Journal of Natural Products
Article
chromatography was carried out with powdered silica gel (Kieselgel
60, 230−400 mesh, Merck KGaA, Dermstadt, Germany) and styrene-
divinylbenzene (Diaion HP-20, 250−800 μm particle size, Mitsubishi
Chemical Co., Ltd.). Precoated glass plates of silica gel (Kieselgel 60,
F254, Merck Co., Ltd., Japan) and RP-18 (F254S, Merck KGaA) were
used for TLC analysis. The TLC spots were visualized under UV light
at 254 nm wavelength and spraying with dilute H2SO4 followed by
heating. Repeated HPLC was carried out mainly with a JASCO model
887-PU pump, and isolates were detected by an 875-UV variable-
wavelength detector. Reversed-phase columns for preparative separa-
tions (Tosoh TSK gel ODS-80Ts, 5 μm, 6 × 60 × 2 cm; Tosoh
Chemicals Co. Ltd., Tokyo, Japan; flow rate 45 mL/min with
detection at 205 nm) and semipreparative separations (Cosmosil
Cholester, 5 μm, 2 × 25 cm, Nacalai Co. Ltd., Kyoto, Japan; YMC-
Pack R&D ODS, 5 μm, 2 × 25 cm, YMC Co. Ltd., Kyoto, Japan; flow
rate 9 mL/min with detection at 205 nm) were used.
Fetal bovine serum (FBS) was purchased from Gibco (Grand
Island, NY, USA). Eagle’s minimum essential medium (EMEM) and
Roswell Park Memorial Institute medium (RPMI-1640) were
purchased from Nissui Pharmaceutical Co., Ltd. (Tokyo, Japan).
Penicillin and streptomycin were purchased from Meiji Seika Kaisha
Ltd. (Tokyo, Japan). L-Glutamine and D- and L-glucose were purchased
from Wako Pure Chemical Industries Ltd. (Osaka, Japan). 17β-
Estradiol and dextran-coated-charcoal (DCC) were purchased from
Sigma Chemicals Co. (St. Louis, MO, USA).
Plant Material. The leaves of Termilalia citrina were collected from
the Rangamati District in the Hill Tracts Region of Bangladesh in May
2013, with prior official permission. The leaves were identified by Mr.
Sardar Nasir Uddin, Senior Scientific Officer, National Herbarium,
Mirpur, Dhaka, Bangladesh, and a voucher specimen has been archived
in this herbarium for further reference (DACB accession no. 38094).
Extraction and Isolation. The air-dried powdered leaves of the
plant (ca. 3.4 kg) were extracted four times with hot methanol (4 × 15
L) by refluxing for 3 h each to afford a viscous mass of 608 g. The
crude extract was then suspended in 2 L of water and partitioned with
EtOAc (2 L × 3). The EtOAc extract (93 g) was subjected to silica gel
column chromatograpy using a glass column and was eluted with a
chloroform−MeOH gradient solvent system (100:0, 99:1, 98:2, 95:5,
90:10, 67:33, 50:50). Individual fractions were collected and pooled by
analyzing their TLC profiles to afford 16 combined fractions.
Terminaloside C (3): yellow, amorphous powder; [α]25 +28.1 (c
D
0.2, MeOH); UV (MeOH) λmax (log ε) 219 (4.21), 281 (3.67) nm;
ECD (c 0.2 mM, MeOH) 215 (Δε +6.8), 235 (Δε +1.2), 288 (Δε
1
−0.9) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
3, respectively; HRFABMS m/z 595.2054 [M + H]+ (calcd for
C28H35O14, 595.2027).
Terminaloside D (4): pale yellow, amorphous powder; [α]25D +45.4
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 219 (4.25), 279 (3.61) nm;
ECD (c 0.2 mM, MeOH) 215 (Δε +10.4), 230 (Δε +1.7), 285 (Δε
1
−0.4) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
3, respectively; HRFABMS m/z 631.2006 [M + Na]+ (calcd for
C29H36O14Na, 631.2002).
2-Epiterminaloside D (5): colorless, amorphous powder; [α]25
D
+119.7 (c 0.2, MeOH); UV (MeOH) λmax (log ε) 216 (4.41), 277
(3.66) nm; ECD (c 0.1 mM, MeOH) 210 (Δε +7.5), 230 (Δε +6.6),
1
281 (Δε −0.6) nm; for H and 13C NMR spectroscopic data, see
Tables 1 and 3, respectively; HRFABMS m/z 631.1981 [M + Na]+
(calcd for C29H36O14Na, 631.2002).
Terminaloside E (6): pale yellow, amorphous powder; [α]25D +38.1
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 217 (4.21), 273 (3.45) nm;
ECD (c 0.2 mM, MeOH) 212 (Δε +4.2), 238 (Δε +1.0), 280 (Δε
1
+0.5) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
3, respectively; HRFABMS m/z 579.2096 [M + H]+ (calcd for
C28H35O13, 579.2078).
Terminaloside F (7): pale yellow, amorphous powder; [α]25D +40.6
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 218 (4.26), 283 (3.69) nm;
ECD (c 0.2 mM, MeOH) 210 (Δε +12.7), 235 (Δε +3.3), 280 (Δε
1
−0.4) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
3, respectively; HRFABMS m/z 647.1933 [M + Na]+ (calcd for
C29H36O15Na, 647.1951).
Terminaloside G (8): pale yellow, amorphous powder; [α]25D +42.3
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 214 (4.40), 277 (3.52) nm;
ECD (c 0.2 mM, MeOH) 215 (Δε +10.9), 230 (Δε +1.8), 285 (Δε
1
−0.5) nm; for H and 13C NMR spectroscopic data, see Tables 2 and
3, respectively; HRFABMS m/z 625.2526 [M + H]+ (calcd for
C30H41O14, 625.2496).
