As one crucial resource of Chinese medicines, fungal Chinese medicine have been extensively used in clinic. This article has compiled a total of 442 standards for fungal Chinese medicine that have been issued in China, including 19 Chinese Pharmacopoeia standards, 398 local standards and 25 bureau or ministry standards. The fungal Chinese medicine are composed by 67 traditional Chinese medicine varieties from 66 fungal species. Based on the review of those quality standards, the characteristics and quality evaluation of fungal Chinese medicine such as medicinal property, microscope appearance, DNA based identification and physicochemical identification, assay (polysaccharides, nucleosides, sterols, terpenes, proteins/peptides, esters, sugar alcohols, etc.), microbial and harmful substances, as well as related analytical techniques are summarized. This review will certainly provide the comprehensive understanding of current research and development of fungal Chinese medicinal materials.

Objective: To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 16 saponins (notoginsenoside R₁, notoginsenoside Fa, notoginsenoside Fe, ginsenoside Re, ginsenoside Rg₁, ginsenoside Rf, ginsenoside F₃, ginsenoside Rg₂, ginsenoside Rb₁, ginsenoside F₁, ginsenoside Rb₂, ginsenoside Rb₃, ginsenoside Rd, ginsenoside F₂, ginsenoside Rg₅, and 20(S)-ginsenoside Rg₃) in compound Sanqi capsules, and to evaluate the quality of these compound Sanqi capsules. Methods: The samples were extracted with methanol using ultrasonic extraction, and separation was performed on a Thermo Fisher Scientific Accucore Phenyl Hexyl column (2.1 mm×100 mm, 2.6 µm) using gradient elution, with 0.1% formic acid aqueous solution (A) and acetonitrile (B) as mobile phase. The flow rate was 0.4 mL · min^-1, the column temperature was 35 ℃, and the injection volume was 2 μL. Mass spectrometry was performed by using a heated electrospray ion source (HESI), and parallel reaction monitoring (PRM) in negative ion mode was employed for data acquisition and determination. The spray voltage was 2.8 kV(-). Results: The established method showed a good linear relationship within a certain concentration range (r≥0.997 0). The precision, repeatability and stability of the tested samples were good with recoveries ranging from 96.6% to 102.3%, and the RSDs between 1.1% and 5.1%. The results indicated that the contents of the 16 saponins were different among samples from different manufacturers and different batches of samples from the same manufacturer. The average total saponin contents of the samples from the three manufacturers were 33 019.650 8, 32 801.840 8, and 27 108.822 0 μg · g^-1 respectively. Among the 10 batches of samples, the one with the lowest content was ginsenoside Rf and the top five saponin components in terms of content were ginsenoside Rg₁, ginsenoside Rb₁, notoginsenoside R₁, ginsenoside Rd, and ginsenoside Re in sequence. Moreover, the average total contents of these five saponins all accounted for more than 95% of the total saponins, and could be used as markers that contribute significantly to quality differences. Additionally, cluster analysis could be divided into 3 categories according to manufacturers. Statistics showed that in the samples classified by cluster analysis as class Ⅰ, the dispersion degrees of the other saponins in the box diagram were greater than thoes in class Ⅱ and class Ⅲ. except ginsenoside Rb₂ and contents of ginsenoside Rg₅ were significantly higher than those in class Ⅱ and class Ⅲ, with a significant difference (P<0.001). The qualities of 3 batches of samples in class Ⅱ were stable, and contents of ginsenoside Rb₂ were significantly different from those in class Ⅰ and class Ⅲ (P<0.001). The components of saponins in class Ⅲ samples were lower than those in class Ⅰ and class Ⅱ samples. Conclusion: The method can quickly, efficiently and accurately determine the contents of 16 saponins in compound Sanqi capsules, and provides a reference for the quality control of compound Sanqi capsules.

