CHAPTER VI

THE CHEMICAL AND PHYSIOLOGICAL PROPERTIES OF THE VITAMINE

While the chemists have not yet been able to isolate and identify the various vitamines they have succeeded in demonstrating many of the properties of these substances and it is the knowledge of these properties that has enabled us to produce concentrates and conduct tests. Another practical consideration involved in this matter of properties lies in the effect of cooking and commercial methods of food preparation, for not only must we learn where the vitamine resides but how to prevent injury or destruction in our utilization of the source.

The properties of the vitamines may therefore be grouped under two heads: first chemical properties and second physiological properties.

I. CHEMICAL PROPERTIES OF VITAMINE "A"

a. This dietary factor's presence in butter fat and egg yolk fat indicates its solubility in the fat and it would naturally follow that the fat solvents would suffice to remove it with the fats when food sources are treated with such a reagent. Experience has shown however that while ether extraction applied to butter or egg yolk removes the vitamine with the fat this process fails when it is applied to vegetable sources such as cotton seed, corn germ, spinach, lettuce, etc. Neither does the cold or hot press method of oil extraction liberate the vitamine with the oil. Recent experiments by Osborne and Mendel, to which we have previously referred, have shown that preliminary treatment of vegetable sources with alcohol seems to loosen the bond between the source and the vitamine and that when this binding is once loosened subsequent ether extraction will take the vitamine out. That the binding is not difficult to break is shown by the fact that when vegetables are eaten as a source of vitamine the body is able to separate the complex. It is further evident that the body does separate this complex and stores it in animal fat from the experiments with cow feeds and feeding. Milk for example is rich or poor in vitamine according to the supply of the latter in the food given to the cow. The only logical conclusion to be drawn from this observation is that the cow does not synthesize this factor but splits it off from the food source and then, since it is fat soluble, is able to mobilize it in the butter fat of the milk or to a more limited extent in the body fat. This observation as to the dependence of milk content upon food has been confirmed in the case of nursing mothers and suggests the need of especial attention to the diet of the mother during the lactating period.

b. It has been generally assumed that the "A" vitamine is comparatively stable to heat. Sherman, MacLeod and Kramer state that "dry heating at a temperature of 100°C. with free access of air, only very slowly destroyed fat soluble vitamine." Osborne and Mendel reported that butter fat treated with steam for two hours and a half did not appear to have lost its value as a source of this vitamine. Drummond's earlier work with fish oils and whale oils seemed to confirm this conclusion. Sherman and his co-workers cited above put it this way: "The results thus far obtained emphasize the importance of taking full account of the time as well as the temperature of heating, and of the initial concentration of the vitamine in the food, as well as of the opportunity for previous storage of the vitamine by the test animal." More recent work by Steenbock and his co-workers in America shows that these earlier results are incorrect in the case of butter fat and that twelve hours exposure of butter fat to 100°C. may, under certain conditions, destroy the efficiency of that substance as a source of the vitamine. Drummond and other English workers have confirmed Steenbock in later experiments. Their work has shown that the presence or absence of oxygen is a factor, which may determine the extent of destruction of the vitamine. Heat alone is of very limited effect but when sources are heated in the presence of oxygen destruction of the A vitamine may be very rapid. Drummond attributes the absence of the A vitamine in lard to the oxidation that takes place in the commercial rendering of this product. We must conclude therefore that while the vitamine may be destroyed by continuous exposure to a temperature of 100°C. the effect is largely determined by the nature of the process and the way the vitamine is held in the source. Cooking of vegetables therefore will not as a rule result in appreciable destruction of this factor.

c. The process of hydrogenation used in hardening fats appears to completely destroy the vitamine, hence the many lard substitutes now in use must in general be considered "A" vitamine-free regardless of the content of "A" in the fats from which they are derived unless they have been made by blending instead of hydrogenation.

d. Acids and alkalies have apparently little effect on this particular vitamine.

