E. F. KOHMAN1 Campbell Soup Company, Camden, N. J.

(Received for publication

April 25, 1939)

Downloaded from jn.nutrition.org by on July 31, 2007

One is apt to gather from nutritional literature that vegetables do not supply calcium efficiently. Because this is not in accord with the results of a number of experiments, some covering several generations of animals (Kohman, Eddy and associates, '31, '34), this discrepancy presented itself as a problem wanting solution. As there seemed to be con fusion about the amount of oxalates in foods and a scarcity of data as to the behavior and fate of oxalates in the diet, information on these points might be expected to throw light on the availability of calcium. It seemed desirable for higher accuracy, to make oxalate determinations under conditions that would avoid drastic treatment of the food and entail the concentration of the oxalate of a relatively large sample, freed from most of the other food constituents.

The following procedure was used. The samples, 400 gm., were first cooked tender by steaming without drainage loss. After thorough disintegration and an approximation of water content, sufficient concentrated hydrochloric acid was added to yield a 15% solution in the water present. This was
1The experimental work embodied in this paper was done in 1934-1936 while the author was with the Research Laboratory of the National Canner 's Association, Washington, D. C. There he had the assistance of N. H. Sanborn and D. C. Smith. This opportunity is taken to acknowledge their part in the work. 233

poke. The sample was then pressed in cheese cloth and the weights of the drained material and of the residue recorded. after filtration if necessary. This as sumption was proved correct for pineapples and turnip greens by adding varying amounts of oxalic acid and subsequent analysis. notably spinach. All determinations were made in duplicate and the average is given in table 1. expressed . After wash ing and redissolving a second. all the results were averaged and the number of samples is given in parenthesis. Murray and Watkins ('25) and suitably proportioned for this purpose using sintered glass to disperse the ether. Swiss chard. F. THE OCCURRENCE OP OXALATES IN FOODS Downloaded from jn. per minute.cm. The precipitate was then titrated with permanganate.234 E. the ether evaporated and the water solution filtered. New Zealand spinach. deep and the ether coursed up through it from the 2. Calcium oxalate was then precipitated.org by on July 31. To study varietal and other factors more than one sample of some products were analyzed. not less than 2 days. A 250 to 300 cc. By direct titration of a highly diluted sample. In this the liquid to be extracted was 32 to 37 cm. KOHMAN allowed to stand at room temperature. beet tops. and if necessary. lamb's quarter.nutrition. of fritted glass at a rate of 20 cc. By precipitating the calcium in the titrated medium as calcium oxalate. the chlorine in the two fractions was determined. In them. purslane and rhubarb have a high oxalate content. with occasional stir ring. As no significance could be placed on the differences. the purity of the original oxalate could be gauged. water was added to the extract. On the assumption that the oxalic acid distributed itself in the same ratio as the chlorine. a third pre cipitation was made for complete purification. 2007 Only a few foods.6 sq. sample of the drained liquid was subjected to 24 hours of wet ether extraction in an apparatus modeled after that suggested by Palkin. it could be calculated for the entire sample by a determination on the drained fraction. When the extraction was complete.

059. sweet. stalks Chard. sweet. lima (2) Beets.org by on July 31.TABLE 1 Oxalic acid (anhydrous) in foods Product Total solidi Oxalic acid Vegetables Asparagus 6. 2007 THE JOURNAL OF NUTRITION. leaves Purslane. Chinese Carrots Cauliflower Celery stalks. Irish Potatoes.904. NO.028. land.606. yellow (2) Cress.5814. white Corn. green pod (3) Beans. Swiss.6610. soup leaves Celery.6610. Swiss. VOL. 3 . green Parsley Parsnips Peas Peppers.759.4511.526.477.587. wild Cress.258. stalks Cabbage (2) Cabbage sprouts Cabbage. 18.nutrition. stalks Radishes 235 Downloaded from jn. unpeeled Beet leaves Beet stems Broccoli.507. sweet. green (3) Poke Potatoes.2012. Swiss. wax Beans. leaves and flowers Broccoli. Beans. minus leaf ribs Lettuce (6) Mustard greens (3) Okra Onions.236.808.8024. bleached Celery. leaves and stalks (2) Chenepodium (Lamb's quarter) Corn. leaves Chard. early fine curled Cucumbers Dandelions (3) Egg plant Endive (5) Escarole Kale Kale.578. sweet Purslane. soup stems Collards (2) Chard.

