ORI GI NAL PAPER

A simplied spectrophotometric method for the determination

of inulin in Jerusalem artichoke (Helianthus tuberosus L.) tubers
Araya Saengkanuk

Suporn Nuchadomrong

Sanun Jogloy

Aran Patanothai

Supalax Srijaranai
Received: 1 April 2011 / Revised: 5 July 2011 / Accepted: 23 July 2011 / Published online: 10 August 2011
Springer-Verlag 2011
Abstract A simple spectrophotometric method was
developed for the analysis of inulin in Jerusalem artichoke
tubers. The inulin was extracted from the artichoke tuber
samples using accelerated solvent extraction method, before
subsequent hydrolysis in acid condition. The hydrolysates
were then analyzed for fructose using spectrophotometry.
The spectrophotometric method is based on oxidation of
fructose by periodate and evaluation of the remaining per-
iodate by measuring the absorbance at 350 nm of the tri-
iodide complex formed, upon addition of potassium iodide.
The optimum conditions for the detection of fructose were
0.1 mmol L
-1
periodate and 1.5 mmol L
-1
potassium
iodide at pH 6.0. The proposed method was validated for its
analytical performance parameters including accuracy,
precision, and recovery. The method was applied to the
determination of inulin in ten varieties of Jerusalem arti-
choke grown in the northeastern part of Thailand. The inulin
content in the samples was found to be in the range of
6375.5% dry weight, and the degree of polymerization was
in the range of 1420. The inulin contents obtained from the
proposed spectrophotometry were not signicantly different
(p = 0.05) from those obtained from high-performance
anion-exchange chromatography coupled with pulsed
amperometric detection. The results indicated that the
present spectrophotometric method can be used as an alter-
native to chromatographic analysis for the determination of
inulin in plant samples.
Keywords Inulin Jerusalem artichoke Fructose
Spectrophotometry HPAEC-PAD
Introduction
Inulin is a linear polymer of D-fructose joined by b (2 ? 1)
linkages and terminated with a D-glucose, which is linked
to fructose by an a(1 ? 2) bond [1, 2]. Inulin has a degree
of polymerization (DP) in the range of 260 [3, 4]. It is a
natural carbohydrate source found mainly in roots and
tubers of many plants, such as chicory artichoke (Cynara
scolymnus L.), vipers grass (Scorzonera hispanica L.), and
Jerusalem artichoke (Helianthus tuberosus L.) [57], which
has been reported to have an inulin content of 1419% (wet
weight). Nutritionally, inulin and its derived fructooligo-
saccharides (FOS) can stimulate health-promoting gut
microora, relieve constipation, improve calcium avail-
ability, and may reduce the risk of cancer [5, 8]. More
recently, inulin has been described as a so-called prebiotic
agent or food supplement not digested nor reabsorbed in
the gastrointestinal system [6, 9, 10]. It has also been used
as a soluble ber macronutrient substitute, especially in the
recipe of dietetic foods for diabetic patients. Various
applications of inulin require different degrees of inulin
quality, which is determined by its chain length or the
degree of polymerization, or DP. Inulin with a low DP is
appropriate for alcohol fermentation, or for production of
FOS, while inulin with a high DP is valuable for
A. Saengkanuk S. Srijaranai (&)
Department of Chemistry and Center for Innovation in
Chemistry, Faculty of Science,
Khon Kaen University, Khon Kaen 40002, Thailand
e-mail: supalax@kku.ac.th
S. Nuchadomrong
Department of Biochemistry, Faculty of Science,
Khon Kaen University, Khon Kaen 40002, Thailand
S. Jogloy A. Patanothai
Department of Plant Science and Agricultural Resources,
Faculty of Agriculture, Khon Kaen University,
Khon Kaen 40002, Thailand
1 3
Eur Food Res Technol (2011) 233:609616
DOI 10.1007/s00217-011-1552-3
pharmaceutical [11]. With regard to its nutritional appli-
cations, the development of analytical methods for
extraction, quantication, and determination of DP is of
great importance for the characterization of inulin and the
derivatives in plant samples.
Determination of inulin can be performed using either
the direct or indirect approach. High-performance anion-
exchange chromatography with pulsed amperometric
detection (HPAEC-PAD) has been accepted as the most
powerful method for direct determination of inulin [1216].
It provides not only the content of inulin but also the DP
proles. However, the difculty in interpreting the elution
order of sugar oligomers was reported due to the lack of
availability of suitable standards. Moreover, the analytical
