To establish the ultra performance liquid chromatography (UPLC) characterization profile of Styrax, and to determine the contents of three components (cinnamic acid, cinnamyl cinnamate, 3-phenylpropyl cinnamate) in Styrax, the aim was to study the quality of commercially available sulforaphane of different origins and forms, and to provide a reference for the quality control of imported medicinal herbs Styrax.

The UPLC method was performed on an Acquity UPLC BEH C₁₈ (100 mm×2.1 mm, 1.7 µm) column with acetonitrile-1% acetic acid solution as the mobile phase in a gradient elution. The volume flow rate was 0.3 mL·min^-1. The detection wavelength was 277 nm. and the temperature of the column was 35 ℃. Characteristic chromatograms of styrax were established, and the contents of three major components of Styrax from different sources were determined and the data were statistically analyzed by hierarchical cluster analysis (HCA) and principal component analysis (PCA) with ChemPattern software.

The results showed that the UPLC characteristic chromatograms of Styrax was established with 10 common peaks and 4 identified components. The linear relationship of 3 components in their respective ranges was good (r≥0. 999), and the average recoveries (n=6) were 101.8% (RSD=1.3%), 105.8% (RSD=1.2%), 99.2% (RSD=1.8%), respectively. The content of 3 components were 2.74%-3.69%, 28.21%-30.63%, 16.89%-20.98%, respectively.

The method is simple, accurate and reproducible, and can provide an overall quality control basis for the quality standard of Styrax. Combined with the information of origin research, it has important reference significance for the expansion of the origin of Styrax, as well as the harmonization and improvement of the standards of Styrax in different countries.

To study the bioactive components of the hypoglycemic effect of the traditional Chinese medicine Polygoni Cuspidate Rhizoma et Radix through its ability to inhibit α-glucosidase activity.

Four modules: high-performance liquid chromatography system, the nanofraction collector, the channel of bioassay and high-resolution mass spectrometry were integrated to build a high-throughput bioassay profiling analysis platform. The preparative liquid chromatography was introduced to apply segmented enrichment to the extract of Polygoni Cuspidate Rhizoma et Radix, and the rapid screening of α-glucosidase inhibitors from Polygoni Cuspidate Rhizoma et Radix was realized by optimizing the chromatographic separation conditions, microfluidic fractionation collection parameters and bioassaying of enzyme activity in multi-well plates.

28 α-glucosidase inhibitors were identified from Polygoni Cuspidate Rhizoma et Radix. The inhibitory activities of catechin-3-O-galloyl and procyanidin B₂-3''-O-gallate were determined with the IC₅₀ values of (9.49±1.93) μmol·L^-1 and (69.94±8.14)μmol·L^-1, respectively.

This study has enabled high-throughput and high-resolution screening of α-glucosidase inhibitors in traditional Chinese medicine, providing a valuable tool for elucidating the active components and mechanisms of Polygoni Cuspidate Rhizoma et Radix to lower hyperglycemia.

To establish a new method for in situ visualization of spatial distribution characteristics of various secondary metabolites in the root of Wendang (Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen), so as to realize the tissular localization of secondary metabolites, and to provide a reference for the in-depth excavation of the Wendang.

Matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) was used for the mass spectrometry imaging analysis of the root metabolites in Wendang. The matrix was DHB⁺/DHB^- (30 mg·mL^-1), the substrate flow rate was 20 μL·min^-1, the nitrogen flow rate was 5 L·min^-1, the nozzle movement speed was 3 mm·s^-1, the nozzle temperature was 60 ℃, and the spraying time of each slice was 50 min. The laser energy intensity in positive ion detection mode was 60%, and that in negative ion detection mode was 40%. The detection mass range was m/z 70-1 050, the mass resolution was 70 000, the spatial resolution was 50 μm, and the pixel size was 420 px×200 px. At the same time, the pathway enrichment analysis was also carried out for the metabolites.

A total of 214 metabolites were detected, and the spatial distribution characteristics of 40 representative metabolites were visualized. The distribution patterns of different kinds of secondary metabolites in the cork - phloem - xylem varied considerably, with flavonoids mainly distributed in xylem, alkaloids, phenolics, and carboxylic acids mainly in the cork and phloem, phenylpropanoids and quinones mainly in the cork, amino acids were abundant in the phloem, and nucleotides were distributed throughout the root tissues, the indicator components atractylenolide Ⅲ and syringoside were distributed in the cork, and lobetyolin was distributed in both the cork and the xylem. Pathway enrichment analysis showed that metabolites were significantly enriched in metabolic pathways, biosynthesis of secondary metabolites, flavonoid biosynthesis, biosynthesis of amino acids, and carbon metabolism pathways.

