Eur Food Res Technol (2006) 223: 282–289
DOI 10.1007/s00217-005-0202-z
ORIGINAL PAPER
Hou-yin Wang · Xiao-song Hu · Fang Chen ·
Ji-hong Wu · Zheng-hua Zhang · Xiao-jun Liao ·
Zheng-fu Wang
Kinetic analysis of non-enzymatic browning in carrot juice
concentrate during storage
Received: 14 September 2005 / Revised: 13 November 2005 / Accepted: 21 November 2005 / Published online: 28 December 2005


C Springer-Verlag 2005
Abstract The effect of storage temperature, storage time
and total soluble solid (TSS) on the total sugar (TS), su-
crose, glucose, fructose, total amino acid (TAA), pH, 5-
hydroxymethylfurfural (HMF) and browning degree (BD)
in carrot juice concentrate (CJC) stored at −18, 0, 25 and
37 ◦ C was studied and the relationship between HMF and
BD was established. Higher temperature at 25 and 37 ◦ C
had a significant effect (p<0.05) on TS, sucrose, glucose,
fructose, TAA, pH, HMF and BD, while lower temperature
at 0 and −18 ◦ C had less effect, and the storage time and the
soluble solid of CJC stored at 25 and 37 ◦ C also had a sig-
nificant effect on HMF formation and BD. At 25 and 37 ◦ C,
the formation of HMF and BD followed a first-order reac-
tion well as a function of storage time; a good correlation
between HMF formation and BD occurred. The formation
of HMF could described by the total soluble solid value
[TSS] and the BD value ([BD], absorbance at 420 nm) of
CJC, the equation was [HMF] (content of HMF) = 1.2177
− 0.1124 × [TSS] + (2.835 + 0.3477 × [TSS]) × [BD]
and [HMF] = 1.5510 − 0.4813 × [TSS] + (23.8847 +
0.7249 × [TSS]) × [BD] at 25 and 37 ◦ C, respectively.
Keywords Carrot juice concentrate (CJC) .
5-Hydroxymethylfurfural (HMF) . Browning degree
(BD) . Total soluble solid . The first-order reaction
Introduction
The carrot is an important root vegetable and is often used
for juice production [1]. Carrot juice has a high nutritional
value, as it is an important source of carotene [2]. Carrot
juices are preferably used as a natural source of provi-
H.-y. Wang · X.-s. Hu · F. Chen · J.-h. Wu · Z.-h. Zhang ·
X.-j. Liao (
) · Z.-f. Wang
College of Food Science & Nutritional Engineering, China
Agricultural University,
Beijing 100083, China
e-mail: liaoxjun@hotmail.com
Tel.: +8610-62737434
Fax: +8610-62737434
tamin A in the carotenoids drinks [3, 4]. Of the various
carotenoids in carrots, beta-carotene constitutes a large
portion, followed by alpha-carotene and lutein [5–7]. The
needing of consuming carrot juice concentrate (CJC) is
increasing very fast in China.
It is known that many food products darken during ther-
mal processing and storage. During storage of fruit and veg-
etable juice concentrate, the darkening is attributed mainly
to non-enzymatic reactions in apple juice [8–10], citrus
juice [11], peach juice [12], and in mei liqueur [13]. The
non-enzymatic reactions involve Maillard reaction, ascor-
bic acid degradation and caramelization. Caramelization
occurs on heat treatment of sugars at high temperatures;
ascorbic acid degradation occurs by an oxidative path in
citrus juices [11]. Maillard reaction, taking place between
alpha-amino groups and reducing sugars, may be desirable
during food processing as in the manufacture of coffee,
tea, beer and in the toasting and baking of bread. This re-
action improves desirable sensory characteristics of these
foods, e.g. color, aroma, and flavor [14–16]. However, it
may be undesirable in concentrated, intermediate mois-
ture, and dried foods [14]. In addition, Maillard reaction
causes losses in nutritional value of foods [17–19].
Previous studies reported a method of producing the car-
rot juice [20, 21], the stability of carrot juice [22–24], and
the nutrient loss during the processing or storage [25–27].
During the storage, CJC was susceptible to non-enzymatic
browning, which could impart the color to carrot juice.
However, few reports dealt with non-enzymatic browning
in CJC.
The objectives of this work was to analyze the kinetics
of non-enzymatic browning in CJC and to establish the
relationship between 5-hydroxymethylfurfural (HMF) for-
mation and browning degree (BD).
Materials and methods
Preparation and storage of CJC
The carrot defined as No. 1 Orange-red variety grown
in Beijing Vegetable Experimental Station of the Chinese