Pharmacological studies have demonstrated the important role of flavonoids in promoting human health. The determination of pharmacological activity requires using of monomeric compounds. However, the advancement of precision medicine has imposed more stringent requirements on compounds, especially compounds intended for therapeutic use. In chiral drug research, the impact of optical isomers on drug efficacy should not be overlooked. While most drugs are used in clinical practice as racemates, certain isomers may not only reduce therapeutic efficacy, but increase metabolic load or even cause toxicity. Chiral stationary phases (CSP) are important materials for obtaining optically pure flavonoid compounds and represent a core aspect of chiral research. This review summarizes the recent developments in chiral stationary phases for the enantioseparation of flavonoids, in order to provide a reference for the chiral separation of flavonoids and offer new perspectives for further exploration and utilization of flavonoid compounds.

Multi-angle static light scattering (MALS) is a critical characterization technique in the field of biological macromolecules, providing comprehensive structural information such as molecular weight, molecular weight distribution, root mean square radius, second dimensional coefficient, molecular chain conformation, etc. However, the complexity of its underlying principles and multiple influencing factors often challenge the accuracy of its measurements. This review focuses on the basic theory of multi-angle static light scattering in dilute solutions of biological macromolecules and the analysis of its influencing factors in the characterization of dilute solutions of biological macromolecules including dn/dc value, second dimensional coefficient (A2), interdetector delay volume, extrapolation models, and fluorescence interference. By addressing these variables, this work aims to assist analysts in better understanding and mastering multi-angle static light scattering technology to obtain reliable and precise characterization data.

Objective: To identify different sources of Pinellia ternata (Thunb.) Breit. five-leaf germplasm resources using traditional four major identifications, DNA barcode molecular identification, and ploidy identification. Methods: 13 germplasm resources were collected and propagated in vitro through tissue culture. The origin, traits, and microscopic characteristics of the 13 resources were identified according to the Chinese Pharmacopoeia (2020 Edition, Volume Ⅰ). Thin-layer chromatography (TLC) was conducted using chloroform-methanol (6 ∶ 1) as the developing agent and 10% sulfuric acid ethanol as the color reagent for identification. ITS2 nucleic acid sequences of the resources were amplified by PCR, and sequence alignment and phylogenetic tree construction were performed using Snapgene and Mega software. Chromosome counting was used to identify the ploidy of the Pinellia ternata five-leaf. Results: All five-leaf resources were consistent with the description of Pinellia ternata in the “Flora of China,” and were preliminarily identified as Pinellia ternata. The traits and microscopic characteristics of the five-leaf also aligned the description of Pinellia ternata in the “Chinese Pharmacopoeia” (2020 Edition, Volume Ⅰ). TLC analysis showed that the traditional Pinellia ternata and the five-leaf had the same chromatographic profiles and exhibited clear differentiation from Rhizoma Arisaematis (tiger palm). The amplification and sequencing success rate of ITS2 was 100%. Phylogenetic analysis based on the ITS2 sequences in the NCBI database revealed that the five-leaf was closely related to Pinellia ternata and clustered together with traditional Pinellia ternata resources. Chromosome counting indicated that three germplasm resources from Yunnan (SY-3, WY-8, and WY-9) were hexaploid (2n=6x=78), while nine germplasm resources from Hubei (SY-1, SY-2, WY-1, WY-2, WY-3, WY-4,WY-5, WY-6, and WY-7) were octaploid (2n=8x=104). One tiger palm resource was diploid (2n=2x=26). Conclusion: The five-leaf is identified as a Pinellia ternata resource,and the determination of its ploidy levels provides an important basis for developing new germplasm.

