Six steroidal saponins and two pregnane glycosides was isolated from Smilax microphylla
21
methyl-benzylamine (5 mg) and NaBH3CN (8 mg) in EtOH (1 mL)
were added. After being stirred at 40 ꢀC for 4 h followed by addition
of glacial HOAc (0.2 mL) and evaporated to dryness, the reaction
mixture was acetylated with Ac2O (0.3 mL) in pyridine (0.3 mL) for
24 h. After evaporation, 1 mL H2O was added to the residue, and
the solution was passed through an SPE cartridge washed with
H2O, H2O/MeCN (4:1, 1:1, v/v each 5 mL), successively. The H2O/
MeCN (1:1) eluate was analyzed, and the acetate derivatives were
identified by HPLC analysis with the derivative of a standard
sugar.[3,26] HPLC conditions: Inertsil ODS-3, 4.6 ꢁ 250 mm; solvent,
MeCN /H2O (3:3, v/v); flow rate: 1.0 mL/min; detection, UV absor-
bance at 230 nm. The derivatives of D-glucose and L-rhamnose
were detected at tR of 15.23 and 24.44 min, respectively.
18
O
20
19
1
4
OH
O
O
O
HO
HO
6
1'
H3C
O
1''
O
HO
O
H3C
HO
O
1''''
Acknowledgements
HO
OH
H3C
The authors acknowledge the financial support of the National
Natural Science Foundation of China (Grant Nos. 30960494 and
81160507) and Jiangxi Province NSFC (Grant No. 2009 GZY0099).
O
1'''
HO
HO
OH
Figure 3. Key HMBC correlations (H ! C) of compound 7.
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purified by Sephadex LH-20 column, and eluted with MeOH
to afford 7 (10.2 mg) and 8 (50 mg). Fr.C.4 was subjected to
Sephadex LH-20 column, eluted with MeOH, and then further
purified by C18 reverse-phase chromatographic column with
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23-oxo-pseudoprotodioscin, (25R) 26-O-b-D-glucopyranosyl-5,20-
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1
nmax KBr cm–1: 3422, 1710, 1041; H and 13C NMR spectral data
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pregna-5,16-diene-3b-ol-20-one 3-O-a-L-rhamnopyranosyl-(1 ! 2)-
[a-L-rhamnopyranosyl-(1 ! 4)]-a-L-rhamnopyranosyl-(1 ! 2)-b-D-
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1
Prosapogenin A (3),[23] white amorphous powder; H and 13C
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Protodioscin (4),[24] white amorphous powder; 1H and 13C NMR
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Methylprotodioscin (5),[9] white amorphous powder; 1H and
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1
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Acid hydrolysis of 1 and 7
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Compounds 1 and 7 (2 mg, each) in 2 M CF3COOH (5 mL) were
hydrolyzed on a water bath for 4 h. After extraction with CHCl3
(3ꢁ 5 mL), the aqueous layer was evaporated to dryness with
MeOH. The residue was dissolved in 1 mL H2O, to which L-(_)-a-
Magn. Reson. Chem. 2012, 50, 813–817
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