Exploration of chemical components and metabolite synthesis pathways in eight Ephedra species based on HS-GC-MS and UPLC-Q-TOF-MS.

Objective: Ephedra, widely used in clinical practice as a medicinal herb, belongs to the genus Ephedra in the family Ephedraceae. However, the presence of numerous Ephedra varieties and variants requires differentiation for accurate identification. Methods: In this study, we employed headspace gas chromatography mass spectrometry (HS-GC-MS), ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and global natural products social molecular networking (GNPS) for chemical component identification. Chemometric analysis was used to analyze the differential components. Metabolic analysis and Kyoto encyclopedia of genes and genomes (KEGG) enrichment were utilized to explore the synthesis pathways of different components. Result: A total of 83 volatile and 79 non-volatile components were identified in Ephedra species. Differential analysis revealed that among the eight Ephedra stems, 18 volatile and 19 non-volatile differential compounds were discovered, whereas Ephedra roots exhibited 21 volatile and 17 non-volatile markers. Volatile compounds were enriched in four synthetic pathways, while non-volatile components were enriched in five pathways among the differentiated components. Conclusion: This study is the first to conduct a comparative analysis of chemical components in different Ephedra species and parts. It provides a foundational reference for authenticating Ephedra herbs, evaluating medicinal resources, and comparing quality in future studies.

Chemical Characteristics and Comparison of Schizonepetae Herba and Schizonepetae Herba Carbonisata by Combination of GC-MS and UHPLC-MS Strategies.

BACKGROUND: Schizonepetae Herba (SH, Jingjie) and Schizonepetae Herba Carbonisata (SHC, Jingjie Tan) are two different forms of the same herbal material, with SHC being the processed product of SH. The different clinical efficacies of SH and SHC may be caused by changes in their chemical compositions. Despite this, there have been few studies that have reported on the comparative identification of SH and SHC. Therefore, the aims of this experiment are to investigate the differential changes of non-volatile and volatile components before and after SH processing. OBJECTIVES: To establish combination strategies for identifying the chemical markers in SH and SHC using ultra-HPLC-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) and headspace gas chromatography-mass spectrometry (HS-GC-MS). METHODS: UHPLC-Q-TOF-MS and HS-GC-MS methods was utilized to comprehensively discriminate between SH and SHC. To identify chemical markers, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were performed on 14 batches of SH and SHC. RESULTS: A total of 71 non-volatile compounds and 81 volatile compounds were tentatively identified in SH and SHC. Among these, 14 non-volatile compounds and 18 volatile oils were found to be potential characteristic markers that can differentiate between SH and SHC. CONCLUSION: The present work provides valuable information for understanding the chemical differences between SH and SHC. The results obtained from this research may serve as a scientific foundation for comprehensively revealing the mechanisms involved in the carbonizing processing method of stir-frying SH. HIGHLIGHTS: The chemical changes that occur before and after carbonizing SH were investigated using integrated methods based on LC-MS and GC-MS, and chemical markers in SH and SHC were identified.

Modulation of high-quality internal multifoci based on modified three-dimensional Fourier transform.

We report an efficient method to generate arbitrary three-dimensional (3D) parallel multifoci inside a material. Taking into account the numerical aperture of the objective lens and the refractive index of the material, the Ewald cap was modified with a longer radius, then the whole 3D intensity distribution inside the material could be calculated using only a single Fourier transform (FT). By introducing the adaptive weight coefficient, the uniformity of the 3D multifoci improves from 81.3% to 98.9%. By adjusting the axial resolution of the Ewald cap, the uniformity of the axial multifoci improves from 85.9% to 99.7%. In the experiment, we have realized one-dimensional (1D), 2D, and 3D structures inside the fused silica, which are in excellent agreement with the simulation results. The experimental results of the "H-U-S-T" structure demonstrate that customized arbitrary intensity distribution inside the material can be realized.

Optimization of the processing technology of schizonepetae herba carbonisata using response surface methodology and artificial neural network and comparing the chemical profiles between raw and charred schizonepetae herba by UPLC-Q-TOF-MS.

In this study, response surface methodology (RSM) and artificial neural network (ANN) were used to predict and validate the optimal processing method of Schizonepetae Herba Carbonisata (SHC). The highest overall desirability (OD) value of the total flavonoids content (TFC), total tannin content (TTC), and adsorption capacity (AC) were used as response values. The optimal processing technology processing time lasted 10 min at a processing temperature of 178 °C and the herbs/machine had a volume of 77 g/5 L. The Ultra Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry (UPLC-Q-TOF-MS), combined with chemometrics, was used to investigate the changes of compounds in Schizonepetae Herba (SH) before and after being charred. A total of 104 compounds were tentatively identified in SH and 83 in SHC. Fifteen differential compounds were found between by chemometrics SH and SHC. Altogether, our findings can provide a practical approach to the processing technology of carbonizing by stir-frying SH.