Objective: To establish a standardized method for the in vitro bioactivity analysis of polyethylene glycol-conjugated recombinant human growth hormone (PEG-rhGH) based on Nb2-11 cell proliferation and to study the feasibility of replacing in vivo animal bioassay with the Nb2-11 cell bioassay. Methods: The in vitro biological activity of 9 batches of PEG-rhGH injection was detected by the Nb2-11 cell method to draft the effective experimental standard. The method was validated in accordance with the General Chapter 9401 of the 2020 Edition of the Chinese Pharmacopoeia. Four laboratories collaborated to detect the biological activity of 2 batches of PEG-rhGH drug substance and 9 batches of injection to study the precision of the method. The biological activity of 8 batches of PEG-rhGH drug substance and 9 batches of PEG-rhGH injection was detected by the Nb2-11 ell method and in vivo animal method respectively to study the consistency of the results determined by in vitro and in vivo methods. The in vitro biological activity of 24 batches of PEG-rhGH injection with different expiration dates was detected to study the standard limit of the in vitro biological activity of PEG-rhGH injection. Results: The relative accuracy of the method showed a relative bias ranging from -3.059% to 3.557%. The linear regression slope was 0.984, with intermediate precision geometric coefficients of variation (GCV) of 5.667%-6.923%. The pass rate for the proposed effective experimental standards was 100%, the inter-laboratory geometric coefficient of variation was 2.281%, and the average ratio of in vivo and in vitro test results was 0.938. The relative potency of PEG-rhGH products with different expiration dates ranged from 100% to 119%. Conclusion: This method exhibits good reproducibility both within and between laboratories and showed good consistency with the in vivo animal bioassay. It can serve as a standardized alternative to animal testing for quality control and release testing of PEG-rhGH products.

Objective: To establish the fingerprint of Qingbai Tongbi capsules, and to screen out its indicative compounds for quality control combined with chemometrics methods and network pharmacology. Methods: Agilent 5 TC-C18(250 mm×4.6 mm, 5 μm) chromatographic column was used for separation. The mobile phase was acetonitrile-0.1% phosphoric acid solution for gradient elution at a flow rate of 1.0 mL · min-1, and the column temperature was 30 ℃. The injection volume was 10 μL, and the wavelength was 210 nm (0-45 min), 260 nm (45-70 min). The fingerprint was established and the common peaks were determined. By comparing with the relative retention time and UV spectra of the reference substances, the corresponding compounds of the chromatographic peak were identified. Then the common peaks were identified by single decoction. Chemometrics methods was used to evaluate quality of 10 batches of Qingbai Tongbi capsules, and OPLS-DA analysis was used to screen out the main marker components of Qingbai Tongbi capsules. Combined with network pharmacology, the core targets and key pathways were constructed a “component-target-pathway” network map through corresponding databases. Combined with the above results, indicative compounds for quality control of Qingbai Tongbi capsules were screened out, and an HPLC method was established to determine the content of the Q-Markers. Results: An HPLC fingerprint of Qingbai Tongbi capsules was established, identifying 24 common peaks, and assigning them to different peaks. Among them, peaks 1-3, 5-8, 10-11, 13 and 16 came from Qingfengteng. Peaks 4, 9, 12, 14, 16, 18-19 came from Baishao. Peaks 15, 17, 20-24 came from Zhigancao. Eight common peaks were identified, including catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose, glycyrrhizic acid. The similarity evaluation showed that the similarity of 10 batches of Qingbai Tongbi capsules samples ranged from 0.934-1.000. Principal component analysis (PCA) showed that the cumulative variance contribution rate of the first four principal components was 93.998%, while orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that 8 components had higher variable importance projection values. On this basis, the network pharmacology method was used to analyze and conclude that catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose and glycyrrhizic acid may be the potential Q-Marker of Qingbai Tongbi capsules. The contents of the above eight components were determined simultaneously, and the methodological investigation results were good. The average sample recovery rate was 96.81%-103.44%, and the RSD was 0.6%-3.7%. The mass fractions of catechin, sinomenine, gallic acid, magnoflorine, paeoniflorin, glycyrrhizin, 1, 2, 3, 4, 6-O-pentagalloylglucose and glycyrrhizic acid in 10 batches of samples were 0.907 1-1.189 3 mg · g-1, 2.183 3-3.118 6 mg · g-1, 0.397 0-1.427 6 mg · g-1, 3.507 9-5.446 6 mg · g-1,14.207 7-19.570 1 mg · g-1, 1.412 8-3.577 5 mg · g-1, 0.442 0-1.697 7 mg · g-1, 2.738 8-4.761 2 mg · g-1. Conclusion: The established HPLC fingerprint method is simple and good repeatability. The quality control indicative compounds of Qingbai Tongbi capsules can provide a basis for its quality control.

