Production of Polyclonal Antibody Against Madecassoside and
Development of Immunoassay Methods for Analysis of Triterpene Glycosides
in Centella asiatica.
Centella asiatica (L.) Urban consists of two
major triterpene glycosides, asiaticoside (AS) and madecassoside (MA),
as active components used for wound healing and enhancing memory.
OBJECTIVE=
To
produce a polyclonal antibody against madecassoside (MA-PAb) and
develop enzyme-linked immunosorbent assay (ELISA) and Eastern blotting
methods for quantitative analysis of triterpene glycosides in Centella
asiatica.
METHODS=
An ELISA method was developed using
polyclonal antibody against MA. An Eastern blotting method on the PES
membrane was established for determination of MA and AS. The
immunoassays were validated for sensitivity, precision, specificity and
accuracy.
RESULTS=
The prepared MA-PAb shows specificity
to MA and AS. The measuring range of triterpene glycosides was
0.39-50 µg/mL using the ELISA method. An Eastern blotting method was
developed for determining individual MA and AS, which could be detected
in the range of 62.5-500 ng. The limit of detection for MA and AS was
31.25 ng. The two methods developed showed good specificity, precision,
and accuracy, and also correlated with high-performance liquid
chromatography.