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Butterfly pea ( Clitoria ternatea ) seed and petal extracts decreased HEp-2 carcinoma cell viability

  • the south subtropical crops research institute, Chinese Academy of Tropical Agricultural Science (CATAS), Zhanjiang

Abstract and Figures

The hydrophilic phenolics, lipophilic tocopherols, phytosterols and fatty acids in butterfly pea seeds and petals were determined. The seeds had fifteen phenolics; of them, sinapic acid, epicatechin and hydroxycinnamic acid derivative concentrations were above 0.5 mg g-1. The petals contained a group of ternatins, flavone glycosides and delphinidin derivatives. Both the seeds and petals had four phytosterols and α- and γ-tocopherols. However, the level of β-sitosterol or γ-tocopherol in the seeds was much higher than in the petals. Linoleic acid was the most abundant fatty acid in the seeds and petals, while phytanic acid was found in the petals. The effect of lipophilic and hydrophilic extracts of the seeds [lipophilic extract of the butterfly pea seeds (LBS) and hydrophilic extract of butterfly pea seeds (HBS)] and petals [lipophilic extract of the butterfly pea petals (LBP) and hydrophilic extract of butterfly pea petals (HBP)] on decreased HEp-2 human carcinoma cell viability was evaluated. The effect of HBS or HBP on decreased cancer cell viability was much higher than that of either LBS or LBP, while HBS showed significantly higher effect than HBP. The results indicated that butterfly pea seed and petal extracts could have the potential in functional food development. International Journal of Food Science and Technology
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Original article
Butterfly pea (Clitoria ternatea) seed and petal extracts decreased
HEp-2 carcinoma cell viability
Yixiao Shen,
Liqing Du,
Haiying Zeng,
Xiumei Zhang,
Witoon Prinyawiwatkul,
Jose R. Alonso-Marenco
Zhimin Xu
1 School of Nutrition and Food Sciences, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA
2 The Key Laboratory of Tropical Fruit Biology of Ministry of Agriculture, The South Subtropical Crop Research Institute, Chinese
Academy of Tropical Agricultural Science, 20 Jiefang W Rd, Zhanjiang, Guangdong 524001, China
3 School of Liquor and Food Engineering, Guizhou University, Xueshi Rd, Huaxi District, Guiyang, Guizhou 550025, China
(Received 23 February 2016; Accepted in revised form 29 April 2016)
Summary The hydrophilic phenolics, lipophilic tocopherols, phytosterols and fatty acids in butterfly pea seeds and
petals were determined. The seeds had fifteen phenolics; of them, sinapic acid, epicatechin and hydrox-
ycinnamic acid derivative concentrations were above 0.5 mg g
. The petals contained a group of ter-
natins, flavone glycosides and delphinidin derivatives. Both the seeds and petals had four phytosterols
and a- and c-tocopherols. However, the level of b-sitosterol or c-tocopherol in the seeds was much higher
than in the petals. Linoleic acid was the most abundant fatty acid in the seeds and petals, while phytanic
acid was found in the petals. The effect of lipophilic and hydrophilic extracts of the seeds [lipophilic
extract of the butterfly pea seeds (LBS) and hydrophilic extract of butterfly pea seeds (HBS)] and petals
[lipophilic extract of the butterfly pea petals (LBP) and hydrophilic extract of butterfly pea petals (HBP)]
on decreased HEp-2 human carcinoma cell viability was evaluated. The effect of HBS or HBP on
decreased cancer cell viability was much higher than that of either LBS or LBP, while HBS showed signif-
icantly higher effect than HBP. The results indicated that butterfly pea seed and petal extracts could have
the potential in functional food development.
Keywords Cell viability, lipids, phytosterols, polyphenols, ternatins, tocopherols.
Butterfly pea (Clitoria ternatea), a member of Faba-
ceae family and Papilionaceae subfamily, is a perennial
leguminous twiner. Approximately 60 butterfly pea
species are distributed within the tropical belt, while a
few species are found in temperate areas (Al-Asmari
et al., 2014). The anthocyanins abundant in butterfly
pea petals make the petals as a natural blue colorant
source for a variety of food products (Mukherjee
et al., 2008). Also, triterpenoids, flavonol glycosides
and alkaloids were found in butterfly pea leaves, while
pentacyclic triterpenoids, taraxerol and taraxerone
were identified in the roots (Singh & Tiwari, 2010).
However, the chemical composition of butterfly pea
seeds, especially its bioactive constituents such as phe-
nolics, tocols and phytosterols, has not been docu-
mented. In this study, nonpolar solvent and polar
solvent were used to obtain lipophilic and hydrophilic
extracts of butterfly pea seeds and petals, respectively.
The phytochemicals in each extract were identified for
exploring the potential bioactive components in the
butterfly pea seeds and petals. Those identified phyto-
chemicals are a group of antioxidants that may play
an important role in the bioactive functions of butter-
fly pea.
The reported health-promoting functions of butterfly
pea include antidiabetic, nootropic, anxiolytic, anticon-
vulsant, sedative, antipyretic, anti-inflammatory and
analgesic functions (Jain & Shukla, 2011). However,
the anticancer potential of butterfly pea has not been
studied. In this study, a laryngeal carcinoma cell line
was used to examine the anticancer effect of butterfly
pea seed and petal extracts. Laryngeal carcinoma
accounts for 25% of head and neck carcinoma and is
the second most common respiratory tract cancer fol-
lowing lung cancer (Mirunalini et al., 2011). The limit-
less replication of the cancer cells and the multiple
interactions with their microenvironments increase the
*Correspondent: Fax: 225-578-5300; e-mail:
The first two authors contributed equally to this article.
International Journal of Food Science and Technology 2016, 51, 1860–1868
©2016 Institute of Food Science and Technology
difficulty in treating the cancer (Pienta et al., 2008).
Primary clinical treatments for laryngeal cancer are
surgery, chemotherapy and radiotherapy. However,
they induce seriously adverse side effects or even result
in resistance to these therapies (Agostinis et al., 2011).
Recently, much attention to cancer treatment has been
focused on some plant-derived compounds that have
pharmacological functions on the tumour but have less
side effects (Veerabadran et al., 2013). In this study,
the anticancer effect of hydrophilic and lipophilic
extracts of butterfly pea petal and seed on decreased
laryngeal cancer cell (HEp-2) viability was studied and
compared. The aim of this study was to provide the
bioactive phytochemical profiles in both butterfly pea
petals and seeds and also emphasise the role of those
bioactive compounds in anticancer activity.
Materials and methods
Chemicals and materials
High-performance liquid chromatography (HPLC)-
grade acetonitrile, acetic acid, methanol and hexane
were purchased from Fisher Chemicals (Fair Lawn,
NJ, USA). Acetone was purchased from Macron
(Charlotte, NC, USA). Ethyl acetate was purchased
from EM Science (Gibbstown, NJ, USA). Trimethylsi-
lyl imidazole (TMS), BCl
methanol and phenolics,
fatty acids and tocopherol standards were purchased
from Sigma Aldrich (St. Louis, MO, USA). Fresh but-
terfly pea seeds and petals (Clitoria ternatea) were
obtained from a local garden in Baton Rouge, LA,
USA. The human carcinoma HEp-2 cell line was pur-
chased from American Type Culture Collection
(ATCC, Manassas, VA, USA). Other reagents and
culture media containing foetal bovine serum (FBS),
antibiotic (penicillinstreptomycin), CellTiter-Blue,
dimethyl sulphoxide (DMSO) and phosphate-buffered
saline (PBS) were purchased from Invitrogen (Grand
Island, NY, USA).
Extraction of hydrophilic and lipophilic bioactive
compounds in butterfly pea seeds and petals
The freeze-dried butterfly pea seeds and petals were
ground by a coffee blender (Hamilton Beach, Southern
Pines, NC, USA). The hydrophilic compounds in the
petals or seeds (20 g) were extracted using 50 mL of
methanol at 60 °C for 20 min. After centrifugation,
the methanol layer was transferred to a clean tube.
Then, the solid residues were extracted repeatedly for
two more times at the same condition and using the
same procedure. The dried hydrophilic extract was
obtained by evaporating the collected methanol using
a vacuum centrifuge evaporator (Labconco, Kansas
City, MO, USA). The dried extracts were then
prepared for a stock solution (50 mg mL
in metha-
nol). For the lipophilic extract of butterfly pea petals
or seeds, 50 mL of a solvent mixture of ethyl acetate
and hexane (50:50; v:v) was used for the extraction
according to the same procedure used for obtaining
the hydrophilic extract. After the solvent extract was
dried, a stock solution of the lipophilic extract
(50 mg mL
in hexane) was prepared as well.
