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Extraction and quantitative determination of ascorbic acid during different

maturity stages of Rosa canina L. Fruit

Article  in  Journal of Food Composition and Analysis · June 2008

DOI: 10.1016/j.jfca.2007.11.007

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 ARTICLE IN PRESS

JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 21 (2008) 300–305
www.elsevier.com/locate/jfca

Original Article

Extraction and quantitative determination of ascorbic acid during

different maturity stages of Rosa canina L. fruit
Saeed Nojavana,c,, Faezeh Khaliliana,b, Fatemeh Momen Kiaiec, Atyeh Rahimic,
Armin Arabanianc, Soheila Chalavia
a
Department of Chemistry, Faculty of Science, Shahid Beheshti University, P.O. Box 19835-389, Tehran, Iran
b
Department of Chemistry, Sharif University of Technology, P.O. Box 11365-9516, Tehran, Iran
c
Department of Quality Control, Tofig Daru Research and Engineering Center, P.O. Box 19395.4978, Tehran, Iran
Received 20 December 2006; received in revised form 31 October 2007; accepted 15 November 2007

Abstract

Dog rose (Rosa canina L.) fruit in different stages of proposed was used as a source of ascorbic acid. Two sample preparation methods
for extracting ascorbic acid in dog rose fruit were evaluated. These methods used high performance of liquid chromatography (HPLC)
for detecting of ascorbic acid, but differed in the preparation of sample (freezing and mild-temperature-drying procedure). Under
optimized conditions, the freezing procedure demonstrated better results. The method was used to compare the amount of ascorbic acid
in fully ripe, half-ripe and unripe dog rose samples. The results show that dog rose has the highest amount of ascorbic acid in its fully ripe
maturity stage. In addition, the intra-day stability of ascorbic acid in standard solution, fully ripe dog rose extract and fully ripe dog rose
intact fruit, was investigated. The results show that ascorbic acid has highest stability in untreated dog rose fruits. As a comparative
study, orange sample was also analyzed by the methodology developed in this work. The results show that the amount of ascorbic acid in
dog rose fruit (417 mg per 100 g) is about 6 times higher than that in orange sample (76 mg per 100 g).
r 2008 Elsevier Inc. All rights reserved.

Keywords: Rosa canina L.; Dog rose; Ascorbic acid; Vitamin C; Freezing; Liquid chromatography; Extraction

1. Introduction the cellular membrane. Vitamin C is important in collagen

Vitamin C belongs to the water-soluble class of vitamins.
Ascorbic acid (AA) is an odorless, white solid having the
chemical formula C6H8O6. This vitamin is easily oxidized
to form dehydroascorbic acid (DHAA), and thus oxidation
is readily reversible from DHAA (Groff et al., 1995).
The importance of vitamin C was first discovered in
1747. It is the major water-soluble antioxidant within the
body. Humans are one of the few species who lack the
enzyme to convert glucose to vitamin C. The vitamin
readily donates electrons to break the chain reaction of
lipid peroxidation. The water-soluble properties of vitamin
C allow for the quenching of free radicals before they reach
Corresponding author at: Department of Chemistry, Faculty of
Science, Shahid Beheshti University, P.O. Box 19835-389, Tehran, Iran.
Tel./fax: +98 21 66026454.
E-mail address: s_nojavan@sbu.ac.ir (S. Nojavan).
formation, thereby resulting in stabilization of the peptide.
Indirectly, AA plays important regulatory roles through-
out the entire body due to its involvement in the synthesis
of hormones, hormone-releasing factors, and neurotrans-
mitters (Groff et al., 1995; Jacoba, 1999).
There is a wide variety of food containing vitamin C.
The general public today generally knows that the best
sources of vitamin C are citrus fruits such as orange and
their juices. A wide variety of other foods also contain
sufficient quantities of vitamin C, such as pineapples, sweet
peppers, broccoli, curly kale, cauliflower, black currants,
and dog rose. Among these foods, dog rose has the highest
amount of vitamin C, so the fruits of these roses were used
to prevent scurvy (Jacoba, 1999).
Rosa canina L., known as dog rose, is a truly multi-
purpose plant that grows wild and can be cultivated in the
garden, and it is very much under-estimated and -utilized.
Teas made from ‘‘rose hips’’ have mild laxative and

