Separation and purification of quinolyridine alkaloids from seeds of Thermopsis lanceolata R. Br. by conventional and pH-zone-refining counter-current chromatography.

In this work, the preparative separation of quinolyridine alkaloids from seeds of T. lanceolata by conventional and pH-zone-refining counter-current chromatography. Traditional counter-current chromatography separation was performed by a flow-rate changing strategy with a solvent system of ethyl acetate-n-butanol-water (1:9:10, v/v) and 200 mg sample loading. Meanwhile, the pH-zone-refining mode was adopted for separating 2.0 g crude alkaloid extracts with the chloroform-methanol-water (4:3:3, v/v) solvent system using the stationary and mobile phases of 40 mM hydrochloric acid and 10 mM triethylamine. Finally, six compounds, including N-formylcytisine (two conformers) (1), N-acetycytisine (two conformers) (2), (-)-cytisine (3), 13-ß-hydroxylthermopsine (4), N-methylcytisine (5), and thermopsine (6) were successfully obtained in the two counter-current chromatography modes with the purities over 96.5%. Moreover, we adopted nuclear magnetic resonance and mass spectrometry for structural characterization. Based on the obtained findings, the pH-zone-refining mode was the efficient method to separate quinolyridine alkaloids relative to the traditional mode.

Extraction and purification of cis/trans asarone from Acorus tatarinowii Schott: Accelerated solvent extraction and silver ion coordination high-speed counter-current chromatography.

Acorus tatarinowii Schott is a traditional Chinese medicine used to treat memory and cognitive dysfunction. Because of their efficacy and lower toxic effects, research on α- and ß-asarone, the phytoconstituents, has attracted attention owing to their remarkable pharmacological activities. Silver ion coordination complexation high-speed counter-current chromatography was used to separate these isomers from A. tatarinowii extract, coupled with accelerated solvent extraction. Accelerated solvent extraction parameters were investigated with single-factor and orthogonal testing. A two-phase solvent system composed of n-hexane-ethyl acetate-ethanol-water (2:1:2:1, v/v) with 0.50 mol/L silver ions was selected for separation. From 2.0 g crude extract, 1.4 g of ß-asarone and 0.09 g of α-asarone were obtained with purities over 98% by sequential sample loading in 20 h. The isolated compounds were identified by electrospray ionization mass spectrometry, 1H and 13C NMR. Silver ions significantly increased the separation factor and retention of the stationary phase. The chromatographic behavior indicated that cis-configuration was more strongly complexed with the silver ion. This was further demonstrated with the help of computational analysis. In conclusion, the established method could be employed to separate other cis-trans or E/Z isomers that form coordination complexes.

Stable isotopes reveal the formation diversity of humic substances derived from different cotton straw-based materials.

Humic substances (HS) are essential in environment processes and carbon (C) sequestration in soils. In this study, organic materials such as cotton straw and its derived compost and biochar were added to the soil on a C-equivalent basis and incubated for 30 and 180 days in order to investigate the different forms of plant biomass derived C sequestration in HS. The C distribution in humic acid (HA), fulvic acid (FA), and humin (Hu) derived from organic materials was investigated using the 13C isotope method, while the catalase, sucrose, and ß-glucosidase activities were also determined. The results showed that C3 distribution of Hu derived from straw, compost and biochar increased from 40.94% to 67.12%, 74.47% and 80.75%, respectively. In addition, the increase of C3 distribution of HA or FA derived from straw, compost and biochar were 4.69%, 10.09% and 1.49%, respectively. There were significantly positive correlations between catalase, sucrase and ß-glucosidase activities and C3 derived HA and FA. The principal component analysis showed that catalase, sucrase and ß-glucosidase were explained mainly by the first principal component indicating a significant correlation. These findings suggest that straw, compost and biochar are mainly sequestrated in Hu. Comparatively, the straw and compost are more likely to contribute to the formation of HA and FA in soil, but biochar favors the Hu, which helps in soil C sequestration. The formation of HA and FA derived from organic materials was supported by catalase, sucrase and ß-glucosidase activities.

Flavonoid epimers from custard apple leaves, a rapid screening and separation by HSCCC and their antioxidant and hypoglycaemic activities evaluation.