anion exchange columns are of relatively high cost. Indirect
determination methods are based on hydrolysis of inulin
followed by measurement of the released fructose and
glucose by different techniques including HPAEC-PAD [3,
4, 8, 9], as well as spectrophotometry using various reagents
for derivatization such as dinitrosalicylic acid (DNS) [17]
and p-hydroxybenzoic acid hydrazide (PAHBAH) [18, 19].
Many reports using analytical methods based on enzymatic
hydrolysis and detection have been published [9, 20, 21].
However, the procedures involve several types of expensive
enzymes and/or time-consuming reactions. Therefore,
development of a simple analytical method using common
chemicals available in laboratories for the determination of
inulin is of interest. A simple reaction using iodine has been
used for the determination of sugars [22], and a recent
report [23] describes the use of spectrophotometry based on
the periodate reagent to measure the amount of glucose and
fructose in syrup samples. We describe here, for the rst
time, the application of this simple method to the analysis of
inulin in Jerusalem artichoke tuber samples using acceler-
ated solvent extraction (ASE), followed by hydrolysis and
analysis by the periodate-based spectrophotometry. ASE,
also known as pressurized liquid extraction (PLE), has been
accepted as a powerful extraction, and it performs under
high pressure, thus the extraction can be achieved in short
time [24]. The parameters affecting the extraction, hydro-
lysis, and subsequent chemical reactions involved in the
spectrophotometry were also optimized. Inulin contents
obtained from the proposed spectrophotometric method was
compared to those from HPAEC-PAD. Moreover, the DP
values were calculated using the results from HPAEC-PAD.
Materials and methods
Chemicals and reagents
All chemicals and reagents were of analytical reagent
grade. These included sodium hydroxide, potassium iodide,
and sodium periodate (Carlo Erba, Rodano, Italy); phe-
nolphthalein (Fluka, Buchs, Switzerland); methanol (Lab-
Scan, Bangkok, Thailand); perchloric acid (Merck,
Darmstadt, Germany); silica gel (0.06 mm for chroma-
tography), oxalic acid, absolute ethyl alcohol, and barium
acetate (Carlo Erba, Val de Reuil, France); inulinase from
Aspergillus niger (Sigma, Postal, Denmark), which is a
mixture of exo- and endo-inulinase was used. Standard
chicory inulin (Sigma, Berlin, Germany); standard Beneo
HP, which is chicory inulin, having long fructosyl chains
with average DP of 25 (Orafti, Tienen, Belgium); sodium
tetraborate decahydrate and boric acid (Fluka, Steinheim,
Germany); citric acid, concentrated hydrochloric acid, and
concentrated sulfuric acid (Carlo Erba, Val de Reuil,
France); monosaccharides including D-(-)-fructose and
D-(?)-glucose anhydrous (Univar, New South Wales,
Australia). All aqueous solutions were prepared in deion-
ized water from RiO
s
TM
Type I Simplicity 185 (Millipore,
USA) with the resistivity of 18.2 MX cm.
Instrumentation
The spectrophotometric experiments were carried out on an
Agilent 8453 UVVis spectrophotometer (Germany).
Chromatographic separations were performed on a Dionex,
DX-500 Ion Chromatographic system (Sunnyvale, USA),
consisting of a GP40 gradient pump and a ED 40 electro-
chemical detector equipped with a thin-layer type amper-
ometric cell. The cell contained a 1.0-mm diameter gold
working electrode and a platinum counter electrode in
integrated PAD mode. The separations were performed on
a CarboPac PA10 column set comprised of a guard column
(50 9 2 mm i.d.) and an analytical column (250 9 2 mm
i.d.). The injection volume was 25 lL. The column was
placed inside a temperature controlled unit. The control of
the system, data acquisition, and data analysis were under
the control of the PeakNet 6.0 Dionex software. The
accelerated solvent extractor model ASE-200 (Dionex,
USA), with a 11-mL extraction cell and a 30-mL collection
cell, was used for the extraction of artichoke samples.
Sample preparation
Eleven varieties of Jerusalem artichoke (H. tuberosus L.)
were selected, including CN52867, KKU AC001, HEL 61,
HEL65, HEL66, HEL69, HEL231, HEL335, JA37, JA38,
and JA 102. These were planted on July 2008 at the Field
Crop Research Station of Khon Kaen University, located in
Khon Kaen Province, Thailand (1628
0
N, 10248
0
E, 200 m
above sea level), and harvested on October 2008. The rep-
resentative samples used for the optimization and method
validation experiments are Beneo HP and Jerusalem arti-