Objective: To establish the HPLC fingerprint and evaluate the quality of different batches of Folium Cinnamoni by chemometrics and quantitative analysis of multi-components with a single-marker (QAMS). Methods: HPLC was used to establish the fingerprints of 12 batches of Folium Cinnamoni and perform similarity evaluation. And then principal component analysis (PCA) and cluster analysis (CA) were used to analyze the chemical pattern of different batches of Folium Cinnamoni. The QAMS method was established by using cinnamic acid as the internal standard reference. The relative correction factors of 2-hydroxy cinnamaldehyde, cinnamyl alcohol, trans-cinnamaldehyde and 2'-methoxy cinnamaldehyde were calculated. The contents of 2-hydroxycinnamaldehyde, cinnamyl alcohol, cinnamic acid, trans-cinnamaldehyde and 2'-methoxy cinnamaldehyde in Folium Cinnamoni were determined by external standard (ESM) method and QAMS method. Results: A total of 12 common peaks were identified from the fingerprint, and 7 of them were confirmed by reference substance. CA divided 12 batches of Folium Cinnamoni into 2 categories; PCA showed that there were some differences in the chemical composition of them. Three principal components were identified, and the result showed that the cumulative variance contribution rate was 86.022%. The contents of these 5 compounds measured by QAMS and ESM showed no significant difference. Conclusion: The fingerprint and content determination method of Folium Cinnamoni are stable and reliable, which can provide reference for quality control and evaluation of Folium Cinnamoni.

Objective: To comprehensively characterize the component profile of polysorbate 80 (PS80) using ultra high performance liquid chromatography-quadrupole time-of-flight/mass spectrometry (UHPLC-Q TOF/MS), to identify its degradation products, to investigate the effects of different buffer systems (citrate, phosphate, and histidine buffers), acid-base hydrolysis, and oxidative degradation on the PS80 component profile, to identify new degradation products of PS80 and its degradation behavior and potential mechanisms under various stress conditions. Methods: PS80 and its stress samples (subjected to acid-base hydrolysis or Fe²⁺-induced oxidative degradation in citrate, phosphate, and histidine buffer systems) were separated using an Agilent ZORBAX RRHD SB-C₈ column (100 mm×2.1 mm, 1.8 μm) with gradient elution employing 0.1% formic acid in water and 0.1% formic acid in acetonitrile as the mobile phase. Mass spectrometric analysis was performed in positive electrospray ionization (ESI⁺) mode with full-scan MS (m/z 100-3 000) and MS/MS under collision energies of 80 eV (for doubly charged ions) and 120 eV (for singly charged ions). Molecular ions, fragment ions, and degradation patterns of PS80 components were analyzed to deduce the structures of degradation products and elucidate potential degradation pathways. Results: Nine major components of PS80 were successfully separated and identified using UHPLC-Q TOF/MS. PS80 exhibited greater stability under acidic conditions compared to alkaline conditions. In histidine buffer, Fe²⁺ readily induced oxidative degradation of unsaturated esters in PS80, generating four representative oxidative degradation products: polyoxyethylene iso-sorbitol 9-oxononanoate monoester, polyoxyethylene iso-sorbitol epoxy stearate monoester, polyoxyethylene sorbitan ketostearate monoester, and polyoxyethylene ketostearate monoester. Conclusion: The UHPLC-Q TOF/MS technique is effectively utilized to comprehensively identify and characterize the component profile of PS80. The findings enhance the understanding of PS80 behavior in complex pharmaceutical systems and provide valuble insights for pharmaceutical formulation design.

