Food Sci. Technol. Res.

, 18 (1), 67 – 76, 2012

Determination of Furosine and Fluorescence as Markers of the Maillard Reaction for

the Evaluation of Meat Products during Actual Cooking Conditions

Keiko Yamaguchi1*, Yuri Nomi2, Takeshi Homma1, Midori Kasai2 and Yuzuru Otsuka2

1
Department of Food and Nutrition, Japan Women’s University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan
2
Graduate School of Humanities and Sciences, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610, Japan

Received August 9, 2011; Accepted September 5, 2011

During food processing and cooking, various flavor compounds, colored substances and toxic sub-
stances are produced by the Maillard reaction. It is important to investigate the relationship between the
Maillard reaction products and the actual cooking conditions. Therefore, we investigated the effects of
seasonings and reaction conditions on the Maillard reaction of meat using furosine and fluorescent com-
pounds. The addition of 1.0% NaCl decreased the reaction rate of the Maillard reaction measured by
furosine in Glc - meat reaction. However, the reaction rate of meat with added sucrose did not change.
Furosine content in meat with added sucrose increased rapidly when it was pan-broiled and fried, but in-
creased slowly at the beginning when it was baked in a gas oven. The fluorescence of broiled meat showed
a maximum at 333/425 nm (exc./em.) and is a useful indicator of the Maillard reaction of meat samples.
The rate of increasing fluorescence in baked meat was lower than that of meat heated by other cooking
methods.