Identification and quantification of hydrophilic and
lipophilic phytochemicals and fatty acids
Phytochemical profiles of the hydrophilic extracts were
determined by a reverse-phase HPLC (2690; Waters,
Torrance, CA, USA) coupled with C18 column (id
250 94.60 mm, 5 lm; Phenomenex, Torrance, CA,
USA) and a diode array detector and the operation
condition for HPLC was as reported in the study of
Du et al. (2014). The concentrations of the phenolics
were calculated based on their corresponding standard
curves. Both the extracts were also subject to an LC-
MS analysis to identify the compounds without their
standards available. The LC-MS consisted of a ultra
performance liquid chromatography (UPLC) system
(Thermo Scientific Dionex UltiMate 3000, Waltham,
MA, USA) and a mass spectrometry (MS) (Q Exac-
Plus Hybrid Quadrupole-Orbitrap
) that had
an electrospray ionisation source (ESI) in the positive
mode with a full MS scan from 150 to 2000 m/z. The
LC-MS separation was carried out using a reverse-
phase column (Acclaim
Mixed-Mode WAX-1,
150 92.1 mm, 5 lm). The mobile phase consisted of
solution A (1% formic acid solution) and solution B
(acetonitrile) at a constant flow rate of 0.2 mL min
with a gradient programme of 070% B at 0.0
8.0 min, 70100% B at 8.010.0 min, 1000% B at
1015 min. The MS parameters were set as follows:
electric potential of the ESI source, 3.0 kV; capillary
temperature, 300 °C; heater temperature, 200 °C. The
concentrations of ternatins and delphinidin derivative
were calculated by the standard curve of cyanidin
chloride in molar concentration and then converted to
mass unit (mg g
) in the sample based on their
molecular weights. Tocopherols in the samples were
determined by a normal-phase HPLC (1100 series;
Agilent, Santa Clara, CA, USA) with Supelcosil LC-Si
column (id 250 94.60 mm 5 lm, Supelco, Bellefonte,
PA, USA). The condition of the HPLC analysis was
the same as the method described in Jang & Xu
The fatty acids in each lipophilic extract were esteri-
fied using 2 mL of BCl
(boron trichloride) methanol
after 200 lg of internal standard (C17:0) was added.
The reaction mixture was incubated at 60 °C for
30 min. Then, 1 mL of water and 1 mL of hexane
were added to the reaction solution and vortexed.
©2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al. 1861
After centrifugation, the upper hexane layer was trans-
ferred to a clean test tube and mixed with anhydrous
to remove any moisture before it was trans-
ferred to a gas chromatography (GC) vial. The fatty
acids were determined by a GC equipped with an
flame ionisation detector (FID) and a Supelco SP2380
(30 m 90.25 mm) column. The GC condition was the
same as that used in the study of Yue et al. (2008).
The determination of phytosterols was based on the
study of Xu & Godber (1999). After the lipophilic
extract was mixed with an aliquot of hexane contain-
ing 20 lg cholesterol internal standard, it was dried
and then reacted with 200 lL of TMS and 50 lLof
acetonitrile at 65 °C for 30 min. The derived products
were extracted using 200 lL of hexane and analysed
by GC-MS. A Varian CP-3800 GC (Walnut Creek,
CA, USA) coupled with a Saturn 2200 mass spectrom-
eter and a DB-5 column (60 m 90.25 mm) (Supelco)
was used in the analysis. The initial oven temperature
was 200 °C. Then, it was ramped to 280 °C at a rate
of 10 °C min
and held at the final temperature for
62 min. Helium was the carrier gas at a constant flow
rate of 1.5 mL min
. The injection port temperature
and split ratio were 280 °C and 1:50, respectively. The
ratios of peak areas at different levels (5, 10, 20, 50
and 100 lg) of campesterol, stigmasterol and b-sitos-
terol to 20 lg of cholesterol internal standard were
used to set up the standard curves for quantifying the
Determination of capability of decreasing human
carcinoma cell (HEp-2) viability
The human carcinoma HEp-2 cell line was used to
assess the potential of butterfly pea seed and petal
extracts in decreasing cancer cell viability. The cell line
was cultured in Dulbecco’s modified Eagle’s media
(DMEM), supplemented with 10% foetal bovine
serum (FBS) and 1% antibiotic (penicillinstrepto-
mycin), and grown in a 5% CO
atmosphere with
95% humidity at 37 °C for 24 h. Then, the cells were
harvested, counted (3 910
cells mL
) and trans-
ferred into a 96-well plate. The working solution of
each extract was prepared by dissolving the extract
with 0.2% DMSO in PBS culture media. The treat-
ment solutions were prepared based on a group of the
concentrations multiplied by the low and effective con-
centration of each type of extract that was obtained in
our preliminary study. The HEp-2 cells were incubated
with a series of concentrations of the hydrophilic
extract working solution (1.0, 0.5, 0.25, 0.12 and
0.06 mg mL
) or lipophilic extract working solution
(12.0, 9.0, 6.0, 3.0 and 1.5 mg mL
) for 96 h at
37 °C for a dose-dependent study. The cells only
mixed with the media and 0.2% DMSO were used as
the control group. For measuring the cell viability, the
media were discarded and replaced with 100 lLof
fresh media containing 20% CellTiter-Blue. After the
cells were stained for 4 h, the fluorescence intensity of
the media was read at excitation/emission wavelengths
of 570/615 nm using a FluoStar Optima microplate
reader (BMG, Ortenberg, Germany). The potential of
the extract in decreasing human carcinoma cell viabil-
ity at each concentration treatment was expressed by a
survival rate, which was the percentage of the intensity
of the treatment vs. the intensity of the control.
Data analysis
The determinations of hydrophilic and lipophilic phy-
tochemicals and fatty acids in each extract were
repeated in triplicate and expressed as means stan-
dard deviation. The significant differences between the
concentrations of each compound in the hydrophilic
or lipophilic extracts were determined by one-way
ANOVA at P<0.05 (SAS, 9.1.3, Cary, NY, USA). The
determination of the potential of each treatment con-
centration or control in decreasing the cell viability
was repeated five times and analysed by GraphPad
Prism (version 6.0; GraphPad Software Inc., La Jolla,
CA, USA). The differences in the potential in decreas-
ing the cell viability between the treatments and con-
trol were analysed by two-way ANOVA at P<0.05.
Results and discussions
Hydrophilic and lipophilic phytochemicals and fatty acids
in butterfly pea seeds and petals
The yields of the HBS and HBP extracts were 3.66
and 4.05%, respectively. The chromatograms of the
hydrophilic phenolics in HBS and HBP are shown in
Figs 1 and 2, respectively. In addition to the high con-
centration of ascorbic acid [1.32 0.02 mg g
weight (FW)], sinapic acid was the dominant phenolic
compound among the fifteen major hydrophilic pheno-
lics at a concentration of 1.01 0.07 mg g
FW, fol-
lowed by epicatechin and gallic acid in the seed
(Table 1). Compared with the contents in rapeseeds
(0.090.59 mg g
FW) and camelina seeds
(0.39 mg g
FW) (Ni
c & Abramovi
c, 2014),
the sinapic acid content in butterfly pea seeds was
much higher than each of them (Table 1). It was
reported that sinapic acid could help suppress the
expression of proinflammatory mediators via NF-jB
inactivation in regulating inflammatory status and
immune response (Yun et al., 2008). Epicatechin, in
the butterfly pea seeds, which was reported to exhibit
immunoregulatory, antihypertensive effects as well,
was ten times higher (0.56 mg g
FW, Table 1) than
that reported in the garden pea seeds (Pisum sativum)
(0.05 mg g
) (Ferraro et al., 2014; Litterio et al.,
©2016 Institute of Food Science and TechnologyInternational Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al.1862
2015). Protocatechuic, p-coumaric, rutin and two
hydroxycinnamic acid derivatives in the butterfly pea
seeds were all above 0.30 mg g
FW, while kaemp-
ferol, apigenin, caffeic, syringic, ferulic, rosmarinic and
cinnamic acids were in a range of 0.04 to 0.22 mg g
FW (Table 1). Compared with the rutin content in edi-
ble Amaranthus seeds, which was between 0.0711 and
0.0775 mg g
dry weight (DW) (Li et al., 2015), the
rutin content in butterfly pea seeds was much higher
and reached 0.31 mg g
FW (Table 1). Although
mung bean, radish, broccoli and sunflower seeds con-
tain gallic, protocatechuic, caffeic, p-coumaric, ferulic,
chlorogenic, sinapic acid, quercetin and kaempferol
zket al., 2014), each of their levels was signifi-
cantly lower than that in the butterfly pea seeds.
