Food Research International 76 (2015) 661665

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Food Research International

journal homepage: www.elsevier.com/locate/foodres

Effects of pH on the formation of 4(5)-Methylimidazole in

glucose/ammonium sulfate and glucose/ammonium sulte
caramel model reactions
Xinlan Wu a, Fansheng Kong a, Minghui Huang a, Shujuan Yu a,b,c,
a
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China
b
State Key Laboratory of Pulp and paper Engineering, Guangzhou 510640, China
c
Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Guangzhou 510640, China

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

Article history: The objective of the present study was to detail the change of 4(5)-Methylimidazole (4-MI) in sulte and sulfate
Received 14 May 2015 reactions with different initial pH values. Glucose/ammonium sulfate and glucose/ammonium sulte reaction
Received in revised form 6 July 2015 systems with initial pH conditions 4.9, 5.9, 6.9, 8.0 and 8.6, were heated at 100 C for 2 h, respectively. Higher con-
Accepted 6 July 2015
centration of methylglyoxal (MGO) and 4-MI was detected in thermal treated glucose/ammonium sulte reac-
Available online 14 July 2015
tion system than that in sulfate system. The SO2 3 reacting with MGO and other precursors of 4-MI at higher
Keywords:
pH conditions prevented 4-MI formation. However, no inhibition of 4-MI was found at lower pH conditions
4(5)-Methylimidazole due to higher reactivity of the nucleophilic NH+ 2
4 than SO3 . The browning intensity of the sulte system changed

pH scarcely at higher pH values, which was possibly caused by the polyreaction between SO2 3 and carbonyl, instead
Caramel of the intermolecular polymerisation of carbonyl in the advanced stage of the Maillard reaction.
Maillard reaction 2015 Elsevier Ltd. All rights reserved.

Chemical compounds studied in this article:

4(5)-Methylimidazole (PubChem CID: 13195)
Methylglyoxal (PubChem CID: 880)
2-Methylquinoxaline (PubChem CID: 23686)

1. Introduction formed in the Maillard reaction, 4-MI was a concern to researchers

Caramel colours have been widely used in various food products as
colourants, including baked food, beers, spirits, wines, beverages, pickles,
sauces, jam, ice creams and confectionary (Seo, Ka, & Lee, 2014). There are
four types of caramel colours which are named class I, II, III and IV (Hengel
& Shibamoto, 2013; Seo et al., 2014; Yamaguchi & Masuda, 2011). Class I
caramel colour is produced by the thermal treatment of carbohydrates.
Class II caramel colour is prepared via the reaction between carbohy-
drates and sulte compounds. Class III caramel colour is produced by
heating carbohydrates and ammonium compounds. And class IV caramel
colour is prepared by reacting carbohydrates, ammonium compounds
and sulte compounds at a high temperature (Lee, Jang, & Shibamoto,
2013; Sengar & Sharma, 2014; Seo et al., 2014).
In the process of class III and IV caramel colours, a major reaction that
occurred was the Maillard reaction, where carbonyl compounds react
with amino groups or ammonia (Jang, Jiang, Hengel, & Shibamoto,
2013; Moretton, Cretier, Nigay, & Rocca, 2011). Among the products
Corresponding author at: College of Light Industry and Food Sciences, South China
University of Technology, Guangzhou 510640, China.
E-mail address: shujuanyu8@gmail.com (S. Yu).
because of its carcinogenic risk. The National Toxicology Program
(NTP) has identied 4-MI as a carcinogen in 2007 (National Toxicology
Program, 2007), and the Ofce of Environmental Health Hazard Assess-
ment (OEHHA) within the California Environmental Protection Agency
recommended that the acceptable daily intake (ADI) of 4-MI must be
lower than 29 g/day (Ofce of Environmental Health Assessment,
2011). In addition, 4-MI is classied as group 2B (possibly carcinogenic
in humans) by the International Agency for Research on Cancer (IARC)
Monographs (IARC, 2012).
4-MI was rstly reported to be formed in the Maillard reaction that
consists of D-glucose and ammonia in the early 1962 (Seo et al., 2014).
Later, many researchers have focused on the levels of 4-MI in food prod-
ucts (Casal, Fernandes, Oliveira, & Ferreira, 2002; Cunha, Barrado, Faria,
& Fernandes, 2011; Fernandes & Ferreira, 1997; Hengel & Shibamoto,
2013; Klejdus, Moravcova, & Kuban, 2003; Klejdus, Moravcova,
Lojkova, Vacek, & Kuban, 2006). For instance, 4-MI was detected in com-
mercial roasted coffee ranging from 0.307 to 1.231 mg/kg (Casal et al.,
2002). Several hypothetical formation pathways of 4-MI in the Maillard
reaction have been proposed in recent years (Jang et al., 2013; Moon &
Shibamoto, 2011). Based on the hypothesis, 4-MI originated from the
reaction of MGO, formaldehyde and ammonia. Many factors may

