HPLC-DAD for the determination of three different classes of antifungals: method characterization, statistical approach, and application to a permeation study.
This study describes and characterizes methods for high-performance liquid chromatography diode array detection (HPLC-DAD) analysis of formulations containing molecules with antifungal activity of three different classes: terbinafine and butenafine (allylamines), miconazole and fluconazole (azoles), and geraniol, neral and geranial (monoterpenes). All methods used the same chromatographic column (RP18 ), enabling the analysis to be performed in a single batch. The specificity was extensively discussed through the establishment of purity peak methods. The analytical parameters (linearity, precision and accuracy) were calculated and discussed in detail using specific statistical approaches. All substances showed satisfactory results for chromatographic and analytical parameters. Limits of 1.3% to mean repeatability and 2.0% for intermediate precision are suggested as acceptance criteria in validation of methods by HPLC-DAD, in situations where there is no extensive pretreatment of the samples. The methods proved to be robust and significant factors were discussed regarding their influence on chromatographic parameters (retention time, resolution, tailing factor and column efficiency). Finally, the application of the developed methods was demonstrated by the results of a permeation study of the antifungal agents through bovine hoof membranes.
Pharmaceutical Quality Control Laboratory, School of Pharmacy, Federal University of Rio Grande do Sul, 90610-000, Porto Alegre, Brazil.
Diogo
D
Miron
Alini
A
Lange
Aline R
AR
Zimmer
Paulo
P
Mayorga
Elfrides E S
EE
Schapoval
D000818:Animals; D000935:Antifungal Agents / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006724:Hoof and Claw / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D010539:Permeability; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000935:Antifungal Agents
butenafine; citral; fluconazole; geraniol; miconazole; terbinafine