D101 Macroporous Resin Column Research Articles

Comprehensive Screening and Characterization of α-glucosidase Inhibitory Components in the Edible Medicinal Plant Pholidota cantonensis Rolfe Using UPLC-Q-TOF-MS/MS Analysis and Molecular Docking.

Pholidota cantonensis Rolfe is an edible medicinal plant in the genus Pholidota of the family Orchidaceae. This plant is used to prepare medicated food in China and has been reported to possess anti-α-glucosidase activity. To date, little is known about the active substances responsible for the observed anti-α-glucosidase activity. In the present study, we aimed to screen and characterize the α-glucosidase inhibitory fraction of P. cantonensis using ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) analysis and molecular docking. As a result, the 50% ethanol fraction obtained from D101 macroporous adsorption resin column chromatography (D50 fraction) had the highest total phenol content (353.83 ± 6.06mg GAE/g) and the most prominent α-glucosidase inhibitory activity (IC50 = 30.01 ± 7.30µg/mL). Forty-five compounds were identified from the D50 fraction by using UPLC-Q-TOF-MS/MS analysis. Molecular docking results showed that six main constituents, namely, crepidatin, 2,7-dihydroxy-4-methoxyl-9,10-dihydrophenylene, 4,4',5,6-tetrahydroxystilbene, 4,7-dihydroxy-2-methoxyl-9,10-dihydrophenylene, (-)-lariciresinol, and thunalbene, in the D50 fraction occupied the catalytic sites of α-glucosidase through strong hydrophobic interactions, hydrogen bonding, and other patterns. The binding energies were between - 29.95 and - 11.41kJ/mol, indicating good binding between the tested compounds and α-glucosidase. The active ingredients responsible for the α-glucosidase inhibitory activity may include phenanthrenes, stilbenes, dibenzyls, and lignans. The D50 fraction has potential value for developing innovative drugs for the prevention and treatment of diabetes mellitus (DM) and is worthy of in-depth research.

Impact of Liparis nervosa extract on neuroinflammation mediated by LPS-induced BV-2 microglial cells and its bioactive components analysis

To investigate the impact and potential mechanisms of extracts from different parts of Liparis nervosa on neuroinflammation by lipopolysaccharide(LPS)-induced BV-2 microglial cells. The materials of L. nervosa were subjected to crushing, ethanol extraction, and concentration to obtain an alcohol extract. Subsequently, the extract was further extracted to obtain petroleum ether extract, ethyl acetate extract, N-butanol extract, and aqueous phase extract. The ethyl acetate extract was separated into distillate(1)-(6)using D101 macroporous resin column chromatography. The experiment was divided into control group, LPS model group, L. nervosa extract group, and LPS + L. nervosa group. LPS was utilized to induce a neuroinflammatory cell model in BV-2 microglial cells. The Griess test was utilized for detecting the production of nitric oxide(NO) in the cell supernatant. Cell viability was detected by MTT assay. The release of interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in the cell supernatant was quantified using ELISA.RT-qPCR was utilized to assess the m RNA levels of pro-inflammatory cytokines inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), interleukin( IL)-6, IL-1β, and TNF-α. The protein expression of i NOS, COX-2, nuclear factor kappa-B p65(p65), p-p65, extracellular signal-regulated kinase(ERK), p-ERK, c-jun N-terminal kinase(JNK), p-JNK, p38 mitogen-activated protein kinase(p38), and p-p38 MAPK(p-p38) were also evaluated by Western blot. The chemical composition of active substances in L. nervosa was analyzed using the UHPLC-Q-Exactive Orbitrap technology and literature comparison. Our findings indicate that extracts from different parts of L. nervosa exhibit a significant reduction in the release of NO from LPS-induced BV-2 microglial cells.Specifically, the ethyl acetate extract demonstrates the most notable inhibitory effect without causing cell toxicity. Additionally, the distillate(6) extracted from the ethyl acetate exhibits a reduction in the m RNA and protein levels of i NOS, COX-2, IL-6, IL-1β, and TNF-α in a dose-dependent manner, and it inhibits the protein expression of p-p65, p-ERK, p-p38, and p-JNK in LPS-induced BV-2 microglial cells. A total of 79 compounds in the distillate(6) were identified by mass spectrometry, including 12 confirmed compounds with anti-inflammatory effects. This study confirmed the remarkable efficacy of L. nervosa extract in the treatment of neuroinflammation, which may be achieved through the inhibition of NF-κB and MAPK signaling pathways.

