Preparation and Characterization of an Immunoaffinity Column for the Selective Extraction of Salbutamol from Pork Sample

Abstract

A rapid, simple, and reliable determination method for salbutamol in pork was developed with immunoaffinity column (IAC) extraction followed by HPLC analysis. The salbutamol immunoaffinity column was prepared by coupling CNBr-activated Sepharose-4B with the anti-salbutamol polyclonal antibody which was purified by caprylic acid-ammonium sulfate. The coupling rate of the antibody and Sepharose-4B was 98.6%, and the dynamic column capacity of IAC was 400 ng/mL gel. The average recoveries of salbutamol from spiked pork samples at levels of 2, 10, 20, and 50 ng/g ranged from 83.3% to 92.2%, with the relative standard deviations of 2.8-7.0% (n=5), and the limits of detection and qualification were 0.25 ng/g and 0.5 ng/g, respectively.

Synthesis and application of boronic acid‐functionalized magnetic adsorbent for sensitive analysis of salbutamol residues in pig tissues

“…A number of sample pre‐treatment methods, based on novel adsorbents, have been developed for the analysis of SAL. C 18 ‐SPE (Fiori et al ., ), weak cation exchange columns (Meng et al ., ), molecularly imprinted polymers (Yan et al ., ), polymer microextraction (Huang et al ., ; Du et al ., ), immunoaffinity columns (Wang et al ., ), tert ‐butylmethylether extraction (Garrido et al ., 2013) and poly (MAA‐EGDMA) SPE columns (Wang et al ., ) have been used to extract SAL from the complex biological samples. However, most of these methods are nonspecific and may lead to nonspecific adsorption from the sample matrix.…”

An Ultra-Sensitive Monoclonal Antibody-Based Competitive Enzyme Immunoassay for Sterigmatocystin in Cereal and Oil Products

“…To eliminate the influence of impurities generated in sample extraction on ELISA and improve the sensitivity of this method, the immunoaffinity column (IAC) was prepared as described by Wang et al with some modifications [14]. The procedures were as follows: 10 µg CNBr-activated Sepharose-4B matrix powder was dissolved into 1 mM HCl and then placed into a sintered-glass filter funnel and washed with the HCl solution for 15 min; the CNBr-activated Sepharose-4B was sequentially washed with 25 mL coupling buffer for 5 times, and then quickly transferred into the coupling buffer with 5 mg mAb and thoroughly mixed by stirring (180 rpm for 1 min) at room temperature for 2.5 h; the coupling products were obtained with a sintered-glass filter funnel and washed with the coupling buffer and blocking buffer to remove free ligands, and then transferred into 0.1 M Tris-HCl buffer (pH 8.0) and stirred (180 rpm for 1 min) at room temperature for 2 h. To remove the uncoupled blocking ligands, the coupling products were sequentially washed with 0.1 M HAc-NaAc (pH 4.0) and Tris-HCl buffer for at least 5 cycles.…”

Advances in the Determination of Anabolic-Androgenic Steroids: From Standard Practices to Tailor-Designed Multidisciplinary Approaches

“…Three generations of IAC methods for the extraction of methandienone were developed by Wang et al [ 80 , 81 , 82 ]. Their methodology included immunogen synthesis and gaining polyclonal Abs [ 82 ].…”

“…Three generations of IAC methods for the extraction of methandienone were developed by Wang et al [ 80 , 81 , 82 ]. Their methodology included immunogen synthesis and gaining polyclonal Abs [ 82 ]. Subsequently, a transition to monoclonal Abs followed, which significantly increased the binding capacity of the immunosorbent [ 81 ], while the development of improved chitosan beads led to the homogenization and improved stability of the obtained immunosorbent [ 80 ].…”