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Introduction 

Plants of the Sapindaceae family are trees or shrubs 
generally found in tropical or subtropical regions. In 
Cameroon, they are used in traditional medicine for the 
treatment of several ailments such as skin diseases, 
dysentery and rheumatism (Abdou-Shoer et al., 1993). 
Previous phytochemical studies on plants of this family 
reported the presence of various classes of secondary 
metabolites including flavonoids, coumarins, ellagic 
acids, ceramides, sterols, and saponins as major consti-
tuents (Aubreville et al., 1973, Huang et al., 2008, Li et 
al., 2007, Rangkadilok et al., 2005, Soh et al., 2009). Some 
of these compounds were shown to exhibit interesting 
pharmacological properties including antiplasmodial, 

hemolytic, antibacterial and anti-oxidant activities 
(Huang et al., 2008, Mesquita et al., 2005, Voutquenne et 
al., 2005). Chythrantus klaineanus is a tree or shrub of 
about 1-5 m high, found in Central Africa (Abdou-
Shoer et al., 1993). No previous reports on the chemical 
constituents or biological properties of this species have 
been reported. As part of our ongoing search for 
bioactive metabolites from Cameroonian plants of the 
Sapindaceae family, we have investigated the MeOH 
extract from the mixture of the stem bark and the trunk 
of C. klaineanus.  

Xanthine oxidase (XO, EC 1.1.3.22) is a complex 
metalloflavoprotein which catalyzes the oxidation of 
hypoxanthine and xanthine into uric acid, plays a vital 

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Xanthine oxidase inhibitory activity of compounds from 
Chythrantus claneianus  
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Phytochemical investigation of the stem bark and the trunk of Chythrantus 
claneianus led to the isolation of six known compounds named β-sitosterol (1), 
umbelliferone (2), scopoletin (3), benjaminamide (4), β-sitosterol-3-O-β-D-
glucopyranoside (5) and panconoside B (6). All these compounds were 
isolated for the first time from this plant species. The chemical structures of 
isolates were elucidated on the basis of 1 and 2 D-NMR spectra and other 
spectroscopic techniques including UV–vis, FT-IR, HR-ESIMS and HR-
FABMS. The isolates were tested in vitro for their inhibitory properties 
towards xanthine oxidase enzyme. Compounds 2, 3 and 6 showed weak 
inhibitory activities on the enzyme with IC50 values ranging from 307 µM for 
compound 6 to 475 µM for compound 3, while the extract and compounds 1, 
4 and 5 showed extremely weak activities with inhibition percentage less than 
50%. 

$UWLFOH�,QIR 

Received:  21 January 2013 
Accepted:  28 January 2013 
Available Online:   3 February 2013  
 

DOI: 10.3329/bjp.v8i1.13485 

 
Cite this article: 
Gojayev AS, Bankeu JJK, Awantu AF, 
Nkanwen ERS, Ali MS, Lenta BN, 
Guliyev AA, Noungoué DT, Ngouela 
SA, Tsamo E. Xanthine oxidase inhibi-
tory activity of compounds from 
Chythrantus claneianus. Bangladesh J 
Pharmacol. 2013; 8: 78-83. 

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ZRUN�LQ�WKH�PDQQHU�VSHFLILHG�E\�WKH�DXWKRU�RU�OLFHQVRU�  



role in disorders such as hyperuricemia and gout. XO 
inhibitors may be useful for treatment of these diseases. 
For the treatment of hyperuricemia and gout allo-
purinol, oxypurinol, and febuxostat have been used 
widely (Higgins et al., 2011; Nguyen et al., 2005).  