Increasing minor ginsenosides contents and enhancing neuroprotective effects of total ginsenosides fermented by Lactiplantibacillus plantarum

Minor ginsenosides have been proven to have higher pharmacological activity than the major ginsenosides. The transformation of major ginsenosides to minor ginsenosides by lactic acid bacteria was considered to be a promising method. Therefore, this study focuses on utilizing glycosidase-producing Lactiplantibacillus plantarum GLP40 to transform total ginsenosides (TG) and increase the content of minor ginsenosides, as well as investigate the neuroprotective effects of fermented total ginsenosides (FTG). After 21d fermentation, the transformation products were purified using D101 macroporous resin column chromatography, and identified by HPLC and LC-MS analyses. The neuroprotective effect of FTG was evaluated using MPTP-induced neural injury mice model. Lact. plantarum GLP40 fermentation increased the contents of minor ginsenosides in TG, such as Rg3, Rh2, CK, and Rk3. FTG showed stronger alleviation of 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Hydrochloride (MPTP) induced memory loss and dyskinesia in mice, and inhibited tyrosine hydroxylase (TH) depletion and ionized calcium binding adapter molecule 1 (Iba-1) production than TG. Further, FTG significantly increased serum IL-10 levels and inhibited the expression of pro-inflammatory cytokines compared to TG. Moreover, FTG treatment activated the anti-apoptotic PI3K/AKT/mTOR signaling pathway and inhibited the expression of the inflammatory NF-κB/COX-2/iNOS pathway. In conclusion, Lact. plantarum GLP40 fermentation enhances the neuroprotective effects of total ginsenosides by increasing minor ginsenosides. FTG protected MPTP induced neural injury in mice by regulating inflammation and cell apoptosis signaling pathways.

Identification of the chemical components of ethanol extract of Chenopodium ambrosioides and evaluation of their in vitro antioxidant and anti tumor activities

Purpose: To determine the characteristic chemical components of the ethanol extract of Chenopodium ambrosioides and evaluate their antioxidant and anti-tumor effects in vitro.
 Methods: The plant powder (5 g) was extracted with 1 L of 80 % ethanol at room temperature for 45 min, and then placed at 60 oC at varying microwave power and duration to obtain optimal extraction conditions. Characteristic chemical components were detected using ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS). Kaempferitrin was isolated from the 80 % ethanol extract using a D101 macroporous resin column, and its content was assessed by high performance liquid chromatography (HPLC). The antioxidant effect of kaempferitrin was evaluated by its ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radicals, while its anti-proliferation activity in human liver cancer cells SMMC-7721 was determined using cell counting kit-8 (CCK-8) reagent.
 Results: Three characteristic components of ethanol extract of C. ambrosioides were obtained, namely, kaempferitrin, kaempferol-3-O-apigenin-7-O-rhamnoside and kaempferol-3-O-acetylapigenin-7-O-rhamnoside. Kaempferitrin was shown to possess strong DPPH radical and moderate ABTS radical scavenging activities. Kaempferitrin significantly inhibited the proliferation of SMMC-7721 cells at doses of 4 and 8 μg/mL, with half-maximal concentration (IC50) of 0.38 μM (p < 0.05).
 Conclusion: Kaempferitrin extracted from C. ambrosioides has antioxidant and anti-tumor activities. The results reported here indicate that C. ambrosioides may have potential use in herbal medicine practice.

Nanoparticles derived from Scutellaria barbata and Hedytois diffusa herb pair and their anti-cancer activity

Ethnopharmacological relevanceThe Scutellaria barbata D.Don and Hedyotis diffusa Willd. (SH) herb pair is extensively used in Traditional Chinese medicine for treating cancer, which has a long history in clinical empirical prescriptions, such as "kang ai ping wan (抗癌平丸)", "er ban er bai decoction (二半二白汤)". The ambiguous pharmacological ingredients and compatibility mechanism of SH herb pair promoted the exploration. Aim of the studyThe characteristics of phytochemicals self-recognition and self-assembly is used to quickly find the active nanoparticles in SH herb pair. And it provides a new strategy for the developing new drugs of TCM. MethodsThe SH-extracted nanoparticles (NPs) were gained by centrifugating from the 70% fractionated that the biological activity-guided by D101 macroporous resin column. The micromorphology of active NPs was observed with transmission electron microscopy (TEM). Dynamic light scattering (DLS), Ultraviolet−visible (UV-vis) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD) and element analysis (EA) were applied for characterizing NPs. The main components were identified by high performance liquid chromatography (HPLC) and HPLC-Q-TOF-MS/MS. Proliferation and migration experiments in vitro as well as tumor growth in vivo were performed to evaluate NPs on triple negative breast cancer (TNBC). ResultsThe present article quickly discovered the key active ingredients from SH herb pair70%SH-NPs, which were mainly composed of aurantiamide acetate (from HD), scutebarbatine A (from SB) and palmitin (from HD). 70%SH-NPs significantly inhibited proliferation and migration of breast cancer cells in vitro together with tumor growth in nude mice. ConclusionsThe results maybe explain the wisdom of SH herb pair compatibility, that is, the active ingredients of the two single drugs self-assemble to form NPs via supramolecular chemical bonds during the decoction process. In summary, the present article might inspire the studies of the relationship between active phytochemicals and NPs in herbs, and the corresponding pharmacological implications.

