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NPC
2013
Vol. 8
No. 9
1213 - 1216
Natural Product Communications
Isolation of Cycloeucalenol from Boophone disticha and Evaluation
of its Cytotoxicity
Emmanuel Adekanmi Adewusia*, Paul Steenkampb,c, Gerda Foucheb and Vanessa Steenkampa
a
Department of Pharmacology, Faculty of Health Sciences, University of Pretoria, Private Bag X323,
Arcadia 0007, South Africa
b
Natural Product Chemistry Group, Biosciences, Council for Scientific and Industrial Research,
PO Box 395, Pretoria 0001, South Africa
c
Department of Biochemistry, University of Johannesburg, Auckland Park 2006, South Africa
adewusiadekanmi@gmail.com
Received: November 21st, 2012; Accepted: June 6th, 2013
Boophone disticha (Amaryllidaceae) is widely used in traditional medicine in southern Africa. Several alkaloids, volatile oils and fatty acids have been isolated
from the plant. However, there has been no literature report of a triterpene from B. disticha. Cycloeucalenol, a cycloartane triterpene, together with its regioisomer, was isolated from the ethyl acetate extract of the bulbs using column chromatography and preparative thin layer chromatography. Structural
elucidation was carried out using 1D and 2D NMR and mass spectroscopy. The MTT and neutral red assays were used to assess the cytotoxicity of the
compound in human neuroblastoma (SH-SY5Y) cells. The compound was obtained as a mixture of two regio-isomers, which were separated for the first time
by chromatographic optimization. Integration of the 1H NMR spectrum showed that cycloeucalenol and its regio-isomer were present in a ratio of 1.04:1. A
dose-dependent decrease in cell viability was observed using both cytotoxicity assays. IC50 values of 173.0 ± 5.1 µM and 223.0 ± 6.4 µM were obtained for the
MTT and neutral red assays, respectively, indicative of the low toxicity of the compound. This work describes for the first time, the presence of triterpene
compounds from the genus Boophone.
Keywords: Amaryllidaceae, Boophone disticha, Cycloeucalenol, Cytotoxicity, SH-SY5Y cells, Regio-isomer.
Boophone disticha (L.f.) Herb, a member of the Amaryllidaceae
family, is an attractive, bulbous plant with a thick covering of dry
scales [1]. The large, round heads occur on short stems so that they
appear to grow directly from the bulb, almost at ground level. The
flowers vary from shades of pink to red and are sweetly scented [2].
The pedicels (flower stalks) elongate after flowering to form a large
seed-head. This breaks off at the top of the scape (stalk) and
tumbles across the veld dispersing the seed. The greyish green
leaves are erect, arranged in a conspicuous fan and are usually
produced after flowering [2]. B. disticha is used traditionally to treat
several diseases. Fresh scales are applied to burns and used to treat
rashes and skin disorders including eczema. It is also used to relieve
rheumatic pains, arthritic swelling, sprains, muscular strains, painful
wounds, eye conditions, headaches, anxiety, the pain of abrasions
and inflammatory conditions [3,4]. Bulb decoctions are
administered either orally or as enemas to adults suffering from
headaches, abdominal pain, weakness, sharp chest pains and
persistent bladder pains [3]. The bulb is also used in the treatment
of varicose ulcers and for the relief of urticaria, as well as a
treatment for cancer [3].
The Amaryllidaceae alkaloids, a group of isoquinoline alkaloids
are found in various Boophone species [3]. Alkaloids isolated to
date include crinine, buphanisine, buphanamine, distichamine,
buphacetine, crinamidine, lycorine, nerbowdine, undulatine,
3-O-acetylnerbowdine, buphanidrine and 6-hydroxycrinamine
[5,6]. Buphanidrine, buphanamine and distichamine have been
reported to have affinity to the serotonin transporter indicating
their potential in treatment of depression and anxiety [7,8]. Also, 6hydroxycrinamine has been shown to contain acetylcholinesterase
inhibitory activity [6]. Several other compounds have been isolated
from the plant and these include; a volatile oil containing
furfuraldehyde, acetovanillone, chelidonic acid, copper, laevulose,
18
CH3 R
19
2
3
H
HO
1
4
10
5
11
9
6
12
8
7
13 17
16
14 15
CH
29 3
H
CH3
28
Side chain (R)
30
30
21
20
21
22
23
24
26
20
22
26
25
27
27
Cycloeucalenol (1)
24
23
25
Regio-isomer of cycloeucalenol (2)
Figure 1: Structure of cycloeucalenol and its regio-isomer.