 

Materials and Methods 

General: Isolation of compounds was done with normal 
phase column chromatography using silica gel (70-230 
mesh, Merck, Germany). Melting points were obtained 
on a Gallenkamp melting point apparatus. Optical 
rotations were measured with a JASCO DIP-360 polari-
meter. The structures of compounds were elucidated 
using spectral methods UV–vis, fourier transform 
infrared spectroscopy, 1D and 2D NMR, HR-ESIMS and 
HR-FABMS. Ultraviolet spectra were recorded on a 
Hitachi UV 3200 spectrophotometer. A JASCO 320-A 
spectrophotometer was used for scanning IR spectros-
copy using KBr pellets. 1D and 2D NMR spectra were 
run on a Bruker spectrometer operating at 75, 100, 125, 
150, 300, 400, 500, and 600 MHz, where chemical shifts 
(δ) were expressed in ppm with reference to the solvent 
signals. Electron impact mass spectra were obtained on 
a Varian MAT 311A mass spectrometer. Fast atom 
bombardment mass spectra were measured on a JEOL-
HX 110 mass spectrometer. Column chromatography 
was performed on silica gel 230-400 mesh (Merck). 
Fractions were monitored by thin layer chromato-
graphy using precoated aluminum-backed silica gel 60 
F 254 sheets. Spots were visualized under ultraviolet 
light (254 and 365 nm) or using ceric sulfate reagent. 

Plant material: The stem bark and the trunk of C. 
klaineanus were collected in April 2008 from Mount Kala 
in the Centre Region of the Republic of Cameroon. The 
plant was identified by Mr. Nana Victor, botanist at the 
National Herbarium of Cameroon, where a voucher 
specimen (no. HNC 1489) has been deposited. 

Extraction and isolation: 2.5 kg of air-dried and pow-
dered mixture of the stem bark and the trunk of C. 
klaineanus were extracted with MeOH (5 L) at room 
temperature for 48 hours and filtered. The extract was 
concentrated to dryness under vacuum to give 40 g of a 
dark brownish crude extract. The extract was extracted 
selectively with EtOAc. The EtOAc-soluble fraction was 
evaporated to dryness under vacuum to give a dry 
residue (13 g) which was then subjected to medium 
pressure liquid chromatography over silica gel 230-400 
mesh (Merck) eluted successively with n-hexane, 
mixtures of n-hexane/EtOAc, EtOAc, and EtOAc/
MeOH of increasing polarities. Subfractions (119), of 
500 mL each, were collected and combined according to 
their thin layer chromatography profiles on pre-coated 
silica gel 60 F254 plates developed with n-hexane/EtOAc 
mixtures to yield 4 fractions (F1-4). Fraction F1 was 

subjected to column chromatography over silica gel 
(Merck, 70-230 mesh) eluted with an n-hexane/EtOAc 
mixture (9.5: 0.5 to 7: 3). This resulted in the isolation of 
1 (21 mg) and 2 (63 mg). Fraction F2 was also subjected 
to successive column chromatography over silica gel 
(Merck, 70-230 mesh) eluted with a mixture of n-hexane 
and EtOAc (6: 4) to give 3 (33 mg) and 4 (151 mg). 
Fraction F3 was eluted with an n-hexane/EtOAc 
mixture of increasing polarity (5: 5 to 1: 9) to yield 5 
(230 mg). Fraction F4 was subjected to column 
chromatography over silica gel (Merck, 70-230 mesh) 
with a CH2Cl2/MeOH mixture (9.5: 0.5 to 8: 2) to yield 6 
(320 mg). 

Acid hydrolysis of compound 6: A solution of 10 mg of 
compound 6 in 10 mL MeOH/H2O (1 : 1) mixed with 5 
mL 2N HCl was refluxed at 100°C for 3 hours. After 
evaporation of the organic solvent under vacuum and 
threefold extraction of the aqueous phase with EtOAc, 
the aglycone was identified as 3,3′,4′-tri-O-methylellagic 
acid using NMR techniques. The neutralized aqueous 
portion resulted in the detection of glucose and 
rhamnose by comparison with standard sugar samples 
on thin layer chromatography plates using n-BuOH/
Me2CO/H2O (4 : 5 : 1) as solvents and using (NH4)
6Mo7O24Ce(SO4)2H2SO4 reagent for visualization. 

