Food Chemistry 214 (2017) 300307

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Analytical Methods

Capillary electrophoresis method with UV-detection for analysis of free

amino acids concentrations in food
Mei Musa Ali Omar a,b,c, Abdalla Ahmed Elbashir b,, Oliver J. Schmitz a,
a

Applied Analytical Chemistry, Faculty of Chemistry, University of Duisburg-Essen, Essen, Germany

Department of Chemistry, Faculty of Science, University of Khartoum, Khartoum 11115, Sudan
c
Central Laboratory, Ministry of Sciences & Technology, P. O. Box Office 7099, Khartoum, Sudan
b

a r t i c l e

i n f o

Article history:
Received 11 March 2016
Received in revised form 14 June 2016
Accepted 9 July 2016
Available online 13 July 2016
Keywords:
Amino acids
Capillary electrophoresis
4-Chloro-7-nitro-2,1,3-benzoxadiazole
(NBD-Cl)
Food

a b s t r a c t
Simple and inexpensive capillary electrophoresis with UV-detection method (CE-UV) was optimized and
validated for determination of six amino acids namely (alanine, asparagine, glutamine, proline, serine and
valine) for Sudanese food. Amino acids in the samples were derivatized with 4-chloro-7-nitro-2,1,3-ben
zoxadiazole (NBD-Cl) prior to CE-UV analysis. Labeling reaction conditions (100 mM borate buffer at pH
8.5, labeling reaction time 60 min, temperature 70 C and NBD-Cl concentration 40 mM) were systematically investigated. The optimal conditions for the separation were 100 mM borate buffer at pH 9.7 and
detected at 475 nm. The method was validated in terms of linearity, limit of detection (LOD), limit of
quantification (LOQ), precision (repeatability) (RSD%) and accuracy (recovery). Good linearity was
achieved for all amino acids (r2 > 0.9981) in the concentration range of 2.540 mg/L. The LODs in the
range of 0.320.56 mg/L were obtained. Recoveries of amino acids ranging from 85% to 108%, (n = 3) were
obtained. The validated method was successfully applied for the determination of amino acids for
Sudanese food samples.
2016 Elsevier Ltd. All rights reserved.

1. Introduction
Amino acids are important for life because they are the basic
components of proteins and serve as a source of energy (Cui
et al., 2014; Veledo, de Frutos, & Diez-Masa, 2005). The nutritional
value of proteins depends mainly on their amino acids composition
(Gonzlez-Castro, Lpez-Hernndez, Simal-Lozano, & OrunaConcha, 1997; Veledo et al., 2005). Rapid and efficient determination of amino acids concentration in complex matrices is of broad
interest in food chemistry and industry. Determination of free
amino acids levels play a major role for the quality and safety of
many foods (Veledo et al., 2005).
Determination of amino acids in complex samples is a challenge
because they are not sufficiently volatile, most of them are highly
polar and do not have a chromophore (Elbashir, Aboul-Enein, &
Suliman, 2011; Lorenzo, Navarrete, Balderas, & Garcia, 2013).
Several techniques have been described for the determination
of free amino acids, including high performance liquid chromatography (HPLC), gas chromatography (GC) and capillary electrophoresis (CE) (Cui et al., 2014; Mustafa, man, Andersson, &
Corresponding authors.
E-mail addresses: aaelbashir@uofk.edu, hajaae@yahoo.com (A.A. Elbashir),
oliver.schmitz@uni-due.de (O.J. Schmitz).
http://dx.doi.org/10.1016/j.foodchem.2016.07.060
0308-8146/ 2016 Elsevier Ltd. All rights reserved.