petatriacontane, ipuranol and a mixture of free and combined
fatty acids [3,9]. However, there has been no literature report of
the detection of a triterpene from B. disticha. This paper describes
the isolation and structural elucidation of a cycloartane
triterpene from B. disticha. Toxicity of the isolated compound was
determined using both the 3-[4, 5-dimethylthiazol-2-yl]-2, 5diphenyltetrazolium bromide (MTT) and neutral red uptake assays.
In addition, as the compound was obtained as a mixture of two
regio-isomers, the separation of the regio-isomers was achieved by
chromatographic optimization.
The triterpene was isolated from the ethyl acetate extracts of the
bulbs of B. disticha as white crystals. MS data showed the pseudo
1214 Natural Product Communications Vol. 8 (9) 2013
molecular ion [M + H]+ peak as the base peak at m/z 427 which
corresponds to the molecular formula, C30H50O (MW = 426.3942
Da; iFit = 0; DBE = 6). The compound was observed to be nonpolar and was dissolved in deuterated chloroform for NMR analysis
(1H, 13C and 2D experiments). The signals obtained from both the
1
H and 13C NMR spectra were complex suggesting that the isolated
compound was a mixture of two regio-isomers. Analyses of both
the NMR and MS data revealed that the structure of the isolated
compound was cycloeucalenol (1), together with its regio-isomer
(2) (Figure 1). The NMR data obtained was compared with that of
the published data on cycloeucalenol [10,11], and our extensive
literature search revealed that cycloeucalenol and its regio-isomer
have not previously been isolated from any species of Boophone.
However, this class of compound, the cycloartanes, including
cycloeucalenol, have previously been reported from Ammocharis
coranica, a member of the Amaryllidaceae family [12]. The first
literature report of a cycloartane from this family was from the plant
Crinum asiaticum var. japonicum [13].
The 1H NMR spectra of cycloeucalenol and its regio-isomer are
very similar, with the only difference observed in the position of the
double bond on the side chain. The methyl protons of the regioisomer (2) (Figure 1), Me-28 and Me-21, appeared as broad singlets
(0.95 and 0.86); Me-26 appeared as a multiplet ( H 1.64), while
Me-29 was observed as a singlet ( H 0.88). A hextet was observed
at H 2.22 (J = 7.0 Hz), while an olefinic proton, which appeared as
a doublet, was observed at H 1.00 (J = 6.6 Hz). The 1H NMR data
compares well with that of Akihisa et al. [10]. The 13C NMR
spectra of cycloeucalenol and its regio-isomer are very similar for
C-1 to C-21, with the only difference observed in the side chain
from C-22, because of the difference in position of the double bond.
C-25 is an olefinic quaternary carbon at C 150.5, while C-27 is an
exomethylene carbon at C 109.6.
Cycloeucalenol and its regio-isomer co-chromatographed. To date
there has been no literature report in which the separation of these
regio-isomers was accomplished. This study is the first to separate
these isomers into two distinct compounds, as evident from the
chromatographic profile. Integration of the 1H NMR spectrum
showed that cycloeucalenol and its regio-isomer are present in a
ratio of 1.04:1.
The continuous use and growing demand for herbal therapies have
invigorated the quest for validating the efficacy and safety or toxic
implications of medicinal plants. This is very important, as it helps
in developing safe and cheap alternative medicines. One of the
fundamental in vitro toxicological assays performed is the direct
assessment of the effects of a plant extract or compound on the
viability of a cell line. Data obtained in these assays are very useful
in selecting the most promising candidate for further development
and obtaining data for future studies [14]. The human
neuroblastoma (SH-SY5Y) cell line, which is widely used in
experimental neurological studies, analysis of neuronal
differentiation,
metabolism
and
function
related
to
neurodegenerative and neuroadaptive processes, neurotoxicity and
neuroprotection [15], was selected to assess the cytotoxicity of
cycloeucalenol and its regio-isomer. The MTT and neutral red
uptake assays were selected to determine cell viability. Both assays
were run in parallel in order to improve the reliability of the
cytotoxicity data thereby providing a more comprehensive picture
of the potential cellular toxicity through different mechanisms.
