Development and Validation of HPLC Method for Efinaconazole: Application to Human Nail Permeation Studies

Efinaconazole is the first azole derivative approved by FDA for the topical treatment of onychomycosis. The objective of present study was to develop and validate HPLC method for estimation of efinaconazole in ex vivo human nail permeation study samples. The chromatographic analysis was performed on a HPLC system equipped with diode array detector. The efinaconazole and internal standard (IS) were extracted from the human nail samples by using the protein precipitation method. The samples were injected on to 5 mu m Polar C-18 100 angstrom, 4.6 mm x 150 mm column. The mobile phase consisted of 0.01 M potassium dihydrogen phosphate: acetonitrile (36:64) and eluent was monitored at 205 nm. The chromatographic separation of drug and analyte was achieved using isocratic elution at flow rate of 1 mL/min with a total run time of 15 min. The efinaconazole and IS were eluted at 6.4 +/- 0.5 and 8.3 +/- 0.5 min, respectively. The developed method was validated as per FDA guidelines, and the results met with acceptance criteria. The method developed was specific, and the analyte concentrations were linear at range of 50 to 10000 ng/mL (R-2 >= 0.9981). The validated HPLC method was applied for quantifying efinaconazole in human nail permeation study samples. The permeation of efinaconazole was increased by twofolds with Labarfac CC (15135.4 +/- 2233.9 ng/cm(2)) compared to formulations containing Transcutol P (6892.0 +/- 557.6 ng/cm(2)) and Labrasol (7266.1 +/- 790.6 ng/cm(2)). The study results demonstrate that developed efinaconazole HPLC method can be employed for formulation evaluation and clinical studies.

Automated summary

A validated HPLC method for quantifying efinaconazole in ex vivo human nail permeation study samples was developed and applied. The method showed good specificity and linearity in the concentration range of 50 to 10000 ng/mL. The study also demonstrated that Labarfac CC significantly increased the permeation of efinaconazole compared to other formulations.