Vitamin B(B,) and G(3J Content of Cottonseed Products MAY L. WHITSITT,Southern Methodist University, Dallas, Tex.
C
OTTOK is a raw mateA comparative study of cottonseed meal, oil, and EVIDENCE OF THIRD FACTOR OF rial which is more and VITAMIN B COXPLEXIS hulls is made with respect to the vitamin B(B1) more closely allying the COTTOSSEED HULLS and eitamin G(B2) contents. The rat growth agricultural and chemical indusmethods are used. Cottonseed meal is a rich The average results showing tries. Because of the considerthe measure of vitamin B(B1) source of vitamin B ( B l ) and a good source of able i n d u s t r i a l importance of in cottonseed products are given cotton to the South and because eifamin G(BJ, though richer in the former than in Table I, and Figures 1 and 2. of the recognized n u t r i t i o n a l in the latter. Cottonseed oil contains no vitamin The data clearly show that this value of c o t t o n s e e d meal in nor d a m i n G(B2). The cottonseed hulls B(B1) sample of cottonseed meal is a cattle feeding, a study of cottonare as rich a source of z!itamin G(B2) as the rich source of vitamin B(Bl), seed products has been underthat it is richer than the same cottonseed meal. N o demonstrable amount of taken in order to learn more of weight of whole wheat ( 2 ) or their possibilities as nutrients. eifaniin B ( B J is found. skimmed milk powder (7), and, Stevens (8) in 1930 reported his T’I’hen an amount of cottonseed meal too small from r e s u l t s to be r e p o r t e d finding that cottonseedmeal was for complete protecfion against polyneuritis is later in this paper, that it is as a rich source of both vitamins f e d with the cottonseed hull extract and the citarich as the same weight of dry B(B), and G(B2). This invesbaker’s yeast. min B(BI)--deJicient diet, there is a much less tigation has for its object a comI n substituting the cottonseed parative study of c o t t o n s e e d rapid decline in weight and a much longer suroil for t h e b u t t e r fat in the meal, oil, and hulls with respect rival period than when the test animals receice basal diet, the vitamin A cont o their v i t a m i n B(B1) and the same amount of cottonseed meal without tent was decreased, but there vitamin G(B2) content. the hull exiracf. The animals of both groups was little reason to doubt that The rat growth methods for suffer polyneuritis. Since fhere is found no s u f f i c i e n t vitamin -4 for the d e t e r m i n a t i o n of v i t a m i n experimental period was B(B1) as developed by C h a s e appreciable amount of vitamin B ( B l ) in the hull supplied through storage in the and Sherman ( 2 ) and of vitaextract, these results are taken as ecidence fhut a n i m a l a n d c o d l i v e r oil in min G(B2) as d e v e l o p e d by the cottonseed hulls contain a third factor of the the diet. At no time was there Bourquin and Sherman (1) were oitamin B complex. any i n d i c a t i o n of vitamin A followed. The m a t e r i a l , the deficiencv. However. in order vitamin content of which was to be determined, was fed to the rats receiving diets deficient to have definite evidence that deficikncy of vitamin B(Bl) in vitamin B(B), or vitamin G(B2). The gron-th produced and not of vitamin A was responsible for the loss in weight in these rats in excess of that produced in the rats receiving in the animals, four rats were given the vitamin B(B1)only the diet deficient in vitamin B(Bl) or vitamin G(B2) deficient diet containing the cottonseed oil n-ith 2.4 grams is taken as a measure of the vitamin B(B1) or vitamin G(B2) a week of yeast powder. The animals ivithin 4 weeks had in the material. The conditions of the investigation were more than doubled their rreight. Since the yeast, according carefully controlled as to the care and selection of the animals to the reported assay and from the reports of many workers according to the above mentioned methods. All of the (4, 6 ) , contained no vitamin A, the large gains made by the experimental animals were bred from rats on a diet con- animals can be accounted for only through vitamin B(BI) sisting of two-thirds ground m-hole Theat, one-third whole of the yeast. From these results it is concluded that cottonmilk powder, and sodium chloride which was 2 per cent of the seed oil is as deficient in vitamin B(Bl) as butter fat, and it is weight of the wheat (6). suggested that cottonseed oil could be used instead of butter The cottonseed meal was fed separately from the basal as a source of fat in the vitamin B(BJ-deficient basal diet. diet in