choke (CN 52867). Tubers of ve sampled plants for each
610 Eur Food Res Technol (2011) 233:609616
1 3
variety were randomly collected and washed with tap water
to eliminate soil. The tubers were longitudinally sliced at the
middle to get approximately 2-mm thick pieces. Fifty grams
(50 g) of tuber slices (5 sampling plants 9 50 g = 250 g in
total sample weight) were soaked in absolute ethyl alcohol
for 24 h at 4 C. The samples were nally stored at -20 C
in plastic zip-closure bags. Before extraction by ASE, the
samples were dried at 60 C for 10 h in an oven, milled and
sieved through an 850-lm sieve. The powdered samples
were kept for short periods of time at an ambient atmosphere
in desiccators, until ASE extraction.
Extraction of inulin from artichoke samples by ASE
A cellulose lter (P/N 049458) was placed at the outlet of
the extraction cell before loading accurately weighed
samples (2.0 g) along with silica gel (3.6 g), and the top of
the cell was subsequently covered with two Whatman No.
42 lter papers. The samples were then automatically
extracted with water (as the extraction solvent) for 20 min
at 80 C and 1,500 Psi, controlled by the ASE time
program.
Analysis of free fructose content (F
f
)
The reaction was conducted by mixing an aliquot of extracts
(150 lL) with 10 mmol L
-1
sodium periodate reagent
(100 lL), 20 mmol L
-1
citrate buffer pH 6.0 (5.00 mL),
and water (4.60 mL). After 5 min, 100 mmol L
-1
potas-
sium iodide (150 lL) was added, and the mixture was left
for an additional 5 min. The solution absorbance was sub-
sequently measured at 350 nm using a UVVis spectro-
photometer. The concentration of free fructose was deduced
from calibration curve of standard fructose.
Determination of total fructose content (F
tot
)
The extract (1.00 mL) was acidied with 0.2 mol L
-1
HCl,
in a nal volume of 50 mL, and subjected to acid hydro-
lysis at 97 2 C for 45 min. The solution was then
adjusted to pH 7.0 with NaOH before diluting with water to
50.00 mL. The neutral hydrolysate (150 lL) was analyzed
spectrophotometrically by the same procedure as described
above for free fructose (F
f
) analysis.
HPAEC-PAD analysis conditions
The neutral hydrolysate (100 lL) was diluted to 10.00 mL
with water and ltered through a 0.45-lm membrane lter,
before analysis by HPAEC-PAD. The separation conditions
were 10 mmol L
-1
NaOH, spiked with 0.55 mmol L
-1
barium acetate, as the eluent, a ow rate of 1.0 mL min
-1
,
and the CarboPac PA 10 analytical column equilibrated at
30 C. The electrical potential and time period (waveform)
for the integrated PAD were programmed as follows: E1,
?0.05 V (vs. Ag/AgCl) at 0 s (duration step); E2, ?0.05 V
at 0.200.40 s (integration step); E3, ?0.75 V at 0.41
0.60 s, and E4, -0.15 V at 0.611.00 s (cleaning step).
Results and discussion
Optimum conditions for the extraction of inulin by ASE
The extraction of inulin from plant samples before analysis
is an important step in obtaining accurate results to assess
the quality of inulin. Typically, plant samples have been
extracted with hot water ([80 C) for 1 h or longer [4, 25].
Ethanol has also been used as the extraction solvent with
good recoveries [6, 15, 26]. These strategies are effective
for inulin extraction, but are quite time consuming and use
large amounts of solvent for the overall extraction process.
To avoid these problems, accelerated solvent extraction
(ASE) was adopted in this study to extract inulin from
Jerusalem artichoke samples. The parameters under study
included extraction temperature (70100 C) and extrac-
tion time (1030 min). It was found (data not shown) that
the highest extraction yield of fructose and glucose (93%
on average, as determined from HPAEC-PAD), were
obtained by heating at 80 C for 20 min. Longer extraction
times, along with higher temperatures, resulted in
decreased inulin recovery. It may be due to acid-heat
sensitivity of fructose molecule [27]. In addition, the con-
ventional extraction was also performed at its optimum
condition (85 C for 1 h). The results demonstrated the
effectiveness of the simple automated ASE method. The
extraction time could be reduced by at least 3 times in
comparison to the regular method described in previously
published reports.
Hydrolysis condition of inulin
Acid hydrolysis is a powerful and simple method for
hydrolyzing inulin to release glucose and fructose com-
ponents, and it is also much less expensive than the
enzymatic hydrolysis [28]. In our experiments, the acid
hydrolyses were performed at the xed time of 15 min and
