TY - JOUR

T1 - Development and validation of ultrafast LC–MS/MS method for quantification of anti-influenza agent camphecene in whole rat blood using dried blood spots and its application to pharmacokinetic studies

AU - Rogachev, Artem D.

AU - Yarovaya, Olga I.

AU - Ankov, Sergey V.

AU - Khvostov, Mikhail V.

AU - Tolstikova, Tatyana G.

AU - Pokrovsky, Andrey G.

AU - Salakhutdinov, Nariman F.

N1 - Funding Information: This study is supported by Russian scientific foundation grant (No. 15-13-00017 ). Publisher Copyright: © 2016 Elsevier B.V. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.

PY - 2016/11/15

Y1 - 2016/11/15

N2 - A fast, selective and sensitive procedure for quantitation of the camphor-based anti-influenza agent camphecene in whole rat blood was developed and validated using dried blood spots and LC–MS/MS. The method was validated according to recommendations of the FDA and EMA in terms of selectivity, linearity, accuracy, precision, recovery, matrix factor, stability, and carry-over. Sample preparation included spotting 20 μL of whole blood taken from the tail vein onto the paper, drying and extracting the analyte, followed by evaporation of the solvent and analysis of the residue. HPLC separations were run on a reversed-phase microcolumn; the time of analysis was less than 2 min. MS/MS detection was performed on a triple quadrupole mass-spectrometer using multiple reaction monitoring (MRM) mode. Transitions 196.4 → 122.2/153.3 and 152.2 → 93.1/107.2 were monitored for camphecene and 2-adamantylamine hydrochloride (internal standard), respectively. The intra- and inter-day precisions and accuracies, matrix factor, carry-over and recovery were within acceptable limits. Despite low extraction recovery (less than 2%), the sensitivity of the method was enough to detect the analyte in the concentration range 50–2500 ng/mL. The application of the method was shown in pharmacokinetic studies of camphecene in rats at a dose of 10 mg/kg.

AB - A fast, selective and sensitive procedure for quantitation of the camphor-based anti-influenza agent camphecene in whole rat blood was developed and validated using dried blood spots and LC–MS/MS. The method was validated according to recommendations of the FDA and EMA in terms of selectivity, linearity, accuracy, precision, recovery, matrix factor, stability, and carry-over. Sample preparation included spotting 20 μL of whole blood taken from the tail vein onto the paper, drying and extracting the analyte, followed by evaporation of the solvent and analysis of the residue. HPLC separations were run on a reversed-phase microcolumn; the time of analysis was less than 2 min. MS/MS detection was performed on a triple quadrupole mass-spectrometer using multiple reaction monitoring (MRM) mode. Transitions 196.4 → 122.2/153.3 and 152.2 → 93.1/107.2 were monitored for camphecene and 2-adamantylamine hydrochloride (internal standard), respectively. The intra- and inter-day precisions and accuracies, matrix factor, carry-over and recovery were within acceptable limits. Despite low extraction recovery (less than 2%), the sensitivity of the method was enough to detect the analyte in the concentration range 50–2500 ng/mL. The application of the method was shown in pharmacokinetic studies of camphecene in rats at a dose of 10 mg/kg.

KW - Anti-influenza

KW - Camphecene

KW - Dried blood spots

KW - LC–MS/MS

UR - http://www.scopus.com/inward/record.url?scp=84991584210&partnerID=8YFLogxK

U2 - 10.1016/j.jchromb.2016.10.009

DO - 10.1016/j.jchromb.2016.10.009

M3 - Article

C2 - 27750194

AN - SCOPUS:84991584210

VL - 1036-1037

SP - 136

EP - 141

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

ER -