This study visualizes the spatial distribution characteristics of metabolites in the roots of Wendang. The results of this study can provide certain theoretical support for the quality control, the identification, the extraction and separation of active ingredients, and the metabolic pattern of Wendang.

To establish a atomic absorption spectroscopy (AAS) method for detecting calcium content in recombinant human coagulation factor Ⅷ for injection, and evaluate its uncertainty.

A calcium hollow cathode lamp light source was used. Air acetylene flame atomizer. Detection wavelength 422.7 nm. 8.9% lanthanum oxide solution was used as an ionization inhibitor. Conduct inspections on specificity, accuracy, precision, quantification limit, linearity and range, stability, etc. in accordance with the validation guidelines for drug quality standard analysis methods in Part 9101 of the 2020 edition of the Chinese Pharmacopoeia. The established method was applyed to detect the content of 6 batches of samples.

There was a good linear relationship between calcium element and absorbance in the concentration range of 0.0-5.0 μg·mL^-1, with r=0.999 8. The detection line was 0.071 6 μg·mL^-1; The quantification limit was 0.216 9 μg·mL^-1. The recovery rate of the spiked sample was 100.8% to 103. 2%. The repeatability RSD was 0.64%, and the solution was stable within 8 hours. The content determination results of 6 batches of samples all complied with the regulations. The uncertainty assessment result was (117.12±4.82) μg·mL^-1, k=2.

AAS method is established for the detection of calcium content in recombinant human coagulation factor Ⅷ for injection. This method has the advantages of high sensitivity, fast analysis speed, good repeatability, high accuracy, and cost savings. At the same time, the uncertainty of this method is evaluated, which can provide quantitative evaluation indicators for the quality control of this variety.

To establish a method to determination of the related substances and polymer impurities in cefminox sodium for injection.

Cefminox sodium was degraded in high temperature to prepare degradation solution. An RP-HPLC method for the related substances analysis was established with a Kromisil C₁₈ column (250 mm×4.6 mm, 5 μm), using 10 mmol·L^-1 phosphate buffer solution (pH 2.0)-acetonitrile (98：2) (A)-acetonitrile (B) with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was maintained at 25 ℃, the detection wavelength was set at 254 nm, and the injection volume was 20 μL. The specificity of RP-HPLC method and identification of unknown impurities was researched by 2D HPLC-MS/MS.

14 main impurities were characterized in the degradation solution, including 3 cefminox dimmers and isomers which were characterized firstly. The impurities were determined in 7 batches of samples by principal component self-control with correction factor, the contents of impurity 3 were 0.01%-0.13%, the contents of impurity 4 were 0.10%-0.16%, the contents of impurity 5 were 0.02%-0.07%, the contents of impurity 6 were 0.04%-0.07%, the contents of polymer impurities were 0.03%-0.06%, the maximum single impurity contents were 0.01%-0.03%, while the total impurity contents were 0.28%-0.63%.

Cefminox degradation solution in high temperature can be used to identify related impurities and polymer peaks as the systematic suitability testing solution. The RP-HPLC method was suitable for related substances as well as polymer impurities in cefminox sodium for injection. This work provides useful information for the quality control of cefminox sodium, which can contribute to establishment of reasonable impurity limits.

To optimize an LC-Q TOF MS method for the determination of target proteins expressed in recombinant cells from a cell bank.

The recombinant Chinese hamster ovary (CHO) cell line expressing antibodies was selected. The supernatant of recombinant CHO cell suspensions was retained after 12 500 r·min^-1 centrifuging for 10 min and treated with 50 mmol·L^-1 NH₄HCO₃ solution. Chymotrypsin was added for enzymatic digestion, and the comparative analyses were carried out using two LC-Q TOF MS with Advance Bio Peptide(100 mm×2.1 mm, 2.7 μm) column, the Sciex TRIPLE TOF 5600+ and the Agilent Q TOF G6545, respectively. To establish the database, the amino acid sequences of the characteristic peptides of the protein heavy and light chains were entered into SCIEX BioPharmaView Version 3.0 and Agilent MassHunter BioConfirm 12.0 software, respectively. The peptides detected in the samples were searched to identify the target peptides.

The samples were analysed by a Sciex TRIPLE TOF 5600+ instrument to obtain 8 characteristic peptides with 100%amino acid sequence coverage and the samples were analysed by an Agilent Q TOF G6545 instrument to obtain 12 characteristic peptides with 100% amino acid sequence coverage.

The method established in this study is simple to operate, has high coverage and good stability, can be used to identify the expression of target proteins in recombinant CHO cell lines. It also provides a reference for the expression identification of target proteins in similar cell lines.

To establish an HPLC method for simultaneous determination of vancomycin, caffeine and aminophylline in serum of children patients and apply it to clinical detection.