0889-1575/$ - see front matter r 2008 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2007.11.007
 ARTICLE IN PRESS
S. Nojavan et al. / Journal of Food Composition and Analysis 21 (2008) 300–305
diuretic tendencies. They help regulate the menstrual cycle,
and stem heavy periods. Infusions made of the leaves and
petals are soothing to the skin, and can help heal rashes
and abrasions. Taken as a tea an infusion of the petals is
good for bringing down fevers, aiding the liver and
gallbladder, and treating the symptoms of colds and
influenza, such as runny noses and sore throats. In
addition, the petals are also good for stopping diarrhea.
Several methods have been developed for the estimation
of ascorbic acid levels in different samples. These include
spectrophotometry (Noroozifar and Khorasani, 2003;
Aydogmus et al., 2002; Sena et al., 2000), calorimetry
(Antonelli et al., 2002), chemiluminescence (Kato et al.,
2005), voltammetry (Ahmed et al., 2005; Ensafi, 2003),
enzymatic assays (Shekhovtsova et al., 2006; Danet et al.,
2000), and amperometric method (Kumar and Narayanan,
2006). However, there may be some drawbacks in
sensitivity, selectivity, stability, or difficulties in sample
preparation. Nowadays, high-performance liquid chroma-
tography (HPLC) (Iglesias et al., 2006; Shakya and
Navarre, 2006; Lopes et al., 2006; Frenich et al., 2005;
Kand’ar et al., 2005; Brause et al., 2003; Yuan and Chen,
1999) and capillary electrophoresis (Law et al., 2005; Wu
et al., 2007; Tang, and Wu, 2005; Zinellu et al., 2004; Jin
and Jiang, 2002), with various detection methods has been
the most used technique for the analysis of ascorbic acid in
different samples. The amount of ascorbic acid has been
reported in dog rose samples using conventional extraction
method (Bozan et al., 1998; Halasova and Jicinska, 1998).
In this work dog rose fruit in different maturity stages
was used as a source of AA. Two methods were applied for
sample preparation and AA content was then determined
in all samples using liquid chromatography method. Also
stability of AA in different type of samples (standard
solution, dog rose extract, and dog rose fruit) was
investigated. In addition, the amount of AA in this fruit
was compared with its amount in orange fruit using the
same sample preparation procedure and chromatographic
method.
2. Methods and materials
2.1. Reagents and chemicals
L-ascorbic acid (AA), metaphosphoric acid (MPA),
orthophosphoric acid, and acetonitrile (HPLC-Grade)
were all purchased from Merck (Darmastdt, Germany).
For chromatographic analysis, de-ionized water of
18 MO cm1 resistivity purified with a milli-Q system
(Millipore, Bedford, USA) was used. Ascorbic acid stock
standard solution was prepared in water and stored in a
glass-stoppered bottle at 4 1C in the dark.
2.2. Plant materials
Dog rose (R. canina L.) fruits were obtained from the
Tehran’s Botanical Garden (Tehran, Iran) in April of 2006.
301
Ripeness stage of dog rose samples was classified by the
color of fruits. So that green, orange, and red colors were
characterized as the sign of unripe, half-ripe and full ripe
samples, respectively. The orange sample was collected at
fully ripeness and used for comparative study.
2.3. Instrumentation
The HPLC system consists of Waters liquid chromato-
graph (Milford, MA, USA) equipped with a 600E multi-
solvent delivery system, an in-line degasser, a manual
injection with 20 mL loop (Rheodyne 7125), and Waters
2487 dual l absorbance detector. Empowers software was
used for controlling the analytical system and data
processing.
2.4. Extraction of ascorbic acid
Sample preparation was performed according to the
