Objective: To establish an ultra-high performance liquid chromatography tandem mass spectrometry（UPLC-MS/MS）method for the simultaneous determination of 22 components of Bimin capsules, including benzoylaconine, benzoylmesaconine, benzoylhypacoitine, hypaconitine, mesaconitine, fuziline, aconitine, aconine,mesaconine, isoliquiritigenin, echinatin, protocatechuic acid, liquiritigenin, isoliquiritoside, glycycoumarin,naringenin, schaftoside, licochalcone B, glycyrrhizic acid ammonium salt, liquiritin, liquiritin apioside and isoliquiritin apioside. Methods: The Acclaim^TM RSLC 120C₁₈ column (2.1 mm×100 mm, 2.2 μm) was used with 0.05% formic acid water (A) and acetonitrile (B) as the mobile phase at a flow rate of 0.4 mL · min^-1. Electrospray ionization (ESI) source and multiple reaction monitoring (MRM) mode in both positive and negative were used for ion detections. Resulets:The 22 components displayed a good linear relationship (R²>0.990) in their respective ranges, and the precision, repeatability, stability, and sample recovery rate all met the requirements. It was found that all 22 components were detected in the nasal sensitivity capsules, with significant differences in the content of some components. Conclusion: This method is fast, simple, sensitive and reliable, and can provide scientific basis for the quality control of the Bimin capsules.

Objective: To explore the growth trend of volatile components in hawthorn fruit at different growth stages. Methods: The volatile components in hawthorn fruits at different growth stages were quantitatively and qualitatively analyzed by gas chromatography-mass spectrometry (GC-MS), and their dynamic changes were investigated and statistically analyzed. The chromatographic column used was HP-5 (30 m×0.32 mm, 0.25 μm),while the column temperature was programmed (initial temperature 60 ℃, increased to 250 ℃ at a rate of 10 ℃ · min^-1,maintained for 33 min), and the inlet temperature was 280 ℃. The ion source was an electron bombardment source(EI), with an ion source temperature of 230 ℃. Results: A total of 57 representative volatile components were detected,of which 16 components disappeared in the later stage of growth, and 12 components were not present in the early stage of growth. The content of 11-decyl-tetracosane was the highest and gradually increased, and the content of hexadecyl acrylate gradually decreased. Conclusion: The investigation of the dynamic changes of volatile components in hawthorn fruit during the growth period provides a reference for further rational development and utilization of hawthorn fruit.

Objective: To establish a method for determining the fingerprint of Yankening tablets and evaluate their quality combined with multivariate statistical analysis and multi-component quantification. Methods: The seperation was performed on a 30 ℃ thermostatic InertSustain C₁₈(250 mm×4.6 mm, 5 μm) column,with the mobile phase comprising of methanol-acetonitrile-0.2% phosphoric acid flowing at 1.0 mL · min^-1 in a gradient elution manner, and the detection wavelength was set at 254 nm to establish HPLC fingerprint of Yankening tablets,contents of 10 components (berberine hydrochloride, baicalin, wogonoside, baicalein, aloe emodin, wogonin, rhein,emodin, chrysophanol, emodin methyl ether) in 17 batches of Yankening tablets were determined, and similarity evaluation, cluster analysis, entropy weight TOPSIS method, and entropy weight grey correlation analysis were used to comprehensively evaluate the quality of Yankening tablets. Results: There were significant differences in the quality of Yankening tablets of the current market. 33 common peaks were obtained by matching in the fingerprints of 17 batches of samples, and the similarity evaluation and cluster analysis results showed differences among different samples. The comprehensive quality ranking and multi-component content determination revealed that the important factors affecting drug quality were the quality and process control level of the three medicinal herbs,Phellodendri Chinensis Cortex, Coptidis Rhizoma and Scutellariae Radix. Conclusion: The fingerprint established by this method, combined with a multivariate statistical model, can provide reference for the quality evaluation research and products quality improvement of Yankening tablets.

