Preparative Isolation and Purification of Altertoxin I from an Alternaria sp. by HSCCC

Altertoxin I (ATX I) is one of the common mycotoxins produced by genus Alternaria which is a common food pathogen of fruits and grains. To prepare enough quantity of pure ATX I for further research of mutagenicity and toxicology tests, a novel method using preparative high-speed counter-current chromatography (HSCCC) was developed. The ethyl acetate crude extracts of the acetone washes obtained after fermentation of Alternaria sp. was separated using a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–water (2:5:5:6, v/v). Collected fractions were analyzed by LC and identified by EI–MS and NMR analysis. The technique can isolate mg levels of the target compound per run.     

Rapid screening,identification, and purification of neuraminidase inhibitors from Lithospermum erythrorhizon Sieb.et Zucc. by ultrafiltration with HPLC–ESI‐TOF‐MS combined with semipreparative HPLC

We put forward an efficient strategy based on bioassay guidance for the rapid screening, identification, and purification of the neuraminidase inhibitors from traditional Chinese medicines, and apply to the discovery of anti‐influenza components from Lithospermiun erythrorhizon Sieb.et Zucc. Ultrafiltration with high‐performance liquid chromatography and electrospray ionization time‐of‐flight mass spectrometry was employed for the rapid screening and preliminarily identification of anti‐influenza components from Zicao. Semipreparative high‐performance liquid chromatography was used for the rapid separation and purification of the target compounds. NMR spectroscopy, mass spectrometry, and UV spectroscopy were used for further structural identification, and the activity of the compounds was verified by in vitro assay. Five compounds were found to have neuraminidase inhibitory activity by this method. Subsequently, the five compounds were separated by semipreparative high‐performance liquid chromatography with the purity over 98% for all of them by high‐performance liquid chromatography test. Combined with the NMR spectroscopy, mass spectrometry, and UV spectroscopy data, they were identified as alkannin, acetylalkannin, isobutyrylalkannin, β,β‐dimethylacryloylalkannin and isovalerylalkannin. The in vitro assay showed that all five compounds had good neuraminidase inhibitory activities. These results suggested that the method is highly efficient, and it can provide platform and methodology supports for the rapid discovery of anti‐influenza active ingredients from complex Chinese herbal medicines.     

An efficient combination of supercritical fluid extraction and high-speed counter-current chromatography to extract and purify (E)- and (Z)-diastereomers of α-asarone and β-asarone from Acorus tatarinowii Schott

In this study, supercritical fluid extraction (SFE) was used to extract essential oil from Acorus tatarinowii Schott under the pressure of 25 MPa and temperature of 35°C. Two (E)- and (Z)-diastereomers of α-asarone and β-asarone were separated and purified by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:0.2:1:0.3, v/v). The separation yielded a total of 39 mg of α-asarone and 463 mg of β-asarone from 1.0 g of essential oil in one-step separation with the purity of 98.9 and 99.1%, respectively, as determined by HPLC. The chemical structures of these compounds were identified by EI-MS, (1)H-, and (13)C-NMR.     

Use of pH-Zone Refining Countercurrent Chromatography for Preparative Separation of Fangchinoline and Tetrandrine from Stephania tetrandra

pH-Zone-refining countercurrent chromatography with a multilayer coil planet centrifuge has been successfully used for separation of fangchinoline and tetrandrine from crude extracts of Stephania tetrandra. The two target compounds were completely resolved by use of the two-phase solvent system petroleum ether (60–90 °C)–ethyl acetate–methanol–water 5:5:1:9 (v/v), with 10 mm triethylamine in the organic stationary phase and 5 mm hydrochloric acid in the aqueous mobile phase. Separation of 3.5 g sample yielded 126 mg fangchinoline (LC purity >93%) and 249 mg tetrandrine (LC purity >95%). The structures of the compounds were confirmed by use of electrospray ionization mass spectrometry and ¹H NMR.     

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.     