at the incubation temperature of 97 C. The effects of three
acid types (perchloric acid, hydrochloric acid, and oxalic
acid, each at the concentration of 1%) were investigated.
The released glucose and fructose were separated and
quantied using HPAEC-PAD. Typical chromatograms
obtained in hydrolysis experiments with different acid
types are shown in Fig. 1, demonstrating that hydrochloric
acid (HCl) hydrolysis gave the highest HPAEC-PAD
Eur Food Res Technol (2011) 233:609616 611
1 3
detection responses for fructose and glucose. It can be seen
that peaks other than fructose and glucose, referred as
by-product peaks, were present in the chromatograms.
However, HCl hydrolysis produced fewer by-product
compounds compared to perchloric and oxalic acids.
Hydrochloric acid was reported as the most efcient acid
for inulin hydrolysis [27, 2931]. Subsequent experiments
addressed the hydrolysis at various concentrations of HCl
in the range 0.10.8 mol L
-1
. Concentrations of HCl
higher than 0.4 mol L
-1
were shown to cause a signicant
decrease in the fructose yield (Fig. 2), while the by-product
compounds were more apparent as the acid concentration
increased (data not shown). It has been reported that
extreme conditions (i.e., low pH, high temperature) may
decrease fructose content, since fructose is more easily
decomposed at high temperatures and in acidic medium
than glucose [27].
The effect of incubation times (1560 min) in hydro-
lysis by 0.2 mol L
-1
HCl was studied (Fig. 3). It was
observed that detectable fructose, the main inulin constit-
uent, and glucose in the inulin hydrolysate increased with
longer incubation times. However, 45 min resulted in the
highest fructose content. More glucose was released at
60-min incubation, and fructose decreased. Incubation for
the longer time, even at lower acidity (0.1 mol L
-1
HCl)
produced undesirable by-products in the system (Fig. 3).
Therefore, 45-min incubation was selected as a compro-
mise for the production of both target compounds and
minimization of undesired compounds during the hydro-
lysis reaction. Higher concentration of HCl and longer
hydrolysis time resulted in decreased fructose content,
which may due to the acid-heat sensitivity of fructose.
Moreover, by-products were observed at longer hydrolysis
time, which was previously reported [31].
To summarize, the optimum conditions for acid hydro-
lysis were 0.2 mol L
-1
HCl and 45-min incubation at
97 C. Under these conditions, the Beneo HP inulin stan-
dard (DP = 25, on average) at 200 mg L
-1
was investi-
gated. The contents of glucose and fructose were 8.04 and
205.59 mg L
-1
, respectively. The recovery of the method
was tested to evaluate the accuracy by spiking glucose and
fructose standards (12 mg L
-1
each) into the standard
Beneo HP, before hydrolysis. The recoveries of fructose
and glucose after their acid hydrolysis and analysis by
HPAEC-PAD were 9496% (n = 3). The chromatograms
of chicory inulin and Beneo HP inulin standards after acid
hydrolysis are illustrated in Fig. 4.
Moreover, enzymatic hydrolysis was also investigated
by the modied method of Lopez [5], using inulinase
produced from Aspergillus niger. A 2.00 mL solution
containing 2 mg inulin was made up to 10.00 mL with
Fig. 1 Chromatograms of standard inulin (200 mg L
-1
) after acid
hydrolysis with a perchloric acid (1%), b hydrochloric acid (1%), and
c oxalic acid (1%). Peak assignment: G glucose; F fructose
Fig. 2 Effect of HCl concentration on the fructose and glucose
production after hydrolysis
Fig. 3 Effect of hydrolysis reaction time on the produced fructose
and glucose
612 Eur Food Res Technol (2011) 233:609616
1 3
50 mmol L
-1
citrate buffer pH 4.8 in the presence or
absence of 1.00 mL inulinase. The mixture was incubated
at 37 C for 12 h. The clear supernatant was subjected for
further analysis by HPAEC-PAD. The chromatograms
obtained from different enzymatic hydrolysis conditions
are shown in Fig. 5. There are many interfering peaks near
fructose and glucose, which may due to incomplete
hydrolysis under the conditions used. However, the com-
plete hydrolysis may occur at longer incubation time.
Optimization for spectrophotometry using periodate
reagent
The proposed spectrophotometric method is based on the
oxidation of glucose and fructose by periodate (IO
4
-
) and
evaluation of the remaining periodate by monitoring the tri-
iodide (I
3
-
) complex formed after the addition of potas-
sium iodide (I
-
). The proposed reaction is depicted in the
following equations:
IO

