June 8: 9 a. m., reflexes about the same as 5 p. m. previous day, no urine passed since 4.45 p. m. previous day, considerable reduction of Fehling's solution, much more than urine of guinea pig No. 127; 11.05 a. m., convulsions; 12 noon, still alive and stretched out on abdomen; died at 1 p. m. Autopsy: Lungs badly congested; heart and blood vessels injected; blood vessels of kidney and of small intestines injected; liver engorged with blood; a few necrotic spots in stomach.

Guinea pig 127. White, black, and brown male. Weight, 760 grams. Diet, oats and hay.

June 7: 9.50 a. m., 7.6 cc 2 per cent caffein by mouth through stomach tube; 10 a. m., no symptoms; 11.10 a. m., reflexes increased, no urine passed; 1 p. m., very sensitive; 4 p. m., sensitiveness increased, about 20 cc urine passed; 5 p. m., no change.

June 8: 9 a. m., reflexes about the same as 5 p. m. previous day; 9.30 a. m., guinea pig passed 30 cc urine since he received caffein, urine showed a moderate amount of reduction; 12 noon, convulsions; died at 2.30 p. m. Autopsy: Lungs congested; blood vessels of heart and of intestines injected; numerous necrotic spots in stomach; other organs apparently normal.

Examination of the protocols shows that the absorption of caffein from the gastro-intestinal canal was quite rapid, symptoms having been observed as early as 20 minutes after its introduction. The duration of life, it will be remarked, varied with the size of the dose. When approximately 3 decigrams per kilo were fed, all the animals died in the night. They lived, therefore, less than 18 hours. Two decigrams per kilo were likewise fatal, but the duration of life was longer. To decide whether or not this is the smallest fatal dose, smaller amounts were fed. It seemed at first that about 150 mg per kilo was the smallest toxic dose, and about 200 mg per kilo the minimum fatal dose. Macroscopic examination of the organs, however, threw some doubt on this supposition, for well-marked lesions were noticed in all of the guinea pigs which received 0.2 gram per kilo. It is quite possible, therefore, that the minimum fatal dose may be somewhat higher, as we have reason to believe that, at least in some pathologic conditions, the susceptibility to caffein is increased. The presence of fatty changes in the liver of No. 128 and the rapid death in this case lends especial support to this view. Hence, the minimum fatal dose is probably greater than 0.2 gram per kilo for the normal guinea pig. The doses employed for the tests on guinea pigs Nos. 129, 130, and 131 may be considered therefore the minimum fatal dose for these animals. It will be also remarked that macroscopical examination of the organs of these animals failed to reveal the presence of severe lesions. That the minimum toxic dose is probably much smaller than 0.28 gram per kilo is indicated by the experiments on guinea pigs Nos. 135 and 134, in which 0.15 gram caffein per kilo induced mild symptoms in from two to three hours. Both of these, however, and also No. 137 died four to six days after the drug was fed. As already pointed out, caffein may be a factor in the delayed death of guinea pigs which received moderate doses of it. That this supposition may also be true for guinea pigs Nos. 134, 135, and 137 is indeed made probable by the observation that after moderate amounts of caffein symptoms may persist in the guinea pig for about 24 hours, and also by the fact that the secretion of urine in these animals was very scanty, as shown in the preceding record of the experiments; this means slow elimination of caffein and its products of decomposition. It is conceivable that the presence of toxic amounts of caffein in the body for a considerable length of time would induce changes that ultimately lead to the death of the animal or that morbid processes are set up by the combined action of caffein and some preexisting poison. Since some guinea pigs, however, survived the doses indicated, it is more probable that such changes would be brought about by caffein in the presence of a preexisting poison. The death of these pigs, and also of No. 137 several days later, is difficult to account for on any other theory than the one suggested. Were it not for the fact that controls, that is, animals of the same lot which had not received caffein survived all of the experimental animals, changed conditions of environment or accident might be considered the cause of death in the guinea pigs of the last series.

Table 6.—Subcutaneous injection of guinea pigs.

