A134-231-6000-L180 TEXAS AGRICULTURAL EXPERIMENT STATION A. B. CONNER, DIRECTOR COLLEGE STATION, BRAZOS COUNTY, TEXAS BULLETIN NO. 422 MARCH, 1931 DIVISION OF CHEMISTRY Variations in Vitamin A and Chemical Composition of Corn AGRICULTURAL AND MECHANICAL COLLEGE OF TEXAS T. O. WALTON, President STATION STAFFT ADMINISTRATION: A. B. CoNNER, M. S., Director R. E. KARPER, M. S., Vice-Director CLARIcE MIxsoN, B. A. Secretary M. P. HOLLEMAN, JR., Chief Clerk J. K. FRANcKLow, Assistant Chief Clerk CHEsTER HIGGS, Executive Assistant C. B. NEBLETTE, Technical Assistant CHEMISTRY: G. S. FRAPs, Ph. D., Chief; State Chemist S. E. AsRuRY, M S., Chemist J. F. FUDGE, Ph. D., Chemist E. C. CARLYLE, B. S., Assistant Chemist WALDo H. WALKER, Assistant Chemist VELMA GRAHAM, Assistant Chemist T. L. OGIER, B. S., Assistant Chemist ATHAN J. STERGES, B. S., Assistant Chemist JEANNE M. FuEoAs, Assistant Chemist _ RAY TREIcHLER, M. S., Assistant Chemist RALPH L. ScHwARTz, B. S., Assistant Chemist C. M. PQUNDERs, B. S., Assistant Chemist HORTICULTURE: ' S. H. YARNELL, Sc. D., Chief L. R HAWTHORN, M. S., Horticulturist RANGE ANIMAL HUSBANDRY: J. M. JoNEs, A. M., Chief B. L. WARwIcK, Ph. D ., Breeding Investigations STANLEY P. DAvIs, Wool Grader ENTOMOLOGY: F. L. THOMAS. Ph. D., Chief; State Entomologist . J. REINHARD, B. S., Entomologist . K. FLETcHER, Ph. D., Entomologist . L. OWEN, JR., M. S., Entomologist . . RONEY, M. S., Entomologist _ . . GAINEs, JR., M. S., Entomologist . . JoNEs, M. S., Entomologist . BIBBY, B. S., Entomologist IL E. HEARD, B. S., Chief Inspector o MAcKENsEN, B. S. Foulbrood Inspector . B. WHITNEY, Foulbrood Inspector (nLd-iéwm "$1102 > o m g gévgm z o 2 f.‘ . B. REYNQLDS, Ph. D., Chief ‘ SUBSTATIONS R. E. KARPER, M. S., Agronomist P. C. MANcELsDoRF, Sc. D., Agronomist D. T. KILLOUGH, M. S., Agronomist H. E. REA, B. S., Agronomist , Agronomist B. C. LANGLEY, B. S., Assistant in Soils No. 1, Beeville. Bee County: ¢ R. A. HALL, B. S., Superintendent No. 2, Troup, Smith County: _ P. R. JOHNSON, M. S., Superintendent No. 3, Angleton, Brazoria County: R. H. STANSEL, M. S., Superintendent No. 4, Beaumont. Jefferson County: R. H. WYcHE, B. S., Superintendent No. 5, Temple, Bell County: _ HENRY DUNLAvY, M. S., Superintendent B. F. DANA, M. S., Plant Pathologist H. E. REA, B._S., Agronomist; Cotton Root Rot Investigations _ SIMON E. WoLI-"F, M. S., Botanist; Cotton Root Rot Investigations r No. 6, Denton, Denton County: P. B. DUNKLE, B. S., Superintendent No. 7. Spur, Dickens County: _ R. E. DICKSON, B. S., Superintendent i-———i, Agronomist No. 8, Lubbock, Lubbock County: D. L. JoNEs, Superintendent FRANK GAINEs, Irrigationist and Forest Nurseryman No. 9, Balmorhea, Reeves County: J. J. BAYLEs, B. S., Superintendent Teachers in the School of Agriculture Carrying Cooperative Projects on the Station: W. ADRIANcE, Ph. D., Horticulture W. BILSING, Ph. D., Entomology P. LEE, Ph. D., Marketing and Finance SCOATES, A. E., Agricultural Engineering . K. IVIACKEY, M. S., Animal Husbandry *Dean School of Veterinary Medicine. **In cooperation with U. S. Department of Agriculture. q _ POULTRY HUSBANDRY: PUBLICATIONS: . D. JACKSON, Chief VETERINARY SCIENCE: *M. FRANcis, D. V. M., Chief H. ScHMIDT, D. V. M., Veterinarian '_ F. P. MATHEws, D. V. M., M. S., Veterinari . T. HARDY, D. V. M., Veterinarian ‘ . E. CARRoLL, D. V. M., Veterinarian as ‘U P‘ > Z -1 w: i» a m o P‘ o Q -< ='> z U w I -< (D h! o P‘ o o r4 n J. TAUBENHAUS, P . D., e . N. EZEKIEL, Ph. D., Plant Pathologist J. BAcH, M. S.. Plant Pathologist -—-—-'—-—.i, Plant Pathologist M AND RANCH ECONOMICS: . P. GARBARD, ., Chi . E. PAuLsoN, Ph. D., Marketing . A. BoNNEN, M. S., Farm Management . R. NIsBET, B. S.. Ranch Management . C. MAGEE, M. S., Farm Management RURAL HOME RESEARCH: ' JEssIE WHiTAcRE, Ph. D., Chief MARY ANNA GRIMEs, M. S., Textiles ELIZABETH D. TERRILL, M. A., Nutrition SOIL SURVEY: **W. T. CARTER, B. S., Chief E. H. TEMPLIN, B. S., Soil Surveyor A. H. BEAN, B. S., Soil Surveyor R. M. MARSHALL, B. S., Soil Surveyor BOTANY: V. L. C0RY, M. S., Act. Chief SIMoN E. WOLFF, M. S., Botanist SWINE HUSBANDRY: FRED HALE, M. S., Chief DAIRY HUSBANDRY: . A O. C. CoPELAND, M. S., Dairy Husbandman ' Esnsri es“ R. M. SHERWOOD, M. S., Chief AGRICULTURAL ENGINEERING: H. P. SMITH, M. S., Chief MAIN STATION FARM: G. T. McNEss, Superintendent APICULTURE (San Antonio): H. B. PARKS, B. S., Chief A. H. ALEX, B. S., Queen Breeder FEED CONTROL SERVICE: F. D.‘ FULLER, M. S., Chief S. D. PEARcE, Secretary J. H. RocERs, Feed Inspector K. L. KIRKLAND, B. S., Feed Inspector SIDNEY D. REYNQLDs, JR., Feed Inspector P. A. M00RE, Feed Inspector E. J. WiLsoN.,B. S.. Feed Inspector H. G. WICKES, B. S., Feed Inspector No. 10, College Station, Brazos County: R. M. SHERWOOD, M. S., In charge ' L. J. McCALL, Farm Superintendent No. 11, Nacogdoches, Nacogdoches County: H. F. MoRRIs, M. S., Superintendent ’ **No. 12, Chillicothe, Hardeman County: J. R. QUINBY, B. S., Superintendent _ **J. C. STEPHENS, M. A., Assistant Agroncm No. 14, Sonora, Sutton-Edwards Counties: W. H. DAMERON, B. S., Superintendent -———-i———, Veterinarian W. T. HARDY, D. V. M., Veterinarian **O. G. BABCOCK, B. S., Entomologist O. L. CARPENTER, Shepherd N0. 15, Weslaco, Hidalgo County W. H. FRIEND, B. S., Superintendent SHERMAN W. CLARK, B. S., Entomologist W. J.-BACH, M. S., Plant Pathologist No. 16, Iowa Park, Wichita County: N C.1H. McDowELL, B. S., Superintendent o. 1, --—i-€-— N0. 18, ——— —-——?————-, Superintendent No. 19, Winterhaven, Dimmit County: E. MoRrENsEN, B. S., Superintendent N L2H. HAWTHORN, M. S. , Horticulturist o. 0, €—-——-- , Superintendent , Superintendent J. S. IVIOGFORD, M. S., Agronomy F. R. BRIsoN, B. S., Horticulture W. R. HoRLAcHER, Ph. D., Genetics J. H. KNox, M. S., Animal Husbandry TAs of March 1, 1931. Deficiency of vitamins, protein, energy, or minerals, in the diet of man or animals may result in decreased health and vigor or in certain diseases. Vitamins are organic compounds found in foods in very small quantities, and are essential to the life and health of animals. Albino rats are used to esti- mate the quantity of vitamins in foods. Methods are de- scribed for breeding the rats and testing the foods for vitamin A. Selection of the breeding rats resulted in improvement in uniformity. The vitamin A content of yellow corn is high. One gram of yellow corn (39 samples) contained 2.5 to 8 units of vitamin A, one gram of red or variegated corn (18 samples) contained 0.9 to 5 units, while one gram of white corn usually contained less than .03 unit. The units of vitamin A in crosses of yellow with white corn were approximately in proportion to the num- ber of genetic factors for yellow color in th_e crosses. Season and locality (11 localities and 3 seasons) had some elfect on the vitamin A content of yellow and of red corn. Varieties of corn varied little in composition, including protein, but the protein content varied considerably according to the locality in which the corn was grown. The correlation between rainfall, January through July, and protein content was —.576i.072. Slightly less lime and phosphoric acid were in the corn grown in two localities than in the samples grown at the other five places. TABLE OF CONTENTS Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Vitamins and their importance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ' ‘Vitamin A in corn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. p Methods of estimating vitamin A . . . . . . . . . . .1 . . . . . . . . . . . . . . . .. ~7~ Standardization and care of rats . . . . . . . . . . . . . . . . . . . . . . .. 81 Details of the ration method for estimating vitamin A . . . . . . . . 10- Results with the ration method . . . . . . . . . . . . . . . . . . . . . . . . . .. 12 The unit method.‘ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 12: Comparison of the unit method and the ration method . . . . . .. 20 Modification for corn low in vitamin A . . . . . . . . . . . . . . . . . . .. 2O Results of tests with the unit method. .4 . . . . . . . . . . . . . . . . . . . . . . .. 21 * Vitamin A in yellow corn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 21; Vitamin A in red and in Variegated corn . . . . . . . . . . . . . . . . . . . 291 Vitamin A in white corn. i . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31A Relation to heredity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 31 Variations according to season and locality. . . .i . . . . . . . . . . . . . . . .. 32 Corn meal and hominy feed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 34‘ Composition of corn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 345p Variation in protein . . . . . . . . . . . . . ». . . . . . . . . . . . . . . . . . . . . . . . 35 1 Variation in minerals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 35 Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 44 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 PAG i‘ BULLETIN NO. 422 _ MARCH, 1931 VARIATIONS IN VITAMIN A AND IN CHEMICAL COMPOSITION OF CORN G. S. FRAPS Sufficient energy, proteins, vitamins, and minerals, such as phosphoric acid, lime, magnesia, sodium, chlorine, sulphur, potash, iron, copper, iodine, and fluorine must be furnished by the diet of man and animals, according to recent investigations. Deficiencies in diet may cause re- tardation in growth of young animals, deficiency of production such as milk, or eggs, lack of vitality or vigor; or may lead to diseases such as xerophthalmia, pellagra, scurvy, anemia, rickets, and goiter, as well as susceptibility to other diseases such as pneumonia or tuberculosis, or to decreased ability to reproduce. - Vitamins are organic substances which are present in very small quantities in foods, and are known to be essential to the health of ani- mals. Vitamins vary in amounts in different foods. A particular food may contain a large amount of one vitamin, while the amount of one or more of the others may be very low, or it may be almost entirely absent. A deficiency of any vitamin in the diet may result in failure of young animals to grow, disturbances of bodily function, disturbances of health, or even in serious diseases such as rickets, scurvy, or beri-beri. Vitamin A was one of the first vitamins discovered. It occurs in large quantity in yellow corn, while little or none is found in white corn. For the purpose of this study, samples of corn grown at the various substations in different parts of the state were furnished by the Division of Agronomy. Chemical analyses were made in addition to the estima- tion of vitamin A ; and as the analyses bring out some points of interest, they also are discussed in this Bulletin. VITAMINS AND THEIR IMPORTANCE As previously stated, vitamins are organic substances found in small amounts in foods, and are