Immunoquantitative analysis of artemisinin from Artemisia annua using polyclonal antibodies.

Abstract:

:Artemisinin was derivatized to dihydroartemisinin carboxymethylether in three steps, without disturbing the peroxide bridge, and then linked to either thyroglobulin (TGB) or bovine serum albumin (BSA). The artemisinin-TGB and -BSA conjugates were injected in female New Zealand rabbits but only the artemisinin-TGB conjugate generated polyclonal antibodies. An enzyme-linked immunosorbent assay (ELISA) was developed and the specificity of the antibodies was confirmed by comparison with pre-immune serum and by competitive assays using different dilutions of artemisinin standards. Although anti-artemisinin antibodies cross-reacted with artemisitene and dihydroartemisinin at all dilutions used, cross-reaction with deoxyartemisinin, artemisinic acid, and arteannuin B occurred only at high concentrations. ELISA successfully detected artemisinin from crude extracts in concentrations as low as 1.5 ng ml-1; and was epsilon 400-fold more sensitive than the HPLC-EC. The ELISA successfully detected and quantified artemisinin in different organs of greenhouse-grown plants and in eight clones of Artemisia annua grown in tissue culture but artemisinin was overestimated owing to cross-reactivity of the antibodies with artemisinin-related compounds present in the samples. Despite overestimation of artemisinin content, the correlations between ELISA and HPLC-EC were r = 0.92 when samples were diluted 100 times, and r = 0.90 when samples were diluted 500 times, indicating that ELISA is a potential tool for screening large A. annua populations.

journal_name

Phytochemistry

journal_title

Phytochemistry

authors

Ferreira JF,Janick J

doi

10.1016/0031-9422(95)00542-0

subject

Has Abstract

pub_date

1996-01-01 00:00:00

pages

97-104

issue

1

eissn

0031-9422

issn

1873-3700

pii

0031-9422(95)00542-0

journal_volume

41

pub_type

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