The unique phytochemicals in butterfly pea petals
are ternatins, a group of polyacylated delphinidin
derivatives (Sasaki et al., 2013). It was reported that
ternatin A1-A3, B1-B4, C1-C5 and D1-D3 consist of
delphinidin 3, 30,5
0-triglucoside attached with malonic
acid, glucose, p-coumaric acid or caffeic acid (Kazuma
et al., 2003). In this study, ternatin C2 and D2 in the
butterfly pea petals were observed at higher concentra-
tions of 1.81 0.09 and 1.45 0.07 mg g
respectively. The concentrations of ternatin A1, B3,
D3 and B2 were in a range of 0.32 to 0.51 mg g
(Table 2). The chemical structures of the six types of
ternatins are elucidated in Fig. 3. Two delphinidin
derivatives and cyanidin-3-sophoroside were also iden-
tified and responsible for the blue colour of the petals
together with ternatins. In general, kaempferol 3-neo-
hesperidoside, quercetin 3-(2G-rhamnosylrutinoside)
and rutin were the major flavanol glycoside com-
pounds in the petals, while ellagic acid was the only
phenolic acid identified in butterfly pea petals
(Table 2).
The yields of the lipophilic extracts, LBS and LBP,
were 5.28 and 0.80%, respectively. Lipophilic phytos-
terols are chemically characterised as triterpenes and
considered to be one of the structural components of
10.00 20.00 30.00 40.00 50.00 60.00 70.00
Figure 1 Chromatogram of the hydrophilic
butterfly pea seed extract. 1 vitamin C; 2
gallic acid; 3 protocatechuic acid; 4 epi-
catechin; 5 caffeic acid; 6 syringic acid;
7sinapic acid; 8 hydroxycinnamic acid
derivatives; 9 p-coumaric acid; 10
hydroxycinnamic acid derivatives; 11 rutin;
12 ferulic acid; 13 rosmarinic acid; 14
cinnamic acid; 15 kaempferol; 16 api-
10.00 20.00 30.00 40.00 50.00 60.00 70.00
2 4
10 12
Figure 2 Chromatogram of the hydrophilic
butterfly pea petal extract. 1 cyanidin-
3-sophoroside; 2 delphinidin derivative; 3
ternatin A1; 4 ternatin B3; 5 ternatin
D3; 6 ellagic acid; 7 rutin; 8 delphini-
din derivative; 9 kaempferol-3-neohesperi-
doside; 10 quercetin-3-(2G-
rhamnosylrutinoside); 11 ternatin B2;
12 ternatin C2; 13 ternatin D2.
©2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al. 1863
plant cell membranes (Moreau et al., 2002). Similar to
the function of cholesterol in animal cells, free phytos-
terols serve to stabilise phospholipid bilayers in the
plant cells (Moreau et al., 2002). Although several
studies investigated phytosterols in Clitoria ternatea
species, most of them only focused on leaves, roots or
petals (Kapoor & Purohit, 2013). In this study, b-sitos-
terol (40.17 3.73 mg/100 g FW) in the butterfly pea
seed extract LBS was significantly higher than that of
the petal extract LBP (6.77 0.19 mg/100 g FW)
(Table 3). It was reported that the level in butterfly
pea roots and shoots was between 6 and 9 mg/100 g
(Kapoor & Purohit, 2013). Also, the seed extract LBS
contained campesterol at a level of 8.07 0.22 mg/
100 g FW, which was several times higher than that of
the petal extract LBP (1.24 0.02 mg/100 g FW)
(Table 3) and some vegetable seeds such as pepper
seeds (4.235.41 mg/100 g FW) and tomato seeds
(1.086.56 mg/100 g FW) (Silva et al., 2013; Ancos
et al., 2015). However, the levels of stigmasterol in
butterfly pea seeds and petals were similar
(7.95 0.63 and 6.70 0.83 mg/100 g FW, respec-
tively) (Table 3) and approximately five and eight
times higher than in berryfruit (0.501.60 mg/100 g
FW) and pepper seeds (0.630.93 mg/100 g FW) (Silva
et al., 2013; Salvador et al., 2015). Phytosterols were
confirmed to possess hypocholesterolaemic function
and reduce the risks of benign prostatic hyperplasia,
cardiovascular diseases, colon and breast cancer devel-
opment, as well as immunological effects in macro-
phages (Hamedi et al., 2014).
As for the tocol contents, the butterfly pea seeds
had abundant c-tocopherol (5.44 0.30 mg/100 g
FW) compared with the grape seeds (14.1
30.2 mg kg
) and Jatropha curcas seeds
(33.9 mg kg
) reported in the studies of Sabir et al.
(2012) and Corzo-Valladares et al. (2012), respectively.
However, c-tocopherol in the butterfly pea petals was
twenty times lower than that in the seeds (Table 3).
The levels of a-tocopherol in butterfly pea seeds and
petals were similar (0.17 0.06 and 0.20 0.06 mg/
100 g FW, respectively) (Table 3). These tocols have
been evidenced for protecting cell membrane against
reactive lipid radicals and for the prevention of
atherosclerosis and carcinogenesis (Yang et al., 2013).
For the fatty acids profile, both the butterfly pea
seeds and petals had palmitic acid (C16:0), stearic acid
(C18:0), petroselinic acid (C18:1), linoleic acid (C18:2),
arachidic acid (C20:0) and behenic acid (C22:0)
(Table 3). Among them, linoleic acid was the most
abundant fatty acid and had 8.73 0.61 and
4.72 0.51 mg g
FW in the butterfly pea seeds and
petals, respectively. It is well known that linoleic acid
is an essential fatty acid and required for assisting nor-
mal biological activities in the brain and heart (Blan-
chard et al., 2013). The palmitic, stearic and
petroselinic acids in the seeds and petals were all above
1.0 mg g
(Table 3). Different from the results of a
previous study (Mukherjee et al., 2008), arachidic and
behenic acids were first time observed in the butterfly
pea by this study (Table 3). Furthermore, phytanic
acid was found in the butterfly pea petals and might
be derived from a microbial breakdown of chlorophyll
to release phytol followed by the further oxidation to
phytanic acid (Jansen & Wanders, 2006). In the study
Table 1 Hydrophilic compounds identified in the butterfly pea
Peak No. Compounds
(mg g
1 Ascorbic acid 1.32 0.02
2 Gallic acid 0.42 0.00
3 Protocatechuic acid 0.34 0.01
4 Epicatechin 0.56 0.03
5 Caffeic acid 0.22 0.01
6 Syringic acid 0.14 0.01
7 Sinapic acid 1.01 0.07
8 Hydroxycinnamic
acid derivative 1
0.57 0.19
9p-Coumaric acid 0.30 0.01
10 Hydroxycinnamic
acid derivative 2
0.44 0.02
11 Rutin 0.31 0.01
12 Ferulic acid 0.15 0.00
13 Rosmarinic acid 0.05 0.00
14 Cinnamic acid 0.08 0.00
15 Kaempferol 0.04 0.00
16 Apigenin 0.09 0.00
Table 2 Hydrophilic compounds identified in the butterfly pea
Peak No. Compounds
(mg g
1 Cyanidin-3-sophoroside 0.31 0.02
2 Delphinidin derivative 0.28 0.01
3 Ternatin A1 0.51 0.03
4 Ternatin B3 0.50 0.03
5 Ternatin D3 0.54 0.01
6 Ellagic acid 0.21 0.01
7 Rutin 0.89 0.04
8 Delphinidin derivative 2.13 0.16
9 Kaempferol
1.76 0.05
10 Quercetin
0.37 0.01
11 Ternatin B2 0.32 0.01
12 Ternatin C2 1.81 0.09
13 Ternatin D2 1.45 0.07
©2016 Institute of Food Science and TechnologyInternational Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al.1864
of Jansen & Wanders (2006), phytanic acid was
involved in several mechanisms for regulating triglyc-
erides/cholesterol status in the skeletal muscles.
Capabilities of hydrophilic and lipophilic butterfly pea
seed and petal extracts in decreasing carcinoma cell
(HEp-2) viability
The media with different concentrations of the four
extracts HBS, HBP, LBS and LBP were prepared,
respectively, and used to treat HEp-2 cells. HBS was
the most effective extract against the survival of
HEp-2 cells which rapidly decreased from 100.0 to
7.2% as the level of HBS increased from 0 to
0.25 mg mL
(Fig. 4). However, the survival rates of
HEp-2 in HBP treatment still remained over 90% in
the same concentration range (Fig. 4). As the concen-
tration of HBP increased from 0.25 to 0.50 mg mL
the survival rate of HEp-2 then rapidly reduced to
17.2% (Fig. 4). Both HBP and HBS could decrease
95% of the HEp-2 cell viability after the concentration
was increased to 1.0 mg mL
(Fig. 4). On the other
hand, the decreasing effect was observed in LBS and
LBP treatment only after their concentrations were
increased to 1.5 mg mL
. There was no significant
difference between the two treatments with their
concentrations lower than 6 mg mL
(Fig. 4). At a
concentration of 9 mg mL
, LBS exhibited approxi-
mately 2.5 times higher capability than LBP in
decreasing HEp-2 cell growth. In general, compared
with the lipophilic extracts of seeds and petals, the
hydrophilic extracts had much higher capability in
decreasing carcinoma viability (Fig. 4).