Keywords: maillard reaction, lysine, meat, browning, furosine, fluorescence

Introduction thermore, many studies indicated that the Maillard reaction

During food processing, storage, and cooking, the reac-
tion between the amino group and the carbonyl group of
food components occurs randomly. This reaction is called the
amino-carbonyl reaction, or the Maillard reaction. The pre-
dominant early glycation adduct is fructosyl-lysine, which
degrades slowly to form many different advanced glycation
end products (AGEs).
It is now known that various substances are generated in
food processing and cooking by the Maillard reaction, such
as flavor compounds, colored substances and toxic substanc-
es. The major flavor compounds produced by the Maillard
reaction are heterocyclic compounds: thiophenes, thiazoles,
pyrazines, pyrroles, imidazoles, and pyridines. Melanoidins
are polymeric brown compounds formed in the last stage of
the Maillard reaction. Flavor and color are important fac-
tors in consumer acceptance because they are related to food
quality and characteristics. On the other hand, heterocyclic
amines (HCAs) and acrylamide are toxic substances. Fur-
*To whom correspondence should be addressed.
E-mail: yama-k@fc.jwu.ac.jp
produced imidazoles including 4(5)-methylimidazole (Shib-
amoto, 1983). The carcinogenic 4(5)-methylimidazole exists
generally in foods and beverages (Moon and Shibamoto,
2011). Hence, it is important to investigate the mechanism of
the Maillard reaction during the actual cooking conditions to
improve food quality and safety.
Fructosyl-lysine, an Amadori compound, is formed in
the early stage of the Maillard reaction, and is measured
by furosine, which is generated from the acid hydrolysis of
fructosyl-lysine. Therefore, the determination of furosine is
a useful marker of the early stage of the Maillard reaction.
Heyns et al. (1968) and Finot et al. (1968) identified the
structure of furosine simultaneously as ε-N-(2 furoylmethyl)-
L-lysine. The use of furosine was further examined using
improved high performance liquid chromatography (HPLC)
techniques (Chiang, 1983; Resmini et al., 1990; Tirelli and
Pellegrino, 1995). Analyses of furosine in heat-treated foods
have increased since 1992 and have been applied worldwide.
Furosine content in dairy products has attracted attention,
because the Maillard reaction between lactose and milk pro-
tein is responsible for the loss of available lysine during food
68
processing and storage (Bosch et al., 2008; Erbersdobler et
al., 1987; Tokusoglu et al., 2006; Villamiel et al., 2000). Fu-
rosine analyses have been applied to milk products, cereals,
pasta, honey, and other items (Delgado-Andrade et al., 2007;
Erbersdobler and Hupe, 1991; Garcia-Banos et al., 2004;
Sanz et al., 2003). However, meat as the main source of
protein has not been well analyzed. Pompei and Spagnolello
(1997) were the only researchers to analyze the formation
of Amadori products by furosine in seasoned and processed
pork compared to that in raw meat. They reported that the
furosine content in tumbled meat containing NaCl and Glc
in the temperature range of 70 − 90℃ increased more signifi-
cantly than that in raw meat. Since the common sugar used
as a seasoning, sucrose, is a non-reducing sugar, it is thought
that the contribution of sucrose to the formation of Amadori
products is rather small. Although actual cooking involves
various heat settings, only several studies have examined
Amadori products with furosine.
There are several methods to detect AGEs. Ahmed and
Thornalley (2007) reported that the best method to detect
Maillard reaction products is the LC/MS/MS method. How-
ever, this method has not yet been established, and AGE
immunoassay procedures are associated with significant
problems and limitations. The “total AGE fluorescence” with
excitation and emission wavelengths of 350 and 450 nm,
respectively, is used to investigate AGE formation in vivo.
Some AGEs have fluorescent properties, particularly pentosi-
dine, which shows fluorescence at an excitation wavelength
of 335 nm and an emission wavelength of 385 nm (Sell and
Monnier, 1989; Gerrard, 2002). Pentosidine was found in
only very small amounts in foods (Henle et al., 1997). AGEs
generated in vivo are considered to be related to aging and
diabetes (Brownlee, 2001). AGEs from food might have the
same effect once they are ingested into the body (Hofmann
et al., 2002; Uribarri et al., 2005; Vlassara et al., 2002).
However, the effect of cooking methods on AGE formation
in meat has not been previously measured by fluorescence.
The aim of this paper is to investigate the relationship
between Maillard reaction products and actual cooking con-
Table 1. Composition of the Maillard reaction samples with L-Lys, saccharide and NaCl.
Model name L-Lys (M)
Lys 0.10
Lys-NaCl 0.10
Lys-Glc 0.10
Lys-Sucrose 0.10
Lys-Glc-NaCl 0.10
Lys-Sucrose-NaCl 0.10
K. Yamaguchi et al.
ditions by analysis of browning compounds, furosine and
fluorescent compounds in order to identify better cooking