essential to the health or proper functioning of the animal body. The exact number of vitamins is not known, since new ones are being discovered from time to time and those now consid- ered as single vitamins may later be split into two or more. Vitamins are studied by means of experiments on animals; and complete or par- tial lack of one of them is recognized by the failure of the animal body to perform some of its functions. The following is a brief description of the vitamins (19) known at present. Vitamin A- This is called also fat-soluble A, or the anti-xerophthalmic vitamin. It is essential for bodily health-and growth. An eye trouble now believed to comprise at least three vitamins of which two have =1’, 6 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION called xerophthalmia frequently occurs when vitamin A is not consuo in sufficient amounts. Young‘ animals do not grow well when the d' is deficient in vitamin A, and also are likely to suffer from the sore e referred to above, or from other disorders. Vitamin A is abundant‘ such foods as green vegetables, milk, butter, and yellow corn. i amount is low in rice, Wheat flour, white corn, oats, and other foods. g Vitamin B COInDIBX- What was called Vitamin B a short time ago; been given three different names. (B, B1 or F; B2, G or P-P; B3 . possibly B4.) So much work has been done and apparently is a being done on the complex termed vitamin B, that it would introd -_ much confusion to retain the term vitamin B for one of the constituent " of the vitamin B complex. In accordance with the proposal of S." _ Smith (18, see also 3), the two chief constituents of the vitamin: complex are here termed vitamin F and vitamin G. Vitamin C- Vitamin C is also termed the anti-scorbutic vitam Deficiency of vitamin C results in a disease known as scurvy. T disease was formerly of frequent occurrence in time of famine or long sea voyages and it occurred also during the world war. For a hundred years it has been known that lemons or vegetables wo prevent scurvy. Vitamin C occurs in large quantities in tomatoes, w? cabbage, turnips, and citrus fruits, especially lemons and grapefr y Wheat flour, corn meal, oat meal and similar cereals contain lit‘ vitamin C. It may be destroyed by cooking. * Vitamin D- This is also called the anti-rachitic vitamin. Deficien of vitamin D results in imperfect assimilation of lime in the bon, causing young animals or children to suffer from rickets, one sympto, of which is soft bones. A deficiency of vitamin D may also be co, ducive to decay of the teeth. Vitamin D occurs in liver, egg yolk, y] in small amounts in milk and butter. The compound formed by e posure of oils or certain vegetable waxes to ultra-violet light rays a in the same way as vitamin D. Sunshine takes the place of vitamin i‘. Animals or people who get plenty of sunshine should not suffer i. deficiency of vitamin D. With the abundance of sunshine available Texas, there is little reason for children or animals to suffer fro rickets, although it does occur. Cod liver oil is rich in vitamin D. i A Vitamin E- This is the reproductive vitamin, and is needed for reproduction of animals. Deficiency of this vitamin results in sterili Vitamin E is widely distributed, being present in wheat, oats, m' » certain vegetable oils, lettuce, and other foods. Vitamin F - Vitamin F is the anti-neuritic vitamin, soluble in Wat’ and destroyed partly or completely by heat (1). Its presence is j quired to prevent polyneuritis in birds and animals,‘ and possibly be beri in man. This vitamin has also been termed vitamin B1, or. vita" VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 7 B. Complete absence of vitamin F is followed by loss of appetite, loss of weight, and a condition of nervousness. It is found in wheat, corn, yeast, rice polish, and in other foods. Vitamin G- (5) This is known as the anti-pellagric vitamin and is a water-soluble vitamin, not easily destroyed by heating. IVhen the ration of young rats is dificient in this vitamin, there occurs a rapid decrease in rate of growth. Older animals may lose weight. The rats also become weak and may develop skin lesions, which are symptoms like those which occur in pellagra. Cereal grains are poor in vitamin G, while milk and green leafy vegetables seem to be better supplied with it. Vitamin Ba- (4) This is a water-soluble vitamin, recently discovered in the vitamin B complex. It is destroyed by heat. It is necessary for the growth or recovery in weight of birds, but does not seem to be needed by rats. Yeast, whole grains, and malt are good sources of vitamin B3; beef and beef liver are fair sources; milk, orange and tomato juice, spinach, potato juice, and corn molasses are somewhat low in this vitamin, ’ Vitamin 13,- (11)' This has been reported as present in the vitamin B complex but little definite information is available regarding it. It is similar to B3 but is needed by rats, while B3 is needed by birds but not by rats. VITAMIN A IN CORN Steenbock in 1919 (14) first showed that yellow corn contains a much higher content of vitamin A than does white corn. Steenbock and Coward in 1927 (15) found corn deficient in vitamin D. Bocke and Hetler in 1928 (10) studied the vitamin A in corn and its distribu- tion in the by-products from the manufacture of starch. The greater part of vitamin A was located in the pigmented layer of the endosperm. Corn bran and grits were deficient in vitamin A. Only a few quantitative determinations of the content of vitamin A in corn have been made. M. C. Smith (16, 1'7) in 1930 found that 0.3 gram of yellow corn contains one unit of vitamin A compared with about 5.5 gm. for hegari and 3 gm. for yellow milo. Russell (11) found about 0.5 gram of yellow dent corn contained one unit of vitamin A, while white-topped yellow dent contained one unit in 0.75 gm. Hauge and Trost (61) compared the growth of rats on five lots of corn of different breeding and concluded that the vitamin A was transmitted exclusively‘ with the genetic factors for yellow endosperm. Meyer and Hetler (4) studied the vitamin A concentration in the by-products from the manufacture of starch. METHODS OF ESTIMATING VITAMIN A While colorimetric tests have been proposed for the estimation of vitamin A, they cannot be considered reliable. It is necessary to use O 8 BULLETIN N10. 422, TEXAS AGRICULTURAL EXPERIMENT STATION biological methods for the detection and estimation of vitamins. Ra rabbits, guinea pigs, pigeons, chickens, and other animals are» used. Two methods were used for the estimation of vitamin A in the wo reported here. In both methods, the amount of vitamin A was ured by the growth of rats. One method, here termed the rati method, consists of feeding the animal an unlimited quantity of ‘A ration presumably sufficient in all nutritive factors except vitamin i? in which vitamin A is supplied only by the food to be tested. This ' the older method. The other method, here termed the unit meth_ (12,155), consists in feeding rats previously freed of vitamin A, 1 weighed amount of the feed to be tested, in connection with a ratio complete except for vitamin A. Both methods depend upon the ga” in weight of the rats, and as the growth of rats is subject to oth factors such as capacity to grow and infections or other diseases, if methods do not possess the exactness of chemical analysis. Standardization and Care of Rats The rats used were progeny of Albino rats obtained from the Wis = Institute, and from the Albino- Supply Company, Philadelphia. Th rats were paired and kept in cages of wire cloth, 12 inches wide, 1, inches long, and nine inches high. These cages were placed over pa on wooden stands constructed in such a way as to hold sixteen cages ' four tiers, an arrangement that allows the cages to be taken do l] readily for cleaning. An automatic watering device kept the rats su‘ plied with water, and a satisfactory ration was kept before them. Piec’ of bone were given them on which they could grind their teeth. Pot toes, lettuce, or canned spinach were given once a week. The ratio which was found to be satisfactory, was as follows: ' Corn meal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..1200 grams Cottonseed meal . . . . . . . . . . . . . . . . . . . . . . . . .. 200 grams‘ Powdered whole milk . . . . . . . . . . . . . . . . . . . .. 600 grams Alfalfa meal . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 40 grams Sodium chloride . . . . . . . . . . . . . . . . . . . . . . . . .. 10 grams Calcium carbonate . . . . . . . . . . . . . . . . . . . . . . .. 10 grams An analysis of one sample of this mixture follows: Protein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17.14 Fat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 8.17 Crude Fiber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.45 Nitrogen-free Extract . . . . . . . . . . . . . . . . . . . . . . . . . . .60.14 Moisture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..~. 8.86 Ash . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2.45 The system of selecting and keeping up the stock of breeding ra :r was as follows: a Ten pairs of rats which came as nearly as possible within the follow4 ing requirements were selected and mated each month: “ VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 9 (a) The parent should have raised two litters in four months. y (b) Sixty per cent of litters should have been suitable for experi- .-, mental work. (c) The parents should not have produced more than three litters when the young were selected for breeding. (d) The selected rats should be healthy. Thirty young rats were chosen carefully each month, weighing 35 to 45 grams when 21 to 28 days of age. Females and males were kept separately until grown, in large cages 24 inches by 18 inches by 9 inches, provided with circular revolving ' discs for exercising. 1 At the age of 120 days, suitable rats weighing 160 to 180 grams in ‘the case of females and 220 to 250 gramsin the case of males, were Qpaired in regular breeding cages. A Breeding rats that failed to produce offspring for a period of 3 months were destroyed. e I In this way the stock has been successfully replenished and a con- tinuous supply of young rats made available, the larger proportion of which falls within the weight required for the experimental work. The ‘system of selection described was more successful than anticipated. At the beginning of the work, 40 to 50 per cent of the young rats were too mall or too large for the requirements of the work. By the end of ‘ g " years, practically all the young rats could be used in the experiments. q , In the fall of 1929, the pair system of breeding described above was mpared with the group system used at the Pennsylvania Station. In e group system, one male and four females are placed in large breeding ges. The rats are numbered by marks on the ears. Pregnant females re isolated in individual cages. After the young are weaned, the emale is placed with other females for a resting period of 14 days fore being returned to the cage containing the same male to which 1e was previously mated. -_ The results of comparison of the two methods for four months are 'ven in Table 1. While more rats were born by the pair method than y i the group method, a smaller number was raised. The superiority of a ‘=4 group method was probably due to the period of isolation from the ale and the rest given the female. The new system also had the ad- Wntage of requiring a smaller number of males and a smaller number cages. The entire colony was placed on the group system in March, 930. Table 1.