Generally, Krebs cycle is the primary metabolic
pathway for providing ATP for normal cell growth
through the involvement of mitochondria (Wen et al.,
2013). Different from the normal cells, however, can-
cer cells inevitably rely on metabolic reprogramming
and undergo glycolytic pathway to carry out a rapid
energy generation and macromolecular synthesis
because of mitochondria dysfunction (Suh et al.,
2013). The high glycolytic fluxes in cancer cells fur-
ther induce apoptosis in the neighbourhood normal
cells, block immune system and induce tissue inva-
sion by tumours (Suh et al., 2013). Thus, the inhibi-
tion of glycolysis is a biochemical way for decreasing
the cancer cell viability with the minimal residual sys-
temic toxicity and can be used in designing therapeu-
tic strategies. Because the butterfly pea seeds
contained abundant phenolics, they might act individ-
ually or synergistically to deactivate the key enzyme
involved in glycolytic metabolism in the cancer cells
D3 D2
Figure 3 Chemical structures of ternatin
A1, B2, B3, C2, D2 and D3.
Table 3 Lipophilic compounds identified in the butterfly pea seeds
and petals
Compounds Seeds Petals
Fatty Acid
(mg g
Palmitic acid
3.61 0.13b 2.13 0.18a
Stearic acid
2.85 0.15b 1.99 0.16a
1.55 0.10b 1.01 0.04a
Linoleic acid
8.73 0.61b 4.72 0.51a
Arachidic acid
0.46 0.03b 0.36 0.01a
Behenic acid
0.41 0.02b 0.30 0.03a
Phytanic acid N.D. 0.81 0.06a
(mg/100 g FW)
Campesterol 8.07 0.22b 1.24 0.02a
Stigmasterol 7.95 0.63a 6.70 0.83a
b-Sitosterol 40.17 3.73b 6.77 0.19a
Sitostanol 5.10 0.05b 1.20 0.03a
(mg/100 g FW)
a-Tocopherol 0.17 0.06a 0.20 0.01a
c-Tocopherol 5.44 0.30b 0.24 0.02a
N.D., not detected
Concentrations in each row with different letters are statistically differ-
ent at p<0.05.
©2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al. 1865
and decrease their viability. It has been reported that
the noncovalent interactions between cellular proteins
and phenolics could prevent the cancer cell prolifera-
tion (Aslan et al., 2015). Phenolics such as rutin,
quercetin, kaempferol, catechin, p-coumaric, sinapic,
ferulic, syringic, caffeic and gallic acids found in the
butterfly pea seeds have also been proved to affect
important control point enzyme (pyruvate kinase
isoenzyme M2) or attack the glucose transporters
(GLUT) in glycolytic pathway to regulate the cancer
cell production (Aslan et al., 2015). Those phyto-
chemicals could also act as apoptosis-inducing factors
and could be released into the cytosol and translo-
cated to the nucleus to cleave DNA (Lee et al.,
2010). Moreover, the other mechanisms of cell cycle
regulation modulation, invasiveness and angiogenesis
suppression would be involved to decrease the HEp2
cancer cell viability (Lee et al., 2010). Thus, it may
be the reason that HBS exhibited the highest effi-
ciency among the four types of extracts in decreasing
HEp-2 cell viability.
Furthermore, different anthocyanins such as ter-
natins and cyanidin glycoside in HBP could be the
compounds for contributing to its anticancer effect as
well. The anthocyanins extracted from black raspber-
ries were found to counteract cancer cell motility
through the disruption of an essential mediator
cyclooxygenase-2 (COX-2) in tumorigenesis (Wang &
Stoner, 2008). However, Dai et al. (2009) suggested
that anthocyanin extract alone could less contribute to
anticancer ability, but may act additively or synergisti-
cally with other active components in the inhibition of
cancer cell growth. It may be the reason that HBS had
greater anticancer potential than HBP.
Tocopherols and phytosterols were the primary
compounds in LBP and LBS. As reported by Kannap-
pan et al. (2012), the anticancer actions of c-toco-
pherol involved in death receptor 5 (DR5) protein
upregulation could further stimulate tumour necrosis
and restrict its proliferation. Of the four phytosterols,
b-sitosterol has been evidenced as the most effective
one in decreasing the growth of cancer cells via the
activation of certain enzymes, which in turn induce
cellular apoptosis (Bradford & Awad, 2007). Woyengo
et al. (2009) suggested that b-sitosterol and campes-
terol could alleviate the cancer development by reduc-
ing the production of carcinogens in biological
metabolism. The levels of b-sitosterol and campesterol
in LBS were approximately seven and eight times
higher than those in LBP, respectively. Thus, it may
explain why LBS had better performance than LBP in
decreasing HEp-2 cell growth in the concentrations
ranging from 5 to 12 mg mL
As discussed above, certain phenolics, tocopherols
and phytosterols could demonstrate the decreasing effi-
ciency by targeting specific enzymes or interfering with
the metabolic pathway of cancer cells without affecting
other nontumorigenic counterparts and cells. Similar
results showed that normal human epidermal ker-
atinocytes and astrocytes were able to survive when
exposed to flavonoids or green tea leaves rich in
polyphenols, while they elicited the death of tumour
cells (Hsu et al., 2003; Das et al., 2010). Therefore, the
butterfly pea seed or petal extracts, especially the
hydrophilic seed extract, have the potential in reducing
the risks of cancer. Although butterfly pea seeds and
flowers have been used as tea or a source of edible
food ingredient for a long time, the possible toxicity
level of the extracts on noncarcinogenic cells should be
considered and evaluated.
The concentration and profile of phenolics, toco-
pherols, phytosterols and fatty acids in butterfly pea
seeds and petals were determined. Anticancer effect of
butterfly pea seed and petal extracts (HBP, HBS, LBP
0.0 0.2 0.4 0.6 0.8 1.0
Concentration of the extracts (mg mL–1)
Concentration of the extracts (mg mL–1)
Survival rate (%)
Survival rate (%)
0 5 10 15
Figure 4 The survival rates of HEp2 cells treated by different con-
centrations of the hydrophilic (HBS and HBP) and lipophilic (LBS
and LBP) extracts of butterfly pea seeds and petals.
©2016 Institute of Food Science and TechnologyInternational Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al.1866
and LBS) was evaluated using HEp-2 carcinoma cell
line. The results indicated that HBS had the highest
capability in decreasing the survival of HEp-2 cells.
Both HBS and HBP exhibited greater performance
than LBS and LBP in decreasing the cell viability. In
general, the hydrophilic extracts of butterfly pea
seed and petal possess a variety of antioxidant phyto-
chemicals and the potential in decreasing cancer cell
We thank Dr. Graca Vicente (Department of Chem-
istry, Louisiana State University) who kindly provided
the facility for the cell culture experiment.
Agostinis, P., Berg, K., Cengel, K.A. et al. (2011). Photodynamic
therapy of cancer: an update. CA: A Cancer Journal for Clinicians,
61, 250281.
Al-Asmari, A.K., Al-Elaiwi, A.M., Athar, M.T., Tariq, M., Al Eid,
A. & Al-Asmary, S.M. (2014). A review of hepatoprotective plants
used in Saudi traditional medicine. Evidence-Based Complementary
and Alternative Medicine,2014, 22.
Ancos, B.De., Collina-Coca, C., Gonzalez-Pena, D. & Sanchez-Mor-
ena, C. (2015). Bioactive compounds from diverse plant, microbial,
and marine sources. In: Biotechnology of Bioactive Compounds:
Sources and Applications (edited by Gupta V.K., Tuohy M.G.,
O’Donovan A. & Lohani M.). Pp. 1415. West Sussex, UK: John
Wiley & Sons.
Aslan, E., Guler, C. & Adem, S. (2015). In vitro effects of some fla-
vonoids and phenolic acids on human pyruvate kinase isoenzyme
M2. Journal of Enzyme Inhibition and Medicinal Chemistry,31,
Blanchard, H., P
edrono, F., Boulier-Month
ean, N., Catheline, D.,
Rioux, V. & Legrand, P. (2013). Comparative effects of well-
balanced diets enriched in a-linolenic or linoleic acids on LC-
PUFA metabolism in rat tissues. Prostaglandins, Leukotrienes and
Essential Fatty Acids (PLEFA),88, 383389.
