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Original ArticlePharmacognosy MagazineVol. 13 | Issue 52s | 2018 | pp. S845–S851Open access
A New Highly Selective and Specific Anti‑puerarin polyclonal Antibody for Determination of Puerarin Using a Mannich Reaction Hapten Conjugate
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- 1 Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University.
- 2 Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology, National Research University, Khon Kaen University, Khon Kaen 40002, Thailand.
- 3 Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812‑8582, Japan.
Published in Pharmacognosy Magazine
Correspondence: Waraporn Putalun
Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology, National Research University, Khon Kaen University, Khon Kaen 40002, Thailand.; Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812‑8582, Japan.
Email: waraporn@kku.ac.th
Copyright: © 2018 Manuscript Technomedia. This is an open access article.
- Published:
- Jan 31, 2018
- Received:
- Jun 27, 2017
- DOI:
- 10.4103/pm.pm_276_17
How to cite
Udomsin, O., Krittanai, S., Kitisripanya, T., Tanaka, H., & Putalun, W. (2018). A New Highly Selective and Specific Anti‑puerarin polyclonal Antibody for Determination of Puerarin Using a Mannich Reaction Hapten Conjugate. Pharmacognosy Magazine, 13(52s), S845–S851. https://doi.org/10.4103/pm.pm_276_17
Abstract
Background: Puerarin (PUE) is a phytoestrogen found in Pueraria candollei and Pueraria lobata. These plants are substantial for traditional medicine in various Asian countries. PUE is a key marker that can be found only in the Pueraria species. Objective: To establish the method for determination of PUE content which is required for quality control of pharmaceutical products. Materials and Methods: PUE‑cationized bovine serum albumin conjugate was created via Mannich reaction. After the rabbit immunization, the obtain anti‑PUE polyclonal antibody (PAb) was used to develop an enzyme‑linked immunosorbent assay (ELISA). Results: An anti‑PUE PAb possess a great sensitivity and specificity. The cross‑reactivity analysis shows no cross‑reaction of an established antibody against other substances. In addition, we successfully developed an indirect competitive ELISA (icELISA) for the quantitative analysis of PUE. The result of method validation conforms to acceptance criteria and correlates with high‑performance liquid chromatography, the reference method. The icELISA was applied to determine PUE content in Pueraria spp. plant samples and its derived pharmaceutical products. Conclusion: This highly specific immunogen was created from the Mannich reaction. An icELISA can also be applied to other research propose in the further studies.
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Article metadata
| Title | A New Highly Selective and Specific Anti‑puerarin polyclonal Antibody for Determination of Puerarin Using a Mannich Reaction Hapten Conjugate |
|---|---|
| Authors | Orapin Udomsin; Supaluk Krittanai; Tharita Kitisripanya; Hiroyuki Tanaka; Waraporn Putalun |
| Affiliations | Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen University.; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology, National Research University, Khon Kaen University, Khon Kaen 40002, Thailand.; Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812‑8582, Japan. |
| Corresponding author | waraporn@kku.ac.th |
| Journal | Pharmacognosy Magazine |
| Volume / Issue | Vol. 13, Issue 52s (2018) |
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