Acetylcholinesterase (AChE) is an externally oriented membrane-bound enzyme and its main physiological role is termination of chemical transmission at cholinergic synapses and secretory organs by rapid hydrolysis of the neurotransmitter acetylcholine (ACh). Nevertheless, it is well known that cyclophosphamide (CP; nitrogen mustard derivative) is an eminent anticancer drug. The present work addresses multiple approaches to analyze an identical data for rat brain AChE inhibition by CP. These different angels of analysis based on two classical (Lineweaver-Burk as well as Dixon) plots, their secondary replots, a new graphical approach and general built-in equations of GOSA. Thus various kinetic constants (K(I), K(s,) K(m), k(sl), V(mao), K(i), k(sli), S(lo), K(maxi), S(K0.5), k(cat) and k(sp)) were estimated and mode of inhibition discussed in the current study.