Status of human requirements for vitamin; MK, Horwitt,? PhD. ABSTRACT The development of adult range of the requirement for the tocopherols which depends so much on past and pres dietary habits makes it dificult to-fix on a single figure for use in composite tables hutzitional requirements. Attemps to relate the requirement to the amount of polyansstura acids (PUFA) in the diet only has many disadvantages. To allow for synthesis and selec storage of PUFA in tissue lipids even when the dict is very low in such lipids, a baseline Fig for tocopherol needs is suggested to which the additional requitements for PUFA inthe diet ‘added in order to obtain the requirement. Evaluation of long-term accumulations oF tino acid in depot fat of adult men on different diets indicates that one should probably consi the percentage of PUFA in the dietary fat separately from the total amount of PU consumed. Accordingly, i Is suggested that in calculating the requirement for vitamin E, eq ‘weight be given to estimations ofthe percentage of PUFA in the dietary fats and to the amo) ‘of PUFA in the diet. The result of such a calculation Is then added tothe bast figure 0 obt the total dally requzement. The formula used to calculate the tocopherol equivalent requi per day is 0.25(% PUFA + g PUFA) + 4 = da-tocopherol equivalent (mg). Nutr, 27: 1182-1193, 1974, ‘The purpose of this report isto evaluate and to try to update the daily minimum require- ‘ments for vitamin E that may be determined from the available experimental evidence. There will be little discussion with respect to whether ‘or not larger than minimum daily ingestions of the tocopherols can have a beneficial protective fr physiological effect as we would rather this remained an open subject. - ‘The minimum human requirement for the tocopherols may not be much different from that for experimental animals after corrections are made for metabolic size (1). The apparent differences may be related to the fact that such animals gre usually in an early stage of rapid growth. Giis generally agreed that it is more ifficult to produce a vitamin E deficiency in an adult animal than in one tha is very youn ‘This may be due to the stores of lipids an tocopherol that gradually accumulate as the Janimal matures The adult rabbit can be used for deficiency studies that involve the muscles but it is more difficult to create. similar deficiencies starting with adult rats. This is related to the higher content of linolenic avidin adult rabbit tissue lipids dre to the higher dietary content of alfalfa. meal lipids and soybean flour lipids which may contain over 40 nis The American Journal of Clinical Nutrition 27: OCTOBL® 1974, pp. BE Am. 1.0 ‘and 12% linofenates, respectively. The lipids ‘experimental rat diets usually ar¢ less unsatur ted in terms of potential peroxidizability ev when the linoleic acid content is relatively hig Lard and corn oll so often used in experiment is deficient in vitamin E, which have-abo 12 and S0% linoleic acid, respectively, conta very little linolenic acid, The fatty acid comp: sition of animal fats is, of course, depende fon what the animal was fed. Thus, lard linole acid may vary from 5 to 25%. Furthermore, th attention being paid to unsaturated lipic should not imply that these are the onl compounds protected by the tocopherol Other compounds susceptible to deterioratio by chain reactions that are inhibited by th tocopherols could just as well be included this discussion. In Orgamrinesraic nm man. the possibitir that irreversible damage may Le produce makes it unthinkable 2» ws for human 20 which provide, levels ui tocopherol for prolonged pi time. Inthe Elgin project on vitamin | From the Department of Biochemistry, St. Lo University School of Medicine, St. Louis, Miso 6108. sn BinchemistryHUMAN REQUIREMENTS FOR VITAMIN E jaromonts (2=4},-we were cautious not to ld a completely deficient diet despite the sonable certainty that it would take at least ats to deplete the reserves in the average viously well-fed edult. It is extremely doubt- tiatit will be possible to repeat such a term nutritional deticion.y study in the seeable future. One point should be cari | for those who do not have experience in fies which evaluate requirements of nut nal components of the diet. One would-tearn y little more about requiremes:is than to firm that a nutrient is-required, if all of the ary component in“question was taken out the diet. Ratker, the objective should be to »-detéimiine the minimum level at which, er prescribed circumstances, pathology is fented. The closer you are below this imum requirement in the experimental diet, longer it takes to achieve the information red. less satisfactory procedure that entails far ter risks