Add time:07/27/2019         Source:sciencedirect.com

Two approaches to set up a functional protein chip platform for the detection of enrofloxacin in milk have been carried out: (A) the functionalization of enrofloxacin with either large carrier proteins (BSA, ALP, KLH, HRP) or bifunctional crosslinkers (adipic acid dihydrazide) and subsequent immobilization on a reactive surface and (B) the chemical surface modification with e.g. adipic acid dihydrazide for covalent binding of unmodified enrofloxacin. The results show that no clear preference can be attributed to either probe- (strategy A) or surface modification (strategy B), as the immobilization efficiency strongly depends on the properties of the selected materials and reagents. Loading capacity is most likely the most important influence factor on the assay performance (besides the introduction of a spacer). Broad dynamic range, high signal intensities and steep slopes were obtained by immobilization of enrofloxacin–enzyme conjugate on epoxy surface and unmodified enrofloxacin on poly-l-lysine surface. Using a KLH-conjugate (A) and the activated poly-l-lysine surface (B) the MRLs of interest (50–100 μg/L) were reached in skimmed milk as well as in unskimmed milk.

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