Eur Food Res Technol
DOI 10.1007/s00217-016-2810-1
ORIGINAL PAPER
Isolation and identification of terpenoids from chicory roots
and their inhibitory activities against yeast α‑glucosidase
Hang Fan1 · Jian Chen1 · Han Lv1 · Xiancan Ao1 · Yuexian Wu1 · Bingru Ren1 ·
Weilin Li1 
Received: 5 July 2016 / Revised: 19 September 2016 / Accepted: 22 October 2016
© Springer-Verlag Berlin Heidelberg 2016
Abstract Chicory (Cichorium intybus L.var. sativum L.,
Asteraceae) is a multipurpose plant cultivated as vegeta-
ble and coffee substitute in Europe and North America,
also as folk medicine in China. The extracts from chic-
ory roots showed significant effect on inhibition against
α-glucosidase. By a bioassay-guided approach, the chemi-
cal fraction with high α-glucosidase inhibition was found
and its chemical profile was tentatively described by
UPLC-Q-TOF/MS to include 28 compounds. Further
chemical isolation yielded six compounds, their chemical
structures were elucidated as 11β-13-dihydrolactucin (1),
lactucin (2), 8-deoxylactucin (3), jacquinelin (4), 11β,13-
dihydrolactucopicrin (5) and lactucopicrin (6), and among
them, the compound 4 showed the strongest inhibitory
activity against yeast α-glucosidase with IC50 value of
4.180  μM. The present results suggest that chicory roots
can eventually exhibit anti-diabetic effect and the sesquiter-
penes may be responsible for the activity.
Keywords  Chicory (Cichorium intybus L. var. sativum
L.) · α-Glucosidase inhibition · Sesquiterpene
Introduction
Chicory (Cichorium intybus L. var. sativum L.) is a per-
ennial herb of the family Asteraceae cultivated mainly in
* Bingru Ren
bingruren@126.com
* Weilin Li
lwlcnbg@mail.cnbg.net
1
Institute of Botany, Jiangsu Province and Chinese Academy
of Sciences, Nanjing 210014, People’s Republic of China
Europe and North America for multipurpose use. Histori-
cally, chicory was grown as vegetable crop in Greek and
Roman nearly 4000 years ago [1], also as medicinal plant,
vegetable crop, coffee substitute and animal forage in
ancient Egyptian [2]. At present, chicory is widely culti-
vated in many places of the world, mainly Europe, India,
South Africa and North America for a wide use. The leaves
of chicory are used as salads, vegetable dishes and forage,
and its roots are used as a coffee substitute and additive and
for ethanol production by direct fermentation [3]. Chicory
pulp roots are also an important material in the inulin pro-
cessing industry. The extract of chicory roots are rich in
pectin, a polysaccharide extensively used in food as a gel-
ling agent, thickening agent and stabilizer [4].
Chicory is used as Uyghur folk medicine in China [5].
In recent years, some chemical components from chicory,
such as polysaccharides, triterpenes, flavonoids, phenolic
acids, were reported, and some of them showed various
bioactivities through in vivo and in vitro pharmacology
experiments [6], such as hepatoprotection [7], anti-bacterial
[8] and anti-hyperuricemia [9]. Up to now there was only
limited information about the anti-diabetic activity of chic-
ory, but not to mention its hypoglycemic constituents.
The α-glycosidase is the key enzyme responsible for
degradation of carbohydrates and production of glucose in
the small intestine [10]. Thus, α-glycosidase inhibitors are
clinically used to treat type 2 diabetes [11, 12]. This study
was aimed to estimate the inhibitory activity of chicory
against α-glycosidase and to reveal the effective compo-
nents for anti-diabetics. A bioassay-guided scheme was car-
ried out to estimate α-glycosidase inhibition of the different
fractions and subfractions. Afterward, the subfraction with
high α-glycosidase inhibition was characterized by using
UPLC-Q-TOF/MS for its chemical components, further
isolated and purified by using column chromatography of
13