—Number of rats by the paired method and by the group method. Paired Group method method mber of females . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ; . . . . . . . . . . . . . . . . 45 46 I ber 0f young rats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 371 313 107 188 lmber of rats raised . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v entage raised . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 60 erage number of rats raised per female . . . . . . . . . . . . . . . . . . . . . . . . 2.4 4.1 10 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Details of the Ration Method for Estimating Vitamin A In the first series of tests, the method here called the ration method was used for comparing the amounts of vitamin A in the differenl samples. In this method, the young rats were fed a mixture comple i as nearly as possible in all respects except in vitamin A. It shoul contain the vitamin A only in the sample to be tested‘. The rats wer allowed "to eat freely of the ration, and the amount of vitamin A measured by the gain in weight over a period of 8 weeks or more. ' At first the immediate objective was to secure a ration that woul give normal growth over an experimental period of approximately thr months. With this object in view, response in growth to varyin amounts of protein, carbohydrate, fat, salts, and yeast was determine Casein proved a satisfactory protein but unfortunately the crude case' contains appreciable traces of the vitamins associated with milk and was necessary, therefore, to purify it by extraction with a suitable solven. or otherwise. It was boiled with a mixture of equal parts of ether ri» alcohol for one hour under a reflux condenser; after three successiv extractions the extracted casein was filtered off by suction and dried a 100 degrees C. A The requisite minerals were supplied by using a modification of th salt mixture of Osborne and Mendel, which was prepared as follows: Salt Mixture '7 015 Dicalcium phosphate . . . . . . . . . . . . . . . . . . . . . . . . . .180 gm. 1; Potassium sulphate . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1'7 gm. 3 Potassium chloride . . . . . . . . . . . . . . . . . . . . . . . . . . . .111 gm. Calcium carbonate . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 gm. , Magnesium carbonate . . . . . . . . . . . . . . . . . . . . . . . . . 24 gm. I Sodium carbonate . . . . . . . . . . . . . . . . . . . . . . . . . ..' . . 34 gm. Potassium carbonate . . . . . . . . . . . . . . . . . . . . . . . . . . 35 gm. Citric acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .111 gm. Iron, citrate (or ferrous sulphate 2 gm.) . . . . . . . . . . 6 gm. ,1; Potassium iodide (1 gm. in 500 "cc., take 10 cc.) . . . . .02 gm. Manganese sulphate (4 gm. in 500 cc., take 10 cc.) .08 gm. Sodium fluoride (12.5 gm. in 500 cc., take 10 cc.) . . .25 gm. Potash alum (1 gm. in 500 cc., take 10 cc.) . . . . . . . .02 gm. All the salts were finely powdered, and then mixed well. The sol}; tions to be used were placed in an evaporating dish; 100 cc. wa A. added, and mixed well with the salts; the mixture was dried in a stea bath, and then powdered. . . ._ Fleischmann’s stock yeast was found to give satisfactory results asp i" source of vitamin B when 10 per cent was used. With smaller poi ' centages the rats began losing weight at the end of 5 or 6 weeks inste": of gaining. ' VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 11 The following ration was used in the experiments (Mixture 639) : Salt mixture (Method 7015) . . . . . . . . . . . . . . . . . . .. 8 parts Casein (purified) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 32 parts Yeast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 1O parts Wesson oil (cottonseed oil) . . . . . . . . . . . . . . . . . . . . .. 20 parts Corn to be tested . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .130 parts The technique employed was as follows: Four or more young rats 23 to 26 days old and Weighing 38 to 42 grams, were placed in circular experimental cages (tinned test-tube baskets) standing on 10-inch soup plates, two of the same sex being placed in each cage. The rats did p not have access to their excretions, which contain vitamin A. Samples ' of corn were fed freely in Mixture 639 in granite-ware cups, so that the rats had all they could eat. The Weights of the rats and of the food eaten were recorded each Week. A number of checks Were run in the course of the work, by substituting 2 per cent of cod liver oil for 2 per cent of Wesson oil in the ration mentioned above (No. 639), both with starch and with several of the different varieties of corn. The checks gave satisfactory results, as the V‘ rats grew well. Origin 0f C0111 Used- The samples of corn Were secured from the various experimental farms through the Division of Agronomy. The soil type, on which the corn was grown, year of collection, and fertilizer used are given below. Beaumont, No. 4—Lake Charles clay loam and Crowley clay, 1926-7-8-9, no fertilizer. N acogdoches, No. 11—Kirvin clay loam, 1926. Kirvin fine sandy loam, 1927, 1928, 1929. Fertilized in 1926; fer- tilized with 300 pounds of 4-12-2 in 1927, 1928; 400 pounds of 6-9-3 in 1929. Angleton", No. 3 Lake Charles clay loam, 1926, 1927, 1928. Lake Charles fine sandy loam, 1929. No fertilizer, 1926, 1927; 100 pounds of superphosphate in 1928; 200 pounds of 18 per cent superphosphate in 1929. Troup, No. 2—Susquehanna fine sandy loam. No fertilizer in 1926; 240 pounds superphosphate, 100 pounds of cottonseed meal and 6O pounds of nitrate of soda in 1927 ; 300 pounds of superphosphate, 100 pounds of nitrate of soda at planting; 150 pounds of nitrate of soda as side dressing in 1928; 200 pounds of 16 per cent super- phosphate and 100 pounds of nitrate of soda in 1929. Temple, No. 5—Houston clay, no fertilizer. Weslaco, No. 15—Victoria fine sandy loam, no fertilizer. College Station (Main Station)—Lufkin fine sandy loam; 300 pounds of 3-12-3 in 1926, 1927, 1928; 300 pounds of 3-10-3 in 1929. Lubbock, N0. 8—Amaril1o fine sandy loam, 1926. Richfield fine sandy loam, 1927 and 1929; no fertilizer. 12 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Denton, No. 6—San Saba clay; no fertilizer. a Chillicothe, No. 12—Kirkland clay loam, 1928. Vernon fine sandy. 10am, 1929. N0 fertilizer. A Iowa Park, No. 16——Yahola very fine sandy loam, 1926. Miller loam, 1927. No fertilizer. . Beeville, No. 1—Goliad fine sandy loam, 1926, 1927, 1929. Poesta fine sandy clay loam, 1928. RESULTS WITH THE RATION METHOD Table 2 contains the detailed results of some of the experiments-i while average results are given in Table 3. The results are summarized in Table 4., which gives the average gain in weight of the rats for the period of 8 to 11 Weeks, if they survived that long. These results show ' that white corn contains less vitamin A than yellow corn, and that straw; berry corn is intermediate between the two. It also shows that white corn grown at some places contains more vitamin A than that grown in‘ other localities. In some cases the white corn seemed to give as good a; growth as the yellow, but this may be due to defects of the method,» especially as the young rats. are known to contain vitamin A. The Chisholm white corn from Beaumont gave growth exceeding that of all except one of the samples of Ferguson yellow dent. Surcropper (white)? from Beaumont gave slightly more growth than Ferguson yellow dent from Beeville. Fentress strawberry (variegated) from Beaumont, Den-j ton, and Troup gave ‘more growth than most of the samples of Ferguson yellow dent. The Beaumont and Denton samples in general were more potent than the others. 1 These results will be mentioned further in connection with the dis- cussion of the results secured in subsequent years, by the use» of the," unit method. The more accurate results obtained by unit method did not confirm those results obtained by the ration method, which had a, assigned a high vitamin A content to white corn. i THE UNIT METHOD The ration method has the advantage of following the usual procedure? of feeding animals, but the amount eaten of the feed to be tested varies; with the appetite of the animal and also with its capacity to eat. As the animal grows larger it eats a larger quantity of food each day and thus; receives a larger amount of vitamin A. The quantity of vitamin A fed: varies with the appetite of the animal. Likewise, rats contain a consid» erable store of vitamin A at the beginning of the test and this aids in~ the growth of the rat. The amount of vitamin A stored in the varies with the character of the feed fed, and with the individuality of; the animal. ' The method developed by Sherman and Munsell (here termed the, unit method) (13, see also 8 and 15) was studied for the purpose of’ applying it to this particular problem. This method consists in feeding; 13 VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN .UOi-.w®s QOSNh 05H GMOU Hi < Chflhflah? GO 3a“ ~N5Uh>hmv-h mwGUEhhQQKQ 03w m0 050m m0 Qhflflk ... ....s.§!.w. . ... .-........ - -. x.¢ .- . . . - . . .. . ... ...... . . . . . . . ... . . . . . . . . . . .........Q%fl-HU>< I2 Gama QMOmVUNQQ I m4» om mm Ne. . .. . . . . . . . . . aaxah. QhSOhH. dawcoI/N/ wit’? QEOJMThC 2 . . . . . - . .@.NUQ @. @ . . . . . . . . . . . . .MNNQP .Q§Q.MH -.mmw%vflc>s Naavgflhxo h Gama ouomvwaofl oI m . m mm an am . . . . . . . . . . . . aaxah. dash. Qawqok/ wbbk/ abbobahbc m Aw»? Qéeaaao a a . m. 3 8 3 . . . . . . . . . . . . .228. défi. .552,» BE? “Eeazao wfima . . . . . . . .. IYNI wI . . . . . . . . ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 'lowmihw>< I2 . . . . . IaEaoZ NI I42 BI com ma . . . . . . . . . . . . . . . . Qaxwh. dsobh. Jmhhwflk/Nhww wmabcah 2 . . hmEhOZ NI wJl P: mIN wm . . . . . . . . . . . . . . . . aaxoh. dnobh. SbbwnBabw wmohbcoh h . . haEboZ NI #2 maa m: am . . . . . . . . . . . . . . . . iaxoh. n.Q:ohIH\ Qasaafiaw wwobaah h haEboZ NI I. I a2 a2 S“ . . . . . . . . . . . . . . . . aawah. dsohfi sksaaabw mwabcah hmhmm .. . . . . . . . . . . . . . .. wd woI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........ama._a>< h bah I ow wm wmI ow . . . . . . . . . . . . . . . . . . . nlwwxaFhagolHrfififHozoasm hwoobm h bah I 9m woI m: mm . . . . . . . . . . . . . . . . . . . IwaxoF dsobh. uwbfismh 352m h . .. bah I wd noI w: mm . . . . . . . . . . . . . . . . . . . Imaxoh. Ahsofifi Qwbfinh 3305 h . . . . ..... bah I o. I II owI mm . . . . . . . . . . . . . . . . . . . l¢mflxwk dsobw Qwbfismh 3.02m wmhmm . . . . . . . .. I6 In .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ......oma~a>< IQ . . . . . . ..@NQO © .. . . . . . . . . . . . . . . . . . . . . . . . .mNKQr%\ .Q5QHP .hOQQQhOhn.-@ m - WQ~%W(L AL» . x . . . . . . . . . . . . . . . . . . . . . . . . . wmwvmmvr? Xhiohrél ~hmwQQQRUh=% m . . . . . ..~. .tNUQ . . . . . . . . . . . . . . . . . . . . . . . . .MGNQP ghmwohk IHQQQQHOHfi-Z rv% wvflnvc w . . . . . . . . . . . . . . . . . . . . . . . . . .mflun®r_w JMUOhP .h®QQQhOh5@ .. . . . . . . . . . . .. .. . v.2 II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..owabo>< h . . . . . . . . . Qooh I a. NI NII NwI ow . . . . . . . . . . . . . . daxah. dnobh. JcoQ 302w? comswboh m . . . . . . . . . fmocm . . . . . . . . . . . . . . .wuwvwQP _Q5Q.~P “HQUQ P/Ohhnvw cQmfi-whvwvé h . . . . . . . . Ibooh I ob Nw <2 Na . . . . . . . . . . . . . Iwaxvh. dsobh. JcoQ Boza? comsmboh h . . . . . . . - .tflmva . . . . . . . . . . . . . . .WNNUF nQiohF Jwgnvg BOZU? gow:@hmwm h . . . . . . IaEIoZ I w. I mmI wwI mm . . . . . . . . . . . . . . daxoh. dzobh. JnaG 302w? comswboh h . . . . . . haEboZ I f: a: a2 S. . . . . . . . . . . . . . . éaxah. dsobh. 56G ao__sfi.m=a.sh Ehma . . . . . . . . . . . . . . .. mia a2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ............am_a.6>< m . . . . . . . édekoz @.% . . . . . . . . . . . . . . . . . .. . . WNKQF “Qsohm? Rvihhohm mwqhwwfl%% m . . . . . . ._NEHOZ . . . . . . . . . . . . . . . . . . . .wNNULw .Q§Qhr@~ Rvhrhzfivhg mwflhwwflw% h . . . . . - . hNEHoZ R..% . . . . . . . . . . . . . . . . . . . . WNNQF “QH-OMP .U_,.:@Qhm mwghwwfl: h . . . . . . . TWEHQZ @. . . . . . . .. . . . . . . . . . . . . . WNXQP QQQQMF anihhhohm W@QhaWNT% wxoa? mEflhw wEabm wEabm mEabm dZ 5w biaaI datum bu»? 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' . . . v . . . . . . . . . . . .. @.m ...........AQQ.@£3Q Ua@wohm w%gw#wfim I dZ 3 dZ Acoflfiw . m dZ 2,52% vial w .02 w .oZ QEEV w dZ v .oZ m. .oZ we Eou Mo fioifi/ 55C. $854 n33 xuosnsq c0252 awxfifim "Er/wow acofizmwm c32mc< o ozoU 6N2 E wcofifiwnsm mnoia.» i“ c33m Eco 5 2. aicmxi xwwB “on wcbfiw E 83 mo swBohm wmwi>\»,~ .. . o. s; -‘."‘>w‘_1.¢:.:\d1 a ration free from vitamin A but complete in other respects, until supply of vitamin A stored in the animal is exhausted, and then feedin weighed quantities of the feed to be tested in connection with the ration free from vitamin A. The amount of supplemental food Which causes an average growtl of 3 grams a week for 8 weeks, is said to contain a unit of vitamin This unit may be described as an average increase in weight of 3 gray, a week over a period of eight Weeks, following depletion of the bodily store of vitamin A. The problem is to determine the amount of corn or other feed Which, when fed daily for six days a week, enables the rat to, maintain one unit of growth or 24 grams in 8 weeks. That amount “i corn contains one unit of vitamin A. It was necessary to feed a basal ration so that the rat was certain to receive sufficient protein, energy, and enough of the essential minerals and vitamins other than vitamin A, in order to make sure that the gain in weight might be attributed only to the vitamin A present in l the daily portion of corn. The Sherman diet deficient in vitamin A (Mixture 666,), was first‘ used. This mixture is as follows: . Starch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .130 grams 3' Casein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 40 grams Yeast . . . . . . . . . . . . , . . . . . . . . . . . . . . . . . . . . . . . . . . 20 grams Salt mixture 7015 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 grams _,_ Sodium chloride . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2 grams The extraction of washed casein with a mixture of ether and alcohol 7 to remove vitamin A was both costly and laborious. Destruction of vitamin A by oxidation appeared to offer a simpler method and one which has proved successful (6a). To purify the casein, it was spread on trays in an electric oven to a depth of one-fourth inch and heated to 110 degrees C. for 24 hours. Air was drawn over the material by means of an electric fan and it was stirred from time to time during the day. The casein inactivated by oxidation was compared with casein purified j; by extraction, by means of rats fed the mixture given above. Results is were satisfactory. The commercial starch also contained appreciable amounts of vitamin A. Untreated starch, starch extracted with a mixture of ether and. alcohol, and starch purified by oxidation were compared by means of rats fed on the mixture mentioned above. Destruction of vitamin A by - a oxidation was found to be satisfactory. ; Steenbock and {others have emphasized the necessity of supplying T vitamin D in the ration. Experiments were made using corn with irradiated Wesson oil up to the time of cessation of growth and the appearance of symptoms of deficiency in vitamin A. The irradiation had no efiect on growth, but since rachitic symptoms may develop at a later date, and to avoid any possibility of deficiency in vitamin D, y irradiation of the basal ration was used. \.;-1'¢§~,q.-,¢l,,._<., t, s...».,,sl_{g VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 19 The method used was as follows: Two to six rats 23 to 26 days old were placed in a round wire cage (tinned test-tube basket) with wire screen bottoms and fed Mixture 666. In 4 to 5 weeks the rats usually showed symptoms of deficiency. The first indication was generally a slight accumulation of exudate in the cor11er of one or both eyes. This was followed by swollen and reddened eyelids and later by extensive exudate. With the appearance of definite xerophthalmia, maintenance or loss in weight was observed. At this time, usually about the thirty- fifth day, the rats were placed in individual cages. When the rats stopped gaining weight, or otherwise showed indications of deficiency of vitamin A, weighed quantities of the feed to be tested were fed daily for eight weeks, unless the rat died before the end of the period. Considerable experience and care are necessary to know when the rats are ready to be put on test of the feed to be assayed for vitamin A. If the rats are taken too soon, the results will be unsatisfactory. On the other hand, if they are taken too late, advanced pathologic conditions may be established. Generalized infections resulting in the formation of pus in the upper part of the digestive and respiratory tract appar- “ently cannot be cured by feeding methods, regardless of the richness of the diet in vitamin A. Lung trouble is a weakness to which the rat is very susceptible; in fact, a big majority die of it; consequently respira- tory infection may be the result of radical organic disability and not due to deficiency at all. Although the Albino rats used were supposedly homozygous, there was considerable variation in growth. This variation has been recog- nized by the majority of investigators and is variously attributed to the capacity of the individual to grow, to pathological lesions not attributable to the diet, to the varying amount of feed eaten, to seasonal variation, and perhaps other causes. Attempts were made to feed such a quantity of material containing vitamin A that the rat made an average gain of 3 grams per week for 8 weeks. It is difficult to estimate the amount of feed required to pro- duce exactly this gain. Usually the estimate of the amount of vitamin A present in the food tested was made from gains near that desired; that is, from one lot gaining less than this amount, and another more than the 3 grams a week. When the rats gained less than 3 grams a week, many of them would die before the period was over. If the average of gains in weight was too far from that desired, it was neces- sary to repeat the experiment, feeding larger or smaller quantities of the feed to be tested. These repetitions were» frequently needed, on account of the difficulty of judging closely the exact amount of the food which should be fed to secure the gain of 3 grams a week. The use of six or more rats in each test was found desirable. Some of them usually died before the test was ended. A sudden drop in weight fol- lowed by death was probably not due to deficiency of vitamin A. It was desirable also to make check tests beginning on different days; the results secured at different times did not always agree. 20 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION The growth of the rats, up to 6 grams a Week, was sometimes found t0 be in proportion to the quantity of vitamin A fed, but in other cases y the rats made unusually large gains When fed a little more than one unit of vitamin A. Thisis the chief reason Why it is desirable to esti- mate the quantity of vitamin A in the food from the results from two groups of rats, one gaining less than 3 grams a Week during the eight Weeks, and the other a little more than 3 grams. It is also necessary to consider Whether the rats which died suffered from deficiency in . vitamin A or from other troubles. In spite of all precautions, however, the results are probably accurate only to 1‘0 to 20 per cent. Comparison of the Unit Method and the Ration Method Some of the samples of corn were tested by the ration method and also by the unit method. The results are compared in Table 5. The corn is tabulated in order of content of vitamin A by the unit method, beginning with those samples containing the highest amount. On an average the order is similar, but with some individual samples, the order is decidedly different. The sample which comes first by the unit method, comes third by the ration method. The sample which comes second by the unit method is decidedly the highest by the ration method. Table 5.—Comparison of unit method for vitamin A with the ration method. Average Grams corn Units weekly Variety t0_ one unit vitamin A gain of rats _, vitamin A, to one gram 1n grams, is unit method corn. ration i unit method method 1 Ferguson Yellow Dent corn . . . . . . . . . . . . . . . . . . . . . 0.3 3.3 10.9 Ferguson Yellow Dent corn . . . . . . . . . . . . . . . . . . . . . 0.35 3 .0 14.6 Ferguson Yellow Dent corn . . . . . . . . . . . . . . . . . . . . . 0.4 2.5 11.0 Fentress Strawberry . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0.6 1 .7 10.7 _ Fentress Strawberry . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0.8 1 .3 8.9 Fentress Strawberry . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 .1 .9 7.2 Brazos White . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 +‘? .25? 8.4 Mexican June (white) . . . . . . . . . . . . . . . . . . . . . . . . . . 15 + .07— 5.3 ~ Hastings Prolific (white) . . . . . . . . . . . . . . . . . . . . . . . . 25 + 04— 3.0 ‘t Oklahoma White Wonder . . . . . . . . . . . . . . . . . . . . . . . 25 + .O4— 6.1 ~ Horton (white) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 + .04—— 7.3 " Hastin s Prolific (white) . . . . . . . . . . . . . . . . . . . . . . . . 35 + .03— 5.6 ' , Chisho m (white) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 +? 0 4.4 , Blue Grain (white) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 +? 0 5.3 The Sherman-Munsell unit method is decidedly superior to the ration method, since it brings out more clearly the difierences in the vitamin A content of the various samples, and provides a much more accurate method for measuring the vitamin A in foods. It requires more time, care, and rats, but the superior accuracy justifies the additional Work. Modification of the Ration for Corn Low in Vitamin A The samples of White corn were so low in vitamin A thatthe rats could not eat daily a quantity containing one unit. In order to secure VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 21 ingestion of a unit of vitamin per day, the White corn to be tested was added to a mixture containing vitamin A in the form of yellow corn. This mixture, No. 685, consisted of the following: Per cent Casein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Salt mixture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . '. . . . . 4 Yeast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5 Irradiated Wesson oil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Yellow corn (29229) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 3.75 Added test feed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .71.25 When 4 grams of this mixture were fed each day, the rat received .15 gram of yellow corn, which amount was found to contain about 1.0 unit of vitamin A. In addition, the rat received 2.85 grams of the food to be tested in each 4 grams of the mixture. Additional gain above that due to the yellow corn was attributed to the vitamin in the added feed being tested. The estimate of the quantity of vitamin A was made from the results of the test. On account of the small quantity of vitamin A present and the difficulty of measuring it, the number of tests on vitamin A in white corn was restricted, and the results are only approximate. RESULTS OF TESTS WITH THE UNIT METHOD Detailed results of some of the experiments are given_in Table 6. Records were also made of the condition of the animals and the amount of feed eaten. Average results for a number of the experiments are given in Table 7. Vitamin A in Yellow Corn The estimated quantity of corn which contains one unit of vitamin A is summarized for Ferguson Yellow Dent corn in Table 8. The amount of corn containing one unit of vitamin A varied from 0.14 to 0.4 gram; or, to put it another way, one gram of the yellow corn contained 2.5 to 7 units of vitamin A. The corn grown in 1926 contained less vitamin A than in 1927, and this, in turn, less than in 1928, though the differences between 1927 and 1928 were usually small. With one exception (Angle- ton) there was little difference in the vitamin A content of corn grown in the various places in 1928. In. 1927, corn grown at Angleton, Nacogdoches and Temple contained less vitamin A than the others. In 1928, Angleton corn still had the lowest content of vitamin A. Both the season and the locality appeared to have some effect on the vitamin A content of the yellow corn. The vitamin A in varieties of yellow corn other than Ferguson Yellow Dent was also determined. The results are given in Table 9. The amount of corn which contains one unit of vitamin A varies from .13 to 0.2 gram; or one gram of corn contains 5 to '8 units. This variation is comparatively small. The other varieties of corn tested contained about the same amount of vitamin A as does Ferguson Yellow Dent. fiiiuwilqfiywa, v 22 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION .3! ew w“ N“ ““ ww ww ew ww ww “ . wl. ew N“ ““ w“ e“ w“ w“ e“ w“ “. 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Variety and location 1 of rats at be- gain in life,- daily begun ginning weight weeks 25859 Ferguson Yellow Dent, Angleton, Tex. .5 2 71 5 3 1.0 4 86 71 _ 7.5 1 .5 2 77 129 8 2 3 92 52 8 25840 Ferguson Yellow Dent, BeaumonLTex. .3 6 87 4 5 .3 .6 2 65 88 8 1 .0 2 65 83 8 1 .2 2 64 68 8 ~ 1 .5 2 71 113 8 25932 Ferguson Yellow Dent, Beeville, Tex. .2 2 67 7 5 . 3 4 86 24 6. 8 .6 3 75 59 8 1 .O 5 63 66 7 . 6 2 2 56 84 8 1W ix 106 ~ 4 3 68 103 8 26142 Ferguson Yellow Dent, Lubbock, Tex. .3 4 95 . . . . . . . . ' 5.8 .35 3 69 —9 4 .4 6 95 28 7 1 .0 5 64 76 7 1 .5 2 69 74 8 4 3 76 5O 7 28636 Ferguson Yellow Dent, Angle-ton, Tex. .2 ' 6 109 16 5.7 ~ .3 3 69 15 8 .35 4 74 43 4 . 4 2 74 62 8 .6 2 52 71 8 .8 2 52 9O 8 1 2 67 67 8‘ _ . 1 .5 2 74 107 8 28645 Ferguson Yellow Dent, Troup, Texas. .2 9 87 37 8 .4 2 65 143 8 .6 2 52 7O 8 .8 2 68 78 7 1 .0 2 62 109 8 1 . 5 2 71 126 8 28651 Ferguson Yellow Dent, Denton, Tex. .2 7 88 25 6.9 .25 4 91 . . . . . . . . ’5.8 .3 2 74 O 6 .35 4 66 55 8 . 4 2 83 114 8 .8 2 76 73 7 1 2 73 6O 8 1 .5 2 55 71 8 28983 Ferguson Yellow_Dent, Nacogdoches, .2 8 93 —-9 6.3 Te-Xas- .3 4 53 15 7.5 .35 3 79 28 8 .4 2 73 9O 8 . 8 3 7O 89 7 . 7 1 2 69 76 8 1.2 2 83 65 8 29032 Ferguson Yellow Dent, Lubbock, Tex. .2 2 97 18 5.5 .3 6 91 ‘ 3O 8 x35 4 56 88 32 .4 6 9O , 47 7.8 .8 2 74 100 7 _ 1 2 75 ~ 74 8 29038 Ferguson Yellow Dent, Beeville, Tex. .2 7 87 29 7.5 .3 4 78 52 8 .4 2 67 76 8 1 2 67 67 8 1 .2 2 69 94 8 , w‘ I VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 27 i Table 7.—Average results from feeding tests by the unit method for vitamin A——Continued. Average _ Grams Number weight Average Average No. Variety and location fed of rats at b_e- gain in life, daily begun ginning weight weeks 29050 Oklahoma Yellow Dent, lowa Park. .1 4 81 2 8 . 15 5 75 5 6 .4 .2 3 74 41 8 .4 2 66 81 8 .6 2 63 59 8 _ 1 2 71 101 8 29142 Ferguson Yellow Dent, College Station. .2 5 87 28 8 ._5 2 65 81 7 29229 Yellow Corn, College Station . . . . . . . . .15 3 72 28 3 .2 15 84 36 6.5 .3 7 84 47 8 . 4 3 77 93 8 . 6 4 73 75 8 .8 10 76 97 8 y 1 .0 4 83 81 8 1 .2 4 83 101 8 30214 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 6 83 29 8 .2 3 57 48 7 . 3 .3 3 53 62 8 30215 Golden Harvest, Troup . . . . . . . . . . . . . .1 3 78 18 7.3 . 15 4 69 63 6.3 .2 4 57 55 7 . 5 .3 4 60 60 8 30216 Revters Improved Golden, Troup. . . . .12 3 70 5 8 .15 4 77 47 6.5 .2 2 49 60 8 .3 4 66 64 8 30217 Ferguson Yellow Dent, Troup . . . . . . . . .13 6 72 9 8 .15 4 77 41 7.3 .2 2 61 42 8 .3 4 56 60 i; 30218 Ferguson Yellow Dent, pollinated by .15 7 83 36 6.6 Ferguson Yellow Dent, deep yellow .18 4 77 45 7.5 color, three factors for endosperm .2 4 79 3 6 color. .3 3 64 53 8 30219 Ferguson Yellow Dent, Pollinated by .15 4 64 21 7 Surcropper, dilute yellow, two factors .2 4 80 4 6 .5 for endosperm color. .25 4 64 62 7 .5 .3 2 68 42 8 .4 2 66 50 8 30220 Surcropper, pollinated by Ferguson 2 .0 2 64 89 ' 8 Yellow Dent, one factor for endo- .3 4 70 21 7 sperm color. .4 4 83 .8 5.3 ‘ .5 2 70 66 8 1 2 64 78 8 30222 Yellow Dent X SurcroppenF. hybrid—~ . 12 4 56 11 3 deep yellow, three factors for endo- .14 4 67 17 7 .5 sperm color. . 18 4 62 62 8 .2 4 73 21 8 .3 4 69 54 8 30223 Yellow Dent X Surcropper, F. hybrid, . 15 4 61 7 6 dilute and pale yellow, one and two .2 9 75 94 6 .2 factors for endosperm color. .25 4 63 30 6.3 .3 4 63 46 _ 6 .4 2 73 54 i 7 30270 Nicholson's Giant Yellow, Beeville, .2 5 79 31 6.2 Texas. .3 4 60 39 l 7 28 Table 7.——Average results from feeding tests by the unit method for vitamin A—Continued. BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION ,. 61 Average '»_ Grams Number weight Average Average i‘ No. Variety and location of rats at be- gain in life, ’ daily begun ginning weight weeks ~ 30271 Wo0d’s Golden Harvest, Beeville, .12 4 69 17 8 .1 Texas. .15 8 79 47 5.7 '_ .2 4 89 34 6.3 - .3 4 63 63 8 30272 Reutefs Improved Golden Dent, Bee- .12 4 65 15 7.3 .. ville, Texas. .15 4 75 45 6.3 _ .2 2 71 57 8 '. .3 4 67 7O 8 , 30273 Ferguson Yellow Dent, Beeville, .15 6 77 2s 5.5 7. Texas. .2 10 78 33 6.5 , .3 4 74 51 7.3 _, 30310 Yellow Dent, Temple, Texas . . . . . . .. .14 4 68 21 5.5 é . . .2 5 57 48 7.2 A_ 30373 Pilgrim Yellow Dent, Denton, Texas. .15 4 69 36 7 .2 4 75 33 8 30385 Bloody Butcher, Denton, Texas . . . . . . _15 4 63 5 8 .2 3 75 40 8 .7 2 93 83 8 . .8 2 66 106 5.5 .1 30386 Wright Yellow Dent, Denton, Texas. . _12 3 83 14 6 v .15 3 85 ' 28 7.3 .2 2 58 5O 2 - 30481 Ferguson Yellow Dent . . . . . . . . . . . . . . .2 5 78 28 6. 30482 Yellow Creole, Nacogdoches, Texas. . . _12 4 89 3 2.8 .15 3 75 41 7.3 _ .2 4 68 50 7 30483 Yellow Creole, Angleton, Texas . . . . . . _ 12 4 87 15 5.5 .15 6 73 30 8 .2 '2 54 53 8 30484 Ferguson Yellow Dent, Angleton, Tex. .2 4 62 2 6.3 .22 2 74 27 5 .25 4 66 27 7.5 .4 4 83 -—-10 6.5 30491 Yellow Creole, Main Station Farm. . . .15 6 74 36 7 < .2 6 79 24 7.7 30492 Ferguson Yellow Dent, Main Station .15 6 73 25 6.5 Farm. 2 2 62 57 8 30493 Ferguson Yellow Dent, Weslaco, Texas . 14 4 78 29 7 2 2 73 51 8 30494 Yellow Creole, Weslaco, Texas . . . . . . . .15 4 62 29 7.5 .2 4 82 37 8 30496 Improved Golden, Weslaco, Texas. . . . . 15 6 82 10 5.2 . ' .2 4 81 49 5.8 30720 Oklahoma Yellow Dent, Iowa‘ Park, .15 4 70 33 8 Texas. .2 5 69 43 30721 Ferguson Yellow Dent, Iowa Park, .15 3 75 29 7 Texas. .2 3 63 50 7.3 30224 White (Yellow Dent X Surcropper) In 685 Division of Agronomy, F. hybrid, 2.5 4 6O 3 6 no factors for endosperm color. In 685 3 4 67 23 In 685 ' 4 60 6.5 VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 29 Table 7 .—Average results from feeding tests by the unit method for vitamin A—Continued. Average _ _ Grams Number weight Average Average N0. Variety and location of rats at be- gain 111 lif daily begun ginning weight weeks 30997 Yellow Corn, Illinois . . . . . . . . . . . . . .. .1 4 81 16 7 .12 4 69 12 6.5 .15 6 80 ——5 6.9 .18 5 79 17 7.2 .2 4 67 49 8 31944 Ferguson Yellow Dent, pollinated by .12 6 61 25 7.4 Ferguson Yellow Dent, deep yellow, . 15 8 72 42 8 three factors for endosperm color. .2 4 91 55 8 31950 F. hybrid, Ferguson Yellow Dent X .3 4 77 6 5. Sureropper, pale yellow, one factor .4 4 93 2 6 for endosperm color. .5 4 80 . . . . , . . . 5.7 .7 4 74 15 3-.5 .9 5 69 35 8 31951 F‘. hybrid, Ferguson Yellow Dent X Mix 685 Surcropper-White, no factors for .4 4 56 . . . . . . . . 5 endosperm color. Mix 687 .4 4 76 . . . . . . . . 4.3 30994 Yellow Corn Germ, Illinois . . . . . . . . . . 