Article

pubs.acs.org/JAFC

Determination of Inulin-type Fructooligosaccharides in Edible Plants

by High-Performance Liquid Chromatography with Charged Aerosol
Detector
Jing Li, Dejun Hu, Wanrong Zong, Guangping Lv, Jing Zhao,* and Shaoping Li*
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao
Special Administrative Region, China
ABSTRACT: Fructooligosaccharides (FOS), which are regarded as functional ingredients, are commonly classied as dietary
bers in many countries. However, few analytical methods for separation and analysis of individual FOS in plants, crops, and food
products have been developed. In this study, a simple, rapid, and sensitive high performance liquid chromatography with charged
aerosol detector (HPLC-CAD) method was developed for simultaneous determination of 11 inulin-type FOS with degree
of polymerization (DP) 313 in dierent samples. The separation was performed on a Waters XBridge Amide column
(4.6 250 mm i.d., 3.5 m) with gradient elution. All calibration curves for investigated analytes showed good linear regression
(R2 > 0.9962). Their limits of detection (LOD) and quantication (LOQ) were in the ranges 0.40.6 g/mL and 1.42.3 g/mL,
respectively. The recoveries ranged from 94.0% to 114.4%. A liquid chromatographytandem mass spectrometry (LC-MS/MS)
method was applied to qualitative analysis of FOS in dierent samples. The developed method was successfully applied to analysis
of 11 FOS in dierent samples of plants from Compositae, Campanulaceae, and Rubiaceae families. The developed HPLC-CAD
nethod with microwave-assisted extraction can be used for quantitative analysis of FOS and is helpful for quality control of plants
containing FOS.
KEYWORDS: HPLC-CAD, microwave-assisted extraction, quantitative analysis, inulin-type fructooligosaccharide

or evaporative light scattering detection (ELSD),2123 have

been developed for the determination of FOS. However,
colorization methods can determine only the content of total
sugars and the specicity is poor. HPAEC needs specic
instruments and columns, which are resistance to alkali. In
addition, mass spectrometry (MS) is incompatible with the
highly alkaline eluents used in HPAEC. Compared to HPAECPAD, HILIC can provide better results for analysis of mixtures,
including oligosaccharides of dierent degrees of polymerization.24 HPLC-RID method usually suers from poor
resolution because gradient elution is not available for RID.
The RID and ELSD also show poor sensitivity of detection,
especially for FOS with high degrees of polymerization (DP).
Until now only FOS (DP below10) have been determined
in plants, so quantitative analysis of FOS is still dicult, owing
to poor sensitivity and/or the absence of standards with
higher DP.
Recently a new type of universal detector, the so-called
charged aerosol detector (CAD), has been developed and
introduced by Dixon and Peterson.25 Similar to ELSD, CAD
provides universal detection for semivolatile or nonvolatile compounds and the response is independent of analyte structural
properties with or without a strong chromophore.26 However,
generally, CAD is more sensitive than ELSD, about 26 times
better than that of ELSD.26,27 Recently, it has been used as a
powerful tool for determination of oligosaccharides,28 lipids,29

INTRODUCTION
Fructooligosaccharides (FOS) are ocially recognized as
natural food ingredients and classied as dietary ber in almost
all European countries.1 The average daily consumption has
been estimated to be between 3 and 11 g in Europe2 and
between 1 and 4 g in the United States.3 FOS possess many
bioactive characteristics, such as prebiotic eects, suppressing
putrefactive pathogens, reducing the risk of colon cancer,
cognitive improvement and cerebral protective eects, and
decreasing the levels of blood glucose, serum cholesterol,
phospholipids, and triglycerides.1,49 In addition, FOS was
also a promising elicitor in postharvest disease control in various
fruits.10
Fructooligosaccharides are found in a number of mono- and
dicotyledonous families such as Liliaceae, Amaryllidacea,
Gramineae, and Compositae.11 They naturally exist in a large
variety of edible and medicinal plants used as functional foods,
such as Atractylodes macrocephala Koidz., Platycodon grandiorum (Jacq.) A.DC., Helianthus tuberosus L., Smallanthus
sonchifolius (Poepp. & Endl.) H. Robinson, Morinda ocinalis
How, and Arctium lappa L.1217 Actuarially, these medicine and
food dual-purpose plants, which are abundant in FOS, have
been cultivated and used as vegetable or medicated diet for
hundreds of years in China. Therefore, quantitative analysis of
FOS is very necessary and important for quality control of FOS
in these plants.
A series of methods, including colorization (phenolsulfuric
acid, dinitrosalicylic acid) methods,1 high-performance anion
exchange chromatography with pulsed amperometric detection
(HPAEC-PAD),1820 and hydrophilic interaction chromatography (HILIC) coupled with refractive index detection (RID)
2014 American Chemical Society

