HPLC-BASED BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF POSACONAZOLE IN SPIKED RAT PLASMA

Authors

  • INDUJA GOVINDAN Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, India
  • ANJANA A. KAILAS Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, India https://orcid.org/0009-0005-5796-792X
  • ABUTWAIBE KA Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, India https://orcid.org/0000-0002-8395-3506
  • THAMIZHARASAN ANNADURAI Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, India https://orcid.org/0009-0006-4470-8061
  • ANUP NAHA Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal-576104, India

DOI:

https://doi.org/10.22159/ijap.2025v17i5.54731

Keywords:

Posaconazole, Bioanalytical method, HPLC, Green chemistry, Stability

Abstract

Objective: Posaconazole (PSC), a highly effective broad-spectrum triazole antifungal, is widely used to treat invasive fungal infections. Accurate and consistent quantification of PSC in biological matrices, such as plasma, is vital for supporting pharmacokinetic and therapeutic drug monitoring studies. To develop and validate a sensitive, robust, and sustainable bioanalytical HPLC method for the quantification of posaconazole in plasma.

Methods: The method utilises a simple protein precipitation technique for sample preparation, avoiding more hazardous or solvent-intensive procedures. Chromatographic separation was achieved using an isocratic elution with a mobile phase composed of acetonitrile and water acidified with 0.5% acetic acid (44:56), delivered at a flow rate of 0.7 ml/min. Unlike most reported methods that use phosphate buffers, this method employs water acidified with 0.5% v/v of acetic acid, enhances environmental compatibility and avoids buffer salt disposal and system fouling issues. Detection was performed at 262 nm using a Shimadzu LC-2010CHT system with a dual-wavelength UV detector and a photodiode array (SPD-M20A, Prominence series).

Results: The method exhibited excellent linearity over the 0.25–32 µg/ml range (r² = 0.9999). Intra and inter-day precision and accuracy complied with ICH M10 bioanalytical validation guidelines, with %RSD values below 2% and recoveries ranging from 97.7±0.07 to 101.12±0.03%. Stability under all relevant conditions was confirmed. Environmental sustainability was evaluated using AGREE and GAPI tools, an approach not previously applied to posaconazole bioanalysis, highlighting its novelty and sustainable design for routine use.

Conclusion: A validated bioanalytical HPLC method for posaconazole quantification in plasma was successfully developed, is suitable for routine pharmacokinetic and therapeutic drug monitoring applications.

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Published

07-09-2025

How to Cite

GOVINDAN, I., KAILAS, A. A., KA, A., ANNADURAI, T., & NAHA, A. (2025). HPLC-BASED BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF POSACONAZOLE IN SPIKED RAT PLASMA. International Journal of Applied Pharmaceutics, 17(5), 471–480. https://doi.org/10.22159/ijap.2025v17i5.54731

Issue

Vol 17, Issue 5 (Sep-Oct), 2025

Section

Original Article(s)

Copyright (c) 2025 INDUJA GOVINDAN, ANJANA A. KAILAS, ABUTWAIBE KA, THAMIZHARASAN ANNADURAI, ANUP NAHA

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This work is licensed under a Creative Commons Attribution 4.0 International License.

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