Carrots
Washing
cut off petiole hand peel
Slicing and
Citric acid addition
blanching 0.2%
90 ˚C, 3 min
Coarse grinding
grinding mill
Pectinex Smash
XXL 100ml/T
Enzymatic treatment
45˚C, 1 h
Enzyme inactivation
90 ºC,3 min
Pressing and
Pre-filtration
Citric acid addition
Filtration adjusted to pH 3.77
200 mesh
Rotary evaporator
60 ~ 65 ˚C, 110 mbar
Pasteurization
95 ˚C, 10min
Fig. 1 Flow chart of carrot juice concentrate (CJC) production
Academy of Agriculture Science in 2004 autumn season.
After harvesting, the fresh carrot was transported to the
laboratory by truck at about 20 ◦ C, and stored in woven
polypropylene bags (75 g m−2 ) in batches of approximately
15 kg each. Storage temperature was 0±1 ◦ C and the rela-
tive humidity was 85–95%. The storage time was less than
10 days before processing.
Two hundred kilograms of carrots in the study were used.
CJC was processed using the flow chart shown in Fig. 1.
Fresh carrot was washed with tap water, and blemished
carrots were discarded. Then carrots of good quality were
cut into slices about 2 cm thick. In experiments, the carrot
slices were acid blanched with citric acid (0.2%) at 90 ◦ C
for 3 min, which was proven to be sufficient for complete
pectinesterase inactivation, and pulped by a mill (JM-80,
Langfang Tongyong Machinery Co. Ltd, China). Prior to
enzymatic maceration, the pH of the pulp was adjusted
to about 4.8. The enzymatic maceration was performed
with 100 ml T−1 Pectinex Smash xxl (Novozymes Co.,
Beijing, China) for 1 h at 45 ◦ C. Pectinex Smash xxl was
inactivated by heating to 90 ◦ C for 3 min, and the pulp
was cooled to room temperature. The pressing of juice was
finished with a juicer, four-folded cheesecloth was firstly
used for pre-filtration of carrot juice, and then the juice was
filtered through 200-mesh filter sheet. The resulting juice
was added with potassium sorbate (1 g kg−1 ); its pH value
was readjusted to 3.77 with citric acid, and concentrated
with a rotary evaporator at 60–65 ◦ C and 110 mbar. The
same CJC was diluted to 20 , 40 and 60 ◦ Brix, respectively,
283
using de-ionized water (2000D De-ionized water machine,
Changfeng Instrument Co. Ltd., Chian).
All the samples of CJC with 20, 40, and 60 ◦ Brix were
filled into 150 ml glass bottle, pasteurized at 95 ◦ C for
10 min, and then stored in dark at −18, 0, 25 and 37 ◦ C
for 150 days. HMF formation and BD was measured every
30 days. Citric acid was purchased from Beijing Chemicals
Co. (Beijing, China) of analytical grade.
Preparation of samples
The CJC of 20, 40, and 60 ◦ Brix was diluted to 8.0
◦
Brix samples with de-ionized water, respectively. Some
8.0 ◦ Brix samples were directly used for the determination
of L-ascorbic acid, pH, and total amino acid (TAA). Some
8.0 ◦ Brix samples were centrifuged at 10,000×g at 4 ◦ C for
30 min. The supernatant was passed through a 0.45 µm cel-
lulose nitrate membrane filter (Shanghai Yadong Nuclear
Grade Resins Co. Ltd., China). The filtrates were separated
into three parts: one for BD measurement, second for HMF
analysis, and the third for sugar analysis.
Determination of 5-hydroxymethylfurfural (HMF)
Chromatographic determination of HMF was carried out
by modifying the method of Rada-Mendoza, et al. [28],
the high-performance liquid chromatography (HPLC) sys-
tem was composed of a K-1001 pump (Knauer Co., Ger-
many), a reversed C18 column (4.6×150 mm i.d.; Kromosil,
Switzerland), a K-2501 UV-detector (Knauer, Germany) set
at 283 nm, and a reversed C18 guard column (4.6×15 mm;
Kromosil, Switzerland). The mobile phase consisted of
methanol:water (10:90, v:v), the flow rate was 1 ml min−1
at 30 ◦ C, and the injection volume was 20 µl. Quantifica-
tion was carried out by the external standard method using
a commercial standard sample of HMF (Sigma, St. Louis,
MO, USA).
Determination of browning degree (BD)
The method was proposed by Liu et al. [13]. The BD of the
samples was measured by their absorbance at 420 nm with
spectrophotometer (UV-762, Lingguang, Shanghai, China)
using a 10 mm path length cell.
Determination of sucrose, glucose, and fructose
A Knauer HPLC system (Knauer Co. Ltd., German)
was used, equipped with a pump (K-1001) connected a
refractive index detector (RI-2301) and a 20 µl loop. The
column was a Waters Sugar pak-1 (6.5×300 mm i.d.)
(Waters Co., USA). The mobile phase was de-ionized
water; the flow rate was 0.5 ml min−1 at 90 ◦ C. The samples
prepared from the determination of BD. Quantification
were carried out by the external standard method, using
284

