|
ИСТИНА |
Войти в систему Регистрация |
ИСТИНА ИНХС РАН |
||
Design of a sensitive fluorescent polarization immunoassay for rapid screening of milk for cephalexinстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 12 июля 2019 г.
- Авторы: Beloglazova Natalia V., Eremin Sergei A.
- Журнал: Analytical and Bioanalytical Chemistry
- Том: 407
- Номер: 28
- Год издания: 2015
- Издательство: Springer Verlag
- Местоположение издательства: Germany
- Первая страница: 8525
- Последняя страница: 8532
- DOI: 10.1007/s00216-015-9006-6
- Аннотация: In this paper we describe the development of a sensitive, fast, and easily performed fluorescence polarization immunoassay for determination of cephalexin in milk. The experimental work was performed to increase sensitivity and specificity. Therefore, the structures of the tracers were varied by synthesis of both cephalexin (CEX) and cephalotin (CET) conjugates with a variety of fluorescent labels. Two rabbit antisera containing antibodies against cephalexin and cephalotin were tested in homologous and heterologous combinations with the tracers. For every working antibody–tracer combination, the analytical conditions and cross-reactivity for structural analogues—cephalosporins and other antibiotics that could also be present in milk—were determined. It was found that the highest sensitivity was achieved by use of the homologous pair CET–EDF–anti-CET antibody (limit of detection (LOD) 0.4 μg kg−1 for standard solutions prepared in buffer), but this combination was not appropriate because of high cross-reactivity with CET. For subsequent experiments, therefore, CEX– EDF–anti-CEX antibody were chosen (LOD 0.8 μg kg−1 for standard solutions prepared in buffer). Part of this manuscript is devoted to the variation of precipitation agents for pretreatment of milk before analysis; milk is an extremely complicated matrix. The optimum protein precipitation agent was methanol. This technique for cephalexin determination was characterized by a limit of detection of 1 μg kg−1. The method was validated by using naturally contaminated and spiked milk samples. The results obtained corresponded very well with those obtained by HPLC, which was used as confirmation method.
- Добавил в систему: Еремин Сергей Александрович