Summary In a series of four tests in which three deliberately chosen toxic cottonseed meals were treated with aqueous solutions of salts and alkalies, it was found that the best detoxifying effect was obtained with sodium hydroxide, followed very closely by potassium and ammonium hydroxides. Dry heat treatment alone did not detoxify, and mortality was high. Treatment with moisture plus heat gave partial detoxification. Of the 22 chemically‐treated cottonseed meal samples tested, those treated with alkalies showed the best weight gains, the order of decreasing effectiveness being NaOH>KOH>NH4OH>italicCafalse(italicOHfalse)2. The residual toxicity of treated cottonseed meals cannot be explained on the basis of their free gossypol content as analyzed for meals with high values gave better growth performance than some with lower levels of free gossypol. There were also very marked differences in final body weight after 8 weeks of feeding six different treated cottonseed meal samples having practically the same free gossypol content.
I n a recent paper Castillon and Altschul ( 2 ) described experiments on the toxicity of some new water-soluble combination products of gossypol to aquarium fish. The present investigation deals with the acute oral toxicity and effects of the body weight of the rat of identical and similar combinations of gossypol with glycine, dextrose, and peanut protein. The results of incorporating in the diet various levels of pure gossypol and cottonseed pigment glands are also reported. An understanding of the physiological effects bears direct application to the improvement of the food of man and animals. METHODSSamples of untreated cottonseed pigment glands, pure gossypol, and combination products with gossypol were prepared as described (8,5 ) . Individually caged male rats (150-220 g.) were fasted for 18 hours, with water ad libitum, in an air-conditioned animal room maintained a t 80" 2 1°F. and ca. 45% relative humidity. Single doses of the test materials were administered by stomach tube as 10% solutions or suspensions in distilled water. All rats had free access to stock diet and water after intubation. The per cent composition of the control and basal ration was: whole ground wheat 21.0, meat and bone scraps 18.0, skim milk 15.0, soybean oil meal 15.0, yellow corn 13.0, lard 10.0, yeast and salt mixture 2.5, liver and glandular meal 2.0, alfalfa leaf meal 2.0, wheat germ oil 1.0, vitamin A and II oil (2250 U.S.P. A, 300 U.S.P. Ds per g.) 0.5. The surviving rats were weighed daily or 3 times per week for periods of 2 weeks or more. Calculation of the LDso value was made after 1 week by the method of Reed and Muench (7). All gossypol analyses reported in this study were made a t the Southern Regional Laboratory by the method of Pons and Guthrie (6). I n many cases 4 or more individual analyses were made and the average of these is shown as the gossypol content.I n the 4 subacute toxicity experiments, levels of 0, 0.05, 0.075, 0.10, 0.15, 0.20, 0.30, 0.40, 0.50, 0.536, and 1.34 g. of pure gossypol and levels of 1.34 g. of cottonseed pigment glands containing 21.85% gossypol and 0.6 and 0.9 g. of cottonseed pigment glands containing 33.22% gossypol were added per 100 g. of stork ration and fed ad Zibtfum to weanling or young adult male rats. These rats were weighed daily for the first 14 days, and at least thrice weekly thereafter. RESULTSThe samples of gossypol, untreated cottonseed pigment glands, and gossypol combination products are listed in Table 1 in the order of increasing acute oral toxicity. The LD,, value recorded for pure gossypol administered as an aqueous suspension was not significantly different from the LD,, value for pure gossypol dissolved in soybean oil. There was poor correlation between the gossypol content of the various samples and their LD,, values. For example, Sample F, which contained 68.6% extractable gossypol, was more toxic than 100% gossypol, and Sample D, containing 543 TABLE 1
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