www.ijpsonline.com
5.64 (7H, m, 7H of sugar), 2.33 (3H, s, -OAc), 2.17(6H, s, 2
x -OAc), 2.12 (3H, s, -OAc), 2.06 (3H, s, -OAc), 2.04 (3H,
s, -OAc), 2.00 (3H, s, -OAc), 1.98 (3H, s, -OAc); GC-MS
(m/z), 448 (M+), 279, 207, 167, 149, 132, 104, 83.
REFERENCES
Compound D (quercitin-3-O-α-D-glucoside, 4) was
obtained on elution with methanol-ethyl acetate (1:3), 40
mg, mp: 238°12. It responded to colour reaction with Mg/
HCl. IR (K Br, νmax in cm-1): 671, 801, 907, 1220, 1371,
1439, 1634, 2364, 2931, 3484. The compound was
acetylated with Ac2O/Py: mp: 238o12 and its 1H NMR
(CDCl3) δ, ppm: 7.89 (2H, m, J 7.0 Hz, H-2’), 7.98 (2H,
dd, J 2.0 Hz, J 7.0 Hz, H-6’), 7.26 (1H, m, H-5’), 7.09 (1
H, d, J 2.0 Hz, H-8), 6.78 (1 H, d, J 2.0 Hz, H-6), 3.92-5.60
(7H, m, 7H of sugar), 2.40 (3H, s, -OAc), 2.33(3H, s, -OAc),
2.17(3H, s, -OAc), 2.12 (3H, s, -OAc), 2.06 (3H, s, -OAc),
2.04 (3H, s, -OAc), 2.00 (3H, s, -OAc), 1.98 (3H, s, -OAc);
GC-MS (m/z) 464 (M+), 279, 207, 170, 128, 97.
3.
1.
2.
4.
5.
6.
7.
Bhattacharjee, S.K., In; Handbook of Medicinal Plants. Pointer
Publishers, Jaipur, 1998, 228.
Kirtikar, K.R. and Basu, B.D., In; Indian Medicinal Plants, Vol II,
Bishan Singh Mahendra Paul Singh, Dehradun, 1984, 830.
Agarwal, V.S., Drug Plants of India, Vol. I. Kalyani Publishers, New
Delhi, 1997, 299.
Drury, C.H., In; The Useful Plants of India, International Book
Distributors, Dehradun, 1978, 353.
Singh, R.G. and Usha Kapoor, S., J. Res. Edu. Indian Med.,
1991, 10, 35.
Anand, R., Patnaik, G.K., Kulshershta, D.K. and Dhawan, B.N,
Proceeding 24 th Indian Pharmacol. Soc. Conference,
Ahmedabad, Gujarat, India, 1994, A10.
Baskar, R., Malini, M.M., Varalakshmi, P., Bal Krishna, K., Rao, R.B.,
Meenalakshmi- Malini, M. and Bhima Rao, R, Fitoterapia, 1996, 67,
121.
Singh, S., Bani, S., Singh, G.B., Gupta, B.D., Banerjee, S.K. and Singh,
B., Fitoterapia, 1997, 68, 9.
Heilbron, I., Cook, A.H., Bunbury, H.M., Hey, D.H., Eds., In;
Dictionary of Organic Compounds, Eyre and Spottiswoode, London,
1965, 1568.
Sasaki, S., In; Handbook of proton-NMR Spectra and Data, Academic
Press, Tokyo, 1986, 2618.
Buckingham, J., Eds., In; Dictionary of Natural Products, Chapman and
Hall, London, 1994, 2570.
Buckingham, J., Eds., In; Dictionary of Natural Products, Chapman and
Hall, London, 1994, 2831.
Arya, Chitra, Daniel, M. and Arya, C., Natl. Acad. Sci. Lett.
