Food Chemistry 127 (2011) 1385–1390

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

Analytical Methods

Simultaneous analysis of acidulants and preservatives in food samples

by using capillary zone electrophoresis with indirect UV detection
Kenji Yoshikawa ⇑, Shintaro Saito, Akio Sakuragawa
Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University, 1-8-14, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8308, Japan

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

Article history: Capillary zone electrophoresis with indirect UV detection was developed for the simultaneous analysis of
Received 16 April 2010 acidulants and preservatives in food samples. When a solution of tris (hydroxymethyl) aminomethane,
Received in revised form 24 December 2010 trimellitic acid and poly (vinyl alcohol) was used as the background electrolyte, the nine acidulants
Accepted 26 January 2011
and four preservatives listed in the Japanese Food Sanitation Law were detected within 8 min. The cali-
Available online 1 February 2011
bration curves plotted from the peak height of each analyte were linear with a correlation coefficient of
0.99. The relative standard deviations (n = 10) of the peak height ranged from 1.2% to 4.7%. The detection
Keywords:
limits for these species ranged from 0.6 to 5.3 mg/L at a signal-to-noise ratio of three. The method devel-
Capillary zone electrophoresis
Acidulants
oped method was applied to the simultaneous analysis of acidulants and preservatives in a wide variety
Preservatives of food samples.
Indirect UV detection Ó 2011 Elsevier Ltd. All rights reserved.
Food samples

1. Introduction
In a wide variety of foods and beverages, organic acids in the
form of food additives, such as acidulants and preservatives, are
used (Chen & Ni, 2009; Han et al., 2008; Huang, Chuang, Chiu, &
Yen, 2005). For example, commonly in food products, citrate is
used as a chemical acidulant to add acidity and benzoate is used
as a chemical preservative to prevent alteration and degradation
by microorganisms during storage. Although excessive consump-
tion is undesirable, there are no restrictions on the use of acidu-
lants in Japan. However, excessive consumption of preservatives
may be harmful to the human body. The Japanese Food Sanitation
Law defines certain usage standards for preservatives (Ministry of
Health Labor and Welfare, 2007). Therefore, from the viewpoint of
food safety, appropriate qualitative and quantitative analysis of
these species in food is very important.
Acidulants and preservatives are typically analysed using de-
vices such as gas chromatography (GC) in combination with mass
detection techniques (Dong, Mei, & Chen, 2006; Wang, Zhang,
Wang & Wang; 2006), high-performance liquid chromatography
(HPLC) (Saad, Bari, Saleh, Ahmad, & Talib, 2005; Suárez-Luque,
Mato, Huidobro, Simal-Lozano, & Sancho, 2002; Shui & Leong,
2002), ion chromatography (IC) (Chen & Wang, 2001; Yoshikawa,
Okamura, Inokuchi, & Sakuragawa, 2007) and flow injection analy-
sis (FIA) (García-Jiménez, Valencia, & Capitán-Vallvey, 2007; Kim,
2006). As GC requires derivatization and complex sample pretreat-
⇑ Corresponding author. Tel.: +81 3 3259 0801; fax: +81 3 3293 7572.
E-mail address: yoshikawa@chem.cst.nihon-u.ac.jp (K. Yoshikawa).
ment, HPLC and IC are more commonly used. However, in case of
the detection of several analytes, multiple types of columns or gra-
dient methods must be used, and multicomponent analysis is
impossible with FIA. In addition, these species exhibit widely dif-
fering retentions that lead to long analysis times. Therefore, a
detection method addressing all these issues is desired.
Capillary electrophoresis (CE) is a powerful and flexible separa-
tion method because of its high resolution efficiency, low sample
volume and rapid analysis (Frazier, Inns, Dossi, Ames, & Nursten,
2000; Galli & Barbas, 2004; Mato, Suárez-Luque, & Huidobro,
2007; Soga & Ross, 1999; Tang & Wu, 2007). In CE, even if the sam-
ple contains a matrix, it can be injected with minimal sample prep-
aration without decreasing the separation performance. After the
sample analysis, the capillary is rapidly flushed with fresh back-
ground electrolyte (BGE) and is ready for the next injection. There-
fore, CE is a suitable analytical method for the analysis of food
sample containing various matrices.
Because the UV absorbance of common organic acids is poor,
most published studies on organic acid analysis by capillary zone
electrophoresis (CZE) have used indirect UV detection with various
BGEs. (Bord, Crétier, Rocca, Bailly, & Souchez, 2005; Cousins, Had-
dad, & Buchberger, 1994; Sam, Lalljie, Vindevogel, & Sandra, 1993).
Initially, we attempted to analyse the acidulants and preservatives
using CZE with direct UV detection. However, the migration time of
all the preservatives was found to be similar or exactly the same.
Despite changing the operating conditions, simultaneous analysis
of these species remained difficult. In addition, aromatic com-
pounds showed a tendency wherein the width of each peaks
extended.

