Lebensm.-Wiss. U.-Technol.

36 (2003) 153–156

Research Note
Effect of thermal pasteurization on Valencia orange juice
color and pigments
Hyoung S. Lee*,1, Gary A. Coates
Florida Department of Citrus, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, USA
Received 11 February 2002; accepted 16 April 2002

Abstract

Changes in carotenoid pigment content and juice color due to thermal pasteurization of Valencia orange juices were studied. Total
carotenoid pigment content loss was significant (Po0.05) after thermal pasteurization at 90 C for 30 s. Thermal effects on
carotenoid pigment contents, especially on violaxanthin ( 46.4%) and antheraxanthin ( 24.8%), were clearly observed. With the
loss of violaxanthin and antheraxanthin, lutein became the major carotenoid, followed by zeaxanthin, in pasteurized Valencia
orange juice. There was perceptible color change after orange juice pasteurization, which led to juice color becoming lighter and
more saturated. Decreases in CIE a* value and increases in CIE L*, b*, h*, and C* are the major color changes after pasteurization.
Overall increases in reflected light might also influence the perception of color to a great extent in pasteurized orange juice. Total
color differences (DE*) compared to the fresh juice was 2.9270.97 (Po0.05).
r 2003 Published by Elsevier Science Ltd. on behalf of Swiss Society of Food Science and Technology.

Keywords: Pasteurization; Orange juice; Color; Carotenoid

1. Introduction enough for pigment loss to occur under the current

Pasteurization is important to the stability of citrus
juice during transportation and marketing. Carotenoids
are generally stable to heat treatment involved in the
common unit operations of food processing such as
blanching, cooking, and canning (Borenstein &
Bunnell, 1966), however, the stability of carotenoids in
foods varies greatly (Purcell, Walter, & Thompkins,
1969). Previously, it was assumed that neither pasteur-
ization nor concentration of orange juice had an
appreciable effect on carotenoid content (Higby, 1963).
However, a recent thermal processing study (Lessin,
Catigani, & Schwartz, 1997) with various fruits and
vegetables, including orange juice, indicated a re-
latively large loss (36%) of provitamin A carotenoids
(b-carotene, a-carotene and b-cryptoxanthin) in orange
juice with processing.
In red grapefruit juices, typical thermal pasteurization
(80–95 C for 15–30 s) did not appear to be severe
processing condition (Lee & Coates, 1999); there were
no significant losses in two major carotenoids, b-
carotene and lycopene, after thermal pasteurization.
However, carotenoid profiles in sweet oranges are more
complicated. In sweet oranges, oxygenated xantho-
phylls, representing more than 90% of the total
carotenoids (Philip, 1975), have a different thermal
stability compared to the hydrocarbon carotenoids such
as b-carotene and lycopene that are predominant in red
grapefruit.
Valencia orange is quantitatively the richest juice in
carotenoids, known to have the most complex pigment
pattern among sweet oranges (Gross, Gabai, & Lifshitz,
1972). Since Valencia orange is one of the principal
sweet orange varieties for processing, this study was
aimed to evaluate any pigment loss and potential visual
color changes associated with thermal pasteurization of
Valencia orange juice.

2. Materials and methods

*Corresponding author.
E-mail address: hsl1920@aol.com (H.S. Lee).
1
Lee is now associated with FDA, CFSAN, OFAS, College Park, Valencia orange juice was prepared using a household
MD 20740, USA. table-top citrus juicer (Waring, New Hartford, CT,