Kamal-Eldin, 2007; Song, Funatsu, & Tsunoda, 2013). The traditional analytical technique used to measure free amino acids is
cation-exchange chromatography with post-column derivatization
with ninhydrin (Lorenzo et al., 2013). However, these methods
have drawbacks, such as they require expensive equipment, long
analysis time and extensive sample preparation and cleanup
(Mustafa et al., 2007; Soga & Heiger, 2000; Warren, 2008). Reverse
phase HPLC (RP-HPLC) methods for amino acids analysis suffered
from poorly retained of polar amino acids on the RP columns and
difficult to separate from the solvent peak (Warren, 2008). CE
has been considered as a powerful separation technique for amino
acids and peptides in complex samples (Poinsot, Bayle, & Couderc,
2003; Akamatsu & Mitsuhashi, 2013; Hirayama, Igarashi, Tomita, &
Soga, 2014; Simionato, Moraes, Carrilho, Tavares, & Kenndler,
2008).
UVvis detector is the most used detector in commercially
available CE system because of its general applicability (Zacharis
et al., 2006). Since most of amino acids have no chromophore, a
derivatization step is often essential in order to enhance their
detectability using optical detection (Neda et al., 2012). As it has
been reviewed in literature the commonly used pre-column
derivatizing reagents for amino acids include 6-aminoquinoly-Nhydroxysuccimidyl carbamate (AQC), o-phthalaldehyde (OPA),
naphthalene
dicarboxaldehyde
(NDA),
9-fluorenylmethyl

M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

301

Fig. 1. Electropherograms obtained at different derivatization pH. Amino acids concentration 10 mg/L each. Derivatization conditions: NBD-Cl concentration 30 mM, reaction
time 40 min, 100 mM borate buffer, reaction temperature 60 C. Peaks: Pro, proline; Val, valine; Gln, glutamine; Ala, alanine; Asn, asparagine; Ser, serine.

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M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

Fig. 5. Effects of the pH of running buffer on the separation of six amino acid mixtures after derivatization with NBD-Cl. CE conditions: 100 mM borate buffer with the pH of
the running buffer changed from 8.5 to 10; separation voltage 25 kV; injection sample 10 s under the pressure of 0.5 psi; capillary temperature set at 25 C.

M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

chloroformate (FMOC-Cl), phenylisothiocyante (PITC) and dansyl

chloride (Kang, Xiao, Huang, & Gu, 2006; Salazar, Armenta, &
Shulaev, 2012;). However these derivatization agents have some
drawbacks such as some of their derivatives are sensitive to moisture and instable (OPA) (Bani Rashaid, Jackson, & Harrington,
2014). AQC amino acids derivatives required long analysis time
(more than 45 min) (Mayer & Fiechter, 2013) and high solvent consumption in addition commonly used with fluorescence detector.
The main limitations of use of OPA and NDA as derivatization
reagents are: they do not react with secondary amino acids, some
of OPA amino acids derivatives are unstable and NDA react only
with primary amino acids in the presence of toxic compounds such
as sodium cyanide (Lorenzo et al., 2013; Zhang & Sun, 2004). The
disadvantage of using of FMOC-Cl and PITC as a pre-column derivatization reagents is the hydrolysis and the production of byproducts which interfered in the analysis unless the excess of
reagents are removed prior to the analysis (Gonzlez-Castro
et al., 1997). Dansyl chloride lacking the selectivity and it reacts
with both OH and NH2 groups (Gonzlez-Castro et al., 1997).
4-Chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) is a labeling
agent that have been used for the determination of primary and
secondary amino acids by pre column derivatization (Elbashir,
Krieger, & Schmitz, 2014). NBD-Cl has merits such as low cost, it
can be used for fluorescent labeling of amino acids and UV detection, and this reagent produces low number of byproducts
(Lorenzo et al., 2013).
Most of CE methods based on using NBD-Cl as a pre-column
derivatization agent for amino acids employed laser induced fluorescence (LIF) detector (Cui et al., 2014; Elbashir et al., 2011). These
methods resulting in low detection limits, however, the concentration of amino acids are high in foods with the highest proteins content. Moreover LIF detector is more expensive than UV and PDA
detector and is not available in many laboratories. Due to availability CE system with PDA detector, this current work devoted to optimize and validate CE with PDA detector for analysis six amino
acids namely (alanine, asparagine, glutamine, proline, serine and
valine) in food samples after derivatization with NBD-Cl. It has
been reported that there is a relation between acrylamide which
was classified as a probable human carcinogen (Group 2A) content
and the quantities of amino acids namely, asparagine in food (Bent,
Maragh, & Dasgupta, 2012; Elmore, Briddon, Dodson, & Halford,
2015). HPLC method for analysis of amino acids in wheat and
chickpea flour was reported by Izembayeva, Bayisbayeva,
Muldabekova, Iztayev, & Dautkanova, 2014. The amino acid in
range of 4.181.43 mg/kg were measured. Jia et al. (2001) developed an ion chromatography method for analysis of some amino
acids and asparagine in soybean. The concentration in range of
asparagine was found in range of 1.618.4 lmol g 1. The amino
acids were analyzed in sorghum by ion-exchange chromatography
(Adeyeye, 2008). Therefore in this research paper a CE- with PDA
method was developed and validated and applied for quantification of amino acids content in Sudanese food samples in order to
correlate with the previously measured concentration of acrylamide in final products (Omar, Wan Ibrahim, & Elbashir, 2014
and Omar, Elbashir, & Schmitz, 2015).