0308-8146/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.01.126
1386 K. Yoshikawa et al. / Food Chemistry 127 (2011) 1385–1390
This study demonstrates the simultaneous analysis of acidu-
lants (citrate, tartrate, malate, succinate, adipate, acetate, lactate,
ascorbate and gluconate) and preservatives (dehydroacetate, sor-
bate, benzoate and propionate) by using CZE with indirect UV
detection. First, the operating conditions of these species (probe,
indirect UV detection wavelength, BGE conditions, capillary tem-
perature and applied voltage) were examined according to a pub-
lished method (Bord, Crétier, Rocca, Bailly, & Souchez, 2005).
Then, the effect of the presence of coexisting species (inorganic an-
ions and carbohydrates) in the food samples on the detection abil-
ity of the method and the analytical precision of these species were
examined. Finally, the developed method was applied to analyse
the acidulants and preservatives in several actual food samples.
2. Experimental
2.1. Apparatus
All CE experiments were performed on an HP3D CE system (Agi-
lent Technologies, Waldbronn, Germany), equipped with a diode
array detector, an automated sampler and a power supply capable
of delivering up to 30 kV. HP3D Chemstation software was used for
CE control, data acquisition and data handling.
2.2. Reagents
Acidulant (citrate, tartrate, malate, succinate, adipate, acetate,
lactate, ascorbate and gluconate) and preservative standard solu-
tions (dehydroacetate, sorbate, benzoate and propionate) were ob-
tained from Wako Pure Chemical (Osaka, Japan). Inorganic anion
standard solutions (bromide, chloride, nitrite, nitrate, sulphate,
sulphite and fluoride) were purchased from Wako Pure Chemical.
These standard solutions were prepared from their sodium salts,
potassium salts, or free acids. Carbohydrate standard solutions (su-
crose, fructose and glucose) were purchased from Wako Pure
Chemical. Each standard solution was prepared by diluting
2000 mg/L certified standards of acidulants, preservatives, inor-
ganic anions and carbohydrates. Tris(hydroxymethyl)aminometh-
ane (Tris; Wako Pure Chemical) and poly(vinyl alcohol) (PVA;
Aldrich, Steinheim, Germany) were used as BGEs. Phthalic acid
(PA; Wako Pure Chemical), trimellitic acid (TMA; Aldrich) and
pyromellitic acid (PMA; Tokyo Kasei, Tokyo, Japan) were used as
prove solutions. Hexadimethrine bromide (HDB; Aldrich) was ap-
plied as a coating solution. Hydrochloric acid (HCl) and sodium
hydroxide (NaOH) (Wako Pure Chemical) were used as pH adjust-
ers. All chemicals used in this study were of analytical grade purity.
Water was purified using a Milli-Q purification system (Millipore,
Bedford, MA, USA) with a specific resistance of 18.2 MX.
2.3. Operating conditions
All separations were performed on a bubble cell (previously ex-
tended light path) fused-silica capillary (Agilent Technologies,
Waldbronn, Germany) of 50 lm i.d.  64.5 cm (56 cm effective
length). Before each injection, the capillary was preconditioned
for three min by flushing with BGE. In addition, the capillary was
flushed for 10 min after use. The sample solution was injected at
a pressure of 50 mbar for 5.0 s. The BGE used contained 200 mM
Tris, 10 mM TMA and 0.10 (w/w)% PVA. Its pH was adjusted to
9.0 using 1 M HCl or 1 M NaOH. Electrophoretic separations were
performed at 25 °C and 30 kV (negative polarity). Indirect UV
detection was performed using a diode array detector. The signal