Cytotoxicity tests were carried out to assess the effect of
cycloeucalenol and its regio-isomer on the viability of the cells. A
dose-dependent effect on cell viability was observed and results
Adewusi et al.
obtained from both cytotoxicity assays were comparable (Figure 2).
IC50 values of 173.0 ± 5.1 µM and 223.0 ± 6.4 µM were obtained
for the MTT and neutral red assays, respectively. Cycloeucalenol
and its regio-isomer were observed to have high IC50 values
for both assays, which is indicative of their low toxicity.
Two cycloartane triterpenes; 25-O-acetylcimigenol-3-O- -Dglucopyranosyl (1 →2 )- -D-xylopyranoside and 25-O-acetylcimigenol-3-O- -D-galactopyranoside showed low toxicity when
tested against mouse hepatocytes, with IC50 values >100 µM [16].
This result supports the findings of the present study.
Cycloeucalenol has been reported to show anti-inflammatory,
cardiotonic and spasmolytic effects [17,18], and its low toxicity
indicates that it could be studied further as a potential lead in
developing drugs useful in treating inflammation and with
cardioprotective properties.
In conclusion, we have described the isolation of cycloeucalenol, a
cycloartane triterpene, together with its regio-isomer, from the bulbs
of Boophone disticha. The separation of both regio-isomers into two
distinct compounds is also reported for the first time. The low
toxicity of cycloeucalenol and its regio-isomer make it a suitable
agent for further testing for pharmacological activity.
Experimental
General experimental procedures: NMR spectroscopy was
performed using a 600 MHz Varian NMR spectrometer. Structural
characterizations were carried out using a combination of 1D (1H,
13
C) and various 2D experiments. The 2D experiments carried out
included DEPT, COSY, HSQC and HMBC. Chemical shifts are
reported in units of
(ppm) and coupling constants (J) are
expressed in Hz. UV-VIS detection was achieved on a WATERS
PDA scanning from 200 – 600 nm. All chemicals for UPLC-MS
were of ultra-pure LC-MS grade and purchased from Fluka
(Steinheim, Germany), while ultra-pure solvents were purchased
from Honeywell (Burdick & Jackson, Muskegon, USA). Ultra-pure
water was generated from a Millipore Elix 5 RO system and
Millipore Advantage A10 Milli-Q system (Millipore SAS,
Molsheim, France). Silica gel 60 (0.063-0.2 mm) was used for CC,
while pre-coated glass plates (Merck, SIL G-25 UV254, 20 cm x 20
cm) were used for TLC and preparative TLC. Compounds on the
TLC plates were detected under UV light at short wave (250 nm)
and long wave (365 nm) lengths, and by spraying with vanillinH2SO4 reagent. MTT and neutral red dye, purchased from Sigma
were used for the cytotoxicity assays.
Plant material: Bulbs of Boophone disticha (L.f.) Herb.
(Amaryllidaceae) were a gift from the South African National
Biodiversity Institute, Pretoria.
Extraction and isolation of cycloeucalenol and its regio-isomer:
Plant material was cut into small pieces and air-dried at room
temperature. Dried material was ground to a fine powder using an
Ika Analytical Mill (Staufen, Germany), and stored at ambient
temperature in the dark till use. Powdered plant material (250 g)
was extracted with 2.5 L of ethyl acetate for 24 h while shaking.
The extracts were filtered, concentrated using a rotary vacuum
evaporator and further dried under reduced pressure. The ethyl
acetate extract (1.4 g) was subjected to silica gel CC (65 g; particle
size 0.063 - 0.2 mm).
The separation and purification was carried out using a stepwise
gradient mixture of n-hexane: ethyl acetate starting from 100:0 until
0:100 as eluent to give 70 fractions. Fractions were collected every
Cycloeucalenol from Boophone disticha
Natural Product Communications Vol. 8 (9) 2013 1215
5 min at a rate of 1 mL/min. The fractions were pooled together
based on the similarity in their Rf values on a TLC plate to give 4
sub-fractions. Sub-fraction 2, which contained cycloeucalenol, was
further purified by CC. This sub-fraction was subjected to further
silica gel column chromatographic purification and subsequently
eluted using a stepwise gradient mixture of n-hexane: ethyl acetate,
starting from 90:10 until 0:100, to give another set of 18 fractions.
