Food Research International 49 (2012) 439–445

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Food Research International

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Effects of sugars in batter formula and baking conditions on 5-hydroxymethylfurfural

and furfural formation in sponge cake models
Yu-Yu Zhang, Yi Song, Xiao-Song Hu, Xiao-Jun Liao, Yuan-Ying Ni, Quan-Hong Li ⁎
College of Food Science and Nutritional Engineering, China Agricultural University National Engineering Research Center for Fruits and Vegetables Processing, China
Key Laboratory of Fruit and Vegetable Processing, Ministry of Agriculture, Beijing 100083, China

a r t i c l e i n f o a b s t r a c t

Article history: 5-Hydroxymethylfurfural (HMF) and furfural (F) could be formed in sugar-rich foods during baking. The
Received 20 March 2012 effects of batter formula and baking conditions on the formation of HMF and F and the kinetics of HMF for-
Accepted 10 July 2012 mation during baking were studied in 15 sponge cake models. A fixed amount of sucrose (40%) was included
as minimum in sponge cake models to improve the batter consistency and texture of sponge cakes. The
Keywords:
results showed that increase of the sugar and citric acid amount in the batter formula increased HMF and F
5-Hydroxymethylfurfural
Furfural
formation during baking at 205 °C for 11 min. With increasing of the baking temperature and time, the con-
Cake centrations of HMF and F were also increased in the sponge cake models, and the maximum concentrations of
Baking HMF and F attained were 4100 ± 11 mg kg−1 and 59 ± 1 mg kg−1, respectively. Formation of HMF in the
Formation sponge cake models (Models 11–15) followed a first-order kinetics. By applying the Arrhenius equation,
Kinetics the activation energy (Ea) of sponge cake models was found to a span range between 15.4 kJ mol −1 and
25.8 kJ mol −1. The results showed that sugar type, pH and baking temperature and time strongly affected
HMF and F formation in sponge cake models.
© 2012 Elsevier Ltd. All rights reserved.

1. Introduction reducing carbohydrates directly suffer 1,2-enolisation, dehydration

Bakery products, such as biscuit, cookie and cakes, constitute one
of the most consumed foods in the world. Among them, cakes are
particularly popular and associated in the consumer's mind with a
delicious product. The major ingredients used in the preparation of
sponge cakes are wheat flour, egg and sugar. Sugar not only imparts
sweetness, but contributes to the fresh flavor quality of cakes during
thermal processing and acts as a tenderizer by retarding and
restricting gluten formation (Heenan, Dufour, Hamid, Harvey, &
Delahunty, 2010). The cake batters in which the sucrose is replaced
with different sugars result in cakes with different cake structure
and cake volume (Ikawa, 1998).
Sponge cake batter is conventionally baked at high temperature
(about 200 °C). Baking is a complex process in which chemical and
physical changes take place simultaneously. During baking, the
Maillard reaction and caramelization are responsible for the forma-
tion of desirable taste and color of baked products and furans, such as
5-hydroxymethylfurfural (HMF) and furfural (F) (Kroh, 1994). After
hydrolysis of sucrose at high temperature, the sugar decomposes into
furans by two possible pathways: caramelization and Maillard reaction
(Ameur, Trystram, & Birlouez-Aragon, 2006). In caramelization, the
⁎ Corresponding author at: College of Food Science and Nutritional Engineering, China
Agricultural University, Beijing 100083, China. Tel.: +86 10 62738831; fax: +86 10
62737761.
E-mail address: quanhong_li@hotmail.com (Q.-H. Li).
and cyclization reactions (Kroh, 1994). In the Maillard reaction, the
Amadori product is subjected to enolization and subsequent dehydra-
tion of the sugar moiety and the release of an intact amino acid
(Hodge, 1953). The Maillard reaction is favored in foods with a high
protein and carbohydrate content, intermediate moisture content, tem-
peratures above 50 °C and a pH of 4–7, but the caramelization reaction
needs more drastic conditions namely higher temperatures (>120 °C),
pH and low water activity (Gökmen, Açar, Serpen, & Morales, 2008;
Kroh, 1994). The formation of HMF and F is influenced by the concentra-
tions and type of sugar and is favored by lower pH and low water activ-
ity (Ameur et al., 2006; Gökmen, Açar, Köksel, & Acar, 2007). In acidic
medium, HMF and F are formed respectively by the decomposition
of hexoses and pentoses during heating, after a slow enolization and
a fast β-elimination of three water molecules (Belitz, Grosch, &
Schieberle, 2009; Kroh, 1994; Pereira, Albuquerque, Ferreira, Cacho, &
Marques, 2011). HMF and F have been evaluated as indicators of the
quality of thermal processing in foods including fruit juices, biscuits,
breakfast cereals and baby foods (Delgado-Andrade, Rufián-Henares,
& Morales, 2009; Gökmen & Acar, 1999; Gökmen & Şenyuva, 2006;
Rufián-Henares, Delgado-Andrade, & Morales, 2006). Along with the
Maillard reaction and caramelization, the formation of HMF and F are
particularly interesting because of their accumulation during the baking
process.
HMF is not present in fresh, untreated foods, but it rapidly accumu-
lates in sugar-rich foods during heating at high temperature (Ameur
et al., 2006). The accumulation of HMF is considered undesirable in