In vitro xanthine oxidase inhibitory activity assay: Xanthine 
(X-0626) and xanthine oxidase (EC 1.1.3.22) (from butter 
milk) were obtained from Sigma Aldrich, (Japan). The 
xanthine oxidase inhibitory activity of compounds was 
determined by measuring the rate of hydroxylation of 
the substrate (xanthine) with the formation of uric acid, 
which is a colorless product of the reaction which 
shows absorption at 295 nm (Lee et al., 1998). Briefly, 
the reaction mixture containing 10 μL of 1 mmol/L  of 
isolates was dissolved in dimethyl sulfoxide, 150 μL of 
phosphate buffer (0.05 mmol/L, pH 7.4), 0.003 units of 
xanthine oxidase dissolved in buffer (20 μL), and 20 μL 
of 0.1 mmol/L xanthine as substrate for enzyme. After 
addition of xanthine oxidase, the mixture was 
incubated for 10 min at room temperature and pre-read 
in the ultraviolet region at 295 nm. The substrate was 
added to reaction mixture, and subsequent continuous 
reading for 15 min at an interval of 1 min was observed 
(on Spectra MAX-340). The percentage inhibitory activi-
ty by the samples were determined against a DMSO 
blank and calculated using the following formula: 

 

The IC50 of the compounds as calculated using EZ-Fit 
windows-based software (Perrella Scientific Inc. 
Amherst, USA). To compare the inhibitory activities of 
the compounds, allopurinol was used as standard. The 
reaction for each compound was performed in 
triplicate. 

100100(%) ´-=
controlOD

compoundtestOD
Inhibition

 Bangladesh J Pharmacol 2013; 8: 78-83                                                                                    79 



Results and Discussion 

The MeOH extract of the stem bark and the trunk of C. 
klaineanus was fractionated and purified on silica gel 
and sephadex LH-20 columns, successively, to afford 
six known compounds. 

Compound 6 was isolated as yellow crystals. It reacted 
positively to the Molish test, indicating its glycoside 
nature. The molecular formula C29H32O17, implying 14 
degrees of unsaturation, was deduced from NMR and 
its HR-ESIMS which showed the pseudo-molecular ion 
peak at m/z = 675.15465 [M+Na]+ (calcd. for C29H32O17 + 
Na+, 675.15686). The ultraviolet spectrum in MeOH 
exhibited absorption bands at λmax 256, 360 nm, 
suggesting an ellagic acid derivative skeleton (El-
Toumy et al., 2003, Pakulski et al., 1996). The IR 
spectrum of 6 showed absorptions for hydroxy groups 
(3431 cmí1) and lactone functions (1751 cmí1), aromatic 
C=C groups (1600 cmí1), and glycosidic C–O bonds 
(1091 cmí1). The broad band decoupled 13C NMR of 
compound 6 displayed 29 carbon signals, which were 
sorted by DEPT and HSQC experiments into twelve 
methine groups [including two aromatic methine car-
bons at δC 107.5 (C-5′) and 111.7 (C-5), and ten oxyme-
thine carbons], one oxymethylene at δC 60.3, twelve 
quaternary carbons [including two lactone carbons at δC 
158.2 (C-7) and 158.3 (C-7′)], three methoxy groups and 
one methyl group at δC 18.0 (C-6ே)] (Table I). Its 1H 
NMR spectrum exhibited two deshielded singlets, 
characteristic of an ellagic acid moiety at δH 7.65 (1H, s, 
H-5′) and 7.84 (1H, s, H-5) (6), ten oxymethine and two 
oxymethylene protons at δH 3.18 (1H, t, J = 9.4 Hz, H-
4ே), 3.25 (1H, t, J = 8.8 Hz, H-4″), 3.37 (1H, brs, H-2ே), 
3.47 (2H, m, H-3″ and H-6″), 3.55 (1H, brt, J = 8.8 Hz, H-
5″), 3.64 (2H, m, H-2″ and H-6″), 3.72 (2H, m, H-3ே and 
H-5ே), 5.26 (1H, d, J = 1.3 Hz, H-1ே) and 5.46 (1H, d, J = 
7.5 Hz, H-1″) indicating the presence of two sugar units 
in 6 (6). The spectrum also exhibited three methoxy 
groups protons at δH 4.01 (3H, s, 4′-MeOH), 4.05 (3H, s, 
3′-MeOH) and 4.11 (3H, s, 3-MeOH) as well as six 
hydroxy protons. The two sugar units obtained from 
the acid hydrolysis were identified as glucose and 
rhamnose by comparing with standard sugar samples 
on thin layer chromatography plates using n-BuOH-
Me2CO-H2O (4: 5: 1) solvent system as well as the 