Bradford, P.G. & Awad, A.B. (2007). Phytosterols as anticancer
compounds. Molecular Nutrition & Food Research,51, 161170.
Corzo-Valladares, P.A., Fern
ınez, J.M. & Velasco, L.
(2012). Tocochromanol content and composition in Jatropha cur-
cas seeds. Industrial Crops and Products,36, 304307.
Dai, J., Gupte, A., Gates, L. & Mumper, R.J. (2009). A compre-
hensive study of anthocyanin-containing extracts from selected
blackberry cultivars: extraction methods, stability, anticancer
properties and mechanisms. Food and Chemical Toxicology,47,
Das, A., Banik, N.L. & Ray, S.K. (2010). Flavonoids activated cas-
pases for apoptosis in human glioblastoma T98G and U87MG
cells but not in human normal astrocytes. Cancer,116, 164176.
Du, L., Shen, Y., Zhang, X., Prinyawiwatkul, W. & Xu, Z. (2014).
Antioxidant-rich phytochemicals in miracle berry (Synsepalum dul-
cificum) and antioxidant activity of its extracts. Food Chemistry,
153, 279284.
Ferraro, K., Jin, A.L., Nguyen, T.D., Reinecke, D.M., Ozga, J.A. &
Ro, D.K. (2014). Characterization of proanthocyanidin metabolism
in pea (Pisum sativum) seeds. BMC Plant Biology,14, 238.
Hamedi, A., Ghanbari, A., Saeidi, V., Razavipour, R. & Azari, H.
(2014). Effects of b-sitosterol oral administration on the prolifera-
tion and differentiation of neural stem cells. Journal of Functional
Foods,8, 252258.
Hsu, S., Bollag, W.B., Lewis, J. et al. (2003). Green tea polyphenols
induce differentiation and proliferation in epidermal keratinocytes.
Journal of Pharmacology and Experimental Therapeutics,306,29
Jain, R.A. & Shukla, S.H. (2011). Pharmacognostic evaluation and
phytochemical studies on stem of Clitoria ternatea linn. Pharma-
cognosy Journal,3,6266.
Jang, S. & Xu, Z. (2009). Lipophilic and hydrophilic antioxidants
and their antioxidant activities in purple rice bran. Journal of Agri-
cultural and Food Chemistry,57, 858862.
Jansen, G.A. & Wanders, R.J.A. (2006). Alpha-oxidation. Biochim-
ica et Biophysica Acta (BBA) Molecular Cell Research,1763,
Kannappan, R., Gupta, S.C., Kim, J.H. & Aggarwal, B.B. (2012).
Tocotrienols fight cancer by targeting multiple cell signaling path-
ways. Genes & Nutrition,7,4352.
Kapoor, B.B.S. & Purohit, V. (2013). Sterol contents from some
fabaceous medicinal plants of Rajasthan desert. Indian Journal of
Pharmaceutical and Biological Research,1,1315.
Kazuma, K., Noda, N. & Suzuki, M. (2003). Flavonoid composition
related to petal color in different lines of Clitoria ternatea.Phyto-
chemistry,64, 11331139.
Lee, J.H., Jeong, Y.J., Lee, S.W. et al. (2010). EGCG induces apop-
tosis in human laryngeal epidermoid carcinoma Hep2 cells via
mitochondria with the release of apoptosis-inducing factor and
endonuclease G. Cancer Letters,290,6875.
Li, H., Deng, Z., Liu, R., et al. (2015). Characterization of pheno-
lics, betacyanins and antioxidant activities of the seed, leaf, sprout,
flower and stalk extracts of three Amaranthus species. Journal of
Food Composition and Analysis,37, 75–81.
Litterio, M.C., Vazquez Prieto, M.A., Adamo, A.M. et al. (2015).
()-Epicatechin reduces blood pressure increase in high-fructose-
fed rats: effects on the determinants of nitric oxide bioavailability.
The Journal of Nutritional Biochemistry,26, 745751.
Mirunalini, S., Arulmozhi, V. & Shahira, R. (2011). Effect of dios-
genin a plant steroid on lipid peroxidation and antioxidant status
in human laryngeal carcinoma cells (Hep2). International Journal
of Pharmacy & Pharmaceutical Sciences,3,94100.
Moreau, R.A., Whitaker, B.D. & Hicks, K.B. (2002). Phytosterols,
phytostanols, and their conjugates in foods: structural diversity,
quantitative analysis, and health-promoting uses. Progress in Lipid
Research,41, 457500.
Mukherjee, P.K., Kumar, V., Kumar, N.S. & Heinrich, M. (2008).
The Ayurvedic medicine Clitoria ternatea From traditional use to
scientific assessment. Journal of Ethnopharmacology,120, 291301.
c, N. & Abramovi
c, H. (2014). Sinapic acid and its deriva-
tives: natural sources and bioactivity. Comprehensive Reviews in
Food Science and Food Safety,13,3451.
zk, P., Socha, R., Gałkowska, D., Ro_
znowski, J. & Fortuna, T.
(2014). Phenolic profile and antioxidant activity in selected seeds
and sprouts. Food Chemistry,143, 300306.
Pienta, K.J., McGregor, N., Axelrod, R. & Axelrod, D.E. (2008).
Ecological therapy for cancer: defining tumors using an ecosystem
paradigm suggests new opportunities for novel cancer treatments.
Translational Oncology,1, 158164.
Sabir, A., Unver, A. & Kara, Z. (2012). The fatty acid and toco-
pherol constituents of the seed oil extracted from 21 grape varieties
(Vitis spp.). Journal of the Science of Food and Agriculture,92,
A.C., Rocha, S.M. & Silvestre, A.J.D. (2015). Lipophilic
phytochemicals from elderberries (Sambucus nigra L.): influence of
ripening, cultivar and season. Industrial Crops and Products,71,
Sasaki, N., Matsuba, Y., Abe, Y. et al. (2013). Recent advances in
understanding the anthocyanin modification steps in carnation
flowers. Scientia Horticulturae,163,3745.
Silva, L.R., Azevedo, J., Pereira, M.J., Valent~
ao, P. & Andrade, P.B.
(2013). Chemical assessment and antioxidant capacity of pepper
©2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al. 1867
(Capsicum annuum L.) seeds. Food and Chemical Toxicology,53,
Singh, J. & Tiwari, K.N. (2010). High-frequency in vitro multiplica-
tion system for commercial propagation of pharmaceutically
important Clitoria ternatea L.A valuable medicinal plant. Indus-
trial Crops and Products,32, 534538.
Suh, D.H., Kim, M.K., Kim, H.S., Chung, H.H. & Song, Y.S.
(2013). Cancer-specific therapeutic potential of resveratrol: meta-
bolic approach against hallmarks of cancer. Future Medicinal
Veerabadran, U., Venkatraman, A., Souprayane, A. et al. (2013).
Evaluation of antioxidant potential of leaves of Leonotis nepetifolia
and its inhibitory effect on MCF7 and HEp2 cancer cell lines.
Asian Pacific Journal of Tropical Disease,3, 103110.
Wang, L.S. & Stoner, G.D. (2008). Anthocyanins and their role in
cancer prevention. Cancer Letters,269, 281290.
Wen, S., Zhu, D. & Huang, P. (2013). Targeting cancer cell mito-
chondria as a therapeutic approach. Future Medicinal Chemistry,5,
Woyengo, T.A., Ramprasath, V.R. & Jones, P.J. (2009). Anticancer
effects of phytosterols. European Journal of Clinical Nutrition,63,
Xu, Z. & Godber, J.S. (1999). Purification and identification of com-
ponents of gamma-oryzanol in rice bran Oil. Journal of Agricul-
tural and Food Chemistry,47, 27242728.
Yang, C.S., Li, G., Yang, Z., Guan, F., Chen, A. & Ju, J. (2013).
Cancer prevention by tocopherols and tea polyphenols. Cancer
Yue, X., Xu, Z., Prinyawiwatkul, W., Losso, J.N., King, J.M. &
Godber, J.S. (2008). Comparison of soybean oils, gum, and defat-
ted soy flour extract in stabilizing menhaden oil during heating.
Journal of Food Science,73, C19C23.
Yun, K.J., Koh, D.J., Kim, S.H. et al. (2008). Anti-inflammatory
effects of sinapic acid through the suppression of inducible nitric
oxide synthase, cyclooxygase-2, and proinflammatory cytokines
expressions via nuclear factor-kappaB inactivation. Journal of Agri-
cultural and Food Chemistry,56, 1026510272.