is to create a deficiency ina ter time and then add increments of the ponent being studied to the diet to the t where deficiency symptoms are reversed adequacy is interpreted. One of the 2ulties of this proceduse is that the reversal ‘™mptoms, which should be repaired as soon vssible, may require a larger dose than one 5 to estimate true minimum requirements. +s long supplementation periods with mini- doses are planned one may interpret the rement on the higher sid, the study of vitamin E requirements, the em is particularly difficult because of the nrminate floating variables, ie., the nt of petoxidizable unsaturated” com- 4s in the. tissues, the oxidizable compo- in the diet and the time requited to use reserves of the tocopherols which, in depends on the level of peroxidizable wales in the tissues. Thus, one can ze an extreme situation where 5 mg of pherol a dey could supply the minimum. rment for a period of years in a previ- well-fed adult with low tissue levels of tates, whereas at other levels of tissue let composition 15 mg would not be 1. And then there is the problem of the © need for a safety factor for contingen- vor example, few, if any, individuals show any recognizable signs of scurvy or 1na3 other related pathology if their det was limited to 10mg of ascorbic acid (5) @ day. But certainly, even in this far tess complicated situation one could hardly recommend so lite vitamin Cas @ requirement, [in the Sheffield experiment (6), cures of vital A deicleney as determined by evaluating dark adaptation were obtained with a level of 1,300 10 of vitamin A and only 3 out of 16 men showed Sigs of deficiency after more than» yeat ona et which had’ only’ traces of Wlamin, A activity. But, knowledgeable workers in this field would hesitate to. lower the. curtent requirement to les than 2.500 IU of vitamin A per day In the first Elgin Broject (7) a group of zen who received only 04 mg of thiamin and O85 mg of rbofavin a day showed lite, if any, signs of deficiency and ina later study on niscindryptophan (8) only 9 niacinequivalents produced na physiological sins of pathology. Revertheless, thas been the judgment of the best minds in nutrition to increase the sg: gested requirement for thiamin ribolevin, and nlacin above these amounts. 7 One: of the major obstacles in rationalizing some of the experimental data on vitamin E wi petal etry orbs at relatively dificult to incorporate much Sore than 20 IU of a-tocopherol into some diets using normal foods that are compatible with cour eultural patterns. I is eurent conventional wisdom that diets need not be supplemented ‘ith vitamins ifthe components are properly chosen. We all know of exceptions to. thi dogma which made it logial to add some favaoluble vitamins to milk, some. of the watersoluble vitamins to grain products and of iton to bakery products but by and lage we like to beleve that we need not supplement propery selected diets with vitamins to achieve consumption of minimum requirements. The rationalization or food fortifications 1s that they are replacements for amounts removed by manufacturing processes Since the soutce Of dtectocopherol fs fom food and feed products being processed for human and animal con- sumption, vitamin E losin food preperation may not be much different in ths respect Most vitamins other than vitamin E have been shown. t0 be intimately felted (0 ene 2ymatic or hormonal effects or tobe in some ‘manner a significant part of a chemical equa. tion. The tocopherclt may also some day bees } shown to have a similar specific function but for the time we are stil looking.) Without | belittling the possibilty that tocopherol has other functions, when one considers all the physiological systems that have peroxidizable lipids in the presence of traces of oxygen and metals which could catalyze peroxidations, if tocopherol had not evolved, nature would have had fo invent a different antioxidant to do the Job One of the, characteris of feexadie traps that can delay the initiation and possibly the rate of reactions that may be undesirable, is that the more antioxidant that may be present in a given mixture, the greater the potential of it trapping an errant free radical. The same analysis cannot be made for vitamins with functions that can be described in stoichio- metric equations where more than a preseribed amount has no additional effect. Therefore, some of us are led to wonder, in the absence of any hard proof, whether.’ more than the established minimum requirement of this anti- oxidant ean have 2 biological consequence. About all we as scientists can do while waiting for a final answer is to encourage more work and keep an open mind. On the other hand, claims for physiological effects which are not supported by controlled experiments should not be encouraged. One of the clasical methods of evaluating the need for a given nutritional substance depends on the relationship between the amounts consumed by population groups and their relative states of health. For example, if xerophthalmia becomes prevalent in a given ‘geographic area one would try to estimate what amounts of vitamin A were being consumed by the population group involved and relate such information to that from surveys of populs- tions which do not show manifestations of xerophthalmia. Such data are useful but they five only broad ranges of requirements. Addi- tionally, apparent optimum health based upon the absence of physiological dysfunction using old standards are not always reliable. For example, consider how some of the Asiatic populations increased the rate of their growth when they improved the balance of amino acids in their diet. Without drawing any final conclusion whether the increase in height of the present generation of Japanese is good or bad, it isa fact that there was little reason to suspect that the Japanese were in less than optimum HORWITT health before they started to increase the consumption of animal and fish proteins. Fc many years, the old diet was considere adequate. Population surveys of vitamin E_consumy tion have given us a broad. rango-of data... lepresentative diet recommended by the Ni tional Research Council in 1945 when analyze (9) provided only 7.59 mg of total tocopherc land 5.74 mg of actocopherol/day in 1,560 kca [This diet was relatively low in unsaturated fat ff this diet composition was extrapolated 1 ,000 Keal, the total tocopherol content ws 14.60 mg which is not much different than th 14.95 mg reported by Harris and Embree (10 to be present in American diets in 1960. Repetitive daily consumptions of tocopt cerols other than actocopherol can apparently protective to man and animals. This appears t be especially true if some reasonable portion. the total tocopherols is a-tocopherol. Tests potency which depend on rate of disappearan of different tocopherols from the tissues an Jbody Muids have shown thatja-tocopheral wil ersist in the erythrocytes Fonger than “7 ‘copherol (11) but as Bieri (12) has note frtocopherol is stored in the tissues for a long Iperiod than previously considered. Perhaps @ [greatest difficulty in evaluating the vitamin ctivity of diets is in estimating how muc factivty to allow for the other tocopherols an the tocotrienols. It is possible that these ha synergistic effects on actocopherol! Rece ‘estimations of vitamin E requirement based the d-actocopherol conten: of national copherol was ingested in the hope of obtain- 1g more complete equilibrium of a-tocopherol stribution between the blood and. the other ssues. No doubt, the analyses would have ven data that showed higher plasma tocoph- ol levels if we had waited less than 44 hours take the blood samples but previous experi ce had shown that 24 hours was not fficient time for the blood to achieve an vilibrated state, The data in Table 1 show, ite definitély, that 15 TU was not sufficient raise the plasma tocopherol levels to what is rerally considered acceptable even after 138 1s of supplementation. It should be recalled 1 at this point in time, the linoleic acid level the depot fat fatty-acids was approximately & Table 2 shows the effects of supplementing » subjects, that had been on the basal diet ‘54 months, with either 15 mg of d-ato- herol acetate or 20 mg of di-a-tocopherol ate per day. Including the amount in the + the total provided was 25 TU/day. Note the subject labeled 2a started out with the est level of plasma tocopherol in the eted group of subjects. It was not until time later that we realized that his higher ‘na tocopherol was a function of the higher of lipoproteins in his blood as indicated by nie? his higher level of plasma cholesterol (15). The slow decrease in the peroxide hemolysis test after supplementation in these two subjects might be ascribed to the high level of polyun: saturated fatty acids in the erythrocyte lipids which: at this time contained. approximately 14.5% linoleic acid which represents about an 80% increase from levels at the start of the study (3). His erythrocyte lipids aso contained 14.0% arachidonic-acid but that amount is not ‘unusual, ‘Supplementation with 30 mg of d-a-tocoph- rol acetate was not included in this series as we already had data on this level of supplementa. tion in the control group, Table 3 shows that 85 TU of vitamin E was mote than adequate as there was an almost immediate repair of both the plasma tocopherol levels and the peroxide hemolysis test results, The question as to how much pathology is produced in