0963-9969/$ – see front matter © 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodres.2012.07.012
440 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445

thermally processed foods, and its presence in food is focused on some Table 1
potential toxicological concern (Abraham et al., 2011). The toxicolog- The composition of models used to prepare sponge cakes.

ical relevance of HMF still remains unclear, and the subject of in vitro Model Ingredients (%)a
studies on genotoxicity and mutagenicity is still a matter of debate
Wheat Egg Sucrose Glucose Fructose Lactose Maltose Citric
(Cuzzoni, Stoppini, Gazzani, & Mazza, 1988; Janzowski, Glaab, flour acid
Samimi, Schlatter, & Eisenbrand, 2000; Surh & Tannenbaum, 1994).
1 100 100 0–120b 0 0 0 0 0
From a safety perspective and for food quality assurance, HMF legal 2 100 100 40 0–80c 0 0 0 0
limits were already issued for some foodstuffs, namely for concentrat- 3 100 100 40 0 0–80c 0 0 0
ed rectified grape must: EC Regulation No. 1493/99 sets up a limit of 4 100 100 40 0 0 0–80c 0 0
25 ppm (Abraham et al., 2011; Falcone, Tagliazucchi, Verzelloni, & 5 100 100 40 0 0 0 0–80c 0
6 100 100 120 0 0 0 0 0–6.0d
Giudici, 2010). 7 100 100 40 80 0 0 0 0–6.0d
The kinetic formation of HMF during heating has been studied in 8 100 100 40 0 80 0 0 0–6.0d
rice wine (Chen, Huang, Liu, Peng, & Huang, 2010), unifloral honey 9 100 100 40 0 0 80 0 0–6.0d
(Fallico, Zappalà, Arena, & Verzera, 2004) and cookie model system 10 100 100 40 0 0 0 8.0 0–6.0d
11 100 100 120 0 0 0 0 0
(Ameur, Mathieu, Lalanne, Trystram, & Birlouez-Aragon, 2007; Ameur
12 100 100 40 80 0 0 0 0
et al., 2006), showing the influences of baking temperature, time and 13 100 100 40 0 80 0 0 0
type of sugar. Ameur et al. (2007) reported that the accumulation of 14 100 100 40 0 0 80 0 0
HMF followed a first-order kinetics, highly depending on the baking 15 100 100 40 0 0 0 80 0
temperature and type of sugar, and sucrose-cookie produced less HMF The initial moisture content of cake batter is 22.5 ± 0.4%.
than glucose-cookie or fructose-cookie at below 250 °C. However, less With adding citric acid 0, 2.0, 4.0, or 6.0%, the pH of cake batter is 7.5 ± 0.3, 4.3 ± 0.5,
information is available for the effects of the type and concentration of 3.7 ± 0.5 and 3.4 ± 0.2, respectively.
a
Baker's percentages.
sugars, pH and moisture content on the formation of HMF and F in b
0, 40, 60, 80, 100, or 120%.
sponge cakes. c
0, 20, 40, 60, or 80%.
The aim of this work was to investigate the effects of batter formula d
0, 2.0, 4.0, or 6.0%.
with different pH and concentrations of sugars (sucrose, glucose, fruc-
tose, lactose and maltose) and the baking process (temperature and
time) on HMF and F formations in sponge cake model systems. Kinetic it became foamy and formed stiff peaks. Then flour was added and gently
models for HMF formation in sponge cake models under the heating folded into the whipped batter in small increments using a plastic kitchen
treatment were also determined. The changes of moisture content spatula. A portion of the batter (10.0±1.0 g) was placed into an alumi-
and pH of sponge cake models were studied to discuss their effect on num foil baking cup with a diameter of 3 cm and a height of 1 cm and
HMF and F formation during baking. then baked. Two cakes were produced from each cake model system.
Batters of Models 1–10 were baked at 205 °C for 11 min (Gökmen
2. Materials and methods et al., 2007). Batters from Models 11–15 were baked at different tem-
peratures and times (8, 10, 12, 15 and 20 min at 220 °C and 210 °C;
2.1. Materials 10, 12, 15, 20 and 25 min at 200 °C; 10, 12, 15, 20, 25 and 30 min at
190 °C; 10, 12, 15, 20, 25, 30 and 35 min at 180 °C).
High purity (>99%) HMF (5-hydroxymethylfurfural) standard was
purchased from Sigma-Aldrich (St. Louis, MO, USA). Wheat flour (8.5% 2.3. Measurement of pH
protein; 12.7% moisture content; 0.5% ash; 0.2% soluble reducing sugar)
was supplied by Damofang (Beijing Damofang Flour Co., Ltd, China). A ground sample (0.4 g) of cake was mixed with 20 mL of water
Fresh egg was purchased from local supermarkets. The sucrose, glucose, and vortexed for 3 min. The mixture was held at ambient tempera-
fructose, lactose, maltose, zinc sulfate and K4Fe(CN)6 (all AnalaR grade) ture for 1 h to separate solid and liquid phases. The pH of the super-
were purchased from Beijing Chemical Works (China). Methanol (HPLC natant was measured using a Thermo Orion 868 pH meter (Thermo
grade) was purchased from Fisher Scientific (Fair Lawn, New Jersey, Fisher Scientific, Inc., Pittsburgh, PA).
USA). The ultra-pure water (18.25 mΩ) was prepared in a Millipore
apparatus (Simplicity, Canada). 2.4. Measurement of moisture
Carrez Ι solution was prepared by dissolving 15 g of potassium
hexacyanoferrate in 100 mL of water, and Carrez ΙΙ solution by dis- The moisture content of the cakes was measured using an electronic
solving 30 g of zinc sulfate in 100 mL of water (García-Villanova, moisture analyzer (Sartorius Moisture Analyzer 150, Germany). The
Guerra-Hernández, Martínez-Gómez, & Montilla, 1993). mean of two measurements was reported.

