This study involves isothermal kinetic simulation to evaluate the parameters of inhibition conditions for
Escherichia coli
(
E. coli
) and
Staphylococcus aureus
(
S. aureus
) of high-risk pathogens. This is because the new type of the 2019 novel coronavirus (2019-nCoV) is continuously spreading and the importance of public health issues. Environmental disinfection and personal wearing of masks have become important epidemic prevention measures. Selection of concentration kinetics could be estimated best for
E.
coli
and
S. aureus
of pathogens, 2.74 × 10
4
and 10
5
and 2.44 × 10
4
and 10
5
colony-forming units (CFU mL
–1
), by isothermal micro-calorimeter (TAM Air) tests, respectively. Comparisons were made of different doses of 0–70 ppm (in 20 mL test ampoule) hypochlorous acid treatment for conducting
n
th-order and autocatalytic reaction simulation to evaluate the inhibition reaction parameters, which determined the autocatalytic kinetic model that was beneficially applied on the
E. coli
and
S. aureus
. We developed the inhibition reaction parameters of the pathogens, which included the activation energy (
E
a
), the natural logarithm of pre-exponential factor (ln
k
0
), the enthalpy of inhibition microbial growth reaction (
∆H
), inhibition microbial growth, and the inhibition growth analysis. Overall, we conducted isothermal kinetic simulation to understand the antimicrobial activity effects of electrolytically generated hypochlorous acid-treated pathogenic microorganisms, which will provide reference for public health and medical-related fields for SDG3, and can contribute to ensuring human health and hygiene.