VITAMIN C CONTENT O F VEGETABLES. I.

SPINACH”

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

N e w York State Agricultural E x p e r i m e n t Station, Geneva, N e w Pork, alzd the
Department of Chemistry, University of Pittsburgh, Pittsbwgh, PennsyEva.nia

(Received for publication, October 23, 1935)

Fresh spinach has long been known t o 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 f o r protection of a guinea
pig from scurvy, the data in recent papers may be recalculated on a
uniform basis (Table I.).
TABLE 1
Previous R e p o r t s OR the Asambic Acid Content of Spinach

Investigator Source or rariety Ascorbic acid

of spinach
mg. p e r 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 t o 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 t o 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.
3

FOOD
RESEARCH,FOL.1, No. 1
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 t o 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 t o 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 t o permit them t o reach approxi-
mately 325 gm. in weight. They were then given the basal diet only f o r 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 t o each animal,
The quantity to be fed was calculated from the indophenol titration value. T h e
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 v h m i n solution, fed from graduated pipettes.
I t is evident from the data (Table 2) t h a t the animal assays correspond closely
with the indophenol titratiou values.

EXPERIMENTAL
Relatz/ue Potency of Leaf a i d 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 O F VEGETABLES 5

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 i n euam-
ining spinach, either the entire plants should be taken o r else the
TABLE P
Biological Assay of Yitamin G in S@ncbeh LFfiWe8

Scu rr?
Basal diet supplement No. of Vitamin C Initial Gain in weight scare at
animals fed per day weight during 14 days autopsy
w. gni . gm .
Prickly Winter Spinach 4 0.5 320 38 1
4 1 .o 319 17 0
Holitlndia Spinach 4 0.5 307 35 2
4 1.0 309 43 0
Ascorbic Acid Solution 7 0.5 311 41 2
6 1.0 310 63 0
llnsupplemented 6 0.0 329 -113 19

leaires and stems should be assayed separately; otherwise it would be

difficult t o obtain uniformly reliable data. Since in spinach the ascor-
bic acid is principally in the leaves, the data presented in this paper
refer t o 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). T o 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 i t had nearIy attained full
growth, and again when it began to bolt. On these dates t h e 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 d a y loam a t Geneva, N. Y . ) and the same
varieties harvested on the same day from muck soiI a t Holcomb, N. Y.
One sample from each lot was analyzed f o r 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) t o 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 N w &and Upland Soil aad Harvested on the Same Day

Ascorbic acid in leaves harvested

from
Variety of spinach
Mucksoil I Uplandsoil
om.
mg. p a r m g . 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 o f 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 a t 1t o 3" C. (33.8 to
37.4' F.) . Leaves from each of t.he samples were analyzed for ascorbic
acid (Table 4 ) a t the start of t h e 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 a t 1 t o 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 7

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 a t 1t o 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 S p i ~ during

~ h Storage

Storage
Ascorbic acid in leaves after storage for the
Variety of number of days indicated below
spinach in degrees C.
0 1 3 1 7 1 17
mg.per gm mg.pergm. m g . g c r g m . mg. p e r gm.
Prickly Winter It0 3 0.78 0.76 0.72 0.39
Prickly Winter 23 t o 26 0.78 0.44 0.05 Spoiled
Hollandia lto 3 0.79 0.76 0.64 0.49
Hollandia 23 t o 26 0.79 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.

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