Validation of high-performance liquid chromatographic method for analysis of fluconazole in microemulsions and liquid crystals

Hilris Rocha E Silva,Maria Palmira Daflon Gremião,Gabriela Marielli Da Luz,Fernanda Kolenyak Dos Santos,Marlus Chorilli

doi:10.1590/s1984-82502014000200018

Hilris Rocha E Silva, Maria Palmira Daflon Gremião + Show 3 more

Open Access

https://doi.org/10.1590/s1984-82502014000200018

Copy DOI

Abstract

In recent decades, there has been a significant increase in the incidence of fungal diseases. Certain fungal diseases cause cutaneous lesions and in the usual treatment, generally administred orally, the drug reaches the site of action with difficulty and its concentration is too low. An approach much explored in recent years is the development of nanotechnology-based drug delivery systems, and microemulsions (ME) and liquid crystals (LC) are promising. ME and LC were developed with oleic acid or copaiba oil as the oil phase, propoxyl (5OP) ethoxyl (20 OE) cetyl alcohol as surfactant and water. An analytical method to assess the incorporation of fluconazole (FLU) in the systems under study was validated according to guidelines of the International Conference on Harmonization (ICH) guidelines and the Brazilian Food, Drug and Sanitation Agency (ANVISA). The method was conducted on a C18-RP column (250 × 4.6 mm i.d.), maintained at room temperature. The mobile phase consisted of acetonitrile and water (50:50, v/v), run at a flow rate of 1.0mL/min and using ultraviolet detection at 210nm. The chromatographic separation was obtained with a retention time of 6.3min, and was linear in the range of 20-400 µg/mL (r2=0.9999). The specificity showed no interference of the excipients. The accuracy was 100.76%. The limits of detection and quantitation were 0.057 and 0.172 µg.mL-1, respectively. Moreover, method validation demonstrated satisfactory results for precision and robustness. The proposed method was applied for the analysis of the incorporation of FLU in ME and LC, contributing to improve the quality control and to assure the therapeutic efficacy.

Highlights

Fungal infections are considered a major problem for public health
In order to investigate the ME and liquid crystals (LC) regions, a ternary phases diagram was constructed, containing as oil phase copaiba oil obtained of cooperatives of Acre (Brazil) (CO), copaiba oil purchased from Sigma (OS) or oleic acid (OA)
Accuracy To confirm the accuracy of the proposed method, a sample of fluconazole working solution was prepared in triplicate at three different concentrations, 20, 40 and 200 μg.mL-1, within the analytical curve

Summary

Introduction

Fungal infections are considered a major problem for public health. A significant group of such infections are the dermatomycoses, involving the skin, especially the stratum corneum, and appendages such as nails, hair and mucous membranes (oral and vaginal) (Negroni, 2010; Erchiga, Fiorencio, 2005; Bachhav, Patrale, 2009).Fluconazole – FLU (Figure 1) is a white, slightly soluble in water, soluble in ethanol, methanol and acetone and very slightly soluble in toluene. Fungal infections are considered a major problem for public health. A significant group of such infections are the dermatomycoses, involving the skin, especially the stratum corneum, and appendages such as nails, hair and mucous membranes (oral and vaginal) (Negroni, 2010; Erchiga, Fiorencio, 2005; Bachhav, Patrale, 2009). Like that of the other azoles, is related to its ability to alter the membrane permeability of yeasts and other fungi by inhibiting the synthesis of ergosterol. Inhibition of P-450 lanosterol 14-alpha-demethylase dependent causes accumulation of sterols methylated depletion ergosterol and inhibition of cell growth (Alnaim et al, 2007; Jovanović et al, 2005; Pore et al, 2006)

Objectives

Methods

Results