5-sulfosalicylic Acid Dihydrate-Based Pretreatment for the Modification of Enzyme-Linked Immunoassay of Fluoroquinolones in Fishery Products

Abstract

A simple, rapid sample extraction method for the determination of FQs was developed. Fishery samples were extracted with 2% of 5-sulfosalicylic acid dihydrate and the extracts were analyzed directly without any further purification or clean-up procedures. The FQs were determined with standards of 2% of 5-sulfosalicylic acid dihydrate in the concentration range of 0.1-25.6 μg L(-1), and the limit of detection (LOD) was 0.1 μg L(-1). The matrix interference originated from fishery samples was eliminated by 2% of 5-sulfosalicylic acid dihydrate and did not interact with horseradish peroxidase (HRP) labeled IgG in western blotting. No significant matrix interference was observed as samples extracted with 2% of 5-sulfosalicylic acid dihydrate. Recoveries of FQs in fishery muscle were between 72.37-94.35% in the concentrations range of 10-50 μg kg(-1).This extraction procedure was much rapider and simpler to conventional ELISA extraction procedure and could be used as a time-saving and cost-effective method for FQs monitoring in fishery samples.

The effect of fish matrix on colloidal gold immunochromatographic assay of antibiotics: the source of interference and removal method

“…Even after the complete removal of all the proteins, the colour darkness on the test line was still significantly lighter in comparison to that on the control line, which indicated the presence of other sources of matrix effect besides proteins. Such a result was quite different from that observed in the case of ELISA, in which the influence of proteins seemed dominant in comparison to other water-soluble components, and little interference remained after the elimination of proteins by the 5-sulfosalicylic acid treatment (Cui et al, 2015;Qu et al, 2016).…”

“…The crude extracts were further mixed by 4% 5-sulfosalicylic acid dehydrate to a final volume of approximately 2:1 (v/v) of the original volume in order to further remove the protein (Cui et al, 2015). The solution was then centrifuged at 4132 × g for 15 min at 4°C and the supernatants were collected again.…”

“…Recently understanding of the matrix effect on ELISA has made significant progress, for example, some proteins have been verified as the main interference-inducing components in fishery products, and their chemical, physical and biological characteristics, as well as the action mechanism, have been clarified (Wang et al, 2016). Based on these results, new and more effective approaches have been developed for the elimination of the matrix interference by the exploitation of 5-sulfosalicylic acid dehydrate during pre-treatments (Cui, Lin, Wang, Cao, & Sui, 2015;Qu et al, 2016). For the matrix interference to GICA, though the effect of proteins and Na + was proposed in food or urine samples (Li et al, 2013;Xu, 2013), overall the detailed information is still very limited, especially for fishery products and other food samples, which usually have complex components.…”

The effect of chlorophyll on the enzyme‐linked immunosorbent assay (ELISA) of procymidone in vegetables and the way to overcome the matrix interference

“…Among various acids, 5-sulfosalicylic acid has demonstrated excellent ability to eliminate matrix proteins with little effect on the ELISA and other immunoassays. 24,32,33 Considering the excellent stability of procymidone under acidic conditions, in principle 5-sulfosalicylic acid would not react with procymidone or lead to its breakdown. 34 These proposals were confirmed well by the following experiments.…”

Isolation, characterization, and identification of proteins interfering with enzyme-linked immunosorbent assay of antibiotics in fish matrix

“…Low pH values (especially under 5) seemed able to eliminate part of the matrix effects by precipitation of the 42 kD protein, but the interference from the 36 kD protein was still present. Our previous study (25) investigated the effect of 5sulfosalicylic acid dehydrate on these interfering proteins, and they could be evidently precipitated. Moreover, an appropriate concentration of 5-sulfosalicylic acid dehydrate, such as 2%, was demonstrated to have little influence on ELISA performance.…”