It may be well to state here however that owing to variability in behavior with variation in conditions it is dangerous to draw too general conclusions and until a given source has actually been investigated under specific cooking conditions one should not rely too strongly on analogies based on comparative experiments. This statement applies to all vitamines and presents one of the live subjects of investigation for the cooking schools and the food factories.

e. Little has been learned further about the chemistry of this substance. [Footnote: Since the above was put in type Steenbock has shown that the A vitamine resists saponification and that by saponifying fats which contain the A it may be possible to secure a fraction rich in the vitamine and free of fat.] Butter fat, nitrogen free and phosphorus free is shown to carry the vitamine and it is therefore assumed that the vitamine lacks these elements. It has been claimed that it may be removed from butter fat by prolonged extraction with water but this has not been confirmed by more recent experimenters. Steenbock was the first to call attention to the association of the A vitamine with yellow pigment in plant and animal sources. Butter, egg yolk, carrots, yellow corn contain it while white corn and white roots are less rich in this vitamine. This observation suggested the chemical relation between the vitamine and carotin. It has however been shown by Palmer and others that carotin is not vitamine A. This association of the pigment with the vitamine is therefore apparently a coincidence and this clue has failed as yet to throw light on the chemical nature of vitamine A.

II. THE CHEMICAL PROPERTIES OF VITAMINE "B"

When Funk first studied this substance he conducted all his evaporations in vacuo from fear that higher temperatures would prove destructive. Subsequent investigation however has shown that 100° has very little if any destructive effect if the vitamine is held in acid or neutral solution. Temperatures between 100° and 120° maintained in an autoclave at 15 pounds above normal pressure do tend to slowly destroy the factor. The extent of this destruction also varies with the character of the crude extract. In general, then, there is little fear of injuring this vitamine in ordinary cooking temperatures if the use of alkali is avoided.

The effect of alkali depends upon the temperature to a very marked degree. Osborne has recently reinvestigated this matter and finds that in the presence of a 0.1N solution of alkali at 20°C. there is very little destruction but that raising the temperature to 90°C. brings about a marked destruction. Seidell has shown that if the vitamine is absorbed by Lloyd's reagent and this reagent be then extracted with dilute alkali the vitamine passes into the alkaline solution. If the latter is neutralized quickly it is possible to recover most of the vitamine by this method. The effect of alkali becomes of practical importance to the housewife because of certain cooking habits. I refer to the well known practice of adding soda to the water in which vegetables are cooked to soften the vegetable and accelerate the cooking. Daniels and Loughlin in this country investigated this matter and came to the conclusion that this procedure did not produce enough destruction to be dangerous. Later the matter was studied by Chick and Hume in England and these investigators brought out a feature that had perhaps been overlooked in the previous work. Their point was that in ordinary feeding tests the results merely tell whether there is enough vitamine present to produce normal growth. Hence if the substance tested has much vitamine, a large part of it might be destroyed and this fact not appear in the test because enough might still be left to induce normal growth. By reducing the amount tested so that it was just adequate for normal growth and then applying the soda-cooking experimentation they showed that this method of cookery does do serious harm to the vitamine. From the practical point of view it is of course sufficient to show that enough is left after a cooking process to suffice for normal growth when the substance is taken in the portion sizes ordinarily eaten. The effect of alkali deserves more attention on the part of cooks and food preparateurs and we need more data concerning the minimal dose necessary to protect the human animal.

In neutral and acid solution it is perfectly safe to assume little destruction of this vitamin through heat and it is now common practice to boil sources with the extracting reagent and to use the steam bath freely to concentrate and evaporate these extracts. We have recently investigated the effect upon cabbage of cooking in a pressure cooker at eight pounds pressure. The cabbage so cooked, when dried and mixed so as to form 10 per cent of a basal vitamine free diet, yielded all the "B" vitamine necessary to produce normal growth in rats.

The very name of this vitamine indicates its ready solubility in water. It is also soluble in 95 per cent alcohol and either of these extractants may be used to obtain the vitamine. It is not readily soluble in absolute alcohol and 95 per cent is not as good an extractant as water. Substances rich in the vitamine apparently yield the latter more readily if they have first been subjected to autolysis or if the extracting fluid is acidified. Funk was the first to show that yeast produced a greater yield if it was allowed to autolyse before extraction with alcohol. However, Osborne and Wakeman have produced a method of treating fresh yeast by boiling it with slightly acidified water which seem as efficient as autolysis in the yield produced.