early summer Apricots Avacados Bananas Berries. Bartlett Pineapples.6210. cantaloup Melons. Bing Currants. New Zealand. green summer Turnips.8122.826.nutrition.6214. peel Limes. water Nectarines Oranges. canned (12) Spinach. leaves Spinach.6023.157. peel Mangoes Melons.2520.TABLE 1—Continued Product Total solidi Oxalic acid Calcium Vegetables Rape Ehubarb Spinach (53) Spinach. 2007 .1013. black rasp Berries. houey dew Melons. edible portion Oranges. New Zealand.265. red rasp Berries. sweet. red Grapes. green goose Berries. stalks Squash.org by on July 31. black Berries.518. Italian Tomatoes 10. casaba Melons. damson Plums. Hawaiian canned Plums. dew Berries.2512. blue Berries.5813. green gage Prunes.588. peeled Turnips. Concord Grapes. Hiley Pears. peel Peaches. Thompson's seedless Grapefruit Lemons. red sour Cherries.166.357. Alberta Peaches. juice Limes. unpeeled Turnip greens Fruits Apples. straw Cherries. Downloaded from jn. juice Lemons.608.8112.

a basal diet of one can each of roast beef.5).4740. including practically all commercial and many experimental varieties grown in California and in Maryland as well as those shipped from Texas.nutrition. minimum 0. peas. It is notable. uniform supply for .44) while Maryland spinach averaged 1. that oxalate was obtained from all but a very few products. The essential analysis expressed in grams was : canBoast One weight362607599551Solids13311145184Calcium0.133Phosphorus0.25% (maximum 2. it was possible to have a constant. This hope was not realized. California spinach averaged 0. California spinach was only slightly lower in oxalic acid but markedly lower in calcium.1470. such a variation raises a number of important questions. One purpose of the analytical survey was to ascertain if some varieties of spinach might be relatively free of oxalate. In fifty-three samples. Eddy and White ('37) used diets of canned foods and found them to supply calcium efficiently.251 beefPeasCarrotsSweet potatoesNet By securing a sufficient amount from one lot of each of these. In previous experiments Kohman.0540.50.1550.1560.02% on the dry basis (maximum 12. PLAN OF FEEDING EXPEBIMENT8 Downloaded from jn. the average anhydrous oxalic acid content was 9.4% of the usual crystalline form of oxalic acid.92% (maxi mum 2. 2007 As a number of widely used greens contain insignificant amounts of oxalates. however.42).6.44). minimum 0.1060. For convenience. mostly fruits. Green leaf-vegetables are our richest calcium sources.59% calcium (maximum 0. it is often considerably over 10% on a dry basis. One per cent in table 1 is equivalent to 1. Since a considerable number of mineral elements are dietary essentials. Florida and Carolina. The calcium values averaged 1. In most foods there are present mere traces. a comparison of these with spinach seemed in order. therefore.50.OXALIC ACID IN FOODS 237 as anhydrous oxalic acid.84. carrots and sweet potatoes was chosen.org by on July 31. minimum 4. minimum 1.

it decidedly interferes with both growth and bone formation.3%. which was half the amount of spinach added in diet B to supply 60% of the calcium.8% of the calcium. It was added in that amount to be compared with diet D. This cannot be ex- . 2007 It is apparent from data in tables 2. Except as noted otherwise the duration of the experiments was 21 days. 3 and 4 that whereas spinach greatly increases the calcium content of the low calcium but well performing basal diet. The rats used were purchased from a large dealer who was able to supply large litters suitable for apportioning between the diets of a given experiment equally as to weight and sex.22%. evincing efficient calcium.238 E. 0. Diet E is equal to D both in fiber and calcium. F. Small additions of greens (5 to 8%) were made to supply 60% of the calcium of the final mixture. i. three mesh to the inch. These four foods.nutrition. some additions to the basal diet were on a different basis. in the above proportion. liquid and all. were thoroughly mashed and mixed. The two greens in diet D supplied 70% of the calcium and raised the content to 0.093%. RESULTS OF FEEDING EXPERIMENTS Downloaded from jn.org by on July 31. There was an oc casional death during this period and in every case it was an animal receiving either spinach or calcium oxalate. Six animals were always placed on each diet and kept in one cage with a raised wire bottom. KOHMAN an entire series of experiments conducted over an extended period of time. The turnip greens in diet C sup plied only 58. This raised the calcium content to 0. Also in experiment 3. in which it was accompanied by an equal amount of spinach. In experiment 3. since the effect of crude fiber on calcium availability has often been questioned.. This diet was obviously low in calcium. diet D was planned to be equal to diets B and E on the basis of crude fiber. table 2.e. They were started on the experiments at 21 or 22 days of age when they weighed 30 to 35 gm. It permitted good but not maxi mum growth and bone formation. and hence had to be eaten in that proportion.