1 2 71 8 4 ' .15 6 71 14 6.2 .2 8 75 . . . . . . . . 5.3 .3 4 77 . . . . . . . . 5 .4 4 79 15 3. .5 5 88 12 5 1 .0 2 83 . . . . . . . . 1 31945 Ferguson Yellow Dent, pollinated by .16 2 95 17 7.5 Surcropper, dilute yellow, two fac- .2 7 74 29 7.8 tors for endosperm color. 31946 Surcroper, pollinated by Ferguson 685 Yellow Dent, pale yellow, one fac- +.3 4 76 8 - 6.3 tor for endosperm color. .4 8 77 18 6. 5 .45 4 92 13 5.5 55 5 5O 40 7.8 7 4 107 11 8 31947 Surcropper, white, pollinated by Sur- 4 1 48 22 8 cropper, no factors for endosperm Mix 685 color. 4 1 75 . . . . . . . . 7 Mix 687 4 4 80 . . . . . . . . 4. 8 31948 F. hybrid, Ferguson Yellow Dent X .3 4 94 15 8 Surcropper, deep yellow, three fac- .14 4 71 . . . . . . . . 6 tors for endosperm color. .2 6 87 24 7. 7 .25 7 89 35 7.6 31949 F. hybrid, Ferguson ‘Yellow Dent X .3 10 89 18 6.8 Surcropper, dilute yellow, two fac- .4 9 73 33 7.1 tors for endosperm color. Vitamin A in Variegated and in Red Corn One variety of corn, Fentress Strawberry, with variegated or “calico” pericarp, and one with red pericarp, Bloody Butcher, were tested. -The results for 18 samples are summarized in Table 10. The amount of variegated corn which contained one unit of vitamin A. varied from .35 to 1.1 grams; or one gram of corn contained 2.9 to 0.9 units of vitamin A. The red corn contained 1.8 to 5.5 units of vitamin A per gram. 30 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Table 8.—Approximate vitamin A in Ferguson Yellow Dent. Units of vitamin A Grams corn to one unit in one gram corn Where grown - 1926 1927 1928 1926 1927 1928 Angleton . . . . . . . . . . . . . . . . . . . . . . .. 0.4 .35 .25 2.5 2.9 4.0 Beaumont . . . . . . . . . . . . . . . . . . . . . . . .35 . . . . . . . . . . . . . . . . 3.0 . . . . . . . . . . . . . . .. Beeville . . . . . . . . . . . . . . . . . . . . . . . .. .3 .20 .18 ,/ 3.3 5.0 5.5 Lubbock . . . . . . . . . . . . . . . . . . . . . . .. .4 .24 . . . . . . .. 2.5 4.0 . . . . . . .. Troup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .16 .14 . . . . . . .. 6.0 7.1 Denton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .20 . . . . . . . . . . . . . . .. 5.0 . . . . . . .. Nacogdoches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35 . . . . . . . . . . . . . . . . 2.9 . . . . . . . . College Station . . . . . . . . . . . . . . . . . . .28 .18 .15 3.6 5.5 6.7 Temple . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .28 .15 . . . . . . .. 3.6 6.7 Weslaco . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .15 . . . . . . . . . . . . . . .. 6.7 Iowa Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .14 . . . . . . . . . . . . . . .. 7.1 Table 9.——Vitamin A in varieties of yellow corn. Units _ Grams corn vitamin A No. ' Variety and where grown to one to one gram unit corn 29050 Oklahoma Yellow Dent, Substation No. 16, Iowa Park. . . .18 5.5 29229 Yellow corn. College Station . . . . . . . . . . . . . . . . . . . . . . . . . . .15 6.6 30214 Giant Yellow, Substation No. 2, Troup . . . . . . . . . . . . . . . . . .15 6.6 30215 Golden Harvest, Substation No. 2, Troup . . . . . . . . . . . . . . .13 7.7 30216 Reuter’s Improved Golden, Substation No. 2, Troup.. . . . .14 7.2 30270 Nicholson’s Giant Yellow, Substation No. l, Beeville. . . . . 18 5.5 30271 Wood’s Golden Harvest, Substation N0. 1, Beeville . . . . . . .14 7.2 30272 Reuterils Improved Golden Dent, Substation No. 1, Bee- 14 7 2 vi e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 30373 Pilgrim Yellow Dent, Substation No. 6, Denton._ . . . . . . . . . 14 7.2 30385 Bloody ButEher, Substation No. 6, Denton . . . . . . . . . . . . .18 5.5 0386 Wright Yellow Dent, Substation No. 6, Denton. . . . . . . .15 6.6 ' 30481 Ferguson Yeoolw Dent, Nacogdoches . . . . . . . . . . . . . . . . . , 5.0 30482 Yellow Creole, Substation No. 11, Nacogdoches. . . . . . . .14 7.2 ..» 30483 Yellow Creole, Angleton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 6.6 § 30491 Yellow Creole corn, College Station . . . . . . . . . . . . . . . . . . . . .15 6.6 < 30494 Yellow Creole, Substation No. 15, Weslaco . . . . . . . . . . . . . .15 6.6 Q. 30496 Improved Golden, Substation No. 15, Weslaco . . . . . . . . . . .2 5.0 . 0'12 Oklahoma Yellow Dent, Iowa Park. . . ._ . . . . . . ._. . . . . . . . . .14 7.2 i 30997 Yellow Corn, The Quaker Oats Co., Chicago. Illinois. . . . . .2 5.0 Table 10.—Vitamin A in Strawberry corn and Bloody Butcher corn. Grams corn to one unit vitamin A Units vitamin A to one gram corn Where grown 1926 1927 1928 1928 1926 1927 1928 1928 in Texas Straw- Straw- Straw- Bloody Straw- Straw- Straw- Bloody berry berry berry Butcher berry berry berry Butcher IowaPark . . . . .. .6 . . . . . . . . . . . . . . .. .55 .7 . . . . . . . . . . . . . . .. 1.8 Beeville . . . . . . .. .8 .7 . . . . . . .. .35 1.2 1.4 . . . . . . .. 2.9 College Station. 1.1 .7 .5 . . . . . . .. 0.9 1.4 .0 . . . . . . .. Angleton . . . . . . . . . . . . . .. .9 .7 . . . . . . . . . . . . . . .. 1.1 1.4 . . . . . . .. Troup . . . . . . . . . . . . . . . . .. .9 .4 . . . . . . . . . . . . . . .. 1.1 .5 . . . . . Nacogdoches . . . . . . . . . . . . .85 . . . . . . . . . . . . . . . . . . . . . . . . 1.2 . . . . . . . . . . . . . . . . Lubbock . . . . . . . . . . . . . .. 1.0 . . . . . . . . . . . . . . . . . . . . . . .. 1.0 . . . . . . . . . . . . . . .. Denton . . . . . . . . . . . . . . . . . . . . . . .. .35 .18 . . . . . . . . . . . . . . .. 2.9 5.5 Weslaco . . . . . . . . . . . . . . . . . . . . . . .. 1.0 . . . . . . . . . . . . . . . . . . . . . . .. 1.0 . . . . . . .. Temple . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .18 . . . . . . . . . . . . . . . . . . . . . . .. 5.5 VARIATIONS IN’ VITAMIN A AND CHEMICAL COMPOSITION OF CORN 31 Both the variegated corn and the red corn Were more variable than any variety of the yellow corn. The reason for the lower vitamin con- tent and greater variability in vitamin content of red and of variegated corn as compared to yellow corn probably lies in the fact that neither of the varieties used is homozygous for endosperm color. The red color of the pericarp obscures the color of the endosperm so that varieties with colored pericarp. may become mixed in their endosperm color. The Division of Agronomy determined the endosperm color on 56 ‘self- pollinated ears of Fentress Strawberry, the variegated variety, and found that 5-3.1 per cent of the plants were white-seeded, 32.1 per cent yellow- seeded, and 12.5 per cent were segregating. If vitamin content is com- pletelyassociated with yellow color this variety should have about 38 per cent as much vitamin as it would have if it were pure yellow. a Samples of this variety which are known to be pure for yellow endosperm are now being tested in comparison with samples known to be pure for white endosperm. In the meantime it may be of interest to note that the vitamin content of Strawberry corn in comparison with Ferguson Yellow Dent grown at the same stations was 30.5 per cent, 27.4 per cent, and 28.2 per cent, respectively, for the years, 1927, 1928, and 1929. As with the yellow corn, the red corn grown in 1928 contained the highest amounts of vitamin A, that in 1927 next, and that grown in 1926 the lowest. ~ Vitamin A in White Corn As previously stated, the white corn was fed in a complete ration mixed with yellow corn to supply part of the vitamin A needed. Ap- proximate results, on 28 samples, are given in Table 11. The amount of vitamin A in most of the samples of white corn was very low, one unit in 25 to 35 grams. Some few samples, however, contained one unit in 2 to 5 grams of corn, which is high for white corn. This variation may possibly be due to crossing with the yellow corn. RELATION TO HEREDITY The yellow pigmentation in the endosperm of corn is known to be inherited in definite Mendelian ratios, and it is possible to produce samples of corn having O, 1, 2, or 3 genetic factors for yellow endosperm. Samples of corn of known genetic constitution with regard to endosperm color were furnished by Dr. P. C. Mangelsdorf of the Division of Agronomy (Gb) . The results are given in Table 12. In one test the four types were produced by pollinating Surcropper and Ferguson Yellow Dent by a mixture of pollen from both varieties. In the other test, four types were obtained from the same ears, which were on first- generation hybrids of Ferguson yellow dent and Surcropper, In the first test the units of vitamin A per gram of corn were approxi- mately in proportion to the number of genetic factors for yellow endosperm. Samples of corn involving 1, 2, and 3 genetic factorslcon- tained vitamin A in the proportions 2.5 :5 :7 in 1928 and 2 :5 :8 in 1929, the average for the two years being 2.25 :5.00 27.50. 32 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Table 11——Vitamin A in white corn _ Grams corn Units N0, Variety and where grown to_ one unit vitamin A vitamin A to one gram ‘ ' corn 25852 Hastings Prolific, Substation N0. 3, Angleton, Texas. . . . . 35 + .03 25853 Brazos White, SubstationNo. 3, Angleton, Texas... . . . . . 4‘? .25 28637 Hastings Prolific, Substation No. 3_, Angleton. Texas. . . . . 12 .08 28638 Oklahoma White Wonder, Substation No. 3, Angleton, , Texas...._ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 0 .00 25929 Oklahoma White Wonder, Beeville, Texas . . . . . . . . . . . . . . 25 + .04 25930 Hastings Prolific, Beeville, Texas. . ._ . . . . . . . . . . . . . . . . . . . 25 + .04 29039 Siircropper. Substation No. 1, Beeville, Texas . . . . . . . . . . . 5? .20 29040 Chisholm, Substation No._ 1, Beeville. Texas . . . . . . . . . . . . 22 .05 2904] -Hastings Prolific, Substation No. 1_, Beeville. . . . . _. . . . . . . 25 + .04 29042 Oklahoma White Wonder, Substation No. 1, Beeville, Texas . . . . . ._ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 + .03 25942 Horton. Substation No. 1_1, Nacogdoches, Texas . . . . . . . . . 25 + .04 28979 ' Hastings Prolific, Substation No. 11,_ Nacogdoches, Texas 3‘? .33 28980 Oklahoma White Wonder, Substation No. 11, Nacog- doches, Texas., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2? .50 28981 Siircropper, Substation No. ll, Nacogdoches, Texas. . . . . 25 + .04 28982 Chrisholm, Substation No. 11, Nacogdoches, Texas . . . . . . 2 .50 29031 Surcropper, Substation No. 8, Lubbock, Texas . . . . . . . . . . 28 .04 25923 Mexican June, Substation No. 16, Iowa Park, Texas. . . . . 15 .07 29051 lVIexican June, Substation No. 16, Iowa Park, Texas._. . . . 22 + .05 29135 Hastings Prolific, l\/1ain Station Farm, College Station, Texas . . . . . . . . . . . . . . . . . . . . . . . . . . 10 .10 29136 Nacogdoches, Main Station Farm, College Station, Texas. 3? .33 29137 Surcropper, Main Station Farm, College Station, Texas. . 2‘? .50 29138 Oklahoma White Wonder, Main Station Farm, College Station, Texas. . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . .. 22 + .05 29139 Brazos White, _Ma1n Station Farm, College Station, Texas. 17 .06 29140 Chisholm, Main Station Farm, College Station, Texas. . . .22 .05 29141 Thomas, Main Station Farm, College Station, Texas. . . . . 5 .50 29143 ITIOTLOIInR/IHXII Station Farm, College Station, Texas. . . . . 2 .50 29144 Va. White Dent, Main Station Farm, College Station, Tex. 22 + .04 Inthe second test in which all four types were produced on the ‘same ears the agreement in the ratio was not so close though quite evident. Also all types appeared to be lower in. vitamin A than the corresponding lot in the first test. VARIATIONS DUE TO SEASON AND LOCALITY It is evident from examination of Tables 8, 9, and 10, that the season has some effect upon the vitamin A content of corn. Both the yellow corn (Table 8) and the red corn, (Table 10) contained more vitamin A in 1928 than in 1927, and more in 1927 than in 1926. The differences were large in corn from some places and small at others. . Some influence of the locality is also" apparent. Yellow corn grown at Angleton contained (Table 8) less vitaminA than that grown in other, localities, in all three seasons. This difference is not so apparent with the red corn. The yellow corn and the red corn grown at Nacog- doches (one season) contained about the same amount as that- at Angle- ton. Corn grown at Lubbock was a little low in. vitamin A in the yellow variety one season, a, little less than the average in the yellow variety one season, and low in the red variety one season. Consistent relations between the amounts of vitamin A in the corn and the locality in which it was grown are difficult to trace. The differences observed may be partly due to seasonal conditions, and partly to cross pollination. 33 VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN n WHAQ. + @ . . . . . . . . . . . . . . . . . . . . . . HUQQQFOQQZ X HAMUQ 39.20% . . . . . . . . . . kUQQO.mOh:@ x PQQQ aoiQy m. ~ w. H . . . . . . . . . .25?» ‘Axum . . . . . . . Jnwnmoguksm X fiSC Eozfir . . . . . . . . . Qoaaohzsm X fiSD 30:“; omen m. mm. N . . . . . . . 450:9» ofiifi . . . . . . . Loaaohupsm X fiSQ 302w? . . . . . . . . . Qwnnopopsm X fiED Bozfiw mwmim v m. m . . . . . . . . .3211» 96G . . . . . . . Qoanohism X EQG Kozw? . . . . . . . . . Qwanowfism X fiED Bo:o> wvafim Amew. + © . . . . . . . . . . . . . . . . . . . . . . . . ... . . . _ . . . . . fmmvgacknwhsvx . . . . . . . . . . . . . . . . . . . . . . . .HUQQOQUMQ@ N m. H . . . . . . . 4 . .3029» 2mm . . . . . . . . . . . . . . . . . . . écwQ Eozw? . . . 4 . . . . . . . . . . . . . . . . . . . Qwaaohuhsm wvmfim fi §. x . . . . . . . .>?C@Tw% . . . . . . . . . . . . . . . . . . . . . . .HQQQOQU1:J@ . . . . . . . . . . . . . . . . . . . . _ . IWQUO gmvifly % . m . . . . . . . . . .>lo@émv~fi QUUQ . . . . . . . . . . . . . . . . . . . . QwQUQ Rrmvimy . . . . . . . . . . . . . . . . . . . . . . HQUQ R/Oiwuf 32 l||@o¢ + O . . . . . - . . . . . . . . . - . . . . . . . . . . . x . . . . . . 4 . . x w mm. m was _ 3oz?» 23 was waszfi . . . . . . . ..8@no.6.5m X 15D 3ozu> . . . . . . . . . Qvaaopupsw X flEO 3oz“; mmmom w f. m . . . . . . . . 55:2, QQQQ . . . . . . . Jmzmaohfism X 20G 30:8» . . . . . . . . . Lwamobhsw X EQQ 3o:@> mmmom @ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . - . . ..HQQQQ.~U.~:J4 . . . . . . . . . . . . . . . . . . . . . . . fhmvganvhnuhsi w . . . . . . . . . .~$O=Q> U-w@ . . . . . . . . . . . . . . . . . . . . flzwmwc RIOZU> . . . . . . . . . . . . . . . . . . . . . . . . kUQQOMUM§W AL. mw . . . . . . . .>/Q@@U»% . . . . . . . . . . . . . . . . . . . . . .¢~QQQOMOM:Z . . . . . . . . . . . . . . . . . . . . . ..HCQQ krozww N. .@ . . . . . . . . . 3Q@@U% QQUG . . . . . . . . . . . . . . . . . . . 2ro@%mu> . . . . . . . . . . . . . . . . . . . . . . .P§U@% 39:0? M5: EQQQmOCGQ 62w 4. c153? 30:9» .0 Z one E Mo .5“ $303 .630 Qcwpma c275 20.25 uvww ~33 4 55m»? in: 25 325w Afionmq BED .6“ wEwtU Mo .oZ .58 E .530 8.89635 3H E303 285% o» coin?! E < c323? we mtcDlfifi vEdF 34 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION CORN MEAL AND HOMIN Y FEED In the manufacture of corn meal for human use or for animal feed- ing, the corn is ground and part of the bran and germ removed. The , process varies, so that corn meal may range in character from a meal containing nearly all the grain, with little bran or germ removed, to a I‘ highly purified meal containing little bran or germ. The meal contain- ing little bran or germ is usually considered to be of higher commercial t quality than the other, and sells for a slightly higher price. Corn meal i for human use is usually made from white corn in the South, but in the 7 North it is frequently made from yellow corn. The use of yellow corn p. meal as a human food on account of its content of vitamin A would no doubt be desirable in many parts of the South, particularly for those it who live upon a restricted diet. Since cornmeal is made chiefly from the endosperm of corn, and the endosperm of yellow corn is rich in vitamin A, yellow corn meal should likewise be well supplied with vitamin A. _ Tests for vitamin A in corn meal are given in Table 13. Vitamin A in the first two lots (white corn) was determined by the ration method. There appear to» be larger amounts of vitamin A in the corn meal than 3e in the corn or corn bran or hominy feed. Table 13.—~Vitamin A in corn, corn meal, and milling products. Weekly Grams to Units gain of one unit vitamin A rats, vitamin A in grams White corn meal, unbolted (Palestine) . . . . . 7.2 . . . . . . . . . . . . . . . . . . . . . . . . Pearl meal from the above . . . . . . . . . . . . . . . . 12.8 . . . . . . . . . . . . . . . . . . . . . . . . Corn bran from the above . . . . . . . . . . . . . . . . 8.0 . . . . . . . . . . . . . . . . . . , . . . . . White corn (Dallas) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. Pearl meal from the above . . . . . . . . . . . . . . . . 7.9 . . . . . . . . . . . . . . . . . . . . . . .. La France meal from the above . . . . . . . . . . . 9.8 . . . . . . . . . . . . . . . . . . . . . . . _. Hominy feed from the above. . . . . . . . . 8.7 . . . . . . . . . . . . . . . . . . . . . . . . 30995 Golden corn meal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 0.3 3 30996 Yellow granulated corn meal . . . . . . . . . . . . . . . . . . . . . . . . . . 0.35 3 30997 Yellow corn, Illinois . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0.2 5 30998 Yellow hominy feed . . . . . . . . . ._ . . . . . . . . . . . . . . . . . . . . . . . . 0.6 1.5 Vitamin A in the second lot (yellow corn) was determined by the unit method. There seems to be less vitamin A in the meal than in the corn, but the yellow corn meal is well supplied with vitamin A. COMPOSITION OF CORN It is known that the chemical composition of corn varies to some extent according to conditions. By proper selection, corn can be grown which is high or low in protein or high or low in fat. Pendleton (9), fertilization with nitrate of soda had little composition of the grain of corn on Iowa soils. According to efiect on the Delwiche and Totting- ham (2) found the protein in 8 crops of corn grown at Ashland and at VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 35 Madison, Wisconsin, to vary from 9.4 to 13.8 per cent, with an average difference of 0.4 per cent of protein in favor of Ashland. This difference is quite small. The composition of the corn used in the experiments is given in Table 14¢. Averages are given by varieties and by localities in Tables 15 and 16. No significant variation by varieties is observed. The- most sig- nificant variation is in the protein content of the corn grown in different localities. Variation in Protein The significant figures as related to protein are assembled in Table 1'7. The substations are arranged in order according to the average protein content 0f the corn, beginning with the lowest. The figures for 1926, 1927, and 1928 are averaged for several varieties for each substation, but for 1929 there is only one variety, except in case of Temple, for which there are two. The rainfall is also given. The results from the various localities are reasonably concordant, though wide deviations are to be found some years. The corn grown at Nacogdoches in 1928 contained 4 per cent more protein than that grown in other years. The corn grown at College Station decreased regularly in protein content, from 1.2.56 to 9.53 per cent, in the four years. The corn at Denton contained 1 per cent more protein than the average in 1928, and that from Beeville, 1 per cent more in 1928 and 1 per cent less in 1926. In spite of these deviations, the protein content of corn is reasonably constant at a given locality. It is a question how much of the variations shown are due to soil and how much to climatic conditions. It is well known that the composition of cotton seed varies in different sections, containing more protein and less fat in dry sections than in humid sections. It is possible that similar relations exist with reference to the protein of corn and grain sorghums. The correlation coefficient for protein in corn and rainfall (Table 17,) is —- .576 1.072. This is a significant correlation. Variation in Minerals The lime, magnesia, phosphoric acid, and insoluble ash content of some of the samples of corn grown in 1925 is shown in Table 18. The average lime content was slightly lower in the corn from Iowa Park, Beeville, and Angleton, than in that from the other substations. The magnesia was fairly uniform. The phosphoric acid was lower in corn from Beeville and Nacogdoches than in that from the other stations. 36 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION f, .~....-..-...........................ml|lQmflhU>ivn>>mbw Emma amxvm. dofifim wmwzou .E~w.m comtfim =52 . ~ ¢ . | . . . . . . . . . . - . . . . . . . . _ . . . - . - . . NNA wad omAh NNN mmtm mmA: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 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Substation No. 1, Beeville, Texas. 25925 Chisholm . . . . . . . . . . . . . . . . . . . . . .30 .02 .17 .56 . . . . . . . . 25926 Surcropper . . . . . . . . . . . . . . . . . . . . .42 .02 .18 .46 . . . . . . . . 25927 Reese’s Drouth Resister . . . . . . . . . .20 .02 .18 .48 . . . . . . . . 25928 Horton . . . . . . . . . . . . . . . . . . . . . . . . .14 .02 . 18 .45 . . . . . . . . 25929 Oklahoma White Wonder . . . . . . . .16 .02 .17 .47 . . . . . . . . 25930 Hastings Prolific . . . . . . . . . . . . . . . .40 .02 .16 .44 . . . . . . . . 25931 Fentress Strawberry . . . . . . . . . . . . .21 .02 .17 .50 . . . . . . . . 25932 Ferguson Yellow Dent . . . . . . . . . . .20 .02 .17 .50 . . . . . . . . 25933 Thomas . . . . . . . . . . . . . . . . . . . . . . . .18 .02 .17 .43 . . . . . . . . Average (9) . . . . . . . . . . . . . . . . . . 