Received:
Revised:
Accepted:
Published:
7707

May 20, 2014

July 17, 2014
July 18, 2014
July 18, 2014
dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

saponins,26 and pharmaceutical drugs.30 Currently, there are no

reports on the application of CAD to the analysis of FOS.
Here, a HPLC-CAD and microwave-assisted extraction
method was developed for simultaneous determination of 11
FOS with DP 313 in 12 dierent plants in Compositae,
Campanulaceae, and Rubiaceae families. The methods were
validated and demonstrated for use in analysis of inulin-type
FOS and are potentially useful for other plants.

these samples were deposited at the Institute of Chinese Medical

Sciences, University of Macau, Macao SAR, China.
Inulin-type FOS with DP between 3 and 13 (all purities determined
by HPLC-DAD-ELSD were more than 95%) were separated and
puried in our laboratory (Figure 1). The structures were conrmed

MATERIALS AND METHODS

Materials, Reagents, and Chemicals. The analyzed plants

belonged to Compositae, Campanulaceae, and Rubiaceae families,
and their detailed characteristics are presented in Table 1. Species
identication was performed by Dr. C. F. Qiao. Voucher specimens of

Table 1. Characteristics of Analyzed Samples

producing area

place of purchase

codes

Atractylodes lancea (Thunb.) DC., family Compositae

Jiangsu
Shenzhen
A1
Hebei
Zhuhai
A2
unknown
Macau
A3
Codonopsis pilosula (Franch.) Nannf., family Campanulaceae
Gansu
Shenzhen
B1
Shanxi
Zhuhai
B2
unknown
Macau
B3
Atractylodes macrocephala Koidz., family Compositae
Anhui
Shenzhen
C1
Hunan
Zhuhai
C2
unknown
Macau
C3
Taraxacum mongolicum Hand.-Mazz., family Compositae
Hunan
Shenzhen
D1
Shanxi
Zhuhai
D2
Unkonwn
Macau
D3
Platycodon grandiorus (Jacq.) A.DC., family Campanulaceae
Anhui
Shenzhen
E1
unknown
Zhuhai
E2
unknown
Macau
E3
Aucklandia lappa Decne., family Compositae
Guangdong
Shenzhen
F1
Yunnan
Zhuhai
F2
unknown
Macau
F3
Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson, family Compositae
Guangdong
Zhuhai
G1
Guangdong
Macau
G2
Carthamus tinctorius L., family Compositae
Sichuan
Shenzhen
H1
Sichuan
Zhuhai
H2
unknown
Macau
H3
Adenophora tetraphylla (Thunb.) Fisch., family Campanulaceae
Anhui
Shenzhen
I1
unknown
Macau
I2
Morinda ocinalis How., family Rubiaceae
unknown
Zhuhai
J1
unknown
Meizhou
J2
Deqing
Deqing
J3
Helianthus tuberosus L., family Compositae
Hunan
Hunan
K1
Arctium lappa L., family Compositae
Korea
Macau
L1
Shandong
Shandong
L2
Jilin
Jilin
L3

Figure 1. Structures of inulin-type fructooligosaccharides (FOS).

