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The Isolation and Identification of Glucose from the Albumen of the Hen’s Egg1
546
J. H. STRAIN AND A. S. JOHNSON
est in October; the seasonal pattern varied somewhat between different strains. A statistically significant negative regression of blood spot incidence on clutch position was observed. REFERENCES
The Isolation and Identification of Glucose from the Albumen of the Hen's Egg A L L E N B . SCHLESINGER
The Departments of Biology and Agricultural Biochemistry, The Creighton University, Omaha, Nebraska, and The University of Minnesota, St. Paul, Minnesota1 (Received for publication November 22, 1956)
ANALYSES of the free carbohydrate - t * - components of the albumen can be traced to the isolation (Winckler, 1846) of a substance identified as lactose. Subsequent studies failed to verify the report 1
The provision of facilities and guidance by Dr. Fred Smith of the Department of Agricultural Biochemistry of the University of Minnesota is gratefully acknowledged.
but led to the assertion by Budge (1848) and Aldridge (1849) that glucose was a constituent of egg white. Needham (1931) lists eight investigations of glucose from this source before the end of the nineteenth century. Many quantitative determinations followed and Romanoff and Romanoff (1949) gives a value of 0.4% free glucose in the albumen in their comprehensive study.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
Carver, J. S., and W. Henderson, 1948. The effect of rutin and ascorbic acid and of alfalfa on blood and meat spots in hen's eggs. Poultry Sci. 27:656. Goodman, B. L., and G. F. Godfrey, 1955. Genetic, phenotypic and environmental correlations between some egg quality traits and egg production and hatchability. Poultry Sci. 34: 1197. Gowe, R. S., and A. S. Johnson, 1956. The performance of a control strain of S. C. White Leghorn stock over four generations on test at several locations. Poultry Sci. 35: 1146. Gowe, R. S., and W. J. Wakely, 1954. Environment and poultry breeding problems. 1. The influence of several environments on the egg production and viability of different genotypes. Poultry Sci. 33:691-703. Jeffrey, F. P., 1941. Changes in pullet year albumen index as affected by age of bird. Poultry Sci. 20: 298-301. Jeffrey, F. P., and J. Pino, 1943. The effects of heredity and of certain environmental factors on the incidence of blood spots in chicken eggs. Poultry Sci. 22:230-234.
Jeffrey, F. P., and C. E. Walker, 1950. The relationship between egg shell color and incidence of colored meat spots. Poultry Sci. 29: 244-247. Johnson, A. S., 1956. Genetic and seasonal variation in incidence of blood and meat spots in chicken eggs. Can. J. Agr. Sci. 36: 390-393. Johnson, A. S., and E. S. Merritt, 1955. Heritability of albumen height and specific gravity of eggs from White Leghorns and Barred Rocks and the correlations of these traits with egg production. Poultry Sci. 34: 578-587. King, S. C , and G. O. Hall, 1955. Egg quality studies at the New York random sample test. Poultry Sci. 34: 799-809. King, S. C , and C. R. Henderson, 1954. Heritability studies of egg production in the domestic fowl. Poultry Sci. 33: 155-169. Nalbandov, A. V., and L. E. Card, 1944. The problem of blood clots and meat spots in chicken eggs. Poultry Sci. 23:170-180. Sauter, E. A., W. J. Stadelman and J. S. Carver, 1952. Factors affecting the incidence of blood spots and their detection in hens' eggs. Poultry Sci. 31: 1042-1049. Skaller, F., 1954. Studies on the assessment of egg production in poultry breeding investigations. 2. The correction of hatching date for early selection based on part-annual egg production records. Poultry Sci. 33:316-321. Snedecor, G. W., 1946. Statistical Methods Applied to Experiments in Agriculture and Biology. 485 pp. Iowa State College Press, Ames, Iowa.