2. Material and method

2.1. Instrumentation
All electrophoretic separations of amino acids were performed
on a Beckman P/ACE MDQ CE system with PDA detector. The
PDA was set at 475 nm. Separation was carried out on fusedsilica capillaries from Polymicro Technologies (Phoenix, AZ, USA)
of 40 cm total length (30 cm from inlet to the detector window)

303

with internal diameter (id) of 50 lm. The capillary was thermostated at 25 C. The new capillaries were first conditioned
with1.0 mol/L NaOH for 15 min, 0.1 mol/L NaOH for 5.0 min, water
for 5.0 min and the running buffer for 5.0 min. Before each injection, the capillary was preconditioned with, 0.1 mol/L NaOH, water
and the running buffer for 3.0 min each. Samples were injected by
pressure at 0.5 psi for 10 s, and separations were performed under
25 kV with a positive high voltage. The data were collected and
processed by Beckman P/A CE 32 Karat software Version 4.0.
2.2. Chemicals
Amino acids analytical standards (L-alanine (98%), L-asparagine
(98%), L-glutamine (99%), L-proline (99%), L-serine (99%) and Lvaline (98%) were purchased from Sigma (St. Louis, MO, USA).
NBD-Cl 98%, was obtained from sigma-Aldrich (Steinheim, Germany). Ethanol and acetonitrile of HPLC grade were got from
VWR PROLABO chemicals. Water was purified with arium pro
ultrapure water systems from Sartorius Stedim Biotech (Gttingen,
Germany). Boric acid and sodium hydroxide were obtained from
VWR International (Leuven, Belgium).
2.3. Preparation of the electrolytes and standard solutions
The running buffers (100 mM) were prepared by dissolved
0.6183 g boric acid in 80 mL of de-ionized water and the pH
adjusted into desired value with 1.0 M sodium hydroxide then
the volume was completed to 100 mL with de-ionized water. The
stock solutions of each amino acid at a concentration of
1000 mg/L were prepared in deionized water. Then the mixed
amino acids stock solution was prepared at concentration of
100 mg/L in 100 mM borate buffer (pH 8.5). All the working solutions in range 2.540 mg/L were obtained by diluting the mixed
solution with 100 mM borate buffer (pH 8.5). 100 mM of NBD-Cl
stock solution was prepared in acetonitrile and was diluted to
appropriate concentrations.
2.4. Samples
Five Sudanese samples namely potato, eggplant, chickpeas, soft
wheat flour, and sorghum durra flour were used as test samples
and were purchased from Sudanese local markets and produced
by Sayga Flour Mills Company, Khartoum North, and Sudan. All
food samples except wheat flour and sorghum durra flour were
pulverized and homogenized using household food processor
(Modern blender, China).
2.4.1. Extraction of free amino acids from food samples
Free amino acids in food samples were extracted as described
by Ohara-Takada et al. (2005) with some modifications. A homogenised sub-sample (3.0 g) of the selected food was weighed into a
50 mL centrifuge tube and 10 mL of 80% (v/v) ethanol was added.
The centrifuge tube was shaken vigorously for 1.0 min using a
vortex-2 Genie mixer (Bohemia, USA). Then the amino acids in
homogenate were extracted three times in mechanical shaker
(Bender & Hobein AG, Switzerland) at room temperature for one
hour. The extract was centrifuged in Allegra 25R centrifuge from
Beckman Coulter (California, USA) at 6028g (4000 rpm) for
5.0 min. Then the supernatant was passed through two layers of
filter paper. An aliquot (5.0 mL) of filtrate was transferred into a
glass vial and evaporated to dryness under a gentle stream of nitrogen gas. The residue was reconstituted with 100 lL of 100 mM
borate buffer pH 8.5, vortex for 1.0 min and then samples were
subjected to derivatization.