1997, 20, 38.
m
rf o
d ns
a
lo tio
n
a
w
c
i
o bl
Kaemferol-3-O-α-D-glucoside and quercitin-3-O-α-Dd
glucoside have already been reported from this plant,
e Pu ).
while dodecanoic anhydride and methyl pentacosanoate
e
are being reported for the first time from this plant. 3-O- fr
w om
o
methyl quercitin and quercitin are also being reportedor
f kn .c
from the leaves of this plant .
le ed ow
b
ACKNOWLEDGEMENTS
la M dkn
i
aCCS H.A.U.,
y e
v
Authors are thankful to Landscape Officer,
b
Hisar, for the supply of plant material. a
is ted w.m
s w
F
o
PD te h (w
is si
h
T ofa Colchicine in Six Different Species of
Estimation
of
8.
9.
10.
11.
12.
13.
13
Accepted 23 December 2006
Revised 6 March 2006
Received 14 October 2005
Indian J. Pharm. Sci., 2006, 68 (6): 804-806
Gloriosa Grown In Vivo
P. BHARATHI, D. PHILOMINA* AND S. CHAKKARAVARTHI
School of Biotechnology and Chemical Engineering, Vellore Institute of Technology, Deemed University, Vellore-632
014, India.
Three different methods for extraction of colchicine have been studied, and it was found that extraction with
petroleum ether and chloroform was the best and most reliable method. The amount of colchicine in six different
species of Gloriosa, viz., Gloriosa superba, Gloriosa rothchildiana, Gloriosa planti, Gloriosa lutea, Gloriosa
*For correspondence
E-mail: [email protected]
806
Indian Journal of Pharmaceutical Sciences
November - December 2006
www.ijpsonline.com
casuariana and Gloriosa vuchuria, has been determined using high performance liquid chromatography. Of the
six different species of Gloriosa considered for this study, Gloriosa planti exhibited the highest level of colchicines,
followed by Gloriosa lutea, Gloriosa casuariana and Gloriosa superba. The relatively high colchicine content in
the above-mentioned species of Gloriosa, these could be recommended as substitute plants for Colchicum for the
alkaloid colchicine.
Colchicine is the drug of choice to relieve acute attacks
of gout and familial Mediterranean fever1. At present
there is renewed interest in the use of colchicine as a
possible cure for cancer-related diseases2. Colchicine
itself is too toxic for human use as an antitumour drug
and hence use has been made of its derivatives, viz.,
Demecolchicine, trimethyl colchicine acid methyl ester, 2demethyl and 3-demethylthio colchicine, which are less
toxic and have been evaluated as anti-leukaemia agents.
Data collected on 3-demethylcolchicine shows this
compound to be a broad spectrum antitumour agent of
some promise. Colchicine and its analogues have been
used clinically for the treatment of certain forms of
leukaemia and solid tumour. Owing to its potent affinity
for tubulin, colchicine is used in biological and breeding
studies to produce polyploids and in tubulin-binding
assays as a positive control3-4.
grown in vivo in nursery at the host institute. One-year­
old corms from each species were collected and sliced
into small pieces for freeze drying at -20°. After 7 d, the
freeze dried plant material was ground to fine powder
and then used for extraction of colchicine.
m
rf o
d ns
a
lo tio
n
a
w
c
i
do ubl
e P ).
e
fr w m
r
o no .co
f
k w
Among the Indian medicinal plants, the corms
of
e
l
d
o
Colchicum luteum and the seeds of Iphigenia b
contain e
n
a
alkaloid, chiefly colchicine, to the extent of about
l 0.25%M Indkthe second method (Method 2) , 12 g of plant material
iavailable
a
and 0.9% respectively . These plants are not
extracted for 6 h in a Soxhlet extractor with
y in ewas
v
b
sufficient quantities to warrant any commercial
utilization.
methanol.