wavelength was set at 350 nm with a reference wavelength at
230 nm.
2.4. Preparation of BGE
BGE was prepared as follows: 0.10 g PVA was added to less than
100 mL water into a beaker and heated to 90 °C on a hot plate stir-
rer for 30 min. After the PVA had dissolved, the solution was cooled
to 25 °C. Next, 2.4228 g Tris and 0.2102 g TMA were added. The pH
was adjusted to 9.0 with 1 M HCl or 1 M NaOH. The resulting solu-
tion was prepared using a 100 mL measuring flask and filtered
using a 0.45 lm pore size filter before use.
2.5. Semi-permanent coating method of capillary
To reverse the electroosmotic flow (EOF), a bubble cell fused-
silica capillary semi-permanently coated with HDB was used. At
first, the fused-silica capillary was flushed with 0.1 M NaOH for
10 min to dissociate the silanol group in the inner surface of the
capillary. To coat the capillary, it was flushed with a 10 (w/w)%
HDB solution for 30 min. Next, it was washed with distilled water
for 25 min and with BGE for 10 min. Finally, it was flushed with
BGE and conditioned under 30 kV for 10 min before analysis. In
view of the baseline stability and the migration time of each ana-
lyte, this semi-permanent coating capillary was stable and suitable
for 100 injections.
2.6. Food samples
The food samples were red and white wine (Mercian Co. Ltd.), a
health drink (Takeda Pharmaceutical Co. Ltd.), a yogurt (Meiji dair-
ies Co. Ltd.) and the pickled ginger (Iwashita Shokuhin Co. Ltd.).
These samples were purchased at a local supermarket. Liquid sam-
ples were appropriately diluted with water and then filtered using
a 0.45 lm pore size filter. Yogurt sample was prepared by adding
10 g of it in 40 ml of deionized water and then stirred for 20 min.
After the sample solution had been centrifuged for 20 min at
1200 g, supernatant solution was filtered in a manner similar to
the liquid samples.
3. Results and discussion
3.1. Comparison of probe solutions
In the indirect UV detection method, a UV-absorbing species
(probe) was used in BGE. When an analyte passed through the
detector, the absorbance decreased and was detected as a negative
peak. Therefore, although no absorbing analytes were present in
the UV and visible ranges, optimal sensitivity could be achieved.
Therefore, it is an effective method considering its ability to simul-
taneously analyse of organic acids and inorganic anions.
The effect of probe species on separation behaviour was exam-
ined using PA, TMA and PMA. Other operating conditions were as
follows: BGE, 200 mM Tris (pH 9.0); signal wavelength, 350 nm;
reference wavelength, 240 nm; capillary temperature, 25 °C; and
applied voltage, 30 kV. Three different types of electrophero-
grams are shown in Fig. 1.
When PA was used as a probe solution, each analyte was de-
tected within nearly 6 min. However, separation of citrate from
tartrate and malate from succinate could not be achieved. When
PMA was applied, the baseline became unstable, and each analyte
demonstrated delayed migration. Although the concentrations of
PA and PMA were changed, the simultaneous analysis of acidulants
and preservatives was difficult. In contrast, when TMA was used,
all acidulants and preservatives were separated successfully. More-
over, the stability of the baseline and the sensitivity of detection
were favourable. However, lower concentrations of TMA resulted
in poor sensitivity, while higher concentrations resulted in the
 K. Yoshikawa et al. / Food Chemistry 127 (2011) 1385–1390 1387