conditions for reducing AGEs.
Materials and Methods
Materials L-Lys, D-Glc, and NaCl were purchased
from Kanto Chemical (Tokyo, Japan). Sucrose was pur-
chased from Wako Pure Chemical Industries (Osaka, Japan).
Furosine was purchased from Neosystem Laboratoire (Stras-
bourg, France). All the other reagents were of analytical
grade. Ground beef (thigh meat) of Japanese origin was used.
Preparation of Maillard reaction samples containing L-
Lys, saccharide (D-Glc or Sucrose) and NaCl L-Lys, sac-
charide (D-Glc or Sucrose) and NaCl were dissolved in 0.2
M phosphate buffer (pH 6.0) as shown in Table 1. Test tubes
(16 × 100 mm) containing 8.0 mL of mixed solution were
sealed and heated in a block heater at 100℃ for 0 − 120 min.
After heating, the tubes were immediately cooled and stored
in a refrigerator.
Preparation of pan-broiled meat samples containing sac-
charide and NaCl Meat, saccharide (D-Glc or Sucrose) and
NaCl were mixed according to Table 2, and 50 g of mixed
sample was made into a hamburger steak shape. The samples
were heated at 200℃ for 0 − 62 min in a Teflon pan with no
oil, and broiled on both sides. The surface temperature of the
pan was measured by a Minolta spot thermometer (HT-21,
Minolta Co., Ltd., Osaka, Japan) and kept at 200℃. After
heating, the samples were immediately cooled and homoge-
nized with ultra pure water by a homogenizer (Nippon Seiki,
Tokyo, Japan). The paste samples were freeze-dried and
stored in a refrigerator.
Preparation of meat samples containing white superior
soft sugar and NaCl, heated using three types of cooking
methods for furosine analysis Meat was mixed with 5.0%
white superior soft sugar and 1.0% NaCl, and 50 g of mixed
sample was made into a hamburger steak shape as seasoned
meat. The concentration of seasoning was based on the typi-
cal concentration used in homemade foods. In addition, 50
g of raw meat sample was also made into a hamburger steak
D-Glc(M) Sucrose (M) NaCl (%)
0.00 0.00 0.00
0.00 0.00 1.00
0.15 0.00 0.00
0.00 0.15 0.00
0.15 0.00 1.00
0.00 0.15 1.00
Markers of the Maillard Reaction of Meat
Table 2. Composition of pan-broiled meat samples containing saccharide and NaCl.
Model name D-Glc (%)
Meat 0.0
Meat-NaCl 0.0
Meat-Glc 5.0% 5.0
Meat-Glc 2.5% 2.5
Meat-Sucrose 10.0% 0.0
Meat-Sucrose 5.0% 0.0
Meat-Nacl-Glc 5.0% 5.0
Meat-NacCl-Glc 2.5% 2.5
Meat-NaCl-Sucrose 10.0% 0.0
Meat-NaCl-Sucrose 5.0% 0.0
shape as non-seasoned meat. The samples were pan-broiled
(at 200℃), baked using a gas oven (at 170℃) or fried (at
140℃) for 0 − 30 min. The baked meat samples were placed
at the center of the oven (300 × 425 × 375 mm, GMO-S1300,
Harman Co. Ltd., Osaka, Japan). Fried meat samples were
fried with 100% pure canola oil. The surface temperature
of the oil was measured by a Minolta spot thermometer HT-
21 and kept at 140℃. The internal temperature of each meat
sample was measured by a data collector (AM-7002, Anritsu
Meter Co. Ltd., Tokyo, Japan) with thermocouple probe type
K (Anritsu Meter Co. Ltd., Tokyo, Japan). After heating, the
meat samples were immediately cooled and homogenized
with ultra pure water using a homogenizer. The paste sam-
ples were freeze-dried and stored in a refrigerator.
Preparation of meat samples heated using three types of
cooking methods for fluorescence analysis Fifty grams of
raw meat sample was made into a hamburger steak shape.
The meat samples were pan-broiled (at 200℃), baked in a
gas oven (at 170℃) and fried (at 140℃) for 0 − 30 min, as
described above. After heating, the meat samples were im-
mediately cooled and homogenized with ultra pure water us-
ing a homogenizer. The paste samples were freeze-dried and
stored in a refrigerator. Six hundred milligrams of the sample
was washed with CHCl 3/MeOH (2:1) and air-dried. One
hundred and fifty milligrams of the dried sample was homog-
enized with 8.0 mL of 0.01 M phosphate buffer (pH 6.0) and
centrifuged (3000 rpm, 10 min). The supernatant was used as
the sample.
Determination of the browning rate of L-Lys samples
containing saccharide and NaCl The color intensity of the
reaction products was measured with a UV/VIS spectropho-
tometer DU650 (Beckman Coulter, Fullerton, CA) at 470
nm.
Determination of furosine in L-Lys samples and meat
samples containing saccharide and NaCl The amount of
69
Sucrose (%) NaCl (%)
0.0 0.0
0.0 1.0
0.0 0.0
0.0 0.0
10.0 0.0
5.0 0.0
0.0 1.0
0.0 1.0
10.0 1.0
5.0 1.0
Amadori product was determined by the furosine content
because ε-N-deoxy-fructosyl-lysine, the major Amadori
product, is converted to furosine in an acid hydrolysis (Faist
et al., 2001). Quantification of furosine was carried out us-
ing the method of Nicoletti et al. (2000), with slight modi-
fication. Freeze-dried powder (L-Lys samples contained 90
mg of the L-Lys or meat samples contained 50 mg of the
protein) was subjected to acid hydrolysis by the addition of
8.0 mL of 8.0 N HCl in a test tube (16 × 100 mm). The pro-
tein in the sample was determined by the Kjeldahl method.
After purging with nitrogen for 2 min, the tube was capped