These fractions were pooled together based on the similarity in their
Rf values on a TLC plate. Cycloeucalenol and its regio-isomer (0.3
g) were obtained as white crystals. These were further analyzed
using UPLC-QTOF (mass spectrometric determination) and NMR
spectroscopy (1D and 2D experiments). The separation of the 2
regio-isomers into 2 distinct compounds was evident from the
chromatographic profile (data not shown).
Instrumental: A Waters UPLC coupled in tandem to a Waters
photodiode array (PDA) detector and a SYNAPT G1 HDMS mass
spectrometer was used to generate accurate mass data.
Chromatographic separation of the purified sample utilized a
Waters HSS C18 column (150 mm x 2.1 mm, 1.8 µm) with
temperature controlled at 60ºC. A binary solvent mixture was used
consisting of water (Eluent A) containing 10 mM formic acid
(natural pH of 2.3) and methanol (Eluent B). The initial conditions
were 40% A at a flow rate of 0.4 mL/min, which was maintained
for 1 min, followed by a linear gradient to 5% A at 12 min. The
conditions were kept constant for 3 min and then changed to the
initial conditions. The runtime was 20 min and the injection volume
was 5 µL. The PDA detector was scanned between 200 and 500 nm
(1.2 nm resolution), which collected 20 spectra per second.
The SYNAPT G1 mass spectrometer was used in V-optics and
operated in electrospray ionization mode to enable detection of
terpenes. Leucine enkephalin (50 pg/mL) was used as reference
calibrant to obtain typical mass accuracies between 1 and 3 mDa.
The mass spectrometer was operated in positive mode with a
capillary voltage of 3.0 kV, the sampling cone at 25 V and the
extraction cone at 4 V. The scan time was 0.1 sec covering the 100
to 1000 Da mass range. The source temperature was 120ºC and the
desolvation temperature was set at 400ºC. Nitrogen gas was used as
the nebulization gas at a flow rate of 800 L/h. The software used to
control the hyphenated system and for data manipulation was
MassLynx 4.1 (SCN 704).
Cells and cell culture: Human neuroblastoma (SH-SY5Y) cells
(ATCC CRL-2266) were used for the cytotoxicity studies. Cells
were cultured in Ham’s F-12 supplemented with 2% heatinactivated fetal bovine serum, penicillin (100 U/mL) and
streptomycin (100 µg/mL) at 37oC in a humidified incubator at 95%
air and 5% CO2. For use in the assay, the cells were trypsin-treated
for 10 min, decanted from culture flasks and centrifuged (200 g, 10
min). The pellet was re-suspended in 1 mL Ham’s F-12 medium
supplemented with fetal calf serum, and enumerated by staining
with trypan blue. The cells were diluted to a concentration of 1 ×
105 cells/well in Ham’s F-12 medium and 100 µL of the cell
suspension plated into each of the wells of a 96-well microtiter
plate. Ham’s F-12 medium (80 µL) was added and plates were then
incubated for 1 h at 37oC in a humidified incubator with 95% air
and 5% CO2 to allow for cellular re-attachment.
MTT assay: The MTT assay as described by Mossmann [19] was
used to measure cell viability. The principle of the assay is based on
generation of formazan (a blue product) in the mitochondria of
active cells, which is measured by photometric techniques [20]. The
compound was dissolved in 0.3%, v/v, DMSO in distilled water.
The vehicle was used as control.
Figure 2: Effect of cycloeucalenol and its regio-isomer on the viability of SH-SY5Y
cell lines as determined by the MTT and neutral red uptake assays after 72 h of
incubation.
The cells were plated into 96-well culture plates, as described
above, and treated with various concentrations of the compound
ranging from 3.125 μM to 400 μM for 72 h. Thereafter, 20 μL of
MTT solution (5 mg/mL) was added to the wells and further
incubated for 3 h. A solution (50 μL) containing 30%, w/v, N,Ndimethylformamide and 20% sodium dodecyl sulfate in water was
then added to breach the cells and dissolve the formazan crystals.