Co-immobilized Alcohol Dehydrogenase and Glucose Dehydrogenase with Resin Extraction for Continuous Production of Chiral Diaryl Alcohol.

Ni2+-functionalized porous ceramic/agarose composite beads (Ni-NTA Cerose) can be used as carrier materials to immobilize enzymes harboring a metal affinity tag. Here, a 6×His-tag fusion alcohol dehydrogenase Mu-S5 and glucose dehydrogenase from Bacillus megaterium (BmGDH) were co-immobilized on Ni-NTA Cerose to construct a packed bed reactor (PBR) for the continuous synthesis of the chiral intermediate (S)-(4-chlorophenyl)-(pyridin-2-yl) methanol ((S)-CPMA) NADPH recycling, and in situ product adsorption was achieved simultaneously by assembling a D101 macroporous resin column after the PBR. Using an optimum enzyme activity ratio of 2:1 (Mu-S5: BmGDH) and hydroxypropyl-β-cyclodextrin as co-solvent, a space-time yield of 1560 g/(L·d) could be achieved in the first three days at a flow rate of 5 mL/min and substrate concentration of 10 mM. With simplified selective adsorption and extraction procedures, (S)-CPMA was obtained in 84% isolated yield.

Preparation and characterization of a low-phenylalanine whey hydrolysate using two-step enzymatic hydrolysis and macroporous resin adsorption

A low-phenylalanine (Phe) diet is indispensable for phenylketonuria patients. The objectives of this study were to develop a two-step enzymatic method to effectively release Phe from whey protein, and then to prepare a low-Phe hydrolysate (LPH) by using adsorbent. Thermolysin (T) or Pancreatin (P) combined with Protease A2SD (A2) showed the degree of hydrolysis ranging from 42.62% to 45.43% and released more than 90% Phe from whey protein. The D101 macroporous resin column removed 98.38% Phe from TA2 hydrolysate (TA2H), compared to 64.50% of Phe removal by activated carbon. The protein content of LPH prepared by a magnified D101 resin column was 60.41 g/100 g hydrolysate and its Phe content was 1.38 ± 0.11 mg/g protein equivalent. After adsorption, the percentage of peptides below 600 Da significantly increased from 50.79% to 93.60%, while the intensity of bitterness and astringency measured by electronic-tongue also increased. However, aroma-active compounds such as Millard reaction and lipid oxidation products were reduced after absorption. The LPH prepared in our study could be a suitable protein substitute to make phenylketonuria food.

Optimization of extraction for Anemarrhena asphodeloides Bge. using silica gel-based vortex-homogenized matrix solid-phase dispersion and rapid identification of antioxidant substances.

A novel and simple method was established for the extraction and determination of seven compounds in Anemarrhena asphodeloides Bge. using silica gel-based vortex-homogenized matrix solid-phase dispersion and ultra-high performance liquid chromatography quadrupole-time of-flight mass spectrometer. The conditions for the extraction were optimized. Silica gel was used as the dispersant, 50% methanol-water was selected as an elution solvent and the grinding time was 3min. Compared with the traditional ultrasonic-assisted extraction, the developed method was rapid and efficient. In order to screen potential antioxidants, extract dealing with the optimized method was applied to a polyamide chromatography column and a D-101 macroporous resin column. Fr.2.2 showed the highest antioxidant activities with the most content of flavonoid. A total of 25 peaks were identified from the active fraction. A 2,2'-diphenyl-1-picrylhydrazyl ultra-high performance liquid chromatography coupled with mass spectrometry approach was adopted for the rapid and exact screening and identification of antioxidant compounds. It indicated that flavonoids exhibited potential antioxidant activities. The antioxidant activities of nine monomeric compounds in vivo were tested. Structure-activity relationships were discussed. Five flavonoids with the concentration of 500 µg/mL would reduce the oxidative stress of PC12 cells that were induced with 2,2'-azobis[2-methylpropionamidine] dihydrochloride.