GLUCOSE IN EGG ALBUMEN
An osazone melting at 203-204°C. was prepared from the non-coagulable (with alcohol) portion of the albumen by Salkowski (1893). There is reason to suspect that this preparation involved both glucose and fructose, for Morner (1912), in a very extensive report, noted that the calculated value for glucose obtained from polarimetric data was slightly lower than that obtained using a Fehling titration. Morner suggested the presence of a laevorotatory component and/or reducing components other than glucose. He indicated conclusively that the reducing capacity of the albumen extract was quantitatively destroyed following treatment with yeast but was unable to demonstrate the presence of a keto compound. As will be developed below, however, it appears that low concentrations of fructose exist in some, if not all, egg whites. Lowering the reducing sugar content of the albumen plays a significant role in the processing of eggs for commercial dehydration without darkening. A variety of processes have been developed to accomplish this purpose. Some have little, if any, specificity with regard to the substance (s)
removed, e.g., by dialysis (Moriarity et al., 1952), fermentation (Stewart et al., 1941). Others however, appear to be highly selective in the removal of glucose; Scott (1953) utilizes preparations of glucose oxidase in conjunction with hydrogen peroxide to effect the conversion of glucose to glucuronic acid. The presence of glucose in eggs has been determined by chromatographic analysis (McFarren et al, 1951). This paper is concerned with the isolation of Z)-glucose and its characterization as a crystalline derivative. EXPERIMENTAL
The eggs used were provided by the Poultry Division of the Institute of Agriculture of the University of Minnesota. These White Leghorn eggs were obtained three hours after laying and were processed immediately. The albumen from 10 eggs was collected. No effort was made to separate the various layers nor was particular emphasis placed upon obtaining the chalaziferous layer. The albumen from the individual eggs was mixed and the mixture dissolved in an equal volume of distilled water. This preparation was dialyzed in the cold for 6 hours against 5 volumes of distilled water. The dialysate was concentrated to a syrup by distillation in vacuo at 50° C. A small portion of this residue gave a strong Fehling test. The syrup was dissolved in 10 ml. distilled water and the solution passed through columns of Amberlite IR-120 and Duolite A4 for cation and anion removal respectively. CHROMATOGRAPHIC ANALYSIS
The analysis was carried out using Whatman No. 1 paper at a temperature of approximately 25° C. using the descending technique. Identification was by com-
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
A variety of methods have been employed in the determinations referred to. These include measurement of evolved CO2 resulting from fermentation (Lehmann, 1853), polarimetry of the dialyzable albumen components (Salkowski, 1893), and a variety of volumetric and colorimetric procedures which utilize the reducing capacity of glucose. Most of the more recent studies have utilized the Fehling, Benedict, Hagedorn-Jensen, or Shaffer-Hartmann techniques (Morner, 1912; Vladimirov et al., 1926; Needham, 1927; Donhoffer, 1933; and Hopkins et al., 1948). Quantitative studies have generally relied upon reducing capacity of albumen extracts prior to, and subsequent to, fermentation.
547
548
A. B . SCHLESINGER
parison with standard glucose solutions run integrally. Solvents used, duration of runs, and visualizing reagents are indicated.
ture values, m.p. 184°C, [a] D 20 -202° in pyridine (C, 1.0). A 149 gm. sample of albumen extracted quantitatively yielded .27% glucose.