adult man if he is depleted-of vitamin E is difficult to answer. Unlike our previous studies with thiamin, riboflavin and niacin-tryptophan, where there was much prior experience about clinical deficiencies, knowl- edge about recovery from a deficiency of vitamin E in man was not at hand. Accordingly, supplementation was inaugurated soon after we noted the first decreases in erythrocyte life- time. There was never any doubt in my mind about the validity of these decreases in erythro- ‘TABLE 2 Effect of providing depleted subjects th 25 10 Period of Plasma supple tocoph- Peroxide Tocopherol ment, el, hemo- Subj. perday days mg/100 mi tyes, 2a 1S med-ee 0 046 75 tocopherol == «30663 acetate plus 608.40 sWindie 1306s 32 20700 30 om 10 8007s 2 20mg dhe 0 018 as tocopherol = «303879 aecatepus = § 037-70, SWindet 13 04s 22 2 oat 40 50 0s3 20 138 03735eee eS Se Te ees ei Me Cees niga NORWITT ‘TABLES Effect of providing depleed subjects with 85 10 Period of Plasma supple: focophy Peroxide ‘Tocopherot ‘ment, fol, hemon Subj. perday days) mg/l00.ml_ lysis, 3a 6Omgdo 9 030 73 tocopherol = 3.13 0 actatepus 6 0 SWindt 13 ° 2 2 50 2 138 1 30 BO mgdlo- 0 08s tocopherol «3060. ° aoetate plus, «056A Sind 13 070 2 21068 4 50 0.4 2 138089 3 cyte lifetime and although the changes were small, there was no overlapping of. data when ‘we paired depleted subjects with subjects who had adequate stores of vitamin E (16). In the years subsequent to the last Elgin project, I have thought a great deal about the human” requirement. There is apparently no difficulty in achieving the amounts recom- mended for infants but the dilemma of resolv- ing our experimental data with knowledge of what adult man normally eats inthe 20th century has given me many puzzled moments. Lour data clearly show that any emount less than 20 TU/day for adults eating moderately large amounts of corn oil should be inadequate and yet, millions of people consuming less than 15 IU ‘show no apparent need, by known criteria, for larger amounts. Is ‘the diet of convenience which has evolved in our society bordetline in tocopherol concentrations or are ‘our experimental data heavily weighted by the higher than average tissue concentrations of polyunsaturated fatty acids in the tissues?/Since it is possible that 8 mg of a-tocopherol/day could be adequate when the tissues are low in polyunsaturates and more then 20-me/day are heeded when the tissues are very high tr linoleates—as after long time consumption of safflower oil-there seems to be no single | satisfactory figure upon which to compromise. To guard man from products from which tocopherol has been removed or from its destruction in processing and storing there appears to be no alternative but to relate th [needs for vitamin Ein some manner to t amount of polyunseturates in the diet. Hov ever, in order to include fish products in an pertinent evaluation, the potential peroxidiz ability of the lipids in the diet rather than th total amount of “polyunsaturates: might -b preferred, Although we had earlier clues, w first reported that the “tocopherol requiremen appeared to be related to the amount 0 peroxidizable fatty acids ingested and/or store in the tissues” in 1960 (17). This was late ‘modified and tested in experiments by Wittin and Horwitt (4, 18) using the rate of appeat ance of nutritional muscular dystrophy as criterion but the idea has been slow i acceptance. What is needed is some simplifies tion of the previously suggested calculation: Originally, it was suggested that the rates o peroxidation of monoenoic, dienoic, trienoi fetraenoic, pentaenoic, and hexaenoic:fatt acids in terms of relative tocopherol requir ment was 0.3, 2, 3, 4, 5, and 6, respectivel ‘The separate ‘allowance for the more high! unsaturated lipids in the diet would take int consideration the fatty acids found in fish an fish products which sre becoming amo important part of some diets. Signific amounts of pentaenoic and hexaenoic fatt acids are deposited in the brain, the erythr cytes and the heart after consumption of fis lipids (19), ‘One of the more important objections whic ‘amounts of vitamin E to polyunsaturates in diet is based on the knowledge that even w the diet is very low in essential fatty acid significant amounts of polyunsaturated fat acids are found in the tissues and calculatio based on the low levels of polyunsaturates 4 the diet would not reflect what is present in th tissues.jIn chicks, even on a diet that contain only 1.2% linoleate as the only lipid, the _of the heart contained 20% linoleic acid and 9 (item acid (19).