Leaves of custard apple are widely used in many places as a popular dietary supplement for the treatment of diabetes. Flavonoids are known to have anti-diabetic activity. In this study, the main flavonoid epimers were separated. The crude extract was first screened by HPLC-DAD before and after incubation with DPPH method to evaluate the antioxidants. An efficient extraction method was employed to remove non-flavonoid components. Subsequently, five main flavonoids with two pairs of epimers including quercetin-3-O-robinobioside, rutin, quercetin-3-O-ß-D-glucoside, kaempferol-3-O-robinobioside, and kaempferol-3-O-rutinoside were successfully separated by high-speed counter-current chromatography with ethyl acetate/n-butanol/water (4:1:5, v/v) coupled with online-storage inner-recycling mode. The structures of the separated compounds were identified by spectral techniques. The purity of the separated flavonoid glycosides was over 98%, as determined by HPLC. The separated pure constituents were found to possess the antioxidant capacities following DPPH radical scavenging protocol. The compounds (1-3) exhibited better antioxidant activity. Furthermore, the glucose uptake of crude flavonoid extract had better results than the crude ethanol extract. The present study demonstrates that the efficacy of custard apple leaves in lowering glucose level, and antioxidant capacities of separated pure compounds probably appear to be predominantly responsible for hypoglycaemic properties on HepG2 cells.

A simple and efficient linear gradient coupled with inner-recycling high-speed counter-current chromatography mode for the preparative separation of flavonoid glycosides from leaves of custard apple.

In general, the simultaneous separation and isolation of compounds with a broad polarity range from natural products is a challenge by ordinary high-speed counter-current chromatography (HSCCC). Indeed, the complex solvent system screening methods limit the broader application of HSCCC. We describe herein a rapid and efficient linear gradient CCC (LGCCC) method that enables the separation of flavonoid glycosides with a broad range of KD values from custard apple leaves. Inner-recycling CCC (IRCCC) mode has been further applied for the separation of compounds with similar KD values. Similarly to binary gradient HPLC, the LGCCC mode is achieved by adjustment of the proportion between ethyl acetate (pump A) and n-butanol (pump B) in an ethyl acetate/n-butanol/water solvent system. Various separation factors have been investigated, including separation mode, rotation speed, flow rate, and sample loading. The IRCCC mode has been used for the secondary separation of two epimers with a simple ethyl acetate/water (1:1, v/v) solvent system. Finally, five main flavonoid glycosides have been successfully separated, namely quercetin-3-O-robinobioside (1, 4.8 mg) and rutin (2, 12.1 mg), quercetin-3-O-ß-d-glucoside (3, 4.2 mg), kaempferol-3-O-robinobioside (4, 9.6 mg), and kaempferol-3-O-rutinoside (5, 24.6 mg). The purities of the separated flavonoid glycosides were over 98%, as determined by HPLC. Our study indicates that a suitable combination of LGCCC and IRCCC modes is an effective strategy for separating flavonoid glycosides from custard apple leaves. The mathematical expression of the LGCCC was deduced to illuminate the separation mechanism. It may also be applied to obtain component fractions for the further screening of active compounds from complex natural products.

Novel ultrasonic-assisted vacuum drying technique for dehydrating garlic slices and predicting the quality properties by low field nuclear magnetic resonance.

A novel ultrasonic-assisted vacuum drying technique for dehydrating garlic slices to give high quality products was developed. Garlic slices were dried at 60 °C using four methods: ultrasonic-assisted vacuum drying (USVD), vacuum drying (VD), ultrasonic-assisted drying (USD), and convective drying (CD, the control with no vacuum or ultrasonic applied). Drying kinetics, water-content changes, and properties of the garlic slices were assessed. Univariate linear and partial-least-squares regression models were used to predict the properties from low-field nuclear magnetic resonance parameters. USVD gave the shortest drying time (180 min less than CD) and provided a better garlic color and texture, and allicin retention rate than the other methods. Higher correlations between low-field nuclear magnetic resonance parameters and quality properties were found by partial-least-squares regression (PLSR) than by univariate analysis, with the analysis results being credible. Overall, ultrasonic-assisted vacuum drying produced high-quality products with its properties predicted well by low-field nuclear magnetic resonance.