Objective: To establish a multi-component determination method and HPLC fingerprint for Vernonia amygdalina Del., combined with chemical pattern recognition to comprehensively evaluate the quality of Vernonia amygdalina Del. from different producing, laying a foundation for the future development of quality control standards for Vernonia amygdalina Del. and other related research on Vernonia amygdalina Del.’s components. Methods: The HPLC using Welch Ultimate^® AQ-C₁₈ column (250 mm×4.6 mm,5 µm) and 0.2% phosphoric acid aqueous solution - acetonitrile as moblie phase by gradient elution at the flow rate of 1 mL · min^-1, wavelength was set at 258 nm and the column temperature was 35 ℃. Simultaneously establish 24 batches of fingerprint spectra of Vernonia amygdalina Del. from different origins and a multi-component HPLC determination method for 9 components of Vernonia amygdalina Del.. Combine chemical pattern recognition to evaluate the quality of 24 batches of Vernonia amygdalina Del. from different origins. Results: The similarities of the fingerprint spectra of 24 batches of Vernonia amygdalina Del. were above 0.9. The established control fingerprint spectra could stably and effectively distinguish Vernonia amygdalina Del. qualitatively. There were 10 common peaks in the fingerprint spectra, and 9 peaks were identified. The mass fractions of 24 batches of Vernonia amygdalina Del. components that were uracil, chlorogenic acid, luteolin, luteolin-7-O-β-D-glucuronide, isochlorogenic acid B, 3,5-O-dicaffeoyl quinic acid, 1,5-dicaffeoyl quinic acid, apigenin-7-O-β-D-glucopyranoside, 4,5-O-dicaffeoyl quinic acid were 0.007 314–0.084 30 mg · g^-1，0.619 6–9.763 mg · g^-1，0.303 8–4.031 mg · g^-1，0.984 6–8.146 mg · g^-1，0.043 29–0.438 7 mg · g^-1，0.537 5–11.57 mg · g^-1，0.437 6–13.78 mg · g^-1，0.032 19–0.720 1 mg · g^-1，0.190 0–1.931 mg · g^-1. Through cluster analysis, 24 batches of Vernonia amygdalina Del. were divided into 2 categories. Vernonia amygdalina Del.from Guangdong and Hainan were classified as one category, while Vernonia amygdalina Del. from Guangxi were classified as one category alone. The classification results of principal component analysis were consistent with cluster analysis. Further research on the differences in leaf quality among different regions of Vernonia amygdalina Del. was conducted through principal component scores, and it was found that the P5, P24 and P22 regions ranked in the top three with better quality than other regions. Through orthogonal partial least squares discriminant analysis, it was found that the reasons for the differences in leaf quality among different regions of Vernonia amygdalina Del. were ranked in descending order: 1,5-dicaffeoyl quinic acid, chlorogenic acid, and luteolin-7-O-β-D-glucuronide, 3,5-O-dicaffeoyl quinic acid. Conclusion: The fingerprint and multi-component determination methods of Vernonia amygdalina Del. are stable and effective through methodological detection, which can fill the gap in quality control standards for Vernonia amygdalina Del.. Combined with chemical pattern recognition methods, the high-quality production areas of Vernonia amygdalina Del. and the components that should be emphasized in controlling the quality of Vernonia amygdalina Del. from different production areas are pointed out.A comprehensive evaluation of the quality of Vernonia amygdalina Del. has been conducted, providing a solid foundation for future quality control and evaluation of Vernonia amygdalina Del..

Objective: To establish the UHPLC fingerprint and quantitative analysis of CBDA, CBD, THC, and THCA for hemp, and provides a basis for the quantity control of hemp. Methods: UHPLC was launched on a Phenomenex C₁₈ (150 mm×4.6 mm, 2.6 μm) with mobile phase of acetonitrile-0.2% phosphoric acid aqueous solution at a flow rate of 1.0 mL · min^-1, column temperature of 35 ℃, detection wavelength of 220 nm, and an injection volume of 2 μL by gradient elution. 15 batches of hemp samples from Daxing ’anling region and Qiqihar were analyzed to establish the fingerprint. Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were adopted to assess of UHPLC fingerprint of hemp. Results: the UHPLC fingerprint of hemp was established, and 26 common peaks were selected the 26 common peaks were compared with the reference substance. The structures of four chromatographic peaks were identified and simultaneously analyzed for content determination. four chromatographic peaks were identified as CBDA, CBD, THC and THCA.The similarity of fingerprint of them was greater than 0.950. The samples from Daxing’anling were clustered into class Ⅰ, with the best quality. The results show that hemp samples from different regions were quite different. The variable importance projection (VIP) showed that the four compounds to be tested were the main components leading to the difference of hemp. Conclusion: The established method is stable and reliable and can be used for qualitative and quantitative analysis of hemp. The findings of this study are expected to provide a scientific basis for quality control of hemp.