The various methods of extraction now in vogue have already been discussed in Chapter II and need not be repeated here. In general it is apparent that to obtain concentrates of high potency it is permissible to employ temperatures of 100°C. if we will maintain an acid or neutral reaction but that alkali should be avoided wherever possible and when its use is imperative the temperature must be kept below 20°C. or destruction will result. In applying this rule to cooking operations the results should be determined by direct tests rather than by assumptions based on these generalizations. It should also be noted that the alkalinity of a solution should be determined on the basis of hydrogen ion concentration and not on amount of alkali added since many substances have a marked buffer reaction.

The water-soluble "B" is not only soluble in water but can be dissolved in other reagents. Thus McCollum has shown that while benzene is of little value as an extractant of this vitamine, if we will first extract the vitamine with alcohol or water and deposit this on dextrin by evaporation it is then possible by shaking the activated dextrin with benzene to cause the vitamine to pass into solution in benzene. Voegtlin and Meyers have recently shown that it is soluble in olive oil and in oleic acid and their data suggest a new means of concentrating the substance which may be of value in tracing its character.

The "B" vitamine is relatively easily absorbed by finely divided precipitates. We have already referred to the use of fuller's earth for this purpose by Seidell. This adsorptive power sometimes manifests itself in the treatment of plant extracts. A watery extract of alfalfa can be made to throw down its protein complex by diluting it to 40 per cent with alcohol. Osborne reports however that this process frequently removes the vitamine also which appears to be thrown down with the precipitated material. This adsorptive power therefore often appears as a difficulty in the handling of the substance as well as a means of extraction. We have used Osborne's method with alfalfa extracts and find the above result is not by any means invariable, for in some of our extracts we retained the greater part of the vitamine. Kaolin and ordinary charcoal are not very good adsorbents but the latter can be activated to serve this purpose.

The elementary nature of the "B" vitamine remains a mystery. Extracts which contain it show the presence of nitrogen. Funk's earlier researches on yeast and rice polishings both yielded crystalline complexes which he analysed. His data on this subject follow:

A. The yeast complex

Crystals melting at 233°C. consisting of:

I. A complex melting at 229°C. and forming needles and prisms nearly insoluble in water and with the apparent formula of C_24H_19O_2N_5.

II. A complex melting at 222°C. and soluble in water. Formula
C_29H_23O_2N_5.

III. Nicotinic acid melting at 235°C. C_6H_5O_2N.

B. The rice complex

Crystals melting at 233°C. consisting of:

I. A complex melting at 233°C. and with a formula of C_26H_20O_9N_4.

II. Nicotinic acid melting at 235°C. C_6H_5O_2N.

Funk held at the time that the possible nature of the compound was:

HN
| \
OC C_16H_18O_6
| /
HN

It was this idea that led him to call it an "amine."

We are unable at present to report any nearer approach to the elementary analysis and all attempts at purification have shown a tendency to make the active substance either disappear entirely or else distribute itself over the several fractions instead of concentrating itself in one. Its basic nature seems to be well established by its behavior with phosphotungstic acid and its ready adsorption by carbons activated to take up basic substances.

III. THE CHEMICAL PROPERTIES OF WATER-SOLUBLE "C"

The properties of this newest member of the family are still less defined. All are agreed that it is much more sensitive to heat and alkali than the other two. Temperatures above 50°C. are usually destructive though the time factor is extremely important as well as the reaction. Hess for example has found that the temperature used to pasteurize milk continued for some time, is more destructive to the vitamine than boiling water temperature continued for only a few minutes. The extent to which orange juice and tomato juice will resist high temperatures indicates the protective action of acids to be considerable.