freezing method or mild-temperature-drying technique. In
the first method, dog rose samples were sliced, frozen into
liquid nitrogen and in the second method, samples were
dried in mild temperature (15–20 1C) and ground to find
powder dust. These two methods in combination with
extraction processes are described in the following sections.
2.4.1. Freezing procedure
About 100 g of each maturity stages (fully ripe, half-ripe,
and unripe) of dog rose and orange samples were
separately frozen into liquid nitrogen and stored at
70 1C until the analysis were carried out. Frozen
pulverized samples were weighed (1.0 g for orange, 1.0 g
for fully ripe, 1.0 g for half-ripe, and 2.0 g for unripe dog
rose sample) and mixed with 5 mL of the extractant
solution containing 5% of MPA. The mixture was
homogenized for 5 min and then it was centrifuged at
2000 rpm for 10 min. All extractions were carried out under
reduced light and at 4 1C. All the extraction processes was
three times replicated. The extracts were stored at 4 1C less
than 1 h before analysis. Dog rose extracts were diluted 5
times with HPLC-grade water and subsequently were
injected. The injection of extracts into HPLC system was
performed twice.
2.4.2. Mild-temperature-drying procedure
This procedure is a modification of the method done by
Bozan et al. (1998). About 100 g of dog rose samples in
each maturity stage were separately weighed and dried
under mild temperature (15–20 1C) and ground to find
powder dust before extraction. Then obtained powder were
weighed (1.0 g for orange, 1.0 g for fully ripe, 2.0 g half-
ripe, and 2.5 g for unripe dog rose sample) and subse-
quently extracted with 25 mL of extractant solution,
containing 5% MPA, at 10 1C and in the dark. Extraction
process was performed using a shaker for 4 h. All
extractions were carried out in triplicate and obtained
solutions were then filtered and stored at 4 1C before
 ARTICLE IN PRESS
302 S. Nojavan et al. / Journal of Food Composition and Analysis 21 (2008) 300–305
analysis. The injection of the extracts into HPLC system
was performed twice.
2.5. HPLC analysis
The liquid chromatographic method used for the
determination of AA consisted of an isocratic elution
procedure with UV-Visible detection at 245 nm. Separa-
tions were carried out on a 5 mm RP C18 column of
250 mm  4.6 mm (Spherical, Optimals ODS-H, Capital
HPLC, UK) fitted with a 5 mm RP C18 guard column of
20 mm  4.6 mm (Spherical, Optimals ODS-H, Capital
HPLC, UK). The mobile phase employed was a mixture of
0.5% NaH2PO4 (pH 2.25 with H3PO4)–acetonitrile (93:7).
Flow rate of the mobile phase was 1.2 mL min1 and an
injection volume of 20 mL was used in quantitative analysis.
The temperature of analytical column was kept constant at
25 1C. The calibration curve and quantitative evaluations
were accomplished at 245 nm. Standard solutions and
extracts were filtered through a prefilter and then a 0.45 mm
millipore membrane before their injection. To prevent the
loss of AA, standard solutions and extracted samples were
protected from light using amber flasks.
Quantitation was performed by comparing the chroma-
tographic peak area with that of the external standard. The
calibration curve was plotted in the concentration range of
0.5–200 mg L1 and based on a 10-point calibration.
3. Results and discussion
3.1. Optimization of chromatographic conditions
Initially the chromatographic conditions such as flow
rate, mobile phase composition, and column temperature
were optimized. Good results were obtained using a
mixture of orthophosphoric buffer and acetonitrile as the
mobile phase. The concentration of orthophosphoric acid
was studied over the range of 0.1–1% and a concentration
