Disclosed is a method of assaying the content of Mg ions in a human body fluid such as serum, urine or saliva by the use of a reactant solution containing isocitrate dehydrogenase, NADP.sup.+, isocitrate and an excess amount of a chelating agent. Almost all Mg ions as existing in a human body fluid sample to be examined are bonded with the chelating agent, and the remaining Mg ions react with NADP.sup.+ to form NADPH. Increase of the thus formed NADPH is measured to obtain a standard curve of a straight line, and the content of Mg ions in the same is determined on the basis of the standard curve. If such an excess amount of a Mg-chelating agent is not added to the reactant solution, the intended standard could not be in the form of a straight line but is in the form of a tangent curve. Using the tangent curve, the content of Mg ions in the human body fluid sample cannot be assayed accurately.