Objective: To provide an experimental basis of the new fermentation process of multi-strain, based on the previous studies on the strain separation and processing principle of Arisaema Cum Bile, and the dominant strain is screened out and the process of multi-strain fermentation process was further optimized. Methods: Every strain isolated in the previous stage was single fermented to prepare a series of Arisaema Cum Bile. The content of hyocholic acid, hyodeoxycholic acid, and chenodeoxycholic acid was determined using HPLC-ELSD and screened for the advantageous strain. Taking the total amount of free bile acid and the content of taurine, glycine, and bilirubin as the indexes, making the addition amount, fermentation temperature, and fermentation time of the multi-strains as the factors, and combining the orthogonal test and multi-index weighted scoring method, the fermentation process was investigated and the new fermentation process of the multi-strains was determined. Results: The advantageous strains were identified as Lactobacillus plantarum and Enterococcus (anaerobic). A new process of multi-strain fermentation has been screened out: mix with a ratio (1 ∶ 1) of Arisaematis Rhizoma powder and bile containing 10%-15% multi-strains liquid [1 ∶ 1 mixture of Lactobacillus plantarum and Enterococcus (anaerobic)], ferment at 32 ℃ with 80% humidity for 15 days, take out, steam for 1 h to pass thoroughly, cut into diced, and dry in the sun. Conclusion: The optimized fermentation process for Arisaema Cum Bile is stable and feasible, which can improve the quality of the products and provide scientific basis for the standardized production of Arisaema Cum Bile.

Objective: To study the compatibility stability and pharmacokinetics of nimodipine concentration for injection in Beagle dogs. Methods: A Hyper Star BDS C18 column (150 mm×4.6 mm, 5 μm) was used; the mobile phase was water-tetrahydrofuran-acetonitrile; the detection wavelength was 235 nm; the injection volume was 10 μL;the flow rate was 1.0 ml · min-1; the column temperature was 30 ℃; the running time was 30 min. A HALO 90A AQ-C18 column (3.0×30 mm, 2 µm) was used; the mobile phase A was 95% water/5% acetonitrile (0.1% formic acid), and mobile phase B was 95% acetonitrile/5% water (0.1% formic acid), with gradient elution; the flow rate was 0.6 mL · min-1; the auto-sampler temperature was 4 ℃; the running time was 2.30 min, and the injection volume was 8.00 μL. The mass spectrometry conditions were as follows: electrospray ionization source, positive ion mode, curtain gas was 35 psi, collision gas was 10 psi, spray voltage was 4500 V, nebulizer temperature was 500 ℃, nebulizer and auxiliary gas were both 50 psi. The m/z of nimodipine was 417.18→122.20, and that of dexamethasone was 437.10→361.00. The contents of nimodipine concentration for injection and Nimotop® injection after mixing with different solutions were determined using HPLC, and plasma concentrations of nimodipine following administration were measured using HPLC-MS/MS in Beagle dogs. The pharmacokinetic parameters were calculated using WinNonlin 8.3.4 software, and the main pharmacokinetic parameters were statistically analyzed using SPSS Statistics 26 software. Results: The nimodipine concentration for injection demonstrated good compatibility with the test solutions, and the content of the novel nimodipine formulation remained stable with no significant change within 48 h after compatibility. In contrast, the content of the original preparation agent decreased significantly after compatibility, accompanied by visible crystal formation. There were no significant differences in Cmax, Tmax and AUC of the novel nimodipine formulation compared with the original preparation agent (P>0.05). Conclusion: Nimodipine Concentration for injection exhibits good stability and does not change the pharmacokinetic characteristics of the API, suggesting good clinical application prospects.