2.2. Preparation of sponge cakes 2.5. HMF and F measurements

The sponge cake models were prepared according to a recipe de-
scribed by Gómez, Ruiz, and Oliete (2011), and Matsakidou, Blekas,
and Paraskevopoulou (2010), with some modifications, in order to
study the effects of type and amount of sugar (Models 1–5), the effect
of amount of citric acid (Models 6–10) and baking conditions (Models
11–15) on HMF and F formation. Sponge cakes were prepared by
wheat flour, whole egg, sugar and citric acid, and the composition of
models is represented in Table 1.
The ingredients were thoroughly mixed into a cake batter according
to these models, and then baked in an oven (BEC-OVST 03, Baker's
Kingdom Shanghai Co., Ltd., China). Whole egg and sugar were mixed
with a whip at 90 rpm for 5 min with a kitchen mixer (HK-TU, Baker's
Kingdom Shanghai Co., Ltd., China), and then at 150 rpm for 7 min until
HMF and F were measured using the method of Gökmen and
Şenyuva (2006) and Gökmen et al. (2008) with some modifications.
Sponge cake samples were ground and stored at − 18 °C prior to
analysis. A fraction sponge cake sample (1 g) was weighed into a
20 mL centrifuge tube with a cap. A total of 250 μL of Carrez Ι and
250 μL of Carrez ΙΙ solutions were added, and the volume was com-
pleted to 10 mL with deionized water. The centrifuge tube was
then shaken vigorously for 3 min, and centrifuged for 10 min at
10,000 × g, at 4 °C. The solution was filtered through a 0.45 μm
disk filter, and stored at − 18 °C until analysis. All the experiments
were carried out in duplicate.
A Shimadzu model HPLC system (Shimadzu Corp., Kyoto, Japan)
was employed for the HMF and F analysis. The chromatography
 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445 441

a
Detector A: 13.840 min 15.132 min
Standard HMF
284nm F
7.5

mV
5.0

2.5

0.0
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0
Retention time (min)
b
Detector A:
10.0 Sample
284nm
7.5
mV

5.0 13.920 min 15.101 min

HMF F
2.5

0.0
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0
Retention time (min)

Fig. 1. HPLC spectrogram (284 nm) of 5-hydroxymethylfurfural (HMF) and furfural (F) for the standard solution (a) and the sponge cake model sample (b).

column (Venusil XBP-C18 (250 mm× 4.6 mm i.d., 5 μm particle size)) 2.6. Kinetic and statistical analysis
was obtained from Bonna-Agela Technologies Inc. (Venusil, Agela,
USA). Data were collected, processed, and analyzed using Class-VP Chro- Kinetic analysis was according to the method described by Chen et
matography Laboratory Automated software (Shimadzu Corp.). The al. (2010). In the HMF formation reaction, the product may increase
chromatographic separations were carried out at 284 nm and 30 °C. linearly with time (zero-order kinetics) or exponentially with time
The mobile phase was methanol–water (5:95, v:v) with a flow of (first-order kinetics) (Garza, Ibarz, Pagán, & Giner, 1999).
1.0 mL min−1, and the injection volume was 20 μL. The concentration
of HMF was determined using an external calibration curve. Calibration C ðt Þ ¼ C 0 þ k⋅t ðzero  order kineticsÞ ð1Þ
standard solutions ranging from 0.0143 to 11.44 μg mL −1 (HMF) and
0.0131 to 10.48 μg mL−1 (F) were prepared. C ðt Þ ¼ C 0 expðk⋅t Þ ðfirst  order kineticsÞ ð2Þ

a b
70
2.4
60 Sucrose (Model 1) Sucrose (Model 1)
HMF content (mg kg-1)