by comparing their methylation, MS, and NMR data with the
literature.17,22 Acetonitrile for HPLC was purchased from Merck
(Darmstadt, Germany). Puried water for HPLC was prepared
on a Millipore Milli-Q Plus system (Millipore, Bedford, MA). All
other chemicals and reagents were of analytical grade.
Preparation of Standard Solutions. Mixed standard stock
solution containing inulin-type FOS (DP3DP13) was prepared
in 60% ethanol. The concentrations of DP3DP13 were about
20 mg/mL. The standard stock solution was stored in a refrigerator
at 4 C before use. Working standard solutions were prepared from
the stock solution by dilution with the appropriate volume of 60%
ethanol.
Sample Preparation. Extraction experiments were carried out
with a Multiwave 3000 reaction system (Anton Paar GmbH,
Graz, Austria) equipped with a 64-position extraction rotor. The
herbal medicine powder (0.1 g, 40 mesh) and 2 mL of ethanol
water (60:40 v/v) were transferred into 5 mL extraction vessels
made of borosilicate glass. After the vessels were closed and capped,
the rotor was placed inside the microwave oven. The extractions
were performed in temperature-controlled mode. After a heating ramp of 1 min, the microwave program was stated as 400 W for
5 min at the temperature of 80 C. Immediately after the temperature program had nished, all vessels were cooled down to
ambient temperature. All extractions were performed with magnetic stirring. After extraction, an aliquot was taken and centrifuged
for 5 min at 5000g. After centrifugation, 1 mL of supernatant
was transferred into a 10 mL volumetric ask and made up to the
volume with extraction solvent. Then the solution was ltered
through a 0.45 m lter before injection into the HPLC system for
analysis.
HPLC-CAD Analysis. Chromatographic analyses were performed
on a Dionex (Germering, Germany) Ultimate 3000 UHPLC system
equipped with Ultimate 3000 degasser, pump, RS autosampler, and RS
column compartment, coupled with a Corona charged aerosol detector
(CAD) instrument (ESA, Chelmsford, MA). Data processing was
carried out with Chromeleon 6.8 software (Dionex). The N2 pressure
of the CAD was adjusted to 35 psi and the response range was set
to 100 pA. Separations were carried out on a Waters XBridge Amide
column (4.6 250 mm id, 3.5 m). The column temperature was set
at 30 C. The mobile phase was consist of water (A) and acetonitrile
7708

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

Table 3. Precision (Intra- and Interday) for

Fructooligosaccharides DP3DP13
peak area (% RSD)

retention time (% RSD)

analyte

intraday
(n = 6)

interdaya
(n = 6)

intraday
(n = 6)

interdaya
(n = 6)

DP3
DP4
DP5
DP6
DP7
DP8
DP9
DP10
DP11
DP12
DP13

2.75
2.40
2.10
1.48
1.67
1.58
1.57
1.80
1.72
1.90
2.62

11.10
10.20
9.90
9.56
11.76
11.64
13.05
13.57
13.17
17.33
15.74

1.50
1.30
1.13
1.03
0.95
0.89
0.85
0.81
0.76
0.75
0.72

0.05
0.06
0.05
0.06
0.07
0.06
0.05
0.07
0.06
0.05
0.06

Interday precision was determined over a period of 5 days

(3 measurements on each day).

Figure 2. Eect of extraction time on extraction eciency of FOS with

dierent degrees of polymerization (DP).

Table 4. Repeatability, Stability, and Accuracy for

Fructooligosaccharides DP3DP13

Table 2. Linear Regression Data and Limits of Detection and

Quantitation for Fructooligosaccharides DP3DP13
analyte
DP3
DP4
DP5
DP6
DP7
DP8
DP9
DP10
DP11
DP12
DP13

R2

linear range
(g/mL)

LOD
(g/mL)

LOQ
(g/mL)

0.9993

1.453.8

0.4

2.3

0.9979

1.658.5

0.6

1.9

0.9978

1.452.5

0.6

1.4

0.9983

1.536.7

0.4

1.5

0.9968

1.538.3

0.5

1.5

0.9962

1.538.0

0.4

1.6

0.9967

1.537.5

0.4

1.6

0.9981

1.434.7

0.4

1.4

0.9971

1.4106.5

0.4

1.4

0.9978

1.396.0

0.6

1.6

0.9981

1.5221.0

0.5

1.5

regression eq
y = 1.3400x
2.3594
y = 1.3437x
2.2999
y = 1.3309x
2.2231
y = 1.3285x
2.1593
y = 1.3112x
2.232
y = 1.2913x
2.1935
y = 1.3084x
2.2270
y = 1.3165x
2.2200
y = 1.2250x
2.1673
y = 1.2024x
2.2063
y = 1.1955x
2.2064

repeatability

stability

accuracy

analyte

content (mg/g)

RSD (%)

RSD (%)

recovery (%)

RSD (%)