carbon and proton chemical shifts and the coupling 
constant pattern of the protons of the two sugar units 
compare to those of literature (Soh et al., 2009, El-
Toumy et al., 2003). 

The configuration of the anomeric protons [5.26 (1H, d, 
J = 1.3 Hz, H-1ே) and 5.46 (1H, d, J = 7.5 Hz, H-1″)] were 
determined based on their coupling constant to be α-
rhamnose and β-glucose, respectively (Soh et al., 2009, 
El-Toumy et al., 2003). HMBC spectrum was used to 
determine the junction between the rhamnose and 
glucose and between the glucose and the ellagic acid 
moiety (Figure 1). This spectrum was also used to 
determine the position of the different methoxy groups 
on the ellagic acid moiety. In fact, cross-peaks were 
observed in the HMBC spectrum between the anomeric 
proton of the glucose moiety at δH 5.46 (1H, d, J = 7.5 
Hz, H-1″) and the carbon C-4 (δC 151.1), and between 
the anomeric proton of the rhamnose moiety at δH 5.26 
(1H, d, J = 1.3 Hz, H-1ே) and the carbon C-2″ (δC 76.2). In 
addition, cross-peaks were observed between the three 
methoxy groups protons at δH 4.11 (3H, s, 3-MeOH), 
4.05 (3H, s, 3′-MeOH) and 4.01 (3H, s, 4′-MeOH), and 
carbons C-3 (δC 140.8), C-3′ (δC 141.1) and C-4′ (δC 154.3), 
respectively. The position of the glucose unit was also 
supported by the observation of a NOE between 
protons H-5 (δH 7.84) and H-1″ (δH 5.46) in the NOESY 
spectrum. On the basis of the above spectral data and 
by comparing of these values with those reported in 
literature, the identity of compound 6 was confirmed as 
panconoside B which was recently isolated from 
Pancovia pedicellaris, a plant of the same family (Soh et 
al., 2009). 

The structures of the five other known compounds 
were established as β-sitosterol (1) (Xiao et al., 2002), 
umbelliferone (2) (Ngadjui et al., 1991), scopoletin (3) 
(Cardona et al., 1992), benjaminamide (4) (Simo et al., 
2008) and β-sitosterol-3-O-β-D-glucopyranoside (5) 
(Nguyena et al., 2004) by comparing of their spectral 
and physical data with those published before (Figure 
2). 

β-sitosterol (compound 1): Colourless needles, mp 
139─140 °C. 13C NMR (100 MHz, CDCl3) δC 38.6 (C-1), 
30.7 (C-2), 73.8 (C-3), 39.7 (C-4), 141.9 (C-5), 122.9 (C-6), 
33.1 (C-7), 33.3 (C-8), 51.7 (C-9), 37.9 (C-10), 22.2 (C-11), 
41.1 (C-12), 43.5 (C-13), 58.2 (C-14), 25.3 (C-15), 29.4 (C-
16), 57.4 (C-17), 12.3 (C-18), 19.9 (C-19), 37.5 (C-20), 19.3 
(C-21), 35.1 (C-22), 27.1 (C-23), 47.3 (C-24), 30.3 (C-25), 
20.2 (C-26), 19.4 (C-27), 24.1 (C-28), 12.4 (C-29). EI-MS 
m/z 414 (64), 396 (40), 382 (14), 303 (23), 255 (20), 159 
(23), 81 (37), 43 (100). 