4
C
6
H
12
O
6
sugars ! Oxidation products IO

4
1
IO

4
7I

8H

! 4I
2
4H
2
O 2
I
2
I

! I

3
brown color; k
max
350 nm 3
The optimum conditions for the proposed
spectrophotometric method were determined by varying
the solution pH, reaction time of periodate and sugars,
reaction time for generated I
3
-
, and concentration of
reagents (periodate reagent and potassium iodide).
Effect of solution pH
The effect of pH was studied over the range of 2.011.0
(data not shown). It was found that at pH 6.0, fructose
reacted to produce the desired color, and at pH 9.0, both
glucose and fructose reacted. Thus, concentrations of each
sugar could be calculated by carrying out the reaction at
two different pHs.
Effect of reaction time for the developed reactions
Since the reactions in this study are stepwise reactions, the
optimum reaction time for each step was investigated and
the results are depicted in Fig. 6a. The absorbance differ-
ence between blank reagent solution and mixture of both
sugars (lower two lines) was suitable for monitoring the
target species after 5 min of reaction, and signals were
almost constant afterwards. Meanwhile, the determination
of fructose can be performed immediately after reaction.
The reaction time of 5 min was chosen as a compromise
between absorbance signal strength and time required for
the analysis of fructose and mixture of sugars.
The optimal reaction time for the iodide reaction (Eq.
#3) was also studied. As shown in Fig. 6b, a sharp increase
in absorbance for fructose analysis was observed during the
rst 5 min of the reaction. After that, the signal was almost
constant. For the mixture of both sugar analytes, varying
the reaction time from 1 to 13 min led to a slight increase
in absorbance signal over this range. Hence, the reaction
time of 5 min was chosen as the optimum.
Effect of concentrations of sodium periodate and potassium
iodide
Because this analytical method measures excess periodate
after conversion to tri-iodide, it is important to keep the
concentration of the periodate reagent high enough to fully
Fig. 4 Chromatograms of standard inulin from a Chicory and from
b Beneo HP, after acid hydrolysis under the optimum condition
Fig. 5 The chromatograms of inulin obtained from enzymatic hydro-
lysis. Assignments: a blank enzyme (2 mL); b enzyme (2 mL) + chic-
ory standard inulin (200 mg L
-1
); c enzyme (1 mL) + chicory
standard inulin (200 mg L
-1
); d enzyme (2 mL) + Beneo HP inulin
standard (200 mg L
-1
)
Eur Food Res Technol (2011) 233:609616 613
1 3
oxidize the sugars, but low enough to keep the measured
tri-iodide in a reasonable range for spectrophotometric
analysis (about 1.0 absorbance unit). The optimum condi-
tions for the determination of fructose are sodium periodate
0.10 mmol L
-1
and potassium iodide 1.50 mmol L
-1
, at
pH 6.0, while, the conditions for both glucose and fructose
(1:1 mixture) determination are sodium periodate
0.20 mmol L
-1
and potassium iodide 2.50 mmol L
-1
, at
pH 9.0. Therefore, glucose can be quantied by the dif-
ference in the contents of mixture and fructose.
Analytical characteristics and validations
of the proposed method
Periodate ion reacts with sugars, which have vicinal alde-
hyde groups. However, the reaction rate depends on
experimental parameters of pH, time, and concentration
[25, 32]. For example, under the proposed condition in this
study, fructose can be detected at pH 6.0, while fructose
and glucose can be detected at pH 9.0.