6% perchloric acid was used as protein precipitator and acetanilide was used as internal standard. The determination was performed on Nucifera C₁₈ column (250 mm×4.6 mm, 5 μm) with mobile phase methanol-0.05 mol·L^-1 potassium dihydrogen phosphate (18 ：82, v/v). The detection wavelength was 236 nm, the flow rate was 1 mL·min^-1, the column temperature was 25 ℃, and the sample size was 20 μL.

The serum concentrations of vancomycin and aminophylline were in the range of 1.0-100 μg·mL^-1, and the standard curves were Y=0.010 6X+0.006 9(r=0.998 8), Y=0.012 7X+0.008 4 (r=0.998 9), respectively. The serum concentration of caffeine had a good linear relationship in the range of 1.0-160 μg·mL^-1, and the standard curve was Y=0.015 4X-0.010 1 (r=0.999 1). Quantitative downlines were all 1.0 μg·mL^-1. The average recovery rates of the three compounds ranged from 97.8%to 103.1%, and the relative standard deviations of intraday and daytime precision were less than 10%, indicating good sample stability. Ten cases of children receiving vancomycin, aminophylline or caffeine were collected for method verification, which met the needs of clinical detection.

The method is simple, rapid, accurate, and is suitable for the simultaneous determination of serum vancomycin, caffeine and aminophylline in children.

To identify the structure and determine the source of unknown impurities present at levels greater than 0.1% in representative pharmaceutical excipient benzalkonium chloride samples, in accordance with ICH Q3A guidelines.

A new LC-MS method suitable for the structure prediction of related substances of benzalkonium chloride was established. The separation was performed on an Acclaim 120 C₁₈ column (250 mm×4.6 mm, 5 μm) using a mobile phase consisting of methanol (mobile phase A) and 20 mmol·L^-1 ammonium formate aqueous solution (pH 3.5, mobile phase B) with gradient elution at a flow rate was 1 mL·min^-1. The post-column split was set to 1 ：3. MS data were collected in positive ion mode using an electrospray ionization (ESI) source. The structures of unknown impurities were inferred using a “Diagnostic fragment ion extension strategy” and confirmed by comparing the chromatographic and mass spectrometric behaviors of the impurities with those of reference substances. Additionally, the source attribution and genotoxicity prediction of the detected impurities were performed.

A total of five unknown impurities were detected in a representative sample of benzalkonium chloride by the newly established LC-MS method, and their structures were inferred. The structures of four impurities were confirmed using two commercially available reference substances and two reference substances synthesized in a directionally oriented manner. Impurity D was identified as N-benzyl-N,N-dimethyl-1-phenylmethanaminium chloride. Impurity E as N-benzyl-N-methyl-1-phenylmethanamine. The impurity G as N-benzyl-N-methyldodecan-1-amine, and impurity H as N,N-dibenzyl-N-methyldodecane-1-ammonium chloride. Impurity F was hypothesized to be an isomer of the formula C₁₅H₁₇N. Predictions using Nexus 2.6.0 software indicated that all the above impurities fell into category 5 and had no genotoxic potential. Furthermore, the method successfully located four unknown impurities detected by the USP method in benzalkonium chloride related substances using impurity reference standards.

This study systematically examines the structure and safety risks of potential impurities in benzalkonium chloride, providing valuable insights for enhancing quality control standards and pharmacopoeia criteria both domestically and internationally.

To confirm the validity of solvent/detergent (S/D) treatment and dry heating for inactivation of virus in C1 esterase inhibitor (C1-INH).

The Sindbis virus in samples with S/D was inactivated by the S/D method, and the virus titer before and after inactivation was detected using the plaque assay. Dry heating method was used to inactivate encephalomyocarditis virus (EMCV) and porcine parvovirus (PPV), and cytopathic assay was used to detect the virus titer before and after inactivation.

After inactivation by the S/D method, the reductions of Sindbis virus in three batches of smaples with S/D were > 4.35 lgPFU·mL^-1, > 4.51 lgPFU·mL^-1,> 4.64 lgPFU·mL^-1, respectively. After dry heating, the reductions of EMCV in three batches of samples without S/D were ≥5.38 lgTCID₅₀/0.1 mL、≥5.12 lgTCID₅₀/0.1 mL、≥5.25 lgTCID₅₀/0.1 mL, respectively, and the reductions of PPV in three batches of samples without S/D were 4.57 lgTCID₅₀/0.1 mL、4.18 lgTCID₅₀/0.1 mL、4.68 lgTCID₅₀/0.1 mL, respectively.

With evaluation on the inactivation capability of the indicator viruses, it is proved that the S/D treatment and dry heating has good inactivation and removal effect on the virus in C1-INH.