2.0
Glucose (Model 2) Glucose (Model 2)
F content (mg kg-1)

50
Fructose (Model 3) 1.6 Fructose (Model 3)
40 Lactose (Model 4) Lactose (Model 4)
Maltose (Model 5) 1.2 Maltose (Model 5)
30

20 0.8

10 0.4

0 0.0

0 20 40 60 80 100 120 0 20 40 60 80 100 120

Sugar amount in the model (%) Sugar amount in the model (%)

c d
1800 Sucrose (Model 6) 40 Sucrose (Model 6)
Glucose (Model 7) Glucose (Model 7)
HMF content (mg kg-1)

1500 Fructose (Model 8) 32 Fructose (Model 8)

F content (mg kg-1)

Lactose (Model 9) Lactose (Model 9)

1200
Maltose (Model 10) 24 Maltose (Model 10)
900
16
600
8
300

0 0

0 1 2 3 4 5 6 0 1 2 3 4 5 6
Citric acid amount in the model (%) Citric acid amount in the model (%)

Fig. 2. Effects of the amount of sugars and citric acid on the formation of 5-hydroxymethylfurfural (HMF, a and c) and furfural (F, b and d) in sponge cakes upon baking at 205 °C for
11 min.
442 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445

8.0 220 oC 3.1. Effects of batter formula

Model 11
210 oC
Replacing all of the sucrose with other sugars in the sponge cake
7.5 200 oC model systems adversely affected the sponge cake structure during
190 oC the course of preliminary baking trials. Therefore, a fixed amount
180 oC of sucrose (40%) was necessarily included as minimum in all cake
7.0 models to improve the batter consistency and texture of cakes.
Sponge cakes were baked at 205 °C for 11 min, as in the method
pH

described by Gökmen et al. (2007).

6.5
The type of sugar added in addition to the sucrose base showed a
strong influence on HMF and F formation in sponge cakes (Fig. 2).
Sucrose, lactose and maltose yielded less HMF than did glucose and
6.0
fructose (Fig. 2a). However, maltose yielded more F than other sugars
(Fig. 2b). Upon baking at 205 °C for 11 min, 0.4 ± 0.01 mg kg −1 of
5.5 HMF and 0.1 ± 0.01 mg kg −1 of F formed in sponge cakes with 40%
0 5 10 15 20 25 30 35 40 sucrose (Fig. 2a). However, replacing a portion of the sucrose with
Baking time (min) glucose, fructose, lactose and maltose in the cake models resulted in

Fig. 3. Changes of pH in the sponge cake Model 11 during baking.

Table 2
Furfural concentration in sponge cake models at different baking temperature and time.

Time Furfural (mg kg−1)

where, C0 is the initial concentration of HMF (mg HMF per kg cake), (min)
180 °C 190 °C 200 °C 210 °C 220 °C
C(t) the HMF concentration (mg HMF per kg cake); t the baking
time (min); and k the rate constant (min −1). Model 11
8 – – – 0.2 ± 0.0 a 0.6 ± 0.0 bc
Regression analysis (curve fitting) and the calculation of kinetic 10 0.2 ± 0.0 a 0.2 ± 0.0 a 0.2 ± 0.0 a 0.5 ± 0.1 abc 1.3 ± 0.1 d
rate constant were performed using the Microcal Origin 8.0 software. 12 0.2 ± 0.0 a 0.2 ± 0.0 a 0.4 ± 0.0 ab 0.8 ± 0.1 c 1.5 ± 0.2 d
Analysis of variance (ANOVA), as confirmatory analysis, was per- 15 0.3 ± 0.0 ab 0.4 ± 0.0 ab 0.8 ± 0.1 c 2.3 ± 0.1 e 16.3 ± 0.3 j
formed and then the means of results were compared by Duncan's 20 0.3 ± 0.1 ab 0.8 ± 0.1 c 2.7 ± 0.3 f 24.1 ± 0.2 k 36.4 ± 0.3 m
25 1.6 ± 0.2 d 4.8 ± 0.1 h 12.3 ± 0.1 i – –
multiple range tests in the SPSS software. All determinations were
30 4.0 ± 0.6 g – – – –
carried out in triplicate and results were expressed as the mean 35 25.4 ± 0.3 l – – – –
value ± standard deviation.
Model 12
8 – – – 1.0 ± 0.1 bc 2.3 ± 0.1 f
10 0.4 ± 0.0 a 0.6 ± 0.1 ab 0.7 ± 0.1 ab 1.7 ± 0.2 e 3.0 ± 0.4 gh
3. Results and discussion 12 0.5 ± 0.0 a 1.2 ± 0.1 cd 2.7 ± 0.0 fg 3.2 ± 0.1 hi 3.6 ± 0.3 i
15 0.7 ± 0.1 ab 1.3 ± 0.2 cde 3.0 ± 0.1 gh 4.1 ± 0.1 j 5.5 ± 0.1 lm
Fig. 1 depicts the chromatographic separation of HMF and F in 20 1.6 ± 0.2 e 4.1 ± 0.1 j 6.9 ± 0.1 n 18.6 ± 0.1 o 59.0 ± 1.0 q
standard solution and the sponge cake model sample. It could be seen 25 3.0 ± 0.1 gh 5.1 ± 0.2 l 25.9 ± 0.5 p – –
30 4.6 ± 0.3 k – – – –
from the chromatogram that both HMF and F were completely resolved
35 5.6 ± 0.1 m – – – –
from sponge cake model samples. The limit of detection (LOD) and the
limit of quantification (LOQ) were calculated as follows: 3.3 σ/b and Model 13
10 σ/b, respectively, where σ is the y-intercept standard deviation and 8 – – – 0.1 ± 0.0 a 1.2 ± 0.0 c
10 0.2 ± 0.0 ab 0.3 ± 0.0 ab 0.3 ± 0.0 ab 0.3 ± 0.0 ab 10.3 ± 0.1 i
b is the slope of the linear regression (Pereira et al., 2011). The obtained
12 0.3 ± 0.0 ab 0.4 ± 0.0 ab 0.5 ± 0.0 b 1.5 ± 0.1 d 12.2 ± 0.1 j
LOD value and LOQ value were 0.1 mg kg−1 and 0.3 mg kg−1. 15 0.3 ± 0.0 ab 0.5 ± 0.0 b 1.5 ± 0.1 d 4.4 ± 0.1 f 29.4 ± 0.1 m
20 0.5 ± 0.0 b 1.7 ± 0.1 d 3.4 ± 0.1 e 13.4 ± 0.1 k 40.6 ± 0.2 n
25 4.8 ± 0.1 g 5.8 ± 0.4 h 12.0 ± 0.1 j – –
30 13.3 ± 0.6 k – – – –
– – – –
25 Model 11 220 oC 35 27.5 ± 0.3 l