Umbelliferone, 7-hydoxycoumarin (compound 2): Yellowish 
solids, mp 231-233 °C. RMN 1H (500 MHz, DMSO-d6) δH 
6.20 (d, J = 9.5 Hz, H-3), 7.94 (d, J = 9.5 Hz, H-4), 7.53 (d, 
J = 8.8 Hz, H-5), 6.79 (dd, J = 8.5, 1.9 Hz, H-6), 6.71 (d, J 
= 1.9 Hz, H-8), 10.57 (7-OH). 13C NMR (125 MHz, 

80 Bangladesh J Pharmacol 2013; 8: 78-83 

O

OO

OCH3

O

O

OCH3

OCH3

O

O

OH

HO
HO

H
H

H

OH
OH

HO
H3C

H

1

23

4

6

7

1¢

3¢

4¢

5¢6¢

7¢
1²

2¢

6²

1¢² H

Figure 1: HMBC correlations in compound 6  



DMSO-d6) δC 161.2 (C-2), 113.1 (C-3), 144.5 (C-4), 111.2 
(C-4a), 129.7 (C-5), 111.4 (C-6), 160.4 (C-7), 102.1 (C-8), 
155.4 (C-8a). EI-MS m/z 78 (31), 105 (23), 134 (100), 162 
(87). 

Scopoletin, 7-hydroxy-6-methoxycoumarin (compound 3): 
Yellow crystals, mp 201-203 °C. RMN 1H (500 MHz, 
DMSO-d6) δH 6.22 (d, J = 9.5 Hz, H-3), 7.91 (d, J = 9.5 Hz, 
H-4), 7.21 (s, H-5), 6.78 (s, H-8), 3.82 (s, OMe-6), 10.31 (7-
OH). 13C NMR (125 MHz, DMSO-d6) δC 161.1 (C-2), 
112.1 (C-3), 144.9 (C-4), 111.0 (C-4a), 110.0 (C-5), 151.6 
(C-6), 145.7 (C-7), 103.2 (C-8), 149.9 (C-8a), 56.4 (OMe-6). 
EI-MS m/z 69 (38), 121 (24), 149 (53), 177 (64), 192 (100). 

B e n j a m i n a m i d e ,  ( 2 S , 3 S , 4 R , 1 2 E ) - 2 - { [ ( 2 R ) - 2 -
hydroxytetracosanoyl]amino}octadec-12-ene-1,3,4-triol 

(compound 4): Colorless crystals, mp 141-142 °C, [a]  
+34 (c 0.08, pyridine). IR (KBr) νmax 3200-3500 (OH), 
1633 (NH─C=O), 1H NMR (500 MHz, pyridine-d5) δH 
8.55 (d, J = 9 Hz, NH), 4.69 (dd, J = 9.9, 6.6 Hz, H-1a), 
4.49 (dd, J = 9.9, 6.6 Hz, H-1b), 5.27 (m, H-2), 4.44 
(overlapped dd, H-3), 4.18 (overlapped dd, H-4), 2.27 
(m, H-5a), 1.90 (m, H-5b and H-6), 1.22 (brs, H-7 to H-
10), 2.04 (m, H-11), 5.45 (m, H-12), 5.48 (m, H-13), 2.20 
(m, H-14), 1.22 (brs, H-15 to H-17), 0.84 (t, J = 7.2 Hz, H-
18), 4.55 (brdd, H-2′), 2.17 (m, H-3a′), 1.98 (m, H-3b′), 
1.90 (m, H-4′), 1.29 (brs, H-5′ to H-23′), 0.84 (t, J = 7.2 Hz, 
H-24′). 13C NMR (125 MHz, pyridine-d5) δC 62.3 (C-1), 
51.7 (C-2), 75.9 (C-3), 72.5 (C-4), 33.9 (C-5), 26.6 (C-6), 
29.6-33.9 (C-7 to C-10), 27.5 (C-11), 130.2 (C-12), 130.4 (C

-13), 27.9 (C-14), 22.9-33.0 (C-15 to C-17), 14,3 (C-18), 
175.7 (C-1′), 72.4 (C-2′), 35.5 (C-3′), 26.7 (C-4′), 29.3-33.0 
(C-5′ to C-22′), 22.9 (C-23′), 14.2 (C-24′). (+)HR-FABMS 
m/z 681.6280 (calcd. 681.6271 for C42H83NO5, [M+H]+). 

