March, 1933
TOXICITIES OF ROTENONE DERIVATIVES
1147
Anal. Calcd. for C~~HOON*O~: C, 59.96; H,8.39; N,11.66. Found C,59.71;
H,8.36; N,11.00.
stlmmary 1. The behavior of some 2-keto-4-alkyl-5-carbethoxy-6-methyl-1,2,3,4tetrahydropyrimidines on catalytic reduction has been investigated. 2. It has been shown that the 5,6-double bond in such pyrimidine combinations is very resistant to change. 3. In all cases examined the 4-aryl groups were attacked by hydrogen in the presence of the catalyst and reduced to the corresponding hexahydro or saturated structure. 4. In only one case examined, namely, 2-keto-4-methyl-5-carbethoxy6-pheny1-1,2,3,4-pyrimidine, where phenyl is substituted on carbon adjacent to the 5,6-double bond, did we succeed in reducing the 5,6-double bond in the pyrimidine ring. 5. The pharmacological behavior of these highly reduced structures is now under investigation. NEWHAVEN,CONNECTICUT
RECEIVED SEPTEMBER 1, 1932 PUBLISHED MARCH 7, 1933
[CONTRIBUTION FROM THE INSECTICIDE DIVISION, BUREAU OF CmMISTRY AND
SOILS]
A Study of the Toxicity of Rotenone Hydrochloride, Acetylrotenone and Rotenolone Using the Goldfish as the Test Animal' BY W. A. GERSDORFF The toxicological examination of derivatives of rotenone and related compounds with the use of the goldfish as the test animal has been continued in this Laboratory with the threefold hope of discovering a material more toxic and more stable than rotenone, and a t the same time securing data by which a correlation may lie made between the toxicity and chemical structure. The method used by the author has been described in a previous paper2 and studies by that method of some of the compounds have also been This pa.per presents the results of a similar examination of rotenone hydrochloride, acetylrotenone and rotenolone prepared in the Insecticide Division of the Bureau of Chemistry and Soils. Rotenone hydrochloride (m. p. 193') was prepared from rotenone by H. L. Haller according to the method of S. TakeL6 The compound is formed by the addition of hydrochloric acid at the double bond of the (1) Presented before the Division of Agricultural and Food Chemistry at the Meeting of the American Chemical Society, Denver, Colo., August 22-26, 1932. (2) Gersdorff, THISJOURNAL, 68, 3440-3445 (1930). (3) Gersdorff, ibid., 62, 5051-5056 (1930). (4) GersdorB, ibid., 69, 1897-1901 (1931). (5) Takei, Ber., 618, 1003-1007 (1928).
W.A. GERSDORFP
1148
VOl. 55
tubaic acid side chain. Its structure, now that the formula for rotenone has been established,6is shown by the formula
CH ?$ ;YJ
\/ Hz
-0
\
c1
I I
CH3
Hz c-c-c/ )-,
CHsO
P-COCHI
-0
Hz
c--lc-cy Hz H
CHz \CHz
Rotenolone (m. p. 140-141') was prepared* by the saponification of the acetate obtained as one of the products of the reaction of iodine on a hot alcoholic solution of rotenone and potassium acetate. The hydroxyl group replaces the hydrogen attached to one of the adjacent asymmetric carbon atoms of rotenone, as 0
or
Which of these two formulas is correct has not yet been established. The two lots of fishes used in these tests were slightly larger than the fishes used for the determination of the toxicity curves for rotenone and some of its derivatives in 1930,a and apparently somewhat more resistant (6) LaForge and Haller, THISJOURNAL, 64, 810-818 (1932). (7) Smith and LaForge, ibid., 64, 2996-3000 (1932). (8) LaForge and Smith, ibid., 64, 1091-1098 (1930).
htarch, 1933
TOXICITIES OF ROTENONE DERIVATIVES
1149
as shown in Table I. In this, comparisons are made of the three lots of fishes at two concentrations of rotenone. TABLEI RESISTANCE OF THREE LOTSOF GOLDFISHES TO ROTENONE AT 27" Lot no. of Eshes
1
2 3 I 2 3
Concn., mg. per liter
No. of fishes used
Mean wt. of fishes, g.