210 oC Model 14
8 – – – 0.4 ± 0.0 a 2.7 ± 0.0 cd
20 200 oC 10 0.3 ± 0.0 a 1.4 ± 0.1 b 2.1 ± 0.1 bc 2.1 ± 0.2 bc 3.5 ± 0.1 e
190 oC 12 0.4 ± 0.0 a 2.7 ± 0.3 cd 3.2 ± 0.2 de 3.4 ± 0.1 de 4.4 ± 0.2 f
Moisture content (%)

15 0.4 ± 0.0 a 3.1 ± 0.4 de 8.6 ± 0.1 i 9.2 ± 0.3 i 13.5 ± 0.6 k
15 180 oC 20 1.9 ± 0.1 b 5.1 ± 0.1 g 19.3 ± 0.1 m 28.7 ± 1.5 n 37.3 ± 0.1 o
25 5.1 ± 0.3 g 12.5 ± 0.2 j 28.1 ± 1.0 n – –
30 6.0 ± 0.1 h – – – –
10 35 16.8 ± 0.1 l – – – –

Model 15
8 – – – 0.7 ± 0.1 ab 2.1 ± 0.1 c
5
10 0.1 ± 0.0 a 0.1 ± 0.0 a 2.1 ± 0.3 c 2.1 ± 0.1 c 2.7 ± 0.1 cd
12 0.3 ± 0.0 ab 0.7 ± 0.1 ab 2.8 ± 0.1 d 2.9 ± 0.1 d 4.5 ± 0.4 e
15 0.3 ± 0.0 ab 0.8 ± 0.1 b 3.1 ± 0.1 d 4.0 ± 0.2 e 6.1 ± 0.2 f
0 20 0.6 ± 0.0 ab 0.9 ± 0.1 b 4.4 ± 0.1 e 11.8 ± 0.1 i 23.0 ± 1.6 j
25 0.7 ± 0.1 ab 9.2 ± 0.3 g 9.8 ± 0.1 h – –
0 5 10 15 20 25 30 35 40 30 6.5 ± 0.1 f – – – –
35 11.9 ± 0.1 i – – – –
Baking time (min)
The furfural content was mean ± standard deviation (n = 3);
Fig. 4. Changes of moisture content in the sponge cake Model 11 during baking. Values with different letters are significantly different (p b 0.05).
 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445
Fig. 5. Concentration of HMF (mg kg−1) (in its logarithm form) in the sponge cake Model 11 during baking at 180, 190, 200, 210, 220 °C. (
kinetic at 220 °C. ( , y = 0.4822x−3.0154 R2 = 0.9974), kinetic at 210 °C. ( , y= 0.4107x−3.2740 R2 = 0.9914), kinetic at 200 °C. (
netic at 190 °C. ( , y = 0.3089x−4.4915 R2 = 0.9474), kinetic at 180 °C.
a drastic increase of the formation of HMF and F during the baking
process of sponge cakes. The concentrations of HMF and F were in
the range of 9.2–61.2 mg kg −1 (Fig. 2a) and 0.1–0.5 mg kg −1 in
sponge cakes in which a portion of the sucrose (80% out of 120%)
was replaced with other sugars. The amounts of HMF formed in
sponge cakes could be greatly influenced by the decomposition de-
gree of sucrose during baking, since reducing sugars were initially
lacking in batter. Results showed that replacing a portion of sucrose
with reducing sugars (e.g. glucose, fructose, lactose and maltose) pro-
moted formation of HMF in sponge cakes (Fig. 2a). While HMF was
formed at low concentrations in Model 1 cake (Fig. 2a), the decompo-
sition of sucrose into fructose and glucose in our cake model systems
may have been limited during baking at 205 °C for 11 min (Gökmen
et al., 2007).
Organic acids, such as citric acid, are sometimes used as the
acidulant to trigger CO2 production in leavening agents (Amrein,
Schönbächler, Escher, & Amado, 2004). In order to investigate the
effect of citric acid added in sponge cake batter on HMF and F forma-
tions, sponge cake batter was prepared by adding varying amounts of
citric acid and baked at 205 °C for 11 min. As shown in Fig. 2c and d,
adding citric acid (0 to 6%) to the sponge cake batter (Models 6–10),
lowering the pH from 7.5 to 3.4, resulted in a 442-fold increase of
HMF concentration and a 77-fold increase of F concentration. When
more citric acid (>4%) was added in Model 6, the batter reached a
pH lower than 4.0, and the rate of HMF and F formation increased.
This is most probably because the decomposition of sucrose into glu-
cose and fructose is promoted at pH lower than 4.0 (Pinheiro Torres,
Oliveira, Silva, & Fortuna, 1994). Lowering pH also increased the for-
mation of HMF in cookies (Gökmen et al., 2007).