The extract was diluted with distilled water and
a
m
d
Gloriosa superba is another plant containing
.. then partitioned against petroleum ether, and finally the
is tecolchicine
w
A mixture of alkaloids consisting mainlysof
colchicine
aqueous phase was extracted with chloroform. The
F
w
from dried tubers of Gloriosa superba has
been
isolated
.
chloroform extract was then evaporated and the residue
o w
D
h
Hence Gloriosa L. is known
( plant of redissolved in methanol and filtered through 0.45 µm
P toforbetethea substitute
tropics to Colchicum autumnale
alkaloid
colchicine.
filter. The filtrate thus obtained constituted the test sample.
s si
i
h a for the determination of In the third method (Method 3) , 20 g of freeze dried
Several analyticalTmethods
In the first method for extraction (Method 1)1, 0.5 g of
powdered plant material was extracted twice with 25 ml of
petroleum ether with frequent shaking for 1 h, followed
each time by filtration. The solid residues were air dried
and then extracted with 10 ml of dichloromethane at
room temperature for 30 min with frequent shaking. Then
10% solution of ammonia (0.5 ml) was added to the
mixture with vigorous shaking for 10 min; the mixture
was left undisturbed for 30 min and then filtered. The
residue was washed twice with 10 ml of dichloromethane
and then combined with the filtrate. The organic phase
was evaporated to dryness and then dissolved in 1 ml of
70% ethanol to yield the test sample.
12
5
6
7
13
colchicine in pharmaceutical preparations, in biological
fluids and in plant extracts have been described8-11. The
aim of the present study is to develop an accurate and
reliable method for the extraction and the quantification
of colchicine and to identify the species of Gloriosa with
high colchicine content. Three different methods of
extraction of colchicine have been studied and the
concentration quantified using high performance liquid
chromatography (HPLC) in six different species of
Gloriosa.
Tubers of six different species of Gloriosa, viz., Gloriosa
superba, Gloriosa rothchildiana, Gloriosa planti, Gloriosa
lutea, Gloriosa casuariana and Gloriosa vuchuria, were
November - December 2006
material was extracted using 200 ml methanol in a cold
room (10°) overnight and the homogenate was centrifuged
at 1252 × g for 5 min. The methanolic extract was
evaporated to dryness and then the residues redissolved
in 50 ml water. The aqueous extract was then centrifuged
at 7826 × g for 5 min. The supernatant was first
partitioned twice against petroleum ether and then
discarded, and then once in diethyl ether, discarding the
supernatant each time. The residue was washed five times
with equal volumes of chloroform, which was retained and
evaporated to dryness. The chloroform residue was
redissolved in 95% HPLC grade methanol and then
filtered through a 0.45 µm millipore filter to yield the test
sample.
Indian Journal of Pharmaceutical Sciences
807
www.ijpsonline .com
colchicine reported was around 0.2%14. Earlier study of
Gloriosa superba revealed that colchicine levels are the
highest during the initial growth of plant, and these levels
decline during maturation 15. Comparisons with the
previous reports are difficult due to the fact that alkaloid
content also varies with locality and season. However, the
relatively high colchicine content in Gloriosa planti,
Gloriosa lutea and Gloriosa casuariana in this study
3.00
1.700
Identification of colchicine was done by comparing the
retention time of the sample with that of the standard
obtained from Sigma, USA. A Waters HPLC system
equipped with a binary pump 1525 and porous silica with
5 µm diameter C18 4.6 × 150 mm column was used for
separation. The mobile phase consisted of acetonitrile: 3%
acetic acid (60:40), at a flow rate of 1 ml/min. The peaks
eluted were detected at 245 nm and identified with
authentic standards. The HPLC method was used to
estimate the colchicine content in the six different species
of Gloriosa. Colchicine extracted by three different
methods was eluted at 1.8 min, which was ensured in
samples by comparison with the standard containing 10
mg/ml colchicine as control.
