Analytical Method Development, Validation and Optimization of Fluconazole Drug Using RP- HPLC

Aditi Chouksey; Nimita Manocha; Gurmeet Chhabra; Ritesh Patel; Gyanendra Singh Patel

doi:10.5281/zenodo.17553037

Research Paper | Open Access
Volume 02 | Issue 11 | Article Id IJSRT/

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Analytical Method Development, Validation and Optimization of Fluconazole Drug Using RP- HPLC
Aditi Chouksey* Nimita Manocha Gurmeet Chhabra Ritesh Patel Gyanendra Singh Patel
Department of Quality Assurance, Indore Institute of Pharmacy, Pithampur Road, opposite to IIM, Rau, Indore, Madhya Pradesh, Pin Code: 453331, India

Abstract

This study reported the development and validation of a simple, precise, and cost-effective Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method for the quantitative estimation of Fluconazole. Method development was systematically performed through nine optimization trials by varying the mobile phase composition, flow rate, and column type to achieve a sharp, symmetrical, and well-resolved peak with minimal tailing. The final optimized chromatographic conditions comprised a Kromasil C18 column (250 mm ? 4.6 mm, 5 ?m) with a mobile phase of Water: Acetonitrile (60:40 v/v) at a flow rate of 1.5 mL/min, which yielded a retention time of 2.231 minutes. The method exhibited excellent specificity, with no interfering peaks observed at the retention time of Fluconazole. Validation was performed in accordance with ICH guidelines, confirming the method?s reliability for routine quality control applications. System suitability parameters, including tailing factor, theoretical plates, and %RSD, were found to be within acceptable limits. The method demonstrated excellent linearity across 20?150% of the target concentration (r? = 0.999), along with high precision and accuracy. The assay of the drug substance was determined to be 99.49%, which was within the acceptable range. Overall, the developed RP-HPLC method proved to be simple, rapid, accurate, precise, robust, sensitive, and specific, making it suitable for the routine estimation of Fluconazole in pharmaceutical formulations.

Keywords

Fluconazole; RP-HPLC; Optimization; Validation; ICH guidelines; Analytical method; System suitability; Precision; Accuracy; Linearity; Specificity

Introduction

Pharmaceutical analysis plays a crucial role in ensuring the quality, safety, and efficacy of bulk drug substances and their formulations. Among the various analytical techniques available, High-Performance Liquid Chromatography (HPLC) has emerged as one of the most reliable and versatile tools for both qualitative and quantitative analysis in the pharmaceutical industry.[1,2] The data generated through HPLC can be either numerical, representing the exact amount of a compound present in a sample, or qualitative, confirming the presence or absence of specific analytes.[3,4] Owing to its high sensitivity, accuracy, and reproducibility, HPLC is employed at every stage of drug development from the analysis of raw materials to in-process controls and final product testing.[5,6] The specific objective of each analysis depends on the nature of the sample and the stage of pharmaceutical development. Therefore, a clear understanding of the fundamental principles of chromatography is essential to comprehend the operation and applications of HPLC in pharmaceutical analysis. In this research work, Fluconazole was selected as the model drug because it is a widely used triazole antifungal agent employed in the treatment of various systemic and superficial fungal infections. Despite its extensive clinical use, the available analytical methods for Fluconazole are often reported in combination with other drugs or involve complex mobile phase systems containing buffers or multiple solvents, which make them less suitable for routine analysis. Several analytical methods for the estimation of Fluconazole have been previously reported in the literature; however, most of these methods involve complex mobile phase compositions, buffer systems, or drug combinations, which limit their simplicity and routine applicability. Therefore, the present study aimed to develop a simple, reliable, and eco-friendly RP-HPLC method for the quantitative estimation of Fluconazole as a single drug component. Multiple optimization trials were performed using different ratios of Water and Acetonitrile as the mobile phase to achieve a sharp, symmetrical peak with an acceptable retention time and minimal tailing. The use of a this mobile phase not only reduced the organic solvent consumption but also made the method more environmentally friendly, cost-effective, and suitable for routine quality control analysis. Fluconazole, α-(2.4-diflurofenil)-α-(1H-triazol-1- methyl)-1H-1,2,4-triazol-1-ethano an antifungal medication, was discovered and developed by Pfizer. [7,8] Fluconazole is a first-generation triazole antifungal medication. It differs from earlier azole antifungals (such as ketoconazole) in that its structure contains a triazole ring instead of an imidazole ring. [9,10] The presence of a triazole and a difluoro phenyl produces potent antifungal activity but replacement of the usual imidazole group by triazole leads to improved selectivity.[11,12] Fluconazole is much less lipophilic than other azole antifungals and this leads to excellent penetration throughout the body, low protein-binding and water-solubility.[13,14] Fluconazole, was chosen for development on the basis of an optimal combination of antifungal efficacy, pharmacokinetic characteristics, aqueous solubility, and safety profile.[15]