a weekly portion of 2.4 grams. The cottonseed RESULTSOF FEEDISG COTTOKSEED PRODoil was incorporated as the sole source of fat in the basal TABLEI. AVERAGE UCTS WITH VITAMINB(B,)--DEFICIENT DIET . , diets, thereby replacing the 8 per cent butter fat. The G % I NIN KEIGAT WEEKLY cottonseed hulls were extracted with purified 60 per cent AFTER: SCRVIYAL FOOD 5 weeks 8 weeks PERIOD INTAKE alcohol,’ and the extract was concentrated a t reduced tempera- RATS SUPPLEMEXT Grams Grams Dags Grams ture and pressure, and dried on cornstarch. This starch 10 -21.8 -23.4 29 15 0 15 2.4 grams meal a week 56 88 56 50 was incorporated in a food mixture otherwise like the basal 12 -21 16 Oil a t 8% -24 35 diet, replacing an equivalent weight of ordinary cornstarch. -27 10% hujiextract 12 -27 29 10 -32 -28 6 40% hull extract 29 10 The extract was fed a t 10, 20, and 40 per cent levels. In -21 0 6 gram meal a ueek 24 9 -24 37 4 0 % hull extract 4-0 6 one series the hulls were fed directly as hull bran in weekly 9 - 3 27 gram meal E u eek -21 54 portions of 2.4 grams. 1 T h e alcohol was purified a s follons, 10-20 grams potassium hydroxide were added t o a liter of 95 per cent alcohol T h e mixture stood seierai hours: 0 5 cc. saturated silver nitrate solution was slouly added T h e alcohol was filtered a n d distilled, t h e first a n d last 10-cc portions being discarded.
Eighteen rats received the vitamin B(B1)-deficient diet in which had been included the extract from the hulls.. Twelve of these animals received the extract from the hulls fed a t a 10 per cent level; six were fed the hull extract a t a 40 per cent
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level. By the fifth week only three animals from the total of both groups survived. All of them suffered from polyneuritis. The growth curve of these animals so nearly approximated that of the negative controls that no measura b l e a m o u n t of vitamin B(B1) could be considered present in the hull extract. A l t h o u g h it is known that in the cooking process of the cottonseed meal the gossypol seems t o be changed to a less toxic form (3), i t w a s d e c i d e d t o determine whether or not
with the diet containing the hull extract gave no significant difference from the average gain of animals due to the 2.4 grams of meal without the hull extract. However, the two groups receiving 0.6 gram of meal a week with and without hull extract gave very different results. Of the animals receiving 0.6 gram of meal with the vitamin B (BJ-deficient diet only, all but one had either died following development of severe polyneuritis or were suffering from the disease by the fifth week. One lived through 8 weeks ; polyneuritis developed the fifty-third day. The average loss in weight a t the end of the fifth week was 21 grams. The nine litter mates of these rats receiving 0.6 gram of meal a week and the vitamin B(B1)-deficient diet containing the hull extract showed a t the end of 5 weeks an average loss of only 3.3 grams. All but two of the animals survived the 8-week period but showed alternately the onset and partial or complete recovery of polyneuritis. At I the end of 8 weeks the average loss in weight was 21 grams. The average weekly food intake of the two groups showed no significant difference. 4 5-1 The hull extract seemed to be the cause of this less rapid z loss in weight, yet it apparently had nothing t o do with 5 2 the partial recovery from polyneuritis, as the diet containing U -3 the hull extract was always accessible t o the animal; but it was only after feeding the weekly portion of meal that there -4 was improvement in the conditions brought on by polyT I M E IN DAYS F I G U R E2. E F F E C TO F neuritis. The earlier results have shown that in the extract HULLS PLUS of hulls there is no vitamin B(B1); therefore, it seems that FIGURE 1. COMPARATIVE COTTONSEED AMOUNT OF COTTON- there must be some growth factor in the hull extract other EFFECTOF C O T T O N S E E D SMALL SEED MEAL ON RATS FED than vitamin B(B1) or G(B2). ON RATS FED A PRODUCTS B( B,)-DEFICIEKT VITAMIN B( BIFDEFICIENT A VITAMIN The animals receiving the larger quantity of cottonseed DIET DIET meal with or without the hull extract made such large and 1. Negative controls (10 rats) 1. Vitamin B(B1bdeficient diet 2. Cottonseed hull extract at 40 only (10 rats) rapid gains that probably they were not sensitive to the per cent level (18 rats) 2. Cottonseed hull extract at 40 effect of the hulls. 3. Cottonseed meal, 0.6 gram per per cent level (18 rats) VI
3.
4.