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2.5. Optimized pre-column derivatization with NBD-Cl of standard

amino acids and free amino acids in food samples

3. Results and discussion

In total, 100 lL of 40 mM NBD-Cl solution in acetonitrile was

added to 100 lL of sample extracts or amino acids standards in
100 mM borate buffer pH 8.2. The mixture was vortex and incubated in thermomixer comfort (Eppendorf AG, Germany) at 70 C
for 60 min in the dark. The reaction was terminated by cooling in
ice water. Then the reaction mixture was filtered through a
0.2 llm Phenex PTFE syringe filter from Phenomenex (Torrance,
CA, USA) into a CE auto sampler vial for analysis.

2.6. Validation study

This method was validated in terms of linear range, limit of
detection (LOD), limit of quantification (LOQ), precision (repeatability) (RSD%) and accuracy (recovery). For linearity study, mixed
standards solutions of amino acids at five different concentration
levels ranging from 2.5 to 40 mg/L were derivatized with NBD-Cl
and then used to construct the calibration curves using optimum
conditions. The LOD and LOQ were obtained by multiplying the
standard deviation of the noise by 3.0 and 10, respectively. The
precision of this method was evaluated by intra-day repeatability
as RSD% for migration time and peak area. It was studied by performing five successive injections of 7.5 mg/L amino acid standards
mixture. For recovery study, the selected food samples with known
amino acid concentration was spiked with mixed amino acid
standards at concentration level of 10 mg/L. an aliquot of 1.0 mL
(100 mg/L) amino acids mixed standard was added into each food
sample and 9 mL of 80% (v/v) ethanol was added. The procedure
was continued as in Sections 2.4.1 and 2.5. Three replicates were
performed for each recovery level.

3.1. Optimization of derivatization reaction conditions

The optimum conditions for the method developed were established by varying the parameters one at a time while keeping the
others fixed and observing the effect produced on the peak area
of the colored product. In order to achieve higher sensitivity, various parameters affected on derivation reaction between NBD-Cl
and amino acids such as pH, reaction temperature, reaction time
and NBD-Cl concentration were studied.
3.1.1. Effect of pH on derivatization reaction
pH has a great effect on the reaction of amino acids with NBD-Cl
(Lorenzo et al., 2013; Zhang, Le Potier, Smadja, Zhang, & Taverna,
2006). The nucleophilic reaction between amino acids and
NBD-Cl took place under basic conditions. Thus various pH values
ranging from 7.5 to 9.5 of 100 mM borate buffer have been
investigated. The results show that the peak area of amino acids
derivatives increases with increasing the pH of borate buffer. However at pH value higher than 8.5, a broad peak with higher area of
byproduct NBD-OH appeared in the electropherogram (Fig. 1). This
result in an agreement with that reported by Zhang et al., 2006.
Thus pH of 8.5 was selected for derivatization reaction.
3.1.2. Effect of reaction temperature on derivatization
The effect of temperature on derivatization reaction was examined by varying temperature from 50 to 90 C. The results are
shown in Fig. 2 (supplementary material) indicated that the highest peak area was observed at 70 C for all amino acids derivatives
except for proline at 50 C. This suggested that the proline derivative with NBD-Cl is unstable at high temperature as proposed by
(Yang et al., 2007). The temperature of 70 C was selected as optimum derivatization temperature.

2.7. Data analysis

Statistical analyses were performed using Microsoft Excel professional plus 2010 (Microsoft corporation, Redmond, WA).

3.1.3. Effect of reaction time on derivatization

The effect of time in derivatization process was studied in range
from 10 to 80 min. It can be seen from Fig. 3 (supplementary

Table 1
Equations for standard addition calibration curves, regression coefficient, limit of detection (LOD) and limit of quantification (LOQ) for amino acids.

Amino acids

Equationa

Regression coefficient (r2)

LOD (mg/L)

LOQ (mg/L)

Proline
Valine
Glutamine
Alanine
Asparagine
Serine

Y = 37415x 41953
Y = 35199x 33389
Y = 8711.7x 2477
Y = 21712x 6997
Y = 11925x + 13457
Y = 12182x + 5940

0.9985
0.9990
0.9982
0.9986
0.9981
0.9994

0.32
0.36
0.49
0.41
0.53
0.56

1.06
1.89
1.62
1.35
1.75
1.85

RSD% (n = 5)
Migration time

Peak area

0.36
0.52
0.42
0.72
0.45
0.60

4.45
3.35
3.80
5.68
5.15
4.06

n = 2.