silica gel, Sephadex LH-20 and prep-LC. The structures of
obtained compounds were then elucidated by nuclear mag-
netic resonance (NMR).
Materials and methods
Plant materials, chemicals and reagents
Fresh roots of chicory for chemical constituent investi-
gation were manually collected in July 2015 in Nanjing
Botanical Garden Mem. Sun Yat-sen, Nanjing, China. The
plant was identified by one of the authors (Professor Bing-
ru Ren). The roots were washed with running tap water,
dried at 45 °C and cut in small pieces. The obtained pieces
were then stored in a sealed polyethylene bag.
All solvents used for HPLC analysis were HPLC grade,
and others were analytical grade. HPLC-grade acetoni-
trile, methanol and formic acid used for HPLC analy-
sis and sample preparation were purchased from TEDIA
Scientific Products ((Fairfield, USA). Deuterated metha-
nol (CD3OD) and chloroform (CDCl3), the solvent for
NMR experiments, were bought from Shanghai Adamas
Reagent Co. Ltd. (Shanghai, China). p-Nitrophenyl-β-d-
glucopyranoside (pNPG), α-glucosidase (Type I) were pur-
chased from Sigma-Aldrich (Missouri, USA).
Extraction and isolation
To prevent daylight-induced lactone double-bond hydration
[13], dried roots of chicory were immersed in 80% EtOH,
with the soaking container light-protected and stored refrig-
erated for 3 weeks. The 80% EtOH extract was decanted,
filtered and evaporated under reduced pressure. Then, the
combined residues were successively partitioned with
petroleum ether (2 L, 3 times), ethyl acetate (2 L, 3 times)
and MeOH (2 L, 3 times) to obtain three fractions. Half of
the ethyl acetate fraction (EAFE, 21 g) was adsorbed on
100 g of silica gel (0.015–0.040 mm, Merck, Germany) and
further fractionated by column chromatography over silica
gel, eluted with a gradient solvent system with 10–100%
n-hexane/EtOAc (v/v) at an increment of 20% (2 L for each
gradient, 1.0 mL/min) under room temperature, to afford
six subfractions (Fr. A–Fr. F) based on HPLC spectra. Fr.
C was subjected to column chromatography over Sephadex
LH-20 gel (GE healthcare, Sweden) eluted with 20–100%
methanol/H2O (v/v) system at an increment of 20% (2 L
for each gradient, 17.9 mL/min) under room temperature
for further separation. According to the HPLC spectra of
elutes, subfractions Fr. C1–Fr. C5 were collected. Fraction
C1 was additionally purified by prep-HPLC on a Shimadzu
model LC-6AD pump equipped with a SPD-20A detec-
tor (Shimadzu Corporation, Japan) and a RP-C18 column
13
Eur Food Res Technol
(5  μm, 9.4 × 250 mm, Agilent, Germany) with an iso-
cratic elution of 40% MeOH/H2O (v/v) at a flow of 3.0 mL/
min to afford compound 1 (39.0 mg, tR  = 6.301 min)
and compound 2 (63.2 mg, tR  = 7.769 min). Fraction
C2 was additionally purified by prep-HPLC with 50%
MeOH/H2O (v/v) at a flow of 3.0 mL/min to obtain com-
pound 3 (36.0 mg, tR  = 11.838 min) and compound
4 (39.3 mg, tR  = 14.772 min). Fraction C4 was fur-
ther purified by prep-HPLC eluted with 60% MeOH/
H2O (v/v) at a flow of 3.0 mL/min to yield compound 5
(16.5 mg, tR  = 16.340 min) and compound 6 (7.8 mg,
tR  = 18.274 min). All the elution peaks were detected at
210 nm. Major peaks were obtained by UV absorbance
measurements at 210 nm. The flowchart is given in Fig. 1.
NMR studies
The nuclear magnetic resonance data were acquired on a
Bruker Advance 400 MHz NMR Spectrometer (Bruker
BioSpin GmbH, Germany) using tetramethylsilane (TMS)
as a reference at room temperature. 1H and 13C NMR spec-
tra were operated frequency of 400.13 and 100.57 MHz,
respectively. Samples were examined in CD3OD or CDCl3
