Quantitative determination and pharmacokinetic study of fusaricidin A in mice plasma and tissues using ultra-high performance liquid chromatography-tandem mass spectrometry

Authors: HARON, MONA - University Of Mississippi; AVULA, BHARATHI - University Of Mississippi; QUI, SHI - University Of Mississippi; LI, XING-CONG - University Of Mississippi; ASHFAQ, MOHAMMAD - University Of Mississippi; BAE, JI-YEONG - University Of Mississippi; GUAN, SHAOHUA - Agricen Sciences; HINCHEE, MAUD - Agricen Sciences; KHAN, IKHLAS - University Of Mississippi; KHAN, SHABANA - University Of Mississippi

Submitted to: Journal of Pharmaceutical and Biomedical Analysis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/18/2019
Publication Date: 3/19/2019
Publication URL: https://handle.nal.usda.gov/10113/6471201
Citation: Haron, M.H., Avula, B., Qui, S., Li, X., Ashfaq, M.K., Bae, J., Guan, S., Hinchee, M., Khan, I.A., Khan, S.I. 2019. Quantitative determination and pharmacokinetic study of fusaricidin A in mice plasma and tissues using ultra-high performance liquid chromatography-tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis. 170:187-192. https://doi.org/10.1016/j.jpba.2019.03.042.
DOI: https://doi.org/10.1016/j.jpba.2019.03.042

Interpretive Summary: There has been an increasing demand for new, safe and effective antibacterial and antifungal agents, due to the emerging of antibiotic resistant bacteria and opportunistic fungal infections. Recently a group of naturally occurring cyclic peptides, “Fusaricidins or LI-F” has emerged as promising pharmacologically active compounds for the development of antibacterial and antifungal agents. Fusaricidins demonstrated strong in vitro antibacterial activity against gram-positive bacteria such as Staphylococcus aureus, Micrococcus luteus, and Bacillus subtilis as well as fungal pathogens e.g. Candida spp. and Cryptococcus neoformans. Since there is no report of a quantitative analytical method for FA in biological matrices, a highly sensitive liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the quantitative analysis of FA. UHPLC-MS/MS is considered as the method of choice for quantitative analysis of the analyte in the biological matrices due to its high sensitivity and selectivity. The analysis time is rapid and involves simple sample pre-treatment which is less complicated. The developed method was successfully applied to study the pharmacokinetics and tissue distribution of FA in CD1 mice after intravenous (I.V.) administration. The data generated will be helpful in guiding future in vivo efficacy studies of this promising class of antimicrobial compounds for potential pharmaceutical development.

Technical Abstract: Fusaricidins are a family of cyclic lipodepsipeptides that convey antifungal and antibacterial activity. Fusaricidin A is one of the Fusaricidins major compounds and it is showing promising activity against fungi and bacteria. In the present study, a fast and sensitive ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the analysis of fusaricidin A in mice plasma, liver, kidney and brain tissues. The instrument was operated in positive electrospray ionization mode. Multiple reaction monitoring (MRM) mode was performed with ion pairs of m/z: 883.5 to 256.3, 883.5 to 197.2 and 883.5 to 72.1 for fusaricidin A. The method was validated for linearity, repeatability, accuracy, stability, limits of detection (LOD) and limits of quantification (LOQ). The LOD and LOQ were 0.01 and 0.05 ng/mL for plasma and tissues, respectively. The calibration of 10 - 200 ng/mL was linear (r2 = 0.99). Precision and accuracy values were found to be < 10 % (within acceptable limit). The pharmacokinetic and tissue distribution characteristics of FA were determined in plasma, liver, kidney and brain of CD1 mice after I.V. administration of a single dose of 15 mg/kg body weight. Highest plasma concentration (Cmax) was calculated to be 4169.97 ± 50 ng/mL with a tmax of 0.08 h. The plasma clearance rate of fusaricidin A was 397.6 ± 203 mL/h with a t1/2 of 2.2 ± 0.5 h and apparent volume of distribution during the terminal phase (Vz) of 979.2 ± 318 mL. The highest tissue concentration (Cmax) was found in the liver (219 ± 14 ng/mg) at a tmax of 0.08 h followed by the kidneys (38.6 ± 16 ng/mg) at tmax of 0.2 h. Fusaricidin A was poorly distributed to the brain with a Cmax of 0.45 ± 0.2 ng/mg and a tmax of 0.08 h. The method for quantitative analysis and pharmacokinetic data provided will support the development of various formulation approaches and therapeutic application for future clinical studies.