24 40Diet B gm.345.144.24 gm.939.1Experiment daysCalcium 6— 21 carbonateSpinach.340.745.)Turnip (8. 0.047.62 87DietC 74Diet D gm.750. spinachSpinach + CaCO.340.org by on July 31.045.2Experiment daysNoneSpinach 3— 21 54Diet A gm.553.440. 2007 1Canned spinach brand 2Canned spinach brand 3Freshly spinach brand cooked spinach534758576245.137.5%)Spinach greens (8.846.541.2%)yrn.4%-11.549.62 gm.943. + spinach 11.4Experiment daysNoneCalcium 2— 28 carbonateCalcium oxalateTurnip (4.651.1 .233.153. -li in daysTurnip 1— 21 (4.8%-23.553.5Experiment E daysNoneCanned 4— 21 Downloaded from jn.Spinach greens 11.406 gm. equivalent to oxalate in (7%)Kale (6%)Mustard greens (5%)768857798451.147.)Turnip greens (4.5%)Spinach greens (8.5938%43.645.2%)6794791067744.2Experiment daysNoneCanned drainedCanned spinach drainedCanned spinach drainedCanned spinach drainedFreshly spinach (9%)Freshly cooked cooked 7— 21 brand 1— not brand 1— brand 3— not brand 3— spinach — drained not spinach — drained (7%)5245545256547547.OXALIC ACID IN FOODS 239 TABLE 2 Growth of rats and bone ash of alcohol-ether extracted dry tibia Dietary additionWeight tibiaExperiment gainA.853.Diet 4644.62 23.nutrition. + CaCO.

the food was removed the evening before. 16 hours. the difference between 79 and 38. from two to nine times as much calcium as the spinach contains.1% oxalic acid and 1. Table 3 records the gain in weight and the gain in calcium per animal in other similar feeding experiments. table 3. and bearing in mind that 15% of the total calcium is only 38% of that supplied by the basal diet. kale. the food consumption was determined and from this the calcium utilization obtained.e.nutrition. Each figure represents the average of six animals. markedly improve growth and bone formation under similar conditions.8% oxalic acid. 2007 . To arrive at the calcium per animal at the time the feeding was started. This would require. It appears that when turnip greens were added to the basal diet 79% of the calcium was utilized but when spinach was added only 15% was utilized. Assuming that the calcium supplied by the basal diet and the turnip greens was equally available. In experiment 8.240 E. F. such as sleekness of fur. In table 2 are recorded the results of a series of experi ments in which the gain in weight and the per cent ash in the tibia was determined after feeding the basal diet alone and also the basal diet with various additions.e. The sample of spinach used had 10. but it actually rendered unavailable 41% of the calcium of the basal diet. A general impression was obtained that some superiority.org by on July 31. In all cases when an entire animal was analyzed. such as turnip greens. performance in the rat is then comparable with other greens. a similar group of six animals was sacrificed and their calcium content determined. When calcium oxalate was the addition. The diet to which it was added then had 0. KOHMAN plained on the basis of crude fiber. to similar groups of animals. 44% of the calcium was Downloaded from jn. resulted when a low oxalate bearing green supplied calcium as against an equivalent addition of calcium carbonate. it appears that the spinach not only supplied no available calcium.. On the other hand greens with negligible oxalate content.53% calcium. i. i. If enough calcium carbonate is added with the spinach to balance stoichiometrically its oxalate. in the fifty-three samples analyzed. mustard greens and collards.