http://dx.doi.org/10.1016/j.foodres.2015.07.011
0963-9969/ 2015 Elsevier Ltd. All rights reserved.
662 X. Wu et al. / Food Research International 76 (2015) 661665
inuence the formation of 4-MI in caramel colour production. Report
has suggested sulphite was a signicant impact on the formation of
4-MI (Lee et al., 2013). However, another important factor, i.e. pH
value, was always ignored in recent literatures.
The objective of the present study was to investigate the role of pH
on 4-MI formation in caramel model system. Generally, the ammonium
compounds used in caramel colour production are hydroxides, carbon-
ates, bicarbonates, phosphates, sulfates, sultes and bisultes, and the
sulte compounds used are sulfurous acid and sultes and bisultes
of potassium, sodium, and ammonium (Sengar & Sharma, 2014).
Thus, glucose/ammonium sulfate and glucose/ammonium sulte were
chosen as the materials for preparing class III and class IV caramel
colours, respectively.
2. Materials and methods
2.1. Chemicals
D-glucose (99%), ammonium sulfate (99%), ammonium sulte
(99%), o-phenylenediamine (OPD, 99%), methylglyoxal (25%), 2-
methylquinoxaline (2-MQ, 99%), formaldehyde (25%), 4-MI (99%),
disodium hydrogen phosphate dodecahydrate (99%), disodium
dihydrogen phosphate dehydrate (99%) and potassium dihydrogen
phosphate dehydrate (99%) were purchased from Sigma-Aldrich
(St. Louis, MO, USA). Methanol was chromatographic grade and obtained
from Merck (Damstadt, Germany).
2.2. Heating procedure
A mixture of D-glucose (4 mmol) and ammonium sulfate (2 mmol),
or D-glucose (4 mmol) and ammonium sulte (2 mmol), was dissolved
in 4 mL of different buffers (1 M K2HPO4/NaH2PO4, pH 4.9, 5.9 6.9 7.9
and 8.6), respectively. The mixtures were heated in screw-cap tubes
using an oil bath at 100 C for 2 h. At predetermined heating time, sam-
ples were taken and immediately cooled in iced water. 200 L sample
was mixed with 800 L o-phenylenediamine solution (1 mg/mL in
1 M phosphate buffer solution (PBS) of pH 7.4), and then incubated at
room temperature for 12 h. The remainder of the sample was stored
at 4 C for further analyses.
A mixture of MGO (0.4 mmol), formaldehyde (0.4 mmol) with
ammonium sulfate (0.4 mmol) or ammonium sulte (0.4 mmol) was
operated in the same procedure as stated above, except the heating
time was set at 30 min. All samples were stored at 4 C until used.
2.3. HPLC-UV analysis of quinoxaline formed from MGO with OPD
MGO was measured using the corresponding quinoxalines 2-MQ
(Fig. 1). After the derivatisation with OPD, samples were ltered
through 0.45 L Millex-HNnylon lters (Millipore, Billerica, USA). Then
the percolates were analysed by a reverse-phase high-performance liq-
uid chromatography (RP-HPLC) system, which consisted of a Waters
600 pump, a Rheodyne 7725i manual injector, Waters XBridge BEH
Shield RP18 Column (5 m, 4.6 mm 250 mm) and a Waters 2998
