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VITAMIN C CONTENT OF VEGETABLES. I. SPINACH”

D. K. TRESSI,ER, G. L. MACK, AND C. G. HIKG

New York State Agricultural Experiment Station, Geneva, New Pork, alzd the Department of Chemistry, University of Pittsburgh, Pittsbwgh, PennsyEva.nia

(Received for publication, October 23, 1935)

Fresh spinach has long been known to contain relatively large amounts of vitamin C ; however, the amounts reported by the several workers differ greatIy. One difficulty in comparing the published results lies jn the fact that there is littIe uniformity in the mode of re- porting the data obtained. Most analyses reported recently have in- dicated the amounts of ascorbic acid in milligrams per gram of spinach. If one assumes, as suggested by Bessey and King (1933), that

0.5 mg. of a.scorbic acid per day is required for protection of a guinea

pig from scurvy, the data in recent papers may be recalculated on a uniform basis (Table I.).

 

TABLE

1

Previous Reports OR the Asambic Acid Content of Spinach

 

Investigator

Source or rariety of spinach

Ascorbic acid

 

mg. per gm.

Eddy, Kohman, and Carlsson

“Market”

 

>O.G

(1925)

Hessler and Craig (1929)

“Fresh”

0.07

to

0.10

Hessler, Johnson, and Walsh

Bloomsdale

 

0.17

to

0.25

(1931)

Von liahn (1931)

Unknown

 

0.10

Wasson (1931)

“Fresh

<O.B

Kifer and Munsell (1932)

Virginia Sarog

0.5

Kifer and Munsell (1932)

Viroflay

 

<0.5

Kifer and MunseU (1932)

Princess JuIiana

<0.5

 

Bessey and King (1933)

Fresh

0.50

to

0.62

Bessey and King (1933)

Market”

 

0.4

to

0.5

Tauber and Kleiner (1935)

Fresh”

0.43

to 0.46

While variations in technique employed in making the vitamin C estimation may account for some of the differences in the results ob-

tained, yet it is probable that the spinach examined by the different workers did vary widely in its ascorbic acid content. As shown in data

presented above, Bessey and King

(1933), have indicated that fresh

* Approved by the Director of

the New Pork State Agricultural Experiment

Station for publication as Journal Paper No. 101, July 25, 1935.

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4

D. K. TRESSLER, G. L. MACK, AND C. G. KING

spinach is higher in ascorbic acid than “market” spinach. Wasson (1931) working in Brookings, South Dakota, states “The spinach grown in the college gardens does not seem to have as high vitamin C value as spinach that has been tested in some other laboratories.” Moreover, she also finds that “home grown spinach had greater anti- scorbutic potency than the spinach bought in the market.’ Kifer and Miinsell (1932) reported slight varietal difference in vitamin C content among three varieties tested, Virginia Savoy, Viro- flay, and Princess Juliana, the last being lowest in potency. Ton Hahn and Gorbing (1933) found that the vitamin C content of spinach varies depending on the type of fertilizer used. Unbalanced fertilizers (i. e, those especially high in some one component) yielded’ spinach relativeIy low in vitamin C, whereas soil treated with balanced fertilizers produced spinach high in this vitamin. The work reported in this paper was undertaken in order to obtain some data concerning the relative importance of variety, freshness, maturity, and soil on the ascorbic acid content of this vegetable.

METHODS USED

The method of determining ascorbic acid described by Bessey and King (1933) mas employed, but €or purposes of comparison biological assays were also made on two samples. The procedure employed in making them is described below :

Samples of the freshly picked spinach were carefuIly packed, frozen, and kept in a container with dry-ice. The curative type of test was used as a measure of antiscorbutic value. The guinea pigs were kept on the standard Sherman and Smith (1931) diet supplemented with a generow allowance of fresh spinach for two weeks to assure their normal. growth rate, and to permit them to reach approxi- mately 325 gm. in weight. They were then given the basal diet only for 14 days, at which time there was evident an average slight loss in weight. Test feeding mas then begun with weighed port.ions of food supplied separately to each animal, The quantity to be fed was calculated from the indophenol titration value. The food was consumed rapidly so that there would not have been a significant loss from oxidative destruction before it was eaten. Control groups mere given stan- dard portions of freshly prepared vhmin solution, fed from graduated pipettes. It is evident from the data (Table 2) that the animal assays correspond closely with the indophenol titratiou values.