20 5 6
(A) 1~4 7+8
1213
0
9
-20 11

-40 10
3.0 4.0 5.0 6.0 7.0 8.0 9.0

40 34
(B) 12
Absorbance / mAU

5 6
20 7 8 13
12
9
0
11
-20 10
3.0 4.0 5.0 6.0 7.0 8.0 9.0

1
40 2 34
(C)
20 5 6 78
9
0

-20
3.0 4.0 5.0 6.0 7.0 8.0 9.0
Migration time / min

Fig. 1. Comparison of prove species. (A) 10 mM phthalic acid, (B) 10 mM trimellitic acid, (C) 10 mM pyromellitic acid. Analytes (each 50 mg/L): 1. Citrate, 2. Tartrate, 3.
Malate, 4. Succinate, 5. Adipate, 6. Acetate, 7. Propionate, 8. Lactate, 9. Benzoate, 10. Sorbate, 11. Dehydroacetate, 12. Ascorbate, 13. Gluconate; operating conditions: capillary,
bubble cell fused-silica capillary with 50 lm i.d.  64.5 cm (56 cm effective length); BGE, 200 mM Tris adjusted to pH 9.0; injection, 5.0 s at 50 mbar; capillary temperature,
25 °C; applied voltage, 30 kV; signal wavelength, 350 nm; reference wavelength, 230 nm.

disruption of baseline stability. The optimum TMA concentration Although the migration time of each analyte remained largely-
was consequently determined as 10 mM. unaltered, the baseline was entirely unstable at lower Tris concen-
trations in BGE. At Tris concentrations greater than 200 mM, the
3.2. Comparison of indirect UV detection wavelength baseline became similarly unstable within five min of the initiation
of measurement. In addition, the current of BGE was elevated. As a
The effect of the reference wavelength was studied within the result, the optimum concentration of Tris in BGE was determined
range 200–300 nm, with the signal wavelength and band width to be 200 mM primarily on the basis of baseline stability.
fixed at 350 nm and 10 nm, respectively. A solution of 200 mM Tris
and 10 mM TMA (pH 9.0) was used as BGE, and other conditions
3.5. Effect of PVA concentration in BGE
were as described in Section 2.2.
The baseline was unstable at a wavelength of 220 nm or less.
The effect of PVA concentration was examined within the range
Also, the detection sensitivity of benzoate and sorbate was unsat-
0.05–0.15 (w/w)%. The pH of BGE was adjusted to 9.0 and concen-
isfactory because the absorption was rarely different between the
trations of Tris and TMA were fixed at 200 and 10 mM,
species and the probes. In contrast, at a wavelength of 250 nm or
respectively.
more, the detection sensitivity of all analytes became low. Because
Addition of PVA led to easy separation of malate from succinate
the absorption of TMA decreased with increasing wavelength, this
and propionate from lactate. In addition, because EOF was inhib-
was not suitable as a probe for indirect UV detection. As a result,
ited as the viscosity of BGE increased, each analyte showed delayed
the optimum reference wavelength was determined to be
migration. Moreover, the reproducibility of the migration time of
230 nm on the basis of the detection sensitivity of analytes and
each of the analytes was favourable. The optimum concentration
the stability of the baseline.
of PVA in BGE was determined to be 0.10 (w/w)%.