|And, it is well know that on an essentialy fat-free dict the body cd synthesize eicosotrienoic acid from oleic ac whiem>-efcaurse, can he synthesized fro carbohydrate and. protem voucces via acet (CoA and malonyl CoA) That relat2’ effects ef 'be obtained in. primates was shows ~m. tH teport by Fitch et al. (20) in which monkeys. a diet very low in fat developed vitaminHUMAN REQUIREMENTS FOR VITAMIN ¢ féeficiency along with signs of fatty acid fieficiency. The eicosotrienoic acid content of ts lipids of the erythrocytes of these monkey's as & high a5 7%, confiiing other studies by folman (21), whereas this fat 3 sentially absent in. monkeys fed Some linoleic To rllow for the syrithesis of polyunsatue ated fatty acids that do not come from dietary sources it is suggested that a minimum adult equirement_of 10 mg of d-a-tocopheiol be lowed even if the calculation of the require. rent from dietary constituents should result in lower figure. The arachidonic acid content of nimal muscle and organ fats may be higher han most nutritionists realize (22-24). At the ik of encouraging more theories regarding the enefits of vitamin E, it might be noted that eart lipids can be ‘quite unsaturated; one =port. showed concentrations of 25% linoleic cid and. 11% arachidonic acid in beef heart pids (22). Both the percentage of PUFA in the dietary t and the amount of such fat affect the degree | which tissue lipids accumulate PUFA in the sues. The depot fat lipids tend to achieve the me. percentage of linoleic acid as in the fat nsumed./ Thus when 60 mg corn oil was ded “19"the “basal diet the final mixture ntained about 35% linoleate and when this +t was fed for almost 5 years the depot fats iched a level of about 33% in the adult male ajects. Similarly, when 60 mg safflower oil s added to the basal diet to achieve about % linoleate in the dietary fats the depot fats ched approximately 50% (3, 14). Data on w much the total amount of PUFA in the t affects the rate of achieving final equi- ium are not available but it would appear t this should be as important as. the centage of PUFA jn the lipids fed. Stated ‘rently, it may take longer to reach tissue iibrium with lower rather than higher levels ‘ats in the diet, but the controlling factor in final. composition of the tissue lipids is as related to the ratios of the individual Y acids ingested as to the total amount of lipid ingested. say calculation of the vitamin E requiseii \t based only on the levels of PUFA in thd] ‘would not be complete since the tissue 1d have considerable amounts of polyun rated lipids even when the diet was practi 1189 cally deficient in DURA ll Accordie!y, some allowance must be made for cellwia: synthesis, and retention of PUFA in any Zaeulation of the tocopheral requremente The leat unsaturated diet used the lpn Project contained 60 mg of beet fat addedto the basal det. This dit provided only 2 of PUFA and approximately 3 mg of datoeceie trol After mote than 3 years on thi diet (re), ‘one subject, not included in Fig. 2, had less than 39 linoleate in his adipose nue ips. A of the subjects on this diet had. "marked increases in their erythrocyte peroxide hems, Iyss teat ests, Accordingly we hae cheeo a img of d-etocophero asthe arsoont tat should be allowed for tse synthesis of peronidnatle compounds in the male adult even when tne diets praciily deficient in PUPA, This gure of 4 has been incorporated into the following foxmula for determining the reauitement for img of a-tocopherol equivalents equed: 0.25 (PUFA +g PUFA)+ 4 me= ‘tocopherol requirement where “% PUFA” represents the percentage of | polyunsaturates in the total lipids in the diet { and “g PUFA” represents the {otal amount of | polyunsaturated fatty acids obtained by multi- plying the total dietary fat by the percentage of PUFA. The constant, 0.25, is the estimate obtained when the formula ‘is applied to the data in Table 4,!To test this approach consider the following (Table 4) 2) The “hospital diet” group subjects in the Elgin Project were fed varied diets which provided up to 10% linoleic acid in about 70 g of fat. The tocopherol in their diet (epproxi- mately 15 IU) prevented any obvious inade. quacies although an occasional increase in the peroxide hemolysis test was recorded. Their plasma tocopherols were usually above 0.6 ‘mg/100 ml. The depot fat linoleic acid of this ‘group varied from 5 to 11%. After 13 years of observation of the serum tocopherols and erythrocyte peroxide hemolysis tests of these subjects, one would have to conclude that they were on the borderline of adequacy in their vitamin E intake. 