Dr. Delf's experiments at the Lister Institute were especially directed to the behavior of this vitamine in cabbage. She first determined the minimum close of raw cabbage required to prevent scurvy in guinea pigs and found that it was less than 1.5 grams and more than 0.5 gram daily. When the cabbage was heated in water at 60°C. for an hour, symptoms of severe scurvy were just prevented by 5 grams of the cooked cabbage fed daily. By heating at 70°, 80°, 90° and 100° for the same length of time the 5 grams of cooked material could be made non-effective as a preventive. Her conclusions are that when cabbage is cooked for one hour at temperatures ranging from 80° to 100°C. the cabbage leaves lose about 90 per cent of the antiscorbutic power originally held by the raw equivalent. Sixty minutes at 60° or twenty minutes at 90° to 100° resulted in about 80 per cent destruction. Dr. Delf calls attention also to the fact that the effect of the heat is increased to only a slight degree by rise in temperature. Assuming that the effect of the rise is orderly, a temperature coefficient of 1.3 is indicated for each rise of 10°C. This low result suggests to Delf a contradiction to any theory which imputes to the vitamine enzyme or protein-like qualities and on the other hand suggests that the substance is much simpler in constitution. Her results also confirm Hoist and Fröhlich as showing its great sensitiveness at temperatures of 100° and below and obviously have a direct bearing upon cookery methods.

The substance is soluble in water and passes through a parchment membrane or a porcelain filter. Unlike the "B" it is apparently not adsorbed by fine precipitates such as fullers' earth or colloidal iron. Harden and Zilva showed that when a mixture of equal volumes of autolysed yeast and orange juice is treated with fuller's earth the "B" is removed and the "C" left unaltered. Eddy and La Mer have treated orange juice with fullers' earth and then tested the filtered off juice as cure and preventive of scurvy in guinea pigs. Their results showed that 6-2/3 cc. of the treated juice was curative, hence the loss due to adsorption must be less than 60 per cent to 70 per cent. Harden and Zilva were among the first to state that the vitamine is much more stable in acid than in alkali. They have shown, that even 1/50 N sodium hydrate at room temperature has a rapidly destructive effect. On the other hand Delf showed that when 0.5 gm. citric acid is added to the water in which germinated lentils are boiled, the loss of the antiscorbutic properties is, if anything, greater than when no addition of acid is made. She therefore concluded that in cooking vegetables there should be no addition of either acid or alkali to the cooking water if one wishes to conserve this vitamine. Sherman, La Mer, and Campbell have been engaged in experiments bearing on this point throughout the past two years. Some of their results have recently been published and their observations are worthy of special attention from their bearing on the character of reaction of the vitamine in general. They first proceeded to determine the amount of filtered tomato juice just necessary to produce scurvy in degrees extending from no protection to complete protection and they also constructed a basal diet which is apparently optimum in nutrients and all other factors except the "C" vitamine. They found that at the natural acidity of tomato juice (pH 4.2) boiling for one hour destroyed practically 50 per cent of the antiscorbutic power and by boiling for four hours they destroyed 70 per cent, which indicates that the curve of the destructive process tends to flatten more than that of a unimolecular reaction. This result was confirmed by heating experiments conducted at 60°, 80° and 100°. In all cases the temperature coefficients are low. (Q_10 equals 1.1-1.3) confirming Delf's results. When the natural acidity of the juice was first neutralized in whole or in part, the juice then boiled for an hour and immediately cooled and reacidified, it was found that at less than half neutralization (pH 5.1-4.9) the destructive effect of an hour's boiling was increased to 58 per cent. When alkali was added to an initial pH 11 (about N/40 titratable alkali to phenolphthalein) which fell to 9 during the hour's boiling the destructive effect was about 65 per cent. When reacidification was omitted and the neutralized boiled juice stored in a refrigerator for five days before using the destruction increased 90 to 95 per cent. These particular observations seem to confirm the view of Harden and Zilva that the vitamine is especially sensitive to alkali. Hess has recently reported that oxygen is destructive to this vitamine.