and heated at 110℃ for 23 h. The hydrolyzed sample was
filtered through a membrane filter with a pore size of 0.2
mm. Filtered sample (0.5 mL) was loaded onto a solid-phase
extraction column (Bond Elut-C18, 1.0 mL/100 mg, Varian
Associates, Harbor City, CA), washed with 1.0 mL of MeOH
and 3.0 mL of distilled water, and then extracted with 0.5 mL
of 8.0 N HCl. Analysis of furosine was done using a HPLC
−10Avp (Shimadzu, Kyoto, Japan) under the following con-
ditions: column, Inertsil Peptides C18 (4.6 × 250 mm, GL
Science, Tokyo, Japan); mobile phase, 5.0 mM sodium hep-
tanesulphonate with 20% acetonitrile and 0.2% formic acid;
flow rate, 1.0 mL/min; and detection at 280 nm. The furosine
standards (0.5, 1.0, 5.0, 10.0, 50.0 μg/mL), after filtering
through a membrane filter with a pore size of 0.2 mm, af-
forded the linear calibration curves (R2 = 0.9998).
Determination of fluorescent compounds in meat samples
Fluorescent compounds in meat samples were measured
using a Shimadzu RF-1500 spectrofluorophotometer. Most
fluorescent compounds of the Maillard reaction have typical
excitation wavelengths of 340 − 370 nm and emission wave-
lengths of 420 − 470 nm (maximum 370/440 nm for exc./em.)
(Matiacevich et al., 2005). Therefore, emission fluorescence
was scanned from 220 to 900 nm with excitation at 340 nm.
Then, the emission was fixed at 425 nm because the maxi-
70
mum peak was observed at this wavelength. The excitation
was scanned from 220 to 900 nm. The protein concentra-
tions of the samples were measured by the Kjeldahl method.
Samples were dissolved in 0.2 M phosphate buffer (pH 6.0),
adjusted to 5.0 mg/mL. The samples were diluted to 10 − 35
times and measured at 333/425 nm (exc./em.).
Statistical analysis Data are shown as the mean ± SD.
All calculations and statistical analyses were performed us-
ing STAT VIEW for Windows software (SAS Institute, Cary,
NC). Comparisons of the variables between the groups were
assessed by ANOVA. P values of less than 0.05 were consid-
ered significant. Tukey-Kramer was used as a post-hoc test.
Results and Discussion
Effect of seasoning on the browning of L-Lys The ef-
fects of saccharide and NaCl on the browning reaction of
L-Lys were investigated (Fig. 1). The sample containing su-
crose showed a very low browning rate. The Maillard reac-
tion is a chemical reaction between an amino acid and a re-
ducing sugar, and usually requires heating. Sucrose is a non-
reducing sugar and therefore does not react with L-Lys in the
Maillard-type browning reaction. On the other hand, absor-
bance at 470 nm of the sample containing D-Glc (a reducing
sugar) increased very rapidly during the reaction, and the ad-
dition of 1.0% NaCl significantly decreased the browning re-
action after 120 min. The result of the L-Lys sample contain-
ing D-Glc is exactly the same as the results we reported in a
previous study (Yamaguchi et al., 2009). The concentration
of NaCl was lower than in previous reports; however, the ad-
20
15
Absorbance at 470 nm
10
5 Lys-Glc-NaCl
0
0 30
Fig. 1. Degree of browning of L-Lys samples containing saccharide and NaCl.
The reaction conditions are described in “Materials and Methods”. Each value is the mean of 3 determinations.
*p < 0.05 by Tukey-Kramer post-hoc test.
K. Yamaguchi et al.
dition of NaCl also decreased the browning rate reaction in
this experiment. The reason for this is not clearly understood.
Amino acids are ampho ions in water. Dissociation constants
of amino acid residues in protein seemed to increase with the
addition of NaCl, and thus the rate of the Maillard reaction
decreased.
Effect of seasoning on furosine formation from L-Lys
The effects of saccharide and NaCl on the amount of furosine
formed from L-Lys were investigated (Fig. 2). Furosine was
not detected in the L-Lys only sample or the L-Lys sample
containing NaCl without saccharide. The sample containing
L-Lys and sucrose showed an extremely low amount of furo-
sine. On the other hand, the amounts of furosine in samples
containing L-Lys and D-Glc increased until 60 min, and
decreased thereafter. The reason for this is the progression
of the Maillard reaction, in which fructosyl-lysine is formed
until 60 min, leading to the formation of intermediates and
end products. Hartkopf (1993) investigated a model experi-
ment with a lysine-Glc mixture, heated at 100℃ for up to
30 h. The amount of furosine in the mixture increased until
24 h, and decreased thereafter. Addition of 1.0% NaCl to the
L-Lys sample containing D-Glc significantly decreased the
formation of furosine at 60 min. Therefore, it was suggested
that the addition of NaCl has an effect on the early stage of
the Maillard reaction in the case of amino acid with D-Glc.
Effect of seasoning on furosine formation in broiled meat
Pompei and Spagnolello (1997) reported that furosine con-
tent in tumbled meat containing NaCl and Glc was signifi-
cantly higher than that in raw meat. However, the common
Lys
Lys-NaCl
Lys-Glc
Lys-Suc
Lys-Suc-NaCl
60 90 120 150
Time （min）
Markers of the Maillard Reaction of Meat 71

