Optimization of ultrasonic-assisted supramolecular solvent microextraction of coumarins from Cortex fraxini using response surface methodology combined with artificial neural network-genetic algorithm.

A simple, fast, and efficient ultrasonic-assisted supramolecular solvent microextraction combined with high performance liquid chromatography method was developed for the determination of coumarins in Cortex fraxini, including esculin, esculetin and fraxetin. In this study, a novel supramolecular solvent was prepared with 1-octanol, tetrahydrofuran and water for the first time, and its composition, viscosity, density, structure, and micromorphology were characterized. The prepared supramolecular solvent exhibited vesicular structures and had the characteristics of low viscosity. Through single-factor experiments, response surface methodology and artificial neural network-genetic algorithm, the optimal extraction conditions were obtained as follows: NaCl concentration of 1 mol mL-1, pH value of 10, solid-liquid ratio of 10:1, vortex time of 30 s, ultrasonic power of 100 W, ultrasonic temperature of 60 °C, ultrasonic time of 15 min, centrifugation speed of 5000 rpm, and centrifugation time of 1 min. The results demonstrated that the artificial neural network model exhibited maximum R-values of 0.98703, 0.97440, 0.99836, and 0.95447 for training, testing, validation, and all dataset, respectively. The minimum mean square errors were 0.75, 10.15, 1.99, and 2.63, respectively. This indicated that the predicted values were almost consistent with the actual values. Under the optimal conditions, the total extraction yields of target analytes reached 2.80 %. The calibration curves for each analyte exhibited excellent linearity within the linear range (r > 0.9993). The limits of detection and quantification ranged from 4.87 to 6.55 ng mL-1 and 16.24 to 21.84 ng mL-1, respectively. The recoveries ranged from 98.71 % to 111.01 % with relative standard deviations of less than 3.6 %. The present method had the advantages of short extraction time (15 min) and less solvent consumption (0.5 mL). The prepared supramolecular solvent was proved to have great potential in extracting coumarins from medicinal plants.

Polyphyllin II (PPII) Enhances the Sensitivity of Multidrug-resistant A549/DDP Cells to Cisplatin by Modulating Mitochondrial Energy Metabolism.

BACKGROUND/AIM: Cisplatin resistance often leads to treatment futility and elevated mortality rates in patients with lung cancer. One promising strategy to address this challenge involves the integration of traditional Chinese medicine (TCM) with chemotherapeutic drugs. Currently, the potential synergistic effect and underlying mechanism of polyphyllin II (PPII) and cisplatin combination in combating cisplatin (DDP) resistance in lung cancer remain unexplored. MATERIALS AND METHODS: In this study, we established a cisplatin resistance model using A549 cells and explored the underlying mechanisms of PPII in combination with cisplatin in A549/DDP resistant cells. Specifically, we assessed the impact of PPII combined with cisplatin on A549/DDP cell proliferation, viability, and the expression of apoptosis-related proteins. To gain deeper insights into the underlying mechanism, we examined the effects of PPII and cisplatin on mitochondrial function in A549/DDP cells. RESULTS: This combination induced cell cycle arrest at both the S phase and G2/M phase in A549/DDP cells, thereby promoting apoptosis. Western blotting confirmed that DDP acted synergistically with PPII to enhance the expression of apoptotic proteins, diminish the expression of anti-apoptotic proteins, and promote the expression of anti-proliferation proteins in the mitochondrial pathway of A549/DDP cells. CONCLUSION: The combination of PPII and cisplatin effectively modulated the mitochondrial function, thereby reversing drug resistance in A549/DDP cells. This innovative combination therapy shows significant promise as a novel strategy for overcoming cisplatin resistance in lung cancer.

Application of supramolecular solvent based on the surface-active ionic liquid in dispersive liquid-liquid microextraction of triazine herbicides in tea samples.

In this study, a novel supramolecular solvent based on surface-active ionic liquid was prepared and used as an extraction solvent for dispersive liquid-liquid microextraction of four triazine herbicides in tea samples. The formation mechanism, microstructure and physicochemical properties of supramolecular solvent were studied. Some parameters, including the molar ratio of surface-active ionic liquid to tetrahydrofuran, volume of supramolecular solvent, vortex time, pH of sample solution, type and amount of salt, were investigated and optimized. The good linearities (r > 0.9990) for the analytes were obtained. The limits of detection and quantification for triazine herbicides were in the range of 1.7-2.1 µg kg-1 and 5.6-7.1 µg kg-1, respectively. The spiked recoveries were 80.0-119.9 %. The supramolecular solvent prepared in this study has the advantages of simple preparation process, low viscosity and good dispersibility. It can be used for the extraction and enrichment of trace triazine herbicides in tea samples.