a commercial standard of sucrose, glucose, and fructose tionmeter (Shanghai Precision & Scientific Instrument Co.,
(Sigma, St. Louis, MO, USA). Shanghai, China) at 20 ◦ C.

Determination of total amino acid (TAA) Determination of L-ascorbic acid

Chromatographic analysis was proposed by Waters [29].
HPLC system (Waters, USA) composed of 626 LC system
with heater, 717 plus auto-sampler, 474 scanning fluores-
cence detector (at 250 nm excitation, 395 nm emission),
and AccQ.Tag column (4.6×150 mm i.d.) equipped with a
Nova-pak C18 (Waters) guard column.
Hundred microliter of sample was filled into a 16×25 mm
test tube and 5 ml, 6 mol l−1 HCl. Thoroughly flushed, the
tube with nitrogen or argon was capped and placed in an
oven at 112 ◦ C for 22 h. The internal standard (10 ml of
2.5 mM alpha-aminobutyric acid) was added to the cooled
hydrolysate mixture, transferred to a 250 ml volumetric
flask, and filled to the meniscius with de-ionized water. A
0.5 ml of the solution was filtered with a 0.45 µm acid-
compatible filter.
Ten microliter of filtrate was mixed with 70 µl of
AccQ.Fluor derivatization buffer, and 20 µl of AccQ.Fluor
reagent was added to derivatize. The sample was trans-
ferred to a limited volume insert in an autosampler vial and
heated at 55 ◦ C for 10 min in a heating block.
Injection volume was 5 µl. The column temperature was
37 ◦ C, the flow rate was 1.0 ml min−1 , and the composi-
tion of the mobile is shown in Table 1. Quantification was
carried out by the internal standard method using commer-
cial standard L-amino acid and alpha-aminobutyric acid
(Sigma, St. Louis, MO, USA).
L-Ascorbic acid concentration was determined using Food
and Agriculture Organization (FAO) method employing
2,6-dichloindophenol potentiometricatitration procedure
[30].
Data analysis
The data were subjected to regression analysis using the
first-order reaction (Eq. 1). The kinetic rate constants were
calculated from the regression of the different index values
vs. storage time.
A = A0 exp(kt) (1)
where A and A0 are the values of the index at time t and 0,
respectively; k is the kinetic rate constant, and t the time.
Each experiment was performed in triplicates. The data
were the mean values expressed as milligram per liter at 8.0
◦
Brix. Results are expressed or plotted as the mean value
± standard error. One-way analysis of variance (ANOVA)
was performed to evaluate the influence of the storage con-
ditions on HMF accumulation and the degree of browning
with the software SAS 9.0 professional (SAS Institute Inc.,
Cary, NC, USA). The Student–Newman–Keuls test was
used to estimate the statistically significant differences.