m
rf o
d ns
The concentration levels of colchicine determined in six
a
different species of Gloriosa by three extraction methods
lo tio
are represented in Table 1, and the chromatogram for
n
a
standard and colchicine extracted by the first method is
w
c
i
o bl
depicted in figs. 1 and 2 respectively. The concentration
d
of colchicine determined by extraction with petroleum
e Pu ).
ether and dichloromethane revealed that of the six
e
species considered for study, Gloriosa planti exhibited the fr
w om
o
highest value of colchicine (0.342 mg/g), followed byor
f kn .c
Gloriosa lutea, Gloriosa casuariana and Gloriosa superba
le ed ow
(0.294, 0.246, 0.211 mg/g respectively).
b
la M dkn
Extraction of colchicine using petroleumi ether and
acompared
y to e
dichloromethane was equally effective as
v
b
a for estimation
Soxhlet extraction. The sample required
s
ed ware.m
i for extraction
and the quantity of solvents consumed
t
s wwhen
F
comparatively less for the first
two methods
o
compared with the third method
extraction,
h Nevertheless,
(wand the
PDlesstofetime.
first two methods consume
is ethersandi dichloromethane can
extraction with petroleum
h
be suggested as theTmost efficient
a and reliable method for
extraction of colchicine.
Absrobance at 245 nm
2.50
2.00
1.50
1.00
1.597
2.317
0.50
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50
5.00
5.50
6.00
Retention time (min)
Fig. 1: Typical chromatogram of standard colchicine
HPLC chromatogram for standard colchicine showing retention
time of 1.8 min
1.819
1.60
1.20
1.00
0.80
1.605
0.60
0.40
0.20
2.327
1.147
Absrorbance at 245 nm
1.40
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50
5.00
Retention time (min)
Colchicine levels in Gloriosa superba corms to the level
of around 0.9% (DM) have been reported earlier13.
Gloriosa species apparently would be a better source of
commercial colchicine than Colchicum, where the level of
Fig. 2: Typical chromatogram of colchicine content of Gloriosa
superba
HPLC chromatogram for colchicine content of Gloriosa superba
showing retention time of 1.8 min
TABLE 1: COLCHICINE CONTENT IN DIFFERENT SPECIES OF GLORIOSA
Species
Gloriosa
Gloriosa
Gloriosa
Gloriosa
Gloriosa
Gloriosa
Method 1 Colchicine (mg/g)
superba
rocthcildiana
planti
lutea
casuariana
vuchuria
0.211
0.162
0.342
0.294
0.246
0.150
±
±
±
±
±
±
0.001
0.001
0.047
0.019
0.032
0.030
Method 2 Colchicine (mg/g)
0.206
0.151
0.254
0.280
0.217
0.132
±
±
±
±
±
±
0.035
0.039
0.037
0.017
0.041
0.027
Method 3 Colchicine (mg/g)
0.137
0.136
0.141
0.166
0.142
0.145
±
±
±
±
±
±
0.029
0.017
0.016
0.014
0.034
0.017
Colchicine content in six different species of Gloriosa by three different methods of extraction
808
Indian Journal of Pharmaceutical Sciences
November - December 2006
www.ijpsonline.com
6.
encourages the cultivation of these species under suitable
conditions.
7.
ACKNOWLEDGEMENTS
8.
9.
The authors are grateful to the Management of Vellore
Institute of Technology, Vellore, for providing support to
carry out this work and the Technology Business
Incubator at this Institute for the chromatographic analysis.
10.
11.
12.
REFERENCES
13.
14.
15.
Sarin, Y.K., Jamwal, P.S., Gupta, P.K. and Atal, C.K., Curr. Sci.,
1974, 43, 87.
Clewer, H.W.V., Green, S.S. and Tuttin, F., J. Chem. Soc.
(London), 1915, 107, 835.
Sutlupinar, N., Husek, A., Potesilova, H., Dvorackova, S., Hanus, V.,
Sedmera, P. and Simanek, V., Planta Med., 1988, 54, 243.