DP3
DP4
DP5
DP6
DP7
DP8
DP9
DP10
DP11
DP12
DP13

19.28
29.74
31.72
30.14
35.25
32.51
28.52
23.45
25.62
24.37
19.39

2.44
2.18
2.11
2.07
2.03
2.25
2.28
2.44
3.09
2.93
2.94

2.21
2.36
3.53
3.49
3.36
3.48
3.71
3.54
4.46
4.97
4.88

99.0
100.4
95.9
97.5
94.0
94.1
95.6
98.7
113.3
114.4
99.5

3.92
4.43
5.26
4.44
5.18
5.07
5.00
5.17
5.36
5.40
7.25

chromatographic conditions were determined at a signal-to-noise ratio

(S/N) of 3 and 10, respectively.
Precision, Repeatability, Accuracy, and Stability. Precision
of the HPLC-CAD method was determined by evaluating the
repeatability of intraday and interday measurements. Intraday
precision was determined by repeating the analysis of the mixed
standards solution for six replicates within 1 day, while interday
precision was determined in duplicates on three successive days.
Repeatability of the developed method was conrmed with
preparation and analysis of six parallel samples. Stability was tested
and analyzed at 0, 1, 2, 4, 8, 12, and 24 h. In order to check
the accuracy of the developed method, recovery experiments were
carried out as follows: accurate amounts of the standards were
spiked into the samples (0.05 g) in the form of solution. The spiked
samples were extracted, processed, and quantied in accordance
with methods mentioned above. Average recoveries were determined
by the equation recovery (percent) = 100 (observed amount
original amount)/spiked amount, and relative standard deviation
(RSD, percent) = 100 standard deviation (SD)/mean.
Liquid ChromatographyTandem Mass Spectrometric
Analyses. LC-MS/MS analyses were carried out on an Agilent 1100
Series LC/MSD Trap system (Agilent Technologies, Palo Alto, CA),
equipped with vacuum degasser, quaternary pump, autosampler,
column compartment, diode-array detector (DAD), and ion-trap
mass spectrometer with electrospray ionization (ESI) interface,
connected to Agilent LC/MSD Trap software. ESI-MS conditions
were as follows: drying gas N2 at 7 L/min, temperature 325 C,
pressure of nebulizer 25 psi, source voltage 3.5 kV. ESI-MS/MS

(B) with gradient elution: 75%45% B at 030 min, 45%75% B at

3032 min, and then equilibrated with 75% B for 10 min. The ow
rate was 1.0 mL/min and injection volume was 5 L.
Calibration Curves and Limits of Detection and Quantication. Standard stock solutions containing reference compounds were
prepared and diluted to appropriate concentrations for construction of calibration curves. Six concentrations of 11 analyte solutions
were injected in triplicate, and then the calibration curves were
constructed and their linear ranges determined. Since CAD response
was nonlinear and it can be represented by y = axb, where y refers
to peak area, while x is the analyte amount, a is the constant term,
and b is the exponential response factor.31 Therefore, calibration
of CAD was constructed by a double logarithmic plot as in previous
reports.26,29,31
Stock solutions were diluted to a series of appropriate concentration
with 60% ethanol, and aliquots of the diluted solutions were injected into HPLC for determining the limit of detection (LOD) and limit
of quantication (LOQ). The LOD and LOQ under the present
7709

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

Figure 3. (a) LCMS/MS spectra and (bd) Domon and Costello nomenclature for the fragmentation of separated FOS (DP3DP10).

conditions included isolation width 4 and fragment amplication

1.5. Scan ranges of both MS and MS/MS were 2002200 m/z. The
liquid chromatographic conditions for LC-MS/MS were the same as
described for HPLC-CAD analysis.

