US 8,871,807 B2  62  61  slowly added, producing a clear solution. After sitting at 50°  C. for 24 hours, the solution was analyzed to determine the  presence of a peracid.  To determine the presence of a peracid, a kinetic iodomet- ric titration similar to the method disclosed in Sully and 5  Williams ("The Analysis of Per-Acids and Hydrogen Perox› ide,"  The Analyst,  87: 1037, p. 653 (August 1962)) was used.  This method has demonstrated a lower detection limit of  about 0.3 ppm for POAA. Given the molecular weight ratio of  POAA to the perspective percarboxylic acid of PC-48, the  detection limit was estimated to be about 1.4 ppm (3.93xl0- 6  M). No peracidformation was observed. This is equivalent to  a percarboxylic acid formation constant (Keq) less than  0.002, suggesting substantially no peracid was formed.  Alternatively, formation of the peracid was determined  using $^{13}$C NMR (D$_{2}$O). Using this technique, no carbonyl  resonance signal from the peracid was observed.  Other a-sulfonated fatty acid sources such as Alpha-Step  PC-48 and Alpha-Step BSS-45 were also reacted with H$_{2}$O$_{2 }$ in a similar manner, and in both cases, no corresponding  peracids were detected.  Non-a-sulfonated fatty acids were also tested to determine  the likelihood of peracid formation. For the sulfonated oleic  acid discussed above, the measured formation constant was  1.42. The sulfonated undecenoic acid was collected as a  stable solid powder, so the formation constant was not mea› sured. Although the formation constant of the peracid of  sulfonated oleic acid is significantly lower than that of the  most common commercialized peracid, peroxyacetic acid  (Keq=2.70), it is still high enough to make practical yields.  Overall, without wishing to be bound by any particular  theory, it is thought that the a-sulfa group prohibits the oxi› dation and/or perhydrolysis of the carboxylic acid group by  H$_{2}$O$_{2 }$to the corresponding peracid. This may in part be due to  its strong electron withdrawing effects.  A study was run to determine the efficacy of compositions  of the present invention as sanitizers used in a clean in place  cleaning method. A composition including about 5.85 wt%  of the sulfonated peroleic acid product, and about 11.6%  hydrogen peroxide, about 1 wt % of a chelating agent, about  12.75 wt% ofH$_{2}$SO$_{4}$ , about 13.6 wt% NAS-FAL (sodium  octane sulfonate ), and about 1.5 wt% of SXS ( commercially  available from the Stepan Company) was prepared. Synthetic  hard water was used to dilute the test composition to the  desired peracid concentration. The peracid was tested at con› centrations of 1000 ppm, 750 ppm and 500 ppm. The pH of  the use solutions were as follows:  As can be seen from these results, the use solutions tested  were effective disinfectants against both  Staphylococcus  aureus,  and  Pseudomonas aeruginosa  at the concentrations  tested.  Another study was run to determine the sanitizing efficacy  of the test solution against  Staphylococcus aureus  ATCC  6538 and  Escherichia coli  ATCC 11229 after a 30 second  exposure time. For this experiment the solutions were diluted  isms. The plates were incubated at 35°  C. for 48 hours with  the test systems prior to exposure to the peracids. The results  are shown in the table below.  TABLE 16  Example 9  Clean in Place Sanitizing Compositions  Test Substance  # Negative Tubes/# Carriers Tested  Staphylococcus aureus  ATCC 6538  1000 ppm Peracid Composition  60/60  Pseudomonas aeruginosa  ATCC 15442  1000 ppm Peracid Composition  60/60  Control  Negative Carrier  Positive Carrier  Positive Carrier  Organic Soil  Neutralization  (lO00ppm)  Neutralization  (lO00ppm)  Culture Enwneration  Culture Enwneration  Carrier Enumeration  Carrier Enumeration  Test Substance  Test Controls  Test System  Staphylococcus aureus  ATCC 6538  Results  1 negative of 1 tested  1 positive of 1 tested  Pseudomonas aeruginosa  1  positive of  1  tested  ATCC 15442  1 negative of 1 tested  Staphylococcus aureus  6 positive of 6 tested  ATCC 6538  Pseudomonas aeruginosa  6 positive of 6 tested  ATCC 15442  Staphylococcus aureus  ATCC 6538  9.0 x 10 $^{8 }$CFU/mL  Pseudomonas aeruginosa  1.0 x 10 $^{9 }$CFU/mL  ATCC 15442  Staphylococcus aureus  ATCC 6538  Pseudomonas aeruginosa  ATCC 15442  1.0 x 10 $^{6 }$CFU/mL  1.0 x 10 $^{7 }$CFU/Carrier  2.3 x 10 $^{6 }$CFU/mL  2.3 x 10  $^{7 }$CFU/Carrier  # Negative Tubes/# Carriers Tested  Staphylococcus aureus  ATCC 6538  500 ppm Peracid Composition  59/60  750 ppm Peracid Composition  60/60  Pseudomonas aeruginosa  ATCC 15442  500 ppm Peracid Composition  750 ppm Peracid Composition  58/60  60/60  Control  Negative Carrier  Positive Carrier  Positive Carrier  Organic Soil  Neutralization  Neutralization  Culture Enwneration  Culture Enwneration  Carrier Enumeration  Carrier Enumeration  Test Controls  Test System  Staphylococcus aureus  ATCC 6538  Results  1 negative of 1 tested  1 positive of 1 tested  Pseudomonas aeruginosa  1  positive of  1  tested  ATCC 15442  1 negative of 1 tested  Staphylococcus aureus  3 positive of 3 tested  ATCC 6538  Pseudomonas aeruginosa  3 positive of 3 tested  ATCC 15442  Staphylococcus aureus  ATCC 6538  1.0 x 10 $^{9 }$CFU/mL  Pseudomonas aeruginosa  1.0 x 10 $^{9 }$CFU/mL  ATCC 15442  Staphylococcus aureus  7.2 x 10 $^{5 }$CFU/mL  ATCC 6538  7.2 x 10 $^{6 }$CFU/Carrier  Pseudomonas aeruginosa  2.0 x 10 $^{6 }$CFU/mL  ATCC 15442  2.0 x 10 $^{7 }$CFU/Carrier  Concentration of Peracid in Use Solution  500 ppm peracid  750 ppm  1000 ppm  pH  1.65  1.46  1.38  The use solutions were tested against  Staphylococcus  aureus  ATCC 6538 and  Pseudomonas aeruginosa  ATCC  15442. The organic soil used was 5% Fetal Bovine Serum.  The exposure time of the test was 5 minutes at a temperature  of20–1 °  C. A neutralizer screen was also performed as part of  the testing to verify that the neutralizer adequately neutral› ized the product and was not detrimental to the tested organ-