Objective: To assess the risk of introducing bovine and porcine viruses in the production of human mesenchymal stem cells, which may involve the use of raw materials such as bovine serum and trypsin, this study infected human mesenchymal stem cells (HMSC) with various bovine and porcine viruses. The aim was to provide a foundation for risk assessment in the prevention and control of these viruses in HMSC. Additionally, by comparing the infection and proliferation patterns of bovine and porcine viruses in human cell lines 293T/17 and MRC-5, this study offered a comprehensive analysis of the susceptibility of HMSC to common bovine and porcine viruses. Methods: HMSC, human diploid MRC-5 cells, 293T/17, Vero, and PK-15 cells were seeded in 24-well plates and 25 cm2 flasks at a density of 7×103 cm-1, respectively, and then inoculated with porcine parvovirus (PPV) at a multiplicity of infection (MOI) of 0.02. Similarly, HMSC, MRC-5, 293T/17, Vero, and BT cells were seeded in 24-well plates and 25 cm2 flasks at a density of 7×103 cm-1, respectively, and inoculated with bovine parvovirus (BPV), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus type 3 (PI3), bovine adenovirus type 3 (BAV-3), and reovirus type 3 (REO-3) at an MOI of 0.02. The cells in the 24-well plates were cultured for 7 days, while those in the 25 cm2 flasks were blindly passaged for three generations. The third generation of blind-passaged cells were transferred to 24-well plates. For each passage, cytopathic effects (CPE) were observed, immunofluorescence assays were performed, and viral nucleic acids were detected by qPCR. Results: Compared to PPV-sensitive PK-15 cells, no CPE or fluorescence were observed in Vero, HMSC, or MRC-5 cells after PPV inoculation, and the PPV copy number continuously decreased with successive passages. In contrast, although no CPE was observed in 293T/17 cells, green fluorescence was detected after three blind passages, and the PPV copy number remained consistently high at 1.49×107 copies · mL-1. Compared to bovine virus-sensitive BT cells, HMSC also exhibited susceptibility to three bovine viruses (PI3, REO-3, and BPV), showing varying degrees of CPE and positive immunofluorescence upon infection. Viral nucleic acids were detected via qPCR for all three viruses. Among them, PI3 and REO-3 were able to replicate in HMSC, though at lower levels compared to 293T/17 cells. Conclusion: HMSC and MRC-5 cells exhibit similar viral susceptibility profiles, demonstrating permissiveness to BPV, REO-3, and PI3, but resistance to PPV, BVDV, and BAV-3. In contrast, 293T/17 cells show susceptibility to PPV, REO-3 and PI3, while remaining non-permissive to BPV, BVDV and BAV-3. In the control of viral safety risks in human mesenchymal stem cells, greater attention should be paid to the infection risks of BPV, REO-3, and PI3 viruses when introducing porcine or bovine-derived materials of animal origin.

Objective: To establish a magnetic solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (MSPE-UHPLC-MS/MS) method for rapidly detecting 14 fentanyl analogs in wastewater. Methods: Wastewater samples were purified and concentrated using a novel mixed-mode weak cation exchange magnetic solid phase extraction adsorbent, Fe3O4@poly (ST/DVB/MA-COOH), followed by analysis with UHPLC-MS/MS method. Separation was performed on a Waters Acquity UPLC HSS T3 column (150 mm×2.1 mm, 1.8 μm)at 40 ℃. The mobile phase consisted of 0.1% formic acid-aqueous solution(A) and 0.1% formic acid-acetonitrile solution(B) with gradient elution. The flow rate was 0.3 mL · min-1, and an injection volume was 2.5 µL. Detection was carried out using electrospray ionization in multiple reaction monitoring mode. Results: The calibration curves for the 14 fentanyl analogs showed good linearity over the range of 0.04-4 ng · L-1 (r≥0.998 4). The limit of detection (LOD) was 0.013 ng · L-1 and the limit of quantitation (LOQ) was 0.04 ng · L-1. The recovery rates at low, medium and high spiked levels ranged from 85.4% to 114.7%. The intra-day precision(n=6) was between 2.0% and 6.4%, and the inter-day precision(n=3) was between 0.4% and 2.4%. Fentanyl was detected in five actual wastewater samples, with concentrations ranging from 0.21 to 0.46 ng · L-1. Conclusion: This method has the advantages of short pretreatment time, simple operation, low solvent consumption, high sensitivity, and good reproducibility. It is suitable for high-throughput screening and quantitative analysis of fentanyl analogs in wastewater samples.