Objective: To extract and isolate polysaccharides from Green Fructus Forsythiae and Folium Forsythiae for structural characterization and comparison of their anti-inflammatory activities. Methods: Crude polysaccharides were obtained through extraction and purification processes. Gel permeation chromatography and laser light scattering were used to determine their molecular weight. The MTT assay was employed to evaluate the cytotoxicity of the polysaccharides, while ELISA was used to measure levels of nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), and interleukin-10 (IL-10). Results: The polydispersity index (PDI) indicated a relatively uniform molecular weight distribution. Monosaccharide composition analysis showed that both polysaccharides contained various monosaccharides, with Green Fructus Forsythiae polysaccharide having a higher content of galacturonic acid, while Folium Forsythiae polysaccharide had a higher glucose content. Methylation analysis revealed that polysaccharides in Green Fructus Forsythiae were primarily linked by 4-Gal (p)-UA, while those of Folium Forsythiae polysaccharide were mainly linked by 4-Glc (p). Both polysaccharides significantly inhibited the production of NO and TNF-α induced by LPS while promoting the generation of IL-10, with polysaccharide in Folium Forsythiae showing superior effects compared to Green Fructus Forsythiae polysaccharide. Conclusion: Polysaccharides from Green Fructus Forsythiae and Folium Forsythiae exhibit significant anti-inflammatory activities. This research lays the foundation for the development of Folium Forsythiae resources.

Objective: To establish a recombinant C-factor method to detect the content of bacterial endotoxin in recombinant type Ⅲ humanized collagen solution for injection. Methods: Three batches of injectable recombinant type Ⅲ humanised collagen solutions were determinedand method validation by using the BIOMERIEUX recombinant C-factor kit and the lonza recombinant C-factor kit，respectively. Results: The results showed that about the two kits the concentration points of the standard curve were ≥ 3, and the linear correlation coefficient was r > 0.980,the ΔRFU value of the negative control was smaller than that of the lowest point of the standard curve; the bacterial endotoxin contents were all < 6 EU · mL^-1. The reproducibility was good, the recoveries of bacterial endotoxin were in the range of 50%-200%, which was in accordance with the standard requirements of the ChP. Conclusion: The bacterial endotoxin content in the solution of recombinant type Ⅲ humanized collagen solution for injection is determined by the two kits. The results were < 0.05 EU · mL^-1 and < 0.005 EU · mL^-1, which are in accordance with the standard requirements for bacterial endotoxin in the product, respectively. The method can be used for the determination of bacterial endotoxin content in the solution of recombinant type Ⅲ humanized collagen solution for injection. This study provides a reference for the research related to the determination of endotoxin in recombinant protein products using recombinant C-factor method.

Objective: To establish an inductively coupled plasma mass spectrometry (ICP-MS) method to determine the contents of 10 elemental impurities in the pharmaceutical excipients sodium succinate according to the ICH Q3D elemental impurity guideline, and to provide a basis for the comprehensive evaluation of potential risks. Methods: According to the types of elements that needed to risk assessment and the permitted daily exposure (PDE) of the injection route, the single-component limit calculation method was used to calculate the limit and control threshold of each element in sodium succinate with 10 g · d^-1 as the maximum daily dose.Samples were preprocessed by direct dissolution and the iCAP RQ ICP-MS was used for the simultaneous determination of the residual amounts of 10 impurities in sodium succinate, using Ge, In and Bi as the internal standards. Results: All elemental impurities showed good linear relationship (r>0.99). The limits of detection of Cd, Pb, As, Hg, Co, V, Ni, Li, Sb and Cu were 0.27，19.22，8.86，16.63，0.12，0.28，1.48，6.91，0.35，9.26 ng · g^-1, respectively. The recoveries (n=3) of each concentration were between 82.4% and 130.9%, and the RSD (n=6) of the repeatability test was not more than 7.1%, with all findings meeting the requirements for methodological validation. The content of elemental impurities in sodium succinate was less than 30% of the PDE,indicating that the elemental impurities in sodium succinate had no potential safety risk with the medicinal product. Conclusion: The method is simple in sample pretreatment, with high sensitivity and good accuracy. It is suitable for monitoring and risk assessment of elemental impurities in sodium succinate, which is conducive to the quality control. It also provides a reference for the determination of elemental impurities in other products.

