Immunodetection of ginsenoside Rb1 in rat serum / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To establish a novel immunoassay for qualitative detection of ginsenoside Rb1 in rat serum.</p><p><b>METHODS</b>Anti-G-Rb1 monoclonal antibody (mAb) was through a hybridoma approach. Rat serum containing G-Rb1 was deproteinized with methanol to prepare the sample for testing, which was loaded onto polyethersulfone (PES) membrane and developed in the mixture of acetonitrile, water and acetic acid (25751). After treatment with NaIO(4), the membrane was transferred to 1% BSA solution for immobilization of G-Rb1. The membrane was subsequently treated with anti-G-Rb1 mAb solution, followed by addition of peroxidase-labeled goat anti-mouse IgG and color development using 4-chloro-1-naphthol-0.03% H(2)O(2).</p><p><b>RESULTS</b>On the PES membrane, a clear blue spot representing G-Rb1 occurred where the rat serum for testing and the standard G-Rb1 samples were blotted. The limit of this immunodetection was 0.25 microg.</p><p><b>CONCLUSION</b>This immunoassay has greater specificity and reliability than thin-layer chromatography with a sensitivity similar to that of high-performance liquid chromatography, and does not require sophisticated equipment for convenient G-Rb1 detection in rat serum.</p>