solution in 5 mm i.d. tube (internal acetone 1H (CH3) at
2.1 ppm and 13C(CH3) at 31.5 ppm relative to TMS).
Activity estimation
The inhibitory activity against yeast α-glucosidase was
measured with a slight modification of the reported method
[14]. To each of a 96-well microtiter plate was added 4 μL of
α-glucosidase [1 U/mL in phosphate buffer (PBS), pH 6.8], 1
μL of diluted samples in dimethyl sulfoxide, 75 μL of PBS.
The mixture was incubated at 37 °C for 10 min, then 20 μL
p-nitrophenyl-β-d-glucopyranoside (pNPG) solution (3 mM
in PBS, pH 6.8) was added to each well, and the absorbance
at 405 nm was recorded with a microplate reader after incu-
bation at 37 °C for 30 min. The inhibition activity percentage
was calculated according to the following formula: inhibition
(%) = [1 − (As/Ac)] × 100%. As is the absorbance of the
sample, Ac is the absorbance of the control, and Acarbose
(Bayer, Germany) was used as a positive control.
ESI‑Q‑TOF–MS detection
EAFE was dissolved in methanol at a concentration of
5 g/L and filtered through a 0.45-μm PVD filter. The
mobile phases consisted of distilled water (A) and ace-
tonitrile (B). The following multistep linear gradient was
applied: 0 min, 10% B; 15 min, 30% B; 20 min, 40% B;
35 min, 60% B; 40 min, 70% B; 60 min, 100% B. The flow
rate was 1 mL/min, and the column temperature was at
35 °C. The injection volume was 10 μL.
Eur Food Res Technol
Fig. 1  Flowchart for the
bioassay-guided isolation of
active components against
α-glucosidase from chicory
roots
The UPLC-Q-TOF/MS analysis was carried out using
an Agilent 6530 accurate-mass quadrupole time-of-flight
system (Agilent, USA) under positive and negative ion
mode coupled with an Agilent ZORBAX SB-C18 column
(1.8 µm, 4.6 × 100 mm, Waldbronn, Germany), using a
capillary voltage of +4.0 kV. Nitrogen was used as nebuli-
zation and desolvation gas, and argon was used as the colli-
sion gas. The desolvation gas flow rate was set to 10 L/min
at temperature of 350 °C. The full-scan MS spectra were
acquired over a mass range of 100–1000 m/z. Analyses
were made using a Masshunter Qualitative Analysis soft-
ware (B.05.00).
Statistical analysis
All experiments were performed in triplicated (n = 3), and
all the data were expressed as mean ± square error (SD).
The IC50 values were defined as the concentration of sam-
ple inhibiting 50% of α-glucosidase, and calculated using
nonlinear regression algorithm (Origin 9.0, OriginLab Co.,
USA).
Results and discussion
Bioassay‑guided abstraction and isolation of active
components
As a result of preliminary screening of crude extracts
successively extracted by different solvents at same
100
80
Inhibition rate (%)
60
40
20
0
PEE EAFE ME Acarbose
Compounds
Fig. 2  Inhibitory activity of PEE, EAFE and ME extracted from
chicory roots against α-glucosidase
concentrations, petroleum ether extracts (PEE) and
methanol extracts (ME) were excluded, and ethyl acetate
extracts (EAFE) were subjected to ongoing study based
on their bioactivity (Fig. 2). EAFE was further isolated
by column chromatography on silica gel to obtain six
subfractions (Fr. A–Fr. F), which were evaluated for
α-glucosidase inhibitory activity. Figure 3 shows that
Fr. C yielded the highest α-glucosidase inhibition. That
was to say the compounds presented in Fr. C had better
α-glucosidase inhibition than that in other subfractions.
Therefore, Fr. C was chromatographed on a Sephadex
LH-20 column to obtain subfractions Fr. C1–Fr. C5. They
were further subjected to prep-HPLC to give compounds
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 Eur Food Res Technol

