Objective: To establish a rapid analytical method for chemical components in complex traditional Chinese medicine (TCM) based on an integrated strategy combining ultra high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q TOF MS/MS) with feature-based molecular networking(FBMN), so as to systematically characterize the chemical components in Sishen pills. Methods: MS data for Sishen pills were acquired using UHPLC-Q TOF MS/MS, with separation performed on on a Waters HSS T3 column(2.1 mm×100 mm, 1.8 µm). The mobile phase was consisted of a gradient elution of 0.1% formic acid in water(A) and acetonitrile (B) at a flow rate of 0.3 mL · min^-1. The electrospray ionization (ESI) source was used for both positive and negative ion modes, with a scan range of m/z 80 to 1 500. The data were uploaded to the Global Natural Product Social Molecular Networking (GNPS) to create a FBMN for Sishen pills. Results: A total of 131 compounds were identified in Sishen pills, by analyzing retention time, accurate molecular mass, MS² fragmentation characteristics, comparison with reference standards and self-established database, as well as utilizing FBMN for predicting structural similarity of compounds. These compounds included 27 alkaloids, 63 flavonoids, 11 phenylpropanoids, 14 phenols, 8 terpenoids, and 8 other compounds. Conclusion: In this study, the chemical components of Sishen pills are quickly and comprehensively characterized, which layes a foundation for the effective materials and quality control research of Sishen pills. The study also provides insights for the rapid analysis of chemical constituents in other TCM formulas.

Objective: To develop an inductively coupled plasma mass spectrometry(ICP-MS) method for the determination of Cd, Pb, As, Hg, Co, V, Ni and Se in the crude drug of lanthanum carbonate. Methods: Samples were diluted with 3% nitric acid and directly injected. The matrix effect was eliminated by standard addition method and matrix matching method. Lanthanum oxide was added into the standard solution as the matrix.Compare the linearity, repeatability, accuracy, quantification limit, detection limit and sample determination results of the two methods by methodological validation and sample determination. Results: The linear ranges of ICP-MS standard addition method and matrix matching method were 0-30 ng · mL^-1, the correlation coefficients for all elemental impurities were good(r>0.998). Limits of detection were between 0.003 9-0.22 ng · mL^-1 and 0.009 1-0.72 ng · mL^-1. The RSD of repeatability(n=6) were between 1.5%-4.9% and 5.1%-6.9%. The recoveries were between 87.0%-98.3% and 83.4%-114.4%. The contents of 8 elemental impurities in the crude drug of lanthanum carbonate were basically same. Conclusion: The established two methods can effectively eliminate the matrix effect. They were both simple, rapid, sensitive, accurate and applicable for the elemental impurity control of lanthanum carbonate.

Objective: To establish an HPLC method for the simultaneous determination of chlorogenic acid,puerarin, 3’-methoxy puerarin, puerarin apioside, liquiritin, forsythoside A, 3,5-O-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid, forsythin, honokiol and magnolol in Jinlian Quwen capsules. Methods: The analysis was performed on a Waters Atlantis^TM T3 (150 mm×4.6 mm, 3 μm) column, and the mobile phase was comprised of acetonitrile-0.1% phosphoric acid, with the flow rate of 1.0 mL · min^-1 in a gradient elution manner. The column temperature was set at 35 ℃. The injection volume was 4 μL and the UV detection wavelengths were set at 225 nm (detecting forsythin), 250 nm (detecting puerarin, 3’-methoxy puerarin and puerarin apioside), 276 nm(detecting liquiritin), 290 nm (detecting honokiol and magnolol) and 325 nm (detecting chlorogenic acid,3,5-O-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and forsythoside A). Results: All the 11 constituents showed good linearity within their detection ranges (r≥0.999 5), whose average recoveries (n=6) were 100.1%-108.1%, with the RSDs of 2.8%-4.1%. The content ranges of 11 components in three batches of samples were 7.505-7.606 mg · g^-1, 3.485-3.920 mg · g^-1, 0.969-1.068 mg · g^-1, 0.837-0.955 mg · g^-1, 1.009-1.436 mg · g^-1,3.037-3.602 mg · g^-1, 5.259-5.371 mg · g^-1, 0.931-1.012 mg · g^-1, 1.428-2.053 mg · g^-1, 0.939-1.018 mg · g^-1 and 2.744-2.827 mg · g^-1, respectively. Conclusion: The method is simple, sensitive and reliable. It can be used as a reference for the establishment of quality standard of Jinlian Quwen capsules.