Poulev, A., Deus-Neumann, B., Bombardelli, E. and Zenk, M., Planta
Med., 1994, 60, 77.
Ondra, P., Valka, I., Vicar, J., Sutlupinar, N. and Simanek, V., J.
Chromatogr., 1995 704, 351.
Fayyad, M., Alali, F., Alkofahi, A. and Tell, A., Nat. Prod. Lett.,
2002, 16, 395.
Hayashi, T., Yoshida, K. and Sano, K., Phytochemistry 1988, 27,
1371.
Finnie, J.F. and Van Staden, J., J. Plant Physiol., 1991, 138, 691.
Bellet, P. and Gaignault, J.C., Ann. Pharm. Fr., 1985 43, 345.
Thakur, R.S., Potesilova, H. and Santavy, F., Planta Med., 1975, 28, 201.
m
rf o
d ns
a
lo tio
n
a
w
c
i
do ubl
e P ).
e
fr w m
r
Microwave Assisted Synthesis
co Pharmacological
fo kno and
.
w and Benzofuro
le edNaphtho
Evaluation of some Potent
o
b
nOxadiazalyl
la Mand
Thiazolyl, Oxazolyl,aiThio
k
y ed
v
b
Derivatives
a
si ted w.m
s w
F
o
PD te h (w
is si
h
T a
1.
2.
3.
4.
5.
Alali, F., Tawaha, K. and Qasaymch, R.M., Phytochem. Anal.,
2004, 15, 27.
Evans, D.A., Tanis, S.P. and Hart, D.J., J. Amer. Chem. Soc.,
1981, 103, 5813.
Trease, S.E. and Evans, D., In; Pharmacognosy, 12th Edn., Balliere
Tindali, London, 1983, 593.
Poutaraud, A. and Champay, N., Rev. Suisse. Agric., 1995, 27, 93.
Kapadia, V.H., Dev, S., Rao, R.S. and Ansari, M.Y.,
Phytochemistry, 1972, 11, 1193.
Accepted 23 December 2006
Revised 6 March 2006
Received 15 October 2005
Indian J. Pharm. Sci., 2006, 68 (6): 806-809
D. B. ARUNA KUMAR, G. K. PRAKASH 1 , B. P. NANDESHWARAPPA 2 , B. M. KIRAN, B. S. SHERIGARA 2
AND K. M. MAHADEVAN*
Department of PG Studies and Research in Chemistry, School of Chemical Sciences, Kuvempu University, Jnana
Sahyadri, Shankaraghatta-577 451, 1Institute of Wood Science and Technology, Malleshwaram, Bangalore-560 003,
2
Department of PG Studies and Research in Industrial Chemistry, School of Chemical Sciences, Kuvempu University,
Jnana Sahyadri, Shankaraghatta-577 451, India.
The substituted 2-acetyl benzofurans on bromination gave substituted 2-(2-bromoacetyl) benzofurans 2a-f. These
on reaction with thiourea, urea, thiosemicarbazide and semicarbazide in presence of sodium acetate under microwave
irradiation resulted 3a-f, 4a-f, 5a-f and 6a-f respectively. The structures of newly synthesised compounds have
been established by elemental analysis, spectral data and screened for antimicrobial, anthelmintic, antiinflammatory
and analgesic activities.
Benzo[b]furan derivatives are an important class of
organic compounds, which are known to be present in
many natural products1 and posses physiological activity.
They find applications in agrochemicals 2,3 ,
*For correspondence
E-mail: [email protected]
November - December 2006
pharmaceuticals4-10 and cosmetics11,12. Benzo[b]furans are
building blocks of optical brighteners13. Many of the
natural benzo[b]furans have physiological,
pharmacological and toxic properties and as a result,
there is continuing interest in their chemical synthesis14-15.
Cyclisation reactions of various types have been used to
produce substituted benzo[b]furans16,17.
Indian Journal of Pharmaceutical Sciences
809