ABSTRACT
<p><b>OBJECTIVE</b>To inspect the quality of commercially available Chaihu (Bupluri Radix, a common traditional Chinese herbal drug) in Guangzhou by determining the content of saikosaponin A (SSa) using enzyme-linked immunosorbent assay (ELISA).</p><p><b>METHODS</b>A competitive ELISA system using mouse anti-saikosaponin A monoclonal antibody was established for determining SSa content. Commercial samples of Chaihu were obtained from 10 drug stores in Guangzhou, and SSa contents in the methanol extracts of these samples were determined using the ELISA system.</p><p><b>RESULTS</b>The detection range of this competitive assay was 0.16-2.5 µg/ml for determining SSa contents. In the 10 commercial Chaihu samples, SSa contents in the methanol extract determined by this method ranged from 0.32 µg/mg to 6.87 µg/mg, and 3 samples showed a SSa content lower than the minimum requirement documented in the Chinese Pharmacopeia.</p><p><b>CONCLUSION</b>This competitive ELISA is sensitive, rapid, economic and environment-friendly for SSa determination, especially suitable for batch determination. The results of SSa detection for the commercial Chaihu samples demonstrate an uneven quality of Chaihu in Guangzhou market, suggesting the necessity of more rigorous quality control measures for this drug.</p>
Subject(s)
Antibodies, Monoclonal , Bupleurum , Chemistry , Drugs, Chinese Herbal , Enzyme-Linked Immunosorbent Assay , Methods , Oleanolic Acid , Quality Control , SaponinsABSTRACT
<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 (25:75:1). 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>