Fig. 1. Chemical structure of corresponding quinoxaline of MGO obtained via
derivatisation with OPD.
DAD (Waters Corp., Milford, USA). The injection volume was 20 L
and the mobile phase (watermethanol system) was as follows: a linear
gradient of 5% methanol to 20% methanol, from 0 to10 min, then a linear
gradient of 20% methanol to 30% methanol, from 10 to 28 min, and a
linear gradient of 30% methanol to 100% methanol, from 28 to 36 min,
and at last, 100% methanol from 36 to 45 min. The ow rate was set
at 1.0 mL/min, and the detection wavelength was set at 315 nm. The
column temperature was 25 C.
2.4. Measurement of browning intensity
Browning intensity of glucose/ammonium sulfate and glucose/
ammonium sulte reacted solutions was measured according to
literature with a slight modication (Guan et al., 2011). 100-fold dilu-
tion of the treated solution was made using distilled water and the ab-
sorbance was measured at 420 nm using a UV-1810 spectrophotometer
(Puxi, Beijing, China).
2.5. Analysis of 4-MI
Glucose/ammonium sulfate and glucose/ammonium sulte reacted
solutions were pretreated according to the method described by the
Moon with a minor modication (Moon & Shibamoto, 2011). Briey,
3 mL reacted solution was passed through an Ansys SPEC SCX Disc
15 mg/3 mL cartridge (Varian, Walnut Creek, CA) after the addition of
3 mL of 0.1 N HCl, and then the cartridge was washed with 3 mL meth-
anol. The retained compounds in the cartridge were eluted out with
6 mL of methanol/ammonia (5%, v/v). The collected extracts were
dried using a rotary evaporator at 40 C, then the residues were dis-
solved with 3 mL deionised water for 4-MI analysis. The HPLC-MS/MS
system was used to detect 4-MI, which was referred to the literature
with a slight modication (Moon & Shibamoto, 2011). The HPLC-MS/MS
system consisted of a Waters HPLC system (Waters Co., Milford, USA),
coupled with a Thermo Scientic (San Jose, USA) LTQ mass spectrometer.
Chromatographic separation was accomplished with a Waters XBridge
BEH Shield RP18 Column (3.5 m, 3.0 mm 150 mm) using mobile
phase A (10 mM of ammonia in high-purity water) and mobile phase B
(methanol) in a gradient programme with a ow of 0.3 mL/min:
010 min, 5% B; 1115 min, 5% B to 100% B; 1520 min, 100% B. Mass
spectrometry was performed via an electrospray ionisation (ESI) source
operated in the positive ion mode at 275 C with nitrogen gas (35 psi),
and the collision energy was set at 33 V. The mass spectrometer was op-
erated in selected reaction monitoring (SRM) mode, and the 4-MI peak
area with the transition of m/z 83 to 56 was used for the conrmation
and quantication.
A standard calibration curve was obtained in 0.2, 0.5, 1.0, 2.0, 5.0
and 10.0 g/mL concentrations by diluting the stock 4-MI solution
(1 mg/mL) using distilled water. The regression equation for 4-MI
standard was as in Eq. (1)
 