Solvent
In all cases the unknown corresponded to glucose. In addition a second reducing component corresponding to fructose was found to be present in small amounts. ISOLATION AND CHARACTERIZATION OF D-GLUCOSE The entire sample was placed on Whatman No. 1 filter paper according to the method of Dubois et al. (1956), standards of glucose and fructose being placed in the marginal strips on the starting line. The chromatogram was irrigated for 18 hours using the pyridine; ethyl acetate; water solvent. The marginal strips were cut off and the sugars located with Tollens ammoniacal silver nitrate reagent. The chromatogram was reassembled and the glucose and fructose bearing zones were cut out from the central unsprayed portions of the chromatogram. Separate elutions in a total of 20 ml. of distilled water were filtered and evaporated to constant weight in vacuo at 50°C. The samples of glucose and fructose weighed 10 and 0.9 mg. respectively. The glucose component upon treatment with p-nitroaniline and a trace of acid according to the method of Weygand et al. (1951), afforded A^-p-nitrophenyl Dglucosamine, m.p. and mixed m.p. 183°C, [a] D 25 -203° in pyridine (C, 0.7); litera-
Detection reagent
48 18 36 30 18
AgN0 3 -NH 4 OH AgN0 3 -NH 4 OH AgN0 3 -NH 4 OH ^-anisidine |>-anisidine
SUMMARY
Glucose from the dialysate of hen's egg albumen was isolated by paper partition chromatography and identified by the preparation and characterization of the crystalline p-nitroanilide. REFERENCES Aldridge, J., 1849. Discovery of the constant presence of grape sugar in the white of eggs. Medical Times (London), 19: 437. Budge, O., 1848. Zucker in Huhnereiweiss. Ann. Chem. u. Pharm. 64: 127-128. Donhoffer, C , 1933. CVII. Studies on the carbohydrate metabolism of the chick embryo. Biochem. J. 27:806-817. Dubois, M., K. A. Gillis, J. K. Hamilton, P. A. Rebers and F. Smith, 1956. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350-356. Hopkins, J. W., and R. M. Trevoy, 1948. Liquid and frozen egg. IV. Reproducibility of measurements of reducing sugar in frozen egg. Can. J. Research, 26F: 221-227. Lehmann, C. G., 1853. Physiological Chemistry, Vol. 2, Harrison, London. McFarren, E. F., K. Brand and H. R. Rutkowski, 1951. Quantitative determination of sugars on filter paper chromatograms by direct photometry. Anal. Chem. 23: 1146-1149. Moriarity, J. H., and R. P. Joslin, 1952. Egg preservation. Official Gaz. U. S. Pat. Off. 656: 893-894. Morner, C. T., 1912. Uber ovomucoid und zucker in dem weissen der vogeleier; eine systematische untersuchung. Zeitschr. f. Physiol. Chem. 80: 431-473. Needham, J., 1927. The energy sources in onto-
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
1-Butanol; ethanol; water (4:1:5) (Partridge et al, 1948) Pyridine; ethyl acetate; water (1:2.5:3.5) (McFarren et al., 1951) 1-Butanol; acetic acid; water (2:1:1) (Partridge et al., 1948) Phenol saturated with water (Partridge et al., 1948) Pyridine; ethyl acetate; water (1:2.5:3.5) (McFarren et al., 1951)
Hours
549
GLUCOSE IN EGG ALBUMEN
Scott, D., 1953. Identification of enzyme desugarized egg solids. J. Agr. Food Chem. 1:1109-1111. Stewart, G. F., and R. W. Kline, 1941. Dried egg albumin. I. Solubility and color denaturation. Proc. Inst. Food Technol. 1941: 48-56. Vladimirov, G. E., and A. A. Schmidt, 1926. Beitrage zur embryochemie und embryophysiologie. Biochem. Zeitschr. 177:298-303. Weygand, F., W. Perkow and P. Kuhner, 1951. ^-Glycosides VII. ^-nitroanaline glycosides and a synthesis of gentiobiose from ^>-nitroanaline iV-glucoside. Chem. Ber. 84: 594-602. Winckler, F. L., 1846. Zucker in vogeleier. Repert. f. d. Pharmazie 42: 46-49.
The Use of Vitamin A Protected by D P P D (Diphenyl-pPhenylene-Diamine) for the Growth of Chickens I . ASCARELLI Faculty of Agriculture, Hebrew University, Rehovot, Israel (Received for publication November 22, 1956)
T
HE use of fish oils in poultry nutrition, quite widespread in Israel up to this time, brought about an irrational wastage of vitamin A because of losses which may take place during storage of the feed and probably also during passage through the intestinal tract of the bird. To avoid these losses many manufacturers now add DPPD to their mashes. Matterson et al. (1955a) showed DPPD to be nontoxic to poultry. In another effort to avoid the abovementioned losses, many poultry men have changed over to the use of dry stabilized (coated) vitamin A supplements. In order to ascertain whether the coating process, besides increasing the stability of vitamin A in vitro also affects the utilization of this vitamin by the bird, a nutrition experiment was carried out with chickens. In this experiment fish oil and dry coated vitamin A concentrate were used as vitamin A sources, both in presence and absence of DPPD.
