Disruption of influenza virus A by diethylether-Tween and tri-N-butyl phosphate-Tween mixtures

In search for optimal conditions of influenza virus A/Brazil/78(H1N1) disruption by diethylether-Tween 80 and tri-n-butyl phosphate (TNBP)-Tween 80 mixtures, the following treatments were found suitable: for 120 min at 4 degrees C with 3.3% TNBP and 0.1% Tween 80 or for 120 min at 4 degrees C with diethylether and 0.1% Tween 80 (ratio of diethylether and treated virus material 1:1). Disruption by TNBP appeared more favourable not only because of the convenient performance but also due to the higher antibody-inducing ability of the product obtained. The suggested removal of TNBP from the disruption product by extraction into hexan is easy and reliable. Chemical analysis enabled precise detection of 0.1% TNBP in the "vaccine" product. The hexan-extracted "vaccine" contained less than 0.05% TNBP, a concentration non-toxic for mice.