AUTHOR=Gervazoni Luiza F. O. , Gonçalves-Ozorio Gabriella , Ferreira-Paes Taiana , Silva Aline C. A. , Silveira Gabriel P. E. , Pereira Heliana M. , Pinto Douglas P. , Cunha-Junior Edézio F. , Almeida-Amaral Elmo E. 

TITLE=Analysis of 2′-hydroxyflavanone (2HF) in mouse whole blood by HPLC–MS/MS for the determination of pharmacokinetic parameters

JOURNAL=Frontiers in Chemistry

VOLUME=Volume 11 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/chemistry/articles/10.3389/fchem.2023.1016193

DOI=10.3389/fchem.2023.1016193

ISSN=2296-2646

ABSTRACT=Given the lack of investment and structure and the difficulty of metabolite isolation, promising natural product studies do not progress to preclinical studies, such as pharmacokinetics. 2’-Hydroxyflavanone (2HF) is a flavonoid that has shown promising results against different types of cancer and leishmaniasis. Herein, an HPLC–MS/MS method was developed and validated to accurately quantify the content of 2HF in BALB/c mouse blood. Chromatographic analysis was performed using a C18 column (5 μm, 150 × 4.6 mm). The mobile phase consisted of water (containing 0.1% formic acid), acetonitrile, and methanol (35/52/13 v/v/v) at a flow rate of 0.8 mL/min and total run time of 5.50 min with an injection volume of 20 µL. 2HF was detected by electrospray ionization in negative mode (ESI-) using MRM. The validated bioanalytical method showed satisfactory selectivity without significant interference with 2HF and the IS. In addition, good linearity (r=0.9969) was observed in the concentration range of 1 and 250 ng/mL. Moreover, the method showed satisfactory matrix effect results. The precision and accuracy intervals varied between 1.89 and 6.76% and 95.27 and 100.77%, respectively, fitting the approval criteria. No degradation of 2HF in the biological matrix was observed, since its stability after freeze–thaw cycles, short-term storage, postprocessing, and long-term storage showed deviations of less than 15%. Once validated, the method was successfully applied in an oral pharmacokinetic study with mouse blood, and the pharmacokinetic parameters of 2HF were determined. 2HF demonstrated a Cmax of 185.86 ng/mL, a Tmax of 5 min, and a half-life (t1/2) of 97.52 min.