A experimental criticism about four-hours method as bioassay of digitalis

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The 4-hours method by frogs as bioassay of digitalis was formally adopted on Pharmacopoeia Japonica VI (1951). This method indicates to give each group of 3 frogs each dose, which has the difference of 20 % (secondly 10%) to successive dose, but this way is not so adequate experimentally or theoretically. We gave to frogs (Rana nigromaculata) subcutaneously 3.2 % solution extracted from many different digitalis leaves by methanol and investigated the time needed to produce the heart standstill. We found from the above results interpolated from 800 cases that all deviations of the time needed to produce the action express completely the normal distribution and that Gaddum's standard deviation, λ is±0.155, 2λ=±0.31. As the products of time and dose are mostly constant in the above dose, the deviation of time should represent that of sensitivity of frogs to digitalis. Calculating the error by the above values of the deviation, the 2ε of determination of digitalis by 4-hours method is about 25 % to the true value of the standard preparation. If we would calculate the error by the method which assigned to give each group of 5 frogs a certain dose, which has the difference of 10 % between each dose, adopted on Deutsches Arzneibuch, it is ± 20% to the former and ±24% to the latter.Even in sucha case that the frog number of one group was more than 5 and the difference between each dose was smaller than 10 %, we could not find the lower value of the error than 15 %.When frogs are few, we can make the error small, if we compute the mean between the median culculated by Kärber's method and the geometrical median of two doses, the one is the large dose that frogs dies almost and the other is the smaller dose that they tolerated almost.
社団法人日本薬理学会の論文

A experimental criticism about four-hours method as bioassay of digitalis

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