EXPERIMENTAL
One-day old S. C. White Leghorn chickens were grown in wire-floored battery brooders. They were weighed individually when 1, 7, and 14 days old, and divided into groups of 50 on the basis of weight at 7 days. At 4 weeks of age the chickens were divided into sub-groups by sex, and at 5 weeks were transferred to an outdoor growing house. The percent composition of the diet was: ground barley— 15, ground white corn—25.25, wheat bran—20, peanut meal—20, dried skimmilk—5, South African fish meal (58% protein)—7.5, alfalfa hay meal (carotene free)—4, ground oyster shell—2, dibasic calcium phosphate—0.75, manganized salt •—0.5. The diet was supplemented with 100 I.U./IOO g. vitamin D3, riboflavin 3 g./ton, calcium pantothenate 11 g./ton, vitamin Bi2 5 mg./ton. The diet contained by analysis 20.3% protein, 6.3% fiber, 2.4% calcium and 1.1% phosphorus. The first two weeks served as a depletion
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
genesis. V. The carbohydrate metabolism of the developing avian egg. Brit. J. Exp. Biol. 5: 6-42. Needham, J., 1931. Chemical Embryology, Vol. 1, Cambridge. Partridge, S. M., and R. G. Westall, 1948. Filter paper partition chromatography of sugars. I. General description and application to the qualitative analysis of sugars in apple juice, egg white, and fetal blood of sheep. Biochem. J. 42:238-250. Romanoff, A. R., and A. J. Romanoff, 1949. The Avian Egg. Wiley, N. Y. Salkowski, E., 1893. Zur chemie des albumens des huhnereies. Centrblt. f. d. med. Wiss. 31: 513515.
J. H. STRAIN AND A. S. JOHNSON
est in October; the seasonal pattern varied somewhat between different strains. A statistically significant negative regression of blood spot incidence on clutch position was observed. REFERENCES
The Isolation and Identification of Glucose from the Albumen of the Hen's Egg A L L E N B . SCHLESINGER
The Departments of Biology and Agricultural Biochemistry, The Creighton University, Omaha, Nebraska, and The University of Minnesota, St. Paul, Minnesota1 (Received for publication November 22, 1956)
ANALYSES of the free carbohydrate - t * - components of the albumen can be traced to the isolation (Winckler, 1846) of a substance identified as lactose. Subsequent studies failed to verify the report 1
The provision of facilities and guidance by Dr. Fred Smith of the Department of Agricultural Biochemistry of the University of Minnesota is gratefully acknowledged.
but led to the assertion by Budge (1848) and Aldridge (1849) that glucose was a constituent of egg white. Needham (1931) lists eight investigations of glucose from this source before the end of the nineteenth century. Many quantitative determinations followed and Romanoff and Romanoff (1949) gives a value of 0.4% free glucose in the albumen in their comprehensive study.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
Carver, J. S., and W. Henderson, 1948. The effect of rutin and ascorbic acid and of alfalfa on blood and meat spots in hen's eggs. Poultry Sci. 27:656. Goodman, B. L., and G. F. Godfrey, 1955. Genetic, phenotypic and environmental correlations between some egg quality traits and egg production and hatchability. Poultry Sci. 34: 1197. Gowe, R. S., and A. S. Johnson, 1956. The performance of a control strain of S. C. White Leghorn stock over four generations on test at several locations. Poultry Sci. 35: 1146. Gowe, R. S., and W. J. Wakely, 1954. Environment and poultry breeding problems. 1. The influence of several environments on the egg production and viability of different genotypes. Poultry Sci. 33:691-703. Jeffrey, F. P., 1941. Changes in pullet year albumen index as affected by age of bird. Poultry Sci. 20: 298-301. Jeffrey, F. P., and J. Pino, 1943. The effects of heredity and of certain environmental factors on the incidence of blood spots in chicken eggs. Poultry Sci. 22:230-234.
