A New Highly Selective and Specific Anti-puerarin polyclonal Antibody for Determination of Puerarin Using a Mannich Reaction Hapten Conjugate

Pharmacognosy Magazine
Orapin UdomsinWaraporn Putalun

Abstract

Puerarin (PUE) is a phytoestrogen found inPueraria candolleiand Pueraria lobata. These plants are substantial for traditional medicine in various Asian countries. PUE is a key marker that can be found only in thePuerariaspecies. To establish the method for determination of PUE content which is required for quality control of pharmaceutical products. 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). 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 inPuerariaspp. plant samples and its derived pharmaceutical products. This highly specific immunogen was created from the Mannich reaction. An icELISA can also be applied to other research pro...Continue Reading

Methods Mentioned

BETA
enzyme-linked immunosorbent assay
ELISA
icELISA

Software Mentioned

icELISA

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