of 0.5% w/w was found to be optimal. The percent of
acetonitrile was studied over the range of 0–15% v/v and a
percent of 7% was found suitable. The best flow rate was
1.20 mL min1 (optimized between 0.7 and 1.5 mL min1).
The oven temperature was studied over the range 20–40
and 25 1C was considered optimum. These parameters were
optimized for time saving while keeping a good resolution
between the peaks of AA and other compound peaks in
the samples. The AA peak authenticity in samples was
confirmed by comparison of an AA peak in samples with
AA in a standard solution. The Waters 2487-dual l
absorbance detector can generate peak purity information
as peaks are being eluted. In the dual wavelength mode, the
detector provides a real time wavelength ratio plot. This
plot is determined by the ratio of the two wavelengths
specified in the method. This ratio also has a threshold
value which can be set so the ratio is plotted only when a
peak is detected; not when the signal is recording baseline.
The ratio will be constant (square waveform) during
detection of an entire pure peak. If the shape of the plot
deviates from the square waveform during detection of
a peak, then the peak can be considered as no longer
spectrally pure.
3.2. Optimization and selection of sample preparation
methods
3.2.1. Freezing procedure
In this method the weight of frozen pulverized sample,
MPA concentration in extractant solution, and centrifuge
time were optimized and the best amounts were obtained
based on extraction recovery. The weight of different
samples were studied over the 0.1–3 g and the suitable
amount for each sample was obtained. The concentration
of MPA was studied and a concentration of 5% was
selected. After homogenization, the time for centrifuge was
studied between 1 and 20 min. From the point of extraction
recovery the best time was 10 min.
3.2.2. Mild-temperature-drying procedure
In this method, concentration of MPA in extractant
solution and time of extraction were optimized. The
concentration of MPA was studied and a concentration
of 5% was selected. The extraction time was studied over
the 0.5–10 h and the 4 h was found optimum. These
parameters were selected from the point of extraction
recovery.
3.3. Measurement of ascorbic acid
3.3.1. Standard solutions
Calibration curve for AA was obtained by plotting the
AA peak area against the AA concentration at 10 levels.
The response of AA over a concentration range of
0.5–200 mg L1 was linear (Y ¼ 60.384x+1.437) with a
regression coefficient (R2) of 0.997.
The limit of quantitation (LOQ) and detection (LOD)
were 0.20 and 0.05 mg L1, respectively, which was taken
as the amount of ascorbic acid giving a signal-to-noise ratio
greater than 3. The RSD values for repeatability and intra-
day reproducibility (n ¼ 6) for a standard solution contain-
ing 25 mg L1 of AA were 1.24% and 7.20%, respectively.
For study of intra-day reproducibility the standard
solution was kept at 4 1C and in the dark.
3.3.2. Quantification of AA in samples
Table 1 shows AA content for the fully ripe, half-ripe,
and unripe dog rose samples based on described sample
preparation methods. According to the results, the
amounts of AA varied greatly at three maturity stages of
dog rose samples. Comparing two sample preparation
methods is also demonstrated that there is a substantial
difference between these two methods. The AA content
were obtained 1871.2 (unripe dog rose), 17573.7 (half-
ripe dog rose), and 417.573.2 (fully ripe dog rose) mg per
100 g1 fresh weight for the freezing method, whereas for
 ARTICLE IN PRESS
S. Nojavan et al. / Journal of Food Composition and Analysis 21 (2008) 300–305 303