Jeffrey, F. P., and C. E. Walker, 1950. The relationship between egg shell color and incidence of colored meat spots. Poultry Sci. 29: 244-247. Johnson, A. S., 1956. Genetic and seasonal variation in incidence of blood and meat spots in chicken eggs. Can. J. Agr. Sci. 36: 390-393. Johnson, A. S., and E. S. Merritt, 1955. Heritability of albumen height and specific gravity of eggs from White Leghorns and Barred Rocks and the correlations of these traits with egg production. Poultry Sci. 34: 578-587. King, S. C , and G. O. Hall, 1955. Egg quality studies at the New York random sample test. Poultry Sci. 34: 799-809. King, S. C , and C. R. Henderson, 1954. Heritability studies of egg production in the domestic fowl. Poultry Sci. 33: 155-169. Nalbandov, A. V., and L. E. Card, 1944. The problem of blood clots and meat spots in chicken eggs. Poultry Sci. 23:170-180. Sauter, E. A., W. J. Stadelman and J. S. Carver, 1952. Factors affecting the incidence of blood spots and their detection in hens' eggs. Poultry Sci. 31: 1042-1049. Skaller, F., 1954. Studies on the assessment of egg production in poultry breeding investigations. 2. The correction of hatching date for early selection based on part-annual egg production records. Poultry Sci. 33:316-321. Snedecor, G. W., 1946. Statistical Methods Applied to Experiments in Agriculture and Biology. 485 pp. Iowa State College Press, Ames, Iowa.
GLUCOSE IN EGG ALBUMEN
An osazone melting at 203-204°C. was prepared from the non-coagulable (with alcohol) portion of the albumen by Salkowski (1893). There is reason to suspect that this preparation involved both glucose and fructose, for Morner (1912), in a very extensive report, noted that the calculated value for glucose obtained from polarimetric data was slightly lower than that obtained using a Fehling titration. Morner suggested the presence of a laevorotatory component and/or reducing components other than glucose. He indicated conclusively that the reducing capacity of the albumen extract was quantitatively destroyed following treatment with yeast but was unable to demonstrate the presence of a keto compound. As will be developed below, however, it appears that low concentrations of fructose exist in some, if not all, egg whites. Lowering the reducing sugar content of the albumen plays a significant role in the processing of eggs for commercial dehydration without darkening. A variety of processes have been developed to accomplish this purpose. Some have little, if any, specificity with regard to the substance (s)
removed, e.g., by dialysis (Moriarity et al., 1952), fermentation (Stewart et al., 1941). Others however, appear to be highly selective in the removal of glucose; Scott (1953) utilizes preparations of glucose oxidase in conjunction with hydrogen peroxide to effect the conversion of glucose to glucuronic acid. The presence of glucose in eggs has been determined by chromatographic analysis (McFarren et al, 1951). This paper is concerned with the isolation of Z)-glucose and its characterization as a crystalline derivative. EXPERIMENTAL
The eggs used were provided by the Poultry Division of the Institute of Agriculture of the University of Minnesota. These White Leghorn eggs were obtained three hours after laying and were processed immediately. The albumen from 10 eggs was collected. No effort was made to separate the various layers nor was particular emphasis placed upon obtaining the chalaziferous layer. The albumen from the individual eggs was mixed and the mixture dissolved in an equal volume of distilled water. This preparation was dialyzed in the cold for 6 hours against 5 volumes of distilled water. The dialysate was concentrated to a syrup by distillation in vacuo at 50° C. A small portion of this residue gave a strong Fehling test. The syrup was dissolved in 10 ml. distilled water and the solution passed through columns of Amberlite IR-120 and Duolite A4 for cation and anion removal respectively. CHROMATOGRAPHIC ANALYSIS
The analysis was carried out using Whatman No. 1 paper at a temperature of approximately 25° C. using the descending technique. Identification was by com-
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
A variety of methods have been employed in the determinations referred to. These include measurement of evolved CO2 resulting from fermentation (Lehmann, 1853), polarimetry of the dialyzable albumen components (Salkowski, 1893), and a variety of volumetric and colorimetric procedures which utilize the reducing capacity of glucose. Most of the more recent studies have utilized the Fehling, Benedict, Hagedorn-Jensen, or Shaffer-Hartmann techniques (Morner, 1912; Vladimirov et al., 1926; Needham, 1927; Donhoffer, 1933; and Hopkins et al., 1948). Quantitative studies have generally relied upon reducing capacity of albumen extracts prior to, and subsequent to, fermentation.