y 21141x3174:6 R2 0:9953 1
where x is the concentration of 4-MI, g/mL; and y is the peak area of
4-MI.
2.6. Statistical analyses
All experiments were carried out at least in triplicate. Means and
standard errors of data were calculated for each treatment. Analysis of
variance (ANOVA) was carried out using SAS 9.3 software (SAS Institute
Inc., Cary, NC) to determine any signicant differences between treat-
ments (P b 0.05).
 X. Wu et al. / Food Research International 76 (2015) 661665
3. Results and discussion
3.1. MGO formed in caramel model systems
Researchers reported that MGO was the main precursor of 4-MeI
(Hengel & Shibamoto, 2013; Moon & Shibamoto, 2011). MGO should
undergo derivatisation into quinoxaline adduct (Fig. 1) before HPLC
analyses (Degen, Hellwig, & Henle, 2012; Mavric, Wittmann, Barth, &
Henle, 2008; Wang, Yagiz, Buran, Nunes, & Gu, 2011). In this study,
the sample was derived using OPD before HPLC analysis, and Fig. 2
presented the content of MGO formed in glucose/ammonium sulfate
and glucose/ammonium sulte caramel model systems. Obviously, an
increase in pH of the above caramel model systems led to the increase
of MGO. Additionally, the increase of MGO in the Maillard reaction
was probably due to the stronger nucleophilicity of NH+ 4 at higher pH
values. This result was in agreement with Martins et al. who found
that more MGO was formed in a higher pH condition (Martins & Van
Boekel, 2005).
On the other hand, more MGO was formed in the glucoseammonium
sulte system than in the glucoseammonium sulfate reaction system
at all selected pH values, which indicated that sulte can accelerate the
formation of MGO. This result was in accordance with Yu et al. who
found that the addition of sulte increased the content of MGO in the deg-
radation of 2-Amino-2-Deoxyglucose (Yu, Ma, & Xu, 2013). It indicated
that the increase of MGO in glucoseammonium sulte reaction system
is mainly due to the sulte-induced decomposition of the Schiff base
(ie. 2-Amino-2-Deoxyglucose) into MGO. Wang and Ho (2012) reported
that MGO can be produced through non-oxidative reactions of the Schiff
base. Obviously, the reducing sulte can inhibit the oxidative reactions of
the Schiff base. On the contrary, the non-oxidative reactions of the Schiff
base might be accelerated. As a result, MGO was promoted by sulte.
However, more experiments are needed to prove this hypothesis.
3.2. 4-MI analysis
Fig. 3 showed the results of 4-MI content in the reacted solutions. In
the glucose/ammonium sulfate caramel model system, the content of
4-MI increased as pH increased. When pH increased from 4.9 to 8.6,
the content of 4-MI changed from 3.13 to 37.09 M/L. The change of
4-MI was in accordance with that of MGO. It is a good correlation
(R2 = 0.973) between MGO and 4-MI. This result indicated that the
increased 4-MI was mainly due to the increase of MGO in the glucose
ammonium sulfate caramel model system. However, the change of
4-MI in the glucose/ammonium sulte system was not the same
as that in the glucose/ammonium sulfate caramel model system.
Increasing pH from 4.9 to 6.9 accelerated the formation of 4-MI (varied
Fig. 2. Content of MGO formed in glucose/ammonium sulfate and glucose/ammonium
sulte caramel model systems. Bars with the different letters (or the different labelled
(') letters) are signicantly different (P b 0.05).
663
Fig. 3. Content of 4-MI formed in glucose/ammonium sulfate and glucose/ammonium
sulte model systems. Bars with the different letters (or the different labelled (') letters)
are signicantly different (P b 0.05).
from 38.32 to 97.94 M/L). However, a signicant decrease of 4-MI was