Table 2
Percentage recovery (n = 3) for determination of amino acids in Sudanese food samples.
Amino acids

Proline
Valine
Glutamine
Alanine
Asparagine
Serine

Added amount

Potato

Eggplant

Chickpeas

Wheat flour

Sorghum durra

(mg/L)

Found
(mg/L)

Recovery
(%) SD

Found
(mg/L)

Recovery
(%) SD

Found
(mg/L)

Recovery
(%) SD

Found
(mg/L)

Recovery
(%) SD

Found
(mg/L)

Recovery
(%) SD

10
10
10
10
10
10

9.8
10.8
9.2
9.0
9.4
9.5

98 3.0
108 6.3
92 4.5
90 2.1
94 3.6
95 1.8

9.5
9.2
8.8
8.5
9.0
8.9

95 5.4
92 3.3
88 3.8
85 4.2
90 2.9
89 2.2

9.3
9.5
9.8
9.2
9.4
9.5

93 0.9
95 1.7
98 2.4
92 3.6
94 1.5
95 3.1

9.6
9.9
10.2
9.3
10.1
9.7

96 4.8
99 2.9
102 4.6
93 3.8
101 3.2
97 5.7

9.3
9.0
9.1
8.7
9.4
9.0

93 2.3
90 1.9
91 2.6
87 2.3
94 3.6
90 1.95

M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

material), the peak area of amino acid derivatives increased with

increasing reaction time up to 60 min and the decreased for all
amino acids except for proline, with an optimum derivation time
of 40 min. The reaction time of 60 min was chosen as optimal time.

305

3.1.4. Effect of NBD-Cl concentration on derivatization

The influence of NBD-Cl concentration was investigated over
the range 1050 mM. As shown in Fig. 4 (supplementary material),
the peak area of amino acid derivatives increase with increasing

Fig. 6. Electropherograms of Chickpeas sample (A) unspiked (B) spiked with the six amino acids mixture at concentration level of 10 mg/L. CE conditions: 100 mM borate
buffer at pH of 9.7; separation voltage 25 kV; injection sample 10 s under the pressure of 0.5 psi; capillary temperature set at 25 C.

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M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

NBD-Cl concentration up to 40 mM, and then decrease slightly.

Therefore 40 mM was selected as optimum.
According to optimization studies, the optimized conditions
used for further studies were chosen as borate buffer pH 8.5, reaction temperature at 70 C, reaction time for 60 min and NBD-Cl
concentration of 40 mM.

Table 4
Concentrations of asparagine in Sudanese foods raw materials and of acrylamide in
Sudanese foods final products.

3.2. Optimization of separation conditions

In order to examine the separation conditions by CE with PDA
detector, a mixed standard of six amino acids at concentration
level of (7.5 mg/L) was derivatized with NBD-Cl and then used.
3.2.1. Effect of the pH of running buffer on separation
The pH of the running buffer affects the mobility of the analytes
in CE system by ionization of analytes and capillary wall. The effect
of pH of running buffer on separation of amino acid derivatives was
studied in the pH range of 8.510 as shown in Fig. 5. It can
observed that resolution of amino acid derivatives improved with
increasing buffer pH. The best separation of amino acids derivatives was obtained at buffer pH of 9.7, so it selected as optimum
pH for running buffer.
3.3. Validation of the methods
This method was validated in terms of linear range, limit of
detection (LOD), limit of quantification (LOQ), precision (repeatability) (RSD%) and accuracy (recovery).
3.3.1. Linearity, LOD and LOQ
The calibration curves were constructed by plotting amino acid
derivatives peak areas against concentrations of amino acids. As
shown in Table 1. Good linearities were obtained for all amino
acids with coefficient of determination (r2) exceed 0.9981 over
the concentration range studied. All data are summarized in
Table 1. The obtained LODs in this method are higher than LODs
obtained by using NBD-Cl as derivatization agent and LIF detection
(Shi, Liang, Song, Yang, & Gao, 2012). However these LODs are
suitable for determination of free amino acids in food samples,
since their concentration is high in food.
3.3.2. Precision and accuracy
It was studied by performing five successive injections of
7.5 mg/L amino acid standards mixture. Inter-day repeatability
results are summarized in Table 1. The RSD values of migration
time and peak area are less than 0.72 and 5.68% respectively, which
illustrate good precision.
The recovery test was carried out by spiking five Sudanese food
samples (potato, eggplant, chickpeas, soft wheat flour, and sorghum durra flour) with amino acids mix standard at concentration
level of 10 mg/L for each amino acid. The percentage recoveries
ranged from 85% to 108%, for all amino acids (Table 2).
3.3.3. Application of the method
The established method was assessed by analyzing free amino
acids in five Sudanese food samples (potato, eggplant, chickpeas,
soft wheat flour, and sorghum durra flour). The results are summarized in Table 3 (supplementary material). The values of the free
amino acids in potato and wheat flour are in agreement with the
ranges reported in the literature (Amrein, Andres, Escher, &
Amad, 2007; Borda & Alexe, 2011; Claeys, De Vleeschouwer, &
Hendrickx, 2005; Elahi & Khan, 1973; Lister & Munro, 2000;
Murniece et al., 2008; Prieto, Collar, & de Barber, 1990). For eggplant and chickpeas the values obtained are higher than reported
one (Flick, Ory, & St. Angelo, 1977; Izembayeva et al., 2014) where
as for sorghum durra flour amount less than the reported in the