2) The first controlled diet group that was depleted was fed a diet that included 30 g of stripped lard in a total of 35 g of fat. The total diet was estimated to contain 15% linoleic acid jn the fat and less than 4 mg of d-a-tocopherol |1190 hoRwirt TABLE Vitamin E and potyisatorate fatty acids in diet and resulting linoleate in sspiated tse Ta with eseustons of requirement fom equstion” Linclste Status E Tocopherol in insewmand Mg ingested, —— depottat “peroxide fom me * 4 * Remolysis formal Hospital a2 S10 47 Sdl bordering 30g lard m5 ir adequate 30g on oll ie 3B On equate) 606 com ol 2 2M 33 Inadequate ©) 0 comm ail 3 2M 33 adequate Gog best at 3 eer 38 inadequate 60g slower 8 ‘3 ‘50 adequate @) 60 com all wos 2% 33 inadequate com oll 3 2s 3 imdequate 0.25 (& PUFA + PUFA) +4 = da-tocopherol equivalent (mg), There was no doubt about the inadequacy of the diet (2). The addition of 15 mg d-a-tocoph- erol acetate to these subjects caused an immedi- ate rise in the plasma tocopherols which decreased during the following 10 months to preexperimental levels-as -the tissues became somewhat more unsaturated. One may there- fore conclude that the total of about 17.5 mg of d-a-tocopherol (about 26 IU) which this group consumed for 20 months was adequate to protect the tissue linoleates accumulated. We were not analyzing depot fats during this part of the study. Application of the suggested equation gives a requirement of 12 mg d-a-to- copherol required. 3) In the supplemented group of subjects, when the 30 ¢ of stripped lard was replaced with 30 g of stripped com oil (3), the diet consumed provided approximately 38% of linoleates in 55 g of fats. There was an almost immediate drop in plasma tocopherol when the stripped corn oil replaced the stripped lard although stripped com oil had a bit more tocopherols than the lard. 4) Doubling the stripped com oil to 60 g caused a decrease in serum tocopherol raising a question about vitamin E adequacy. '5) Raising the d-a-tocopherol supplement to 30 mg/day for 1S months, making the total intake 33 mg of d-a-tocopherol, increased the level of plasma tocopherols slightly. 6) How the tocopherol requirement is af- fected by feeding a controlled diet of saturated fats has been shown in Fig. 2 which illustrates data from a group of male subjects who w. fed the basal diet with 60 of beef tallow the fat, These subjects remained on ths diet saturated fat for 5 years. These subjects h been fed the basal diet plus 60 commerc com oil for 6 months previously. The beet | diet (the basal constituents of the diet remain the same) was shown by analysis of composite samples to range from 2.6 to 35 linoleic acid with a mean of 3.0% in 80 g total fat as consumed. The tocopherol cont of this diet was no higher than inthe defi diets previously used, analyzing out to appro mately 3. mg dtocopherol/total daily di Despite the fow intake of vitamin E, there wa tise in serum tocopherol dung the subseque 20 months while the depot fat linoleate we rapidly decreasing. Part of this increase plasma tocopherol may have been due 10 t increase in serum lipoproteins a8 eviderc an inotease in plasma cholesiiol during d first month on she beet fat. From the data, o has the «hoi of concluding that the depot f seve releasing stored tocopherol, ch a ently did not happen during the months b beef fat was started, or that very lit tocopherol is needed when the diet conta barely enovgh linoleic acid to. supply essential fatty acid requirement, In any. ct after 20 months on the beef tallow diet marked decrease in plasma tocopherol. cor menced and this was confirmed by po peroxide hemolysis tests when-—the- fi {ocopherols dropped bstew 0.3 mT00 mlJemolyses test results rangea ieteen 50 and Jo% after the fourth year on the beet ft diet. jevertheless, these analyses showed a much ilder deficiency in these subjects than when saturated lipids had been fed as part of same sal diet and it is likely that an additional 3 3g of atocopherol/day would have~been }icient ts repair the blood tocercrol levels p this wnusuc!-emet, Recopitulating, this beef 1 diet provided 3 mg a-tocopherol in a diet hich had about 3.0% linoleic acid in the total pids and the cepot fat linoleates dropped to ss than 8%. Incidently, there were no clinical is of a fatty acid deficiency in these subjects, br did any appreciable amounts of eicosotrie- bc acid appear in ther blood. |7) Subjects on the safflower oil diet have ready been described (Fig, 1). Figure 3 selects ‘0 of these subjects chosen because they arted with markedly different levels of plasma copherol and depot fat linoleate. Both had aurked increases in plasma tocopherol during 2 first 2 years after which, as the depot fat oleates increased to levels above 30%, there ae a : : ° x10 & 20. 5 3 2 20 Pe co 7 ly [og 3 : oe 4 ‘62 63 64 65 66 3. 