Optimization of Extraction and Quality Evaluation of Abri Herba by Response Surface Methodology Combined with Quantitative Analysis of Multi-Components by a Single Marker.

BACKGROUND: Abri Herba (AH) is a famous medicinal and edible traditional Chinese medicine, which is usually used for liver disease. To date, few studies have been conducted on the ultrasonic extraction (UAE) process for AH and the application of quality analysis of multi-components by the single-marker (QAMS) method to evaluate the quality. OBJECTIVE: To optimize the UAE process for AH, and develop and validate the quality evaluation of AH by the QAMS method. METHODS: The UAE conditions of AH were optimized by response surface methodology with the total contents of protocatechuic acid, hydantoin, gardeniaine, vicenin-2, salvoside and isosalvoside as indicators, the ultrasonic time, methanol concentration and liquid to material ratio as parameters. The content of protocatechuic acid, hypaphorine, abrine, vicenin-2, schaftoside, and isoschaftoside in 12 batches of AH was first determined by the external standard method (ESM) using HPLC. After that, based on abrine as the internal standard, the relative correction factors (RCF) for protocatechuic acid, hypaphorine, vicenin-2, schaftoside and isoschaftoside were established, and the ESM method was used to verify the QAMS method. RESULTS: The results show that the optimal extraction process parameters for AH are an ultrasonic time of 22 min, a methanol concentration of 45%, and liquid to material ratio of 26 (mL/g). The QAMS results show that the relative correction factor has good reproducibility, and there is no significant difference between the results of the ESM method and the QAMS method for each chemical constituent, indicating that the research is feasible. CONCLUSION: The optimized extraction process of AH and the established QAMS-based quality control method are stable and can be used for the quality control of AH. HIGHLIGHTS: A response surface methodology was used to optimize the ultrasonic extraction process for AH, and a QAMS method was established for evaluating the quality of AH.

Exploring Antioxidant and α-Glucosidase Inhibitory Activities in Mulberry Leaves (Morus alba L.) across Growth Stages: A Comprehensive Metabolomic Analysis with Chemometrics.

Metabolic product accumulation exhibited variations among mulberry (Morus alba L.) leaves (MLs) at distinct growth stages, and this assessment was conducted using a combination of analytical techniques including high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-mass spectrometry (LC-MS). Multivariate analysis was applied to the data, and the findings were correlated with antioxidant activity and α-glucosidase inhibitory effects in vitro. Statistical analyses divided the 27 batches of MLs at different growth stages into three distinct groups. In vitro assays for antioxidant activity and α-glucosidase inhibition revealed that IC50 values were highest at the Y23 stage, which corresponds to the 'Frost Descends' solar term. In summary, the results of this study indicate that MLs at different growth stages throughout the year can be categorized into three primary growth stages using traditional Chinese solar terms as reference points, based on the observed variations in metabolite content.

Coordinate-transformation iteration algorithm for phase modulation of arbitrary axial light distribution.

Axial light distribution modulation is widely applied in optical tweezers, hard-brittle material cutting, multilayer laser direct writing, etc. To generate arbitrary axial light distribution, the coordinate-transformation iteration (CTI) algorithm is presented. The CTI algorithm unifies equations in low and high numerical aperture (NA) scenarios, using the same iterative algorithm to produce phase computer-generated holograms. In a low NA scenario, twin-foci, flattop, and sin2 distributions have been achieved. In high NA scenarios, multirings, multifoci, and needle-like distributions have been realized in simulation with specific polarized incident beams. Since the CTI algorithm is inherently an efficient one-dimensional phase retrieval algorithm that is not limited by NA, this method has the potential to become a well-received solution for axial light distribution modulation.

Molecular Networking, Network Pharmacology, and Molecular Docking Approaches Employed to Investigate the Changes in Ephedrae Herba before and after Honey-Processing.

Raw Ephedrae herba (REH) and honey-processed Ephedrae herba (HEH) were the different decoction pieces of Ephedrae herba (EH). Honey-processing that changes REH into HEH has been shown to relieve cough and asthma to a synergistic extent. However, the chemical markers and the synergistic mechanism of HEH need to be further studied. In this study, the ultra-high performance liquid chromatography coupled with hybrid quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) and molecular networking (MN) were used to investigate the chemical composition of REH and HEH, which led to the identification of 92 compounds. A total of 38 differential chemical markers for REH and HEH were identified using principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Network pharmacology suggests that the synergistic effect of HEH in relieving cough and asthma may be due to 31 differential chemical markers acting through 111 biological targets. Among them, four compounds and two targets probably played an important role based on the results of molecular docking. This study enriched our knowledge about the chemical composition of REH and HEH, as well as the synergistic mechanism of HEH.