Objective: To identify the impact of microbial contamination from environments on quality of pharmaceuticals, pertinent strategies were developed and information for reference was provided. Methods: Systematic collection, collation and analysis of microbial characteristics and risk profiles from authoritative sources (including Bergey's Manual of Systematic Bacteriology, peer-reviewed literature and regulatory agency guidelines) were conducted. A standardized annotation framework tailored to pharmaceutical risk assessment of microbial contamination was developed, with structured Critical microbial attributes integrated into a MySQL-based knowledge base management system. Meanwhile, a user-friendly front-end interface was developed using Vue.js and Node.js, with interactive data visualization implemented via ECharts enabling rapid querying and statistical analysis. Results: A cloud-based query platform, DM-Mpedia (http://dmcloud.dmicrobe.cn/#/preview), was established, encompassing 20 678 microbial species with detailed taxonomic, phenotypic and risk metadata. Conclusion: The platform serves as a powerful tool for microbiologists to assess and control microbial contamination or objectionable microorganisms more effectively.

Objective: To investigate the microbial contamination and community characteristics of Chinese herbal pieces (CHP) of “Zhebawei”, and provide the basis for the revision of microbial limit standards. Methods: According to the 2020 edition of the Chinese Pharmacopoeia, the total aerobic microbial count (TAMC), total combined yeasts and molds count (TYMC), total heat-resistant microbial count (HRMC), and specified microorganisms were detected in 40 batches of samples, and the community characteristics of contaminated microorganisms in CHP of “Zhebawei” were explored based on 16S rDNA high-throughput sequencing. Results: The lgTAMC ranged from 0.50 to 7.87, and the lgTYMC ranged from 0.50 to 6.72 in 40 batches of samples. The detection rates for HRMC and bile-tolerant Gram-negative bacteria (BTGB) were 72.5% and 75% respectively, and the BTGB in 3 batches of samples exceeded 104 cfu · g-1. Escherichia coli and Salmonella were not detected in any samples, but important opportunistic pathogens such as Cronobacter sakazakii, Klebsiella pneumoniae and Enterobacter aerogenes were identified. The results of 16S rDNA high-throughput sequencing showed that the contaminated microorganisms in CHP of “Zhebawei” were distributed in 25 phyla and 538 genera. There were significant differences in the dominant bacteria genera among different varieties of CHP, and the genera containing common pathogenic bacteria or opportunistic pathogenic bacteria such as Acinetobacter, Serratia, Flavobacterium and Escherichia-Shigella were detected. Conclusion: More comprehensive information of contaminated microbial communities can be obtained from 16S rDNA high-throughput sequencing. The degree and characteristics of microbial contamination in different varieties of CHP is significantly different. Various pathogenic bacteria are identified indicate that manufacturers and distributors of CHP should prevent and control the risks of microbial contamination. It is also advised the relevant departments to improve the evaluation standard of microbial limit according to the classification of CHP.

Dalbergiphenols are a group of characteristic components in plants of Dalbergia, possessing biological activities such as anti-osteoporosis, cardioprotective, antibacterial, antioxidant, and anti-inflammatory properties. This paper analyzes the data from ¹H nuclear magnetic resonance (¹H-NMR) and ¹³C nuclear magnetic resonance (¹³C-NMR) spectra of 19 dalbergiphenols to summarize the patterns of variations between the corresponding chemical shifts and their substituents. This review aims to provide information for the rapid identification of new dalbergiphenols in the future.