3.3. Effect of pH of BGE on the separation behaviour

The effect of pH of BGE was examined within the range 7.5–9.5,
with the concentrations of Tris and TMA fixed at 200 and 10 mM,
respectively.
At a pH other than 9.0, the baseline was unstable due to the ef-
fect of the pH adjustment solution. In addition, EOF was unstable at
a lower pH. Therefore, the optimal pH of BGE was determined to be
9.0 on the basis of the baseline and EOF stabilities.
3.4. Effect of Tris concentration in BGE
The effect of Tris concentration in BGE was studied within the
range 50–200 mM. The pH was adjusted to 9.0 and TMA concentra-
tion was fixed at 10 mM. Three different types of electrophero-
grams are shown in Fig. 2.
3.6. Other operating conditions
The effects of capillary temperature and applied voltage on the
separation behaviour were examined. BGE was prepared using
200 mM Tris, 10 mM TMA and 0.10 (w/w)% PVA, and the pH was
adjusted to 9.0. The signal wavelength was set at 350 nm with a
reference wavelength at 230 nm. Upon consideration of the resolu-
tions and migration times for each analyte, the optimum capillary
temperature and applied voltage were determined to be 25 °C and
30 kV, respectively. Although the operating conditions (probe
condition, detection wavelength, BGE conditions, capillary temper-
ature and applied voltage) of the acidulants and preservatives were
examined, the established operating conditions in this study were
identical to those described in the published method (Bord et al.,
2005).
1388 K. Yoshikawa et al. / Food Chemistry 127 (2011) 1385–1390

30
(A) 34
20 1 2
6 13
10 5 78 12

0
9 10 11
-10
3.0 4.0 5.0 6.0 7.0 8.0 9.0

30
(B) 12
34
6
Absorbance / mAU

20 5 78
12 13
10
0 9
10 11
-10
3.0 4.0 5.0 6.0 7.0 8.0 9.0

30
(C) 2
34
20 1
5 6
10 78
12 13
0
-10 9
10 11
-20
3.0 4.0 5.0 6.0 7.0 8.0 9.0
Migration time / min

Fig. 2. Effect of Tris concentration in the BGE. (A) 150 mM, (B) 200 mM, (C) 250 mM. Analytes (each 50 mg/L): 1. Citrate, 2. Tartrate, 3. Malate, 4. Succinate, 5. Adipate, 6.
Acetate, 7. Propionate, 8. Lactate, 9. Benzoate, 10. Sorbate, 11. Dehydroacetate, 12. Ascorbate, 13. Gluconate; Operating conditions: capillary, bubble cell fused-silica capillary
with 50 lm i.d.  64.5 cm (56 cm effective length); BGE, 10 mM TMA, 50–250 mM Tris adjusted to pH 9.0; injection, 5.0 s at 50 mbar; capillary temperature, 25 °C; applied
voltage, 30 kV; signal wavelength, 350 nm; reference wavelength, 230 nm.

3.7. Electropherogram of acidulants and preservatives (sucrose, fructose and glucose) present in the food samples were
confirmed. As shown in Table 1, the separation of almost every aci-
A typical electropherogram of nine acidulants and four preser- dulant, preservative and inorganic anion was performed success-
vatives under the optimum operating conditions described in Sec- fully. Although the migration time of fluoride and citrate was
tion 2.3 is shown in Fig. 3. Each species was detected within eight similar, this has little effect on the simultaneous determination
min. Although the separation of propionate and lactate was unsat- of acidulants and preservatives. Because the pH of BGE is lower
isfactory, the data analysis-related quantitative determination was than the dissociation constant of each carbohydrate, a nonionic
applied to peak height calculation. In addition, these species were state exists. Therefore, carbohydrate was not detected in this
simultaneously determined using HPLC and IC. Despite changing method.
the operating conditions, the retention time and separation behav-
iour of these species were unsatisfactory. Therefore, simultaneous
3.9. Analytical precision
analysis of these species by chromatographic technique was found
to be difficult.
Table 2 shows the linearity of the calibration curves, together
with the reproducibility and detection limits. The calibration
3.8. Effect of coexisting species in the food samples curves obtained from the peak heights were plotted with three rep-
licates for each concentration (20, 30, 40, 60, 80 and 100 mg/L). The
The migration times of inorganic anions (bromide, chloride, ni- curves were linear with a correlation coefficient of 0.99. The
trite, nitrate, sulphate, sulphide and fluoride) and carbohydrates relative standard deviations of peak heights with 10 replicates

