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Background  Our previous research showed that icariin ( 1 ) and its phosphorylated structural modification ( 2 ) improved the survival and attenuated oxidative stress and liver dysfunction induced by duck virus hepatitis. In this paper, we were one step closer to determine the structure of phosphorylation icariin ( 2 ) by the FT-IR, HRESIMS and  13 C NMR. Anti-DHAV activities of  1  and  2  were compared in duck embryonic hepatocytes (DEHs) cultured  in vitro  and by artificial infection method  in vivo . Additionally, the antiviral mechanisms of replication/release  in vitro  and the DHAV gene expression  in vivo  of  1  and  2  were analyzed.    Results  Compound  2 's molecular formula was C 33 H 42 O 18 P. The results indicated that  1  and  2  effectively resisted DHAV invading DEHs, that they decreased the mortality of ducklings challenged with DHAV, and that  2  performed more effectively.  1  and  2  performed evenly on DHAV release; however,  2  restrained virus replication far more effectively. Since the anti-DHAV mechanisms of  1  and  2  in vitro  probably involve suppression of replication and release,  2 ’s better performance in anti-DHAV may result from its far more effectively inhibiting virus replication.    Conclusions  The compound  2 's chemical structure was defined as 8-prenylkaempferol-4'-methylether-3-rhamnosyl-7-(6'''-phosphate)-glycoside.  1  and  2  exhibited anti-virus activity on DHAV. Our results suggest that  1  and  2  might become an anti-virus plant material candidate.

Determine the structure of phosphorylated modification of icariin and its antiviral activity against duck hepatitis virus A