A Research Note
THERMOSTABILITY OF RED BEET PIGMENTS (BETANINE AND
VULGAXANTHIN-I): INFliUENCE’ OF ,pH AND TEMPERATURE
I. SAGU Y
-ABSTRACT
The thermostability of red beet pigments (betanine and vulgaxan-
thin-I) was investigatedin beet juice under atmosphericconditions
at 61.5, 75.5 and 85.5”C and at pH range4.8-6.2. The degradation
of both pigments followed first-order reaction kinetics. The energies
of activation were 19.2 * 0.5 and 16.3 f 0.6 Kcal/mole for betanine
and vulgaxanthin-I respectively,and were independent of pH. Max-
imum thermostability was observedat pH 5.8 for both pigments. In
beet juice under atmospheric conditions vulgaxanthin-I was more
sensitivethan betanine.
INTRODUCTION
AS A CONSEQUENCE of intensified safety awareness and
restrictions imposed on synthetic color compounds for use
in foods, considerable attention has been focused in recent
years on natural colorants. Only recently (Anon., 1978),
the ‘National Cancer Institute released a report which con-
cluded that p-cresidine, basic to the manufacture of FD&C
Red No. 40, one of the two (FD&C red No. 40 and No. 3)
remaining red food colors on the GRAS list, is carcinogenic
in the rats and mice in the NIC’s bioassay program.
Red and golden cultivars of beet root appear to be one
of the most logical sources of water-soluble colorants. The
color of beet consists of two major pigments betanine (red)
and vulgaxanthin-I (yellow).
The effect of pH on the stability of these pigments has
been studied in a variety of conditions and processes, such
as: sterilization (Habib and Brown, 1956; Lusas et al.,
1960); food products (von Elbe et al., 1974a); solutions
(Kopelman and Saguy, 1977; Peterson and Joslyn, 1960;
Sapers and Horenstein, 1978; Saguy et al., 1978a; Savolai-
nen and Kuusi, 1978; von Elbe et al., 1974b); extraction in
air or nitrogen (Wiley and Lee, 1978; Wiley et al., 1979).
However, the reported data are in some cases difficult to
evaluate because factors such as enzymatic activity, dilution
effect, oxygen concentration, pH, cultivar, and light were not
always considered.
This paper presents data on pH and temperature effect
on betanine and vulgaxanthin-I thermostability in beet
juice.
EXPERIMENTAL
BEET JUICE was obtained by homogenizingblanched beet slicesin
a Waring Blendor with an equal weight of McIlvaines O.lM buffer
solution of various pH ranging from 4.8-6.2 at room temperature.
The slices were blanched in steam for 4 min to prevent enzymatic
activity. The juice was filtered (Whatman No. 1) and 20 ml were
filled into vials. The unsealedvials were placed in a thermostatically
controlled (+ O.l”C) shaking water bath, for the heat treatment
exposure. The vials (triplicates) were withdrawn periodically and
Author Saguy, formerly affiliated with Technion, the Israeli Insti-
tute of Technology, is now with the Department of Nutrition
Food Science, Room 56-107, Massachusetts institute of Techno-
logy, Cambridge, MA 02 139.
0022-1147/79/0005-1554$02.25/O
01979 Institute of Food Technologists
1554-JOURNAL OF FOOD SCIENCE-Volume 44 (1979)
cooled down immediately in an ice water bath. The sampleswere
furthei diluted with the appropriate pH buffer to allow direct ab-