Fig. 2. Amounts of furosine in L-Lys samples containing saccharide and NaCl.

The reaction conditions are described in “Materials and Methods”. Each value is the mean of 6 determinations.
*p < 0.05 by Tukey-Kramer post-hoc test.

Fig. 3. Amounts of furosine in pan-broiled (at 200°C) meat samples containing saccharide and NaCl.
The reaction conditions are described in “Materials and Methods”. The explanation of figure legends is shown in Table 2.
Each value is the mean of 6 determinations. *p < 0.05 by Tukey-Kramer post-hoc test. The concentration of NaCl was 1.0%.

sugar used as a seasoning, sucrose, is a non-reducing sugar.
Therefore, we tested sucrose as a saccharide. Results of furo-
sine of pan-broiled meat samples containing saccharide (D-
Glc or sucrose) and NaCl are shown in Fig. 3. The amounts
of furosine in the meat only sample and the meat sample con-
taining NaCl showed low values, but not zero, as the meat
itself contains a small amount of Glc. The sample containing
sucrose showed similar results. The amounts of furosine in
samples containing 5.0% and 2.5% D-Glc increased very
rapidly during the reaction. The addition of 1.0% NaCl in the
sample containing 5.0% D-Glc significantly decreased the
formation of furosine, but the decrease was not observed in
the sample containing 2.5% D-Glc. The common sugar used
as a seasoning is a non-reducing sugar, sucrose; therefore, it
was suggested that the contribution of sucrose and NaCl to
the formation of furosine was rather small in the case of pan-
72 K. Yamaguchi et al.

broiled meat samples.
Effects of cooking methods on furosine formation in
meat In actual cooking, there are various heat settings.
Hence, furosine content and internal temperature in the pan-
broiled, baked and fried meat samples were measured (Fig.
4). The final weights of meat samples without seasoning
were 30.2 g, 30.8 g and 16.7 g for the pan-broiled, baked and
fried meat samples, respectively. On the other hand, the final
weights of meat samples with seasoning were 30.7 g, 35.2
g and 20.2 g for pan-broiled, baked and fried meat, respec-

Fig. 4. Amounts of furosine and internal temperature of pan-broiled (A), baked (B) and fried (C) meat
samples containing white superior soft sugar and NaCl.
The reaction conditions are described in “Materials and Methods”. Each value is the mean of 6 determinations.
*p < 0.05 by Tukey-Kramer post-hoc test. Meat was added with 5.0% white superior soft sugar and 1.0% NaCl.
Markers of the Maillard Reaction of Meat
tively. The furosine content in the pan-broiled meat sample
(Fig. 4A) and fried meat sample (Fig. 4C) increased linearly
until 10 min, but the amount of furosine in the baked meat
sample (Fig. 4B) did not increase until 15 min and increased
rapidly after 15 min. It seems that the reason for this was
that the internal temperature of the meat samples increased
differently depending on the cooking method. In the case of
pan-broiled and fried meat samples, the internal temperature
increased rather rapidly compared to the baked meat sample.
Therefore, the furosine content in pan-boiled and fried meat
samples increased more than that in the baked meat sample
at the early stage of the Maillard reaction. It was also sug-
gested that the low water content of fried meat sample pro-
motes the furosine formation, judging from the final weight
of the meat sample. However, it was thought that the reason
why the amount of furosine in the pan-broiled and fried meat
samples did not reach more than 120 mg/100 g protein was
that the internal temperature of the meat samples did not in-
crease to more than 100℃. These results showed that the for-
mation of furosine varies depending on the cooking method.
The addition of seasonings, not glucose, to baked and fried
meat samples significantly increased the amount of furosine.
However, only a slight difference was observed between the
tested cooking methods. Therefore, the contribution of white
superior soft sugar and NaCl to the formation of furosine was
small for all three cooking methods. Each sample was burnt
by 30 min, but the internal color of the meat was still grey.
Formation of fluorescent compounds during meat cook-
Fig. 5. Comparison of fluorescence intensity of meat samples heated by three types of cooking methods.
The reaction conditions are described in “Materials and Methods”. Fluorescence was measured with 370/440 nm (exc./em.).
Each value is the mean of 3 determinations. *p < 0.05 by Tukey-Kramer post-hoc test.
73
ing Most fluorescent compounds produced in the Maillard
reaction have typical excitation wavelengths of 340 − 370 nm
and emission wavelengths of 420 − 470 nm (Matiacevich et
al., 2005), and the fluorescence in vivo is usually measured
at 370/440 nm (exc./em.) (Forbes et al., 2004) or 350/450
nm (exc./em.) (Ahmed and Thornalley, 2007). Therefore,
the effect of different cooking methods on the formation of
fluorescent compounds was investigated (Fig. 5 − 7). The
fluorescence intensity of the baked meat sample (Fig. 5 ○),
measured by wavelengths of 370/440 nm (exc./em.), did
not increase. On the other hand, the fluorescence intensities
of pan-broiled (Fig. 5 □) and fried meat samples (Fig. 5 △)
increased during heating. The fluorescence intensity of the
baked meat sample was significantly lower than that of the
samples heated by other cooking methods after 10 min of
heating.
The excitation wavelength was fixed at 340 nm, and
emission was scanned from 220 to 900 nm. The maximum
peak was observed at 425 nm (data not shown). Then, the
emission wavelength was fixed to 425 nm, and the excitation
was scanned from 220 to 900 nm (Fig. 6). The maximum
emission peak was observed at 333 nm in the pan-broiled
meat sample (Fig. 6B). The same results were obtained for
the baked and fried meat samples (data not shown). How-
ever, this peak was not observed in the raw meat sample (Fig.
6A). The wavelengths of excitation and emission for meat
samples were lower than previous reports (Forbes et al.,
2004; Ahmed and Thornalley, 2007). Therefore, the increase
74 K. Yamaguchi et al.