Investigation of Chemical Composition, Antioxidant Activity, and the Effects of Alfalfa Flavonoids on Growth Performance.

The flavonoids were extracted from alfalfa using ethanol assisted with ultrasonic extraction and purified by D101 macroporous resin column chromatography. The chemical composition and content of ethanol elution fractions (EEFs) were assessed by ultrahigh-performance liquid chromatography and hybrid quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS) and aluminum nitrate-sodium nitrite-sodium hydroxide colorimetric method. The in vitro antioxidant activity of two EEFs was conducted by scavenging DPPH free radical, and the main antioxidants of 75% EEFs were screened using DPPH-UHPLC. Moreover, the in vivo antioxidant activity of 75% EEFs and the growth performance of broilers were studied. The results showed that the content of 30% and 75% EEFs was 26.20% and 62.57%. Fifteen compounds were identified from 75% EEFs, and five of them were reported in alfalfa for the first time. The scavenging activity of 75% and 30% EEFs (200 μg/mL) against DPPH was 95.51% and 78.85%. The peak area of 5,3′,4′-trihydroxyflavone and hyperoside was decreased by 82.69% and 76.04%, which exhibited strong scavenging capacities. The total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) level of three treated groups against the normal control group (NC) fed with basal diet significantly increased by 3.89-24.49%, 0.53-7.39%, and 0.79-11.79%, respectively. While the malondialdehyde (MDA) decreased by 0.47-18.27%. Compared with the NC, the feed to gain ratio (F : G) of three treated groups was lowered by 2.98-16.53% and survival rate of broilers significantly increased. Consequently, 75% EEFs extracted from alfalfa exhibited powerful antioxidant activities and might be a potential feed additive to poultry and livestock.

Screening potential α-glucosidase inhibitors from Anemarrhena asphodeloides using response surface methodology coupled with grey relational analysis

ABSTRACT Objective To screen potential α-glucosidase inhibitors from Anemarrhena asphodeloides. Methods Response surface methodology employing Box-Behnken design was used to optimize conditions for the extraction of α-glucosidase inhibitory active compounds from A. asphodeloides. The powders (20.0 g) of A. asphodeloides were extracted under the optimized conditions. The extract was applied to a D-101 macroporous resin column. It was eluted with ethanol and water to give six fractions. Compounds from the active fraction were identified by UPLC-Q-TOF-MS. The structure-activity relationship was discussed based on grey relational analysis. Results The optimum extraction conditions were as follows: ethanol concentration, 100%; extraction temperature, 51 °C; and solvent to solid ratio, 23 mL/g. It indicated that the active compounds were concentrated into 80% ethanol fraction. Twenty five steroid saponins from 80% ethanol fraction were identified by UPLC-Q-TOF-MS. Peaks 19 and 23 were tentatively identified as new structures. The predicted α-glucosidase inhibitory activities of the compounds were 7 > 2 > 1 > 22 > 23 > 3 > 9 > 21 > 24 > 4 > 13 > 8 > 14 > 16 > 17 > 25 > 6 > 19. Conclusion The fraction eluted by 80% ethanol showed the best inhibitory activity. After analyzing the data of UPLC-Q-TOF-MS, 25 steroid saponins were tentatively identified in this fraction.

Hypoglycemic activity and constituents analysis of blueberry (Vaccinium corymbosum) fruit extracts.

BackgroundTo investigate hypoglycemic activity and elucidate the active composition of the fruit blueberry (Vaccinium corymbosum).MethodsMethanol extracts of blueberry (MEB) were separated using a D101 macroporous resin column to yield quinic acid derivative (Fr.1)- and flavonoid (Fr.2)-rich fractions. The effects of the blueberry extracts on mRNA expression of GLUT-2 (glucose transporter type 2) and PPARγ (peroxisome proliferator-activated receptor-γ), as well as on the activities of PPRE (peroxisome proliferator response element) and NF-κB were analyzed in LO2 normal liver cells. Real-time PCR was used to detect the expression of GLUT-2, PPARγ, TNF-α, IL-1β, and IL-6 mRNA. The PPRE and NF-κB activities were detected by a luciferase reporter assay. Western blotting was used to detect the levels of PPARγ, GLUT-2, and p65. The active compositions were isolated using various chromatography columns, and were analyzed by NMR.ResultsmRNA and protein expression of GLUT-2 and PPARγ were significantly increased upon treatment with 400 μg/mL extracts of blueberry (P<0.05). The PPRE activity was also significantly increased in a dose-dependent manner upon administration of MEB (P<0.05). Furthermore, the NF-κB activity induced by lipopolysaccharides was inhibited by MEB (P<0.05). No fraction separated from MEB exhibited PPRE activation or NF-κB inhibition activity. Blueberry extract may execute its hypoglycemic activity by stimulating expression of GLUT-2 and PPARγ, and by inhibiting the inflammatory pathway. Together, quinic acid derivatives and flavonoids may result in a synergistic effect. Fourteen phenolic acids, including eight flavonoids, four quinic acid derivatives, and two other phenolic acids, were isolated and identified, and caffeoylquinic acid derivatives and quercetin glycosides were found to be the major constituents of blueberry.ConclusionBlueberry may have hypoglycemic activity that functions through synergistic effects with caffeoylquinic acid derivatives and quercetin glycosides.