IV. PHYSIOLOGICAL PROPERTIES OF THE "A" VITAMINE

Most authorities are now agreed that both the "A" and "B" types are essential to growth. Rohmann still holds out against the vitamine hypothesis. McCollum has recently pointed out that while rats do not have scurvy it does not at all follow that the absence of the "C" in their diet is immaterial, but that the contrary is true. Failure to grow, then, may manifest itself as a result of the absence of either of the first two types and possibly is affected by the absence of the "C." We have already seen how this failure may be utilized to measure the vitamine content of a source. The absence of the "A" type however may also manifest itself in another way, viz., by the development of an eye disease which McCollum first designated as xerophthalmia or dry eye and which the British authorities prefer to designate as keratomalacia. The failure of this result to always follow the absence of the "A" type in the diet has led some to question the specificity of this disease. While the infection of the eye is due to other agents the sum of the evidence supports McCollum and points to the absence of "A" as the true predisposing cause of the disease. Bulley, basing her claims on a study of some 500 rats fed on a synthetic diet, claims that the eye condition is not primarily due to a dietary deficiency but to an infection resulting from poor hygienic conditions. In reply to her contentions Emmett has reviewed his own data and presents them in the following summation:

_________________________________________________________________________
| | | |
RAT | KIND OF VITAMINE | NUMBER CASES | POSITIVE CASES | PER CENT
GROUPS | ABSENT IN THE RATION | REPORTED | OF XEROPH- | POSITIVE
| | | THALMIA |
_______|______________________|______________|________________|__________
| | | |
A | Fat-soluble "A" | 122 | 120 | 98
B | Water-soluble "B" | 103 | 0 | 0
C | None | 216 | 0 | 0
_______|______________________|______________|________________|__________

In these groups special hygienic measures were taken against infection. Furthermore repeated attempts were made to transmit the eye disease by using sterile threads, passing them carefully over the edges of the sore lids and then carefully inoculating the eyes of other rats. These attempts resulted negatively in all cases where the inoculated rats had plenty of the "A" vitamine. Treatment of advanced cases of sore eyes with a saturated solution of boric acid and also with a silver protein solution failed to relieve the condition while as little as 2 per cent of an extract containing the "A" vitamine when added to the ration, speedily resulted in cure and increase of weight. These results combined with similar data compiled by Osborne and Mendel seem to refute Bulley's contentions and to justify our acceptance of xeropthalmia as a specific vitamine deficiency disease.

Osborne and Mendel data

Total No. No. with eye symptoms

Rats on diets deficient in A vitamine . . . . . . . . 136 69 " on diets " " B " . . . . . . . . 225 0 " on diets otherwise deficient . . . . . . . . . 90 0 " on " experimental but probably adequate . 201 0 " on mixed food . . . . . . . . . . . . . . . . 348 0 ____ __

Totals . . . . . . . . . . . . . . . . . . . . . 1000 69

On the other hand all workers know that rats often do develop and grow well for a considerable period of time on a diet free from the "A" and without manifesting the eye disease. The British authorities explain this by assuming that animals have the power to lay down a reserve of this vitamine on which they can draw in emergency. Sherman and his coworkers confirm this power to store the vitamine. Others have been led to explain their results as due to contamination of the basal diet. Daniels and Loughlin recently maintained that the commercial lard used in basal diets and assumed to be "A" vitamine-free was supplied with sufficient of the "A" to produce growth and prevent eye disease. Their views have failed of confirmation by Osborne and Mendel. It is evident therefore that these occasional lapses from specific response to absence of the "A" vitamine need further elucidation. It is equally manifest that in the majority of cases the absence of the "A" will result in both stunted growth and xeropthalmia. The appearance of the eye disease may be taken however, as a sure indication of the absence or deficiency in the "A" vitamine.

V. PHYSIOLOGICAL PROPERTIES OF THE "B" VITAMINE

Beri-beri is a disease that is described clinically as a form of severe peripheral neuritis and may appear in two well marked forms. In one type there is great wasting, anesthesia of the skin and finally paralysis of the limbs. In the other, the most marked symptom is excessive edema which may affect trunk, limbs and extremities. In severe cases the heart is usually involved and death may occur suddenly from heart failure.

