Fluorescence polarization immunoassay of colchicine

Elena A Zvereva; Anatoly V Zherdev; Andey A Formanovsky; Ramadan A Abuknesha; Sergei A Eremin; Boris B Dzantiev

doi:10.1016/j.jpba.2018.07.008

Fluorescence polarization immunoassay of colchicine

J Pharm Biomed Anal. 2018 Sep 10:159:326-330. doi: 10.1016/j.jpba.2018.07.008. Epub 2018 Jul 11.

Authors

Elena A Zvereva¹, Anatoly V Zherdev¹, Andey A Formanovsky², Ramadan A Abuknesha³, Sergei A Eremin⁴, Boris B Dzantiev⁵

Affiliations

¹ A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, 119071, Russia.
² M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, 117997, Russia.
³ King's College London, Strand, London, WC2R 2LS, United Kingdom.
⁴ A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, 119071, Russia; Department of Chemistry, M.V. Lomonosov Moscow State University, Moscow, 119991, Russia.
⁵ A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, 119071, Russia. Electronic address: dzantiev@inbi.ras.ru.

PMID: 30025297
DOI: 10.1016/j.jpba.2018.07.008

Abstract

In this study, a fluorescence polarization immunoassay (FPIA) technique was developed to determine colchicine (COL), an alkaloid of noxious plants of the order Liliales that is used in a number of medications to treat gout. An optimal combination of the polyclonal antibody and the antigen labelled with fluorescein isothiocyanate (FITC) was selected. Conditions for the competitive interaction of the antigen in the tested samples and its fluorophore conjugate (COL-FITC) with anti-COL antibodies were optimised, and the analytical characteristics of the assay were determined. The developed FPIA was characterised by a detection limit of 1.8 ng/mL and a detectable analyte concentration range of 4.1-74.3 ng/mL. The duration of the analysis was 10 min. The applicability of the developed FPIA for quality control of ready-made drug formulations and for the estimation of COL content in various matrices (urine, milk), with recovery values ranging from 79 to 108%, was demonstrated.

Keywords: Colchicine; Fluorescence polarization immunoassay.

MeSH terms

Animals
Chemistry, Pharmaceutical / methods*
Colchicine / analysis*
Colchicine / metabolism*
Fluorescence Polarization Immunoassay / methods
Gout Suppressants / analysis*
Gout Suppressants / metabolism*
Humans
Milk / chemistry
Milk / metabolism

Substances

Gout Suppressants
Colchicine