3. Shows two of the subjects from Fig. 1 10 Bie individual variations, Note thatthe leaner of Wo had the more rapid change in depot fat te Revels 35 well a5 the lrgor decrease in plasma hherol when his linoleate concentration surpassed {the foul faty acids in the depot HUMAN REQUIREMENTS FOR VITAMIN E ust was @ significant decrease in plasma tocopherol which continued to fall as the depot fat linoleates increased. The weights of the subjects femained coastzat but note that the thin subject-(Bi3) who started at a lower level of aépot fat linoleate had 2 more rapid increase in his adipose linoleate. Since these subjects were consuming 25 mg d-a-tocopherol in their food per day, the decrease in plasma tocopherol was unexpected. One would have expected such a large daily intake to be reflected in higher plasma tocopherol levels in the blood drawn the following morning, Unfortunately, these experi- ments ended and we did not lear the consequences of decreasing the tocopherol intake in subjects with such high levels of ‘adipose linoleate. Even higher levels of antioxi- dant may be required under these unusual conditions. One wonders how much higher the plasma tocopherol should bein order to achieve the greater protection that may be needed when the tissues have such unnatural levels of PUFA. 8) The last 2 lines in Table 4, ae'calculated from the depleted subjects who were supple- mented with d-a-tocopherol and described in the frst halfs of Table I and Table 2. Note that 10.5 mg and 18.5 mg of d-a-tocopherol in a diet in which the fat contains 42% linoleic acid are inadequate. ‘The data in Table 4 make it difficult to change our original decision that the adult male vitamin E requirement ranges from a minimum of 10 to a piobable maximum of 30 mg of d-actocopherol/day despite the fact that many individuals in apparent good health consume considerably less. Although 15 TU/day appears to be adequate for most individuels fed the avarage American diet, if I am to believe in the experimental method, the data show that 15 IU, have litle, if any, safety factor. The only point in favor of a 15 TU requirement is that so many of us are apparently healthy consuming ‘no more tocopherol. Is the effect of borderline tocopherol inadequacies, one which causes tissue deteriorations too slight to be measured by techniques currently available? For example, we have no method of determining a 3% increase in the rate of erythrocyte or other cell turnover. For one who has preached against the need for tocopherol supplementation this is a soubsearching question. Fortunately for my conscience, unlike the other fat-soluble vita-92 | mins, large amounts of tocopherol are not toxic. ‘Summary 1) Evaluations of the human requirements for vitamin E have been reviewed, Reinterpreta- tion of the data from long-term studies have made use of information on changes occurring in the linoleic acid content of depot fats in j relation to plasma levels of the tocopherols. 2) Three variables should be considered in evaluating the requirement for vitamin E: fist there is a need related to the synthesis and incorporation of the large amounts of PUFA into vital compounds even when the amount ‘of PUFA in the diet is low; second, an allowance should be made for the percentage of PUFA in the dietary fatty acids; and thicd, an allowance should be made for the amount of such PUFA consumed 3) The calculation suggested is relatively simple. A basal minimum of 4 mg da-tocoph: erol is allowed for nonnal synthesis and accumulations of PUFA into phospholipid, lipoproteins and other compounds even when the amount of PUFA in the diet is ery low. To the basal minimum is added an amount of datocophetol equivalent that is equal to 0.25 times the sum ofthe percentage of PUFA in the dietary fats and the amount in grams of the PUFA consumed. The results of using Such a calculation compare well with data from experimental studies of the vitamin E require- rents of adult men. 4) In effect, the range of 10 to 30 mg of d-atocopherol formerly suggested as the adult requirement is reinforced. Any table of vitamin E requirements should clearly demonstrate this range. References 1. HARRIS, P. L, AND N. D. EMBREE. Quantts tive consideration of the effects of PUFA content fof the diet upon the requizement of vitamin E ‘Am. J. Clin, Nutr, 13: 385, 1963, 2. HORWITT, M. KC. 'C. HARVEY, G. D. DUNCAN "AND W. C. HARVEY. Effects of Timited tocopherot intake in man with relation- ship to erythrocyte hemolysis and lipid oxida tions. Am. J, Clin. Nur. 4: 408, 1956. 3. HORWITT, MK. Vitamin E and lipid metabolism in man, Am. J. Clin, Nutr 8: 451, 1960, 4. HORWITT, MK. lntericlations between vitamin E and polyunsaturated fatty acids in adult men, ‘Vitamins Hormones 20: $41, 1962. noRWwiTT 5. BAKER, E. M., H. E, SAURERLICH. § WOFSKILL, W. 7. WALLACE AND E. E.DE Tracer studles of vitamin C utilization in n Proc. Soc. Exptl Biol. Med, 109: 737, 1962. 