3.2. Effect of baking conditions
In order to investigate the effects of baking time and temperature
on the change of pH, moisture content and the formation of HMF and
F, sponge cake batter (Models 11–15) was baked at different temper-
atures for different times. Changes in the texture and color of sponge
cake were the limiting factors in deciding the extent of baking
(Gökmen et al., 2007). Baking for less than 10 min at 180, 190 or
200 °C, and 8 min at 210 or 220 °C were found to be insufficient in
terms of sponge cake structure. Excessive surface browning of sponge
443
, y = 0.4572x−1.5787 R2 = 0.9371),
, y= 0.3227x−2.5820 R2 = 0.9945), ki-
cakes was prevented by limiting the baking times to 35 min at 180 °C,
to 25 min at 190 and 200 °C, and to 20 min at 210 and 220 °C.
With the increase of baking temperature and time, the pH value of
sponge cake Model 11 decreased as shown in Fig. 3. All five sponge
cake models (Models 11–15) followed similar trends. The pH value
of sponge cake models (Models 11–15) decreased from 7.5 ± 0.3 to
4.7 ± 0.1 during baking. The pH changes of sponge cakes would help
us understand the chemical changes occurring in sponge cakes during
baking. Caramelization causes the release of H +, thus the pH of the
sample undergoing caramelization falls with time, eventually into
the slightly sour region of pH 4–5 (Kroh, 1994).
Fig. 4 shows the changes in moisture content of the sponge cake
Model 11 during baking at different temperatures and times. The ini-
tial moisture content of the sponge cake model batter was 22.5 ±
0.4%, but there was a rapid decrease of the moisture content in
sponge cake models during the first 20 min of baking and reached a
moisture content of 3.1 ± 0.2% or lower within this period. All five
sponge cake models (Models 11–15) followed similar trends.
The F concentration in sponge cake models (Models 11–15) at
baking different temperature and time is listed in Table 2. With in-
creasing baking temperature and time, the formation of F increased.
Although F is formed during the Maillard reaction, it is principally
generated during caramelization, and HMF may be degraded to F
under high temperature conditions (Kroh, 1994). The F concentration
was in a range of 0.1–59 mg kg −1 (Model 12, glucose), and F concen-
tration below 1 mg kg −1 was found in 34% of the samples. The F con-
centration is far lower than HMF concentration in sponge cakes, and a
similar pattern was found in breakfast cereals (Rufián-Henares et al.,
2006), cookies (Ameur et al., 2006) and fortified wines (Pereira et al.,
2011).
The results depicted in Fig. 5 clearly show that with the increasing
of baking temperature and time, the concentration of HMF increased
in Model 11. All five sponge cake models (Models 11–15) followed
similar trends. The maximum concentration of HMF attained in the
sponge cake model was 4100±11 mg kg−1 (Model 13) and it occurred
when fructose was added and baked at baking 220 °C for 20 min. The
HMF levels found in sponge cakes are orders of magnitude higher than
what has been reported in cereal products: from 0.5 to 74.6 mg kg−1
in cookies (Ameur et al., 2006), from 6.6 to 240.5 mg kg−1 in breakfast
cereals (Rufián-Henares et al., 2006), from 11.46 to 32.06 mg kg−1 in in-
fant cereals (Fernández-Artigas, Guerra-Hernández, & García-Villanova,
444 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445