100 UPLC‑Q‑TOF/MS detection of EAFE

80 In order to understand approximately the bioactive com-

Inhibition rate (%) ponents in chicory, the detection of fraction with inhibi-
60 tory activity against α-glucosidase was conducted by using
UPLC-Q-TOF/MS. The total ion current chromatogram
40 of the fraction EAFE in positive mode is shown in Fig. 5,
and the compounds characterization is presented in Table 1
20 according to elution order. Twenty-eight compounds were
identified or tentatively characterized. Taking an example
0 for peak 6, the precise molecular weight was 425.1704,
Fr. A Fr. B Fr. C Fr. D Fr. E Fr. F Acar. and the main fragment ions that were analyzed via the MS/
Compounds
MS screening were observed at m/z 425.1704 [M + H]+,
263.1216 [M + H-Glc]+, 245.1206 [M + H-Glc-H2O]+ in
Fig. 3  Inhibitory activity of Fr. A–Fr. F extracted from chicory roots
against α-glucosidase
the positive-ion spectrum. Based on the analysis of elemen-
tal composition, fractional isotope abundance, the molecu-
lar formula of peak 6 was speculated to be C21H28O9, and
100 thus, the compound was inferred as crepidiaside B [15].
Among the compounds, fourteen compounds were dis-
80 covered as sesquiterpenes, including three sesquiterpene
glycosides, so it is considered that the sesquiterpenes
Inhibition rates(%)

60 may be the main ingredients for the inhibitory activity of

chicory root against α-glucosidase. This identification and
40 structural elucidation of the chemical constituents pro-
vides essential information for further isolation of active
20 components.

0 Identification of compounds
Acarbose1 2 3 4 5 6
Compounds The structure of six compounds isolated from active sub-
fraction Fr. C was elucidated by the comparison of their 1D
Fig. 4  Inhibitory activity of compounds isolated from chicory roots NMR and ESI–MS data with those in the literature.
against α-glucosidase
Compound 1 was isolated as a white powder, and its
ESI–MS analysis displayed a pseudomolecular ion peak
1, 2, 3, 4, 5 and 6. Inhibitory activities of compounds iso- [M  + H]+ at m/z 279.1, indicating that the molecular
lated from chicory against α-glucosidase are shown in weight of 1 was 278. In its 13C NMR spectrum, two car-
Fig. 4. bonyl carbons at δ 195.94 (C-2) and 178.59 (C-12), four

