On the Estimation of Free Diphtheria Toxin: with reference to the relations existing between lethal doses, lethal times and loss in weight of the guinea-pig

Abstract

1. The relation between lethal doses of diphtheria toxin and the times in which they kill guinea-pigs, or lethal times, is approximately such that the lethal dose multiplied by the corresponding lethal time gives a constant value—a hyperbolic relation.2. Deductions with regard to lethal doses from deaths occurring on the first day or on any day after the sixth give such widely divergent results that, in our opinion, they are at present of negligible value.3. Under the ordinary circumstances of standardisation of diphtheria toxin and antitoxin the relation given in conclusion No. 1 is as close an approximation as any alternative likely to prove of practical utility.4. The graphic representation of L+ doses against lethal times gives a straight line relationship.5. Five of the best investigated toxins confirmed Ehrlich's views in so far as that with toxicities diminishing by 50% their neutralising powers remained practically unimpaired.6. The individual sensitivities of guinea-pigs to free diphtheria toxin render any general relation between lethal times and doses of little value when a small number, e.g. 5, of test animals is inoculated.7. Even with a dose of toxin which would, in any ordinary series of tests, be regarded as an L+ dose, 5 out of 33 guinea-pigs survived the sixth day or 15%1, and the m.l.d. of a well-investigated toxin showed similar variations; consequently both the L+ and m.l.d. values of a toxin are not those amounts which kill with certainty in a fixed time, but those which will cause death in that time with the greatest probability.8. After inoculation, guinea-pigs unless suffering from the consequent “shock,” continue to increase in weight throughout the first day, even with doses that may subsequently prove lethal, and with highly toxic doses the variation in weight during this time is of little significance.9. The greater the amount of free toxin injected, the more rapid is the increment in weight followed by a decrement.10. The weight of a test animal at the end of 24 hours after inoculation forms a better normal or origin with which to compare the subsequent time-weight relations than the initial weight before injection.11. On the basis of Conclusion 10 the general connection is found that with increasing amounts of free toxin the position of the maximum loss of weight is gradually transferred from the second to the fifth day for guinea-pigs which survive over six days.12. The time-weight ratios for almost certainly lethal doses give straight line connections, corresponding probably to starvation curves of the guinea-pig.