Determination of pH Results and discussion

pH was measured at 20 ◦ C with a pH meter (Thermo Orion Change of TS, sucrose, glucose, fructose, and TAA
868, USA), which was calibrated with pH 4.0 and 7.0 in stored CJC
buffers.
Table 2 gives the change of the major constituents includ-
Determination of total soluble solids (TSS) ing sucrose (which could invert into glucose and fructose),
glucose, fructose, and TAA in CJC correlated with Mail-
The soluble solids content of juice concentrates was deter- lard reaction. After 150-day storage at 25 and 37 ◦ C, TS,
mined as degree Brix using WAY-2S digital Abbe Refrac- sucrose, and TAA were significantly decreased, while the
glucose and the fructose were significantly increased. Dur-
Table 1 Flow rate and mobile phase constituent in TAA determi- ing storage, two chemical reactions occurred, including the
nation inversion reaction of sucrose and Maillard reaction. Gen-
Time (min) Aa (%) Bb (%) Cc (%) erally, the glucose and fructose should be reduced during
0 100 0 0 Maillard reaction, but they increased in our observation.
0.5 99 1 0 The unconformity of glucose and fructose was caused by
18 95 5 0
sucrose inversion and Maillard reaction during storage, in-
19 91 9 0
dicating that reaction rate of sucrose inversion was greater
than that of Maillard reaction. The decrease in sucrose was
28 83 17 0
attributed to the sucrose inversion into glucose and fructose
35 0 60 40
at higher temperature and at lower pH, while the decrease in
38 100 0 0
TS resulted from Maillard reaction. Similarly, the decrease
a
A, AccQ.Tag eluent A of the TAA was caused by Maillard reaction.
b
B, acetonitrile After 150-day storage at 0 and −18 ◦ C, TS, sucrose,
c
C, water glucose, fructose, and TAA had less significant change.
Table 2 Major constituents in CJC stored at 0 day and after 150 days with Maillard reaction (expressed as gram per liter at 8.0 ◦ Brix)
◦
Brix Ta (◦ C) Sb Fc Gd TSe TAAf
0 day 150 days 0 day 150 days 0 day 150 days 0 day 150 days 0 day 150 days
20 −18 23.41±0.41 21.81±0.07 12.15±0.11 12.18±0.22 12.20±0.21 12.36±0.34 47.77±0.13 46.35±1.16 1.45±0.05 1.15±0.02
0 23.41±0.41 20.10±0.20 12.15±0.11 12.16±0.33 12.20±0.21 13.35±0.22 47.77±0.13 45.61±0.67 1.45±0.05 1.06±0.01
25 23.41±0.41 11.81±0.28 12.15±0.11 17.42±0.09 12.20±0.21 15.43±0.12 47.77±0.13 44.66±0.45 1.45±0.05 0.68±0.01
37 23.41±0.41 2.06±0.15 12.15±0.11 22.14±0.03 12.20±0.21 20.08±0.09 47.77±0.13 44.28±0.59 1.45±0.05 0.55±0.01
40 −18 23.08±0.61 21.07±0.30 12.17±0.13 12.48±0.27 12.03±0.55 12.37±0.16 47.27±0.52 45.92±0.30 1.35±0.02 1.23±0.01
0 23.08±0.61 19.55±0.52 12.17±0.13 12.81±0.23 12.03±0.55 13.05±0.16 47.27±0.52 45.41±0.67 1.35±0.02 1.14±0.02
25 23.08±0.61 10.28±0.42 12.17±0.13 18.60±0.63 12.03±0.55 15.52±0.57 47.27±0.52 44.40±1.06 1.35±0.02 0.61±0.02
37 23.08±0.61 0.92±0.09 12.17±0.13 22.06±0.14 12.03±0.55 20.61±0.39 47.27±0.52 43.59±0.71 1.35±0.02 0.41±0.02
60 −18 22.61±0.63 20.76±0.54 12.22±0.22 12.08±0.14 12.11±0.48 13.07±0.47 46.94±0.24 45.19±0.63 1.26±0.02 1.13±0.01
0 22.61±0.63 19.18±0.42 12.22±0.22 12.98±0.61 12.11±0.48 12.36±0.51 46.94±0.24 45.25±1.16 1.26±0.02 1.12±0.01
25 22.61±0.63 10.28±0.76 12.22±0.22 17.18±0.17 12.11±0.48 16.74±0.23 46.94±0.24 44.20±1.17 1.26±0.02 0.60±0.02
37 22.61±0.63 0.89±0.05 12.22±0.22 21.16±0.22 12.11±0.48 18.74±0.31 46.94±0.24 40.79±0.65 1.26±0.02 0.39±0.01
a
T, temperature
b
S, sucrose
c
F, fructose
d
G, glucose
e
TS=S+F+G
f
TAA, total amino acid
285
286