β-sitosterol-3-O-β-D-glucopyranoside (compound 5): 
Colorless crystals; mp 260-261 °C. 13C NMR (100 MHz, 
Pyridine-d5) δC 37.4 (C-1), 30.2 (C-2), 78.5 (C-3), 39.3 (C-
4), 140.9 (C-5), 121.8 (C-6), 32.1 (C-7), 32.0 (C-8), 50.3 (C-
9), 36.9 (C-10), 21.2 (C-11), 39.9 (C-12), 42.4 (C-13), 56.8 
(C-14), 24.4 (C-15), 28.4 (C-16), 56.2 (C-17), 11.9 (C-18), 
19.0 (C-19), 36.3 (C-20), 19.2 (C-21), 34.2 (C-22), 26.4 (C-
23), 46.0 (C-24), 29.4 (C-25), 19.9 (C-26), 19.4 (C-27), 23.3 
(C-28), 12.1 (C-29), 102.5 (C-1′), 75.2 (C-2′), 78.3 (C-3′), 
71.7 (C-4′), 78.2 (C-5′), 62.8 (C-6′). EI-MS m/z 414 (5), 396 
(63), 381 (24), 255 (77), 159 (39), 81 (99), 55 (100). 

Panconoside B [3,3′,4′-tri-O-methylellagic acid 4-O-[α-L-
rhamnopyranosyl-(1→2)]-β-D-glucopyranoside] (compound 
6): Yellow crystals, mp 257-259 °C. UV (MeOH) λmax 

(logε) 256 (3.72), 360 (2.47). [a]  í80.0°(c 0.5, MeOH). 
IR (KBr) νmax 3431 (O─H), 1751 (C=O), 1600 (C=C), 1091 
cmí1. 1H NMR (500 MHz, DMSO-d6) and 13C NMR (125 
MHz, DMSO-d6) spectroscopic data, see Table I. (+)-HR-
ESIMS m/ z 675.15465 (calcd. 675.15686 for C29H32O17 + 
Na+). EI-MS m/z 344 (100), 329 (20), 286 (17). 

In vitro xanthine oxidase inhibitory assay All the isolated 
compounds and the extract were tested in vitro for their 
inhibitory effects towards the xanthine oxidase enzyme, 

30
D

20
D

 Bangladesh J Pharmacol 2013; 8: 78-83                                                                                    81 

HO

H

H H

H

1

OHO O
2

NH

O

OH

OH

(CH2)19

HO

OH 4

O

H

H H

H

O

HO

HO
OHHO

5

O OHO

O

3

O

OO

O

O

O

O

O

O

O

O

OH

HO HO

OH

HO

HO

6

Figure 2: Structures of isolates 1-6 



which is a target in gouty arthritis and uric acid 
nephrolithiasis drug research. As depicted in Table II, 
results showed that, overall, all the pure compounds 

exhibited weaker xanthine oxidase inhibitory activity 
compared to the reference drug, allopurinol.  

We found for the first time that panconoside B (6) 
inhibited XO activity. The highest values were obtained 
with compound 6 (82%) and 3 (52%). The isolates 
showed IC50 values ranging from 307 μM (compound 6) 
to 475 μM (compound 3), whereas the IC50 value of the 
reference drug allopurinol was 13.70 μM.  

Lin et al. investigated the effect of the coumarin 
derivatives on the inhibition of xanthine oxidase (XO) 
activity, and the structure–activity relationships of these 
derivatives against xanthine oxiase activity. They repor-
ted that scopoletin (3) and umbelliferone (2) showed 
lower (IC50 values of >100 mM) XO inhibitory activity 
(Hsiu-Chen et al., 2007). 