The plates were incubated overnight at 37oC, after which
absorbance was measured at 570-630 nm using a microtiter plate
reader (Labsystems Multiscan EX type 355). Wells without cells
were used as blanks and were subtracted as background from each
sample. Cytotoxicity results are expressed as the percentage cell
survival compared with the untreated control using a dose response
curve and extract concentration providing 50% inhibition (IC50) was
calculated from the graph of inhibition percentage versus extract
concentration.
Neutral red assay: The neutral red uptake assay, as described by
Borenfreund and Puerner [21], was also used to assess cell viability.
This method is based on the determination of the accumulation of
the neutral red dye in the lysosomes of viable, uninjured cells. The
compound was dissolved in 0.3%, v/v, DMSO in distilled water.
The vehicle was used as control. The cells were plated into 96-well
culture plates, as described above, and treated with various
concentrations of the compound ranging from 3.125 μM to 400 μM
for 72 h. Thereafter, 150 µL of neutral red dye (100 µg/mL)
dissolved in serum free medium (pH 6.4) was added to the culture
medium for 3 h at 37°C. Cells were washed with Phosphate
Buffered Saline (PBS), and 150 μL of elution medium
(EtOH/AcCOOH/H2O, 50%/1%/49%) was added, followed by
gentle shaking for 60 min, so that complete dissolution could be
achieved. Absorbance was recorded at 540-630 nm using a
microtiter plate reader (Labsystems Multiscan EX type 355).
Cytotoxicity results are expressed as the percentage cell survival
compared with the untreated control using a dose response curve
and extract concentration providing 50% inhibition (IC50) of cell
death was calculated from the graph.
Statistical analysis: Tests were carried out where possible at least in
triplicate and on 3 different occasions. The results are reported as
mean ± standard deviation (S.D.). Standard curves were generated
and calculation of the 50% inhibitory concentration (IC50) values
was made using GraphPad Prism Version 4.00 for Windows
1216 Natural Product Communications Vol. 8 (9) 2013
(GraphPad Software Inc.). Cytotoxicity results are expressed as the
percentage cell survival compared with the untreated control using a
dose response curve. Data obtained from mass spectroscopy were
analyzed using MassLynx 4.1 (SCN 704) software.
Adewusi et al.
Acknowledgements - We are grateful to the National Research
Foundation (NRF) of South Africa for funding the Waters UPLC
Synapt HDMS GI system as a joint venture between CSIR
Biosciences and University of Johannesburg Biochemistry
Department.
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Natural Product Communications Vol. 8 (9) 2013
Published online (www.naturalproduct.us)
In vitro Anti-diabetic Activity of Sclerocarya birrea and Ziziphus mucronata
Nuno M.H. Da Costa Mousinho, Jacob J. van Tonder and Vanessa Steenkamp
Secondary Metabolites from the Fungus Emericella nidulans
Amer H. Tarawneh, Francisco Leόn, Mohamed M. Radwan, Luiz H. Rosa and Stephen J. Cutler
A New Glucuronolactone Glycoside Phoenixoside B from the Seeds of Phoenix dactylifera
Sumbul Azmat, Rehana Ifzal, Faryal Vali Mohammad, Viqar Uddin Ahmad and Aqib Zahoor
Cancer-Suppressive Potential of Extracts of Endemic Plant Helichrysum zivojinii: Effects on Cell Migration,
Invasion and Angiogenesis
Ivana Z. Matić, Ivana Aljančić, Vlatka Vajs, Milka Jadranin, Nevenka Gligorijević, Slobodan Milosavljević and Zorica D. Juranić
Analysis of Volatile Components, Fatty Acids, and Phytosterols of Abies koreana growing in Poland