Mean survival,time, min.
0.10 .10 .10 .05 .05 -05
12 5 8 11 10 12
2.3 2.4 2.4 2.2 2.7 2.3
95 114 123 150 180 177
TABLEI1 TOXICITY OF ROTENONE HYDROCHLORIDE TO GOLDFISH AT 27.0 Concn., mg. per liter
Mean length No. of of fishes, fishes used mm.
41 41 40 41 41 41 42 43 42 42
.0050
9 12 9 12 13 11 9 11 18 7 10
.0025
11
b
0.50 ,33 .20 .17 * 10 .050 .033 .025 .020 .015
b
Mean weight of fishes." g.
Mean eurv. time, min.
* 0.2
O
100 M~~~ ___ surv. time
2.2 2.2 2.1 2.2 2.2 2.2 2.3 2.4 2.3 2.3
128 0.83 125 .89 124 .a 127 .87 130 .82 138 .74 181 .60 212 .51 273 .46 386 .28 Eight fishes, 98OC,two ,12' still active after 72 hrs. Apparently unaffected, 72 hrs.
'Estimated from length. Fishes not measured, but of same approximate size. These figures are only approximate since a large number of fishes would be required to give an accurate mean value. The reciprocal of the survival time of a fish surviving the test is taken as zero, since the reciprocal of any survival time longer than the test would be negligibly small. The rotenone curves were obtained from tests made with fishes from lot number 1,the acetylrotenone and rotenolone curves from lot number 2 , and the rotenone hydrochloride curves from lot number 3. Lots 2 and 3 are considered identical since the differences fall within experimental error. 'The toxicity data are given in. Tables I1 to IV. The survival time curves and the velocity of fatality curves, which were plotted from these data, are given in Fig. 1 and 2. Comparative data obtained from the velocity of fatality curves are given in Table V. In each case the straight line which is an approximation of that portion of the curve corresponding to the greatest rate of increase in the velocity of fatality with increase in concentration is prolonged to cut the x-axis a t a point designated a; the slope of this line is designated tan 0.
W. A. GERSDORFF
1150
VOl. 55
TABLE 111 TOXICITY OF ACETYLROTENONE TO GOLDFISH AT 27.0 Mean length Mean weight of fishes, of fishes,' mm. g.
Concn., mg. per liter
No. of fishes used
0.50 .25 .15 .075 .050 .025
12 12 8 20 19 16 14
.020
8
37 (5)
.015
6
41
.010
7
b
.10
44 42 47 43 42 44 43
* 0.2"
Mean sum. time, min.
100
M~~~~ surv. time
2.6 98 2.3 96 3.1 120 2.4 180 2.3 229 2.6 362 2.4 Tenfishes,365'.
1.08 1.06 0.87 .60 .49 .31 Four .20" fishes still active after 48 hrs. 1.6 Five fishes, 904". Three .13' fishes still active after 30 hrs. 2.2 One small fish, 670". . 02" five fishes still active after 49 hrs.
*
Two small fishes affected a t first but apparently recovered. All active after 27 hrs.
a, b and c as in Table 11.
TABLE IV TOXICITY OF ROTENOLONB TO GOLDFISH AT 27.0 Concn., mg. per titer
No. of fishes used
3.0 2.0 1.5 1.0 0.75 .50 .40 .30 .25 .10
8 13 24
.050
Mean length Mean weight of fishes of fishes," mm. g.
41 43 42 43 44 45 42 40 41 3; (4)
23 18 7 13 7 15 6 10
f
0.2"
Mean SUN. time, min.
2.2 2.4 2.3 2.4 2.6 2.7 2.3 2.1 2.2 2.4
M~~~
118 122 136 143 178 199 250 358 448 856'
-~
100 surv. time
0.89 .84 .80 .72 .59 .52 .42 .28 .26 .12c
Apparently unaffected in 52 hrs.
a, b and c as in Table 11.
TABLE V
COMPARATIVE TOXICITY AT 27' AND
Substance
OF ROTENONE HYDROCHLORIDE, ACETYLROTENONE
ROTENOLONE TO GOLDFISH a', 'pg. per liter
Tan0 0 Minimum surv. liters per mg. per min. time, min.