0023-6438/03/$30.00 r 2003 Published by Elsevier Science Ltd. on behalf of Swiss Society of Food Science and Technology.
doi:10.1016/S0023-6438(02)00087-7
154 H.S. Lee, G.A. Coates / Lebensm.-Wiss. U.-Technol. 36 (2003) 153–156
USA). Three sets of ten fruit each were weighed, hand-
squeezed, and filtered through cheesecloth. The juice
was then homogenized in an Omni mixer homogenizer
(Model 17105, Gainesville, VA, USA) at speed 4 for
1 min. Thermal pasteurization was performed by using
the GCA/Precision Thermo Circulating water bath
(Model 253, Chicago, IL, USA). The juice was heated
to 90 C for 30 s and followed by rapid chilling in an ice-
water-bath.
2.1. Color analysis
Color (CIE L*, a*, b*) analysis was conducted with
a Macbeth COLOR-EYEs 3100 spectrophotometer
(Kollmorgen Instruments Corp., Newburgh, NY, USA)
with Optiviews software package in the reflectance
mode as previously described by Lee and Castle (2001).
Also, CIE L*, a*, b* values are used to calculate the hue
angle (tan 1 b*/a*), chroma {(a*2+b*2)1/2}, and total
color differences {(DL*2+Da*2+Db*2)1/2} before and
after pasteurization (Lee & Coates, 1999).
2.2. Analysis of carotenoids
Carotenoid pigments were extracted, saponified, and
analysed by RP-HPLC method according to a proce-
dure described by Lee (2001). Also, estimation of total
carotenoid content was carried out as described by Lee
(2001).
2.3. Statistical analysis
All experiments were conducted on triplicate samples.
All statistical analyses were conducted using the
SigmaStat PC software from SPSS, Inc. (Chicago, IL,
USA). Results were submitted to analysis of variance
(ANOVA), regression analysis, and least significant
differences (LSD) with significance defined as Po0.05.
3. Results and discussions
3.1. Effects on pigment contents
The major carotenoid pigments, which are responsible
for visual color of the fresh and pasteurized juices,
were measured by HPLC (Fig. 1) and are presented in
Table 1. Carotenoids are identified based on our
previous work with sweet orange juices (Lee, 2001)
under the same HPLC condition. In the fresh Valencia
orange juice, cis-violaxanthin and antheraxanthin are
the two primary carotenoids as previously presented by
Curl and Bailey (1959).
Total pigment content, expressed as the sum of
carotenoid concentrations measured at 450 nm, was
6.2570.11 mg/L for fresh juices. In the past-
eurized juices, total pigment content decreased to
5.7070.08 mg/L, which is about a 10% loss (Po0.05).
Among 22 carotenoid peaks presented in Table 1, nine
of them showed significant changes (Po0.05) under the
same thermal processing condition at 90 C for 30 s.
Most pigment loss was from 5,6-epoxide carotenoids
such as violaxanthin ( 46.4%), cis-violaxanthin
( 19.7%), and antheraxanthin ( 24.8%). With the loss
of major 5,6-epoxide carotenoids during thermal pas-
teurization, the carotenoid pattern in Valencia orange
juice changed. Lutein became the primary carotenoid
followed by zeaxanthin (Table 1).
It was also interesting to observe the slight increases
in 5,8-epoxide carotenoids, luteoxanthin and mutato-
xanthin, after thermal pasteurization. Disappearance of
5,6-epoxide, with accompanying increases of 5,8-epox-
ides in greater proportions, was previously observed
during the preparation and storage of powdered
Valencia orange juice (Curl & Bailey, 1959). Viola-
xanthin is one of the most labile carotenoids and is
easily isomerized in the presence of acid to luteoxanthin
and then to auroxanthin (Rodriguez-Amaya, 2000). The
disappearance of violaxanthin, the principal carotenoid
of the fresh mango, was often reported in processed
Mango juices (Mercadante & Rodriguez-Amaya, 1998;
Rodriguez-Amaya, 2000).