TABLE 3 Growth record and gain in calcium in Addition to basal diet(iiiin weightCalcium gainExperiment Total per rat Net gain per gram gain utilized%——70441579 in weightgm.2449 oxalateSpinach Calcium (8.OXALIC ACID IN FOODS 241 utilized.0596 0. indicate a superiority of turnip greens over calcium carbonate in growth.20330. showing that the rat can make use of a small portion of the calcium in it.004100.3447days0.002860. indicating that the ability to oxidize it increases with age.42050.0. The collection period was 7 or 8 days duration.16820.002670.00300Calcium ACID IN URINE AND FECES OF CALCIUM AND OXALIC In a number of these experiments the calcium and oxalate excreted in the urine and feces were determined in addition to the calcium and oxalate in the food consumed and the calcium deposited in the tissue.0275 0. The collection was begun 3 or 4 days after the animals were placed on a diet.00074 Turnip greens (4. The data in tables 3 and 4.00106 Downloaded from jn.5%)ffm. It is notable that with turnip greens in the diet four times as much calcium per gram of tissue was deposited as with spinach in the diet. experiment 8.001990.nutrition. thus allowing two periods in a 21-day experiment. 2007 21Controls 5— — days old 21 0.35350. calcium utilization and calcium deposition per unit body weight.2%) 0. .0.11150.5300Experiment 21Controls 8— — days old 21 carbonateCalcium Calcium oxalateSpinach Turnip greens715637810.625684ffm.4603daysffm.2172 RECOVERY 0.0. Also there is better utilization of calcium in the second period.1853 0. There seems to be a tendency for less oxalic acid recovery in the second period.32860. This included any endogenous oxalic acid which would be overshadowed by the spinach oxalate.2447 0.2431Calcium 0.org by on July 31. The data are recorded in table 4.

. The exTABLE 4 Secovery of oxalic acid and calcium Dietary additionPer cent oxalic acid recovered In In T t j TotolExperiment urine fecesPer cent calcium Calcium recovered utilized In In urine feces nvj. C Period 1 Spinach \ .org by on July 31. . ... 13 ( Period Spinach not drained < .nutrition. it represents a relatively high percentage of as similated calcium.( 2 344157Experiment 3465332 42 11865023572763 ( Period 1 Spinach drained f _. In only one case with spinach in the diet was as much of the calcium utilized as was supplied by the basal diet. probably passing as oxalate through the excretory organs.. 2-. Period. 2007 81038 628 626 7018974 3082 91 . KOHMAN While urinary calcium may not seem high with spinach feeding. . ( Period 21 386 8( Period Calcium carbonate •}_.242 E. 12 84 96 ( Period 214 90Experiment 78 12 5. .1 2Calcium oxalate Spinach81039234733*Experiment 3. . 77 91 f Period 1 Spinach X 2Spinach I Period 18 72 90 + CaCO3 Õ_e"°.. . f Period 1 Calcium oxalate <—_. [ Period . _ 2Spinach ( Period \f Period 1 Turnip ereens •! ( Period 213 93 611855 7333 2468 7944 3218 33 643 6373 1546 6676 8554 3424 62574088 66594312 445341103603370363064Experiment 106655 34473469 Downloaded from jn. F._ J Period 214 7..„ 1 2( ¿ Penod Period 1 Calcium oxalate < _. .

The wide un calcified area in figure 2. There was.org by on July 31. very poor calcium utilization even with drained spinach. In this the spinach was cooked by dropping it in twice its weight of boiling water. In due time two litters of nine young each appeared on the diet with turnip greens.OXALIC ACID IN FOODS 243 ception is in table 4. 2007 EXTENDED FEEDING PERIOD In one experiment twelve animals at 21 days of age were placed on each of two diets. Figures 1. Some time later one litter appeared among the animals on the diet with spinach. with spinach added and with turnip greens. its weight was 58% of the original. In one group spinach supplied 60% of the calcium and in the other turnip greens. Downloaded from jn.e.. The undrained spinach was cooked 20 minutes in its adhering water. all but two of which were dead and these were shortly eaten by their mother. while the average weight of their parents at the same age was 26 gm. Spinach calcium is insoluble and hence not extractable. experiment 7. After draining. The band across which the line is drawn represents the dentine layer of a rat's tooth magnified 310 times. the basal diet alone. representing the spinach diet is . in one litter and 34 in the other. five animals on the spinach diet had died while all those on the diet containing turnip greens were in excellent condition. These were reared to the age of 21 days when the average weight was 39 gm. By the time the age of 90 days was reached. table 2.nutrition. however. Numerous experimental and com mercial blanching tests showed it is difficult to remove more than half the oxalate by this process. while of those receiving turnip greens it was 205. i. allowing 7 minutes to return to boiling and then boiling 10 minutes. 2 and 3 are photomicrographs of a tooth of an animal respectively on each of the three diets in experiment 3. The dark portion of this band designated 'A' is calci fied while the light portion 'B' is uncalcified and represents the area where new dentine is being formed. The average weight at 90 days of age of animals receiving spinach was 134 gm.