Rapid screening and purification of potential alkaloid neuraminidase inhibitors from Toddalia asiatica (Linn.) Lam. roots via ultrafiltration liquid chromatography combined with stepwise flow rate counter-current chromatography.

Toddalia asiatica (Linn.) Lam. is a medical plant traditionally used to treat coughs, fevers, and various diseases. Alkaloids are the main active ingredients in Toddalia asiatica (Linn.) Lam., but traditional methods for screening and separation are complex and labor-intensive. In this work, an efficient strategy was developed to rapidly screen, identify, and separate neuraminidase inhibitors from Toddalia asiatica (Linn.) Lam. Ultrafiltration, high performance liquid chromatography, and time-of-flight mass spectrometry were employed for rapid screening and identification of neuraminidase inhibitors. A two-phase solvent system comprising n-hexane/ethyl acetate/methanol/water (5:5:3:7, v/v) was then selected for separation by high-speed counter-current chromatography. A sample loading of 200 mg and a stepwise flow rate were achieved by increasing the flow rate from 2 to 4 mL/min after 4 h. Three main fluoroquinoline alkaloids (haplopine, skimmianine, and 5-methoxydictamnine) along with two coumarins were obtained via one-step separation and their structures were determined by mass spectrometry and nuclear magnetic resonance. In vitro assays revealed skimmianine with half-maximal inhibitory concentration of 16.2 ± 0.7 µmol/L was selected as the potential highest neuraminidase inhibitor. The results suggest that ultrafiltration high performance liquid chromatography-mass spectrometry combined with high-speed counter-current chromatography is efficient for the screening and isolation of neuraminidase inhibitors from complex natural products.

Effect of cotton straw-derived materials on native soil organic carbon.

Different types of crop straw and their derived biochars and compost treatments have huge potential for carbon sequestration to sustain crop productivity. In this study, cotton straw (straw), cotton straw-derived compost (compost) and cotton straw-derived biochar (biochar) with equivalent carbon (C) content were added to soil and incubated for 30 and 180 days. The C sequestration potential of these organic materials was determined by 13C isotope trace method. The structural characteristic of soil organic carbon (SOC) was analyzed by solid-state 13C NMR spectroscopy. The SOC concentration was measured by wet oxidation and dry combustion methods. The results showed that 50.84%, 41.03% and 38.55% of native SOC were replaced by biochar, compost, and straw, respectively. The carbohydrate C and methoxyl C contents were significantly higher in straw and biochar amendments respectively, while phenolic C and alkyl C were high in compost amendment and a higher proportion of aryl C occurred in biochar treatment. These findings revealed that straw material was easier to be decomposed, but compost and biochar showing better stability.

A novel method for the highly efficient biotransformation of genistein from genistin using a high-speed counter-current chromatography bioreactor.

Genistein, an important soybean isoflavone compound, has gained attention for its significant properties. Compared with the glycone form of genistin, low content of genistein limits the use in food and pharmaceutical fields. In this study, a novel bioreactor with high-speed counter-current chromatography (HSCCC) was built for the highly efficient biotransformation of genistein from genistin. The solvent system for the bioreactor was selected according to the K D values. The selected solvent system was evaluated by the enzyme activity of ß-glucosidase. An ethyl acetate/buffer solution was selected as the preferred solvent system for the HSCCC bioreactor. Optimum reactor parameters were selected according to the retention of the stationary phase. The HSCCC bioreactor was operated using different flow rates, and 2.0 mL min-1 was chosen as the optimal flow rate with a conversion rate of over 90% within 24 h; the novel bioreactor easily immobilized and recycled the enzyme and could be applied in the preparation of genistein.

Correction: A novel method for the highly efficient biotransformation of genistein from genistin using a high-speed counter-current chromatography bioreactor.

[This corrects the article DOI: 10.1039/C8RA10629K.].

Application of coordination agent in high-speed counter-current chromatography for the preparative separation and isolation ginkgolic acids from the sarcotesta of Ginkgo biloba L.