All constituents of Conyza bonariensis (L.) including β-
sitosterol (1) were tested for their activity towards XOD 
inhibition. The in vitro enzyme assay demonstrated that 
this compound didn’t display XOD inhibitory activity 
(Kong et al., 2001). In another study 3β-hydroxy-
sitosterol from snake fruit (Salacca edulis Reinw.) was 
mentioned to be inactive (Leni et al., 2007). We reported 

82 Bangladesh J Pharmacol 2013; 8: 78-83 

Table I 

Comparative 1H and 13C NMR spectral data of 6 with those of panconoside B 

Position δC δH [m, J (Hz)] 

1 113.7   
2 141.9   
3 140.8   
4 151.1   

δC δH [m, J (Hz)] 

112.6   
141.7   
142.4   
151.6   

5 111.7 7.84 (s) 112.2 7.83 (s) 
6 112.2   114.2   
7 158.2   158.6   
1′ 113.7   113.1   
2′ 141.2   141.6   
3′ 141.1   141.3   
4′ 154.3   154.8   
5′ 107.5 7.65 (s) 108.0 7.65 (s) 
6′ 112.6   113.3   
7′ 158.3   158.8   
1″ 98.5 5.46 (d, J = 7.5) 99.0 5.46 (d, J = 7.5) 
2″ 76.2 3.64 (m) 76.7 3.63 (m) 
3″ 77.2 3.47 (m) 77.5 3.46 (m) 
4″ 69.6 3.25 (t, J = 8.8) 70.1 3.24 (t, J = 8.8) 
5″ 77.0 3.55 (brt, J = 8.8) 77.7 3.56 (brt, J = 8.8) 
6a″ 60.3 3.64 (m) 60.8 3.65 (m) 
6b″   3.47 (m)   3.48 (m) 
1‴ 100.2 5.26 (d, J = 1.3) 100.7 5.26 (d, J = 1.3) 
2‴ 70.4 3.37 (brs) 70.9 3.37 (brs) 
3‴ 70.3 3.72 (m) 70.8 3.72 (m) 
4‴ 71.8 3.18 (t, J = 9.4) 72.3 3.18 (t, J = 9.4) 
5‴ 68.5 3.72 (m) 69.0 3.70 (m) 
6‴ 18.0 1.11 (d, J = 6.3) 18.5 1.11 (d, J = 6.3) 
3-OMe 61.3 4.11 (s) 62.2 4.11 (s) 
3′-OMe 61.7 4.05 (s) 61.8 4.05 (s) 
4′-OMe 56.8 4.01 (s) 57.2 4.01 (s) 

Table II 

In vitro inhibition of xanthine oxidase by natural 
compounds from Chythrantus claneianus  

  Sample % Inhibi-
tion 

IC50 ± SEM 
(µM)a 

1 β-sitosterol - ND 

2 Umbelliferone 47 >500 

3 Scopoletin 52 475 ± 3.7 

4 Benjaminamide 11 ND 

5 β-sitosterol-3-O-β-D-
glucopyranoside 

- ND 

6 Panconoside B 82 307±2.3 

  MeOH Extract 43 ND 

  Allopurinolb 98.8 13.7 ± 0.2 
aIC50: Concentrations that inhibited 50% of enzymes relative to nega-
tive control; ND: Not determined; SEM: standard mean error. 
bStandard used for the assay  



previously that β-sitosterol (1), β-sitosterol-3-O-β-D-
glucopyranoside (5) and benjaminamide (4) were 
inactive (Gojayev et al., 2011). 

It is interesting to note that the active compounds were 
coumarins and their derivatives. Our results can justify 
the use of C. klaineanus by Cameroonian traditional 
healers in the treatment of inflammatory diseases such 
as rheumatism. 

 

Acknowledgments 

The authors wish to thank the Third World Academy of 
Sciences (TWAS) and the International Center for Chemical 
and Biological Studies (ICCBS). Baku State University is also 
acknowledged. The interest and support of Prof. M. I . 
Choudhary is greatly acknowledged by Anar Gojayev. 

 

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$XWKRU�,QIR 
Anar Sahib Gojayev (Principal contact) 
e-mail: anar.biochemistry@gmail.com 

 Bangladesh J Pharmacol 2013; 8: 78-83                                                                                    83 


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