6. HUME, E. M., AND H. A. KREBS. Vitumb quirement of human adults. Med. Res. Cou Spec. Rept. Ser. No. 264, London. 7. HORWITT, M.'K., E, LIEBERT, 0, KREISL AND P. WITTMAN. Investigations of hur requirements of B-complex vitamins, Nat. Ac Sci-Natl, Res, Council, Bull 116, Washingt DG, 1948, 8. HORWITT, M. K,, C. C. HARVEY, W. ROTHWELL, J.) CUTLER AND B. ti) FRON. Tryptophan-niacin relationships in man Note. 60: Suppl. 1, 1956. 9. QUAIFE, ML, 'W. J. SWANSON, M. Y. D AND P.'L. HARRIS. Vitamin E in foods a tissues. Ann. N.Y. Acad. Se. 52: 300, 1949. HARRIS, P. L,, AND N. D. EMBREE, Quant tive consideration of the effect of polyuns rated fatty acid ‘content of the dict up fequirement for vitamin E. Am. J. Clin. Nut 385, 1963. AL. HORWITT, M. KC. C. HARVEY AND E. HARMON, Lipids, a-tocopherol and erythroc hemolysis. Vitamins Hormones 26:487, 1968. BIERI, J. G., AND R. P. EVARTS. Gam tocopherol: metabolism, biologleal activity. a significance in human vitamin E nutrition, Am Clin. Nutr, 27: 980, 1974 ). HARRIS, P. L. Practical nutritional aspects vitamin E. Ann. N.Y. Acad, Sci 52: 240, 1949, |. HORWITT, M. K. Tocopherol and polyunsa ated lipi’ relationships. In: International Sy posium on Vitamin E, edited by N. Shimazo 4nd Y. Takagi. Tokyo: Kyoritzu Shuppan C Led, 1972, p45, HORWITT, M. K., C. C. HARVEY, CH. DAH IR, AND M, T. SEARCEY. Relationship be Tocopherol and serum lipid levels for the det ‘mination of nutritional adequacy. Ann. ‘Read. Sei 203! 223, 1972, HORWITT, M._K,B. CENTURY AND A. ZEMAN. Erythrocjte survival time and retic fyte levels after tocopherol depletion in man. 3. clin Nusr. 12:99, 1963, . CENTURY, B., AND M. K. HORWITT. Role diet lipids in the appearance of dystrophy a txeatinurla in the vitamin E-deticent rat. 3.N. 72: 357, 1960. WITTING, L.'A., AND M. K. HORWITT. Fit of degree of fatty acid unsaturation in tocophed eficiency induced creatinutia, J. Nutt. 8 1964 - 19. CENTURY. B., 1_A-WITTING, C. C. HARVI AND M. K. HORWITT. nterelationships ictary lipids upon fatty ackd composition brain, mitochondria, erythrocytes and heart ts in chicks, Am. J. Clin, Now. 13: 362, 1963. FITCH, C. D., J. S. DINNING, L. A, WITTIN AND M. K. HORWITT. Inftuence of dietary ‘on the fatty acid composition of monkey erytM cytes. J. Note. 25-341, 196), 2. 16. 1B. 20,— HUSAR REyoORBMENTS. FOR ViraMne-E- and metabolism, Arch, Inirrat Med. 105: 33, 1960. smmittee on histvecal Handbooks. Metabolism, by PL. “Aluman and D. §. Dittmer, Bethesdv, Sid: Federation Am. Soe. Exptl Biol, 71966, £5 Discussion R.A. Witting: I think “lack Bieri’ figures show fiat’ we have been consuming a lot more jybean oil, and T very much doubt that we ike in very much of this without prior pydrogenation. I would think that the trend in ocessing is tO remove more polyunsaturated stty acid than to remove vitamin E. Would you are to comment on that? (. K. Horwitt: No, but that is not pertinent to ty formula because Iam interested only in sating requirement to consumption of polyun- iturates, G. Bieri: 1 might just comment that 60% of le soybean oil consumed in the country goes to margarine and shortening 50 2 lot of it is rdrogenated. Two years ago, when I got orwitt’s opinion on revising the RDA's, he 4 of the opinion that they might be revised ywnward. Today I think he just showed he is ling to stick with the old figures. I might say, f those that are interested in the RDA's, that # interpretation of the new edition is that 2y are not going to be changed drastically, Lis 23, WATT, B. K., AND A. L. MERRILL. Compost tion of Foods. Agric. Handbook No. 8, US. Dept. of Agriculture, Washington, D.C. 1963. 24, Nultitive Valie of Foods. US. Dept. of Agricul ture, Home and Garden Bull, No. 72, Washington, D.C, 1970, although 1 think I should be honest and confess that I think we are still leaving the user somewhat in the dark as to what the RDA is. The difference is that previously the figure that appeared in the table was the maximum figure intended for the person who is eating high levels of PUPA. This was very misleading in my pinion and in’ the opinion of the committee, This time we are putting in the table the figure which we think represents what most average diets have in terms of PUFA but we are stating in a footnote “see the text” where we clearly state that this figure can vary. : M. K. Horwitt: 1 think you are quite right in noting that about a year ago I had left the impression I would be satisied with 15 mg for average consumption, but also, at that time, 1 stid we should have a footnote such as you described to cover increased consumptions of PUFA. My more recent extensive calculations make me feel less certain about accepting 15 1U as an average level. After all, this is only about 10 mg of a-tocopherol a day.