New Zealand spinach. a high percentage of deaths occurs among rats fed between the age of 21 and 90 days. tooth structure is disorganized and dentine poorly calcified. DISCUSSION Insofar as these data are on a comparable basis they are in agreement with the reports of Tisdall and Drake ('38).org by on July 31. Reproduction is impossible. peas. . Some oxalic acid is assimilated and tends to carry calcium with it into the urine. expressed as anhydrous oxalic acid. lamb's quarter. have been shown to occur to the extent of about 10% on a dry basis in spinach. In view of the low solubility of calcium oxalate. If to a diet of meat. beet tops. It should be mentioned here that the bones of animals receiving spinach were very soft and pliable.244 E. They all dealt with calcium availability. rela tively low in calcium but permitting good though not maximum growth and bone formation. Spinach not only supplies no avail able calcium but renders unavailable considerable of that of the other foods. KOHMAN apparent. Swiss chard. Fairbanks and Mitchell ('38) and Fincke and Sherman ('35). No one seems to have studied hereto fore the fate of the oxalates in the diet of the rat. The bones are ex tremely low in calcium. F. much more in the feces. purslane and rhubarb. 2007 Oxalates. poke. This is perhaps of equal importance. carrots and sweet potatoes. Considerable of the oxalate appears in the urine.nutrition. spinach is added to the extent of about 8% to supply 60% of the calcium. there arises the question of any possible damage of its passage through the excretory tissues. The demonstration of almost universal presence of small amounts of oxalates in vegetables and fruits tends to throw doubt on the quantitative aspects of endogenous oxalic acid. Traces were found in nearly all vegetables and fruits. published since this work was done. CONCLUSION Downloaded from jn.

.i — s %% 245 P* £3 o s .CM .Vi £ a .• . • .org by on July 31.*. 2007 P nm 3 ^» «kZ l-: CO t--.nutrition.~ CD M Downloaded from jn. x = ^ ** I hi O t...

S.nutrition.. H. New York.. E. Eng. J. W. KOHMAN Turnip greens. Chem. Dental Corps. Beltsville. L. FINCHE. Eng. Nutrition. 1064. vol. Acknowledgment is made to Capt. 612. 16. H. KOHMAN. U. 758. W. vol 110. 16.. F. EDDYANDCELIAZALL GÃœRIN1931 A canned food diet. in skim milk powder and in calcium oxalate. H. S.W. p. WHITE 1937 Comparative ex periments with canned.246 E. F. J. under similar conditions produce excellent animals that deposit four times as much calcium per unit body weight as those receiving spinach.. MURRAYAND H. E. Ind. J. Nutrition. where various samples were obtained. ANDH. Irons. 613.. KOHMAN. H. California.. TISDALL. F. F. 26. AND H. R. Ind. H. Ind. p. EDDY AND CELIA ZALL GURIN 1934 Calcium and vitamin D. LITERATURE CITED Downloaded from jn. DRAKE 1938 The utilization of calcium. Chem. who prepared the photomicrographs. 23. .. vol. F. M.org by on July 31. 9. 14. Biol. p.. SHERMAN 1935 The availability of calcium from some typical foods. L. W. N. mustard greens. vol. J. AND T. and to the experiment stations at Davis..B. SANBORN.. EDDY AND MARY E. F. WATKINS 1925 Automatic devices for extracting alkaloidal solutions. A. 2007 FAIRBANKS. G. Maryland and Geneva. KOHMAN. G. home cooked and raw food diets. H. greens with negligible oxalates. p. Chem. Nutrition. p. vol. p.E.. PALKIN. MITCHELL 1938 The availability of calcium in spinach. kale and collards. vol. A. A. Eng. 421. 17. C.. 79. Chem. p. vol.

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