Determination of the veterinary drug maduramicin in food by fluorescence polarisation immunoassay

Abstract

A fluorescence polarisation immunoassay (FPIA) using a specific polyclonal antiserum for the detection of maduramicin (MD) was developed and optimised. The polyclonal antiserum was produced against MD linked to bovine serum albumin. Fluorescein-labelled MD (tracer) was synthesised by N-hydroxysuccinimide active ester method and purified using thin layer chromatography. The developed FPIA for MD had a dynamic range from 0.01 to 5.6 lg mL )1 with an IC 50 value of 0.16 lg mL )1 and a limit of detection of 0.002 lg mL )1 . Recoveries from chicken muscle, fat and egg samples spiked at 0.25, 5 and 10 lg g )1 levels were 82-130%. The FPIA results from analysis of incurred residues in chicken muscle samples showed that the simple procedure is viable.

Development of an indirect competitive ELISA based on immunomagnetic beads’ clean-up for detection of maduramicin in three chicken tissues

“…Our developed method was challenged with authentic samples collected from local markets in Beijing, China. For the sample pre-treatment, methanol was selected as extracting agent to extract MD from chicken tissues, due to its high extraction efficiency of MD with less influence on immunoassay (Wang et al, 2008). The extracts were 1-10-fold diluted with 0.01 mol/L PBS (pH 7.4) for muscle, skin and fat, and 1-15-fold diluted for liver before IMBs-based clean-up.…”

“…Furthermore, MD has been widely used as a growth promoter to improve feed conversion rate in ruminants (Le-Minh, Khan, Drewes, & Stuetz, 2010). However, the extensive or illegal use of MD can lead to intoxication of animals and residues in chicken tissues (Wang, Zhang, Murtazina, Eremin, & Shen, 2008), which constitutes a potential hazard for both human and environment's health (Sharma, Bhalla, Varma, Jain, & Singh, 2005). For example, MD could increase coronary blood flow and induce coronary dilation (Pereira et al, 2016), and made the situation even worse on the patients with coronary-artery-related diseases (Elliott, Kennedy, & McCaughey, 1998).…”

“…Most of them are based on instrumental analysis such as high-performance liquid chromatography (HPLC) with either UV or fluorescent detector and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) (De Jong et al, 2004). Due to the unique structure of MD and hydrophobicity, sophisticated pre-treatment and derivative protocols are needed prior to instrumental analysis to decrease matrix effect (Wang et al, 2008). Additionally, these instrumental methods are time-consuming, costly and professional, which are not suitable for screening a large number of samples though they are sensitive and accurate (Ha, Song, Ai, & Li, 2016).…”

“…To the best of our knowledge, only few reports focused on the detection of MD using immunoassay methods (Chen, Tang, Ding, He, & Xiao, 2009;Guo, Xu, Song, Liu, & Kuang, 2017;Kennedy, Blanchflower, & O'Dornan, 1997;Shen, Qian, Jiang, & Yang, 2001;Wang et al, 2008). Enzyme-linked immunosorbent assay (ELISA) is low cost, high-throughput and ready-to-use, which have been widely used for screening large samples for food safety Zhu, Song, Liu, Kuang, & Xu, 2016).…”

Monoclonal Antibody-Based Immunoassay for the Detection of Maduramicin in Chicken Tissues

“…In order to monitor MD levels in biological matrixes, a number of analytical methods, such as high-performance liquid chromatography (HPLC) (Gliddon et al 1988;Markantonatos 1988;Johnson 1989;Asukabe et al 1994;Dubois, Pierret, and Delahaut 2004), and immunoassays (Wong 1990;Kennedy, Blanchflower, and O'Dornan 1997;Shen et al 2001;Wang et al 2008), have been developed. As HPLC methods require expensive instrumentation, highly skilled personnel, and extensive sample clean-up, they are not suitable for routine analysis …”