Anna Wajs-Bonikowska, Karol Olejnik, Radosław Bonikowski and Piotr Banaszczak
Cytotoxic Effects of Air Freshener Biocides in Lung Epithelial Cells
Jung-Taek Kwon, Mimi Lee, Gun-Baek Seo, Hyun-Mi Kim, Ilseob Shim, Doo-Hee Lee, Taksoo Kim, Jung Kwan Seo,
Pilje Kim and Kyunghee Choi
GC/GC-MS Analysis, Isolation and Identification of Bioactive Essential Oil Components from the Bhutanese Medicinal
Plant, Pleurospermum amabile
Phurpa Wangchuk, Paul A. Keller, Stephen G. Pyne, Malai Taweechotipatr and Sumalee Kamchonwongpaisan
Antibacterial Activity of the Essential Oil of Heracleum sibiricum
Dragoljub L. Miladinović, Budimir S. Ilić, Tatjana M. Mihajilov-Krstev, Dejan M. Nikolić, Olga G. Cvetković,
Marija S. Marković and Ljiljana C. Miladinović
Assessment of the Chemical Composition and in vitro Antimicrobial Potential of Extracts of the Liverwort Scapania aspera
Danka R. Bukvicki, Amit K. Tyagi, Davide G. Gottardi, Milan M. Veljic, Snezana M. Jankovic, Maria E. Guerzoni and Petar D. Marin
Essential Oils of Alpinia rafflesiana and Their Antimicrobial Activities
Shariha Jusoh, Hasnah Mohd. Sirat and Farediah Ahmad
Chemical Composition and Synergistic Antioxidant Activities of Essential Oils from Atractylodes macrocephala and
Astragalus membranaceus
Jinkui Li, Feng Li, Yan Xu, Wenjian Yang, Lili Qu, Qian Xiang, Cong Liu and Dapeng Li
Chemical Analysis and Antioxidant Activity of the Essential Oils of Three Piperaceae Species Growing in the Central
Region of Cuba
Elisa Jorge Rodríguez, Yanelis Saucedo-Hernández, Yvan Vander Heyden, Ernesto F. Simó-Alfonso, Guillermo Ramis-Ramos,
María Jesús Lerma-García, Urbano Monteagudo, Luis Bravo, Mildred Medinilla, Yuriam de Armas and José Manuel Herrero-Martínez
The Composition, Anti-mildew and Anti-wood-decay Fungal Activities of the Leaf and Fruit Oils of Juniperus formosana from Taiwan
Yu-Chang Su, Kuan-Ping Hsu, Eugene I-Chen Wang and Chen-Lung Ho
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Meeting/Report
Meeting Report: First National Meeting on Aloe, April 20-21, 2013, Isernia, Italy
New Perspectives in Aloe Research: from Basic Science to Clinical Application
Raffaele Capasso, Massimiliano Laudato and Francesca Borrelli
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Review/Account
Alkaloids of the South African Amaryllidaceae: a Review
Jerald J. Nair, Jaume Bastida, Carles Codina, Francesc Viladomat and Johannes van Staden
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Natural Product Communications
2013
Volume 8, Number 9
Contents
Original Paper
Page
Alternate Biosynthesis of Valerenadiene and Related Sesquiterpenes
Shashikumar K. Paknikar, Shahuraj H. Kadam, April L. Ehrlich and Robert B. Bates
A Facile Synthesis of (±)-Heliannuol-D
Tao Zhang, Liang-Zhu Huang, You-Qiang Li, Yimg-Meng Xu and Zhen-Ting Du
A New Bioactive Diterpene Glycoside from Molinaea retusa from the Madagascar Dry Forest
Alexander L. Eaton, Liva Harinantenaina, Peggy J. Brodie, Maria B. Cassera, Jessica D. Bowman, Martin W. Callmander,
Richard Randrianaivo, Roland Rakotondrajaona, Etienne Rakotobe, Vincent E. Rasamison and David G. I. Kingston
Nitric Oxide and Tumor Necrosis factor-alpha Inhibitory Substances from the Rhizomes of Kaempferia marginata
Kanidta Kaewkroek, Chatchai Wattanapiromsakul, Palangpon Kongsaeree and Supinya Tewtrakul
Biscembranoids from the Marine Sponge Petrosia nigricans
Nguyen Xuan Nhiem, Ngo Van Quang, Chau Van Minh, Dan Thi Thuy Hang, Hoang Le Tuan Anh, Bui Huu Tai, Pham Hai Yen,
Nguyen Thi Hoai, Do Cong Thung and Phan Van Kiem
Isolation of Cycloeucalenol from Boophone disticha and Evaluation of its Cytotoxicity
