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Bangladesh J Pharmacol 2018; 13: 302-308
Figure 1: 1H-NMR chemical shifts of compound 3a
Yield: 56%; white solid; m.p.: 69-71°C; Rf: 0.32 (ethyl
acetate : n-hexane 2:1); IR (KBr) νmax (cm-1): 3140 (C-H
aromatic), 2931 (C-H aliphatic), 1690 (C=O ester), 1604
(C=C aromatic), 1274 (C-O ester), 1185 (C-N amine),
1099 (C-O ether) cm-1; 1H-NMR (CDCl3, 300 MHz, 25 °C,
TMS) δ (ppm): 3.02 (18H, s, -C11H3, -C12H3, -C11'H3, -
C12'H3, -C11"H3, -C12"H3), 3.77(1H, m, C1H2-O-), 4.12 (1H,
m, C1H2-O-), 4.36 (2H, d, J = 5.1 Hz, C3H2-O-), 4.42 (1H,
m, C2H-O-), 6.67 (6H, m, C7H, C9H, C7'H, C9'H, C7"H,
C9"H), 7.91 (6H, m, C6H, C10H, C6'H, C10'H, C6"H, C10"H),;
13C NMR: (75 MHz, DMSO-d6): 165.7, 154.7, 130.9,
119.8.9, 112.0, 67.8, 62.9,40.8.; EIMS: m/z (rel. abund. %),
533.25 (M+,27),; Anal. Calcd for C30H35N3O6: C, 67.52; H,
6.61; N, 7.87; O, 17.99; Found: C, 67.55; H, 6.59; N, 7.88;
O, 18.00.
(4H, d, J = 8.6 Hz, C1H2-O-, C3H2-O-), 4.44 (1H, m, C2H-
O-), 6.63 (6H, d, J = 9 Hz, C8H, C10H, C8'H, C10'H, C8"H,
C10"H), 7.91 (6H, d, J = 9 Hz, C7H, C11H, C7'H, C11'H, C7"H,
C11"H),; 13C NMR: (75 MHz, DMSO-d6): 172.4, 154.3,
131.5, 127.9, 115.3, 68.6, 63.2, 39.8, 41.8; EIMS: m/z (rel.
abund. %), 536.59 (M+,27),; Anal. Calcd for C30H32O9: C,
67.15; H, 6.01; O, 26.84; Found: C, 67.16; H, 6.00; O,
26.86.
In vitro urease assay and antimicrobial activity
A mixture of 25 μL of enzyme (jack bean urease) and
100 mm urea containing 55 μL of buffer was incubated
with 5 μL of compound (3a-3d) which concentration
was 1 mM. Incubation process was carried out at 30°C
for 15 min in 96-well plates. Indophenol method is
described in the literature (Weatherburn, 1967), in
which the activity was measured through the NH3
production. In brief, 45 μL each of 1% w/v phenol and
0.005% w/v sodium nitroprusside (phenol reagent) and
70 μL of 0.5% w/v NaOH and 0.1 % active chloride
NaOCl (alkali reagent) were added to each well. The
increasing absorbance at 630 nm was measured after 50
min, using a microplate reader (molecular device,
USA). All the assays were performed at pH 8.2 (0.01 m
Propane-1,2,3-triyltris(4-chlorobenzoate) (3c)
Yield: 75%; white solid; m.p.: 83-85°C; Rf: 0.37 (ethyl
acetate : n-hexane 2:1); IR (KBr) νmax (cm-1): 3156 (C-H
aromatic), 2981 (C-H aliphatic), 1691 (C=O ester), 1599
(C=C aromatic), 1275 (C-O ester), 1079 (C-O ether) cm-1;
1H-NMR (CDCl3, 300 MHz, 25 °C, TMS) δ (ppm): 3.92
(4H, d, J = 8.6 Hz, C1H2-O-, C3H2-O-), 4.44 (1H, m, C2H-
O-), 7.65 (6H, m, C7H, C9H, C7'H, C9'H, C7"H, C9"H), 7.99
(6H, d, J = 8.2 Hz, C6H, C10H, C6'H, C10'H, C6"H, C10"H),;
13C NMR: (75 MHz, DMSO-d6): 166.2, 139.7, 133.6,
130.4, 128.5, 127.9, 67.6, 63.5.; EIMS: m/z (rel. abund. %),
506.00 (M+,100), 508.1(M+2, 95), 510.03 (M+4, 30),; Anal.
Calcd for C24H17Cl3O6: C, 56.77; H, 3.37; Cl, 20.95; O,
18.91; Found: C, 56.76; H, 3.39; Cl, 20.94; O, 18.93.
K2HPO4.3H2O,
1 mm EDTA and 0.01 m LiCl2).
Percentage inhibition was calculated from the formula
given as follows:
100–(ODtestwell/ODcontrol) x 100
Thiourea was used as the standard inhibitor of urease.
Antimicrobial activity
Propane-1,2,3-triyltris(2-(4-methoxyphenyl)acetate)
(3d)
The agar well diffusion method was used to determine
the antibacterial activity by plant extract and its
fractions (Ahmed et al., 1998). In this procedure, 100 μL
of inoculums (diluted to 106 CFU/mL) of test culture
was mixed with 20 mL of molten sterile tryptic soya
agar. This mixture was poured into pre-sterilized petri
dishes under sterile condition. Plates were allowed to
Yield: 68%; white solid; m.p.: 5-77°C; Rf: 0.49 (ethyl
acetate : n-hexane 2:1); IR (KBr) νmax (cm-1): 2931 (C-H
aliphatic), 1691 (C=O ester), 1607 (C=C aromatic), 1275
1
(C-O ester), 1102 (C-O ether) cm-1; H-NMR (CDCl3, 300
MHz, 25 °C, TMS) δ (ppm): 3.56 (6H, s, -C5H2-, -C5'H2-, -
C5"H2-), 3.77 (9H, s, -O-C12H3, -O-C12'H3, -O-C12"H3), 3.92