Quantification of dapaconazole in human plasma using high-performance liquid chromatography coupled to tandem mass spectrometry: application to a phase I study.
A simple, selective and sensitive method based on high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) has been developed for the determination of dapaconazole in human plasma using tioconazole as internal standard. The drugs were extracted from plasma by liquid-liquid extraction with ether/hexane (80/20, v/v). The chromatography separation was performed on a Genesis(®) C18 reversed phase analytical column 4μm (100×2.1mm i.d.) with a mobile phase of methanol/acetonitrile/water (80/10/10, v/v/v)+ammonium acetate (0.5mM). Dapaconazole was quantified using a mass spectrometer with an electrospray source in the ESI positive mode (ES+) configured for multiple reaction monitoring (MRM) to monitor the transitions 415.1>159.2 and 387.0>131.0 for dapaconazole and tioconazole, respectively. The method had a chromatography run time of 3.8min and a linear calibration curve over the range 0.2-100ng/mL (r=0.9998). The lower limit of quantification (LLOQ) was 0.2ng/mL. The precision and accuracy values of the assay were within ±10%. The stability tests indicate no significant degradation under the conditions of the experiment. This method was used for a phase I study of topical administration of dapaconazole tosylate in healthy human male volunteers.
Galeno Research Unit, Latino Coelho St., 1301, Parquel Taquaral, 13087-010 Campinas, SP, Brazil; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil.
Fernanda C
FC
de Moraes
Galeno Research Unit, Latino Coelho St., 1301, Parquel Taquaral, 13087-010 Campinas, SP, Brazil; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil.
Samara F
SF
Bittencourt
Department of Pharmaceutical and Toxicological Chemistry, Federico II University, Naples, Italy.
Elisa
E
Perissutti
Department of Pharmaceutical and Toxicological Chemistry, Federico II University, Naples, Italy.
Francesco
F
Frencentese
Galeno Research Unit, Latino Coelho St., 1301, Parquel Taquaral, 13087-010 Campinas, SP, Brazil.
André M M
AM
Arruda
Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil.
Lu Shi
LS
Chen
Galeno Research Unit, Latino Coelho St., 1301, Parquel Taquaral, 13087-010 Campinas, SP, Brazil.
Tainah
T
Babadópulos
Galeno Research Unit, Latino Coelho St., 1301, Parquel Taquaral, 13087-010 Campinas, SP, Brazil; Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil; Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil. Electronic address: denucci@gilbertodenucci.com.
Gilberto
G
De Nucci
D000935:Antifungal Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D007093:Imidazoles / Q000097:blood*; D057230:Limit of Detection; D008297:Male; D053719:Tandem Mass Spectrometry / Q000379:methods
C000593721:1-(2-(2,4-dichlorophenyl)-2-(4-(trifluoromethyl)benzyloxy)ethyl)-1H-imidazole; D000935:Antifungal Agents; D007093:Imidazoles
Antifungal; Blood; HPLC; Healthy volunteers; Imidazole; MS/MS