Analysis of the Primary and Post-Treatment Antibiotic Resistance of Helicobacter pylori in the Nanjing Area.

BACKGROUND: Resistance of Helicobacter pylori (H. pylori) to antibiotics is increasing worldwide. The study was aimed to understand the current situation of antibiotic resistance in Nanjing and to provide a reasonable basis for clinical selection of antibiotics to cure H. pylori. OBJECTIVE: To investigate the current status of H. pylori antibiotics resistance in the Nanjing area, and analyze the primary and post-treatment antibiotic resistance of H. pylori in this area. METHODS: During the period from July 2017 to December 2019, 1533 gastric mucosal specimens from patients with positive H. pylori confirmed by a breath test or rapid urease test were collected for isolation and identification of H. pylori. The agar dilution method was used for the antibiotic resistance test. RESULTS: The result showed that the resistance rates of H. pylori to amoxicillin, clarithromycin, levofloxacin, furazolidone, tetracycline and metronidazole were 2.74%, 47.03%, 33.59%, 0.91%, 0.52% and 80.76%, respectively in the period of July 2017 to December 2019. The resistance rates of H. pylori (primary vs. post-treatment) to amoxicillin, clarithromycin, levofloxacin, furazolidone, tetracycline and metronidazole were 1.83% vs. 6.08%, 38.62% vs. 77.81%, 27.41% vs. 56.23%, 0.58% vs. 2.13%, 0.33% vs. 1.22%, 78.57% vs. 88.75%, respectively. CONCLUSION: Antibiotic resistance of H. pylori remained a problem for the effective eradication of this pathogen and its associated diseases in the Nanjing area. For post-treatment eradication patients, clinicians should take into account regional antibiotic resistance rate, personal antibiotic exposure history, economic benefit ratio, adverse antibiotic reactions, antibiotic availability and other aspects.

Ionic liquid-based salt-induced liquid-liquid extraction of polyphenols and anthraquinones in Polygonum cuspidatum.

Ionic liquid-based salt-induced liquid-liquid extraction was developed for the first time and applied to the extraction of four active constituents, including polydatin, resveratrol, emodin, and physcion in Polygonum cuspidatum (P. cuspidatum). In this study, ionic liquid was used as extraction solvent. The dried P. cuspidatum samples purchased from the pharmacy were triturated and passed through a 120-mesh sieve. The obtained sample powders were dried to constant weight at 55 ℃, and then mixed with extraction solvent. The extraction was carried out with the aid of ultrasound. Three phases, including ionic liquid-rich, salt-rich and solid sample phases were formed in the presence of salt. The target analytes were enriched in ionic liquid phase and then determined by high performance liquid chromatography. The experimental parameters, such as type and volume of ionic liquid, type and amount of salt, pH value of extraction medium, ultrasound power, ultrasound time and centrifugal condition, were optimized. The calibration curves showed good linear relationship (r > 0.9994). The limits of detection and quantification were in the range of 2.8-29.5 and 9.4-98.3 ng mL-1, respectively. The spiked recoveries were between 92.16% and 105.41%. Compared with hot reflux extraction and ultrasound-assisted extraction, the proposed method requires less extraction solvent and time. The present method can be applied to the determination of polyphenols and anthraquinones in P. cuspidatum.

Matrix Solid-Phase Dispersion Coupled with High-Performance Liquid Chromatography Diode Array Detection for Simultaneous Determination of Four Lipophilic Constituents from Salvia miltiorrhiza Bunge.

A simple, rapid and efficient method based on matrix solid-phase dispersion coupled with high-performance liquid chromatography was developed for determination of lipophilic constituents, including dihydrotanshinone, tanshinone I, cryptotanshinone and tanshinone II A in Salvia miltiorrhiza Bunge Box-Behnken design was employed for optimization of the extraction conditions of matrix solid-phase dispersion, including mass ratio of dispersant to sample, volume of elution solvent, and amount of cleanup reagent. The optimal experimental results were obtained using 0.27 g of acid alumina as dispersant, 13 mL of acetonitrile as elution solvent and 0.36 g of acid alumina as cleanup reagent. The target analytes was determined by high-performance liquid chromatography. The recoveries of tanshinones obtained by analyzing the spiked samples were from 83.81% to 93.74% and relative standard deviations from 2.87% to 6.83%. Matrix solid-phase dispersion integrated the extraction and cleanup into a single step, which provides the advantages of being simple, fast and convenient. Compared with other conventional methods, the present method consumed less time and less organic solvent. The results demonstrate that this method has potential for the determination of active constituents and the quality control of traditional Chinese medicine.