An efficient coordination high-speed counter-current chromatography method for the preparative separation of ginkgolic acids from the sarcotesta of Ginkgo biloba L was developed. The type, concentration, and mechanism of the coordination agent were investigated. Following the use of four types of metal salts including silver nitrate, copper chloride, ferric chloride, and aluminium nitrate, n-heptane/ethyl acetate/methanol/acetic acid 5:4:1:1, v/v with 0.20 mol/L silver nitrate as the coordination agent was chosen as the optimum two-phase solvent system. Five main ginkgolic acids including C13:0, C15:0, C15:1, C17:1, and C17:2 were successfully separated with purities greater than 98%. The sample loading was 500 mg, the flow-rate was 2.0 mL/min, rotation speed was 800 rpm and temperature was 20°C. The structures of the separated ginkgolic acids were identified by comparison with standard samples and electrospray ionization mass spectrometry. The introduction of coordination chemistry in high-speed counter-current chromatography is novel and effective for the preparative separation and isolation of ginkgolic acids from the sarcotesta of Ginkgo biloba L and could also be applied to separate compounds which form coordination bonds in other complex natural products.

Chemical Constituents from Scindapsus officinalis (Roxb.) Schott. and Their Anti⁻Inflammatory Activities.

One new monoterpene glycoside (1), one new phenyl glycoside (2), one new caffeoyl derivative (3), were isolated from Scindapsus officinalis (Roxb.) Schott., along with four known compounds (4⁻7). Structures of the isolated compounds were elucidated by extensive analysis of spectroscopic data, especially 2D NMR data and comparison with literatures. All isolates were evaluated for anti-inflammatory activity against nitric oxide (NO) production in vitro. Compounds 3 and 7 exhibited moderate inhibitory effects on NO production with IC50 values of 12.2 ± 0.8 and 18.9 ± 0.3 µM, respectively.

Alkaloids from Scindapsus officinalis (Roxb.) Schott. and their biological activities.

Two new alkaloid glycosides (1-2), two new amide alkaloids (3 and 6), and two new amine alkaloids (4 and 5), were isolated from Scindapsus officinalis (Roxb.) Schott., along with four known alkaloids (7-10). Structural elucidation of the isolated compounds was established by spectroscopic analysis, especially 2D NMR techniques and comparison with literatures. The absolute configurations of compounds 1 and 2 were determined by the electronic circular dichroism calculations. Compound 1 represent the first example of alkaloid glycoside found to possess an unprecedented 2,5-methanocyclopenta[b]azepine core. All isolates were evaluated for cytotoxic activity against MCF-7 human breast cancer cell and anti-inflammatory activity against nitric oxide (NO) production in vitro. Compounds 4, 6 and 8 demonstrated moderate cytotoxic effects against MCF-7 cell (IC50 of 11.6, 25.3, and 20.5 µM, respectively), and 1, 7 and 9 exhibited moderate inhibitory effects on NO production (IC50 of 19.8, 30.3, and 11.8 µM, respectively).

Highly Graphitized Carbon Coating on SiO with a π⁻π Stacking Precursor Polymer for High Performance Lithium-Ion Batteries.

A highly graphitized carbon on a silicon monoxide (SiO) surface coating at low temperature, based on polymer precursor π⁻π stacking, was developed. A novel conductive and electrochemically stable carbon coating was rationally designed to modify the SiO anode materials by controlling the sintering of a conductive polymer, a pyrene-based homopolymer poly (1-pyrenemethyl methacrylate; PPy), which achieved high graphitization of the carbon layers at a low temperature and avoided silicon carbide formation and possible SiO material transformation. When evaluated as the anode of a lithium-ion battery (LIB), the carbon-coated SiO composite delivered a high discharge capacity of 2058.6 mAh/g at 0.05 C of the first formation cycle with an initial Coulombic efficiency (ICE) of 62.2%. After 50 cycles at 0.1 C, this electrode capacity was 1090.2 mAh/g (~82% capacity retention, relative to the capacity of the second cycle at 0.1 °C rate), and a specific capacity of 514.7 mAh/g was attained at 0.3 C after 500 cycles. Furthermore, the coin-type full cell composed of the carbon coated SiO composite anode and the Li[Ni0.5Co0.2Mn0.3O2] cathode attained excellent cycling performance. The results show the potential applications for using a π⁻π stacking polymer precursor to generate a highly graphitize coating for next-generation high-energy-density LIBs.