25 .02 .17 .48 . . . . . . . . Substation N0. 11, Nacogdoches, Tex. 25940 Yellow Dent . . . . . . . . . . . . . . . . . . . .22 .02 .16 .48 25941 Brazos White . . . . . . . . . . . . . . . . . . .30 .08 .17 .53 25942 Horton . . . . . . . . . . . . . . . . . . . . . . . . .16 .02 .16 .51 25943 Fentress Strawberry . . . . . . . . . . . . .23 .03 .16 .60 25944 Hastings Prolific . . . . . . . . . . . . . . . .18 .02 .16 .50 25945 Oklahoma White . . . . . . . . . . . . . . . .22 .02 .16 .51 25946 Blue Grain . . . . . . . . . . . . . . . . . . . . .46 .02 .16 .47 25947 Surcropper . . . . . . . . . . . . . . . . . . . . .25 .02 .15 .41 25948 Thomas . . . . . . . . . . . . . . . . . . . . . . . .16 .02 .16 .46 25949 Chisholm . . . . . . . . . . . . . . . . . . . . . . .14 .02 .15 .44 . . . . . . . . 25950 Nacogdoches . . . . . . . . . . . . . . . . . . . .15 .07 .26 .44 . . . . . . . . Average (11) . . . . . . . . . . . . . . .. .22 .03 .17 .49 . . . . . . . . Substation No. 2, Troup, Texas. 25753 Hastings Prolific . . . . . . . . . . . . . . . .05 .03 .17 .66 .0058 25754 Ferguson Yellow Dent . . . . . . . . . . .08 .03 _18 .65 .0065 25755 Surcropper . . . . . . . . . . . . . . . . . . . . .05 .04 .19 .75 .0039 25756 Bloody Butcher . . . . . . . . . . . . . . . . .06 .06 .18 .70 .0047 25757 Fentress Strawberry . . . . . . . . . . . . .02 .02 .17 .66 .0040 25758 Oklahoma White Wonder . . . . . . . . .05 .03 .20 .72 .0028 25759 Chisholm . . . . . . . . . . . . . . . . . . . . . . .03 .02 .18 .74 .0031 25760 Thomas . . . . . . . . . . . . . . . . . . . . . . . .04 .01 .19 .73 .0035 Average (8) . . . . . . . . . . . . . . . .. .05 .03 .18 .70 0043 Substation N0. 6. Denton, Texas. 25793 Oklahoma White Wonder . . . . . . . . .02 ’ .02 .20 . . . . . . . . .0035 25795 Local Squaw . . . . . . . . . . . . . . . . . . . .17 .03 .21 . . . . . . . . .0036 25796 Surcropper . . . . . . . . . . . . . . . . . . . . .10 .03 .20 . . . . . . . . .0075 25797 Bloody Butcher . . . . . . . . . . . . . . . . .09 .04 .21 . . . . . . . . .0074 25798 Chisholm . . . . . . . . . . . . .' . . . . . . . . . .08 .03 .20 . . . . . . . . .0031 25799 Strawberry . . . . . . . . . . . . . . . . . . . . .23 .03 .22 .72 .0036 25800 Brazos White . . . . . . . . . . . . . . . . . . .05 .02 .20 .74 .0050 Average (7) . . . . . . . . . . . . . . . . . . .11 .03 .21 .73 .0048 Substation No. 4. Beaumont, Texas. 25839 Tuxpan . . . . . . . . . . . . . . . . . . . . . . . .12 .05 . 16 .52 . . . . . . . . 25840 Ferguson Yellow Dent . . . . . . . . . . .11 .02 .17 .54 . . . . . . . . 25841 Chisholm . . . . . . . . . . . . . . . . . . . . . . .05 .03 .17 .61 . . . . . . . . 25842 Hastings Prolific . . . . . . . . . . . . . . . .06 .02 . 16 .59 . . . . . . . . 25843 Surcropper . . . . . . . . . . . . . . . . . . . . .05 .02 .17 .60 . . . . . . . . 25844 Fentress Strawberry . . . . . . . . . . . . .06 .02 .15 .67 . . . . . . . . 25845 Thomas . . . . . . . . . . . . . . . . . . . . . . . .04 .02 . 18 .65 . . . . . . . . Average (7) . . . . . . . . . . . . . . . . . .07 .03 . 17 .60 . . . . . . . . 44 BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Table 18.—Mineral composition of corn, 1926—C0ntinued. Insol- _ Phos- No. Where grown and variety uble Lime Mag- phoric Iron Ash nesia Acid Substation N0. 3, Angleton, Texas. 25852 Hastings Prolific . . . . . . . . . . . . . . . .05 .02 .16 .51 . . . . . . . . 25853 Brazos White . . . . . . . . . . . . . . . . . . .06 .02 .16 .51 . . . . . . . . 25854 Oklahoma White Wonder . . . . . . . .07 .02 .16 .51 . . . . . . . . 25855 Tuxpan . . . . . . . . . . . . . . . . . . . . . . . .09 .02 .19 .71 . . . . . . . . 25860 Thomas . . . . . . . . . . . . . . . . . . . . . . . .04 .02 .18 .66 . . . . . . . . Average (5) . . . . . . . . . . . . . . . . . .06 .02 .17 .58 . . . . . . . . ACKNOWLEDGMENT The feeding and care of the rats, and other work in connection with - the assay for vitamin A was chiefly doneby Messrs. R. O. Brooke, H. S. Osgood, and Bay Treichler. Samples of corn were furnished by the superintendents of the various substations. Especial mention should be made of the co-operation of Mr. R. E. Karper, in furnishing samples, and Dr. P. C. Mangelsdorf, in preparing and furnishing corn with definite genetic factors. Other members of the Division of Chemistry and other Divisions of the Agricultural Experiment Station gave aid in various Ways. SUMMARY The diet of man and animals should furnish sufficient energy, pro- teins, vitamins, and minerals such as phosphoric acid, lime, magnesia, soda, chlorine, iron, copper, iodine, and fluorine. Deficiencies in diet may cause retardation of growth in young animals or deficient produc- tion of milk, eggs, etc., or diminished health or vigor, or diseases such as Xerophthalmia, anemia, rickets, pellagra, or goiter. Vitamins are organic compounds which are present in small amounts in foods and are essential to the health of animals. The various vitamins are briefly discussed. Vitamin A is an organic compound essential in very small amounts to animal life. It is found in such foods as green vegetables, butter and corn; it is much more abundant in yellow corn than in white corn. The quantity of vitamin A in foods is measured by experiments on animals, especially rats. Methods of care and breeding of rats are described. Selection of rats resulted in a decided increase in uniformity. Two methods of estimating vitamin A were tested. The unit method (Sherman-Munsell) was found to be much superior to the ration method. Vitamin A was estimated in 20 samples of Ferguson yellow dent corn, grown in 11 different localities and during three seasons, and in 19 samples of other varieties of yellow corn grown in various localities. They contained about the same amount of vitamin A, one gram con- taining 2.5 to 8 units. VARIATIONS IN VITAMIN A AND CHEMICAL COMPOSITION OF CORN 45 Varieties of variegated and of red corn tested (18 samples) were Fentress Strawberry and Bloody Butcher. One gram of Strawberry l corn contained 0.9 to 2.9 units of vitamin A while one gram of the Bloody Butcher contained 1.8 to 2.5 units. Both varieties contained less vitamin A than yellow corn and were more variable. The yellow and the red and the variegated corn grown in 1928 con- tained the most vitamin A, that in 1927 came next, and that grown in 1926 contained the lowest amount. White corn contained so little vitamin A that the amount could be estimated only approximately. It usually contained only one unit in 25 to 35 grams (or more) of corn. A few samples contained a unit in 2 to 5 grams, probably due to crossing with yellow corn. The units of vitamin A in crosses of white and of yellow corn were approximately in proportion to the number of genetic factors present for the yellow endosperm. Samples of corn from mixed pollination containing the yellow genetic factor 1:2 :3 contained vitamin A in the proportions 2.5:5:7 in 1928 and 2:5:8 in 1929. Segregations from a white and a yellow corn contained less vitamin A for the correspond- ing number of genetic factors than the seed from the mixed pollinations, but the ratios were approximately the same. Yellow corn and yellow corn meal are rich in vitamin A. Different varieties of corn showed no significant variation in content of protein, fat, nitrogen-free extract, ash, lime, magnesia, or phosphoric acid. Corn grown in different localities showed variations in lime and phosphoric acid, and especially in the protein content. Corn grown near Beaumont was lowest in protein and that near Bee- ville was highest. The correlation of protein with rainfall was — 0.576 i .072, which is a significant relation. The average lime content was slightly lower in corn grown at Iowa Park, Beeville, and Angleton than at the other localities. The phosphoric acid was slightly lower. in the corn from Beeville and Nacogdoches than from the other places. REFERENCES 1. Bing, Franklin C., and Mendel, Lafayette B., 1929. The Vitamin B and the Vitamin G Requirements of the Albino Bat. J our. Nutrition, 2 :49. 2. Delwiche, E. J. and Tottingham, W. E., 1930. Effect of Climate on Nitrogen Content of Maize, Barley, and Red Clover. Jour. Am. Soc. Agron., 22:681. 3. Dutcher, B. A., 1928. Vitamin B Terminology. Science, 67:206. 4. Eddy, W. H., Gurin, S., and Keresztesy, J., 1930. The Williams- WatermanVitamin B. Jour. of Biol. Chem., 87:729. 5. Evans, H. M., and Lopkovsky, S., 1929. Technic for Determina- tion of the Antineuritic Vitamin B. J our. Nutrition, 2:1. 46 6b. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. BULLETIN NO. 422, TEXAS AGRICULTURAL EXPERIMENT STATION Hauge, S. M., and Trost, J. F., 1928. An Inheritance'Study ~i the Distribution of Vitamin A in Maize. Jour. Biol. Chem ‘ so nor. . Hopkins, F. G., 1921. Effects of Heat and Aeration Upon t“ Fat-soluble Vitamin. Biochem, J ., 14 :7 21 ; Chem. Abstrac if; 15, 1556. 1 Mangelsdorf, P. C., and Fraps, G. S., 1931. A Direct Quanti tative Relationship between Vitamin A in Corn and the Numb of Genes for Yellow Pigmentation. Science, 73 (N. S.): 241. Meyer, C. R., and Hetler, R. A., 1929. The Distribution 0 Vitamin A in Some Corn-Milling Products. J our. Agr. Research; 39 z767. ' Nelson, E. M., and Jones, D. B., 1928. Observations Bearing on the Determination of Vitamin A. Jour. of Biol. Chem., 80 :215.j_ Pendleton, R. A., 1930. Sodium Nitrate as a Fertilizer for Corn; on Iowa Soils. Jour. of theAm. Soc. Agron, 22:673. Rocke, C., and Hetler, R. A., 1928. The Content and Distribution; of Vitamin A in the Milling Products of Yellow Corn. U. S.f Dept. Agri, Circular 84, p. 17. f p Russell, W. C., 1930. The Vitamin A Content of Yellow and‘. White-Capped Yellow Dent Corn. Jour. Nutrition, 2:265. Sherman, H. C., and Burtis, M. P., 1928. Factors Affecting the Accuracy of the Quantitative Determination of Vitamin A. J our. . Biol. Chem., 78:671. ' ; Sherman, H. C., and Munsell, H. E., 1925. The Quantitative De- termination of Vitamin A. J our. Am. Chem. Soc., 47 :1639. '_ Steenbock, H., and Boutwell, P. W., 1920. Fat-Soluble Vitamins“. III. The Comparative Nutritive Value of White and Yellow l_ Maizes. Jour. Biol. Chem., 41:81. Steenbock, H., and Coward, K. H., 1927. Fat-Soluble Vitamins. 5 XXVII. The Quantitative Determination o1": Vitamin A. Jour. Biol. Chem." 72:765-799. Smith, M. C., 1930. A Quantitative Comparison of the Vitamin f, A Content of Yellow Corn and the Grain Sorghums Hegari and Yellow Milo. Jour. of Agri. Res, 40:1147. Smith, M. 0., 1930. f{_ Corn and the Grain Sorghums Hegari and Yellow Milo. J our. of Agr. Res., 40:1129. A Smith, S. L.,>1928. Science, 67:494. Smith, S. L., 1929. Vitamins in Food Materials. Agri., Circular No. 84. Vitamin B. A Question of Nomenclature. The Comparative Nutritive Value of Yellow A U. Dept. of .