Terminaloside H (9): pale yellow, amorphous powder; [α]25 +7.6
D
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 215 (4.23), 281 (3.63) nm;
ECD (c 0.1 mM, MeOH) 215 (Δε +12.6), 235 (Δε −3.4), 288 (Δε
1
−0.7) nm; for H and 13C NMR spectroscopic data, see Tables 2 and
3, respectively; HRFABMS m/z 663.2281 [M + Na]+ (calcd for
From these fractions, fraction 11 [1.5 g: eluted with chloroform−
MeOH (90:10)] was subjected to preparative HPLC using MeCN−
water (25:75) as the mobile phase, followed by semipreparative HPLC
to afford 1 [13.7 mg; tR 56 min, YMC ODS column with MeCN−H2O
(20:80)], 3 [52.1 mg; tR 65 min, YMC ODS with MeCN−H2O
(20:80)], 9 [9.0 mg; tR 107 min, YMC ODS column with MeCN−
H2O (20:80)], 10 [11.1 mg; tR 135 min, YMC ODS column with
MeCN−H2O (20:80)], 13 [4.9 mg; tR 123 min, YMC ODS column
with MeCN−H2O (20:80)], 12 [19.9 mg; tR 140 min, YMC ODS
column with MeCN−H2O (20:80)], 2 [27.2 mg; tR 144 min, YMC
ODS column with MeCN−H2O (22:78)], 4 [68.7 mg; tR 160 min,
YMC ODS column with MeCN−H2O (22:78)], 6 [6.3 mg; tR 116
min, YMC ODS column with MeCN−H2O (22:78)], 11 [7.2 mg; tR
168 min, YMC ODS column with MeCN−H2O (23:77)], 5 [1.8 mg;
tR 202 min, Cosmosil Cholester column with MeCN−H2O (22:78)],
7 [5.1 mg; tR 98 min, Cosmosil Cholester column with MeCN−H2O
(23:77)], and 8 [8.3 mg; tR 144 min, Cosmosil Cholester column with
MeCN−H2O (20:80)], respectively.
C30H40O15Na, 663.2264).
Terminaloside I (10): pale yellow, amorphous powder; [α]25 +8.0
D
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 219 (4.2), 280 (3.63) nm;
ECD (c 0.2 mM, MeOH) 210 (Δε +11.5), 232 (Δε −4.0), 286 (Δε
1
+0.4) nm; for H and 13C NMR spectroscopic data, see Tables 2 and
3, respectively; HRFABMS m/z 663.2250 [M + Na]+ (calcd for
C30H40O15Na, 663.2264).
Terminaloside J (11): yellow, amorphous powder; [α]25 +51.2 (c
D
0.2, MeOH); UV (MeOH) λmax (log ε) 212 (4.6), 279 (3.77) nm;
ECD (c 0.1 mM, MeOH) 210 (Δε +14.5), 235 (Δε −5.0), 280 (Δε
1
+0.3) nm; for H and 13C NMR spectroscopic data, see Tables 2 and
3, respectively; HRFABMS m/z 677.2399 [M + Na]+ (calcd for
C31H42O15Na, 677.2421).
Terminaloside K (12): yellow, amorphous powder; [α]25 +32.3 (c
D
0.2, MeOH); UV (MeOH) λmax (log ε) 214 (4.29), 283 (3.77) nm;
ECD (c 0.2 mM, MeOH) 210 (Δε +12.0), 243 (Δε +0.1), 288 (Δε
1
−1.0) nm; for H and 13C NMR spectroscopic data, see Tables 2 and
Terminaloside A (1): pale yellow, amorphous powder; [α]25 +3.7
3, respectively; HRFABMS m/z 663.2263 [M + Na]+ (calcd for
D
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 212 (4.17), 283 (3.82) nm;
ECD (c 0.2 mM, MeOH) 208 (Δε +5.5), 235 (Δε +1.0), 285 (Δε
C30H40O15Na, 663.2264).
6-Epiterminaloside K (13): pale yellow, amorphous powder; [α]25
D
1
−0.6) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
+52.3 (c 0.2, MeOH); UV (MeOH) λmax (log ε) 214 (4.24), 283
(3.76) nm; ECD (c 0.2 mM, MeOH) 205 (Δε +11.6), 230 (Δε +5.9),
3, respectively; HRFABMS m/z 587.1763 [M + Na]+ (calcd for
C27H32O13Na, 587.1741).
290 (Δε +0.5) nm; for H and 13C NMR spectroscopic data, see
1
Terminaloside B (2): pale yellow, amorphous powder; [α]25D +38.3
(c 0.2, MeOH); UV (MeOH) λmax (log ε) 226 (4.08), 282 (3.84) nm;
ECD (c 0.2 mM, MeOH) 210 (Δε +10.9), 230 (Δε +2.1), 285 (Δε
Tables 2 and 3, respectively; HRFABMS m/z 663.2238 [M + Na]+
(calcd for C30H40O15Na, 663.2264).
Acid Hydrolysis of Compounds 1−13 and Sugar Identi-
fication. The absolute configurations of the sugar moieties of
compounds 1−13 were identified according to the procedure
described in a previous report, with a slight modification.30 All of
1
−0.4) nm; for H and 13C NMR spectroscopic data, see Tables 1 and
3, respectively; HRFABMS m/z 579.2067 [M + H]+ (calcd for
C28H35O13, 579.2077).
H
J. Nat. Prod. XXXX, XXX, XXX−XXX