Table 3 the baking temperature, and was all the same for the five types of
Summary of kinetic analysis of HMF formation in five different sponge cake models. sugar. It is similar with the trend found in cookies during baking
T (°C) Regression equationa k R2 R2adj (Gökmen et al., 2007). One of the feasible pathways responsible for
this pattern of HMF formation could be the dehydration of hexose
Model 11
180 lnC(t) = 0.3089 t + ln0.0112 0.3089 0.9474 0.9369 sugars. The formation of one mole of HMF from one mole of hexose
190 lnC(t) = 0.3227 t + ln0.0756 0.3227 0.9945 0.9926 needs the release of three moles of water. In the early stages of the
200 lnC(t) = 0.4107 t + ln0.0379 0.4107 0.9914 0.9885 baking process, the presence of a high amount of water in the cake
210 lnC(t) = 0.4822 t + ln0.0490 0.4822 0.9974 0.9965
batter might inhibit or slow the reaction of sugar dehydration, once
220 lnC(t) = 0.4572 t + ln0.2062 0.4572 0.9371 0.9161
kf 108.7 the water evaporated the HMF shot out (Gökmen et al., 2007). The
Ea 22.1 monosaccharide yielded more content of HMF than disaccharide,
and the reducing sugar yielded more content of HMF than non-
Model 12 reducing sugar (Gökmen et al., 2007; Kroh, 1994), but it was not sig-
180 lnC(t) = 0.2030 t + ln1.1730 0.2030 0.9586 0.9503
nificantly different for the formation of F. It is reported that cookies
190 lnC(t) = 0.2043 t + ln3.9603 0.2043 0.9239 0.8985
200 lnC(t) = 0.2284 t + ln4.2236 0.2284 0.9008 0.8678 made of recipes lacking of reducing sugar had no or very limited
210 lnC(t) = 0.2675 t + ln3.9176 0.2675 0.9920 0.9893 quantity of HMF within 10 min of baking at 180 to 220 °C (Gökmen
220 lnC(t) = 0.3075 t + ln4.9361 0.3075 0.9979 0.9973 et al., 2008). The conversion of sucrose into glucose and fructose is
kf 41.9
necessary before the formation of HMF in sponge cakes. In this re-
Ea 20.3
spect, it is thought that any parameter, which affects the conversion
Model 13 of sucrose into glucose and fructose, would further affect the forma-
180 lnC(t) = 0.2700 t + ln0.2593 0.2700 0.9897 0.9877 tion of HMF in sponge cakes during baking. Overall results suggest
190 lnC(t) = 0.3016 t + ln0.5539 0.3016 0.9843 0.9791 that the level of HMF in cakes could be controlled by selecting
200 lnC(t) = 0.3477 t + ln0.6335 0.3477 0.9860 0.9814
lower baking temperature and longer baking times and avoiding the
210 lnC(t) = 0.3460 t + ln1.8379 0.3460 0.9772 0.9697
220 lnC(t) = 0.3806 t + ln2.9074 0.3806 0.8621 0.8162 use or the excessive use of reducing sugars in the recipe. In the
kf 16.3 sponge cake models of this work, baking at 180 °C for 25 min gave
Ea 15.4 a cake with lower levels of HMF and F.
HMF production from hexose is sensitive to the baking tempera-
Model 14
180 lnC(t) = 0.2233 t + ln0.3119 0.2233 0.9523 0.9428
ture and follows a first-order kinetics (Ameur et al., 2006; Ameur
190 lnC(t) = 0.3205 t + ln0.1705 0.3205 0.9879 0.9839 et al., 2007). The formation rate and concentration of HMF were en-
200 lnC(t) = 0.3136 t + ln0.4366 0.3136 0.9334 0.9112 hanced as the baking time and temperature increased as depicted in
210 lnC(t) = 0.3901 t + ln0.2289 0.3901 0.9830 0.9773 Table 3. These results indicate that the kinetics of HMF formation
220 lnC(t) = 0.4034 t + ln0.5839 0.4034 0.9721 0.9627
complied with Eq. (2) (first-order kinetics). The effect of fructose on
kf 228.4
Ea 25.8 HMF formation is much greater than the effects of sucrose, glucose,
lactose and maltose (Table 3). Fig. 6 shows the regression equation
Model 15 between the logarithmic relational expression with regard to the rate
180 lnC(t) = 0.2224 t + ln0.2452 0.2224 0.9781 0.9737
constant and the temperature of baking. The activation energy (Ea)
190 lnC(t) = 0.2895 t + ln0.3153 0.2895 0.9298 0.9064
200 lnC(t) = 0.2899 t + ln0.6593 0.2899 0.9933 0.9910
was calculated using the Arrhenius equation (k = kf exp(−Ea/RT)) and
210 lnC(t) = 0.3391 t + ln0.5640 0.3391 0.9819 0.9759 the regression equation in Fig. 6. The Ea of sponge cake models was in
220 lnC(t) = 0.3951 t + ln0.7603 0.3951 0.9519 0.9359 a range of 15.4–25.8 kJ mol−1. The Ea of Model 13 was the lowest, so
kf 146.6 the formation of HMF was more rapid than others. It could be confirmed
Ea 24.3
that fructose is more efficiently transformed in HMF than is glucose
Ea was calculated according to the Arrhenius equation k = kf exp(−Ea/RT). with the data obtained by Kroh (1994) in dessert wine and Ameur
T: the baking temperature (°C); C(t): the concentration of HMF (mg HMF per kg cake);
et al. (2007) in cookies, and that could be due to the fructose-specific re-
t: the baking time (min); k: rate constant (min−1); kf: the frequency factor (min−1);
Ea: the activation energy (kJ mol−1); R2: coefficient of determination; R2adj: adjusted R2. action pathway involving a 1,2-endiol intermediated in acidic media.
a
The HMF concentrations in the cake models were fitted to lnC(t)=kt+lnC0 for the The Ea of different foods are highly variable: 10.63 kJ mol−1 for cookies
first-order kinetics. (Ameur et al., 2006), 27.3 kJ mol−1 for apple cider (Gentry & Roberts,
2004), 43.01, 89.16 and 123.90 kJ mol−1 for three different types of
1999) and from 3.1 to 182.5 mg kg−1 in biscuits (Delgado-Andrade et al., rice wines (Chen et al., 2010), 32.5, 33.4, 33.7 and 43.6 kJ mol−1 for
2009). Ramírez-Jiménez, García-Villanova, and Guerra-Hernández
(2001) found that the toasting of bread increased HMF values from
-0.6
11.8 to 2024.8 mg kg−1 at baking from 0 to 60 min. This difference is
probably due to the higher baking temperature, longer baking time and Model 11
different sugar type in cakes during baking, which strongly favor HMF -0.9 Model 12
formation explaining the higher HMF concentration in the sponge cake
Model 13
models. The maximum concentration of HMF in sponge cake models is
ln k