Fig. 5  Total ion chromatogram (TIC) of EAFE from chicory roots in positive mode

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Eur Food Res Technol

Table 1  Compounds identified in EAFE extracted from chicory roots

Peak No. T (min) Identification MS (error, ppm) Formula MS/MS (m/z) References

1 3.148 Not identified 851.2409 [M + H]+ C21H32O5 689.1934 [M + H–Glc]+

(4.30) 527.1450 [M + H–2Glc]+
365.0965 [M + H–3Glc]+
2 11.472 Chlorogenic acid 377.0739 [M + Na]+ C16H18O9 193.0555 [M + H–caffeoyl]+ [16]
355.094 [M + H]+ 175.0468 [M + H–caffeoyl–
H2O]+
353.0962 [M − H]−
(3.3)
3 13.736 11β,13-Dihydrolactucin (1) 301.0974 [M + Na]+ C15H18O5 261.105 [M + H–H2O]+ [17]
279.1167 [M + H]+ 215.1015 [M + H–H2O–
HCOOH]+
277.1144 [M − H]−
(2.58)
4 14.880 Lactucin (2) 299.0815 [M + Na]+ C15H16O5 259.0897 [M + H–H2O]+ [17]
277.1004 [M + H]+ 213.0859 [M + H–H2O–
HCOOH]+
275.0992 [M − H]−
(−1.03)
5 18.618 Not identified 701.4771 [M + Na]+ C33H48O18 515.1318 [M − H–Glc]−
679.4948 [M + H]+ 369.1410 [M–H–Glc–Rha]−
677.5186 [M − H]− 207.0693 [M–H–Glc–Rha–Glc]−
(0.21)
6 19.137 Crepidiaside B 447.1507 [M + Na]+ C21H28O9 263.1216 [M + H–Glc]+ [15]
425.1704 [M + H]+ 245.121 [M + H–Glc–H2O]+
423.1758 [M − H]−
(3.1)
7 19.606 Not identified 517.1215 [M + H]+ C21H24O15 499.2375 [M + H–H2O]+
515.1343 [M − H]− 337.186 [M + H–H2O–Glc]+
(−6.98)
8 20.183 Not identified 499.1117 [M + H]+ C21H22O14 337.1863 [M + H–Glc]+
497.2088 [M − H]− 319.172 [M + H–H2O–Glc]+
(4.77)
9 21.475 Cichorioside C 451.1253 [M + Na]+ C21H32O9 277.1920 [M + H–Glc]+ [18]
429.1437 [M + H]+ 259.176 [M + H–H2O–Glc]+
427.1496 [M − H]−
(3.27)
10 22.109 11β,13-Dihydrocichopumilide 251.1584 [M + H]+ C15H22O3 233.2304 [M + H–H2O]+ [19]
248.9773 [M − H]− 205.2290 [M + H–2H2O]+
(−4.1)
11 22.793 8-Deoxylactucin (3) 331.1462 [M + Na]+ C15H16O4 243.0959 [M + H–H2O]+ [19]
261.1069 [M + H]+
(3.2) 215.1020 [M + H–2H2O]+
12 23.295 Jacquinelin (4) 285.1026 [M + Na]+ C15H18O4 245.1120 [M + H–H2O]+ [20]
263.1222 [M + H]+ 217.1171 [M + H–2H2O]+
(−2.6)
13 26.852 11β,13-Dihydrolactucopicrin (5) 435.1318 [M + Na]+ C23H24O7 261.1939 [M + H–hydroxyphe- [17]
nyl acetoxy]+
413.1504 [M + H]+
(0.8)

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 Eur Food Res Technol

Table 1  continued
Peak No. T (min) Identification MS (error, ppm) Formula MS/MS (m/z) References
+
14 26.959 Lactucopicrin (6) 411.1348 [M + H] C23H22O7 259.0908 [M + H–hydroxyphe- [17]
nyl acetoxy]+
409.1379 [M − H]−
(1.9)
15 28.177 Erypoegin F 353.2217 [M + H]+ C21H20O5 335.3115 [M + H–H2O]+ [21]
(3.6)
16 30.886 Scopoletin 193.0811 [M + H]+ C10H8O4 175.0728 [M + H–H2O]+ [22]
191.0772 [M − H]−
(5.8)
17 31.158 Magnolialide 249.2117 [M + H]+ C15H20O3 201.1978 [M + H–H2O–2CH3]+ [23]
(2.6)
18 32.985 Caftaric acid 313.2291 [M + H]+ C13H12O9 295.3133 [M + H–H2O]+ [24]
311.1847 [M − H]− 277.300 [M + H–2H2O]+
(−2.1)
19 36.254 3,4-Dihydro-15-dehydrolactu- 411.3772 [M + H]+ C23H22O7 393.4270 [M + H–H2O]+ [17]
copicrin
(5.01)
20 38.65 Myricetin 319.3925 [M + H]+ C15H10O8 283.3638 [M + H–2H2O]+ [25]
(−3.7)
21 40.231 Crepidiaside A 423.2625 [M + H]+ (4.8) C21H26O9 261.2316 [M + H–Glc]+ [26]
22 42.668 Not identified 537.2900 [M + Na]+ C23H46O12 353.2604 [M + H–Glc]+
515.3080 [M + H]+ 335.3486 [M + H–Glc–H2O]+
(3.68)
23 45.319 Isochlorogenic acid C 539.3067 [M + Na]+ C25H24O12 355.2753 [M + H–caffeoyl]+, [27]
517.3115 [M + H]+ 337.2651 [M + H–caffeoyl–
H2O]+
(2.78)
24 46.175 Isochlorogenic acid A 539.3067 [M + Na]+ C25H24O12 355.2762 [M + H–caffeoyl]+, [27]
517.3119 [M + H]+ 337.2658 [M + H–caffeoyl–
H2O]+
(2.46)
25 51.527 Hieracin II 301.1327 [M + Na]+ C15H18O5 261.2504[M + H–H2O]+ [28]
279.1516 [M + H]+ (4.5)
26 56.7 Lactupicrin methyl ester 443.477 [M + H]+ C24H26O8 425.3717 [M + H–H2O]+ [29]
(−3.6) 396.4067 [M + H–H2O–CO]+
27 59.315 β-Sitosterol 415.7131 [M + H]+ (2.9) C29H50O 399.6870 [M + H–H2O]+ [30]
28 61.259 Cyanidin chloride 345.6310 [M + Na]+ C15H11ClO6 287.2410 [M + H–Cl]+ [31]
323.6493 [M + H]+
(−4.25)