大孔阳离子交换树脂对井冈霉素的吸附研究

比较了5种大孔树脂与常用的001×4凝胶树脂对井冈霉素的静态和动态吸附的性能实验,并选择了其中几种有代表性的树脂,研究其对井冈霉素的吸附特性及其机理.采用正交试验筛选出具有较大吸附量的大孔强酸性阳离子树脂HD-8,并对其工艺条件进行了优化,其较优吸附能力为15.34mg/ml树脂h;同时考察了HD-8树脂的解吸行为,发现用1mol/L的氨水:80%乙醇=1:1洗脱率为98.8%,洗脱效果明显.     

Ionic liquid‐based ultrasound‐assisted extraction of fangchinoline and tetrandrine from Stephaniae tetrandrae

An ionic liquid‐based ultrasound‐assisted extraction method has been developed for the effective extraction of fangchinoline and tetrandrine from Stephaniae tetrandrae. The effects of some ultrasound‐assisted extraction parameters including the concentration of [BMIM][BF₄], pH, ultrasonic power and time were investigated to optimize the ultrasound‐assisted extraction conditions. Compared to the regular ultrasound‐assisted extraction and traditional refluent extraction, the proposed [BMIM][BF₄]‐based ultrasound‐assisted extraction offered shorter extraction times (from 6 h to 40 min) and remarkable higher efficiencies (approximately 30% improved), which supported the suitability of the proposed approach. In addition, the proposed approach was confirmed by the good correlation coefficient (R²), recovery and reproducibility (RSD, n = 5), which were in the range of 0.9992–0.9995, 85.5–101.1%, and 1.87–4.33%, respectively.     

A Supramolecular-Based Dual-Wavelength Phototherapeutic Agent with Broad-Spectrum Antimicrobial Activity Against Drug-Resistant Bacteria

With the ever-increasing threat posed by the multi-drug resistance of bacteria, the development of non-antibiotic agents for the broad-spectrum eradication of clinically prevalent superbugs remains a global challenge. Here, we demonstrate the simple supramolecular self-assembly of structurally defined graphene nanoribbons (GNRs) with a cationic porphyrin (Pp4N) to afford unique one-dimensional wire-like GNR superstructures coated with Pp4N nanoparticles. This Pp4N/GNR nanocomposite displays excellent dual-modal properties with significant reactive-oxygen-species (ROS) production (in photodynamic therapy) and temperature elevation (in photothermal therapy) upon light irradiation at 660 and 808 nm, respectively. This combined approach proved synergistic, providing an impressive antimicrobial effect that led to the complete annihilation of a wide spectrum of Gram-positive, Gram-negative, and drug-resistant bacteria both in vitro and in vivo. The study also unveils the promise of GNRs as a new platform to develop dual-modal antimicrobial agents that are able to overcome antibiotic resistance.     

A Supramolecular‐Based Dual‐Wavelength Phototherapeutic Agent with Broad‐Spectrum Antimicrobial Activity Against Drug‐Resistant Bacteria

With the ever‐increasing threat posed by the multi‐drug resistance of bacteria, the development of non‐antibiotic agents for the broad‐spectrum eradication of clinically prevalent superbugs remains a global challenge. Here, we demonstrate the simple supramolecular self‐assembly of structurally defined graphene nanoribbons (GNRs) with a cationic porphyrin (Pp4N) to afford unique one‐dimensional wire‐like GNR superstructures coated with Pp4N nanoparticles. This Pp4N/GNR nanocomposite displays excellent dual‐modal properties with significant reactive‐oxygen‐species (ROS) production (in photodynamic therapy) and temperature elevation (in photothermal therapy) upon light irradiation at 660 and 808 nm, respectively. This combined approach proved synergistic, providing an impressive antimicrobial effect that led to the complete annihilation of a wide spectrum of Gram‐positive, Gram‐negative, and drug‐resistant bacteria both in vitro and in vivo. The study also unveils the promise of GNRs as a new platform to develop dual‐modal antimicrobial agents that are able to overcome antibiotic resistance.