Microwave-assisted ionic liquid homogeneous liquid-liquid microextraction coupled with high performance liquid chromatography for the determination of anthraquinones in Rheum palmatum L.

The microwave-assisted ionic liquid homogeneous liquid-liquid microextraction (MA-IL-HLLME) coupled with high performance liquid chromatography with diode array detection (HPLC-DAD) was developed for the determination of anthraquinones, including aloe-emodin, emodin, chrysophanol and physcion in root of Rheum palmatum L. Several experimental parameters influencing the extraction efficiency, including amount of sample, type and volume of ionic liquid, volume and pH value of extraction medium, microwave power and extraction time, concentration of NH4PF6 as well as centrifugal condition were optimized. When 140µL of ionic liquid ([C8MIM][BF4]) was used as an extraction solvent, target analytes can be extracted from sample matrix in one minute with the help of microwave irradiation. The MA-IL-HLLME is simple and quick. The calibration curves exhibited good linear relationship (r>0.9984). The limits of detection and quantification were in the range of 0.015-0.026 and 0.051-0.088µgmL(-1), respectively. The spiked recovery for each analyte was in the range of 81.13-93.07% with relative standard deviations lower than 6.89%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with ultrasonic and heat reflux extraction. The results indicated that the present method can be successfully applied to the determination of anthraquinones in medicinal plant.

Dispersive micro-solid-phase extraction of herbicides in vegetable oil with metal-organic framework MIL-101.

Dispersive microsolid-phase extraction based on metal-organic framework has been developed and applied to the extraction of triazine and phenylurea herbicides in vegetable oils in this work. The herbicides were directly extracted with MIL-101 from diluted vegetables oils without any further cleanup. The separation and determination of herbicides were carried out on high performance liquid chromatography. The effects of experimental parameters, including volume ratio of n-hexane to oil sample, mass of MIL-101, extraction time, centrifugation time, eluting solvent, and elution time were investigated. The Student's t test was applied to evaluate the selected experimental conditions. The limits of detection for the herbicides ranged from 0.585 to 1.04 µg/L. The recoveries of the herbicides ranged from 87.3 to 107%. Our results showed that the present method is rapid, simple, and effective for extracting herbicides in vegetable oils.

Ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction high-performance liquid chromatography for determination of tanshinones in Salvia miltiorrhiza Bge. root.

The ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction has been developed and applied to the extraction of four tanshinones, including dihydrotanshinone, tanshinone I, cryptotanshinone and tanshinone IIA in Salvia miltiorrhiza Bge. root. High performance liquid chromatography was applied to the separation and determination of the analytes. The ionic liquid was used as extraction solvent and target analytes were extracted with help of ultrasound. Then, ion-pairing agent was added into the sample solution, which resulted in the formation of water-insoluble ionic liquid in the solution. The phase separation was performed by centrifugation. The extraction, concentration and purification of target analytes were performed simultaneously. The experimental parameters, including type and volume of ionic liquid, sample amount, the size of sample particle, pH value of extraction medium, extraction temperature, extraction time, amount of ion-pairing agent and centrifuging time, were investigated and optimized. The calibration curves showed good linear relationship (r>0.9997). The limits of detection and quantification were in the range of 0.052-0.093 and 0.17-0.31 µg mL(-1), respectively. The recoveries were between 70.45% and 94.23% with relative standard deviations lower than 5.31%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with UAE and HRE. There was no obvious difference in the extraction yields of active constitutions obtained by the three extraction methods.

Simultaneous determination of nucleosides and their bases in Cordyceps sinensis and its substitutes by matrix solid-phase dispersion extraction and HPLC.

Nine nucleosides and nucleobases, including uracil, adenine, thymine, uridine, adenosine, thymidine, cytidine, guanosine, and cordycepin in natural Cordyceps sinensis, cultured Cordyceps mycelia, and Cordyceps fruiting bodies were extracted by matrix solid-phase dispersion (MSPD) and determined by HPLC. The experimental conditions for the MSPD extraction were optimized. Florisil was used as dispersant, petroleum ether as washing solvent, and methanol as elution solvent. The Florisil-to-sample ratio was selected to be 4:1 and no additional clean-up sorbent was needed. The calibration curves had good linear relationships (r > 0.9997). The LOD and LOQ were in the range of 12～79 and 41～265 ng/mL, respectively. The intra- and interday precision were lower than 8.3%. The recoveries were between 61.5 and 93.2%. The present method consumed less sample compared with ultrasonic extraction and heating reflux extraction (HRE). The extraction yields obtained by using the present method are much higher than those obtained by UE and comparable to those obtained by HRE.