4
30 3
12

20 5 6
Absorbance / mAU

78
10 13
12
0
9
-10
10 11
-20
3.0 4.0 5.0 6.0 7.0 8.0
Migration time / min

Fig. 3. Electropherogram of acidulants and preservatives. Analytes (each 50 mg/L): 1. Citrate, 2. Tartrate, 3. Malate, 4. Succinate, 5. Adipate, 6. Acetate, 7. Propionate, 8.
Lactate, 9. Benzoate, 10. Sorbate, 11. Dehydroacetate, 12. Ascorbate, 13. Gluconate; Operating conditions: capillary, bubble cell fused-silica capillary with 50 lm i.d.  64.5 cm
(56 cm effective length); BGE, 10 mM TMA, 200 mM Tris and 0.10 (w/w)% PVA adjusted to pH 9.0; injection, 5.0 s at 50 mbar; capillary temperature, 25 °C; applied voltage,
30 kV; signal wavelength, 350 nm; reference wavelength, 230 nm.
 K. Yoshikawa et al. / Food Chemistry 127 (2011) 1385–1390 1389

Table 1 Table 3
Comparison of the migration time. Analytical results of acidulants and preservatives in food sample.
Analytes Migration time/min Organic acid Concentration/mg/L
(I) Acidulants and preservatives White Red wine Health Yogurt Pickle
Citrate 3.768 wine drink of ginger
Tartrate 3.827
Malate 3.892 Citrate 346 ± 15 47 ± 1 3148 ± 51 276 ± 3 58 ± 1
Succinate 3.931 Tartrate 1491 ± 38 1612 ± 19 2136 ± 25 n.d. n.d.
Adipate 4.492 Malate 2196 ± 35 244 ± 4 982 ± 6 n.d. n.d.
Acetate 4.871 Succinate 303 ± 9 491 ± 6 n.d. 19 ± 1 n.d.
Propionate 5.566 Adipate n.d. n.d. n.d. n.d. n.d.
Lactate 5.597 Acetate 190 ± 3 452 ± 8 n.d. 44 ± 2 5752 ± 69
Benzoate 6.198 Propionate n.d. n.d. n.d. n.d. n.d.
Sorbate 6.741 Lactate 334 ± 7 1922 ± 11 n.d. 1739 ± 21 n.d.
Dehydroacetate 7.102 Benzoate n.d. n.d. 897 ± 15 n.d. n.d.
Ascorbate 7.612 Sorbate 178 ± 4 167 ± 9 n.d. n.d. 241 ± 6
Gluconate 7.873 Dehydroacetate n.d. n.d. n.d. n.d. n.d.
Ascorbate 192 ± 9 136 ± 11 n.d. n.d. n.d.
(II) Other species Gliconate 464 ± 8 261 ± 4 n.d. n.d. n.d.
Bromide 2.774
Chloride 2.865 Samples are analysed in triplicate (Mean ± SD).
Nitrite 3.028 n.d., not detected.
Nitrate 3.052
Sulphate 3.244
Sulphite 3.318
Fluoride 3.798
Sucrose n.d. for 20 mg/L were found to range from 1.2% to 4.7%. The detection
Fructose n.d. limits (DLs), summarised in Table 2, were defined as the analyte
Glucose n.d.
concentrations corresponding to a signal equal to three times the
n.d., not detected. background noise (S/N = 3). The DL of each of the analytes was
found to range from 0.6 to 5.3 mg/L.

3.10. Application to food samples

Table 2
Analytical precision of acidulants and preservatives.
The proposed method was applied to analyse the acidulants and
Analytes Linearitya Reproducibilityb Detection limit
preservatives in actual food samples. The analytical results of the
R2 (n = 6) % RSD (n = 10) mg/L (S/N = 3)
acidulants and preservatives in the food samples are shown in
Citrate 0.9985 1.45 0.75
Table 3.
Tartrate 0.9997 1.58 0.75
Malate 0.9998 1.72 0.69 Citrate, tartrate, malate, succinate, acetate, lactate and gluco-
Succinate 0.9996 1.21 0.68 nate were detected as acidulants. Benzoate and sorbate were de-
Adipate 0.9933 3.31 0.78 tected as preservatives. In fact, all species listed in the food
Acetate 0.9906 4.13 0.76 composition table were detected, and the quantitative values were
Propionate 0.9921 1.64 0.84
Lactate 0.9965 2.01 1.02
within the usage standards defined by the Japanese Food Sanita-
Benzoate 0.9958 4.17 2.12 tion Law.
Sorbate 0.9939 1.85 1.04 In addition, to confirm the reliability of the method, 50 mg/L of
Dehydroacetate 0.9921 1.42 0.61 each analyte was added to a food sample, and a recovery test was
Ascorbate 0.9943 4.65 5.25
demonstrated. The results of each recovery rate are shown in Ta-
Gluconate 0.9968 2.49 1.45
ble 4. Because nearly 100% recovery was observed, this method
a
Correlation coefficients for ranged from 20 to 100 mg/L. can be used for the analysis of organic acids, while avoiding the
b
Relative standard deviation at 20 mg/L.
matrix effect.