RESULTS AND DISCUSSION

Optimization of Sample Preparation. In order to obtain

quantitative extraction, the extraction method, extraction solvents,

7710

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

respectively (Table 2). For intraday precision, RSDs of the peak

area was 1.482.75% and RSD of the retention time was 0.72
1.50%; for interday precision, the corresponding RSDs were
9.5617.33% and 0.050.07% (Table 3). Recoveries ranged
from 94.0% to 114.4%, and the analytes were very stable in 60%
ethanol solution during the tested period (Table 4). The high
recoveries (113.3% and 114.4%) of DP11 and DP12 might be
attributed to the variation of their purities, and more attention
should be paid in future.
LC-MS/MS Analysis. Molecular mass information on the
standards and the extracted samples were determined in negative ion mode. In the negative ion mode, the [M H] of FOS
(DP3DP13) were detected at m/z 503, 665, 827, 989, 1151,
1313, 1475, 1637, 1799, 1961, and 2123. ESI-MS/MS of their
[M H] ions was performed to obtain the individual fragment patterns. Their MS/MS spectra are shown in Figure 3.
The fragment ions of FOS include glycosidic cleavages between
two sugar residues and cross-ring cleavages of two bonds within
the sugar ring. In our MS/MS spectra, the fragment ions of
FOS mainly result from glycosidic cleavages between two sugar
residues, which were as the same as those reported in previous
literature.17 That little fragments with low abundance of the
cross-ring cleavages of two bonds within the sugar ring were
found might be derived from the low collision energy or few
sodium ions in mobile phase.17,32 FOS (DP11DP13) with
high molecular weights are dicult to ionize for LC-MS/MS
analysis, so the characteristic MS/MS spectra of FOS (DP11
DP13) could not be shown. However, in the MS spectrum of
FOS (DP11DP13), the characteristic ions at m/z 161, 179,
323, 341, 485, 503, 647, 665, 809, 827, 971, 989, 1133, 1151,
1313, 1475, 1457, 1638, and 1800 were obtained. In summary,
all the results suggested that FOS should consist of the corresponding number of hexoses.
Quantitative Determination of FOS in Plants. The
established HPLC-CAD method was applied for analysis of
inulin-type FOS (DP3DP13) in dierent samples of Compositae, Campanulaceae, and Rubiaceae families. HPLC-CAD
chromatograms of the mixed standards and dierent samples
are shown in Figure 4. Besides the investigated 11 oligosaccharides, many oligosaccharides with DP of more than 13
were also found in these samples. Unfortunately, we could not
quantitatively determine these oligosaccharides because of
the absence of related reference compounds, which should be
performed in the future. The contents of the 11 investigated FOS in dierent samples are summarized in Table 5 and
Figure 5. The results showed that the three samples with
highest contents of FOS were Morinda ocinalis, Helianthus
tuberosus and Arctium lappa, and both H. tuberosus and A. lappa
belong to the Compositae family. The data also indicated there
was no FOS detected in Taraxacum mongolicum and Carthamus
tinctorius, which are also of the Compositae family. That is
because FOS are usually the chemical compounds for storing
energy in organs such as bulbs, tubers, and tuberous roots. The
owers contain little FOS.
In summary, there are many reports on the existence of FOS
in plants of Liliaceae, Compositae, Campanulaceae, and
Rubiaceae families, but few reports on their contents in these
samples. This is the rst report for quantitative analysis of
inulin-type FOS (DP3DP13) in dierent samples of plants in
Compositae, Campanulaceae, and Rubiaceae families, which is
helpful to improve their qualities. The developed HPLC-CAD
method, which is accurate and specic, can also be used for
determination of inulin-type FOS in other plants.

Figure 4. HPLC-CAD chromatograms of (A) mixed standards and

(BD) samples of (B) Atractylodes macrocephala, (C) Platycodon
grandiorum, and (D) Morinda ocinalis.

extraction solvent volume, extraction time, and temperature were

optimized. Solvent choices, solvent volumes, and temperature
used in this study were xed according to our previous study.23
We investigated the eect of extraction time, which directly
inuence the extraction yields of FOS.
In order to get the optimal reaction time, we performed the
extraction by microwave extraction in a range from 2 to 20 min
at 80 C, using a sample (Morinda ocinalis, sample J3) of
approximately 0.1 g. Results (Figure 2) showed that the optimal
extraction time of FOS was 5 min, whereas longer extraction
time did not signicantly increase extraction yields and FOS
degradation was observed at 20 min. We therefore selected 5
min as the optimal extraction time.
Optimization of HPLC Conditions. Analysis of FOS is a
challenge due to their high polarity and complexity and lack of
specic UV absorptivity. Therefore, the HPLC-CAD method
was selected for analysis of FOS. Several dierent commercial
columns were tested, and nally the Waters XBridge Amide
column (4.6 250 mm id, 3.5 m) was selected for separation of 11 investigated inulin-type FOS because of its high
resolution and good signal-to-noise performance. The analytes
in both mixed standard solution and sample solutions can be
well separated by gradient elution with water and acetonitrile.
Method Validation. Linearity, regression, and linear ranges
of the 11 analytes were determined by the developed HPLC
method. The data indicated the calibration curves of the 11
analytes had good linearity (R2 > 0.9962), and their LODs and
LOQs were in the ranges 0.40.6 g/mL and 1.42.3 g/mL,
7711