observed when the pH increased from 6.9 to 8.0 and 8.6 (changed from
97.94 M/L to 73.39 M/L and 69.14 M/L), respectively. This phenome-
non suggested that 4-MI could be reduced at a higher pH value in the
presence of sulte. Lee et al. (2013) have found that sulte can inhibit
the formation of 4-MI at high concentration. Base on the results, Jang
et al. (2013) proposed a formation pathway of 4-MI in the presence of
sulte. The proposed pathway is important to our understanding on
the changes of 4-MI at different pH values.
In order to further study the mechanism of pH inuencing the
formation of 4-MI, two model reactions of MGO/formaldehyde/
ammonium sulfate and MGO/formaldehyde/ammonium sulte were
investigated, respectively. In the MGO/formaldehyde/ammonium
sulfate model system, the higher content of 4-MI was detected at higher
initial pH (Fig. 4), which indicated that higher pH could promote the
formation of 4-MI. This may be a second reason that a higher content
of 4-MI was obtained when the pH was increased in the glucose/
ammonium sulfate caramel model system. In the MGO/formaldehyde/
ammonium sulte model system, adjusting the pH from 4.9 to 6.9
increased the quantity of 4-MI. However, adjusting the pH from 6.9 to
8.6 reduced the 4-MI content signicantly (Fig. 4). This result was in ac-
cordance with the change of 4-MI in the glucoseammonium sulte car-
amel model system. The possible reason that explains these phenomena
was detailed as follows. Fig. 5 presented the proposed formation path-
way of 4-MI in the absence of sulte (Jang et al., 2013; Seo et al.,
2014; Yaylayan & Haffenden, 2003). 4-MI was formed by the reaction
of MGO, formaldehyde and NH+ 4 through pathway B. Additional SO3
2
Fig. 4. Content of 4-MI formed in MGO/formaldehyde/ammonium sulfate and
MGO/formaldehyde/ammonium sulte model systems. Bars with the different letters
(or the different labelled (') letters) are signicantly different (P b 0.05).
664 X. Wu et al. / Food Research International 76 (2015) 661665
Fig. 5. Proposed formation pathway of 4-MI in the presence of SO2
3 , adapted from Jang et al. (2013), Seo et al. (2014) and Yaylayan and Haffenden (2003).
competitively reacted with NH+ 4 to prevent forming the precursor of
4-MI at higher pH conditions (paths A and C). At lower pH values,
NH+ 4 reacted more easily with carbonyl compounds comparing with
SO2 + 2
3 , caused by higher reactivity of the nucleophilic NH4 than SO3 .
Thus, the main reaction at lower pH was through pathway B to form
4-MI. However, as the pH was increased to 8.0 and/or 8.6, the SO23
held higher reactivity with carbonyl compounds, resulting in more
sulte-aldehyde adducts that formed through pathway A and C, and
inhibited the formation of 4-MI. Compared with the two caramel
systems, the difference of the change of 4-MI was mainly caused by
the nucleophilic and reduced SO23 .
3.3. Changes of browning intensity
The wavelength used in the present study ( = 420 nm) has been
widely used for evaluating the colour intensity of the Maillard reaction
products (Lee et al., 2013). The formation of brown pigments was due
to the polymerisation of the intermediates (dicarbonyl compounds,
aldehydes, etc.) in the advanced stages of the Maillard reaction (Guan,
Fig. 6. Changes of Abs420 of glucose/ammonium sulfate and glucose/ammonium sulte
caramel model systems. Bars with the different letters (or the different labelled (') letters)
are signicantly different (P b 0.05).
Zhu, Yu, Xu, & Shi, 2011; Lertittikul, Benjakul, & Tanaka, 2007). Fig. 6
showed the absorbance at 420 nm (Abs420) of the solution. In the
glucose/ammonium sulfate caramel model system, a signicant in-
crease of Abs420 was observed as pH increased. It shows a good correla-
