A CONTRIBUTION TO THE CHEMISTRY OF TOMATO

PIGMENTS
THE COLORING MATTER IN AMERICAN RED AND PURPLE
TOMATOES (LYCOPERSICUM ESCULENTUM)*

BY M. B. MATLACK AND CHARLES E. SAND0

(From the Food Research Division, Bureau of Chemistry and Soils, United
States Department of Agriculture, Washington)

(Received for publication, November 28,1933)

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The characteristic red pigment of the tomato was first investi-
gated in 1876 by Millardet (l), who obtained it in a crystalline
state and also observed the crystals in the flesh of the ripe fruit.
He reported that the crystalline pigment, for which he proposed the
name solanorubine, was insoluble in water, soluble in hot alcohol,
and easily soluble in carbon disulfide, chloroform, and benzene.
He also established that absorption in the visible portion of the
spectrum of a carbon disulfide solution of the pigment was charac-
terized by two bands in the green region and one in the blue.
Later investigators (2-5) found carotene present in tomatoes
and considered it identical with the pigment obtained by Millardet.
Tomato pigment was again studied in 1903 by Schunck (6) who
found the red coloring matter clearly distinguishable from carotene
in appearance, crystal form, solubility, and absorption spectrum,
and named it lycopin. Since the work of Schunck the pigment
has usually been referred to as lycopin, although lycopersicin (7)
and lycopene have both been used. The latter term is to be pre-
ferred since the ending “ene” denotes the unsaturated hydrocarbon
nature of the coloring matter.
The first chemical investigation of lycopene was undertaken by
Montanari (8) in 1904. His results led him to consider the tomato
pigment a condensation product of 2 molecules of carotene, pos-
sessing the formula G2H74.

* Contribution No. 202 from the Food Research Division.

407
408 Chemistry of Tomato Pigments

In 1909 Willst%tter andEscher (9,lO) isolated from Italian tomato

conserve1 a relatively large quantity of the pure crystallized red pig-
ment, together with a much smaller quantity of the yellow pigment,
carotene. They found that lycopene is an unsaturated hydro-
carbon and possesses the same composition and molecular weight
as carotene, namely CNHS~. The crystals had a brownish carmine-
red color and velvety waxy consistency. Under the microscope
they generally appeared as needles or elongated, flat-sided prisms,
with split or blunt ends, and often occurred in clusters. Where
individual crystals crossed each other, there was observed a deeper

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red color. Lycopene was less soluble than carotene in ether,
carbon disulfide, petroleum ether, and alcohol, and oxidized and
bleached more readily.
As a result of the work of Karrer and others (11-16) the follow-
ing st’ructural formula has been assigned (14) to the pigment.

(3% CH,
I I
(CH,)&=CH-CHZ-CH,-(C-CH-CH=CH),--C=CH-CH-

CH, CHa
I
CH-CH=(C-CH=CH-CHL=C-CHz-CHF-CH=C(CH,)z

Willstatter and Escher, as well as Montanari, carried out their

investigations on the pigment isolated from Italian-grown varieties.
As far as we know, there has been no complete chemical investiga-
tion reported on the pigments occurring in tomatoes grown in the
United States. It is well known that the best known commercial
variaties of Italian tomatoes differ in many respects from domestic
varieties. Italian tomatoes are smooth, plum-shaped, and ap-
proximately 3 inches long and 1 inch in diameter. As a rule,
they have a more brilliant and more uniformly distributed red
color than the average tomato grown in the United States. They
have firm flesh and practically no core. American-grown toma-
toes are large and globular and generally more juicy. While
California tomatoes more closely resemble Italian-grown tomatoes
in composition, the latter are sweeter and contain less acid than
most domestic varieties. In this country two distinct types of red

1 Societk generale delle conserve aliment& cirio, Naples.

 M. B. Matlack and C. E. Sando 409
tomatoes are recognized. One type is carmine-red with a distinct
purplish cast. Tomatoes belonging to this type, which are charac-
terized by purplish red flesh and transparent skin, are sometimes
described as purple tomatoes. Typical examples are Livingston
Globe and Cooper Special. In the second type the color ap-
proaches scarlet-red. These tomatoes are characterized by
purplish red flesh and yellow pigmented skin, which together cause
bhe color to appear more nearly true red. The Stone, Indiana
Baltimore, and Santa Clara Canner are examples of this type.
Because of the difference mentioned above in appearance of

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American tomatoes and the more decided differences existing be-
tween American- and Italian-grown tomatoes, it was decided to
isolate and study the pigment from American red and purple
tomatoes and to determine whether it is identical with that isolated
from Italian varieties. The results of such an investigation would
definitely answer the question often raised by canners as to
whether the pigments are the same in American red and purple
varieties.
EXPERIMENTAL