olefinic carbons at δ 175.14 (C-4), 148.49 (C-10), 132.20
(C-3) and 131.73 (C-1), one methylene at δ 48.28 (C-9),
four oxygen-bearing methines at δ 81.05 (C-6), 68.60
(C-15), 61.73 (C-8), 60.86 (C-7) and two methyls at δ
20.44 (C-14) and 14.41 (C-13) revealed the presence of
three rings and further deduced as lactone-type sesquiterpe-
noid [32]. Moreover, the approval information from its 1H
NMR spectrum in accordance with literature could identify
compound 1 as 11β,13-dihydrolactucin (1) [18].
Compounds 2–6 were all isolated as white powder. All of
the present signals in the NMR spectra were similar as those
of compound 1, suggesting that they were lactone-type ses-
quiterpenoids. By comparison of the chemical shift with
literature data, the compounds were identified as lactucin
(2) [33], 8-deoxylactucin (3) [19], jacquinelin (4) [34, 35],
11β,13-dihydrolactucopicrin (5) [36], lactucopicrin (6) [33],
respectively. Their structure and the 13C NMR data of these
compounds are presented in Fig. 6 and Table 2, respectively.

13
Eur Food Res Technol

Fig. 6  Structure of compounds
1–6

1: R = OH 2: R = OH
4: R = H 3: R = H
5: R = A 6: R = A

Table 2  13C NMR data of Position 1 2 3 4 5 6

compounds 1–6 (d in ppm)
1 131.7 131.8 131.8 131.7 131.8 131.1
2 195.9 195.8 196.3 196.0 195.5 195.4
3 132.2 132.8 134.9 135.2 132.8 132.0
4 175.1 174.9 169.3 169.5 175.0 174.7
5 48.6 48.2 52.0 49.7 47.2 47.1
6 81.1 81.3 84.3 84.2 80.7 81.0
7 57.8 57.0 55.1 55.4 53.1 54.0
8 61.7 67.8 25.3 25.9 70.8 69.6
9 48.2 47.2 36.7 36.8 57.8 43.5
10 148.5 149.0 152.4 152.8 147.0 146.5
11 41.1 137.8 139.2 40.8 40.0 136.5
12 178.5 169.4 169.5 178.4 177.6 168.7
13 14.4 121.8 117.8 11.8 13.8 120.7
14 20.4 20.4 23.1 20.43 20.0 20.4
15 62.6 61.7 61.9 62.7 61.0 61.67
1′ 124.3 124.2
2′ 130.0 130.1
3′ 114.9 115.1
4′ 156.4 156.5
α 40.8 40.2
β 171.1 171.0