Objective: To analyze six components in Bletillae Rhizoma using high-performance liquid chromatography. Through the linear calibration using two reference substances method, the retention time of the components was predicted, and the feasibility and accuracy of this technique in the qualitative identification of chromatographic peaks were explored. This method was also compared with the relative retention time (RRT) method. Methods: High-performance liquid chromatography was employed using an Ultimate XB-C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of water (A) and acetonitrile (B), with a gradient elution as follows: 0-11 min,12% B; 11-58 min, 12% B to 68% B; 58-65 min, 68% B to 12% B. The flow rate was 0.9 mL · min^-1, the injection volume was 5 μL, the column temperature was maintained at 30 °C, and the detection wavelength was set at 280 nm. By measuring the actual retention time of the six components on 16 different C₁₈ chromatographic columns, the standard retention time was calculated. Gastrodin (Peak 1) and gymnadenin Ⅲ (Peak 4) were used as the double-standard compounds.The linear calibration using two reference substances method was used to predict the retention time of other components,and the prediction was verified on another three unknown C₁₈ chromatographic columns. With the relative retention time(RRT) method, using bletilloside (Peak 5) as the reference compound, the retention times of the other five components were predicted. Results: A qualitative analysis method for the six components in Bletillae Rhizoma was established,which could accurately predict the retention times of each component in Bletillae Rhizoma. The standard retention times of the six components as as follows: gastrodin 4.279 min, gymnadenin Ⅲ 32.145 min, pleione bulbocodioides factor Ⅰ27.220 min, 2-O-glucosylbletilloside 28.603 min, bletilloside 33.509 min, and batatasin Ⅲ 45.504 min. Conclusion: The linear calibration using two reference substances method can improve the prediction ability of the retention time of each component in Bletillae Rhizoma, thereby enhancing the accuracy and reliability of the analysis results.

Objective: To establish, validate and apply an enzyme-linked immunosorbent assay (ELISA)method for determination of residual human factor Ⅻ(FⅫ) and prekallikrein (PK) in high concentration human intravenous immunoglobulin (10%IVIG). Methods: Microplates were pre-coated with specific antibodies for FⅫ and PK, respectively, and then blocked. The FⅫ and PK in 10%IVIG were allowed to bind with the pre-coated antibodies on the plates. After appropriate washing steps, biotinylated detection antibodies specific for FⅫ and PK were added. Following a second washing, HRP/Streptavidin-Peroxidase conjugates were added and unbound conjugates were washed away with another washing step. After the addition of a chromogenic substrate and a stop solution, the absorbance was measured at 450 nm using a microplate reader. The detection method was validated according to the 2020 edition of Chinese Pharmacopoeia. Then the validated method was used to determine the residual levels of FⅫ and PK in process intermediates and final products 10%IVIG. Results: The dilution linearity results showed a matrix effect and a hook effect in FⅫ and PK detection methods, respectively,which could be solved by diluting 5-20 times and 80-120 times, respectively. The average recoveries in the spiked experiments were 110.3% and 99.1% for FⅫ and PK respectively, with RSD of 3.6% and 2.7%. indicating good accuracy of the method. The RSDs of the repeatability validation were 8.3%、7.7% for both methods, and the RSD of intermediate precision validation were 7.0% and 9.2% for FⅫ and PK, respectively, indicating good precision of the two methods. The limits of quantitation of the two methods were 1.0 ng · mL^-1 and 1.25 ng · mL^-1 respectively.Both methods exhibited good linearity (r = 0.999 9). The robustness validation, which included incubation time,the effective period of the opened reagent kit, and different batches of reagent kits, showed good robustness of the method. The residues of high concentration human immunoglobulin FⅫ and PK maintained at low levels by using the method, indicating that octanoic acid precipitation combined with two-step anionic exchange chromatography process could effectively remove FⅫ and PK in 10%IVIG. Conclusion: The detection method shows good applicability and can be used for the determination of FⅫ and PK residues in 10%IVIG.