Few assumptions in the calculation of ATP molecules are -

(a) It is assumed that various parts of aerobic respiration such as glycolysis, TCA cycle, and ETS occur in a sequential and orderly pathway.

(b) NADH produced during the process of glycolysis enters into mitochondria to undergo oxidative phosphorylation.

(c) Glucose molecule is assumed to be the only substrate while it is assumed that no other molecule enters the pathway at intermediate stages.

(d) The intermediates produced during respiration are not utilized in any other process.