CO2 supercritical fluid extraction and characterization of polysaccharide from bamboo (Phyllostachys heterocycla) leaves

CO2 supercritical fluid technique with an ethanol modifier was used to extract bamboo (Phyllostachys heterocycla) leave’s polysaccharide (BLPs) regarding various operating conditions. Pressure (30, 40 and 50 MPa), extraction temperature (40, 45 and 50 °C), extraction time (2, 3 and 4 h) and modifier dosage (20, 25 and 30 mL) were important operating factors affecting the yield and total antioxidant activity of each BLPs extraction. The optimal parameters for harvesting the BLPs were an extracting time of 2 h plus 40 MPa pressure with 30 mL of ethanol modifier at 50 °C. In this case, 2.47% BLPs (in wt) was extracted. The BLPs, after purified by D101 macroporous resin and DEAE-cellulose column, with the average molecular weight of 5.05 × 104 Da are composed of rhamnose, fucose, arabinose, xylose, mannose, glucose, and galactose in the molar ratio of 1.318:2.100:1.189:1.016:1:4.776:1.318, respectively. It showed the best antioxidant activity, and its total antioxidant activity of the BLPs extracted was equivalent to 20.70 mg ascorbic acid [(VcE)/g]. In a word, our technique should be potential for producing the best yield and activity of bamboo (Phyllostachys heterocycla) leave’s polysaccharide.

Extraction, purification and anti-radiation activity of persimmon tannin from Diospyros kaki L.f.

In this study, persimmon tannin was extracted from Diospyros kaki L.f. using ultrasound-assisted extraction and purified by D101 macroporous resin column chromatography and polysulfone ultrafiltration membrane. The tannin content of the final persimmon tannin extracts was attained to 39.56% calculated as catechin equivalents. Also, the radioprotective effects of persimmon tannin for HEK 293T cells proliferation and apoptosis after Gamma irradiation were investigated by CCK-8, Hoechst 33258 staining, flow cytometry assay and intracellular reactive oxygen species assay (ROS). Persimmon tannin was pre-incubated with HEK 293T cells for 12 h prior to Gamma irradiation. It was found that pretreatment with persimmon tannin increased cell viability and inhibited generation of Gamma-radiation induced ROS in HEK 293T cells exposed to 8 Gy Gamma-radiation. The percentage of apoptotic cells were only 6.7% when the radiation dose was 8 Gy and pretreated with 200 μg/ml of persimmon tannin. All these results indicated that persimmon tannin offered a potent radioprotective effect on cell vitality and cell apoptosis of Gamma-radiation exposure in HEK 293T cells. This study would serve as a pre-clinical evaluation of persimmon tannin for use in people with radiation protection.

A New Monoterpenoid Indole Alkaloid from Tabernaemontana corymbosa

Tabernaemontana corymbosa is used in folk medicine for the treatment of hypertension, sore throat, and abdominal pain by the Dais in Yunnan Province. The crude alkaloid extract of T. corymbosa were successively subjected to D-101 macroporous resin, MCI gel, and ODS column chromatography and purified via preparative HPLC to afford a new aspidospermar-type monoterpenoid indole alkaloid, named tabercarpamine K (1). The structure of 1 was elucidated using spectroscopic methods. Compared to the structure of vincamine, a hydroxyl group is substituted at C-19 and a proton, instead of a hydroxyl, is attached to C-16 in 1.

Elution-extrusion counter-current chromatography for the separation of two pairs of isomeric monoterpenes from Paeoniae Alba Radix.