Development of online-storage inner-recycling counter-current chromatography for the preparative separation of complex components of alkylphenols from sarcotesta of Ginkgo biloba L.

High-speed counter-current chromatography (HSCCC) is becoming an effective and non-absorptive separation method from natural products. Due to the insufficient separation efficiency, it is challenging to separate complex components, especially for compounds with similar K D values. In this study, a novel and effective online-storage inner-recycling CCC method was used to separate alkylphenols from the sarcotesta of Ginkgo biloba L. A two-phase solvent system of n-heptane/ethyl acetate/methanol/acetic acid (5 : 4 : 1 : 1, v/v) was used for HSCCC separation of 500 mg crude extracts. After the inner-recycling of two fractions coupled with pre-HPLC, five main ginkgolic acids (C13:0, C15:1, C17:2, C15:1, C17:1) coupled with bilobol (C15:1) and a mixture were obtained from a non-stop separation using a storage loop and two six-way valves. This novel method was also evaluated and predicted by formula derivation. This method could be an effective, rapid, and simple approach to separate alkylphenols from the sarcotesta of G. biloba.

Five new chromone glycosides from Scindapsus officinalis (Roxb.) Schott.

Five new chromone glycosides, officinalisides A (1), B (2), C (3), D (4) and E (5) were isolated from Scindapsus officinalis (Roxb.) Schott., along with six known chromone derivatives, 7-O-α-l-rhamnosyl-nereugenin (6), undulatoside A (7), drynachromoside A (8), drynachromoside B (9), 5,7-dihydroxy-2-methyl chromone(10), 5,7-dihydroxy-2-hydroxymethyl chromone (11). Structural elucidation of the isolated compounds was established by spectroscopic analysis, especially 2D NMR techniques and comparison with literatures. The isolates were evaluated for anti-inflammatory activities in a LPS-stimulated RAW 264.7 model using inhibition of nitric oxide (NO) production as an indicator. Compounds 2, 4 and 10 demonstrated potential anti-inflammatory activity with IC50 values of 16.1, 19.1, and 13.4µM, respectively, compared to the positive control dexamethasone.

Extraction and purification of five terpenoids from olibanum by ultrahigh pressure technique and high-speed countercurrent chromatography.

Five terpenoids, including two new ones, 3,7-dioxo-tirucalla-8,24-dien-21-oic acid (2) and 3α-acetoxyl-7-oxo-tirucalla-8,24-dien-21-oic acid (3), and three known ones, boscartol A (1), 11-keto-ß-boswellic acid (4), and acetyl-11-keto-boswellic acid (5), have been extracted by the ultrapressure extraction and purified by pH-zone-refining countercurrent chromatography and high-speed countercurrent chromatography from olibanum. For ultrapressure extraction, the optimal condition including 200 MPa of extraction pressure, ethyl acetate of extraction solvent, 1:20 (g/mL) of solid/liquid ratio, and 2 min of extraction time were obtained. For the separation, from 1.5 g of the terpenoid extract, 220.1 mg of 4, 255.5 mg of 5, and 212.3 mg of the mixture of 1, 2, and 3 were obtained by pH-zone-refining countercurrent chromatography under the solvent system of chloroform/ethyl acetate/methanol/water (3:1:3:2, v/v/v/v) with aqueous ammonia and trifluoroacetic acid as retention and eluter agents. The enriched mixture (210 mg) was further separated by conventional high-speed countercurrent chromatography with petroleum ether/ethyl acetate/methanol/water (1:0.8:1.1:0.6, v/v/v/v), yielding 30.1 mg of 1, 35.5 mg of 2, 12.3 mg of 3. The structures of these five terpenoids were elucidated by extensive spectroscopic methods.

A novel pyrazole-containing indolizine derivative suppresses NF-κB activation and protects against TNBS-induced colitis via a PPAR-γ-dependent pathway.