A B

Fig. 6. Fluorescence spectrums of raw meat sample (A) and pan-broiled meat sample (B) at 200℃ for 30 min.
The emission wavelength was fixed at 425 nm, and excitation wavelength was scanned from 220 to 900 nm.

35000

30000
Fluorescence intensity

25000

20000 Pan-broiled meat

Baked meat
15000
Fried meat
10000

5000

0
0 5 10 15 20 25 30
Time (min)

Fig. 7. Comparison of fluorescence intensity of meat samples heated by three types of cooking methods.
The reaction conditions are described in “Materials and Methods”. Fluorescence was measured with 333/425 nm (exc./em.).
Each value is the mean of 3 determinations. *p < 0.05 by Tukey-Kramer post-hoc test.

in fluorescence intensity at 333/425 nm (exc./em.) was mea-
sured during heating (Fig. 7). A similar trend was observed
at 370/440 nm, but the fluorescence intensity at 333/425 nm
was 300 times higher than that at 370/440 nm. To investi-
gate the fluorescent compounds in cooked meat, fluorescent
compounds with fluorescence at 333/425 nm were more
sensitive to detection than the usual Millard reaction fluores-
cent compounds, which have fluorescence at 370/440 nm or
350/450 nm. Ahmed and Thornalley (Ahmed and Thornalley,
2007) reported that the best method to detect the Maillard
reaction products is the LC/MS/MS method. However, this
method has not yet been established, and there are significant
problems and limitations associated with AGE immunoassay
procedures. Therefore, the authors recommended using “total
AGE fluorescence” to investigate AGE formation. Leclère
and Birlouez-Aragon (2001) analyzed lysine damage upon
heating using milk-resembling model systems. Heating at
60 − 85℃ first induced a parallel increase in furosine and ad-
vanced Maillard product (AMP) fluorescence. Later, furosine
reached a steady-state concentration, whereas AMP fluores-
cence continued to increase, showing a correlation with ly-
sine blockage. The authors concluded that AMP fluorescence
is a good indicator of lysine damage due to heating of milk in
the early and advanced stages of the Maillard reaction. In our
study, since the characteristics of fluorescent products from
the meat sample were observed to be similar to those of the
Markers of the Maillard Reaction of Meat
milk model systems, it was suggested that fluorescence mea-
surement was an effective indicator to assess the progress of
the Maillard reaction in foods.
In this study, each meat sample was heated for 30 − 60
min to a burnt state to investigate the production of furosine
and fluorescent compounds. However, a heating time of
10 − 15 min seems to be the optimal time for food quality and
also food safety by reducing the formation of harmful Mail-
lard reaction products.
References
Ahmed, N. and Thornally, P. J. (2007). Advanced glycation end-
products: what is their relevance to diabetic complications? Dia-
betes Obes. Metab., 9, 233-245.
Becalski, A., Lau, B.P.Y., Lewis, D. and Seaman, S.W. (2003).
Acrylamide in foods: occurrence, sources, and modeling. J. Ag-
ric. Food Chem., 51, 802-808.
Bosch, L., Alegri, A., Farre, R. and Clemente, G. (2008). Effect of
storage conditions on furosine formation in milk-cereal based
baby foods. Food Chem., 107, 1681-1686.
Brownlee, M. (2001). Biochemistry and molecular cell biology of
diabetic complications. Nature, 414, 813-820.
Cheng, K.W., Chen, F. and Wang, M. (2006). Heterocyclic amines:
chemistry and health. Mol. Nutr. Food Res., 50, 1150-1170.
Chiang, G.H. (1968). A simple and rapid high-performance liquid
chromatographic procedure for determination of furosine, a ly-
sine reducing sugar derivative. J. Agric. Food Chem., 31, 1373-
1374.
Delgado-Andrade, C., Rufiän-Henares, J.A. and Morales, F.J.
(2007). Lysine availability is diminished in commercial fibre en-
riched breakfast cereals. Food Chem., 100, 725-731.
Erbersdobler, H.F., Dehn-Mller, B., Nangpal, A. and Reuter, H.
(1987). Determination of furosine in heated milk as a measure of
heat intensity during processing. J. Dairy. Res., 54, 147-151.
Erbersdobler, H.F. and Hupe, A. (1991). Determination of lysine
damage and calculation of lysine bio-availability in several pro-
cessed foods. Z. Ernhrungswissenschaft, 30, 46-49.
Faist, V., Müller, C., Drusch, S. and Erbersdobler, H.F. (2001).
Selective fortification of lysinoalanine, fructoselysine and N
epsilon-carboxymethyllysine in case in model systems. Nahrung,
45, 218-221.
Finot, P.A., Bricout, J., Viani, R. and Mauron, J. (1968). Identifica-
tion of a new lysine derivative obtained upon acid hydrolysis of
heated milk. Experientia, 24, 1097-1099.
Forbes, J.M., Yee, L.T., Thallas, V., Lassila, M., Candido, R., Jan-
deleit-Dahm, K.A., Thomas, M.C., Burns, W.C., Deemer, E.K.,
Thorpe, S.R., Cooper, M.E. and Allen, T.J. (2004). Advanced
glycation end product interventions reduce diabetes-accelerated
atherosclerosis. Diabetes, 53, 1813-1823.
Garcia-Banos, J.L., Corzo, N., Sanz, M.L. and Olano, A. (2004).
75
Maltulose and furosine as indicators of quality of pasta products.
Food Chem., 88, 35-38.
Gerrard, J.A. (2002). Protein-protein crosslinking in food: Methods,
consequences, applications. Trends Food. Sci. Tech., 13, 391-399.
Hartkopf, J. (1993). N-ecarboxymethyl-lysin als Hitzeschädigungs-
indikator – Modell-untersucrosehungen zur Bildung in Lebens-
mitteln. Thesis, University of Kiel, Kiel.
Henle, T., Schwarzenbolz, U. and Klostermeyer, H. (1997). Detec-
tion and quantification of pentosidine in foods. Food Res. Tech.,
204, 95-98.
Heyns, K., Heukeshoven, J. and Brose, K.-H. (1968). Der Abbau
von Fructose-Aminosäuren zu N-(2-Furoylmethyl)-Aminosäuren.
Zwischenprodukte der Bräunungsreaktionen. Angew. Chem., 80,
627.
Hofmann, S.M., Dong, H.J., Li, Z., Cai, W., Altomonte, J., Thung,
S.N., Zeng, F., Fisher, E.A. and Vlassara, H. (2002). Improved
insulin sensitivity is associated with restricted intake of dietary
glycoxidation products in the db/db mouse. Diabetes., 51, 2082-
2089.
Knize, M.G. and Felton, J.S. (2005). Formation and human risk of
carcinogenic heterocyclic amines formed from natural precursors
in meat. Nutr. Rev., 63, 158-165.
Leclère, J. and Birlouez-Aragon, I. (2001). The fluorescence of
advanced Maillard products is a good indicator of lysine damage
during the Maillard reaction. J. Agric. Food Chem., 49, 1682-
1687.
Matiacevich, S.B., Santagapita, P.R. and Buera, M.P. (2005). Fluo-
rescence from the maillard reaction and its potential applications
in food science. Crit Rev Food Sci Nutr., 45, 483-495.
Moon, J.K. and Shibamoto, T. (2011). Formation of carcinogenic
4(5)-methylimidazole in maillard reaction systems. J. Agric.
Food Chem., 59, 615-618.
Mottram, D.S., Wedzicha, B.L. and Dodson, A.T. (2002). Acryl-
amide is formed in the Maillard reaction. Nature, 419, 448-449.