Preparation of Crude Pigment-For the purpose of this study

four varieties were used, namely Fiaschetti (Italian-grown red),
Indiana Baltimore (American-grown red), Santa Clara Canner
(American-grown red), and Cooper Special (American-grown
purple). Commercially concentrated tomato pulp, either puree or
paste, was used in each case.
The method employed for the isolation of the crude pigment was
essentially that of Willstatter and Escher (10). When puree was
used the insoluble portion, which contained most of the pigment
associated with the plastids, was separated from the largest portion
of the juice by centrifuging in a De Lava1 cream separator, with a
clarifying bowl. Further dehydration was effected by the use of
95 per cent alcohol. After the mixture was stirred, the excess
aqueous alcoholic portion was removed by means of a hand-press
and the process repeated twice. When tomato paste was used,
it was necessary to carry out only the alcoholic dehydration
process.
The partly dried, dark red material which remained after treat-
ment with alcohol was completely dried in the open on a wire
 FIG. 1 FIG. 2

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FIG. 1. Rosettes of needles of lycopcne from carbon disulfide and pctro-
leum ether (X 28).
FIG. 2. Elongated, flat-sided prisms of lycopenc from carbon disulfide
and petroleum ether (X 90 approximat,ely).

FIG. 3. Spectrograms of lycopcne from Fiaschetti, A; Indiana Baltimore,

B; Cooper Special, C; Santa Clara Canner, D. The concentration is 1 mg.
per 100 cc. of an equal mixture of methyl alcohol and ethyl ether; 5 mm.
thickness; liquid air temperature; 1 minute exposure; tungsten light;
helium reference lines.
410
 M. B. Matlack and C. E. Sando 411

screen at a temperature not exceeding 40”. The product thus

obtained was finely ground and extracted with carbon disulfide in
a percolator until the extract was practically colorless. The
extract was then evaporated in vacua to small bulk at a tempera-
ture not exceeding 40”. To this concentrated pigment solution 3
volumes of absolute alcohol were added, and the mixture was kept
at low temperature until separation of the semicrystalline pigment
occurred. The insoluble pigment was collected by suction on a
porcelain funnel under a stream of nitrogen and when thoroughly
dried was stored in sealed tubes in the absence of oxygen. In all,

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57 gm. of the crude pigment were prepared in this manner.
PuriJication for AnalysisThe crude pigment was dissolved in
the smallest possible volume of carbon disulfide, and 2 to 3 volumes
of redistilled petroleum ether (b.p. 30-60”, known commercially as
Skellysolve) were added slowly with shaking. The crystalline
product which gradually separated was redissolved in carbon
disulfide and reprecipitated with petroleum ether several times.
While petroleum ether is superior to alcohol for the crystallization
of lycopene from carbon disulfide solution, there is a greater loss
due to greater solubility of the pigment in petroleum ether. Care
was taken at all stages to protect the pigment from oxidation by
keeping both the solution and filtered crystals under an atmos-
phere of nitrogen. As obtained by this method of purification and
crystallization, the crystals of lycopene exhibited a brownish
carmine-red color with a metallic sheen and appeared under the
microscope as rosettes of needles (Fig. 1) or elongated, flat-sided
prisms (Fig. 2).
The pigment from each of the four sources, after being dried in a
vacuum desiccator over phosphorus pentoxide, gave the following
analysis.
Fiaschetti
0.0685 gm.: 0.0651 gm. Hz0 and 0.0224 gm. COY
0.0676 “ : 0.0635 “ “ “ 0.2219 “ “
Found. C 89.18, 89.52; H 10.65, 10.53

Indiana Baltimore
0.0680 gm.: 0.0630 gm. Hz0 and 0.2222 gm. CO2
0.0680 “ : 0.0648 “ “ “ 0.2232 “ “
0.0764 “ : 0.0712 “ “ “ 0.2509 “ “
Found. C89.12. 89.52, 89.56; H 10.39, 10.68. 10.45
412 Chemistry of Tomato Pigments
Santa Clara Canner
0.0681 gm.: 0.0649 gm. Hz0 and 0.2234 gm. CO,
0.0633 “ : 0.0589 “ “ “ 0.2073 “ “
Found. C 89.47, 89.31; H 10.68, 10.43
Cooper Special
0.0710 gm.: 0.0654 gm. Hz0 and 0.2329 gm. CO,
0.0691 “ : 0.0642 “ “ “ 0.2275 “ “
Found. C 89.46, 89.79; H 10.33, 10.41
Found, average of nine determinations. C 89.44; H 10.51
Lycopene, G~HNT, requires: C 89.48; H 10.52