SERIES A.
Number of pig. Weight.Caffein per kiloAppearance of symptoms inDuration of life. Diet. Month. Remarks.
Grams. Mg.
18 500 0.300 2 minutes2 hours 40 minutes CarrotsMarchFemale.
15 548 .29030 minutes Oats do. Do.
14 442 .360 15 minutes do. do. do. Do.
SERIES B.
20 497 0.2002 hours 20 minutes3 hours 30 minutes OatsAprilFemale.
38 570 .210 25 minutes2 hours 15 minutes CarrotsFebruaryMale.
37 820 .2001 hour 25 minutesLess than 18 hoursCarrots and oats. do. Do.
13 618 .19425 hoursOatsMarchFemale.
36 850 .200 1 hour 30 minutes 18 hoursCarrots and oats.FebruaryMale.
SERIES C.
66 510 0.200None5 daysOatsOctoberMale.
65 510 .200 do.Survived do. do. do.
60 320 .2191 hour 15 minutesWithin 18 hours do. do.Female.
57 350 .200 do.About 2½ days do. do. Do.
68 785 .200 25 minutesLess than 22 hours do. do.Male.
69 585 .200 20 minutes9 days do. do. Do.
61 330 .2401 hour 30 minutesLess than 24 hours do. do.Female.
62 335 .2381 hour do. do. do. Do.
70 545 .238 50 minutesAbout 2 days do. do.Male.
71 540 .240 45 minutes do. do. do. Do.
72 560 .232 35 minutesAbout 3 days do. do. Do.
SERIES D.
49 510 0.1601 hour 40 minutesLess than 18 hoursOatsMarchMale.
40 630 .158 Less than 2 daysOats and carrots.February Do.
45 435 .160 None SurvivedOatsMarchFemale.
39 820 .150 2 daysOats and carrots.FebruaryMale.
41 660 .150 Survived do. do.
46 470 .1701 hour 20 minutes do.Oats (?)MarchFemale.
SERIES E.
19 556 0.100Survived OatsApril
42 490 .120 None do. do.February
43 430 .116 do. do. do. do.
44 535 .112 do. do. do. do.
97 330 .100 do. do. do.November
98 520 .100 do.About 3 days Carrots do.
SERIES A.
Number of pig. Weight.Caffein per kiloAppearance of symptoms in
Grams. Mg.
18 500 0.300 2 minutes
15 548 .290
14 442 .360 15 minutes
Number of pig.Duration of life. Diet. Month. Remarks.
182 hours 40 minutes CarrotsMarchFemale.
1530 minutes Oats do. Do.
14 do. do. do. Do.
SERIES B.
Number of pig. Weight.Caffein per kiloAppearance of symptoms in
Grams. Mg.
20 497 0.2002 hours 20 minutes
38 570 .210 25 minutes
37 820 .2001 hour 25 minutes
13 618 .19425 hours
36 850 .200 1 hour 30 minutes
Number of pig.Duration of life. Diet. Month. Remarks.
203 hours 30 minutes OatsAprilFemale.
382 hours 15 minutes CarrotsFebruaryMale.
37Less than 18 hoursCarrots and oats. do. Do.
13OatsMarchFemale.
36 18 hoursCarrots and oats.FebruaryMale.
SERIES C.
Number of pig. Weight.Caffein per kiloAppearance of symptoms in
Grams. Mg.
66 510 0.200None
65 510 .200 do.
60 320 .2191 hour 15 minutes
57 350 .200 do.
68 785 .200 25 minutes
69 585 .200 20 minutes
61 330 .2401 hour 30 minutes
62 335 .2381 hour
70 545 .238 50 minutes
71 540 .240 45 minutes
72 560 .232 35 minutes
Number of pig.Duration of life. Diet. Month. Remarks.
665 daysOatsOctoberMale.
65Survived do. do. do.
60Within 18 hours do. do.Female.
57About 2½ days do. do. Do.
68Less than 22 hours do. do.Male.
699 days do. do. Do.
61Less than 24 hours do. do.Female.
62 do. do. do. Do.
70About 2 days do. do.Male.
71 do. do. do. Do.
72About 3 days do. do. Do.
SERIES D.
Number of pig. Weight.Caffein per kiloAppearance of symptoms in
Grams. Mg.
49 510 0.1601 hour 40 minutes
40 630 .158
45 435 .160 None
39 820 .150
41 660 .150
46 470 .1701 hour 20 minutes
Number of pig.Duration of life. Diet. Month. Remarks.
49Less than 18 hoursOatsMarchMale.
40 Less than 2 daysOats and carrots.February Do.
45 SurvivedOatsMarchFemale.
39 2 daysOats and carrots.FebruaryMale.
41 Survived do. do.
46 do.Oats (?)MarchFemale.
SERIES E.
Number of pig. Weight.Caffein per kiloAppearance of symptoms in
Grams. Mg.
19 556 0.100
42 490 .120 None
43 430 .116 do.
44 535 .112 do.
97 330 .100 do.
98 520 .100 do.
Number of pig.Duration of life. Diet. Month. Remarks.
19Survived OatsApril
42 do. do.February
43 do. do. do.
44 do. do. do.
97 do. do.November
98About 3 days Carrots do.

Table 7.—Injection into peritoneal cavity; guinea pigs.