The analytical features and method validations including
linearity, precisions (intra-day and inter-day), and recovery
were investigated. Analytical curves for fructose and
mixture (1:1 of fructose and glucose) were obtained by
plotting the absorbance (at 350 nm) versus concentrations of
the sugars. The curves (Fig. 7) are linearizable, ranged from
10 to 50 mg L
-1
, with the correlation coefcient (r
2
) higher
than 0.99. Intra-day (n = 3) and inter-day (n = 3 9 3 days)
precisions were evaluated by replicate measurements of
the absorbance for fructose (15 mg L
-1
) and mixture
(15 mg L
-1
each), and expressed in terms of relative stan-
dard deviation (RSD). It was found that good precisions with
RSD lower than 1.5 and 5.0% for intra-day and inter-day
reproducibility were obtained.
In order to evaluate the accuracy of the method, the
recovery was studied by spiking standard glucose and
fructose (15 mg L
-1
each) into the samples [Beneo HP and
Jerusalem artichoke (CN 52867)] before extraction and
analysis by spectrophotometry. Good recoveries (on aver-
age) of both fructose and glucose were obtained in range of
96.899.8%.
Quantication of inulin in Jerusalem artichoke tubers
To simplify the quantication of inulin, the content of
inulin can be calculated based only on the fructose content,
as described by some researchers [20, 33]. The following
equation was used:
I kF
tot
F
f
4
where I is the inulin content, F
tot
is total fructose content,
F
f
is free fructose, and k is a correction factor for the
glucose part of the inulin and for the water loss during
hydrolysis. In this study, k = 0.995 is adopted, this value is
recommended for the unknown DP inulin [20]. The results
are summarized in Table 1, and it was found that inulin
content in artichoke tuber samples was in the range
62.9674.90% dry weight, along with free fructose con-
centration in the range of 1.181.60% dry weight. To
Fig. 6 Inuence of reaction time for a periodate and b iodide
reactions with sugars
Fig. 7 Analytical curves of fructose and mixture (1:1, fructose and
glucose)
614 Eur Food Res Technol (2011) 233:609616
1 3
establish the reliability of the proposed method for the
analysis of inulin in the artichoke samples, the results
obtained using spectrophotometry were compared to that
using HPAEC-PAD (Table 1), where both glucose and
fructose were quantied. It is clearly seen (from HPAEC-
PAD) that glucose is detected in trace amounts compared
to the majority component fructose, where glucose was
found below 5.0% dry weight. The paired t test at 95%
condence limit (p = 0.05) showed insignicant differ-
ences between the results obtained from spectrophotometry
and from HPAEC-PAD. In addition, the contents of inulin
found in the Jerusalem artichoke samples studied are in
good agreement with the results reported by others.
Fleming and Wassink [34] reported the content of fructans
(or inulin) in Jerusalem artichoke tuber in the range of
6883% dry weight.
Evaluation of degree of polymerization
To evaluate the degree of polymerization (DP) of inulin,
the results were obtained using HPAEC-PAD. The average
degree of polymerization (DP) was deduced using the
following equation [5, 33]:
DP number of Funits per Gunit 1 Gunit 5
where F and G are fructose and glucose, respectively. In this
study, the contents of fructose and glucose were obtained
from HPAEC-PAD, in which the calibration curves (peak
area versus concentrations of glucose and fructose stan-
dards) were used. A DP of 26.5 was obtained for the Beneo