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

Table 5. Contents of the 11 Analytes in Tested Samples

analyte content(mg/g)

sample

DP3

DP4

DP5

DP6

DP7

DP8

DP9

DP10

DP11

DP12

DP13

total

A1
A2
A3
B1
B2
B3
C1
C2
C3
D1a
D2a
D3a
E1
E2
E3
F1
F2
F3
G1
G2
H1a
H2a
H3a
I1
I2
J1
J2
J3
K1
L1
L2
L3

6.01
4.68
2.69
1.49
3.59
0.65
4.29
4.52
7.15

4.05
2.86
2.14
0.8
1.62
0.42
4.08
4.36
5.65

3.27
1.96
1.66
0.75
1.25
0.39
4.39
4.78
5.4

4.22
3.24
1.7
1.34
3.01
0.4
4.42
4.91
5.22

5.08
3.85
2.09
1.48
3.15
0.48
5.61
6.28
6.68

5.44
2.96
2.22
1.48
2.81
0.51
5.77
6.57
6.7

5.59
3.08
2.5
1.59
2.49
0.64
6.03
6.89
6.92

5.59
3.02
2.75
1.64
1.99
0.76
6.06
6.69
6.7

6.63
3.48
3.63
2.13
2.19
1.05
7.32
7.79
7.83

7.56
3.88
4.33
2.46
2.2
1.34
8.41
8.76
8.72

7.46
3.3
4.64
2.35
1.91
1.38
8.24
8.3
8.41

60.89
36.32
30.36
17.52
26.21
8.01
64.62
69.84
75.38

4.02
4.8
8.49
4.5
4.28
2.92
29.43
23.8

4.71
4.67
6.58
2.97
2.59
2.25
27.17
24.54

4.93
4.28
6.56
1.96
1.85
1.65
19.8
18.91

5.67
4.52
7.14
2.23
1.78
2.13
14.17
13.92

7.47
5.55
9.42
2.28
1.89
2.5
13.04
15.25

8.01
5.61
10.07
2.33
1.83
2.46
11.09
14.32

8.78
5.48
10.74
2.74
2.05
2.74
9.36
13

8.97
5.93
10.75
2.85
2.36
3.2
7.1
10.35

11.19
7.47
13.52
3.47
3.14
4.21
6.06
9.61

12.93
9.22
15.65
4.22
4.28
5.17
4.75
7.89

12.65
9.38
15.6
4.32
4.51
5.49
3.17
5.57

89.34
66.91
114.52
33.86
30.55
34.72
145.13
157.17

4.28
5.17
22.28
11.92
19.58
36.42
24.05
22.8
16.52

4.69
4.98
28.97
18.02
29.24
28.58
22.08
18.59
13.33

4.22
5.65
30.34
20.53
31.61
23.82
15
16.06
12.87

4.47
5.97
29.16
20.08
29.89
19.6
15.7
14.86
11.71

4.81
8.11
32.11
23.93
35.06
19.85
18.54
17.21
13.97

4.21
9.01
27.47
22.5
32.44
18.57
17.68
14.57
13.72

4.13
10.11
22.93
20.3
28.46
16.16
15.81
13.52
13.11

4.11
10.62
18.36
17.39
23.61
15.13
13.54
11.72
11.87

5.22
13.92
19.35
19.6
25.56
15.54
14.76
12.06
13.93

6.59
16.7
18.62
19.88
24.47
14.54
14.74
12.82
15.44

7.31
17.08
15.52
17.11
19.54
11.81
13.22
11.37
14.78

54.04
107.31
265.11
211.24
299.44
220.02
185.13
165.59
151.26

Not detected.

* Phone +853-8397-4692; fax +853-2884-1358; e-mail spli@

umac.mo.
Author Contributions

J.L., D.H., and W.Z. contributed equally to this work

Funding

This research was supported by grants from the Science and

Technology Development Fund of Macao (FDCT059/2011/A3),
and the University of Macau (MYRG140 and MYRG085).
Notes

The authors declare no competing nancial interest.

ABBREVIATIONS
HPLC, high-performance liquid chromatography; CAD,
charged aerosol detector; FOS, fructooligosaccharides; DP,
degree of polymerization; ELSD, evaporative light scattering
detector; HPAEC, high-performance anion-exchange chromatography; PAD, pulsed amperometric detection; RID, refractive
index detector; MS, mass spectrometry; LOD, limit of
detection; LOQ, limit of quantication; S/N, signal-to-noise
ratio

Figure 5. Average contents of FOS with dierent degree of polymerization (DP) in dierent edible plants. AL mean the average of
three related samples in Table 1.

