Synthesis of haptens and gold-based immunochromatographic paper sensor for vitamin B6 in energy drinks and dietary supplements
Lu Zeng, Xinxin Xu, Shanshan Song, Liguang Xu, Liqiang Liu, Jing Xiao, Chuanlai Xu, Hua Kuang
Abstract
We designed and synthesized novel haptens to produce monoclonal antibodies (mAb) against vitamin B6 (VB6). A group-specific mAb (2F9) that recognized pyridoxine (PN), pyridoxamine (PM), and pyridoxal (PL) was prepared using a homologous strategy with 50% maximal inhibitory concentration (IC50) values of 106.60, 250.57, and 400.11 ng/mL, respectively. Based on this, a gold nanoparticles (AuNPs)-based immunochromatographic strip (ICS) test was established for the detection of VB6 in energy drinks and B-vitamin complex tablets. The developed ICS test results could be semi-quantitatively evaluated by the naked eye within 10 min, and displayed the visual limit of detection (vLOD) values of 250, 500, and 1,000 ng/mL for PN, PM, and PL, respectively. For quantitative analysis, the results obtained by strip reader, with calculated LOD values for PN, PM, and PL were 14.10, 55.58, and 56.25 ng/mL, respectively. Commercial energy drinks and B-vitamin complex tablet samples were detected by the strips and the results were confirmed with high-performance liquid chromatography. Overall, the developed AuNPs-based immunochromatographic sensor was suitable and promising for the group-specific recognition and rapid detection of VB6 in fortified foods and dietary supplements.