Stealth dicing of 1-mm-thick glass with aberration-free axial multi-focus beams.

Laser stealth dicing can realize material separation with negligible surface damage, but severe aberrations in thick materials degrade processing quality. This Letter presents a nonlinear point-to-point transformation method combined with spherical aberration compensation to achieve aberration-free axial multi-focus beams. The focus peak intensity increases 7 times at a depth of 0.5 mm after spherical aberration compensation, and reaches 44 times at 3.5 mm. Spherical aberration compensation experiments showed that the width of the heat-affected zone remains almost unchanged at different depths inside the glass, and stealth dicing experiments for 1-mm-thick glass demonstrated that aberration-free 1-focus, 2-foci, and 3-foci stealth dicing can be successfully realized.

Comparison of Volatile Oils and Primary Metabolites of Raw and Honey-Processed Ephedrae Herba by GC-MS and Chemometrics.

BACKGROUND: Ephedrae Herba (EH) is the terrestrial stem of Ephedra sinica Stapf, E. intermedia Schrenk et C. A. Mey., or E. equisetina Bge, which has been used as a diaphoretic, antiasthmatic, and diuretic. Honey-processed EH (HEH) is a widely used traditional Chinese medicine, and has a better effect of dispersing lung qi and relieving asthma and a lower effect of dispelling cold than raw EH (REH). OBJECTIVE: To understand the differences in volatile oils and primary metabolites between REH and honey-processed HEH, and to provide a reference for elucidating the mechanism behind the effect of this processing. METHODS: A system data acquisition and mining strategy was designed to investigate the differences in volatiles and primary metabolites between the REH and HEH, based on GC-MS coupled with multivariate statistical analysis, including principal component analysis and orthogonal partial least squares discriminant analysis. RESULTS: Overall, 15 volatile oils and 14 primary metabolites were shown to be potential characteristic markers differentiating REH and HEH. CONCLUSION: The results may provide a scientific foundation for comprehensively revealing the honey-processing mechanism of EH. HIGHLIGHT: Volatile oils and primary metabolites were used to distinguish REH and HEH and elucidate the processing mechanism of EH for the first time.

Nanopore Creation in Graphene by Ion Beam Irradiation: Geometry, Quality, and Efficiency.

Ion beam irradiation is a promising approach to fabricate nanoporous graphene for various applications, including DNA sequencing, water desalination, and phase separation. Further advancement of this approach and rational design of experiments all require improved mechanistic understanding of the physical drilling process. Here, we demonstrate that, by using oblique ion beam irradiation, the nanopore family is significantly expanded to include more types of nanopores of tunable geometries. With the hopping, sweeping, and shoving mechanisms, ions sputter carbon atoms even outside the ion impact zone, leading to extended damage particularly at smaller incident angles. Moreover, with lower energies, ions may be absorbed to form complex ion-carbon structures, making the graphene warped or curly at pore edges. Considering both efficiency and quality, the optimal ion energy is identified to be 1000 eV at an incident angle of 30° with respect to the graphene sheet and 400-500 eV at higher incident angles. All of these results suggest the use of oblique ion beam and moderate energy levels to efficiently fabricate high-quality nanopores of tunable geometries in graphene for a wide range of applications.

Antibiotics pollution in Jiulong River estuary: source, distribution and bacterial resistance.

To gain insight into the antibiotic pollution in the Jiulong River estuary and the pollutant sources, we analyzed the concentration of 22 widely-used antibiotics in water samples collected from the river and estuary, 17 and 18 sampling sites, respectively. Contamination with sulfonamides, quinolones and chloramphenicols was frequently detected and the distribution pattern of antibiotics suggested that most of the pollutants are from the Jiulong River, especially from the downstream watersheds. To reveal the ecological effects, we isolated 35 bacterial strains from the estuary and analyzed their antibiotic resistance to the eight most frequently detected antibiotics. The bacteria were subsequently classified into seven different genera by 16SrDNA sequencing. Up to 97.1% of the bacteria showed resistance and 70.6% of strains showed multi-resistance to these antibiotics, especially to sulfonamides. This study demonstrated a pattern of antibiotic contamination in the Jiulong River and its estuary and illustrated high bacterial antibiotic resistance which was significantly correlated with the average antibiotics concentrations and detected frequencies in the estuary.