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Spectral Analysis
In order to obtain additional evidence of the identity and purity
of lycopene from the four sources, samples of each were submitted
to spectrographic analysis. The spectrograms shown in Fig. 3
were kindly prepared by G. E. Hilbert and E. F. Jansen of the
Bureau of Chemistry and Soils, United States Department of
Agriculture, by means of a new low temperature procedure de-
veloped by them. An examination of the absorption spectra
leaves no doubt that the coloring matters from all four sources are
identical.
Fig. 4 represents the visible and ultra-violet absorption spectrum
of lycopene which was dissolved in a mixture of ether and alcohol.
The data used in plotting this curve were kindly furnished by the
United States Bureau of Standards through the courtesy of K. S.
Gibson, D. B. Judd, and M. E. Brown. Since lycopene is easily
autoxidizable, it was found extremely difficult to prepare solutions
entirely free of oxidation. Even the same solution when allowed
to remain in the dark overnight showed evidence from the data
observed of some oxidation. Changes in the curve due to the
effect of slight oxidation were somewhat comparable to those
reported by MeNicholas (17) for carotene. The visible portion of
the curve represents a solution which was considered the best of
three samples. Owing to refinements introduced in its preparation
and to the nature of the curve obtained, it is believed that very
little if any oxidation could have taken place in this solution.
While the fullest precautions were exercised to prevent oxidation
of the solution used for the ultra-violet measurements, the curve
for this portion of the spectrum is probably not quite so accurate
as that for the visible end. Furthermore, while observations in
 M. B. Matlack and C. E. Sando 413
the ultra-violet region indicated an absorption band between the
frequencies 1000 and 1050, complete data were lacking. The
probable course of the absorption in this region is, therefore, indi-
cated by a dotted line.

WAVE-LENGTH ( Z’ip=MILLIMICRONS-METERS x 1O-g >

500 450 400 350 300 250

2.6

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2.4

= 1.6
is
a 1.4
3
m 1.2
<
1.0

0.6

0.6

0 A

0.2

600 100 600 900 1,000 1,100 1,200

FREQuENcY (VIBRATIONS t SECONDS x 10’~)

FIG. 4. Visible and ultra-violet absorption spectrum of lycopene. Sol-
vent: 80 per cent alcohol + 20 per cent ether; concentration: 0.40 cg. per
liter; thickness: 2 cm.; methods: clear circles, visual; solid circles,
photographic. Absorbancy is defined by the relation, - log10 (T.,L/T,,,.),
where the transmissions of the cells containing the solution and solvent
are represented by !/‘,,I, and T,,,., respectively; refer to McNicholas (17).

SUMMARY

The characteristic red coloring matter in American red and

purple tomatoes has been isolated and studied. The pigment
414 Chemistry of Tomato Pigments

obtained from the American-grown varieties, Indiana Baltimore,

Santa Clara Canner, and Cooper Special, was found to be identical
with lycopene isolated from an Italian variety known as Fiaschetti.

BIBLIOGRAPHY

1. Millardet, A., Note sur une substance colorante nouvelle (solano-

rubine) decouverte dans la tomate, Nancy (1876); abstracted in
Just’s Bot. Jahresber., 4, 783 (1876).
2. Arnaud, A., Compt. rend. Acad., 102, 1119 (1886).
3. Passerini, N., Staz. sper. agrar. ital., 18, 545 (1890).
4. Kohl, F. G., Untersuchungen uber das Carotin und seine physiologischen

Downloaded from http://www.jbc.org/ by guest on June 23, 2020

Bedeutung in der Pflanze, Leipsic, 41 (1902).
5. Ehring, C., Uber der Farbstoff der Tomate, ein Beitrag zur Kenntnis
des Carotins, Dissertation, Munich (1896).
6. Schunck, C. A., Proc. Roy. Sot. London, 72,165 (1903).
7. Duggar, B. M., Washington University Studies, 1, pt. 1, 22 (1913).
8. Montanari, C., Staz. sper. agrar. ital., 37,909 (1904).
9. Escher, H. H., Zur Kenntnis des Carotins und des Lycopins, Promo-
tionsarbeit, Zurich (1909).
10. Willstgtter, R., and Escher, H. H., Z. physiol. Chem., 64, 47 (1910).
11. Karrer, P., and Widmer, R., Helv. chim. acta, 11,751 (1928).
12. Karrer, P., and Bachmann, W. E., Helv. chim. acta, 12, 285 (1929).
13. Karrer, P., Helfenstein, A., and Wehrli, H., Helv. Aim. acta, 13,
(1930).
14. Karrer, P., Helfenstein, A., Wehrli, H., and Wettstein, A., Helv. chim.
acta, 13, 1084 (1930).
15. Karrer, P., Helfenstein, A., Pieper, B., and Wettstein, A., Helv. chim.
acta, 14, 435 (1931).
16. Kuhn, R., and Grundmann, C., Ber. them. Ges., 66,898 (1932).
17. McNicholas, H. J., Bur. Standards J. Research, 7, 185 (1931).
 A CONTRIBUTION TO THE
CHEMISTRY OF TOMATO PIGMENTS:
THE COLORING MATTER IN
AMERICAN RED AND PURPLE
TOMATOES (LYCOPERSICUM
ESCULENTUM)
M. B. Matlack and Charles E. Sando
J. Biol. Chem. 1934, 104:407-414.

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