AUTHOR INFORMATION

Corresponding Authors

REFERENCES

(1) Milani, E.; Koocheki, A.; Golimovahhed, Q. A. Extraction of

inulin from Burdock root (Arctium lappa) using high intensity
ultrasound. Int. J. Food Sci. Technol. 2011, 46, 16991704.

* Phone +853-8397-4692; fax +853-2884-1358; e-mail

zhaojing.cpu@163.com.
7712

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713

Journal of Agricultural and Food Chemistry

Article

(2) Van Loo, J.; Coussement, P.; De Leenheer, L.; Hoebregs, H.;
Smits, G. On the presence of inulin and oligofructose as natural
ingredients in the Western diet. Crit. Rev. Food Sci. 1995, 35, 525552.
(3) Moshfegh, A. J.; Friday, J. E.; Goldman, J. P.; Chug Ahuja, J. K.
Presence of inulin and oligofructose in the diets of Americans. J. Nutr.
1999, 129, 1407S1411S.
(4) Pool-Zobel, B. L. Inulin-type fructans and reduction in colon
cancer risk: review of experimental and human data. Br. J. Nutr. 2005,
93, S73S90.
(5) Sangeetha, P. T.; Ramesh, M. N.; Prapulla, S. G. Recent trends in
the microbial production, analysis and application of fructooligosaccharides. Trends Food Sci. Technol. 2005, 16, 442457.
(6) Lou, Z.; Wang, H.; Wang, D.; Zhang, Y. Preparation of inulin and
phenols-rich dietary fibre powder from burdock root. Carbohydr.
Polym. 2009, 78, 666671.
(7) Zhang, R.; Zhou, J.; Jia, Z.; Zhang, Y.; Gu, G. Hypoglycemic
effect of Rehmannia glutinosa oligosaccharide in hyperglycemic and
alloxan-induced diabetic rats and its mechanism. J. Ethnopharmacol.
2004, 90, 3943.
(8) Li, Y.-F.; Li, Y.; Xu, Y.-K.; Yang, M.; Zhao, Y.-M.; Luo, Z.-P.
Antistress effect of oligosaccharides extracted from Morinda of ficinalis
in mice and rats. Acta Pharmacol. Sin. 2001, 22, 10841088.
(9) Li, Y.-F.; Liu, Y.-Q.; Yang, M.; Wang, H.-L.; Huang, W.-C.; Zhao,
Y.-M.; Luo, Z.-P. The cytoprotective effect of inulin-type hexasaccharide extracted from Morinda of f icinalis on PC12 cells against the
lesion induced by corticosterone. Life Sci. 2004, 75, 15311538.
(10) Sun, F.; Zhang, P.; Guo, M.; Yu, W.; Chen, K. Burdock
fructooligosaccharide induces fungal resistance in postharvest Kyoho
grapes by activating the salicylic acid-dependent pathway and
inhibiting browning. Food Chem. 2013, 138, 539546.
(11) Kaur, N.; Gupta, A. K. Applications of inulin and oligofructose
in health and nutrition. J. Biosci. 2002, 27, 703714.
(12) Flamm, G.; Glinsmann, W.; Kritchevsky, D.; Prosky, L.;
Roberfroid, M. Inulin and oligofructose as dietary fiber: A review of
the evidence. Crit. Rev. Food Sci. 2001, 41, 353362.
(13) Roberfroid, M. B. Functional foods: Concepts and application to
inulin and oligofructose. Br. J. Nutr. 2002, 87, S139S143.
(14) Zhu, H.-T.; Chen, J.-Y.; Chen, L.; Tu, Z.-L.; An, Z.-B.
Pharmacognosy studies on Atractylodes macrocephala Koidz. Lishizhen
Med. Mater. Med. Res. 2006, 17, 10191020.
(15) Fu, W.-W.; Dou, D.-Q.; Pei, Y.-H. Review on chemical
components and bioactivities of Platycodon grandif lorum. J. Shenyang
Pharm. Univ. 2006, 23, 184191.
(16) Yang, H.; Xie, J.-L.; Sun, H.-D. Research progress on the
medicinal plant Saussurea lappa. Nat. Prod. Res. Dev. 1997, 10, 9098.
(17) Yang, Z.; Yi, Y.; Gao, C.; Hou, D.; Hu, J.; Zhao, M. Isolation of
inulin-type oligosaccharides from Chinese traditional medicine:
Morinda of f icinalis How. and their characterization using ESI-MS/
MS. J. Sep. Sci. 2010, 33, 120125.
(18) Ishiguro, Y.; Ueno, K.; Onodera, S.; Benkeblia, N.; Shiomi, N.
Effect of temperatures on inulobiose and inulooligosaccharides in
burdock roots during storage. J. Food Compos. Anal. 2011, 24, 398
401.
(19) Ishiguro, Y.; Onodera, S.; Yoshihira, T.; Kosaka, S.; Benkeblia,
N.; Shiomi, N. Variation of 1-kestose, nystose and higher
fructooligosaccharides in burdock root stored under different temperature regimes. Agric. Biol. J. N. Am. 2010, 1, 491494.
(20) Ishiguro, Y.; Onodera, S.; Benkeblia, N.; Shiomi, N. Variation of
total FOS, total IOS, inulin and their related-metabolizing enzymes in
burdock roots (Arctium lappa L.) stored under different temperatures.
Postharvest Biol. Technol. 2010, 56, 232238.
(21) Imahori, Y.; Kitamura, N.; Kobayashi, S.; Takihara, T.; Ose, K.;
Ueda, Y. Changes in fructooligosaccaride composition and related
enzyme activities of burdock root during low-temperature storage.
Postharvest Biol. Technol. 2010, 55, 1520.
(22) Yang, Z.; Hu, J.; Zhao, M. Isolation and quantitative
determination of inulin-type oligosaccharides in roots of Morinda
of f icinalis. Carbohydr. Polym. 2011, 83, 19972004.

