121. Osborne and MENDEL: The relation of growth to the chemical constituents of the diet; J. Biol. Chem., 15, 311, 1913.

122. HART and MCCOLLUM: Influence on growth of rations restricted to the corn or wheat-grain; J. Biol. Chem., 19, 373, 1914. 123. DESANI: Untersuchungen über die Genese des Cholesterins;

Chem. Zentr., 2, 1764, 1913.

124. MCCOLLUM and DAVIS: Further observations on the physiological properties of the lipins of the egg-yolk; Proc. Soc. Exp. Biol. Med., 11, 101, 1914.

125. LANDER: On the cholesterol content of the tissues of growing rats when under various diets; Biochem. J., 9, 78, 1915.

126. MACARThur and LucKETT: Lipins in nutrition; J. Biol. Chem., 20, 161, 1915.

127. MCCOLLUM and Davis: The necessity of certain lipins in the diet

during growth; Ibid., 15, 167, 1913.

128. OSBORNE and MENDEL: The relationship of growth to the chemical constituents of the diet; Ibid., 15, 311, 1913.

129. —: The influence of butter-fat on growth; Ibid., 16, 423, 1913.

130. FUNK and MACALLUM: Die chemischen Determinanten des

Wachstums; Z. physiol. Chem., 92, 13, 1914.

131. MCCOLLUM and DAVIS: Observations on the isolation of the substance in butter-fat which exerts a stimulating influence on growth; J. Biol. Chem., 19, 245, 1914.

132. Osborne and WAKEMAN: Does butter-fat contain nitrogen and phosphorus?; Ibid., 21, 91, 1915.

133. ARON: Untersuchungen über die Beeinflussung des Wachstums durch Ernährung; Berliner klin. Woch., No. 21, 1914.

134. OSBORNE and MENDEL: The suppression of growth and the process of growth; J. Biol. Chem., 18, 95, 1914.

135. MCCOLLUM and Davis: Nutrition with purified food substances;

Ibid., 20, 641, 1915.

136. OSBORNE and MENDEL: The influence of cod-liver oil and some other fats on growth; Ibid., 17, 401, 1914.

137.

138.

139.

: Further observations on the influence of natural fats upon growth; Ibid., 20, 379, 1915.

1914.

: Some problems of growth; Ibid., 17, 2, 1914.

: Amino-acids in nutrition and growth; Ibid., 17, 325,

: The comparative nutritive values of certain proteins in growth, and the problem of protein minimum; Ibid., 20, 351, 1915.

141. MCCOLLUM: The value of the proteins of the cereal grains and of milk for growth of the pig, and the influence of the plane of protein intake on growth; Ibid., 19, 329, 1914.

142. STREET: The feeding value of sanatogen compared with commercial casein with respect to maintenance and growth; J. Am. Med. Assoc., 63, 1831, 1914.

143. HEKTOEN: The formation of antibodies in rats fed on pure vegetable proteins. (Osborne-Mendel stunting diet); J. Infect. Dis., 15, 278, 1914.

144. FUNK: Studien über das Wachstum. Das Wachstum auf vitaminhaltiger und vitaminfreier Nahrung; Z. physiol. Chem., 88, 352, 1913.

145.

: Studies on growth: The influence of diet on growth, normal and malignant; Lancet, Jan. 10th, 1914. 146. MCCOLLUM and Davis: The influence of certain vegetable fats

on growth; J. Biol. Chem., 21, 179, 1915.

147. BOTTOMLEY: Some accessory factors in plant growth and nutri

148.

tion; Proc. Roy. Soc. (B), 88, 237, 1914.

: The significance of certain food substances for plant growth; Ann. Botany, 28, 531, 1914.

Tumors

149. Rous: The influence of diet on transplanted and spontaneous mouse tumors; J. Exp. Med., 20, 433, 1914.

150. CENTANNI: La dieta aviride per lo sviluppo dei tumori sperimentali; Tumori, 2, 466, 1914.

151. VAN ALstyne and BeEBE: The effect of non-carbohydrate diet upon the growth of sarcoma in rats; J. Med. Res., 29, 219, 1913.

152. BENEDICT and LEWIS: The influence of induced diabetes on malignant tumors (including a report of a case of human phlorhizin glycosuria); Proc. Soc. Exp. Biol. Med., 11, 134, 1914.

153. RONDONI: Esperienze sui tumori. VIII; Riunc. Soc. Ital. di Patol. Pisa, Mar., 1913.

154. NASSETTI: Innesti eterogenei dei tumori; Tumori, 3, Nov.-Dec., 1913.

155. MURPHY: Transplantation of tissues to the embryo of foreign species; J. Exp. Med., 17, 482, 1913.

156. MURPHY and MORTON: The lymphocyte in natural and induced resistance to transplanted cancer, J. Exp. Med., 22, 204, 1915. 157. FUNK: The transplantation of tumors to foreign species; J. Exp. Med., 21, 571, 1915.

158. V. GRAFF: Der Einfluss der Schwangerschaft auf das Wachstum maligner Tumoren; Wiener klin. Wochschr., 27, 7, 1914. 159. ROBERTSON and BURNETT: Influence of the anterior lobe of the pituitary body upon the growth of carcinomata; J. Exp. Med., 21, 280, 1915.