©2016 Institute of Food Science and TechnologyInternational Journal of Food Science and Technology 2016
Butterfly pea extracts decrease carcinoma cells Y. Shen et al.1868
... Mukherjee [1] and Kazuma [22] reported that the phenolics compounds found in the flowers of C. ternatea are mainly ternatin anthocyanins and various flavanol glycosides of kaempferol, rutin, quercetin, and myricetin, which are isolated in a hydrophilic extract. Meanwhile, some fatty acids (palmitic acid, stearic acids, petroselinic acid, linoleic acid, arachidic acid, behenic acid, and phytanic acid), various phytosterols ( Figure 1) such as campestrerol, stigmasterol, β-sitosterol, and sitostanol, and tocols such as α-tocopherol and γ-tocopherol are also identified in a lipophilic extract [23]. The composition of the phytochemicals has been described in previous studies in relation to their hydrophobicity (Tables 1 and 2), although reports about the phytochemical composition quantitatively are still limited. ...
... A wide variety of polyphenols are found in the flower petals; however, the main polyphenol constituents contained within are anthocyanins [31]. The anthocyanins are blue in the petals and acylated based on delphinidin, known as ternatins isolated from C. ternatea, which are ternatin A1-A3, B1-B4, C1-C4, and D1-D3 [21,23,[32][33][34][35]. Another study reported that minor delphinidin glycosides and the preternatins A3 and C4 were isolated from the young C. ternatea [34]. ...
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Due to the beneficial health effects of polyphenolics and their limited stability during inadequate processing conditions, there is an increasing interest in their microencapsulation in order to improve the stability. As previous publications do not include a substantive review focusing on these topics, in the present work, we focused on recent reports on the topic of Clitoria ternatea flower bioactive components and the conditions under which they are microencapsulated for subsequent use in food and nutraceuticals. Our findings highlighted the importance of optimizing the variables of the microencapsulation process for optimal application.
... Penelitian lain oleh Nair et al menunjukkan bahwa ekstrak bunga telang memiliki aktifitas antiinflamasi pada cell line makrofag RAW 264.7 (Nair et al. 2015). Menurut Shen et al kandungan bioaktif bunga segar Bunga Telang yang bersifat lipofilik (kelompok fitosterol dan asam lemak )lebih banyak dibanding kandungan hidrofilik (antosianin dan flavonol glikosida) yaitu sebesar 27,67 dan 11,08 mg/100 g bunga segar (Shen et al., 2016). Kandungan senyawa fenol dalam ekstrak kering Bunga Telang menurut penelitian Singh et al antara antara 53-460 mg ekuivalen asam galat (Singh et al., 2018). ...
... Komponen bioaktif pada bunga telang yang lain yang diperkirakan memiliki manfaat fungsional adalah fenol. Kandungan utamanya adalah fitosterol, asam lemak, antosianin dan flavonol glikosida (Shen et al., 2016). Kandungan total fenol antara 53-460 mg ekivalen asam galat/ gram ekstrak kering (Adisakwattana et al., 2012;Chayaratanasin et al., 2015;Singh et al., 2018). ...
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Inhibition of atherogenesis can be done through several mechanisms, one of which is inhibiting intestinal cholesterol absorption through inhibition of NPC1L1. The activity of Telang flower extract in overcoming various problems of degenerative diseases, especially cardiovascular disease, has been proven by many scientific studies. Likewise, related to the activity of Rosella flowers. Previous research conducted by researchers showed the potential of Rosella flower extract as an LXR agonist, one of the pathways that play a role in NPC1L1 inhibition. The combination of the two extracts is expected to increase its activity as a candidate for NPC1L1 inhibitor. The purpose of this study was based on the results of the determination of total phenolic levels and the characterization of the active compounds with LC MS/MS extracts of telang flower and rosella, which are expected to be developed into further research in vivo to strengthen scientific evidence so that they can be developed into phytopharmaca preparations. Instrumentation analysis method with LC-MS/MS to identify the content of active compounds and determination of total phenolic content with Folin ciocalteu in order to obtain standardized extracts of telang flower and roselle. The results of this study showed that the total phenolic content based on the largest order was purple rosella flower extract (114.129.35 ± 266.20 g GAE), a combination of purple rosella and telang flower extract (101.760.58 ± 116.09 g GAE), telang flower extract (90.477.41 ± 107.38 g GAE) and extract red rosella flowers (84,601.00 ± 266.91 g GAE). The combination of extracts showed reduced total phenolic content compared to the single extract. The phenolic content in each extract affected the total phenolic in the combination of extracts. 8 anthocinin compounds were identified in both the telang flower extract and the rosella flower extract. Keywords: NPC1L1 inhibitor, telang, extract, Roselle, total phenolics, LC-MS/MS, atherogenesis
... Clitoria ternatea (Butterfly pea) is a member of family Fabaceae. Approximately 60 Clitoria ternatea species are found within the tropical belt, while a few species are distributed in temperate areas 7 . It has most likely initiated from tropical Asia and later spread widely in South and Central America, China, and India, where it has become naturalized. ...
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To formulate novel topical herbal gel prepared by Clitoria ternatea flowers and evaluate their antimicrobial activity. Dried, powdered flowers were extracted with ethanol using maceration method for 24 hrs. Drug-excipients compatibility study were performed for the selection of formulation excipients. Topical formulations like gel containing C. ternatea extract were formulated using combination of gelling agents such as HPMC K100M and Carbopol 934 P. Prepared gels were subjected to various evaluation parameters such as physicochemical parameters, pH, viscosity, spreadability, homogeneity, drug content uniformity, in-vitro drug diffusion, permeability and antimicrobial activity. A topical gel was successfully formulated containing bioactive ethanolic extract of C. ternatea flowers. The gel was found to be very effective as antimicrobial formulations. Prepared topical gel of Clitoria ternatea was shown pH range 6.2 to 6.6, viscosity range 480 to 603cp, spreadability range 20.62 to and homogeneous. Prepared gel was shown uniformity in drug content which ranges from 94.32 to 96.62%, zone of inhibition ranges 12mm for Bacillus spp. Prepared gel formulations were shown drug release, not more than 85% in 6 hr. As the concentration of Carbopol 934 P and HPMC K100M was increased from 0.5 to 2.0gm and 5.0 to 6.5gm respectively in the gel, antibacterial activity was synergistically improved against Bacillus spp.
... They also discovered a unique property of the mauve C. ternatea flowers, which was the accumulation of delphinidins lacking the 3 and 5 (polyacrylated) glucosyl group. Meanwhile, the research by Shen et al. [9], which was carried out only for the blue flower variety, speaks about the identification of two delphinidin derivatives and cyanidin-3-sophoroside, which they concluded was responsible for the blue color of the petals. They also found that kaempferol 3-neohesperidoside, quercetin 3-(2Grhamnosylrutinoside), and rutin were the majorly present flavonol glycoside compounds in the petals. ...
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Clitoria ternatea is a revered flower and plant in botanical science. While its health benefits are only recently gaining popularity, the plant itself has been the recipient of many traditional and indigenous medicines, including that of Ayurvedic medicine in South Asia. The peculiar property of this flower is its ability to change color depending on its pH. This review article encompasses the literature surrounding this plant and its valuable flower and attempts to cover all aspects of its benefits in the food matrix, including its existing applications. It also aims to look at the flower from a holistic perspective and imagine it as a source of future food.
... The mechanism of cisplatin resistance occurs through changes in cellular uptake, drug efflux, inhibition of apoptosis, and increased DNA repair. The resistance of cancer cells and the side effects of cisplatin are caused by its use at high doses to produce a more effective treatment [3]. Due to the rapid resistance of cancer cells to existing drugs, the search for new chemotherapeutic agents continues to this day. ...