(23) Li, J.; Liu, X.; Zhou, B.; Zhao, J.; Li, S. Determination of
fructooligosaccharides in burdock using HPLC and microwave-assisted
extraction. J. Agric. Food Chem. 2013, 61, 58885892.
(24) Brokl, M.; Hernandez-Hernandez, O.; Soria, A. C.; Sanz, M. L.
Evaluation of different operation modes of high performance liquid
chromatography for the analysis of complex mixtures of neutral
oligosaccharides. J. Chromatogr. A 2011, 1218, 76977703.
(25) Dixon, R. W.; Peterson, D. S. Development and testing of a
detection method for liquid chromatography based on aerosol
charging. Anal. Chem. 2002, 74, 29302937.
(26) Eom, H. Y.; Park, S.-Y.; Kim, M. K.; Suh, J. H.; Yeom, H.; Min,
J. W.; Kim, U.; Lee, J.; Youm, J.-R.; Han, S. B. Comparison between
evaporative light scattering detection and charged aerosol detection for
the analysis of saikosaponins. J. Chromatogr. A 2010, 1217, 4347
4354.
(27) Vervoort, N.; Daemen, D.; Torok, G. Performance evaluation of
evaporative light scattering detection and charged aerosol detection in
reversed phase liquid chromatography. J. Chromatogr. A 2008, 1189,
92100.
(28) Inagaki, S.; Min, J. Z.; Toyooka, T. Direct detection method of
oligosaccharides by high-performance liquid chromatography with
charged aerosol detection. Biomed. Chromatogr. 2007, 21, 338342.
(29) Khoomrung, S.; Chumnanpuen, P.; Jansa-Ard, S.; Stashlman,
M.; Nookaew, I.; Boren, J.; Nielsen, J. Rapid quantification of yeast
lipid using microwave-assisted total lipid extraction and HPLC-CAD.
Anal. Chem. 2013, 85, 49124919.
(30) Pistorino, M.; Pfeifer, B. A. Polyketide analysis using mass
spectrometry, evaporative light scattering, and charged aerosol
detector systems. Anal Bioanal. Chem. 2008, 390, 11891193.
(31) Bai, C.-C.; Han, S.-Y.; Chai, X.-Y.; Jiang, Y.; Li, P.; Tu, P.-F.
Sensitive determination of saponins in Radix et Rhizoma notoginseng
by charged aerosol detector coupled with HPLC. J. Liq. Chromatogr.
Relat. Technol. 2008, 32, 242260.
(32) Reis, A.; Coimbra, M. A.; Domingues, P.; Ferrer-Correia, A. J.;
Rosario, M.; Domingues, M. Structural characterisation of underivatised olive pulp xylo-oligosaccharides by mass spectrometry using
matrix-assisted laser desorption/ionisation and electrospray ionisation.
Rapid Commun. Mass Spectrom. 2002, 16, 21242132.

7713

dx.doi.org/10.1021/jf502329n | J. Agric. Food Chem. 2014, 62, 77077713