In this work, a simple and efficient protocol for the rapid separation of two pairs of isomeric monoterpenes from Paeoniae Alba Radix was developed by combining macroporous resin and elution-extrusion counter-current chromatography. The crude extract was firstly subjected to a D101 macroporous resin column eluted with water and a series of different concentrations of ethanol. Then, effluents of 30 and 95% ethanol were collected as sample 1 and sample 2 for further counter-current chromatography purification. Finally, a pair of isomers, 96 mg of compound 1 and 48 mg of compound 2 with purities of 91.1 and 96.2%, respectively, was isolated from 200 mg of sample 1. The other pair of isomers, 14 mg of compound 3 and 8 mg of compound 4 with purities of 93.6 and 88.9%, respectively, was isolated from 48 mg of sample 2. Their purities were analyzed by high-performance liquid chromatography, and their chemical structures were identified by mass spectrometry and (1) H NMR spectroscopy. Compared to a normal counter-current chromatography separation, the separation time and solvent consumption of elution-extrusion counter-current chromatography were reduced while the resolutions were still good. The established protocol is promising for the separation of natural products with great disparity of content in herbal medicines.

Chemical Constituents of the Roots of Triosteum pinnatifidum

The genus Triosteum Linn., belonging to the family Caprifoliaceae, consists of about eight species mainly distributed in East Asia and North America [1]. In traditional Chinese medicine the roots of T. pinnatifidum Maxim., locally known as “Tian Wang Qi,” are used for the treatment of rheumatic lumbocrural pain, bruises, dyspepsia, and menoxenia [2]. Ten iridoids have been reported from the title plant, which were the characteristic constituents of the Triosteum genus [3]. In continuation of our studies on chemical constituents from the roots of T. kirilowii, 17 compounds were isolated and elucidated as loganic acid (1), epi-vogeloside (2), 7-O-butylsecologanic acid (3), foliasacioside B1 (4), 3,5-di-O-caffeoyl quinic acid (5), methyl 3,5-di-O-caffeoyl quinate (6), 4,5-di-O-caffeoyl quinic acid (7), methyl 4,5-di-O-caffeoyl quinate (8), 4,5-di-O-caffeoyl quinic acid butyl ester (9), chlorogenic acid butyl ester (10), caffeic acid (11), ethyl caffeate (12), ursolic acid (13), betulin (14), -sitosterol (15), daucosterol (16), and daucosterol-6 -O-palmitate (17). To the best of our knowledge, all these compounds were obtained from the genus Triosteum for the first time. General. 1H and 13C NMR spectra were obtained on Varian INOVA 500 MHz (Varian, USA) or Bruker Avance DRX 400 MHz spectrometers (Bruker, Switzerland). All solvents used were of analytical grade (Tianjin Jiangtian Chemical Technology Co. Ltd., China). Silica gel (100–200 and 200–300 mesh, Qingdao Haiyang Chemical Co. Ltd., China), Sephadex LH-20 gel (Amersham Pharmacia Biotech AB, Sweden), LiChroprep RP-18 (40–63 m, Merck, Germany), and resin D101 (Tianjin Haiguang Chemical Co. Ltd., China) were used for column chromatography. TLC was carried out using precoated plates with GF254 silica gel (Qingdao Haiyang Chemical Co. Ltd., China). Spots on TLC were visually observed under UV light and by spraying with 5% H2SO4 reagent followed by heating. Plant Material. The roots of T. pinnatifidum were collected from Mei County, Shaanxi Province of China in August 2006, and authenticated by Prof. Zhen-Hai Wu, Northwest A F University. A voucher specimen (S200608007) was deposited in the herbarium of School of Pharmaceutical Science and Technology, Tianjin University, China. Extraction and Isolation. The air-dried powdered roots (20 kg) were refluxed with 95% EtOH and then with 60% EtOH twice. The extract was suspended in water and successively partitioned with petroleum ether (60–90 C), CHCl3, EtOAc, and n-BuOH in turn. The petroleum ether extract (80 g) was subjected to silica gel column chromatography eluting with petroleum ether–EtOAc (98:2 7:3) in a gradient manner to afford compounds 14 (20 mg) and 15 (2 g). The EtOAc extract (260 g) was subjected to silica gel column chromatography eluting with CH2Cl2–MeOH (97:3 8:2) in a gradient manner, and 88 fractions were collected. Fractions 16–18 were chromatographed over a silica gel column repeatedly and recrystallized with Me2CO to afford compound 13 (60 mg). Fractions 19–29 were subjected to silica gel column chromatography and further purified by Sephadex LH-20 to yield compound 12 (50 mg). Fractions 30–36 were purified by silica gel column chromatography and recrystallized to give compound 17 (80 mg). Fractions 49–55 were recrystallized with Me2CO to obtain compound 16 (560 mg). The n-BuOH extract (1.4 kg) was chromatographed over a D101 macroporous resin column eluting with H2O and 30%, 50%, 70%, and 95% EtOH. The 30% EtOH eluate (400 g) was subjecred to silica gel column chromatography eluting with EtOAc–MeOH (95:5 6:4) in a gradient manner. Fractions 1–2 were subjected to silica gel column chromatography and

