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Q. HU ET AL.
the compounds 17, 19, 11, 7, 20, 22, 12 and 13 showed moderate activities. The HPLC fin-
gerprint analysis of the fruits showed that they contained mostly the same active compo-
nents as those extracted from the leaves of M. hupehensis (see Supplemental material).
3. Experimental
3.1. General
UV spectra were recorded on a Shimadzu UV-2600 spectrophotometer. IR spectra were
recorded on a Bruker FT-IR Tensor-27 infrared spectrophotometer with KBr pellets. One-
dimensional (1D) and two-dimensional (2D) NMR spectra were recorded on a Bruker Avance
DRX-600 spectrometer using the residual non-deuterated solvent signal as an internal stand-
ard. ESIMS and HRESIMS were recorded on an Agilent 6520 Q-TOF. HPLC was performed on
a Shimadzu LC-6AD with a Shimadzu C18 (20 × 250 mm) column. Silica gel (100–200 and
200–300 mesh, Qingdao Marine Chemical Co., Ltd., People’s Republic of China) and MCI gel
CHP20/P120 (75–150 μm, Mitsubishi Chemical Corporation, Japan) were used for column
chromatography.
3.2. Plant material
The leaves of M. hupehensis were collected from Shansong Biological Products Co., Ltd. (Linyi,
China), in September 2014. The leaves were identified by Prof. Lan Xiang, and a voucher
specimen (MH201409) has been deposited in the Herbarium of the School of Pharmaceutical
Sciences, Shandong University.
3.3. Extraction and isolation
The aired-dried leaves of M. hupehensis (10 Kg) were extracted with EtOH/H2O (80/20 v/v,
twice, 2 h each time) under reflux conditions to result a crude extract, which was further
subjected to a HPD-100 macroporous adsorption resin chromatography eluted successively
with aqueous EtOH gradient system (0, 30 and 95%). The 30% EtOH fraction (the total
polyphenols) was separated on a polyamide column (EtOH/H2O; 0, 15, 25, 50, 100%) to result
five subfractions, fraction A1-A5. Fraction A1 was separated on a Sephadex LH-20 column
and then on semi-preparative HPLC (15% ACN : H2O) to yield compounds 10 (36 mg), 12
(9 mg), 13 (34 mg), and 22 (24 mg).The main component phlorizin (8; 300 g) was isolated
from fraction A2. Fraction A3 was subjected to silica gel columns then to Sephadex LH-20
columns to yield compounds 7 (20 mg), 16 (80 mg), 17 (16 mg), 18 (60 mg) and 20 (20 mg).
Fraction A4 was subjected to a Sephadex LH-20 column and then semi-preparative HPLC to
yield compounds 19 (10 mg) and 21 (20 mg). Fraction A5 was subjected to a Sephadex LH-20
column to yield compounds 6 (1 g) and 15 (40 mg). The 95% EtOH fraction was suspended
in H2O, respectively, partitioned with petroleum ether, EtOAc, CH2Cl2 and n-BuOH to result
four organic fractions. The CH2Cl2 soluble fraction was subjected to silica gel columns to
yield compound 14 (20 mg).The EtOAc soluble fraction was separated by silica gel columns
and RP-C18 columns to yield compounds 2 (15 mg) and 11 (25 mg). The n-BuOH soluble
fraction was separated by RP-C18 columns and semi-preparative HPLC to isolate compounds
1 (15 mg), 3 (10 mg), 4 (6 mg), 5 (8 mg) and 9 (11 mg).