the mild-temperature-drying sample preparation method
the concentrations were 370.4 (unripe dog rose), 3472.7
(half-ripe dog rose), and 21174.3 (fully ripe dog rose) mg
per 100 g1 fresh weight. None of the extracts contained
interfering substances. Fig. 1 shows the chromatograms for
fully ripe and unripe dog rose extracts that obtained by
freezing sample preparation procedure. The obtained
chromatograms reveal the absence of interfering sub-
stances. The ascorbic acid amount found in this study for
fully ripe dog rose was higher than that obtained by Bozan
for dog rose (48–114.3 mg per 100 g), while this amount for
half-ripe dog rose was similar to those found by Bozan
et al. (1998). The method proposed by Bozan et al. was
applied for dog rose sample. Since the obtained recoveries
for the target samples were much less than the reported
values, this method was modified in a way that an
Table 1
Comparison of ascorbic acid content obtained with two sample
preparation methods in three maturity stage of dog rose samples
Sample Ascorbic acid mg per 100 g
Freezing
method
Full-ripe dog rose 417.573.2
Half-ripe dog rose 17573.7
Unripe dog rose 1871.2
appropriate compromise between mild extraction tempera-
ture and time was obtained. The difference in type of
samples, grinding of samples, maturity of samples, and
other such conditions can cause different results between
our laboratory and the Bozan laboratory.
In order to evaluate the matrix effect on the accuracy of
analysis, a recovery test was carried out. AA was added to
dog rose samples at two different concentration levels
(200 and 500 mg of AA) and analyzed in triplicate using the
two sample preparation methods. The results are shown in
Table 2. The mean recovery for AA with freezing sample
preparation method was 10074% and the RSD for AA
ranged from 2.42% to 6.24%. Due to the low recovery
(mean recovery 57.374.1%) of the mild-temperature-
drying method, determination of AA in all of maturity
stages of dog rose is inconvenience. Putting all the data
together indicates that the freezing method is more suitable
for quantitative analysis of AA in dog rose and other
similar samples. It is well known that ascorbic acid is a
sensitive and thermo-labile compound, so its extraction
with high yield and without any decomposition has been a
matter of difficulty and labor-intensive for the analysts. In
this work it has been tried to extract ascorbic acid in a way
Mild-temperature
drying method that the obtained recoveries will be as much as possible.
In comparison with the modified method reported by
21174.3 Bozan et al. (1998), extraction of ascorbic acid using
3472.7
freezing protocol provides more reliable results in terms
370.4
of recovery and reproducibility. The repeatability of the

