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Development of immunoaffinity chromatography to specifically knockout baicalin from Gegenqinlian Decoction
Author(s) -
Zhao Yan,
Feng Huibin,
Shan Wenchao,
Cheng Jinjun,
Wang Xueqian,
Zhao Yan,
Qu Huihua,
Wang Qingguo
Publication year - 2015
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201500168
Subject(s) - baicalin , chemistry , decoction , baicalein , chromatography , wogonin , affinity chromatography , high performance liquid chromatography , column chromatography , glucuronide , mangiferin , biochemistry , scutellaria baicalensis , traditional medicine , pharmacology , metabolite , medicine , alternative medicine , traditional chinese medicine , pathology , enzyme
Specific knockout technology provides a powerful tool to confirm the role of target compounds in a plant or its derived prescriptions, and this principle is the same as that with knockout genes. In this study, we generated an immunoaffinity column conjugated with an anti‐baicalin monoclonal antibody and then loaded Gegenqinlian Decoction extracts, followed by washing with deionized water and an elution solvent. The results of the high‐performance liquid chromatography fingerprints and high‐performance liquid chromatography with mass spectrometry showed that the immunoaffinity column was able to specifically knockout baicalin, oroxylin A‐7‐ O ‐glucuronide, wogonoside, wogonin, and baicalein from Gegenqinlian Decoction. A reliable one‐step method to specifically knockout baicalin was established with an immunoaffinity column. Gegenqinlian Decoction and its knocked‐out fraction induced the expression of superoxide dismutase and were compared in human umbilical vein endothelial cells cultured with a high glucose concentration; the results showed that the Gegenqinlian Decoction and its knocked‐out fraction showed no significant difference, which indicated that the baicalin, oroxylin A‐7‐ O ‐glucuronide, wogonoside, wogonin, and baicalein that were knocked out by the immunoaffinity column might not be key compounds for the induction of Gegenqinlian Decoction superoxide dismutase secretion.