Cottonseed oil, 8 per cent (12 rats) Cottonseed meal, 2.4 grama per week (15 rats)
4.
a e e k (6 rats) Cottonseed meal, 0.6 gram per week, plus hull extract, 40 per cent (9 rats)
t h e rapid decline of the animals receiving the hull extract could be due to some toxic substance in the extract, although it was realized from the typical polyneuritic symptoms that insufficient amount of vitamin B(Bl) was an important factor in causing the decline. As the cottonseed meal, when it was fed with the vitamin B(B1)-deficient diet, had shown no evidence of any toxic material, it was decided t o feed the extract from the hulls with the cottonseed meal, together with the vitamin B(BJ-deficient diet. Four rats were given the vitamin B(B1)-deficient diet which had been made to contain hull extract a t a 10 per cent level. They also received 2.4 grams of cottonseed meal each week. There was no evidence of any toxic substance. But the four animals made a greater gain on the cottonseed meal fed with the hull extract than their litter mates made on the meal fed with the basal diet without the hull extract. Since the hulls had shown no trace of vitamin B(B1), it was thought the additional growth brought about by the meal plus the hull extract might be an indication of some groTYth factor in the hulls other than vitamins B(B1) and G(B2). To determine whether or not this indication was significant, a third series was started. With one group of nine animals the experiment was continued by feeding 2.4 grams of meal each week with the vitamin B (B1)-deficient diet containing 40 per cent hull extract. With nine litter mates of the above, the effect of feeding 0.6 gram cottonseed meal a week with the vitamin B(Bl)-deficient diet containing 40 per cent hull extract was determined. Six animals, litter mates of the above groups, were given the vitamin B(B,)-deficient diet plus 0.6 gram of cottonseed meal a week. The results of continuing the feeding of 2.4 grams of meal
EVIDESCE OF VITAMIX G(Bz) IN COTTONSEED HULLS The average results showing the measure of vitamin G(Bz) in cottonseed products are given in Table I1 and in Figures 3 and 4. Cottonseed meal is shown t o be a good source of vitamin G(B2), but it is richer in vitamin B(B1) than in vitamin G(B2). The cottonseed oil showed slightly less vitamin G(Bz) than butter fat. By means of autoclaved
FIGURE3. COMPARATIVE EFFECT OF COTTONSEED PRODUCTS ON RATS FED A \'ITAMIN G( B 2 ) - D ~ DIET 1. 2. 3
4.
Vitamin G(Bz)-deficient ( diet only (11 rats) Cottonseed oil (13 rats) Cottonseed meal, 2.4 grams per week 11.5 ata) (15 rrats) Cottonseed hulls, 2.4 grams Cottonseei per aeek (8 (a rats) ra~si
F I G U R E4. E F F E C T O F C O T T O N S E E DH U L L SO N RATSFEDA VITAMING( B2)~ ~ DEFICIEXT ~ ~ ~ ~DIET ~ 1.
2. 3.
4.
Vitamin G(B2)-deficient diet only (11 rats) Extract of hulls fed at 10 per cent level (16 rats) Extract of hulls fed at 20 per cent level (8 rats) Hulla unextracted fed at 5 t o 7 per cent level (8 rats)
yeast it was established that the decline in weight of the animals receiving the basal diet containing cottonseed oil was not due to vitamin A deficiency. This oil could be conveniently substituted as a source of fat for the butter in the vitamin G(B2)-deficient basal diet. The results of feeding the hull extract a t a 10 per cent level gave evidence of some vitamin G(B2), and because of
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IXDUSTRIAL AND EKGINEERING CHEMISTRY
this evidence a further investigation of the hulls was undertaken. I n the second series the hull extract was incorporated with the vitamin G(B2) basal diet a t a 20 per cent level. Eight rats were given this diet. To eight litter mates of these, carefully matched as to weight and sex, were given 2.4 AVERaGE RESULTS O F FEEDING COTTONSEED PRODCCTS WITH VITAMING ( B 2 ) - - D ~ ~ 1 cDIET 1~x~
TABLE11.