160. FUNK: Serum diagnosis of Rous's chicken sarcoma, based on chemical methods; BIOCHEM. BULL., 4, 24, 1915.

General nutrition

161. MELOCCHI: Nuovi orizzonti sull'alimentazione dell'uomo, Napoli,

1914.

162, FRIEDENTHAL: Ueber Säuglingsernährung nach physiologischen Grundsätzen mit Friedenthal'schen Kindermilch und Gemüsepulvern; Berl. klin. Wochschr., No. 16, 1914.

163. STERNBERG: Diät und diätetische Behandlung vom Standpunkte der Vitaminlehre und vom Standpunkte der Lehre der diätetischen Küche; Arch. Verdauungs-krankh., 20, 200,

1914.

164. KUNERT: Unsere heutige falsche Ernährung; Breslau, 1914. 165. Hüssy: Zur klinischen Bedeutung der Vitamine; Münch. med.

Wochschr., 61, 18, 1914.

166. REACH: Studien über die Nebenwirkungen der Nahrungstoffe;

Ber. Wien. Akad., Abt. III, 1, 1913.

167. PEISER: Fett in Kindernahrung; Berl. klin. Wochschr., 51, 1065,

1914.

168. MORRISON: Some of the dangers of too greatly restricted diet in typhoid; Kentucky Med. J., 12, Nov. 15th, 1914.

169. THOMAS: Beziehungen der chronischen Unterernährung zur Infektion und zu klinischen Zeichen der verminderten Immunität; Z. Kinderh., July, 1914.

170. RÉNON: La tuberculose et les vitamines; Soc. de therapeut. de

Paris, 24th June, 1914.

BIOCHEMISTRY OF COD-LIVER OIL.

(Preliminary note)

CASIMIR FUNK

INTRODUCTION. It is very surprising that the chemistry and physiology of cod-liver oil have been neglected to such a degree that the author was able to find only a few papers dealing with the subject.

In 1888, Gautier and Mourgues (1) showed that even the refined yellow oil contains a small quantity of organic bases (which were then considered to be ptomaines or alkaloids). Their technic was as follows: 100 k. of yellow cod-liver oil were extracted with the same volume of 33 percent alcohol, to which 4 gm. of oxalic acid per 1. were added. The aqueous-alcoholic extracts were saturated with calcium hydroxid, filtered and evaporated in vacuo, at 45° C. Toward the end of the distillation, precipitated calcium carbonate and calcium hydroxid were added, the mixture evaporated to dryness, and the residue extracted with 90 percent alcohol. From this alcoholic extract the alcohol was removed in vacuo, water and strong caustic potash were added to the residue, and the alkaline mixture was extracted with ether. To the ethereal extract, a sol. of oxalic acid in ether was added; this precipitated the bases as oxalates. The yield was 52-65 gm. of oxalates from 100 k. of cod-liver oil. The oxalates were dissolved in dil. caustic potash sol., and the free bases separated, as an oil, on the surface of the liquid The oil was removed and dried over freshly calcinated potash. In this way 0.35-0.5 gm. of dry substance was obtained.

Subjected to fractional distillation, products were obtained as follows:

(a) Between 87-90°: Butylamin

(b) Between 96-98° : Amylamin

(c) Under 100°: Hexylamin

(d) Between 198-200°: Hydrotoluidin

The distillation was continued to 215°. After cooling, the dark brownish residue was extracted with ether. The ethereal extract was evaporated and the residue dissolved in dil. hydrochloric acid sol. To this sol., platinic chlorid was added. The resulting precipitate was the chloro-platinate of aselin, an alkaloid of the composition indicated by the formula C25 H32N4, present in the original oil in small quantity. From the mother-liquor the chloro-platinate of morrhuin was obtained, to which the formula C19H27N3 was ascribed. It is possible that these two substances were secondary products of the distillation.

A second paper dealing with this subject was published by Iscovesco (2), who claims to have isolated a lipoid from cod liver that possesses all the known therapeutic properties of cod-liver oil.

The writer has been working on this subject during the past year in the hope of isolating vitamine-like substances that might account for the action of the oil in curing rickets and accelerating growth. The results of the preliminary work are given below.

Regarding the action of cod-liver oil there are two distinct views: that of the writer (3), who attributes the action to the presence in the oil of a vitamine-like substance; and of Osborne and Mendel (4) who regard the action as due to the special nature of the fats in the oil. The work of Gautier and Mourgues, and that of the writer, show that cod-liver oil contains a certain amount of organic bases, a fact which must be taken into account.

If the writer's view is correct, it might be advisable to administer, in rickets, cod-liver oil that is less purified than that used at present. Also, it should be possible to administer an extract of the organic bases without the oil. Such products have been obtained and will be tested on animals at the earliest opportunity. As experimental animals, chickens will be used, which develop in captivity, on a uniform diet, a condition resembling rickets. Some of the results obtained with cod-liver oil have already been published elsewhere (3).

The cod-liver oil subjected to fractionation was a very dark crude oil. A second sample of crude oil, though lighter in color, gave much less extractive material than the darker one. In the first case the oil was extracted by a method very similar to that