Table 4
Recovery test of acidulants and preservatives in food sample.

Organic acid White wine Red wine Health drink Yogurt Pickle of ginger
Citrate 104.8 ± 3.8 104.7 ± 3.3 93.9 ± 3.2 95.5 ± 4.1 81.5 ± 0.6
Tartrate 96.1 ± 1.7 94.6 ± 1.3 88.2 ± 1.3 86.0 ± 0.8 90.9 ± 2.1
Malate 97.1 ± 1.4 90.2 ± 1.3 93.4 ± 1.0 89.6 ± 1.7 90.4 ± 1.4
Succinate 99.9 ± 1.2 95.7 ± 0.4 100.4 ± 5.3 98.5 ± 2.0 97.8 ± 0.3
Adipate 101.8 ± 4.1 98.3 ± 1.5 87.5 ± 1.7 91.5 ± 1.8 95.3 ± 0.6
Acetate 106.2 ± 1.7 93.9 ± 1.4 99.1 ± 1.2 96.7 ± 0.8 86.5 ± 5.4
Propionate 106.7 ± 3.1 107.9 ± 1.8 95.9 ± 2.5 105.8 ± 4.3 101.0 ± 1.6
Lactate 86.0 ± 3.5 101.5 ± 4.1 90.5 ± 5.6 106.1 ± 6.3 102.6 ± 3.1
Benzoate 108.5 ± 0.9 101.2 ± 0.6 105.2 ± 5.6 108.8 ± 2.3 102.9 ± 0.8
Sorbate 93.4 ± 0.8 94.9 ± 1.4 102.0 ± 3.8 102.6 ± 1.9 92.2 ± 1.7
Dehydroacetate 102.9 ± 1.0 101.8 ± 0.6 99.9 ± 1.7 100.0 ± 1.8 97.0 ± 2.2
Ascorbate 101.7 ± 5.7 108.2 ± 1.4 87.7 ± 5.7 102.7 ± 1.2 101.3 ± 1.3
Gliconate 104.3 ± 3.0 96.0 ± 0.8 103.1 ± 2.6 101.9 ± 1.9 96.9 ± 2.2

Each analyte (50 mg/L) was added to a food sample.

Samples are analysed in triplicate (Mean ± SD).
1390 K. Yoshikawa et al. / Food Chemistry 127 (2011) 1385–1390
4. Conclusion
A CZE method with indirect UV detection was developed for the
simultaneous analysis of nine acidulants and four preservatives as
defined by the Japanese Food Sanitation Law. EOF is reversed by
using an HDB semi-permanent coated bubble cell fused-silica capil-
lary. When a solution of 10 mM TMA, 200 mM Tris and 0.10 (w/w)%
PVA with the pH adjusted to 9.0 was used as the BGE, each analyte
was detected within 8 min. The linearity of calibration curves and
the reproducibility of peak heights were satisfactory. Although a
high DL was obtained compared with the chromatographic method,
this method was suitable for the analysis of acidulants and preserva-
tives in food samples without the matrix effect. Using this method,
all the species listed in the food composition table were detected,
and the quantitative values were found to be within the usage stan-
dards defined by the Japanese Food Sanitation Law.
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