Quality assessment method for Chinpi, dried Citrus spp. peel and its derived Kampo medicines using specific monoclonal antibody against hesperidin
Kanta Noguchi, Daisuke Imahori, Yusuke Kido, Poomraphie Nuntawong, Hiroyuki Tanaka, Satoshi Morimoto, Seiichi Sakamoto- Complementary and alternative medicine
- Drug Discovery
- Plant Science
- Molecular Medicine
- General Medicine
- Biochemistry
- Food Science
- Analytical Chemistry
Abstract
Introduction
Hesperidin (hesperetin 7‐rutinoside, HP), a flavonoid glycoside found in Citrus unshiu Marcowicz or
Objectives
To produce a specific monoclonal antibody against HP (mAb 5D12) to detect the HP contents in Japanese traditional medicines via indirect competitive enzyme‐linked immunosorbent assay (icELISA).
Method
BALB/c mice were immunised with many haptens of HP–bovine serum albumin (BSA) conjugates that were prepared using sodium periodate (NaIO4) to cause an immune response. In addition, conventional hybridoma techniques were utilised to generate mAb 5D12.
Results
The detection range of HP by the mAb 5D12‐based icELISA was 1.56–25.0 ng/mL, with a detection limit of 1.12 ng/mL. The maximum coefficient of variation, as evaluated from the intra‐ and inter‐assays, was <10.0%, and the percentages of recovery, as determined by the spike‐recovery tests, were 105%–115%. Moreover, the HP content, which was obtained from the developed icELISA, correlated well with that obtained via high‐performance liquid chromatography–ultraviolet (HPLC‐UV).
Conclusion
These validation analyses revealed that the established icELISA technique exhibited high precision and accuracy. Notably, this is the first report on the development of icELISA for the HP content‐based quality control of Chinpi and its derived Kampo medicines.