PROTECTING FIBRINOGEN WITH RUTIN DURING UVC IRRADIATION FOR VIRAL INACTIVATION

Fibrinogen solutions were irradiated with WC (254 mn) to inactivate co ntaminating viruses, In order to protect fibrinogen during UVC irradia tion, 0.5 mM rutin was added prior to UVC exposure and subsequently re moved during processing, Viral kill by 0.1 J/cm(2) UVC resulted in the following inactivation values (log 10): non-lipid-enveloped viruses: Parvo greater than or equal to 5.5; encephalomyocarditis virus greater than or equal to 6.5; hepatitis A virus greater than or equal to 6.5: lipid-enveloped viruses: human immunodeficiency virus greater than or equal to 5.7; vesicular stomatitis virus greater than or equal to 5.7 . Fibrinogen irradiated with 0.5 mM rutin did not significantly differ from unirradiated material in terms of clot time and breaking strengt h, In the absence of rutin, UVC irradiation of fibrinogen at similar f luence led to loss of solubility, increased clot time and the cleavage of fibrino-peptides that reacted with dinitrophenyl hydrazine as a te st for ketonic carbonyl groups. High-performance liquid chromatography and mass spectrometry data showed that rutin exposed to UVC formed nu merous breakdown, oxidation and combinational products, Experiments wi th H-3-rutin showed that after UVC irradiation, subsequent processing by a C18 resin and alcohol precipitation removed >99% rutin, represent ing <10 ppm rutin in the final fibrinogen preparations, Residual H-3-r utin was not covalently bonded to the fibrinogen, Immunochemical studi es with rabbit antisera to UVC irradiated (with rutin) fibrinogen show ed the absence of neoimmungens. By all measures, rutin prevents fibrin ogen degradation during virucidal UVC irradiation.