US 8,871,807 B2  58  57  TABLE 11  TABLE 13  TABLE 12  TABLE 14  The test method followed was according to European Stan› dard EN 13704:  Quantitative Suspension Test for the Evalu› ation of Sporicidal Activity of Chemical Disinfectants and  Antiseptics Used in Food, Industrial, Domestic and Institu› tional Areas.  Generally, a test suspension of bacterial spores  in a solution of interfering substance, simulating clean con› ditions, was added to a prepared sample of the test formula› tion diluted in hard water. The mixture was maintained at the  specific temperature and time desired. At this contact time, an  aliquot is taken; the sporicidal action in this portion was  immediately neutralized or suppressed by a validated  method. The number of surviving bacterial spores in each  sample was determined and the reduction in viable counts  was calculated.  The disinfectant properties of each of the formulations at 5  minutes at 40°  C. are shown below in Table 12.  As can be seen from this table, the compositions of the  present invention that were tested were as effective as a dis› infectant as the commercially available formulations tested.  Further, with respect to  Clostridium difficile,  the composi› tions of the present invention were more effective than the  commercially available products tested.  A test was run to determine the stability and sporicidal  efficacy of a composition of the present invention against  spores. The composition tested included the sulfonated per› oleic acid product, and an amount of peroxyoctanoic acid.  The test method used was the European Standard EN 13704:  Quantitative Suspension Test for the Evaluation ofSporicidal  Activity of Chemical Disinfectants and Antiseptics Used in  Food, Industrial, Domestic and Institutional Areas,  described  above. The table below shows the results of this study.  As can be seen from this table, the samples treated with  both a composition of the present invention including the  sulfonated peroleic acid product, and POOA had a higher log  reduction of  Mycobacterium bovis  BCG than those samples  treated with either the sulfonated peroleic acid product or  POOA alone. Although it was found that the samples treated  (b) Stability and Sporicidal Efficacy at 14 Days  For this study, the MS103: Quantitative Tuberculocidal  Test was used. The sulfonated peroleic acid product was  tested alone, and in combination with peroxyoctanoic acid at  various concentrations against  Mycobacterium bovis  BCG.  The compositions were tested at a pH of 6.5 at room tempera› ture. The results are shown in the table below.  ( d) Synergistic Effect of a Compound of the Present Inven› tion with Peroxyoctanoic Acid  The POAA solution alone did not display complete inac› tivation of Poliovirus Type 1 after an exposure time of four  minutes. The reductions in viral titer were s0.75 and s0.50  log 10. When the POAA solution was tested with 1000 ppm of  the sulfonated peroleic acid product, the solution displayed  complete inactivation of Poliovirus Type 1 after an exposure  time of a few minutes, and was therefore efficacious against  the virus. The reduction in viral titer was ;;:.5.75 log 10.  For this study, the ASME 1052-96: Standard Test Method  for Efficacy of Antimicrobial Agents against Viruses in Sus› pension was used. A composition including 1000 ppm per› oxyacetic acid (POAA) was tested alone, and in combination  with sulfonated peroleic acid product.  ( c) Synergistic Effect of a Composition of the Present  Invention with a Known Sanitizes  FIG.  5  shows the stability impact that the compound of the  present invention used, i.e., the sulfonated peroleic acid prod› uct, had on the amount of POOA over time during this study.  As can be seen from this figure, the amount of POOA avail- able over time was higher with the sample of POOA that was  stabilized using a composition of the present invention, com› pared to a sample of POOA that was not stabilized using a  composition of the present invention.  A composition including 30 ppm peroxyoctanoic acid was  also tested. The composition of peroxyoctanoic acid alone did  not result in a reduction.  Test Use Solution  Desired  (Volume of Test  Test  Concentration of  Substance/Total  Formulation  Active Agent  Diluent Volume)  Formula A  1190 ppm  Sterile  0.194 g FormulaA +  witb  MilliQ  170 µL TE/100 g  surfactant  Water  Formula A  1190 ppm  0.194 g FormulaA/  without  100 g  Surfactant  PAPfi  1820 ppm  0.182 g PAP+ 1.5 g  TE/100 g  Ozonitfi  2000  0.200 g Ozonit + 1.5 g  TE/100 g  pH  7.59  8.53  8.50  7.21  Sulfonated Peroleic Acid Product+ POOA  (14 Day Retention)  B. subtilis  60min  DI Water  pH 6.5  Clean Conditions  20°  C.  C.  difficile  60min  Log reduction 3.84  Log reduction 2.71  Log  Test Substance  Exposure Time  Reduction  1000 ppm Sulfonated Peroleic  2.5 min  4.46  Acid Product  5min  5.11  300ppmPOOA  2.5 min  3.48  5min  <4.31  1000 ppm Sulfonated Peroleic  2.5 min  >7.31  Acid Product and  5min  >7.31  300ppmPOOA  1000 ppm Sulfonated Peroleic  2.5 min  a,7.31  Acid Product and  5min  >7.31  150 ppm POOA  FormulaA  Formula A  witb  without  Test/System  Surfactant  PAP  Ozonit  Surfactant  MRSA  >6.66  >6.66  >6.66  >6.66  Enterococcus  >6.26  >6.26  >6.26  >6.26  hirae  ATCC  10541  Escherichia  >6.74  >6.74  >6.74  >6.74  coliATCC  11229  Pseudomonas  >6.32  >6.32  >6.32  >6.32  aeruginosa  ATCC 15442  Clostridium  >3.87  1.17  2.57  3.09  difficileATCC  9689 