sorbarice measurementi within the range 0.2-0.8 optical density.
Color lcontent was analyzed according to the method described by
Saguyjetal. (1978b).
Regressioncoefficient determined on the kinetic data describing
the pigment losseswere obtained by BMD 02R (Dixon, 1971).
RESULTS & DISCUSSION
:I
THE CONCENTRATION of the retained betanine and vul-
gaxanthin-I monitored in the beet juice under atmospheric
$condi&ons exposed to various heat treatments and time,
,gener&ed a straight line when plotted on a semilogarithmic
$.cale,“for each pH tested, indicating a first-order reaction
lfor bgth pigments studied. The first order kinetics are in
Iiagreetient with previously reported data (von Elbe et al.,
11974g; Sapers and Hornstein, 1978; Savolainen and Kuusi,
~1978;/ Saguy et al., 1978a). The first-order reaction kinetics
permitted reaction rates to be expressed alternatively in
‘terms;of half-life values, T1 2.
Tefnperature and, pH e4fects on betanine and vulgaxan-
thin-1 thermostability are illustrated in Tables 1 and 2 re-
spectively. Maximum stability was observed at pH 5.8 for
,both pigments. Previous estimates of optimal pH were re-
ported as “between 5 and 6” (von Elbe et al., 1974b; Savo-
lainen’: and Kuusi, 1978).
Tehperature effect on pigments lability is expressed by
the energy of activation or alternatively by the “so called”
“Q1 0 value. Energies, of activation showed different values
for the two pigments, (Tables 1 and 2), e.g. 19.2 rf: 0.5 and
~16.3 + 0.6 Kcal/mole for betanine and vulgaxanthin-I, re-
spectively. The pH effect within the range 4.8-6.2 was
negligible. The energy of activation for betanine is substan-
tially ‘higher than reported by von Elbe et al. (1974b) (at
,pH S), e.g. 12.5 + 2.0 and 10.0 f 2.0 Kcal/mole in model
system and beet juiCe, respectively. Sapers and Hornestein
(1978) also reported betanine degradation rate constant at
2S°C ‘and pH 5 which was significantly lower than reported
by vdn Elbe et al. (1974b) (e.g. 0.066 vs 0.88 day-‘). A
possible reason for the discrepancies is betanine decolora-
tion &e to oxidation, which is particularly important at
relati$ely low temperatures and long times of exposure. Un-
der these conditions differences in dissolved oxygen con-
centration can furnish an explanation of the differences in
observed rates. Moreover, the excellent agreement found
when’comparing the half-life time obtained at 75.5’C with
values reported recently by Pasch and von Elbe (1979) sup-
ports,ithe assumption, that discrepancies exist mainly at low
temperatures and are due to differences in oxidative decol-
~oratiqn.
Unblrinched beet juice under atmospheric conditions and
modeiate temperatties is very susceptible to enzymatic de-
coloration. The activity (polyphenol oxidase) was the topic
& of nukerous studies (Lee and Smith, 1978; Soboleva et al.,
,,1976; Viner, 1977). Results indicated a maximum enzyme
potency in the range 25-43OC; however, residual activity
was noted even after a relatively high temperature long time
expodure (Lee and Smith, 1978; Wiley et al., 1979). Betacy-
anines are more susceptible to this enzymatic decoloration
than betaxanthines (Kopelman and Saguy, 1977; Wiley et
 THERMOSTABILITY OF RED BEET PIGMENTS. ..