Murkovic, M., Wber, H.-J., Geiszler, S., Frohlich, K. and
Pfannhauser, W. (1999). Formation of the food associated carcin-
ogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)
in model systems. Food Chem., 65, 233-237.
Nicoletti, I., Cogliandro, E., Corradini, C., Corradini, D. and Piz-
zoferrato, L. (2000). Determination of furosine in hydrolyzate of
processed milk by HPLC using a narrow bore column and diode-
array detector. J. Liq. Chromatogr. Rel. Technol., 23, 717-726.
Pompei, C. and Spagnolello, A. (1997). Furosine as an index of heat
treatment intensity in meat products: Its application to cooked
ham. Meat Sci., 46, 139-146.
Resmini, P., Pellegrino, L. and Batelle, G. (1990). Accurate quanti-
fication of furosine in milk and dairy products by a direct HPLC
method. Ital. J. Food Sci., 3, 173-183.
Sanz, M.L., Del Castillo, D.M., Corzo, N. and Olano, A. (2003).
2-Furoylmethyl amino acids and hydroxymethylfurfural as indi-
76
cators of honey quality, J. Agric. Food Chem., 51, 4278-4283.
Sell, D.R. and Monnier, V.M. (1989). Structure elucidation of a
senescence crosslink from human extra-cellular matrix. J. Biol.
Chem., 264, 21597-21602.
Shibamoto, T. (1983). Heterocyclic compounds in browning and
browning/nitrite model systems: Occurrence, formation mecha-
nisms, flavor characteristics and mutagenic activity. In “Instru-
mental Analysis of Foods.” ed. by G. Charalambous and G. In-
glett, G. Academic Press, New York, pp. 229-278.
Stadler, R.H., Blank, I., Varga, N., Robert, F., Hau, J., Guy, P.A.,
Robert, M.C. and Riediker, S. (2002). Acrylamide from Maillard
reaction products. Nature, 419, 449-450.
Sugimura, T., Wakabayashi, K., Nakagama, H. and Nagao, M.
(2004). Heterocyclic amines Mutagens/carcinogens produced
during cooking of meat and fish. Cancer Sci., 95, 290-299.
Tirelli, A. and Pellegrino, L. (1995). Determination of furosine in
dairy products by capillary zone electrophoresis; a comparison
with the HPLC method. Ital. J. Food Sci., 7, 379-385.
Tokusoglu, O., Akalin, A.S. and Unal, K. (2006). Rapid high per-
formance liquid chromatographic detection of furosine (ε-N-2-
furoylmethyl-L-lysine) in yogurt and cheese marketed in Turkey.
K. Yamaguchi et al.
J. Food Qual., 29, 38-46.
Turesky, R.J. (2007). Formation and biochemistry of carcinogenic
heterocyclic aromatic amines in cooked meats. Toxicol. Lett.,
168, 219-227.
Uribarri, J., Cai, W., Sandu, O., Peppa, M., Goldberg, T. and Vl-
assara, H. (2005). Diet-derived advanced glycation end products
are major contributors to the body’s AGE pool and induce inflam-
mation in healthy subjects. Ann. N.Y. Acad. Sci., 1043, 461-466.
Villamiel, M., Arias, M., Corzo, N. and Olano, A. (2000). Survey of
the furosine content in cheeses marketed in Spain. J. Food Prot.,
63, 974-975.
Vlassara, H., Cai, W., Crandall, J., Goldberg, T., Oberstein, R.,
Dardaine, V., Peppa, M. and Rayfield, E.J. (2002). Inflammatory
mediators are induced by dietary glycotoxins, a major risk factor
for diabetic angiopathy. Proc. Natl. Acad. Sci. U.S.A., 99, 15596-
601.
Yamaguchi, K., Noumi, Y., Nakajima, K., Nagatsuka, C., Aizawa,
H., Nakawaki, R., Mizude, E., Otsuka, Y., Homma, T. and Chuy-
en, N. V. (2009). Effects of salt concentration on the reaction rate
of Glc with amino acids, peptides, and proteins. Biosci Biotech-
nol Biochem., 73, 2379-2383.