EXPERIMENTAL

Relatz/ue Potency of Leaf aid fitern

Appreciating the fact that leaves and leaf petioles often differ markedly in chemical composition and therefore might contain very different quantities of ascorbic acid, the ascorbic acid content of leaves and stems of freshly harvested spinach grown on Ontario clay loam soil was determined. The ascorbic acid content of the leaves and

VITAMIN C CONTENT OF VEGETABLES

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stems of the Prickly Winter spinach was found to be 0.78

and 0.06

mg. per gram respectively; that of the Ieaves and stems of

the Hol-

landia variety was 0.79 and 0.04 mg. per gram respectively.

These

data indicate that whereas the leaves are high in ascorbic acid, the

sterns are almost devoid of this vitamin. This shows that

in euam-

ining spinach, either the entire plants should be taken or

else

the

TABLE P

Biological Assay of Yitamin G in S@ncbeh LFfiWe8

 

Scu rr?

Basal diet supplement

No. of

animals

Vitamin C

fed per day

Initial

weight

Gain in weight

during 14 days

scare at

autopsy

w.

gni .

gm .

Prickly Winter

Spinach

  • 4 320

0.5

38

1

 
  • 4 1 .o

319

17

0

Holitlndia

Spinach

  • 4 0.5

307

35

2

309

  • 4 43

1.0

0

Ascorbic Acid Solution

  • 7 0.5

311

41

2

  • 6 1.0

310

63

0

llnsupplemented

  • 6 -113

0.0

329

19

leaires and stems should be assayed separately; otherwise it would be difficult to obtain uniformly reliable data. Since in spinach the ascor- bic acid is principally in the leaves, the data presented in this paper refer to the composition of the leaves.

Maturity and Ascorbic Acid Content

Spinach is a crop which grows and matures very rapidly in the spring. Often within one week from the time that the first leaves are ready to harvest, the crop passes optimum maturity and begins to bolt (go to seed). To determine the influence of maturity of the crop

upon its ascorbic acid content, freshly cut spinach leaves from two

varieties mere analyzed before the spinach became large enough €or harvesting, again four days later when it had nearIy attained full growth, and again when it began to bolt. On these dates the ascorbic acid content of the Prickly Winter variety mas 0.72, 0.75, and 0.78 mg. per gram respectively ; that of the Hollandia variety was 0.78, 0.75, and 0.79 mg. per gram resp’ectively. These figures indicate that the ascorbic acid content of spring spinach leaves is nearly constant during the growth period.

Influence of Variety and Soil

Twelve varieties of spinach at optimum maturity were harvested from upland soil (Ontario day loam at Geneva, N. Y.) and the same varieties harvested on the same day from muck soiI at Holcomb, N. Y. One sample from each lot was analyzed for ascorbic acid. While there

  • 6 D. K. TRESSLER, G. L. MACK, AND C. G. RING

is a varietal difference, the soil on which the crop is grown (Table 3) apparently has a somewhat more important influence on the ascorbic acid content of the leaves. It is noteworthy that on both soils the Princess Juliana variety is the lowest in ascorbic acid and that variety is the one found by Kifer and Munsell (1932) to possess the least antiscorbutic acid value of the three varieties examined by them.

TABLE 3

Ascorbic Acid Content of the Leaves of Smeral Varieties of Spring S@m&

 

&own

on Nw& and Upland Soil aad Harvested on the Same

Day

 

Ascorbic acid in leaves

harvested

 

Variety of spinach

from

 

Mucksoil

I

Uplandsoil

mg. par om.

mg. per gm.

Eskimo ....................................................................

0.62

0.77

Old Dominion..........................................................

0.56

0.70

Nobel .....................................................................

..

0.55

0.79

Vietoria ...................................................................

0.55

0.66

Viroflay ...................................................................

0.53

0.78

Broad FIanders ......................................................

0.49

0.89

Long Standing Bloornsdale...................................

0.49

0.74

Virginia 6;cvoy .......................................................

0.48

0.80

Prickly Winter .......................................................

0.46

0.75

Hollandia ................................................................

0.42

0.75

King of Denmark ....................................................

0.40

0.64

Prineess Juliana .....................................................

0.38

0.53

Mean..................................................................