1180 75
AA
980 AA 65
55
780
Volt (mV)

Volt (mV)

45
580 35
380 25
15
180 5
-20 -5
0 2 4 6 8 10 0 1 2 3 4 5 6 7 8 9
Time (min) Time (min)

Fig. 1. Typical Chromatograms obtained at 245 nm for (A) fully ripe and (B) unripe dog rose. Samples prepared using freezing procedure. Optimized
HPLC condition: flow rate 1.2 mL min1, isocratic, mobile phase was a mixture of phosphate buffer (0.5%)–acetonitril (93:7). Ascorbic acid retention time
(6.15 min).

Table 2
Ascorbic acid recovery test of freezing versus mild-temperature drying method

Fruit maturity stage Freezing method Mild temperature drying method

200 mg added 500 mg added 200 mg added 500 mg added

Ra (%) RSD (%) R (%) RSD (%) R (%) RSD (%) R (%) RSD (%)

Unripe 9874 4.41 10272 2.42 5373 3.32 6776 5.65

Half-ripe 10076 5.75 9776 6.24 4877 6.64 5773 3.41
Full-ripe 9973 3.23 10175 4.73 5774 3.72 6272 1.87
a
R: Recovery.

304 S. Nojavan et al. / Journal of Food Composition and Analysis 21 (2008) 300–305
Table 3
The percentage of ascorbic acid calculated based on the ratio of AA
content after definite time versus fresh sample
Sample The ratio of ascorbic acid content to
fresh solution %
1st day 5th day
Standard solutiona 98 68
Spiked sampleb 99 73
Ripe dog rose plant 101 94
a 1
25 mg mL AA in 5% MPA.
b
Full-ripe dog rose extract spiked with 25 mg L1 AA.
extracts ranged between 3% (fully ripe dog rose) and
8.7% (unripe dog rose) and the RSD values for reprodu-
cibility between 4.2% and 9.2% for fully ripe and unripe
dog rose, respectively. This results were shown that
there is no significant differences between the within day
and intra-day precision and the freezing method has good
reproducibility.
3.4. Intra-day stability of AA
The effect of storage time (1–15 day) on the stability of
AA was investigated at room temperature. This study was
performed using a 25 mg L1 AA standard solution in 5%
MPA, a fully ripe dog rose extract spiked with 25 mg L1
AA, and fully ripe dog rose intact fruit that left at room
temperature. Table 3 shows these results. After 1 day, the
stability of AA kept at room temperature was 9972%,
which was similar to fresh solution. After 5 days, it was
6873%, 7372%, and 9472% and after 15 days, it was
1772%, 3671%, and 8773% for standard solution,
spiked dog rose extract, and dog rose fruit, respectively.
These results demonstrated so that the stability of ascorbic
acid in fruit matrix is higher than the fruit extract.
Percentages are calculated based on the ratio of AA
content after definite time versus fresh sample.
3.5. Comparison of dog rose with orange
Since orange is a good source of AA. For this reason the
amount of AA in dog rose was compared with orange.
Orange extract was analyzed in the same LC condition.
Fig. 2 shows typical chromatogram of an orange sample.
As can be seen, the retention time is in agreement with that
of AA standard peak, and there is a good resolution. The
AA content in orange sample was obtained 7672.1 mg per
100 g fresh weight using the freezing method. The obtained
results show that the amount of AA in dog rose fruit is
about six times more than that in the orange sample.
4. Conclusions
It is well known that ascorbic acid is a sensitive and
thermo-labile compound, so its extraction with high yield
ARTICLE IN PRESS
150
AA
130
110
90
Volt (mV)
70
50
15th day
30
17 10
36
-10
87 0 2 4 6 8 10
Time (min)
Fig. 2. Chromatogram of orange sample, obtained at the same condition
of Fig. 1. Sample was prepared using the freezing procedure.
and without any decomposition has been a matter of
difficulty and labor-intensive for the analysts. The freezing
sample preparation method prevents oxidation and degra-
dation of ascorbic acid and affords enough sensitivity
and selectivity in extraction of ascorbic acid in dog rose
(at three maturity stages) and orange sample. Also the
proposed LC method has good resolution from other
contaminants or the solvent peak. The method delivers
results within 6.1 min after fruit extraction. The proposed
method can be used to identify and quantify vitamin C in a
wide variety of fruits. Finally, the stability of ascorbic acid
in the presence of fruit matrix is higher than the fruit
extracts at room temperature.
Acknowledgment
We gratefully acknowledge Dr. Shahryar Ommidvar
Corporation for supporting this project in Tofig Daru
Research and Engineering Center.
References
Ahmed, R., Qureshi, S.A., Viqar, U.N., 2005. Studies about the
determination of ascorbic acid by differential pulse voltammetry.
J. Chem. Soc. Pakistan 27, 168–173.
Antonelli, M.L., D’Ascenzo, D., Laganà, A., Pusceddu, P., 2002. Food
analyses: a new calorimetric method for ascorbic acid (vitamin C)
determination. Talanta 58, 961–967.