The nuclear factor-κB (NF-κB)-mediated activation of macrophages plays a key role in mucosal immune responses in Crohn's disease (CD). Moreover, increasing evidence shows that the activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) exerts satisfactory anti-inflammatory effects in experimental colitis models, mostly by suppressing NF-κB-mediated macrophage activation. Therefore, exploring therapeutic strategies to activate PPAR-γ and inhibit the NF-κB pathway in colonic macrophages holds great promise for the treatment of CD. In this study, five novel pyrazole-containing indolizine derivatives (B1, B2, B3, B4 and B5) were successfully synthesized and characterized, and their anti-inflammatory activities for CD treatment were also investigated. Among the five compounds, compound B4 effectively decreased the NF-κB-mediated production of the pro-inflammatory cytokine TNF-α in LPS-stimulated peritoneal macrophages. Moreover, compound B4 significantly ameliorated 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced mouse colitis symptoms, including body weight loss, colonic pathological damage and inflammatory cell infiltration. The results of western blotting and luciferase reporter assays indicated that compound B4 activated PPAR-γ and subsequently suppressed NF-κB activation. Conversely, the addition of the PPAR-γ antagonist GW9662 abrogated the anti-inflammatory effects of compound B4 both in vitro and in vivo. In summary, compound B4 activated the PPAR-γ pathway to inhibit downstream NF-κB signaling, which alleviated experimental colitis. Thus, this compound may serve as a potential therapeutic agent for patients with CD.

Preparation and Capacity-Fading Investigation of Polymer-Derived Silicon Carbonitride Anode for Lithium-Ion Battery.

Polymer-derived silicon carbonitride (SiCN) materials have been synthesized via pyrolyzing from five poly(silylcarbondiimide)s with different contents of carbon (labeled as 1-5#). The morphological and structural measurements show that the SiCN materials are mixtures of nanocrystals of SiC, Si3N4, and graphite. The SiCN materials have been used as anodes for lithium-ion batteries. Among the five polymer-derived SiCN materials, 5#SiCN, derived from dichloromethylvinylsilane and di-n-octyldichlorosilane, has the best cycle stability and a high-rate performance at the low cutoff voltage of 0.01-1.0 V. In lithium-ion half-cells, the specific delithiation capacity of 5#SiCN anode still remains at 826.7 mA h g-1 after 100 charge/discharge cycles; it can even deliver the capacity above 550 mA h g-1 at high current densities of 1.6 and 2 A g-1. In lithium-ion full cells, 5#SiCN anode works well with LiNi0.6Co0.2Mn0.2O2 commercial cathode. The outstanding electrochemical performance of 5#SiCN anode is attributed to two factors: (1) the formation of a stable and compact solid electrolyte interface layer on the anode surface anode, which protects the electrode from cracking during the charge/discharge cycle; and (2) a large amount of carbon component and the less Si3N4 phase in the 5#SiCN structure, which provides an electrochemical reactive and conductive environment in the SiCN structure, benefit the lithiation/delithiation process. In addition, we explore the reason for the capacity fading of these SiCN anodes.

A ZnS nanocrystal/reduced graphene oxide composite anode with enhanced electrochemical performances for lithium-ion batteries.

A simple route for the preparation of ZnS nanocrystal/reduced graphene oxide (ZnS/RGO) by a hydrothermal synthesis process was achieved. The chemical composition, morphology, and structural characterization reveal that the ZnS/RGO composite is composed of sphalerite-phased ZnS nanocrystals uniformly dispersed on functional RGO sheets with a high specific surface area. The ZnS/RGO composite was utilized as an anode in the construction of a high-performance lithium-ion battery. The ZnS/RGO composite with appropriate RGO content exhibits a high reversible specific capacity (780 mA h g-1), excellent cycle stability over 100 cycles (71.3% retention), and good rate performance at 2C (51.2% of its capacity when measured at a 0.1C rate). To further investigate this ZnS/RGO anode for practical use in full Li-ion cells, we tested the electrochemical performance of the ZnS/RGO anode at different cut-off voltages for the first time. The presence of RGO plays an important role in providing high conductivity as well as a substrate with a high surface area. This helps alleviate the typically problems associated with volume expansion and shrinkage during prolonged cycling. Additionally, the RGO provides multiple nucleation points that result in a uniformly dispersed film of nanosized ZnS that covers its surface. Thus, the high surface area RGO enables high electronic conductivity and fast charge transfer kinetics for ZnS lithiation/delithiation.