Emmanuel Adekanmi Adewusi, Paul Steenkamp, Gerda Fouche and Vanessa Steenkamp
Chemical Constituents from an Endophytic Fungus Chaetomium globosum Z1
Chun-Yan Zhang, Xiao Ji, Xuan Gui and Bao-Kang Huang
Determination of C-23 Configuration in (20R)-23-Hydroxycholestane Side Chain of Steroid Compounds by 1H and
13
C NMR Spectroscopy
Alla A. Kicha, Anatoly I. Kalinovsky, Alexander S. Antonov, Oleg S. Radchenko, Natalia V. Ivanchina, Timofey V. Malyarenko,
Alexander M. Savchenko and Valentin A. Stonik
Oxasetin from Lophiostoma sp. of the Baltic Sea: Identification, in silico Binding Mode Prediction and Antibacterial
Evaluation against Fish Pathogenic Bacteria
Muftah Ali M. Shushni, Faizul Azam and Ulrike Lindequist
Chemical Constituents from the Fruit Body of Chlorophyllum molybdites
Zushang Su, Ping Wang, Wei Yuan, and Shiyou Li
Pulchranins B and C, New Acyclic Guanidine Alkaloids from the Far-Eastern Marine Sponge Monanchora pulchra
Tatyana N. Makarieva, Ekaterina K. Ogurtsova, Yuliya V. Korolkova, Yaroslav A. Andreev, Irina V. Mosharova,
Ksenya M. Tabakmakher, Alla G. Guzii, Vladimir A. Denisenko, Pavel S. Dmitrenok, Hyi-Seung Lee, Eugene V. Grishin and
Valentin A. Stonik
Cloning and Characterization of a cDNA Encoding Calcium/Calmodulin-dependent Glutamate Decarboxylase from
Scutellaria baicalensis
Yeon Bok Kim, Md Romij Uddin, Do Yeon Kwon, Min-Ki Lee, Sun-Ju Kim, Chanhui Lee and Sang Un Park
Biflavonoids, Main Constituents from Garcinia bakeriana Leaves
Ahmed Al-Shagdari, Adonis Bello Alarcón, Osmany Cuesta-Rubio, Anna Lisa Piccinelli and Luca Rastrelli
Analysis of Flavonoids and Iridoids in Vitex negundo by HPLC-PDA and Method Validation
Somendu K. Roy, Khemraj Bairwa, Jagdeep Grover, Amit Srivastava and Sanjay M. Jachak
Chemical Constituents of the Leaves of Triumfetta semitriloba
Alejandra Barraza-Morales, Deisy Medrano-Nahuat, Sergio R. Peraza-Sánchez
Phytochemical Evaluation of Lythrum salicaria Extracts and Their Effects on Guinea-pig Ileum
Tímea Bencsik, Loránd Barthó, Viktor Sándor, Nóra Papp, Rita Benkó, Attila Felinger, Ferenc Kilár and Györgyi Horváth
New Flavonol Glycosides from the Leaves of Triantha japonica and Tofieldia nuda
Tsukasa Iwashina, Minoru N. Tamura, Yoshinori Murai and Junichi Kitajima
Cytotoxic Activity of Dihydrochalcones Isolated from Corema album Leaves against HT-29 Colon Cancer Cells
Antonio J. León-González, Miguel López-Lázaro, José L. Espartero and Carmen Martín-Cordero
Immunomodulatory Activities of α-Mangostin on Peripheral Blood Mononuclear Cells
Pimolkan Kasemwattanaroj, Primchanien Moongkarndi, Kovit Pattanapanyasat, Supachoke Mangmool, Ekkarat Rodpai,
Jutima Samer, Julaporn Konlata and Kasama Sukapirom
Antiplasmodial Quinones from the Rhizomes of Kniphofia foliosa
Martha Induli, Meron Gebru, Negera Abdissa, Hosea Akala, Ingrid Wekesa, Robert Byamukama, Matthias Heydenreich,
Sylvia Murunga, Ermias Dagne and Abiy Yenesew
Biphenyl Derivatives from Garcinia schomburgkiana and the Cytotoxicity of the Isolated Compounds
Chihiro Ito, Takuya Matsui, Eri Noda, Nijsiri Ruangrungsi and Masataka Itoigawa
Anticarcinogenic Effect and Carcinogenic Potential of the Dietary Phenolic Acid: o-Coumaric Acid
Alaattin Sen, Pelin Atmaca, Gulsum Terzioglu and Sevki Arslan
Bioproduction and Optimization of Rosmarinic Acid Production in Solenostemon scutellarioides through Media
Manipulation and Conservation of High Yielding Clone via Encapsulation
Ranabir Sahu, Saikat Dewanjee and Moumita Gangopadhyay
Continued inside backcover
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