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Clitoria ternatea L. is one of the plants whose all parts have functional benefits for the human body. The flower petals are reported to be useful as anticancer. This study aims to determine the activity of inhibiting cell proliferation, and apoptotic induction activity and identify the chemical content of isolates from Clitoria ternatea L. Analysis of the results of the MTT assay test ethanol extract of butterfly pea flower has an IC50 value of 17.46 g/mL, CTEE has an IC50 value of 13, 19 g/mL, isolate 1 and isolate 2 of EAFCT had IC50 values of 6.79 and 7.12 g/mL. The results obtained were categorized as potent cytotoxic. Isolation of EAFCT compounds using radial system chromatography then analyzed GC-MS EAFCT isolates containing pyridine hydrochloride (51.42 %) and pentanal (2.79 %) and identified compounds using UV-Vis spectrophotometry and FTIR. The results of the cell proliferation inhibition test used a series of concentrations of IC50, ⅟2 IC50, ⅟4 IC50 and ⅟8 IC50, the CTEE obtained a doubling time of 178.47; 160.86; 151.18; 150.09, EAFCT is 188.53; 164.37; 156.82; 154.79, isolate 1 EAFCT is 212.96; 174.64; 148.53; 145.82 and isolates 2 EAFCT were 190.62; 168.23; 150.24; 148.94. Analysis of the apoptotic potential test results can be concluded from CTEE, EAFCT, isolates 1 and 2 EAFCT have apoptotic potential. HIGHLIGHTS The Clitoria ternatea 1 and 2 ethyl acetate fractions had a cytotoxic effect on MCF-7 cells with IC50 values of 6.79 and 7.12 μg/mL and inhibited the growth rate of MCF-7 cells with doubling times of 212.96 and 190.62 hours Clitoria ternatea 1 and 2 isolates' ethyl acetate fraction induced apoptosis in MCF-7 cells, as indicated by the presence of orange fluorescent cells, indicating that the cells had died The Ethyl acetate fraction of Clitoria ternatea isolate contains compounds with anticancer potential, including Pyridine hydrochloride and Pentanal GRAPHICAL ABSTRACT
... The utilization of hydro-methanol has higher polarity than pure methanol, in which the polar phenolic compounds are more soluble in solvent with higher polarity [60]. During extraction, there is a tendency for the solvents to extract compounds according to their polarity as shown as a study on Clitoria ternatea petals [61]. The methanol as the polar solvent extracted hydrophilic substances including anthocyanins, kaempferol and quercetin, while the mixture of ethyl acetate and hexane as nonpolar solvent had extracted hydrophobic tocopherols, phytosterols and fatty acids. ...
Clitoria ternatea is traditionally used as medicine in Ayurveda and had been found to exhibit antibacterial activities due to its rich phytochemical contents. Due to the issue of resistant bacteria emergence and side effects of synthetic antibacterial agents, investigation of plant’s antibacterial potential is important. In this study, the methanolic C. ternatea leaves extracts were investigated for phytochemical content and antibacterial activity. Phytochemical content was investigated quantitatively focusing on the total phenolic content (TPC) and total flavonoid content (TFC), determined by Folin-Ciocalteau method and Aluminum-chloride method, respectively. The antibacterial potential of the plant’s extract was analyzed by disk-diffusion method of concentrations (12.5, 25, 50, 100 mg/mL), ampicillin and methanol act as positive control and negative control, respectively. The extraction yield of methanolic C. ternatea leaves extracts obtained by maceration method is 8.16%. The TPC and TFC of C. ternatea leaves extract are 0.66116 ± 0.43455 mg GAE /g and 0.31333 ± 0.057735 mg QE /g respectively. The disk-diffusion antibacterial assay showed no inhibitory activity of C. ternatea extracts against Escherichia coli. This might be attributed to the lack of potency of C. ternatea extracts at their current concentration, and the low content of TPC and TFC in the extracts. This had partially proved that concentration of the extracts used is crucial in antibacterial activities. Discrepancy of antibacterial results in C. ternatea observed between different studies might be attributed to the different methodologies. In conclusion, C. ternatea has been seen as a high potential plant in terms of antibacterial activity, but conditions during experiment poses high impact on the result of antibacterial assay. The findings from this study had provided valuable information to the field of phytochemistry and attempted to broaden the uses of medicinal plants, in which this can indirectly contribute to preservation of traditional knowledge and conservation of biodiversity.
Letak Indonesia baik secara geografis dan iklim sangat cocok untuk ditanam berbagai jenis tanaman yang dapat dimanfaatkan dalam kehidupan sehari-hari. Bunga telang dengan nama ilmiah Clitoria ternatea L. yang tergolong ke dalam famili Fabaceae merupakan salah satu jenis tanaman yang hidup dengan baik di Indonesia. Seluruh bagian tanaman ini dapat dimanfaatkan sebagai bahan pangan dan juga pada bidang kesehatan. Bunga ini sering dimanfaatkan sebagai pewarna makanan dan minuman alami, kandungan fitokimianya yang banyak memberikan manfaat dalam bidang kesehatan seperti antioksidan, antibakteri, anti inflamasi, analgesik, antiparasit, antihistamin, dan meningkatkan sistem imun. Pengumpulan informasi pada publikasi ini menggunakan metode studi pustaka baik secara online maupun offline, diutamakan publikasi yang terbaru kecuali yang berkaitan dengan taksonomi. Beberapa website yang digunakan yaitu google scholar, researchgate, academia, dan scopus. Hasil pencarian informasi menunjukkan bahwa telang memiliki banyak manfaat bagi manusia, diharapkan informasi yang tercantum di dalam ulasan singkat ini dapat menjadi wacana untuk budidaya bunga telang. Ulasan singkat ini berisi beberapa informasi penting tentang botani, fitokimia dan manfaat bunga telang yang dapat menjadi bahan acuan bagi penelitian selanjutnya.
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The present investigation was carried out to develop functional yogurt enriched with dried Butterfly Pea ( Clitoria ternatea L.) flower. Initially, physicochemical, microbiological, and sensorial attributes of yogurt prepared from different concentrations (0 to 3%) of Butterfly pea flower were studied. Yogurt supplemented with 1%, w/v butterfly pea flower showed better overall acceptability in sensorial terms and was optimized. The optimized BPF-rich yogurt showed 0.74 ± 0.3% ash content and 16.12 ± 0.02 total soluble solids which were higher than control yogurt. Rheological attributes (viscosity and syneresis), pH, and titratable acidity were similar in optimized BPF-rich yogurt and control yogurt. The DPPH inhibition activity (61.50%) and total phenolic content (87.23 mg GAE/g) and texture of optimized BPF-rich yogurt were better than control. The effect of storage period on free fatty acid, whey separation, acidity, total lactic acid bacteria count, coliform, yeast and mold count, and sensory parameters were studied. A significant increase was observed in free fatty acid value, acidity, and whey separation, while the lactic acid bacteria count was observed to be decreasing significantly (p < 0.05) in BPF-rich yogurt during 21 days of storage. Scanning Electron Microscopy analysis revealed better morphological characteristics and hydration properties in optimized BPF-rich yogurt. Ultra-Performing Liquid Chromatography analysis of BPF yogurt showed bioactive compounds such as delphinidin derivatives and cyanidin derivatives exhibiting functional attributes.
Color of a food is one of the major factors influencing its acceptance by consumers. At presently synthetic dyes are the most commonly used food colorant in food industry by providing more esthetically appearance and as a means to quality control. However, the growing concern about health and environmental due to associated toxicity with synthetic food colorants has accelerated the global efforts to replace them with safer and healthy food colorants obtained from natural resources (plants, microorganisms, and animals). Further, many of these biocolorants not only provide myriad of colors to the food but also exert biological properties, thus they can be used as nutraceuticals in foods and beverages. In order to understand the importance of nature-derived pigments as food colorants, this review provides a thorough discussion on the natural origin of food colorants. Following this, different extraction methods for isolating biocolorants from plants and microbes were also discussed. Many of these biocolorants not only provide color, but also have many health promoting properties, for this reason their physicochemical and biological properties were also reviewed. Finally, current trends on the use of biocolorants in foods, and the challenges faced by the biocolorants in their effective utilization by food industry and possible solutions to these challenges were discussed.
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Evaluation of sterol contents from three selected medicinal plant species of Fabaceae family growing in Rajasthan Desert was carried out. The roots, shoots and fruits of Clitoria ternatea, Sesbania bispinosa and Tephrosia purpurea were analysed for sterol contents. β- Sitosterol and Stigmasterol were isolated and identified. Maximum sterol contents were observed in shoots of Sesbania bispinosa (0.29 mg/g.d.w.), whereas minimum in roots of Tephrosia purpurea(0.15mg/g.d.w.
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Cancer hallmarks include evading apoptosis, limitless replicative potential, sustained angiogenesis, tissue invasion and metastasis. Cancer cells undergo metabolic reprogramming and inevitably take advantage of glycolysis to meet the increased metabolic demand: rapid energy generation and macromolecular synthesis. Resveratrol, a polyphenolic phytoalexin, is known to exhibit pleiotropic anti-cancer effects most of which are linked to metabolic reprogramming in cancer cells. This review summarizes various anti-cancer effects of resveratrol in the context of cancer hallmarks in relation to metabolic reprogramming.