The effects of extract ofWujia-Shenghua capsule on proliferation of the MCF-7 cells

ObjectiveTo observe the effect of estrogenic-like effects ofWujia-Shenghua capsule and effective medication site.MethodsThrough D-101 macroporous resin column methods,Wujia-Shenghua capsule with 60% ethanol extraction was separated for water elution part, 20% ethanol elution part, 40% ethanol elution part, 60% ethanol elution part. Then each elution part was respectively mixed into concentration of 10mg/ml,1mg/ml, 0.1mg/ml, 0.01 mg/ml and acted on the MCF-7 cell to have, MTT test and rate of PR calculation.Results Compared with the blank control group（100%）, when the concentration was 0.1 mg/ml, water elution part, 20% ethanol elution part and 60% ethanol elution part（98.10%, 101.06%, 106.04%）had no effect on the proliferation of MCF-7 cells, there was not statistically significant（P〉0.05）, while the 40% ethanol elution part（108.22%）can promote the proliferation of MCF-7 cells,there was a statistically significant（P〈0.05）. When the concentration was 1 mg/ml, 20%, 40% and 60% ethanol elution part（111.72%, 122.48%, 115.35%）can distinctly promote the proliferation of MCF-7 cells, there was a significant difference between four groups（P〈0.01）.ConclusionThe 40% ethanol elution part ofWujia-shenghua capsule has the strongest estrogen activity on plant. Key words: Wujia-Shenghua capsule extract MCF-7 cell MTT test Plant estrogen-like effects Effective medication site

Chemical Constituents of the Roots of Trichosanthes kirilowii

The genus Trichosanthes Linn. (Cucurbitaceae) is composed of about 50 species distributed mainly in Southeast Asia. In traditional Chinese medicine the roots of Trichosanthes kirilowii Maxim., commonly known as “Tian Hua Fen,” are used for the treatment of cough, inflammation, polydipsia, and diabetes [1]. Most previous phytochemical studies of T. kirilowii have focused on the fruits and seeds, and triterpenoids, sterols, amino acid, lipids, and other compounds have been isolated from different parts of the plant [2–5]. In our research on chemical constituents of the roots of T. kirilowii, which has rarely been reported in recent years, 18 compounds were isolated and elucidated as 2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid (1), allantoin (2), nicotinic acid (3), xanthine (4), adenine (5), uracil (6), thymine (7), guanosine (8), uridine (9), phenylalanine (10), tyrosine (11), 1-C-(p-hydroxyphenyl)-glycerol (12), furoic acid (13), p-hydroxybenzoic acid (14), p-hydroxybenzaldehyde (15), succinic acid (16), -spinasterol-3-O-D-glucopyranoside-6 -O-palmitate (17), and bryonolic acid (18). Plant Material. The roots of Trichosanthes kirilowii were collected from Hebei Province of China in July 2010 and authenticated by Prof. Tian-Xiang Li, Tianjin University of Traditional Chinese Medicine. A voucher specimen was deposited in the herbarium of pharmacognosy, Tianjin University of Traditional Chinese Medicine. Extraction and Isolation. The air-dried roots (14 kg) were cut into small pieces and refluxed with 95% EtOH and then with 50% EtOH twice. The extract was suspended in water and successively partitioned with petroleum ether (60–90 C), CH2Cl2, EtOAc, and n-BuOH in turn. The petroleum ether extract (45 g) was subjected to silica gel column chromatography eluting with petroleum ether–EtOAc in a gradient manner, and further purified by silica gel and Sephadex LH-20 column chromatography repeatedly to afford compounds 17 (80 mg) and 18 (25 mg). The CH2Cl2 extract (30 g) was subjected to silica gel column chromatography eluting with gradient CH2Cl2–EtOAc to get compounds 12 (8 mg) and 15 (6 mg). The EtOAc extract (18 g) was repeatedly separated by silica gel, Sephadex LH-20, and preparative TLC to yield compounds 2 (85 mg), 3 (5 mg), 4 (5 mg), 6 (72 mg), 9 (3 mg), 13 (7 mg), 14 (15 mg), and 16 (100 mg). The n-BuOH extract (170 g) was chromatographed over a D101 macroporous resin column eluting with H2O and 30%, 50%, 70%, and 95% EtOH. The 30% eluate (37 g) was subjected to silica gel column chromatography and further purified on Sephadex LH-20 and ODS columns to give compounds 1 (20 mg), 5 (20 mg), 7 (3 mg), 8 (150 mg), 10 (80 mg), and 11 (8 mg). The structures of the compounds were identified by a combination of spectroscopic methods (MS, 1H NMR, and 13C NMR) and physicochemical properties. The spectroscopic data of all the compounds were in good agreement with the literature data. 1H and 13C NMR spectra were recorded on 400 MHz and 600 MHz Bruker AV III spectrometers (Bruker, Switzerland). ESI-MS data were obtained on a Waters Quattro Premier XE mass spectrometer (Waters, USA). Silica gel (100–200 and 200–300 mesh, Qingdao Haiyang Chemical Co. Ltd., China), Sephadex LH-20 gel (GE Healthcare Bio-Sciences AB, Sweden), ODS (YMC Co. Ltd, Japan), and D101 resin (Tianjin Haiguang Chemical Co. Ltd., China) were used for column chromatography. TLC was carried out using precoated plates with GF254 silica gel (Qingdao Haiyang Chemical Co. Ltd., China). Spots on TLC were visually observed under UV light and by spraying with 5% H2SO4 reagent followed by heating.