HP inulin standard (calculated from the content of glucose
8.04 mg L
-1
and fructose 205.59 mg L
-1
), relatively close
to the labeled value (DP = 25). The average DP of inulin in
the samples was 1420, which is comparable to the DP
values reported in the published papers: 1620 [2], 7.511.2
[25], and 1020 [35]. However, the value of the DP is
depended on species, cultivar, production conditions,
physiological age, and other factors [4, 26].
Conclusions
This study describes the indirect determination method for
the quantication of inulin from Jerusalem artichoke
tubers. The tubers were extracted using pressurized liquid
extraction and the resulting inulin extracts were hydrolyzed
with acid. The amount of inulin-derived fructose in the
hydrolysates were then spectrophotometrically detected
using a periodate reaction. The content of inulin was then
deduced on the basis of the fructose content. The method
was applied for the determination of inulin in Jerusalem
artichoke tubers grown in Thailand. The inulin contents in
11 varieties of the selected artichokes were relatively
similar, with average inulin content of 71% dry weight.
The proposed method is rapid, simple, and reliable for
extraction and analysis of inulin in Jerusalem artichoke
samples. Moreover, it is relatively inexpensive compared
to the enzymatic hydrolysis method. The method is suitable
for routine analysis of Jerusalem artichoke, especially for
plant breeding purposes.
Acknowledgments Financial support from the Center for Innova-
tion in Chemistry (PERCH-CIC), Commission on Higher Education,
Ministry of Education is gratefully acknowledged. Grateful
acknowledgement for nancial support is also made to the Thailand
Research Fund (TRF), the Commission for Higher Education (CHE)
through the Distinguish Research Professor Grant of Professor
Dr.Aran Patanothai and the Higher Education Research Promotion
and National Research University Project of Thailand , Ofce of the
Table 1 Content and average DP of inulin from Jerusalem artichoke samples obtained from spectrophotometry and HPAEC-PAD
Jerusalem artichoke sample Content of sugar (dry wt. %)
Spectrophotometer Anion exchange chromatography
F
f
F I F
i
F G I Average DP
HEL 61 1.25 0.05 66.17 0.54 64.59 1.12 0.05 66.10 0.65 4.93 0.21 64.65 14.4
HEL 65 1.50 0.06 75.95 0.54 74.07 1.44 0.06 75.94 0.79 4.97 0.19 74.12 16.3
HEL 66 1.30 0.03 67.04 0.66 65.41 1.15 0.03 66.66 0.31 4.62 0.24 65.18 15.4
HEL 69 1.50 0.04 74.80 0.73 72.93 1.61 0.04 73.95 0.53 4.89 0.70 71.97 16.1
HEL 231 1.48 0.04 76.76 0.67 74.90 1.33 0.05 76.52 0.80 4.10 0.28 74.81 19.7
HEL 335 1.60 0.05 65.66 0.20 63.73 1.52 0.05 64.29 0.37 3.88 0.49 62.45 17.6
JA 37 1.40 0.06 74.67 0.67 72.90 1.29 0.07 74.03 0.61 5.03 0.41 72.37 15.7
JA 38 1.22 0.04 73.21 0.20 71.63 1.13 0.05 74.70 0.25 4.28 0.18 73.20 18.5
JA 102 1.18 0.03 71.44 0.27 69.90 1.10 0.05 72.63 0.11 4.58 0.12 71.17 16.9
KKU AC 001 1.23 0.02 64.51 0.48 62.96 1.17 0.02 64.09 0.69 3.58 0.45 62.60 18.9
F is total content of fructose, G is glucose, I is inulin, and DP is degree of polymerization of inulin
Eur Food Res Technol (2011) 233:609616 615
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Higher Education Commission, through the Food and Functional
Food Research Cluster of Khon Kaen University. The valuable sug-
gestions from Prof. Richard L. Deming (California State University of
Fullerton, USA) are also acknowledged.
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