tion (R = 0.975) between MGO and the Abs420. This result was probably
caused by the rapid formation of active carbonyl compounds at high pH
conditions. Consequently, more brown polymers were formed and
higher Abs420 were detected. In the glucose/ammonium sulte caramel
model system, Abs420 absorbance increased signicantly when pH
changed from 4.9 to 6.9. But Abs420 tested in the model systems
changed slightly when pH increased from 6.9 to 8.6. A possible explana-
tion could be that the SO2
3 reacted with more carbonyl compounds due
to the increase of its nucleophilicity at higher pH conditions, and limited
the polymerisation of carbonyl compounds.
4. Conclusion
Higher content of MGO, i.e. 4-MI precursor compound, was detected
in the glucose/ammonium sulte system, comparing with that in the
glucose/ammonium sulfate system. Increased pH can decrease the con-
tent of 4-MI in the presence of sulte due to that SO23 reacted with
MGO and other precursors of 4-MI. However, NH+ 4 reacted more easily
with carbonyl compounds at lower pH conditions, caused by higher re-
activity of the nucleophilic NH+ 2
4 than SO3 , leading to more 4-MI for-
mation at pH 4.9 to 6.9. The browning intensity of the glucose/
ammonium sulte reaction system was inhibited at high pH possibly
caused by the reaction between SO23 and carbonyl compounds,
which limited the polymerisation of carbonyl compounds in the ad-
vanced stage of the Maillard reaction.
Acknowledgements
All authors acknowledge the National Science Foundation of
China (Grant No.31271889 and Grant No.31071564), and the
Science and Technology Planning Project of Guangdong Province,
China (No.2014B020205001 and No.2013J4500036).
 X. Wu et al. / Food Research International 76 (2015) 661665
References
Casal, S., Fernandes, J.O., Oliveira, M., & Ferreira, M.A. (2002). Gas chromatographicmass
spectrometric quantication of 4-(5-)-methylimidazole in roasted coffee after ion-
pair extraction. Journal of Chromatography A, 976(12), 285291.
Cunha, S.C., Barrado, A.I., Faria, M.A., & Fernandes, J.O. (2011). Assessment of 4-(5-)
methylimidazole in soft drinks and dark beer. Journal of Food Composition and Analysis,
24(45), 609614.
Degen, J., Hellwig, M., & Henle, T. (2012). 1,2-Dicarbonyl compounds in commonly
consumed foods. Journal of Agricultural and Food Chemistry, 60(28), 70717079.
Fernandes, J.O., & Ferreira, M.A. (1997). Gas chromatographic mass spectrometric
determination of 4-(5) methylimidazole in ammonia caramel colour using ion-pair
extraction and derivatization with isobutylchloroformate. Journal of Chromatography
A, 786(2), 299308.
Guan, Y.G., Yu, P., Yu, S.J., Xu, X.B., Shi, W.H., & Sun, W.W. (2011). Effects of pressure on
the glucoseammonium sulphite caramel solutions. Food Chemistry, 127(2), 596601.
Guan, Y.G., Zhu, S.M., Yu, S.J., Xu, X.B., & Shi, W.H. (2011). SO(3)2-effects the
5-hydroxymethyl-2-furaldehyde content in ammonium sulphite-glucose solutions.
International Journal of Food Science and Technology, 46(5), 10071013.
Hengel, M., & Shibamoto, T. (2013). Carcinogenic 4 (5)-methylimidazole found in
beverages, sauces, and caramel colours: Chemical properties, analysis, and biological
activities. Journal of Agricultural and Food Chemistry, 61(4), 780789.
IARC monographs ()). 4-Methylimidazole. URL Monographs.iarc.fr/ENG/Monographs/
vol101/mono101-015.pdf (accessed Oct 16, 2012)
Jang, H.W., Jiang, Y.P., Hengel, M., & Shibamoto, T. (2013). Formation of 4(5)-
methylimidazole and its precursors, alpha-dicarbonyl compounds, in Maillard
model systems. Journal of Agricultural and Food Chemistry, 61(28), 68656872.