3.8 a 30
37 ˚C 0.8
-18 ˚C 0 ˚C 25 ˚C 20 ˚ Brix
25
-18 ˚C
3.7 0 ˚C
20 25 ˚C 0.6
37 ˚C

[HMF] (mg L )
-1
3.6 15 -18 ˚C
pH value

[BD]
0 ˚C 0.4
25 ˚C
10
37 ˚C
3.5
5 0.2

3.4 0
0.0
0 30 60 90 120 150
storage time (d)
3.3
20 40 60
b 80 1.5
TSS (˚Brix)
40 ˚Brix
Fig. 2 Change of pH value after 150-day storage of CJC 70
-18 ˚C
0 ˚C 1.2
60
25 ˚C

[HMF] (mg L )
Change of pH in stored CJC

-1
50 37 ˚C
-18 ˚C 0.9

[BD]
40 0 ˚C
The pH change after 150-day storage at different temper-
25 ˚C
ature is shown in Fig. 2. After 150-day storage at 25 and 30
37 ˚C 0.6
at 37 ◦ C, the pH decrease was observed; the higher the 20
storage temperature and the soluble solid content were, the 0.3
10
greater the decrease of pH. This observation was similar
to previous studies. Beck et al. reported that the decrease 0
0.0
in pH observed during the Maillard reaction could be at- 0 30 60 90 120 150
tributed to the reaction of amines to form compounds of storage time (d)
lower basicity and to the degradation of sugars into acids
[31]. Trifiro et al. and Carabasa-Giribet et al. found that c 240 3.6
60 ˚Brix
the decrease in pH was due to the condensation between 210
-18 ˚C 3.0
free amino of amino acids and carbonyl groups of glucose, 180 0 ˚C
respectively [32, 33]. However, after 150-day storage at 0 25 ˚C
2.4
and −18 ◦ C, the pH did not change, indicating that lower 150 37 ˚C
[HMF] (mg L-1)