Aydogmus, A., Cetin, S.M., Ozgur, M.U., 2002. Determination of
ascorbic acid in vegetables by derivative spectrophotometry. Turk.
J. Chem. 26, 697–704.
Bozan, B., Sagdullaev, B.T., Kozar, M., Aripov, K.H.N., Baser, K.H.C.,
1998. Comparison of ascorbic and citric acid contents in Rosa canina
L. fruits growing in Central Asian region. Chem. Nat. Compd. 34,
687–689.
Brause, A.R., Woollard, D.C., Indyk, R.E., 2003. Determination of total
vitamin C in fruit juices and related products by liquid chromato-
graphy: interlaboratory study. J. AOAC Int. 86, 367–374.
Danet, A.F., Badea, M., Aboul-Enein, H.Y., 2000. Flow injection system
with chemiluminometric detection for enzymatic determination of
ascorbic acid. Luminescence 15, 305–309.
Ensafi, A.A., 2003. Determination of ascorbic acid by electrocatalytic
voltammetry with methylene blue. Anal. Lett. 36, 591–604.
Frenich, A.G., Torres, M.E.H., Vega, A.B., Vidal, J.L.M., Bolanos, P.P.,
2005. Determination of ascorbic acid and carotenoids in food
 ARTICLE IN PRESS
S. Nojavan et al. / Journal of Food Composition and Analysis 21 (2008) 300–305
commodities by liquid chromatography with mass spectrometry
detection. J. Agric. Food Chem. 53, 7371–7376.
Groff, J.L., Gropper, S.S., Hunt, S.M., 1995. The water soluble vitamins.
In: Advanced Nutrition and Human Metabolism. West Publishing
Company, Minneapolis, pp. 222–231.
Halasova, J., Jicinska, D., 1998. Amount of ascorbic acid in the hips of
Rosa species. Folia Geobot 23, 181–185.
Iglesias, J., Gonzalez, M.J., Medina, I., 2006. Determination of ascorbic
and dehydroascorbic acid in lean and fatty fish species by high-
performance liquid chromatography with fluorometric detection. Eur.
Food Res. Technol. 223, 781–786.
Jacoba, R.A., 1999. Vitamin C. In: Modern Nutrition in Health and
Disease, 9th. Williams & Wilkins, Baltimore, pp. 467–473.
Jin, W.R., Jiang, L., 2002. Measurement of ascorbic acid in single human
neutrophils by capillary zone electrophoresis with electrochemical
detection. Electrophoresis 23, 2471–2476.
Kand’ar, R., Zakova, P., Richter, M., 2005. Determination of ascorbic
acid in human plasma by high-performance liquid chromatography
with ultraviolet detection with a view to stability. Clin. Chim. Acta
355, S220–S221.
Kato, T., Ohno, O., Nagoshi, T., Ichinose, Y., Igarashi, S., 2005.
Determination of small amounts of L-ascorbic acid using the
chemiluminescence of an iron-chlorophyllin complex. Anal. Sci. 21,
579–581.
Kumar, S.S., Narayanan, S.S., 2006. Amperometric sensor for the
determination of ascorbic acid based on cobalt hexacyanoferrate
modified electrode fabricated through a new route. Chem. Pharm.
Bull. 54, 963–967.
Law, W.S., Kuban, P., Zha, J.H., Li, S.F.Y., Hauser, P.C., 2005.
Determination of vitamin C and preservatives in beverages by
conventional capillary electrophoresis and microchip electrophoresis
View publication stats
305
with capacitively coupled contactless conductivity detection. Electro-
phoresis 26, 4648–4655.
Lopes, P., Drinkine, J., Saucier, C., Glories, Y., 2006. Determination
of L-ascorbic acid in wines by direct injection liquid chromatography
using a polymeric column. Anal. Chim. Acta 555, 242–245.
Noroozifar, M., Khorasani, M.M., 2003. Application of potassium
chromate-diphenylcarbazide in the quantitative determination of
ascorbic acid by spectrophotometry. Turk. J. Chem. 27, 717–722.
Sena, M.M., Fernandes, J.C.B., Rover, L., Poppi, R.J., Kubota, L.T.,
2000. Application of two- and three-way chemometric methods in the
study of acetylsalicylic. Anal. Chim. Acta 409, 159–170.
Shakya, R., Navarre, D.A., 2006. Rapid screening of ascorbic acid,
glycoalkaloids, and phenolics in potato using high-performance liquid
chromatography. J. Agric. Food Chem. 54, 5253–5260.
Shekhovtsova, T.N., Muginova, S.V., Luchinina, J.A., Galimova, A.Z.,
2006. Enzymatic methods in food analysis: determination of ascorbic
acid. Anal. Chim. Acta 573, 125–132.
Tang, Y.J., Wu, M.J., 2005. A quick method for the simultaneous
determination of ascorbic acid and sorbic acid in fruit juices by
capillary zone electrophoresis. Talanta 65, 794–798.
Wu, T., Guan, Y.Q., Ye, J.N., 2007. Determination of flavonoids
and ascorbic acid in grapefruit peel and juice by capillary
electrophoresis with electrochemical detection. Food Chem. 100,
1573–1579.
Yuan, J.P., Chen, F., 1999. Simultaneous separation and determination of
sugars, ascorbic acid and furanic compounds by HPLC-dual detection.
Food Chem. 64 (3), 423–427.
Zinellu, A., Carru, C., Sotgia, S., Deiana, L., 2004. Optimization of
ascorbic and uric acid separation in human plasma by free zone
capillary electrophoresis ultraviolet detection. Anal. Biochem. 330,
298–305.