Flucytosine is fungicidal acting against Candida species but not against other common fungi. Flucytosine is structurally related to the cytostatic fluorouracil and to floxuridine. Flucytosine is chemically 6-amino-5-fluoro-1H-pyrimidin-2-one with molecular formula C4H4FN3O and molecular weight 129.09 g/mol. A new RP-HPLC method has been proposed for the quantification of Flucytosine in presence of internal standard Linezolid in pharmaceutical formulations and the method was validated as per ICH guidelines. Mobile phase consisting of a mixture of Tetra butyl ammonium hydrogen sulphate: Methanol (50:50, v/v) with a flow rate 1 mL/min and UV detection at 254 nm was used for the assay of Flucytosine in presence of internal standard Linezolid. Flucytosine was exposed to different stress conditions such as acidic, alkaline, oxidation and thermal degradation. Linearity was observed over the concentration range 1.0–100 µg/mL with linear regression equation y = 0.0238x - 0.0024 (R² = 0.9998). The LOQ was found to be 0.8937 µg/mL and the LOD was found to be 0.2938 µg/mL. The present method can be successfully applied for the pharmaceutical formulations, kinetics study and bioanalytical studies.
Cite this article:
Paladugu Venkata Naveen, Seru Ganapaty. A new stability indicating RP-HPLC method for the estimation of Flucytosine in presence of an internal standard. Research J. Pharm. and Tech. 2021; 14(1):289-294. doi: 10.5958/0974-360X.2021.00052.4
Paladugu Venkata Naveen, Seru Ganapaty. A new stability indicating RP-HPLC method for the estimation of Flucytosine in presence of an internal standard. Research J. Pharm. and Tech. 2021; 14(1):289-294. doi: 10.5958/0974-360X.2021.00052.4 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2021-14-1-52
1) Vermes A, Guchelaar HJ, Dankert J. Flucytosine: a review of its pharmacology, clinical indications, pharmacokinetics, toxicity and drug interactions. Journal of Antimicrobial Chemotherapy 46.2 (2000): 171–179.
2) Elias J. Anaissie, Michael R. McGinnis and Michael A. Pfaller, Clinical Mycology, 2nd Edition, 2009.
3) David N. Herndon, Total Burn Care, 3rd Edition, 2007.
4) Wade D and Sudlow G. Fluorometric measurement of 5-fluorocytosine in biological fluids. Journal of Pharmaceutical Sciences 62.5 (1973): 828–829.
5) Schiavone DJ, Page MD, Dawborn JK. Rapid microbiological assay for 5-fluorocytosine. Brazilian Medical Journal 17(4) (5889) (1973): 380–382.
6) Mathrusri Annapurna M, Rajasree B and Debi Prasad Pradhan. New Stability Indicating RP-UFLC Method for the Determination of Flucytosine – An Anti Fungal Agent. Acta Scientific Pharmaceutical Sciences. 3(5) (2019): 38-46.
7) Mohabbat Ullah, Md. Anisur Rahman. Development and validation of a Stability indicating RP-HPLC method for determination of Flucytosine and its process related impurities in injectable pharmaceuticals. International Journal of advances in pharmaceutical analysis. 5(1) (2018): 2277-9353.
8) Milind Ubale, Mahesh Shioorkar, Vilas Choudhari. A validated stability-indicating HPLC assay method for Flucytosine in bulk drug. International Journal of Innovations in Engineering and Technology. 8(1) (2017):155-161.
9) Ullrich Schwertschlag, Lisa M. Nakata and Joseph Gal. Improved procedure for determination of Flucytosine in human blood plasma by high-pressure liquid chromatography. Antimicrobial Agents and Chemotherapy 26.3 (1984): 303-305.
10) RW Bury, M L Mashford, H M Miles. Assay of Flucytosine (5-fluorocytosine) in human plasma by high-pressure liquid chromatography. Antimicrobial Agents and Chemotherapy 16.5 (1979): 529-532.
11) Serve KM, Yanez JA, Remsberg CM, Davies NM, Black ME. Development and validation of a rapid and sensitive HPLC method for the quantification of 5-fluorocytosine and its metabolites. Biomedical Chromatography 24.5 (2010): 556-561.
12) ICH Validation of analytical procedures: Text and methodology Q2 (R1), International Conference on Harmonization, 2005.
13) ICH Stability Testing of New Drug Substances and Products Q1A (R2), International Conference on Harmonization, 2003.