-18 ˚C
temperature of storage has less effect on pH of CJC. 120 0 ˚C 1.8

[BD]
The decrease of pH at higher temperature storage would 90
25 ˚C
push the formation of HMF. The HMF formation pathway 37 ˚C 1.2

predominates at a lower pH [34]. Shallenberger and Mattick 60

0.6
found the HMF formation doubled at pH 3 as compared to 30
pH 4.6, Apriyantono and Ames showed that a higher pH 0.0
0
did lower HMF concentrations [35, 36].
0 30 60 90 120 150
storage time (d)

Kinetic analysis of HMF formation and BD
Studies showed that the formation of HMF resulted from
Maillard reaction, L-ascorbic acid decomposition, and
caramilization [37–41]. In this study, L-ascorbic acid was
not detected in CJC, and the storage temperature was less
than 40 ◦ C. The decomposition of L-ascorbic acid and the
caramilization reaction could not occur. Thus, the forma-
tion of HMF in CJC was caused by the Maillard reaction
between amino acids and reducing sugar.
The first-order reaction of Maillard reaction in a glucose–
lysine model system was reported by Cerrutti et al. [42].
Also, Toribio et al. have revealed that this reaction follows
a first-order reaction in apple juice concentrates during
storage [43]. In this study, non-enzymatic browning was
Fig. 3 (a) HMF formation and BD of 20 ◦ Brix CJC stored for 150
days. Solid line represented HMF, dash line represented BD. [HMF],
HMF content; [BD], browning degree value (absorbance at 420 nm).
(b) HMF formation and BD of 40 ◦ Brix CJC stored for 150 days.
Solid line represented HMF, dash line represented BD. [HMF], HMF
content; [BD], browning degree value. (c) HMF formation and BD of
60 ◦ Brix CJC stored for 150 days. Solid line represented HMF, dash
line represented BD. [HMF], HMF content; [BD], browning degree
value
considered as the Maillard reaction, the first-order reaction
was used in fitting to the experimental data.
Fig. 3a–c shows the effect of storage time, storage tem-
perature, and soluble solids on HMF formation and BD in
CJC expressed at 8 ◦ Brix. One-way ANOVA was applied
to see the net effect of temperature on HMF formation and
BD for 150-day storage at −18, 0, 25, and 37. It showed
 287

that lower temperatures such as −18 and 0 ◦ C had no sig- a

nificant effect on HMF formation and BD, while higher 25 20 ˚ Brix
temperatures such as 25 and 37 ◦ C significantly increased
the HMF and BD at the same storage time and the same 20
concentration of soluble solid (p<0.01). The storage time
and the soluble solid content also had a significant effect

[HMF] (mg L )
15
on HMF and BD at 25 ◦ C or 37 ◦ C (p<0.01). The change

-1
of BD was similar to the change of HMF formation, which
was illustrated in Fig. 3a–c. 10 37 ˚ C
The experimental data in Fig. 3a–c were fitted to Eq. [1]. 25 ˚ C
The kinetics parameters such as the rate constant k1 of HMF 5
formation and k2 of BD at 25 ◦ C or 37 ◦ C and their regres-
sion coefficients R2 were calculated and are listed in Table
0
3. As the storage temperature at the same concentration of
soluble solid or the concentration of soluble solid at the 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8
same storage temperature increased, the rate constant k1 of
[BD]
HMF formation and k2 of BD also increased, implying that
the increment of the storage temperature or the concentra- b 80
tion of soluble solid resulted in the increase of HMF content 40 °Brix
and BD. Moreover, all the regression coefficients R2a and
R2b were greater than 0.944 and 0.908, respectively, indicat- 60
ing that the HMF formation and BD followed a first-order
reaction well at higher temperature such as 25 ◦ C or 37 ◦ C. -1
mg L
These results were consistent with Toribio et al. [43] and 40
Johnson et al. [44]. However, the regression coefficients at
lower temperature such as −18 and 0 ◦ C was smaller (not 25 °C
[HMF

shown), indicating that the HMF formation and BD did not 20 37 °C

conform to the first-order reaction at −18 and 0 ◦ C.