R.ATS 7 13 15 16 8 8
GAININ WEIGHTAPTER: 5 weeks 8 weeke Grams Grams 0 3 3 8% cottonseed oil -3 -6 2.4 grams cottonseed meal per week 15 2s 2 7 10% hull extract 20% hull extract 11 17 2.4 grams hiill bran per week 23 32 SUPPLEMEKT
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per cent level) made an average gain of 10 grams. The second group (receiving hull bran) a t 5 weeks had made an average gain of 23 grams; at' the end of 8 weeks the gain was 32 grams. The cottonseed hulls are as good a source of vitamin G(B2)as the cottonseed meal. LITERATURE CITED
(1) Bourquin, A , , and Sherman, H. C., J . Am. Chem. Soc., 53, \?OF EAHHLEy FOOD 3501-6 (1931). PER R . ~ T (2) Chase, E., and Sherman, H. C . , I b i d . , 53, 3506-10 (1931). Grams (3) Clark, E. P., J . Bid. Chem., 78, 159-66 (1928). 26 (4) Honeywell, H., and Dutcher, R. E., J . Sutrition, 3 , 4 9 (1931). (5) Osborne, T. B., and Mendel, L. B., J . Biol. Chem., 45, 277-88 (1921). 23 22 (6) Sherman, H. C., and Campbell, H. L., I b i d . , 60, 5-15 (1924). 30 (7) Sherman, H. C., and Whitsitt, M. L., Ibid., 90, 153-60 (1931). (8) Stevens, Henry, Oil & Fat I n d . , 7, 215-16 (1930).
gi
grams a week the 'Ulextracted which amount Was b e t w e n 5 and 6 per cent of the food intake. At the end of 5 m-eeks the first group (receiving the hull extract a t 20
RECEIVED .Ipril 20, 1933. Presented before t h e Division of Agricultural a n d Food Chemistrv a t t h e 85th Meeting of t h e American Chemical Society, Washington, D C ,-March 26 t o 31,1933.
Shortening Value of Plastic Fats JENNIE D. FISHER,Institute of American Meat Packers, Chicago, Ill.
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point, and plasticity as factors. 0 D E R K technology A standard procedure has been worked out in L a n g m u i r (5) d i s c u s s e s the has made available a determining the relative shortening value of f a t s greater covering power of unwide a s s o r t m e n t of by means of the Shortometer such that the resaturated fatty acids, a property e x c e l l e n t cooking fats, each sulting data meet the requirements of statistical significant in the present conwith characteristic merits. The measuremenis as to scope and validity of the nection. Harkins (4)speaks of discriminating h o u s e w i f e o r the adhesive properties imparted baker will increasingly s e l e c t mean. by double b o n d s , thus giving fats with a view to adapting the The large range of excellent cooking f a t s now insulating strength to the layers special merits of each to the reavailable makes desirable a revaluation of their of fat separating the gluten. I n sult sought. This calls for a qualities for the rarious purposes of cookery. the present study the only conrevaluation for cookery purposes The present study was directed at one of these stant among those determined on of available, edible fats, espethe fats used which coordinated cially since much of the present qualities-i. e., shortening value in plain p i e with shortening value was, posliterature of the subject dates crust. CTse of 41 and 44 per cent of f a t , Jive sibly, the c o n g e a l i n g p o i n t . from the war period a t which lards, two hydrogenated cottonseed oils, a hydroShortening value in the present time emergency considerations genated lard, a n animal stearin-vegetable oil prevailed in such investigations. state of our knowledge must recompound, and an all vegetable oil compound, main a matter of direct test. The present study had for its purpose the determination of the The present study sought to showed shortening ealues as determined by the relative value of some common a p p r o a c h t h e problem f r o m Bailey Shorlometer which coordinated approxifats for a particular use-i. e., as the standpoint of ample statismately with their congealing points. FVhen a shortening in the making of tical scope a n d c r i t e r i a . A amounts of f a t were used i n fhe formula in the plain pastry or pie crust. While thousand pie crust wafers were inverse order of their shortening z'alue, the the action of a shortening agent b r o k e n for each fat, and the is understood, the relation bevalidity of the average of these breaking strengths came out approximately the tween the shortening power and tests was d e t e r m i n e d by the same. the inherent properties of a fat application of the usual measis not understood. Shortness is urements such as the frequency the quality of cookies, crackers, and pie crust which makes distribution, median, mode, arithmetic and geometric mean, them tender and easy to break or crush. A mixture of flour, average and standard deviation, probable error, and Pearson's water, and salt bakes to a hard mass, owing to the adherence coefficient of variation. Fisher's "t'was used to test the and continuity of the gluten in the flour. When a fat is significance of the difference between any pair of means being added, being immiscible JTith the other constituents, it compared. spreads and separates the gluten particles. The resulting TESTSOF SHORTESISG\'ALCE shortness is measured in a delicate breaking machine, the Shortometer, first introduced by Davis (1-3), and since SERIESI. The following fats were used in the first experiimproved, especially by H. C. Bailey. ment: a refined lard, a partially hydrogenated lard, an With reference to the relation between the action of a animal stearin-vegetable oil compound, and two partially shortening agent and its inherent properties, Platt and hydrogenated cottonseed oils. Fleming (6) enumerate viscosity and surface tension, melting Sufficient materials to carry out a complete set of tests for I
