Simultaneous Determination and Method Validation of Fluconazole and its impurities by High Performance Thin Layer Chromatography Using Reflectance Scanning Densitometry
 
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School of Pharmacy, Suresh Gyan Vihar University, Jaipur, Rajasthan, India
CORRESPONDING AUTHOR
P. Sita Devi   

School of Pharmacy, Suresh Gyan Vihar University, Jaipur, Rajasthan, India
Publication date: 2017-10-24
 
Eurasian J Anal Chem 2013;8(1):39–49
 
KEYWORDS
ABSTRACT
A selective method based on HPTLC using reflectance scanning densitometry was developed and validated for the simultaneous separation and quantification of Fluconazole in formulations and its structurally related impurities. The chromatographic separation was accomplished on TLC aluminum plates precoated with silica gel 60 F254 as the stationary phase using a saturated mixture of butanol: water: acetic acid (8:2:1 v/v). For visualisation, the plate was dipped in to a modified anisaldehyde reagent and heated at 120°C for 30 minutes in a drying oven. Densitometric quantification was performed at 254 nm by reflectance scanning. Fluconazole appeared as a brown spot (Rf 0.67±0.02) and resolved well from the two impurities. The standard fluconazole curve is linear (r =>0.9995) over a concentration range of 100-800ng/spot. Recovery from tablet formulation was statistically equal to 100% .The limits of detection and quantification were 91.39 and 304.66 ng/spot respectively. The precision of the method with respect to concentration is acceptable with a relative standard deviation of 0.73%. The proposed method is specific for fluconazole in the presence of its structurally related impurities and proved to be a valuable complimentary method for impurity profiling and quality control.
 
REFERENCES (20)
1.
Morrow JD (1991) Fluconazole: a new triazole antifungal agent. Am J Med Sci, 302:129.
 
2.
Pasko MT, Piscitelli SC, and Van Slooten AD (1990) Fluconazole: a new triazole antifungal agent DICP, The Annals of Pharmacotherapy: Vol. 24:860.
 
3.
Oliver RJ, Dhaliwal HS, Theaker ED, Pemberton MN (2004) Patterns of antifungal prescribing in general dental practice - British Dental Journal, 196:701.
 
4.
Ellis ME, Clink H, Ernst P, Halim MA, Padmos A, Spence D, Kalin M, Hussain Qadri SM, Burnie J and Greer W (2005) Controlled study of fluconazole in the prevention of fungal infections in neutropenic patients with haematological malignancies and bone marrow transplant recipients European Journal of Clinical Microbiology & Infectious Diseases, 13:11.
 
5.
Presterl E, Graninger W (1994) Efficacy and safety of fluconazole in the treatment of systemic fungal infections in pediatric patients European Journal of Clinical Microbiology & Infectious Diseases, 13:347.
 
6.
Goodman JL, Winston DJ, Greenfield RA (1992), A controlled trial of fluconazole to prevent fungal infections in patients undergoing bone marrow transplantation - New England Journal of Medicine, 326: 845.
 
7.
C Fasano, JO'Keeffe, D Gibbs (1994), Fluconazole treatment of neonates and infants with severe fungal infections not treatable with conventional agents- European Journal of Clinical Microbiology & Infectious, 13: 351.
 
8.
Lamb DC, Kelly DE, White TC, and Kelly SL (2000) The R467K Amino Acid Substitution in Candida albicans Sterol 14α- Demethylase Causes Drug Resistance through Reduced Affinity Antimicrob Agents Chemother, 44: 63.
 
9.
Ballard SA, Ellis SW and Troke PF (1990), Interaction of microsomal cytochrome P-450 isolated from, 4spergillus fumigatus with fluconazole and voriconazole Journal of Medical and Veterinary Mycology, 28: 327.
 
10.
Pasko MT, Piscitelli SC, Van Slooten AD (1990), Fluconazole: a new triazole antifungal agent, DICP, 24: 860.
 
11.
Vanden Bossche H (1985) Biochemical targets for antifungal azole derivatives: hypothesis on the mode of action. Curr Top Med Mycol. 1:313-51.
 
12.
International Conference on Harmonization. ICH Harmonized Tripartite Guidelines. Q3A(R): Impurities in New Drug Substances. Fed. Reg. 2003, 68 (68), 6924-6925.
 
13.
International Conference on Harmonization. ICH Harmonized Tripartite Guidelines Q3B(R): Impurities in New Drug substances. Fed. Reg. 2003, 68 (220), 64628- 64629.
 
14.
Inagaki K, Takagi J, Lor E, Okamoto MP, Gill MA (1992) Determination of fluconazole in human serum by solid-phase extraction and reversed-phase highperformance liquid chromatography. Ther. Drug Monit.14: 306.
 
15.
Wood P, and Tarbi M (1986) Gas chromatography method for the determination of fluconazole, a noval antifungal agent, in human plasma and urine. J. Chromatogr. 383:179.
 
16.
Debruyne D, Ryckelynck JP, Bigot MC, Moulin M (1988) Determination of fluconazole in biological fluids by capillary column gas chromatography with a nitrogen detector. J. Pharmaceut. Sci. 77: 534.
 
17.
Hosotsubo, K.K.; Hosotsubo, H.; Nishijima, M.K.; Okada, T.; Taenaka, N.; Yoshiya, I (1990). Rapid determination of serum levels of a new antifungal agent, fluconazole, by high-performance liquid chromatography. J. Chromatogr. A, 529:223.
 
18.
Renger (2001) B. Planar chromatographic procedures in pharmaceutical analysis. J. AOAC Int. 84:1217.
 
19.
ICH, Text on Validation of Analytical Procedure – ICH Harmonized Tripartite Guideline, 1994.
 
20.
ICH, Validation of analytical procedures: Methodology- ICH Harmonized Tripartite Guideline, 1996.
 
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