0
The relationship between HMF and BD (A420)
0.0 0.3 0.6 0.9 1.2 1.5 1.8
HMF and BD were two quality indicators in fruit and veg- [BD]
etable juice. As shown in Fig. 4a–c and Table 4, there is
a linear regression of HMF content and BD in CJC sam- c 250
ples (n=126), suggesting a significant correlation between 60 ˚ Brix
HMF and BD. The equation in Table 4 could generally be
expressed as Eq. [2]. It was found that the parameters such 200
c and d in Eq. 2 had a close correlation with the total soluble
solid [TSS] depending on the storage temperature.
150
[HMF]( mg L )
-1

[HMF] = c + d × [BD] (2)

100
25 ˚C
Table 3 Rate of HMF formation and BD in CJC stored for 150 37 ˚C
days
◦ 50
Brix Ta (◦ C) HMFb BDc
k1 (mg L−1 d−1 ) R2a k2 (mg L−1 d−1 ) R2b
20 25 0.0119 0.990 0.0058 0.908 0
37 0.0163 0.964 0.0096 0.960 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0
40 25 0.0138 0.964 0.0067 0.927 [BD]
37 0.0178 0.975 0.0111 0.973
Fig. 4 (a) Correlation between HMF formation and BD in 20 ◦ Brix
60 25 0.0150 0.952 0.0088 0.937 CJC. [HMF], HMF content; [BD], browning degree value. (b) Cor-
37 0.0173 0.944 0.0136 0.962 relation between HMF formation and BD in 40 ◦ Brix CJC. [HMF],
a HMF content; [BD], browning degree value. (c) Correlation between
T, temperature HMF formation and BD in 60 ◦ Brix CJC. [HMF], HMF content;
b
HMF, 5-hydroxymethylfurfural [BD], browning degree value
c
[BD], browning degree
288

Table 4 The relationship between HMF and BD

Conclusion
T (◦ C) ◦
Brix Equation R2
25 20 [HMF[=−1.0291+9.3384×[BD] 0.908 Higher temperature at 25 and 37 ◦ C had a significant effect
40 [HMF]=−3.2835+17.6422×[BD] 0.958 on TS, sucrose, glucose, fructose, TAA, pH, HMF, and BD,
60 [HMF]=−5.5265+23.2451×[BD] 0.976 while lower temperature at 0 and −18 ◦ C had less effect.
37 20 [HMF]=−6.4822+36.6814×[BD] 0.968 The storage time and the soluble solid of CJC stored at 25
40 [HMF]=−15.8461+56.2109×[BD] 0.976 and 37 ◦ C also had a significant effect on HMF formation
60 [HMF]=−23.2138+65.6758×[BD] 0.987 and BD. At 25 and 37 ◦ C, the formation of HMF and BD
followed a first-order reaction well as a function of storage
[HMF], HMF content; [BD], browning degree value time; a good correlation between HMF formation and BD
occurred. The formation of HMF could described by the
total soluble solid value ([TSS]) and the BD value ([BD],
absorbance at 420 nm) of CJC, the equation was [HMF]
When the storage temperature was 25 ◦ C, c and d could be (content of HMF) = 1.2177 − 0.1124 × [TSS] + (2.8350
described by the total soluble solid [TSS] such as Eqs. (3) + 0.3477 × [TSS]) × [BD] and [HMF] = 1.5510 − 0.4813
and (4). ×[TSS] + (23.8847 + 0.7249 ×[TSS]) × [BD] at 25 and
37 ◦ C, respectively.
c = 1.2177 − 0.1124 × [TSS], R2
Acknowledgement This research work was funded by the National
= 0.999, p < 0.0001, (3) Agro-processing Project in the Tenth Five Year Plan of P.R. China
(No. 2001BA501A23).

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