However, changes in carotenes with provitamin A
activity (b-carotene, a-carotene and b-cryptoxanthin)
are relatively small compared to the xanthophylls; thus,
no significant (P>0.05) losses in provitamin A activity
were observed after thermal pasteurization.
3.2. Effects on color parameters
After thermal treatment, CIE b* values (17.6270.35)
gradually changed toward a more positive direction
(20.0271.05) in all samples, but CIE a* values shifted to
a slightly negative direction from 1.7570.07 to
2.6470.15 (Po0.05). It was a different direction in
color change compared to the color changes observed in
red grapefruit juice (Lee & Coates, 1999). Thus, a color
shift toward positive b* and negative a* directions
indicate more yellow and less red in pasteurized juices.
The majority of samples showed slight increases in CIE
L* value after pasteurization, which indicates a light-
ening of juice surface color. A small increase in CIE L*
value from 40.2270.16 to 41.2270.81 for pasteurized
juices can probably be attributed to partial precipitation
of unstable, suspended particles in juice as described by
Genovese, Elustondo, and Lozano (1997). A similar
observation of small increases in CIE L* values was also
reported with thermal processing of red grapefruit juices
by Lee and Coates (1999).
Total color differences (DE*), which indicate the
magnitude of the color difference between fresh and
pasteurized juices, are 2.9270.98 (Po0.05). It has been
 H.S. Lee, G.A. Coates / Lebensm.-Wiss. U.-Technol. 36 (2003) 153–156
Fig. 1. Changes in carotenoid profiles in Valencia orange juice before (A) and after (B) thermal pasteurization at 90 C for 30 s. See Table 1 for peak
identification
considered (Francis & Clydesdale, 1975) that a DE* of 2
would be a noticeable visual difference for a number of
situations.
The mean value for the hue (h*) angle of fresh juices is
95.66. The hue angle for pasteurized juices was slightly
(Po0.05) shifted to 97.51, probably due to changes in
pigment profile. Chroma (C*), which represents color
intensity, increased from 17.70 to 20.19 (Po0.05) after
pasteurization. A similar observation in increases in
chroma (increases in brightness) due to the application
of heat for the manufacture of tomato juice has been
previously described (Davis & Gould, 1955).
There are also differences in the magnitude of
reflected light in juices between before and after
pasteurization as measured by spectral reflectance
graphs (not presented). Reflectance values (%) deter-
mined from the 350 to 700 nm range were compared
before and after pasteurization. After pasteurization, the
reflectance spectrum changed significantly (Po0.05); the
amount of reflectance light in the pasteurized juice
increased. There were more changes throughout the
155
visible region of the spectra, especially above 550 nm,
which means that relatively more yellow–red light will
be reflected to the eye and that the samples will appear
to increase in the yellow–red region. In view of the
overall increase in reflected light due to pasteurization, a
slightly lighter and more vivid juice color can be
expected, as observed with red grapefruit juice (Lee &
Coates, 1999).
4. Conclusions
Thermal effects on losses of carotenoid contents,
especially on violaxanthin, antheraxanthin, and cis-
violaxanthin, were clearly detected. Isomerizations of
the main 5,6-epoxide carotenoids to 5,8-epoxides are
probably responsible for the color changes after thermal
juice pasteurization. There was perceptible color change
during orange juice pasteurization, which led to juice
color becoming lighter and more saturated.
156 H.S. Lee, G.A. Coates / Lebensm.-Wiss. U.-Technol. 36 (2003) 153–156