547
548
A. B . SCHLESINGER
parison with standard glucose solutions run integrally. Solvents used, duration of runs, and visualizing reagents are indicated.
ture values, m.p. 184°C, [a] D 20 -202° in pyridine (C, 1.0). A 149 gm. sample of albumen extracted quantitatively yielded .27% glucose.
Solvent
In all cases the unknown corresponded to glucose. In addition a second reducing component corresponding to fructose was found to be present in small amounts. ISOLATION AND CHARACTERIZATION OF D-GLUCOSE The entire sample was placed on Whatman No. 1 filter paper according to the method of Dubois et al. (1956), standards of glucose and fructose being placed in the marginal strips on the starting line. The chromatogram was irrigated for 18 hours using the pyridine; ethyl acetate; water solvent. The marginal strips were cut off and the sugars located with Tollens ammoniacal silver nitrate reagent. The chromatogram was reassembled and the glucose and fructose bearing zones were cut out from the central unsprayed portions of the chromatogram. Separate elutions in a total of 20 ml. of distilled water were filtered and evaporated to constant weight in vacuo at 50°C. The samples of glucose and fructose weighed 10 and 0.9 mg. respectively. The glucose component upon treatment with p-nitroaniline and a trace of acid according to the method of Weygand et al. (1951), afforded A^-p-nitrophenyl Dglucosamine, m.p. and mixed m.p. 183°C, [a] D 25 -203° in pyridine (C, 0.7); litera-
Detection reagent
48 18 36 30 18
AgN0 3 -NH 4 OH AgN0 3 -NH 4 OH AgN0 3 -NH 4 OH ^-anisidine |>-anisidine
SUMMARY
Glucose from the dialysate of hen's egg albumen was isolated by paper partition chromatography and identified by the preparation and characterization of the crystalline p-nitroanilide. REFERENCES Aldridge, J., 1849. Discovery of the constant presence of grape sugar in the white of eggs. Medical Times (London), 19: 437. Budge, O., 1848. Zucker in Huhnereiweiss. Ann. Chem. u. Pharm. 64: 127-128. Donhoffer, C , 1933. CVII. Studies on the carbohydrate metabolism of the chick embryo. Biochem. J. 27:806-817. Dubois, M., K. A. Gillis, J. K. Hamilton, P. A. Rebers and F. Smith, 1956. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350-356. Hopkins, J. W., and R. M. Trevoy, 1948. Liquid and frozen egg. IV. Reproducibility of measurements of reducing sugar in frozen egg. Can. J. Research, 26F: 221-227. Lehmann, C. G., 1853. Physiological Chemistry, Vol. 2, Harrison, London. McFarren, E. F., K. Brand and H. R. Rutkowski, 1951. Quantitative determination of sugars on filter paper chromatograms by direct photometry. Anal. Chem. 23: 1146-1149. Moriarity, J. H., and R. P. Joslin, 1952. Egg preservation. Official Gaz. U. S. Pat. Off. 656: 893-894. Morner, C. T., 1912. Uber ovomucoid und zucker in dem weissen der vogeleier; eine systematische untersuchung. Zeitschr. f. Physiol. Chem. 80: 431-473. Needham, J., 1927. The energy sources in onto-
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
1-Butanol; ethanol; water (4:1:5) (Partridge et al, 1948) Pyridine; ethyl acetate; water (1:2.5:3.5) (McFarren et al., 1951) 1-Butanol; acetic acid; water (2:1:1) (Partridge et al., 1948) Phenol saturated with water (Partridge et al., 1948) Pyridine; ethyl acetate; water (1:2.5:3.5) (McFarren et al., 1951)
Hours
