Through the fluorescence and ultravioletvisible spectrophotometry we have studied the action of cefoperazone sodium and worfloxacin, two kinds of medicinewith albumin, and examined the equal efficiency quality offluorescence enhancement and fluorescence quenching equation. The result shows the equal efficiency of calculating dissociation constant. In this article, we obtain the dissociationconstants of the action of the two kinds of medicine with bovine serum albumin, energy- transfer efficiency, the distanceof donor-acceptor and quenching constants. The above showsthat the action is not caused by dynamic collision but by thebinding of medicine and albumin. This article speciallypoints out that the [Mt]/[Pt], which is the ratio of the maximum mol concentration of the quenching agent (medicine) tofluorescent (albumin) mol concentration when the quenchingends, is very important because it is the maximum bindingnumber of quenching agent molecule to a fluorescent molecule. There is no more quenching after adding more quenching agent beyond this quantity to fluorescent solution. Thisquantity is determined directly and strictly by experiment, sois also of important significance when discussing quenchingefficiency and binding situation of donor-acceptor.