Table 1
Carotenoid compositions in fresh and pasteurized Valencia orange juice

Peak no. tR (min) Fresh (mg/L)a Pasteurized (mg/L)a lmax (nm) Identification

1 5.27 0.108 0.083** 418,441,470 Neoxanthin a

2 5.69 0.023 0.019 419,441,470 Neoxanthin b
3 6.05 0.019 0.016 401,423,449 Neochrome
4 7.54 0.046 0.040 324,414,438,466
5 7.86 0.186 0.171 329,419,446,472
6 8.00 0.148 0.139 419,440,467
7 8.73 0.574 0.308** 414,442,472 Violaxanthin
8 9.81 0.070 0.057** 332,438,467
9 10.36 0.387 0.351** 400,424,450 Luteoxanthin
10 11.51 0.810 0.655** 328,414,438,466 cis-Violaxanthin
11 12.06 0.819 0.616** 422,447,475 Antheraxanthin
12 12.75 0.162 0.212** 397,419,445 Luteoxanthin
13 13.76 0.132 0.230** 404,423,455 Mutatoxanthin
14 14.33 0.670 0.760** 426,443,474 Lutein
15 15.50 0.582 0.534 333,420,444,471 Isolutein
16 16.18 0.662 0.662 428,452,481 Zeaxanthin
17 18.22 0.044 0.051 342,449,476
18 19.61 0.128 0.129 425,447,477 a-Cryptoxanthin
19 22.13 0.337 0.326 429,452,479 b-Cryptoxanthin
20 24.80 0.013 0.016 379,400,424 z-Carotene
21 26.19 0.084 0.087 419,449,476 a-Carotene
22 28.84 0.130 0.129 429,454,479 b-Carotene
a
Mean of triplicate samples; based on calculation from total carotenoids at 450 nm.
tR (min) is retention time and lmax (nm) is absorption maxima.
Means followed by asterisks indicate significant difference (Po0.05).

References Lee, H. S., & Castle, W. S. (2001). Seasonal changes of caro-

Borenstein, B., & Bunnell, R.H. (1966). Cartenoids, properties,
occurrence, and utilization in foods. In C.O. Chichester, E.M.
Mrak, G. F. Stewart (Eds.), Advances in food research, (Vol. 15,
pp. 175–195). New York: Academic Press.
Curl, A. L., & Bailey, G. F. (1959). Changes in the carotenoid
pigments in preparation and storage of Valencia orange juice
powder. Food Technology, 13, 394–398.
Davis, R. B., & Gould, W. A. (1955). The effect of processing methods
on the color of tomato juice. Food Technology, 9, 540–547.
Francis, F. J., & Clydesdale, F. M. (1975). Food colorimetry: Theory
and applications. Westport, CT: The AVI Publishing Co., Inc.
Genovese, D. B., Elustondo, M. P., & Lozano, J. E. (1997). Color and
cloud stabilization in cloudy apple juice by steam heating during
crushing. Journal of Food Science, 62, 1171–1175.
Gross, J., Gabai, M., & Lifshitz, A. (1972). A comparative study of the
carotenoid pigments in juice of Shamouti, Valencia, and Washington
oranges, three varieties of Citrus sinensis. Phytochemistry, 11, 303–308.
Higby, W. K. (1963). Analysis of orange juice for total carotenoids,
carotenes, and added beta-carotene. Food Technology, 17, 95–98.
Lee, H. S. (2001). Characterization of carotenoids in juice of red Navel
orange (Cara Cara). Journal of Agricultural and Food Chemistry,
49, 2563–2568.
tenoid pigments and color in Hamlin, Earlygold, and Budd
Blood orange juices. Journal of Agricultural and Food Chemistry,
49, 877–882.
Lee, H. S., & Coates, G. A. (1999). Thermal pasteurization
effects on color of red grapefruit juices. Journal of Food Science,
64, 663–666.
Lessin, W. J., Catigani, G. L., & Schwartz, S. J. (1997). Quantification
of cis–trans isomers of provitamin A carotenoids in fresh and
processed fruits and vegetables. Journal of Agricultural and Food
Chemistry, 45, 3728–3732.
Mercadante, A. Z., & Rodriguez-Amaya, D. B. (1998). Effects of
ripening, cultivar differences, and processing on the carotenoid
composition of Mango. Journal of Agricultural and Food Chem-
istry, 46, 128–130.
Philip, T. (1975). Carotenoid esters in plant products. Food Technol-
ogy, 29, 50,54.
Purcell, A. E., Walter Jr, W. M., & Thompkins, W. T. (1969).
Relationship of vegetable color to physical state of the carotenes.
Journal of Agricultural and Food Chemistry, 17, 41–42.
Rodriguez-Amaya, D. B. (2000). Factors influencing carotenoid
composition in foods. In The fourth international symposium on
natural colorants (pp. 258–263). Hamden, CT: The Hereld
Organization.