New lignans from the bioactive fraction of Sambucus williamsii Hance and proliferation activities on osteoblastic-like UMR106 cells

Four new lignans (1, 7–9), together with nine known ones, were isolated from the anti-osteoporosis fraction of the extract of Sambucus williamsii Hance which was eluted by 50% and 95% aqueous ethanol over D101 macroporous resin column. Their structures were elucidated by NMR spectroscopic analyses, and the absolute configurations of all compounds were determined by application of circular dichroism method. All the compounds were reported for the first time from the Sambucus genus and firstly studied for their proliferation effects on osteoblastic-like UMR 106 cell. The data showed that compounds 2–9 significantly promoted cell proliferation in some dose, especially compounds 2, 3, 4, 5, and 7 increased osteoblastic cell numbers by 31.3%, 28.3%, 25.6%, 25.1% and 26.0% at 10−10M, 10−10M, 10−7M, 10−10M and 10−10M, respectively, which suggested that lignans were the components accounting for the bone protective effects of SWH.

The effects of Zibu Piyin Recipe components on scopolamine-induced learning and memory impairment in the mouse

Ethnopharmacological relevanceThe Zibu Piyin Recipe (ZBPYR) is derived from Zicheng Decoction, a traditional Chinese medicine formula recorded in the book of Bujuji, written by Wu Cheng in the Qing dynasty and used for clinical treatment of amnesia. Our aim was to study the effects of Zibu Piyin Recipe (ZBPYR) fractions on scopolamine-induced learning and memory impairment in the mouse. Materials and methodsCrude extracts were prepared using various solvents, and individual fractions produced following D101 macroporous resin column chromatography. The passive avoidance task, step down test and Morris water maze test were then performed in mice for the evaluation of learning and memory alterations. The effective fractions were then analyzed using GC–MS and polysaccharide measurement methods, respectively. ResultsThe treatment group latency for the alcohol precipitation from water part (EP) and 95% ethanol part (95%E) following D101 macroporous resin column chromatography was significantly prolonged when compared to that of the scopolamine treated groups for both the passive avoidance task and step down test. In the Morris water maze tests, treatment with EP and 95%E resulted in a significantly shorter escape latency time (from the fourth day and the second day) and swimming distance (on the third day and from the third day) in scopolamine-induced mice. In the memory retention test, treatment with EP and 95%E dramatically shortened the latency to cross platform location and increased the numbers of platform location crosses in the scopolamine-induced mice. The polysaccharide content in EP was determined to be 69.79%. The 95%E was found to mainly contain asarone, α-cadinol, isocalamendiol, 2,4,7,14-tetramethyl-4-vinyl-tricyclo[5.4.3.0(1,8)]tetradecan-6-ol, 3-isopropyl-6,7-dimethyltricyclo[4.4.0.0(2,8)]decane-9,10-diol, 2-methyl-9-(prop-1- -en-3-ol-2-yl)-bicyclo[4.4.0]dec-2-ene-4-ol, diepicedrene-1-oxide, 7-methoxy-6-(3- -methyl-2-oxobutyl)-2H-1-benzopyran-2-one and diisooctyl phthalate when assessed using GC–MS analysis. ConclusionThese findings suggest that the polysaccharide and volatile oil present in ZBPYR exhibit ameliorating effects on scopolamine-induced memory dysfunction.