snyder (L. C. Resources, Inc.), Joseph J. Kirkland (Rockland Technologies, Inc.), and Joseph L. Glajch (DuPont Merck Pharmaceutical Co.). $84.95. J Am Chem Soc. 1998;120(14):3540.

Metzger S, Kolbesen BO. Application of HPLC for the analysis of organic additives in cleaning chemicals and cleaning mixtures. Solid State Phenom. 2005;103-104:221-6. doi: 10.4028/

Lal B, Kapoor D, Jaimini M. A review on analytical method validation and its regulatory perspectives. J Drug Delivery Ther. 2019;9(2):501-6. doi: 10.22270/jddt.v9i2.2403.

Ghosh S, Mandal S, Roy T, Ravikumar BVV. A Novel RP– HPLC analytical method development and validation of fulvestrant injection as per ICH guidelines. Egypt J Chem. 2021. doi: 10.21608/ejchem.2021.56503.3215.

Rouini MR, Mohajer A, Tahami MH. A simple and sensitive HPLC method for determination of gliclazide in human serum. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;785(2):383-6. doi: 10.1016/s1570-0232(02)00951-0, PMID 12554153.

Gosar A, Shaikh T, Joglekar A, Mahadik N. Development of analytical method for determination of p-toluenesulfonylurea and its impurities in gliclazide drug substances by using HPLC. J Pharmacol Res Dev. 2021;3(1):16-20. doi: 10.46610/JPRD.2021.v03i01.004.

Al Mahmud MA, Bhadra S, Haque A, Al Mamun ME, Haider SS. Development and validation of HPLC method for simultaneous determination of gliclazide and enalapril maleate in tablet dosage form. Dhaka Univ J Pharm Sci. 2015;13(1):51-6. doi: 10.3329/dujps.v13i1.21859.

Hossain MK, Sultan MZ, Rahman A, Uddin MG, Chowdhury AA, Rashid MA. RP-HPLC method development and validation for quantitative analysis of metformin and gliclazide in pharmaceutical formulations. J Sci Res. 2013;6(1):161-8. doi: 10.3329/jsr.v6i1.15963.

Nirupa G, Tripathi UM. RP-HPLC analytical method development and validation for simultaneous estimation of three drugs: glimepiride, pioglitazone, and metformin and its pharmaceutical dosage forms. J Chem. 2013;2013:1-8. doi: 10.1155/2013/726235.

Balaji N, Sultana S. Sensitive determination of related substances in pioglitazone hydrochloride by HPLC. Int J App Pharm. 2017;9(2):34. doi: 10.22159/ijap.2017v9i2.16828.

Rathinavel G, Uma Nath U, Valarmathy J, Samueljoshua L, Thanuja C, Ganesh M. RP-HPLC method for the simultaneous estimation of rosiglitazone and gliclazide in tablets. E-Journal of Chemistry. 2009;6(4):1188-92.



How to Cite

T. S., S., and N. BABU. “ANALYTICAL METHOD VALIDATION OF GLICLAZIDE RELATED SUBSTANCES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD”. International Journal of Current Pharmaceutical Research, vol. 14, no. 4, July 2022, pp. 36-41, doi:10.22159/ijcpr.2022v14i4.1999.



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