Klejdus, B., Moravcova, J., & Kuban, V. (2003). Reversed-phase high-performance liquid
chromatographic/mass spectrometric method for separation of 4-methylimidazole
and 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole at pg levels. Analytica Chimica
Acta, 477(1), 4958.
Klejdus, B., Moravcova, J., Lojkova, L., Vacek, J., & Kuban, V. (2006). Solid-phase extraction
of 4(5)-methylimidazole (4Mel) and 2-acetyl-4(5)- (1,2,3,4-tetrahydroxybutyl)-
imidazole(THI) from foods and beverages with subsequent liquid chromatographic-
electrospray mass spectrometric quantication. Journal of Separation Science, 29(3),
378384.
Lee, K.G., Jang, H., & Shibamoto, T. (2013). Formation of carcinogenic 4(5)-methylimidazole
in caramel model systems: A role of sulphite. Food Chemistry, 136(34), 11651168.
Lertittikul, W., Benjakul, S., & Tanaka, M. (2007). Characteristics and antioxidative activity
of Maillard reaction products from a porcine plasma proteinglucose model system
as inuenced by pH. Food Chemistry, 100(2), 669677.
665
Martins, S.I., & Van Boekel, M.A. (2005). Kinetics of the glucose/glycine Maillard reaction
pathways: Inuences of pH and reactant initial concentrations. Food Chemistry, 92(3),
437448.
Mavric, E., Wittmann, S., Barth, G., & Henle, T. (2008). Identication and quantication of
methylglyoxal as the dominant antibacterial constituent of Manuka (Leptospermum
scoparium) honeys from New Zealand. Molecular Nutrition & Food Research, 52(4),
483489.
Moon, J.K., & Shibamoto, T. (2011). Formation of carcinogenic 4(5)-methylimidazole
in Maillard reaction systems. Journal of Agricultural and Food Chemistry, 59(2),
615618.
Moretton, C., Cretier, G., Nigay, H., & Rocca, J.L. (2011). Quantication of 4-methylimidazole
in class III and IV caramel colours: Validation of a new method based on heart-cutting
two-dimensional liquid chromatography (LC-LC). Journal of Agricultural and Food
Chemistry, 59(8), 35443550.
National Toxicology Program (2007). Toxicology and carcinogenesis studies of
4-methylimidazole (CAS No. 822-36-6) in F344/N rats and B6C3F1 mice (Feed Studies).
NTP Technical Report Series No. 535, NIH Publication No. 07-4471. Research
Triangle Park, NC: U.S. Department of Health and Human Service.
Ofce of Environmental Health Assessment (2011). Notice of intent to list
4-methylimidazole. URL http://oehha.ca.gov/prop65/crnr_notices/admin_listing/
intent_to_list/noilpkg32.html (Accessed 14.01.18)
Sengar, G., & Sharma, H.K. (2014). Food caramels: A review. Journal of Food Science and
Technology-Mysore, 51(9), 16861696.
Seo, S., Ka, M. -H., & Lee, K. -G. (2014). Reduction of carcinogenic 4 (5)-methylimidazole
in a caramel model system: Inuence of food additives. Journal of Agricultural and
Food Chemistry, 62(27), 64816486.
Wang, Y., & Ho, C. -T. (2012). Flavour chemistry of methylglyoxal and glyoxal. Chemical
Society Reviews, 41(11), 41404149.
Wang, W., Yagiz, Y., Buran, T.J., Nunes, C.D., & Gu, L.W. (2011). Phytochemicals from
berries and grapes inhibited the formation of advanced glycation end-products by
scavenging reactive carbonyls. Food Research International, 44(9), 26662673.
Yamaguchi, H., & Masuda, T. (2011). Determination of 4 (5)-methylimidazole in soy sauce
and other foods by LC-MS/MS after solid-phase extraction. Journal of Agricultural and
Food Chemistry, 59(18), 97709775.
Yaylayan, V.A., & Haffenden, L.J.W. (2003). Mechanism of imidazole and oxazole
formation in C-13-2-labelled glycine and alanine model systems. Food Chemistry,
81(3), 403409.
Yu, S., Ma, F., & Xu, X. (2013). Effects of sulte on degradation of 2-amino-2-deoxyglucose
in Maillard reaction. Journal of South China University of Technology (Natural Science
Edition), 10, 007.