Most observers assume that the antineuritic vitamine discovered by Funk and the water-soluble "B" are identical. This view is based on the fact that when sources which yield the water-soluble "B" in rat feeding are tested for antineuritic power these sources are apparently parallel in antineuritic power and growth production. Furthermore rats deprived of the water-soluble "B" develop polyneuroses identical in symptoms with those shown by rats and pigeons when the latter are placed on a polished rice diet. The British Medical Board has compiled the following table to support this view:

Table compiled from pages 35 and 86, British Medical Research Committee Report

Emmett has recently opposed this view and suggests that while the antineuritic factor and the growth factor are found in the same sources and have much in common it does not follow that they are identical and that his experiments tend to show that there are marked differences which suggest that the "B" type is not a single entity but a group. Mitchell has summarized very well the controversial phases of this question with an impartial review of the facts. One of strongest of the opposition arguments lies in the failure of milk to cure beri-beri except when administered in large quantities. This objection has been partly allayed by data bearing on the relation of the milk content to the food of the cow. Hess, Dutcher, Hart and Steenbock and others have adduced sufficient evidence to show that the vitamine content of the milk of a cow is largely determined by the cow's food and as a consequence the milk may be very poor in vitamine. It is obvious then that the failure of the milk to cure beri-beri in a given case might be due to this cause and not to lack of identity of the curative with the growth factor. Osborne and Mendel have also shown that milk in general must not be classed among the rich sources of the vitamine, even when the cow's food is rich in vitamine. The principal facts in the controversy have been presented and at present the evidence for regarding the vitamines identical seems to be preponderant.

Recently Auguste Lumiere in Paris has put forth the view that polyneuritis is not merely a vitamine deficiency disease but a nutriment deficiency disease. He reports that he fed birds on a starvation diet, but with plenty of vitamine "B". These birds developed polyneuritis and were cured by adding to the diet plenty of polished rice. The view he wishes us to take is that all factors must be present and that the absence of the nutriment is as important as the absence of the vitamine.

In the field of nutrition the absence of the "B" type is particularly marked by the behavior of the deprived animal. Rats transferred from a vitamine-free diet to one containing the "B" only, make a much more rapid recovery toward normal (even in the absence of the "A") than do animals transferred from the vitamine-free diet to one containing the "A" and not the "B". This initial jump from addition of the "B" will not continue long in the absence of the "A", as a general rule. Hess believes that in some of his infants he was able to show markedly successful growth on the diet deficient in the "A" but rich in the "B". It is not certain however that his diets were sufficiently devoid of the "A" factor to be declared "A" vitamine-free and we know little of the amount of the "A" necessary to normal infant growth. All results however show that both "A" and "B" are necessary to growth production and though the term growth vitamine was applied to the "A" originally the distinction is one that should be rejected, for both "A" and "B" and possibly "C" are all entitled to this name.

The manner in which the "B" vitamine acts is still obscure. Voegtlin some time ago tried to demonstrate that it was identical with secretin and stimulated pancreatic flow. Recent work at the Johns Hopkins University by Cowgill and by Aurep and Drummond in England has failed to confirm this. One of its most marked immediate effects is increase in appetite. Karr in Mendel's laboratory has shown that dogs which refused their basal diet would resume eating it if they were allowed to ingest separately a little dried yeast. Karr studied the metabolism of these dogs as regards nitrogen partition but the results give little data that is explicatory of the behavior of the vitamine. In 1915 the author was able to bring about marked immediate improvement and the ultimate recovery of a number of infants who were of the marasmic type by merely increasing the "B" vitamine content of their food. In these cases the vitamine was carried by Lloyd's reagent and administered mixed with cereal, or the crude extract was combined with the milk. The pancreas of the sheep was the source used. In these cases the growth curve changed abruptly from a decline to a sharp rise and this increase in weight continued and was accompanied by all the other signs of improved nutrition including increase in appetite. The change in the growth curve from decline to rise was accomplished without increasing or changing the basal diet but as the appetite increased the food had naturally to be increased to keep pace. In these cases the effect of the vitamine was to enable the child to utilize its normal food and to increase its appetite for it. This action certainly suggests stimulation of digestive glands. It also showed that even though the diet may contain the vitamine as was the case in the milk fed to these children the addition of the vitamine in concentrated form often gives an upward push that the food mixture fails to accomplish. Daniels and Byfield have recently confirmed the effect of increased "B" in infant growth. Cramer has suggested in a paper published recently in The American Journal of Physiology that the fatty tissue about the suprarenals may be a depository of vitamine and that in the absence of vitamine this tissue loses its supply and that this is the explanation of lessened activity of that gland in certain metabolic disturbances. This idea tends to support the idea that vitamines are gland stimulants or hormones and the word food hormone has been suggested to describe them on that account. A few years ago Calkins and Eddy tried to determine the effect of the vitamine on the single cell by use of the paramecium but the results of the experiments failed to show a vitamine requirement on the part of these animals. McDougall has recently suggested that the vitamines produce their effect on yeast cells by increasing hydration. Unfortunately nearly all stimuli which produce growth are accompanied by hydration effects and it is difficult to feel that this is a specific vitamine effect although without denying the possibility. Dutcher has tried to show that vitamines have a relation to oxidation effects. He observed that the issues of polyneuritic birds showed a marked reduction in catalase and that this catalase was restorable by curing the birds with vitamine. The main difficulty lies in the conflexity of factors that function between cause and effect.