Sample ID

Raw
material

Asparagine
(mg/100 g) SDa

Acrylamide
(lg/kg) SDa

Fried potato
Fried eggplant
Minnan
b.rae
Gorrasa
Taamia

Potato
Eggplant
Wheat flour
Sorghum durra
Wheat flour
Chickpeas

525 6.60
512 7.47
27 3.83
176 3.68
27 3.83
94 8.15

227 9.93
332.5 6.82
16.92 0.62
53.25 1.76
21.45 0.32
69.75 3.61

n = 3.

literature were obtained (Adeyeye, 2008; Claeys et al., 2005; Etuk

et al., 2012). The variation in the amount of the free amino acids
obtained may be due to the species, cultivar and growing conditions (Abdel-Aal & Hucl, 2002). The amino acid derivatives were
identified by comparing the migration times of amino acid standards and by standard addition methods. Fig. 6(A) and (B) shows
the typical electropherograms of amino acids from chickpeas sample before and after spiking at concentration level of 10 mg/L. It
was observed that the migration times of amino acid derivatives
are a little bit different in a real sample compare to standard solutions. This may be due to the complicated matrices of the real
samples.

3.3.4. Relationships between acrylamide formation and free amino

acids
Free asparagine was regarded as the most important precursor
for acrylamide formation (Bent et al., 2012; Elmore et al., 2015).
The amount of acrylamide that is formed is directly related to
the quantities of free amino acid asparagine that are present within
the foods (Bent et al., 2012). From Table 4, the highest acrylamide
concentrations were found in fried eggplant and fried potato
samples (332.5 and 227 lg/kg, respectively) (Omar et al., 2015),
which their raw materials contain the highest asparagine content
(512 and 525 mg/100 g, respectively). In the other analyzed
Sudanese food raw materials the concentration of free amino acid
asparagine is varied between (27 and 176 mg/100 g) which it did
not correlate well with acrylamide concentration in final products.
This mean the relationships between the concentration of acrylamide in the final products and the concentrations of asparagine
in starting materials are not clear and need more studies in
model systems.

4. Conclusion
An inexpensive CE-UV method with NBD-Cl as pre-column
derivatization agent was proposed for separation and determination of amino acids in food samples. Various derivatization
conditions were optimized and then the method was validated.
The methodology provides good linearity, precision and recoveries.
The method was applied to the analysis of six amino acids in Sudanese food samples raw materials. The obtained results for asparagine concentration in food raw material were correlated with
concentration of acrylamide in food final product measured in
our previous study Omar et al., 2015.

Conflict of interest
Author Mei Musa Ali Omar has received research grants from
DAAD.
All Authors declare that they have no conflict of interest.

M.M.A. Omar et al. / Food Chemistry 214 (2017) 300307

Ethical approval
This article does not contain any studies with animals
performed by any of the authors.
Informed consent
Not applicable.
Acknowledgment
The financial support from Deutscher Akademischer Austauschdienst (DAAD) is gratefully acknowledged.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2016.
07.060.
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