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·1150· 精细化工 FINE CHEMICALS 第 36 卷

constituents from aqueous and non-aqueous systems and filtered. Then, the solvent was removed using a
through the hydrogen-bonding interaction, van der rotary evaporator at 65 ℃, and the extracts were
Waals force (hydrophobic interaction), complexation, obtained. The extracts of Malus toringoides (Rehd.)
size-sieving action, an electrostatic force and can Hughes leaves were dissolved with deionized water at
achieve separation based on differences in the different concentrations in the following adsorption/
adsorbent affinity toward adsorbate molecules [15-16] . desorption experiments.
Most importantly, macroporous resins generally use
water and food grade ethanol as solvents to desorb target Table 1 Physical and chemical properties of macroporous
compounds and thus cost less and are environmentally adsorption resins
friendly [17] . However, to our knowledge, no report has Surface area Average pore
2
discussed the use of only one type of macroporous resin Number Type /(m /g) diameter/nm Polarity
for effectively separating and purifying FMTs from the 1 HPD-100 650~700 85~90 Non-polar
extracts of Malus toringoides (Rehd.) Hughes leaves. 2 HPD-700 650~700 65~70 Non-polar
In this study, the purification of total flavonoids by
12 macroporous resins of different polarities was 3 HPD-5000 800~870 95~100 Non-polar
screened and the optimal static and dynamic adsorption- 4 D101 400~500 100~110 Non-polar
desorption conditions were obtained by optimizing the 5 AB-8 480~520 130~140 Weak-polar
parameters of the purification process. The potential 6 HPD-400 500~550 75~80 Semi-polar
anti-inflammatory effects of FMTs were evaluated in 7 HPD-750 650~700 85~90 Semi-polar
RAW 264.7 macrophages. 8 HPD-200L 500~550 84~89 Semi-polar

9 HPD-600 550~600 80 Polar
2. Materials and methods
10 ADS-F8 120~200 80~100 Polar
2.1 Samples, chemicals and reagents 11 ADS-7 ≥100 250~300 Polar
The leaves of Malus toringoides (Rehd.) Hughes 12 NKA-9 170~250 15.5~16.5 Polar
were collected from Qamdo, Tibet province, China.
Phlorizin standard (98.0% mass fraction) was purchased 2.4 Determination of the FMTs content
from Chengdu Must Biological Technology Co. Ltd. An Agilent 1120 series HPLC system (Agilent
(Chengdu, China). Acetonitrile (HPLC grade) was Scientific, Co. Santa Clara, California, USA) equipped
purchased from Merck Co. (Darmstadt, Germany). with an Agilent Zorbax Eclipse Plus C18 column
Distilled deionized water (ddH 2 O) was produced using (2.1 mm×150mm, 3.5 μm) was applied to analyse the
a MilliQ Ultrapure water-purification system (Millipore, concentration and purity of FMTs at 286 nm. The
Bedford, MA, USA). content of FMTs was determined by the content of
2.2 Adsorbents phloridzin, and the HPLC analytical conditions for
Macroporous resins, including HPD-100, HPD-700, FMTs were as follows: the mobile phase consisted of
HPD-5000, D101, AB-8, HPD-400, HPD-750, HPD- acetonitrile (phase B) and deionized water with 0.04 %
200L, HPD-600, ADS-F8, NKA-9, and ADS-7, were phosphoric acid (phase A). The elution program was as
purchased from Cangzhou Bonchem Co. Ltd. (Hebei, follows: 0~10 min, V(A)∶V(B)=78∶22; 10~20 min,
China). The physical properties of the resins are V(A)∶V(B)=78∶22 to V(A)∶V(B)=70∶30; 20~30 min,
presented in Table 1. During the synthesis process, V(A)∶V(B)=70∶30; 30~50 min, V(A)∶V(B)= 70∶30
monomers and the porogenic agents trapped inside the
pores, in order to remove them, all the resins were first to V(A)∶V(B)=50∶50; and 50~55 min, V(A)∶V(B)=
treated in absolute ethanol for 24 h and then washed 50∶50 to V(A)∶V(B)=78∶22; the flow rate and the
with deionized water to remove the ethanol, completely. column temperature were set as 1.0 mL/min and 30 ℃,
Afterwards, the resins were soaked in 1.0 mol/L NaOH respectively; the sample injection volume was 20 μL.
for 12 h, washed with deionized water (until the pH of 2.5 Static adsorption and desorption properties of
the filtrate was 7.0), and again soaked in 1.0 mol/L macroporous resins for the purification of
HCl for 12 h, and washed with deionized water (until FMTs
the pH of the filtrate was 7.0) in sequence. Finally, all Twelve aliquots (accurately weighed as 1 g) of each
the resins were dried at 80 ℃ in an electric blast resin were separately added to twelve 100 mL flasks.
drying oven (DHG-9010A, Shanghai YiHeng Scientific The resins were first activated overnight with 95%
Instruments Co. Ltd. Shanghai, China) to reach a ethanol (volume fraction), and then the ethanol was
constant weight. thoroughly washed with deionized water. The resins in
2.3 Preparation of Malus toringoides extracts each flask were soaked in 50 mL of aqueous extract
The dried leaves of Malus toringoides (Rehd.) Hughes solutions. The initial content of the flavonoid solutions
(100 g) were extracted by reflux with 2000 mL of water was 1.247 g/L. Then, the flasks were shaken by using
for 2.0 h twice. The extracted solutions were gathered a shaking incubator with a speed of 150 r/min at 25 ℃