Model 14
far more than 25 mg kg−1 regulated by EC Regulation No. 1493/99 for -1.2
grape must, but the risk assessment of HMF and limit content in food is Model 15
still a matter of debate and should be further evaluated. Various animal
-1.5
experiments reveal that no adverse effect levels are in the range of
80–100 mg kg−1 body weight and day (Abraham et al., 2011). So safety
margins are generally sufficient for the intake of HMF from sponge cakes. -1.8
The results depicted in Table 3 clearly show that both the intrinsic 0.002 0.0021 0.0022 0.0023
parameters (the type of sugar) and the extrinsic parameters (baking 1/T (K-1)
temperature and time) have strong influences on HMF formation
Fig. 6. Arrhenius plots for 5-hydroxymethylfurfural (HMF) accumulation. ( ), Model 11
in sponge cake models. An initial lower rate period was observed up y=-2661.5x+4.6886 R2 =0.8692. ( ), Model 12 y=-2442.5x+3.7355 R2 =0.9224.
to 15 min for sponge cake Models 11–15 baking at b 220 °C, which ( ), Model 13 y=-1849.2x+2.7934 R2 =0.9353. ( ), Model 14 y=
was followed by a rapid increase thereafter. This trend depended on -3101.2x+5.4311 R2 =0.8667. ( ), Model 15 y=-2924.2x+4.9876 R2 =0.9373.
 Y.-Y. Zhang et al. / Food Research International 49 (2012) 439–445
four types of unifloral honeys (Fallico et al., 2004). The Ea of different
food on HMF formation could be affected by some compounds (carbo-
hydrates, phenols or fats, etc.) (Chen et al., 2010), and water content
(Ameur et al., 2006). Lower water content and the presence of fructose
could favor HMF formation.
4. Conclusions
The effects of batter composition and baking conditions on the for-
mations of HMF and F were studied in sponge cakes. A fixed amount
of sucrose (40%) was included as minimum in sponge cake models to
improve the batter consistency and texture of sponge cakes. The
results showed that sugar type, baking temperature and time all
had strong influences on HMF and F formation in sponge cakes.
When the citric acid present in the batter of sponge cakes increases,
the conditions become more favorable for the formations of HMF
and F. Glucose and fructose yielded more HMF than sucrose, lactose
and maltose did in sponge cakes. With increasing baking temperature
and time, the pH value of the sponge cake decreased and the concen-
trations of HMF and F increased. HMF formation in sponge cake
models was determined as first-order kinetics. Fructose is more effi-
ciently transformed in HMF than are glucose and disaccharides. In-
creasing baking temperature, lowering water content and inclusion
of fructose all favored HMF formation. Lower baking temperatures,
longer baking times as well as the use of sucrose over simple sugars
is recommended to minimize HMF and F formation.
Acknowledgments
We are grateful for the financial support from the National Key
Technology R&D Program of China (no. 2011BAD23B01).
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