Bioactivity of extracts and compounds Table 3  Inhibitory effect of six compounds isolated from chicory
roots against α-glucosidase. Acarbose was used as a positive control
The enzymatic assay showed that EAFE yielded the high- Compounds α-Glucosidase (IC50)
est α-glucosidase inhibition among the three crude extracts
11β-13-Dihydrolactucin (1) 17.46 ± 0.125
(Fig. 2), and Fr. C yielded the highest α-glucosidase inhibi-
Lactucin (2) 14.71 ± 0.221
tion among the six subfractions Fr. A–Fr. F from EAFE.
8-Deoxylactucin (3) 15.02 ± 0.513
The α-glucosidase inhibition activity of those com-
pounds isolated from subfraction Fr. C is shown in Jacquinelin (4) 4.180 ± 0.128
Table  3. The current data suggested that all six com- 11β,13-Dihydrolactucopicrin (5) 27.49 ± 0.17
pounds had inhibitory activity against α-glucosidase, Lactucopicrin (6) 14.71 ± 0.117
and their inhibitory concentration 50 (IC50) val- Acarbose 6.353 ± 0.228
ues were 17.46 ± 0.125 μM (1), 14.71 ± 0.221 μM

13

(2), 15.02 ± 0.513 μM (3), 4.180 ± 0.128 μM (4),
27.49 ± 0.17 μM (5), 14.71 ± 0.117 μM (6), respectively.
In particular, compound 4 showed lower IC50 value lower
than that of acarbose (6.353 ± 0.228 μM), implying that
compound 4 has outstanding inhibitory activities against
yeast α-glucosidase. It would be worthy to further investi-
gate the anti-diabetic effect in appropriate studies.
Sesquiterpene lactones constitute a wide range of bio-
logically active plant chemicals from many species of
medicinal plants [37]. Their structural diversity and diverse
potential biological activities such as anti-inflammatory,
anticancer, anti-bacterial, antifungal and their potential
anti-diabetic activity are far less explored [38]. The pre-
sent biological screening suggests that sesquiterpenes in
chicory seem more active as inhibitor of the enzyme stud-
ied. Our results show that all compounds revealed a signif-
icant potency with IC50 values lower than 50 μM, which
may be owing to their similar structure of a ɤ-lactone ring
(closed to a 5/7-bicyclic) [32]. By comparison of the dem-
onstrated α-glucosidase activities of identified compounds,
compound 5 showed limited inhibition activity, suggesting
that phenyl group attached to lactone-type sesquiterpe-
noid will decrease its ability, as demonstrated in this study,
which might due to its structure become harder to bond
α-glucosidase [39].
The employed bioactivity-oriented approach can suc-
cessfully isolate and purify the main bioactive compounds
from chicory roots for α-glucosidase inhibition. Successive
fractionation might cause degradation of the active con-
stituents, different compounds interacted with each other,
leading to synergistic or antagonistic effects [40]. In the
present research, the inhibitory activity of EAFE against
α-glucosidase was stronger than either the subsequent Fr.
C or any single compound at same concentration, proven
the presence of possible synergistic effect of sesquiterpene
compounds.
Conclusions
Diabetes mellitus (DM) is a common metabolic disorder
characterized by hyperglycemia, with symptoms of fre-
quent urination, increased thirst and increased hunger [41].
As well known, many studies have been carried out and
showed that synthetic α-glycosidase inhibitors have many
undesirable adverse effects, especially gastrointestinal
side effects [42]. Consequently, surveys have concentrated
on discovering effective and safe α-glycosidase inhibitors
from natural products [43]. In this study, the bioassay-
guided isolation was used to evaluate α-glycosidase inhibi-
tory activity in search of the responsible active components
of chicory roots. The detection of chemical composition
of the fraction with high α-glycosidase inhibition showed
13
Eur Food Res Technol
that fourteen compounds among twenty-eight ones were
sesquiterpenes or sesquiterpene glycosides, implying that
the sesquiterpenes may be the main ingredients for the
inhibitory activity of chicory root against α-glucosidase.
Further bioassay-guided isolation yielded six compounds,
and it was found that jacquinelin (4) had the strongest
α-glucosidase inhibition activity. These results provide
basic information for further research on utilizing chicory
as natural anti-diabetic resource.
Acknowledgements This study was financially supported by the
grant from Jiangsu Scientific and Technological Innovations Platform
(No. BM2011117).
Compliance with ethical standards 
Conflict of interest None.
Compliance with ethics requirements  This article does not contain
any studies with human or animal subjects.
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