Plant Material and Extraction and Isolation. The aerial parts of Lobelia sessilifolia Lamb. were collected in Yanji
City, Jilin Province of Peopleꢃs Republic of China, in August 2009 and authenticated by Prof. Minglu Deng of Changchun
University of Chinese Medicine. A voucher specimen (CC-09-0825) was deposited at the Herbarium of Changchun University
of Chinese Medicine. The aerial parts (3.8 kg) of Lobelia sessilifolia Lamb. were shade-dried, ground, and extracted with
refluxing 95% EtOH successively (57 L, 2 h, 3 times). The EtOH extract was evaporated in vacuo to yield a semisolid (970 g),
7
00 g of which was column chromatographed over silica gel using petroleum ether and EtOAc step gradient as eluents to give
several main fractions: A (125.7 g), B (85.3 g), C (21.2 g), D (12.7 g), and E (40.4 g). Fraction B was purified individually by
repeated column cromatography over silica gel to yield 1 (12 mg), 2 (442 mg), and 3 (8 mg). Fractions C and D were purified
successively with petroleum ether and EtOAc (8:2) over silica gel to afford 4 (10 mg), 5 (16 mg), and 6 (21 mg). Compounds
7
(11 mg) and 8 (15 mg) were isolated and purified from fraction E over Sephadex LH-20 with CHCl and MeOH (1:1).
3
Hydrolysis of Compound 1. Hydrolysis of the fatty acid esters of the mixtures of 1 (1 mg) was performed with 5%
KOH in MeOH (1.5 mL) under reflux for 2 h. Methyl ester derivatives of the fatty acid were prepared by the refluxing fatty
acid with 1% H SO in methanol (1.5 mL) for 1 h. The methyl ester was analyzed with GC-MS.
2
4
Gas Chromatography-Mass Spectrometry. The GC-MS analyses were carried out using an Agilent 6890N-5973
GC-MS system operating on electron impact mode (equipped with an HP 5ms capillary column 30 m ꢄꢅ0.25 mm, 0.25 mm film
thickness). He (1.0 mL/min) was used as carrier gas. The initial temperature of the column was 60ꢆC, and then it was heated
to 260ꢆC at a rate of 3ꢆC/min. The identification of the fatty acid esters was based on comparison of its EI-mass spectra with
the NIST/NBS and Wiley library spectra.
–
1
Oleanol 28-Aldehyde 3-O-ꢀ-Palmitate (1). C H O , white powder (CHCl ); mp 85–86ꢆC. IR (KBr, ꢇ , cm ):
46
78
3
3
max
+
2
1
891, 2852, 1730, 1468, 1367, 1248, 1220, 1196, 1175, 1146, 987. EI-MS m/z: 678 [M] , 422 (M – 256), 256, 239, 232, 203,
+
+
1
90, 189, 175, 135, 121, 95, 69, 57; HR-EI-MS m/z 678.7539 [M] (calcd 678.7548 [M] ). H NMR (400 MHz, CDCl , ꢁ,
3
1
3
ppm) and C NMR (125 MHz, CDCl , ꢁ, ppm), see in Table 1.
3
+
ꢀ
-Amyrin 3-O-ꢀ-Palmitate (2). White powder (CHCl ); mp 81–82ꢆC. EI-MS m/z 664 [M] [6].
3
+
Maniladiol 3-O-ꢀ-Palmitate (3). White powder (CHCl ); mp 88–89ꢆC. EI-MS m/z 680 [M] [8].
Stigmasterol (4). White crystalline needles (MeOH); mp 178–180ꢆC. EI-MS m/z 412 [M] [9].
Limettin (5). White crystalline needles (CHCl ); mp 147–148ꢆC. FAB-MS m/z 206 [M] [10].
Scoparone (6). White crystalline needles (CHCl ); mp 144–145ꢆC. EI-MS m/z 206 [M] [11].
Oleanolic Acid (7). Colorless crystalline needles (CHCl ); mp 291–292ꢆC. EI-MS m/z 456 [M] [12].
Ursolic Acid (8). Colorless crystalline needles (CHCl ); mp 286–288ꢆC. EI-MS m/z 456 [M] [13].
3
+
+
3
+
3
+
3
+
3
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2
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1
1
1
0.
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