0.49

0.75

On the average the amount of ascorbic acid in the spinach leaves grown on the upland soil was 50 per cent higher than that of the spinach from the muck soil. This point will be checked by examining spinach grown on other muck and upland soils, to see if this difference is a general one. Loss of Ascorbic Acid During Xtmage

Samples of spinach, cut as for market, were held at room tempera- ture for 17 days. Duplicate samples were kept at 1 to 3" C. (33.8 to 37.4' F.) . Leaves from each of t.he samples were analyzed for ascorbic acid (Table 4) at the start of the experiment and at the end of 3, 7, and 17 days. The spinach on standing for three days at room tempera- ture lost approximately one half of its ascorbic acid ; but after three days' storage at 1 to 3" C. (33.8 to 37.4" F.) the spinach retained practically all of its ascorbic acid. This shows the importance of refrigerating spinach soon after it has been cut.

SUMMARY AND COKCLUSIONS

The ascorbic acid contents of twelve varieties of spinach grown on upland soil were compared with that of the same varieties grown on

VITAMIN C CONTENT OF VEGETABLES

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muck soil, all samples being harvested and anaIyzed on the same day.

Those grown on the upland soil averaged 50 per cent higher in vitamin

C than those from the muck. While in both instances certain varieties ran somewhat higher than the average in ascorbic acid, yet the varietal differences are believed to be of secondary importance.

The vitamin C content of spinach as marketed is principally in the

leaves, the stems being nearly devoid of this vitamin.

Spinach stored at 1 to 3'

C. (33.8 to 37.4" F.) lost its ascorbic acid

very dowly, whereas that held at room temperature lost approximateIy

one haLf of its ascorbic acid in three days, and practically all of it in seven days.

TABLE 4

Loss of Ascorbic Acid from Spi~~h

during Storage

Variety of

spinach

Prickly Winter

Prickly Winter

Hollandia

Hollandia

Storage

in degrees C.

It0

3

23 to

26

lto

3

23 to

26

Ascorbic acid in leaves after storage for the

number of days indicated below

013171

17

mg.per gm

0.78

0.78

0.79

0.79

mg.pergm.

mg.gcrgm. mg. per gm.

0.76

0.72

0.39

0.44

0.76

0.05

0.64

Spoiled

0.49

0.39

0.03

Spoiled

The percentage of ascorbic acid in growing spinach leaves did not change materially during the harvesting period of one week.

REFERENCES

BESSEP,0. A., AND EING,

C. G., 1933.

The distribution of vitamin C in plant and

animal tissues, and its determination.

5. Biol. Chem. 103, 687 698.

EDDY,W. H., KOHblAN, E. F., AUD CAELsSON, v., 1925. Vitamins in canned foods.

111. Canned spinach. Ind. Eng. Chem. 17, 69-74.

  • v. HLHN,F . v., AND GORBING, J., 1933.

Inflnence of fertilizers on the vitanijn c

content of spinach. 2. Untersuch. Lebensm. 65, 601 16.

  • V. HAEIN,F . V., 1931.

Vitamin

studies. IV. The vitamin content of vegetables of

the German retail business. Z. Untersuch. Lebensm. 61, 545-610. HESSLER, M. C., AND CRAIG,G. C., 1929. Vitamin C content of spinach and Jona- than apples. Mo. Agr. Expt. Sta. Bull. 272, 67-8.

HESSLER,M.

C., JOEINSOH,

D., ANI, WrrZISH, R. B., 1931.

The vitamin content of

Bloomsdale spinach. Mo. Agr. Expt. Sta. Bull. 300, 83 84.

EIFER,

HILDA B ., AND MUNSELL,€1. E., 1932. Vitamin content of three varieties of

spinach.

3. Agr. Res. 44,

767-71.

SIIERNAN, H. C., AND SMITK,S. L., 1931. The Vitamins. 2nd Ed. ReinhoId Com- pany, New Pork City. TAUBEE,H., AND KLEINER,I., 1935. A method for the quantitative determination

of ascorbic acid (vitamin C).

J. Biol. Chem. 108, 563-70.

WASSON, G. E., 1931. Vitamin C content of fresh and canned spinaeh, Swiss chard, asparagus and carrots. 8. Dak. Agr. Expt. Sta. Bull. 261, 3-28.