Table l-Effect of temperature and pH on betanine thermostability in beet juice (kb first order rate constant, min-‘; T, i2 half life, min)

pH 4.8 pH 5.2 pH 5.8 pH 6.2

Temperature
(“C) kbx103 T, 12 kbx103 T, 11 kbx103 T, /z kbXl@ T, 12

61.5 6.3 110.1 5.6 124.5 4.5 154.3 5.5 126.6

75.5 24.3 28.5 16.5 41.9 14.6 47.4 16.8 41.2
85.5 40.5 17.1 37.4 18.5 32.0 21.7 40.5 17.1
1oo.oa 113.0 6.1 98.0 7.1 94.6 7.3 117.7 5.9

Energy of activation
(Kcal/mole) 18.8 18.8 19.6 19.7
0, ,, (70-80°C) 2.19 2.19 2.27 2.27

a Calculated

Table P-Effect of temperature and pH on vulgaxanthin-I thermostability in beet juice (k,-first order rate constant, min.‘; T, ,z half-
life mini
PH 4.8 pH 5.2 pH 5.8 pH 6.2
Temperature
(“C) k,x 10’ T,lz k,,xlO’ T&l, k,,xlO’ T,/* k,x103 T,/,
61.5 11.9 58.2 9.5 72.6 8.8 78.6 9.1 76.5
75.5 34.1 20.4 25.1 27.8 23.4 29.7 24.3 28.5
85.5 56.0 12.4 49.7 13.6 45.6 15.4 49.3 14.0
1 oo.o= 133.7 5.2 120.4 5.8 114.6 6.1 123.9 5.6
Energy of activation
(Kcal/mole) 15.5 16.3 16.5 16.8
0, o (70-80°C) 1.91 1.97 1.99 2.01

a Calculated

Lee. C.Y. and Smith. N.L. 1978. Polyphenol oxidase activity in

al., 1979) while the opposite behavior was observed in table beets. Presented at the 38th Annual Meeting of the Insti-
blanched juice (Tables 1 and 2). tute of Food Technologists, Dallas, TX.
The energy of activation values for vulgaxanthin-I were Lusas. E.W., Rice, A.C. and Weckel, K.G. 1960. Changes in the color
of canning beets. Res. Bull. 218. University of Wisconsin, Mad-
slightly affected by the beet juice pH (Table 2), however, ison, WI.
the difference observed was not significant. The calculated Pasch, J.H. and van Elbe, J.H. 1979. Betanine stability in buffered
solutions containing organic acids, metal cations, antioxidants, or
values at 1OO’C of vulgaxanthin-I degradation rate con- sequestrants. J. Food Sci. 44: 72.
stant, k,, and the half-life, T1 /2, are in general agreement Peterson, R.G. and Joslyn, M.A. 1960. The red pigment of the root
with previously reported values for purified pigment solu- of the beet (Beta
429.
vu&z&s)
-
as a pyrrole compound. Food Res. 25:

tions (Savolainen and Kuusi, 1978) but differ substantially Saguy, I., Kopelman, I.J. and Mizrahi, S. 1978a. Thermal kinetic
from those reported for beet juice. This discrepancy may be deaadation of betanine and betalanic acid. J. Agri. Food Chem.
2612): 360.
to some extent attributed to different atmospheric condi- &guy, I.,Kopelman, I.J. and Mizrahi, S. 197813. Computer aided
tions (air vs nitrogen). determination of beet pigments. J. Food Sci. 43: 124.
Sapers, G.M. and Hornstein. J.S. 1978. Varietal differences in colo-
It might be worth emphasizing that substantial thermo- rant properties and stability of red beet pigments. Presented at
stability differences were found in beet pigments as af- the 38th Annual Meeting of the Institute of Food Technologists,
fected by different systems (solution, beet juice and slices), Dallas,
Savolainen.
TX.
K. land Kuusi. T. 1978. The stability properties of gol-
water activity, atmosphere (oxygen, nitrogen), pH, light, den beet and red beet pigments: influence of pH. temperature
beet variety, organic acids, metals and other factors. Thus, and some stabilizers. Z. Lebensm. Unters.-Forsch 166: 19.
Soholeva, G.A.. Ul’yanova, M.S., Zakharova, N.S. and Bokuchava,
special attention is required when data are compared or M.A. 1976. Betacynine-decolorizing enzyme. Biokhimiya. 41:
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Viner, Y. 1977. Enzymatic degradation of red beet pigments (beta-
nish some explanation to the forementioned phenomena. nine and vulgaxanthin-I). B.Sc. thesis (Hebrew), Technion-Israel
Institute of Technology.
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Lindsay, R.C. 1974a. Evaluation of betalaine pigments as sausage
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Volume 44 (1979)-JOURNAL OF FOOD SCIENCE- 1555