Objective: To establish an HPLC method for the simultaneous determination of several key constituents, namely syringin, echinacoside, verbascoside, epimedin A, epimedin B, epimedin C and icariin in Cistanches Herba formula, aiming to provide experimental evidence for the quality control of the formula and serve as a material basis for research on its anti-fatigue efficacy. Methods: The analysis was performed on a ZORBAX Eclipse Plus C₁₈ column (250 mm×4.6 mm, 5 μm) at 25 ℃, with the mobile phase comprising of acetonitrile-water flowing at 1.0 mL · min^-1 in a gradient elution manner. The detection wavelengths were set at 210 nm, and the injection volume was 10 μL. Results: The content determination method for the signature constituents in the Cistanches Herba formula was established. The linear relationship of the seven constituents was excellent within their respective ranges (r≥0.999 7), and the RSD values of precision, repeatability, and stability were all below 5%. The average recovery rates ranged from 93.5% to 104.0%, with RSD values between 0.11% and 2.5%. The content ranges of the seven constituents in four batches of samples were as follows: syringin (0.12-0.18 mg · g^-1), echinacoside (16.99-18.65 mg · g^-1), verbascoside (1.35-1.76 mg · g^-1), epimedin A (0.18-0.21 mg · g^-1), epimedin B (0.83-0.98 mg · g^-1), epimedin C (0.82-0.98 mg · g^-1), and icariin (0.58-0.83 mg · g^-1). Conclusion: This method demonstrates good sensitivity, strong specificity, and excellent repeatability. It can serve as a reliable basis for the quality evaluation and control of Cistanches Herba formula.

Objective: To investigate the effect of alisol G (AG) on lipid metabolism during 3T3-L1 preadipocyte differentiation. Methods: The differentiation of 3T3-L1 preadipocytes was induced by a combination of insulin, 3-isobutyl-1-methylxanthine, and dexamethasone. AG was added to mediate the process. Oil red O staining method was performed after intervention with AG. The contents of the relative lipid, triglyceride (TG) and total cholesterol (TC) within the cells were determined. Ultra performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry (UPLC-Q TOF-MS/MS) techniques were adopted to analyze changes in the lipid profile and the impact of lipid metabolism pathways. Results: The contents of lipid, TG, and TC during the differentiation process of 3T3-L1 preadipocytes were significantly reduced by 5 mol · L^-1 and 10 mol · L^-1 AG (P<0.01). After cellular lipidomics analysis, potential differential lipid metabolites were mainly fatty acid (FA) and glycerophosphatide (GP). KEGG enrichment analysis showed that AG affected the differentiation of 3T3-L1 preadipocytes mainly through Biosynthesis of unsaturated fatty acids, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, glycerophospholipid metabolism and other metabolic pathways. Conclusion: AG inhibited 3T3-L1 preadipocyte differentiation to a certain extent, suggesting its potential to ameliorate lipid metabolism disorder. The results of lipidomics showed that AG could improve the abnormal state of lipid metabolism during adipocyte differentiation by regulating the levels of fatty acid and phospholipid metabolites.

Objective: To compare the moxa yield and chemical components in different botanical parts of Artemisia stolonifera. Methods: The moxa yield of leaves on the middle stem (L), young leaves on the lateral branch (SL), inflorescences (F), non-lignified stems (S) and lignified stems (OS) of A. stolonifera were determined,and the chemical components were identified and analyzed by gas chromatography-quadrupole-time-of-flight tandem mass spectrometry (GC-Q TOF MS/MS). Principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and variable importance in projection (VIP) were used to find the characteristic components of A. stolonifera. Results: The highest moxa yield was obtained from the young leaves on lateral branches of A. stolonifera. A total of 19 compounds were detected, with terpenoids being the predominant class. Among the five botanical parts, six common volatile components were identified, which were β-caryophyllene, humulene, germacrene D, bicyclogermacrene, spathulenol, and caryophyllene oxide. Cluster heatmap revealed that L and S groups clustered together, whereas the F, SL, and OS formed another cluster. PLS-DA analysis pinpointed 6 components that significantly contributed to the differentiation among various botanical parts of A. stolonifera. Conclusion: Variations in the moxa yield, chemical constituents, and the contents of volatile components are observed among different botanical parts, which provides theoretical and data references for the further development and application of A. stolonifera.