[Illustration: FIG. 8. THE EFFECT OF VITAMINE B ON A MARASMIC INFANT

1. On the twentieth day the patient developed a cough. 2. On the twenty-first day the cereal was reduced from three times a day to twice a day. The patient cried during the night. 3. On the twenty- second day the stools showed free starch. 4. On the twenty-third day an anal abscess was opened. The stools continued to show free starch until the twenty-fifth day. 5. On the twenty-fifth day the stools showed soluble starch but no free starch. 6. On the twenty-seventh day the appetite was good and there was no starch. 7. From the twenty-eighth to the forty-third day no starch was observed in the stools. 8. On the thirty-first day the patient developed a cough. 9. From the forty-ninth day to the time of discharge three tablespoonsful of orange juice were given daily. 10. On the seventy-third day the patient developed a bronchitis and mustard paste was applied every four hours up to the eighty-fourth day.

V1 = From the twenty-first day to the forty-third day the patient received each day 2 grams of Lloyd powder, activated with pancreatic vitamin. The powder was administered by mixing 1 gram. with each cereal feeding. The result was 20 ounces gain in twenty-two days, a normal growth.

V2 = After a period of ten days without vitamin, during which the patient settled down to a level growth curve, the treatment described under V1 was resumed. This was continued from the fifty-third to the seventy-sixth day. The result was the resumption of growth but at a slower rate; 8 ounces were gained in twenty-three days. During the latter part of the period the patient developed a bronchitis. At the end of this period the patient was placed on a whole milk formula. From that time to the time of discharge the patient grew normally.—From the American Journal of Diseases of Children, 1917, xiv, 189.]

[Illustration: Effects of Vitamines on Growth FIG. 9]

These views are at best speculations. The literature is singularly lacking in detailed metabolic analyses of excreta of animals during vitamine stimulation and we know nothing of the possibilities of overdosage, for in all the work done it has been generally assumed that the presence of an amount greater than that necessary to produce normal growth is not material.

The exact manner of the vitamine's action then remains to be determined and it is obvious that this solution will come much more rapidly if we can first identify the substance chemically.

VI. THE PHYSIOLOGICAL PROPERTIES OF THE "C" VITAMINE

The steps that led to the acceptance of scurvy as a vitamine deficiency disease have already been discussed and show how the vitamine acts in such a disease. Practically all the work done with this vitamine to date has been concerned either with dosage or with reaction to heat, drying, etc. The only paper that we have seen that suggests another function than antiscorbutic power for this vitamine is the one by McCollum and Parsons in which they suggest that even in animals where scurvy does not exist, the presence of this factor may be necessary to normal metabolism. The following table gives some of the data compiled by the British workers as to the antiscorbutic power of various sources:

Table compiled from, page 44, British Medical Research Committee Report

A glance at this table shows the richest sources (see also table on page 59.) To these must be added canned tomato juice which Hess has shown practically equal to orange juice in efficiency and uses with infants in the same quantity. This discovery is of great value in instances where the cost of orange juice is often prohibitive.

La Mer and Campbell have presented some evidence to show that the antiscorbutic vitamine has a direct effect upon the adrenal glands. In their scurvy cases they find definite evidence of the enlargement or hypertrophy of this organ. Whether it affects other organs or not it remains to be shown.