549
GLUCOSE IN EGG ALBUMEN
Scott, D., 1953. Identification of enzyme desugarized egg solids. J. Agr. Food Chem. 1:1109-1111. Stewart, G. F., and R. W. Kline, 1941. Dried egg albumin. I. Solubility and color denaturation. Proc. Inst. Food Technol. 1941: 48-56. Vladimirov, G. E., and A. A. Schmidt, 1926. Beitrage zur embryochemie und embryophysiologie. Biochem. Zeitschr. 177:298-303. Weygand, F., W. Perkow and P. Kuhner, 1951. ^-Glycosides VII. ^-nitroanaline glycosides and a synthesis of gentiobiose from ^>-nitroanaline iV-glucoside. Chem. Ber. 84: 594-602. Winckler, F. L., 1846. Zucker in vogeleier. Repert. f. d. Pharmazie 42: 46-49.
The Use of Vitamin A Protected by D P P D (Diphenyl-pPhenylene-Diamine) for the Growth of Chickens I . ASCARELLI Faculty of Agriculture, Hebrew University, Rehovot, Israel (Received for publication November 22, 1956)
T
HE use of fish oils in poultry nutrition, quite widespread in Israel up to this time, brought about an irrational wastage of vitamin A because of losses which may take place during storage of the feed and probably also during passage through the intestinal tract of the bird. To avoid these losses many manufacturers now add DPPD to their mashes. Matterson et al. (1955a) showed DPPD to be nontoxic to poultry. In another effort to avoid the abovementioned losses, many poultry men have changed over to the use of dry stabilized (coated) vitamin A supplements. In order to ascertain whether the coating process, besides increasing the stability of vitamin A in vitro also affects the utilization of this vitamin by the bird, a nutrition experiment was carried out with chickens. In this experiment fish oil and dry coated vitamin A concentrate were used as vitamin A sources, both in presence and absence of DPPD.
EXPERIMENTAL
One-day old S. C. White Leghorn chickens were grown in wire-floored battery brooders. They were weighed individually when 1, 7, and 14 days old, and divided into groups of 50 on the basis of weight at 7 days. At 4 weeks of age the chickens were divided into sub-groups by sex, and at 5 weeks were transferred to an outdoor growing house. The percent composition of the diet was: ground barley— 15, ground white corn—25.25, wheat bran—20, peanut meal—20, dried skimmilk—5, South African fish meal (58% protein)—7.5, alfalfa hay meal (carotene free)—4, ground oyster shell—2, dibasic calcium phosphate—0.75, manganized salt •—0.5. The diet was supplemented with 100 I.U./IOO g. vitamin D3, riboflavin 3 g./ton, calcium pantothenate 11 g./ton, vitamin Bi2 5 mg./ton. The diet contained by analysis 20.3% protein, 6.3% fiber, 2.4% calcium and 1.1% phosphorus. The first two weeks served as a depletion
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 9, 2015
genesis. V. The carbohydrate metabolism of the developing avian egg. Brit. J. Exp. Biol. 5: 6-42. Needham, J., 1931. Chemical Embryology, Vol. 1, Cambridge. Partridge, S. M., and R. G. Westall, 1948. Filter paper partition chromatography of sugars. I. General description and application to the qualitative analysis of sugars in apple juice, egg white, and fetal blood of sheep. Biochem. J. 42:238-250. Romanoff, A. R., and A. J. Romanoff, 1949. The Avian Egg. Wiley, N. Y. Salkowski, E., 1893. Zur chemie des albumens des huhnereies. Centrblt. f. d. med. Wiss. 31: 513515.