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Proanthocyanidins (PAs) accumulate in the seeds, fruits and leaves of various plant species including the seed coats of pea (Pisum sativum), an important food crop. PAs have been implicated in human health, but molecular and biochemical characterization of pea PA biosynthesis has not been established to date, and detailed pea PA chemical composition has not been extensively studied. PAs were localized to the ground parenchyma and epidermal cells of pea seed coats. Chemical analyses of PAs from seeds of three pea cultivars demonstrated cultivar variation in PA composition. 'Courier' and 'Solido' PAs were primarily prodelphinidin-types, whereas the PAs from 'LAN3017' were mainly the procyanidin-type. The mean degree of polymerization of 'LAN3017' PAs was also higher than those from 'Courier' and 'Solido'. Next-generation sequencing of 'Courier' seed coat cDNA produced a seed coat-specific transcriptome. Three cDNAs encoding anthocyanidin reductase (PsANR), leucoanthocyanidin reductase (PsLAR), and dihydroflavonol reductase (PsDFR) were isolated. PsANR and PsLAR transcripts were most abundant earlier in seed coat development. This was followed by maximum PA accumulation in the seed coat. Recombinant PsANR enzyme efficiently synthesized all three cis-flavan-3-ols (gallocatechin, catechin, and afzalechin) with satisfactory kinetic properties. The synthesis rate of trans-flavan-3-ol by co-incubation of PsLAR and PsDFR was comparable to cis-flavan-3-ol synthesis rate by PsANR. Despite the competent PsLAR activity in vitro, expression of PsLAR driven by the Arabidopsis ANR promoter in wild-type and anr knock-out Arabidopsis backgrounds did not result in PA synthesis. Significant variation in seed coat PA composition was found within the pea cultivars, making pea an ideal system to explore PA biosynthesis. PsANR and PsLAR transcript profiles, PA localization, and PA accumulation patterns suggest that a pool of PA subunits are produced in specific seed coat cells early in development to be used as substrates for polymerization into PAs. Biochemically competent recombinant PsANR and PsLAR activities were consistent with the pea seed coat PA profile composed of both cis- and trans-flavan-3-ols. Since the expression of PsLAR in Arabidopsis did not alter the PA subunit profile (which is only comprised of cis-flavan-3-ols), it necessitates further investigation of in planta metabolic flux through PsLAR.
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Liver disease is one of the major causes of morbidity and mortality across the world. According to WHO estimates, about 500 million people are living with chronic hepatitis infections resulting in the death of over one million people annually. Medicinal plants serve as a vital source of potentially useful new compounds for the development of effective therapy to combat liver problems. Moreover herbal products have the advantage of better affordability and acceptability, better compatibility with the human body, and minimal side effects and is easier to store. In this review attempt has been made to summarize the scientific data published on hepatoprotective plants used in Saudi Arabian traditional medicine. The information includes medicinal uses of the plants, distribution in Saudi Arabia, ethnopharmacological profile, possible mechanism of action, chemical constituents, and toxicity data. Comprehensive scientific studies on safety and efficacy of these plants can revitalise the treatment of liver diseases.
Carcinoma is a major public health burden in all countries. Out of all carcinomas laryngeal carcinoma accounts for 25% of head and neck carcinoma and 2% of all human malignancies. In this study, we made an attempt to investigate the pro-oxidant activity of diosgenin on Hep2 cell line by adopting biochemical approaches. Diosgenin is a steroidal sapogenin with estrogenic and anti tumor properties. Cell viability was assessed via an MTT assay. Intracellular ROS generation was estimated spectrophotometrically. The effect of diosgenin on lipid peroxidation and the activities of antioxidants status were also analysed. Diosgenin inhibited Hep2 cell growth. The IC50 cytotoxic dose of diosgenin enhances lipid peroxidation status and decreases antioxidant levels in Hep2 cells, thereby decreasing cell malignancy or transformation and inhibits tumor promotion. Our data demonstrates that diosgenin is a potent inhibitor of Hep2 carcinoma cells, by growth inhibition,enhanced ROS generation and lipid peroxidation and decreased antioxidant activity in Hep2 cells which shows that diosgenin possess prooxidant properties.
The chemical composition of the lipophilic extracts of three Portuguese elderberries cultivars belonging to the Sambucus nigra L. species (‘Bastardeira’, ‘Sabugueira’ and ‘Sabugueiro’), was studied by gas chromatography–mass spectrometry. The influence of the harvesting season (2012 and 2013), cultivar and the ripening stage was evaluated. Regarding the amount of lipophilic extractives, they ranged from 0.56% to 1.84% of the dry weight. The major chemical families present in these fractions were triterpenoids and fatty acids accounting with 84.9–93.8% and 4.3–11.4% of the total amount of lipophilic components, respectively. The most abundant compounds, identified as elderberries components were ursolic and oleanolic acids, followed by smaller amounts of long chain aliphatic alcohols and sterols. During ripening, a similar profile of the studied chemical families was found for the two sampling seasons and the three cultivars, with an initial growth of their content followed by a systematic decrease until maturity, yet, a higher lipophilic content (p < 0.05) is reported for the 2013 harvest. Regarding mature elderberries, ‘Sabugueira’ and ‘Bastardeira’ showed higher contents of lipophilics, and particularly of triterpenic acids (p < 0.05), for the two sampling years. In-depth study of elderberries, lipophilic extractives can contribute to the valuation of this natural product, being in this study highlighted the profile of these bioactive compounds, as well, the parameters that affect their content. Additionally, since considerable amounts of unripe and overripe elderberries are produced on harvesting, and that our results showed considerable amounts of lipophilics for both ripening stages, this highlights that these wastes can be seen as a promising sources for the preparation of lipophilic extracts enriched in bioactive compounds.
Pyruvate kinase isoenzyme M2 (PKM2) is one of the most important control point enzyme in glycolysis pathway. Hence, its inhibitors and activators are currently considered as the potential anticancer agents. The effect of 28 polyphenolic compounds on the enzyme activity was investigated in vitro. Among these compounds, neoeriocitrin, (-)-catechin gallate, fisetin, (±)-taxifolin and (-)-epicatechin have the highest inhibition effect with IC50 value within 0.65-1.33 µM range. Myricetin and quercetin 3-β-d-glucoside exhibited the highest activation effect with 0.51 and 1.34 µM AC50 values, respectively. Twelve of the compounds showed inhibition effect within 7-38 µM range of IC50 value. Sinapinic acid and p-coumaric acid showed an activation effect with 26.2 and 22.2 µM AC50 values, respectively. The results propose that the polyphenolics may be the potential PKM2 inhibitors/activators, and they may be used as lead compounds for the synthesis of new inhibitors or activators of this enzyme.
Hydrophilic extracts from different parts including leaves, stalks, seeds, flowers and sprouts of 3 Amaranthus species (Amaranthus hypochondriacus, Amaranthus caudatus and Amaranthus cruentus) were characterized for their phytochemical profiles including the phenolics and betacyanins by UHPLC and LC-ESI-MS, and their antioxidant activities by FRAP and ORAC assays. The main betacyanins in Amaranthus samples were identified to be amaranthine and isoamaranthine. Eleven phenolic compounds (gallic acid, protocatechuic acid, chlorogenic acid, gentistic acid, 2,4-dihydroxybenzoic acid, ferulic acid, salicylic acid, rutin, ellagic acid, kaempferol-3-rutinoside and quercetin) were identified in the extracts of different parts of Amaranthus. The total phenolic content (TPC) ranged from 1.04 to 14.94 mg GAE/g DW; the total flavonoid content (TFC) ranged from 0.27 to 11.40 mg CAE/g DW; while the total betalain content (TBC) ranged from 0.07 to 20.93 mg/100 g DW. FRAP values ranged from 0.63 to 62.21 mu mol AAE/g DW and ORAC ranged from 30.67 to 451.37 mu mol TE/g DW. The leaves of Amaranthus showed the highest TPC, TFC, TBC, FRAP and ORAC values; while the seeds and stalks the lowest. There was a strong correlation between TPC, TBC, TFC and the antioxidant activity. The result suggests that all parts of the Amaranthus plant can be a good source of antioxidants. Crown Copyright
Considering the presence of β-sitosterol in different functional foods and the potential role of neural stem cells (NSCs) in treating neurodegenerative diseases, this study was designed to evaluate the effects of oral administration of β-sitosterol on proliferation and differentiation of NSCs. Adult mice were divided into three groups. For 2 weeks, the control group received vehicle and the low and high dose groups received 33 and 99 mg/kg/day of β-sitosterol, respectively. Harvested NSCs from the subventricular zone (SVZ) were plated in neurosphere culture and the size and number of resulting neurospheres, and also their in vitro differentiation, were evaluated. β-Sitosterol at 99 mg/kg/day significantly increased proliferation of NSCs as evidenced by their neurosphere forming frequency (719.2 ± 53.65) compared to the control group (457.6 ± 21.35). Mean neurosphere diameter was significantly larger in both high and low dose β-sitosterol groups compared to the control group. β-Sitosterol did not affect in vitro differentiation of NSCs.