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An enzyme‐linked immunosorbent assay for the measurement of plasma flavonoids in mice fed apigenin‐ C ‐glycoside
Author(s) -
Ninfali Paolino,
Dominici Sabrina,
Angelino Donato,
Gennari Lorenzo,
Buondelmonte Costantina,
Giorgi Luca
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6143
Subject(s) - vitexin , immunogen , apigenin , bovine serum albumin , chemistry , bioavailability , antigenicity , antiserum , polyclonal antibodies , in vivo , biochemistry , glycoside , hapten , antibody , chromatography , pharmacology , biology , flavonoid , monoclonal antibody , immunology , stereochemistry , microbiology and biotechnology , antioxidant
BACKGROUND In the Chenopodiaceae family, the apigenin flavonoids vitexin‐2‐ O ‐xyloside ( VOX ) and vitexin‐2‐ O ‐rhamnoside ( VOR ) are important chemopreventive components. To investigate their bioavailability in in vivo animal studies an enzyme‐linked immunosorbent assay ( ELISA ) method has been developed . RESULTS The ELISA was based on polyclonal antibodies elicited in mice by injecting, as an immunogen, 4′,6″‐ O ‐biapigenin (hinokiflavone, HF ) conjugated to bovine serum albumin ( BSA–HF ). A second immunogen was synthesised by coupling an equimolar mixture of VOX and VOR to BSA ( BSA–F1 ). The BSA–HF elicited a significant antibody response, due to 17 HF hapten groups, coupled to each BSA molecule, whereas BSA–F1 provided a very low antigenicity in respect to control animals. Antiserum raised against BSA–HF showed an antibody titre of 1:1600. Antibodies were found to be specific for the flavonols. Our results show that VOX and its metabolic products reached the concentration of 3.42 ± 0.72 µg mL −1 in plasma of VOX fed animals, at the net of the control value . CONCLUSIONS By using the ELISA , the concentration of apigenin flavonoids and their metabolites can be detected in VOX ‐ or VOR ‐supplemented animals. The assay represents a useful tool for rapid screening to compare bioavailability of apigenin flavonoids in respect to control animals. © 2013 Society of Chemical Industry