Objective: To meet the needs of gadolinium-based contrast agent (GBCA) analysis and testing, and to ensure drug quality and safety, a new strategy for inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) determination of 12 impurity elements (Cr, Fe, Co, Ni, Cu, V, Zn, As, Cd, Ba, Sb, and Pb) in GBCA was proposed. Methods: GBCA was diluted with 2% HNO₃ and directly analyzed by ICP-MS/MS. Spectral interferences from V, Cr, Fe, Co, Ni, and Cu were eliminated using cool plasma/NH₃/He reaction mode. In addition,spectral interferences from Zn, As, Cd, Ba, and Sb were eliminated using hot plasma/O₂/H₂ reaction mode. To evaluate the accuracy and reliability of the method, the standard reference material (NIST SRM 1643f) was analyzed by comparing the method with high-resolution ICP-MS (HR-ICP-MS), and the spiked recovery experiment was conducted. Results: Under optimized conditions, the limits of detection (LOD) of the analytes were 0.014-0.86 ng · L^-1, the spiked recoveries were 96.7%-105.8%, and the RSDs were 1.6%-4.0%. The HR-ICP-MS determination values were consistent with reference values of NIST SRM 1643f. Statistical analysis showed that there was no significant difference between the measured values of the two methods at a 95% confidence level. Conclusion: The analysis method is accurate, reliable, stable and high precision. The combination of ICP-MS/MS and various reaction modes under different plasma conditions has shown promotion value for multi-element analysis in GBCA and can be extended to other fields.

Objective: To compare the chemical composition changes in Kehuang before and after fermentation using multi-component content determination combined with chemometric analysis. Methods: HPLC was employed using a ZORBAX Eclipse Plus C₁₈ column (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile and 0.1% phosphoric acid aqueous solution, applied with gradient elution. The flow rate was set at 1.0 mL · min^-1,the column temperature was maintained at 30 ℃, the detection wavelength was 203 nm, and injection volume was 10 μL. This method was applied to analyze the chemical compositions of Kehuang before and after fermentation and to establish their corresponding chemical fingerprint profiles. Quantitative determination was conducted for 11 chemical constituents, including chlorogenic acid, cimifugin, notoginsenoside R₁, baicalin, berberine, quercetin,baicalein, wogonin, emodin, chrysophanol, and physcion. Furthermore, chemometric methods such as hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to distinguish and compare the samples before and after fermentation, with the aim of identifying key differential compounds. Results: Chemical fingerprint profiles were established for Kehuang before and after fermentation, with 22 common peaks identified. The fingerprints had good specificity and can be used for quality evaluation of Kehuang. The results of multi-component content determination and chemometrics analysis revealed significant differences in the content of notoginsenoside R₁ (1.023-2.927 mg · g^-1),scutellarein (0.125-0.568 mg · g^-1), and berberine hydrochloride (2.151-3.068 mg · g^-1) among the crude Kehuang powder, fermented Kehuang powder, and the content of Kehuang capsules. Notoginsenoside R₁, scutellarein and berberine were identified as differential markers between pre- and post-fermentation of Kehuang, which could serve as quality indicators for distinguishing and identifying Kehuang before and after fermentation. Conclusion: The chemical composition of Kehuang changes significantly after fermentation, and the differential compounds have been clearly identified, which provides an analytical basis for quality control in the production of Kehuang capsules.

Objective: To establish and validate a double antibody sandwich ELISA method for the in vitro relative potencytestof severe acute respiratory syndrome coronavirus 2, (SARS-CoV-2) recombinant protein vaccine(CHO cells). Methods: Human monoclonal antibodies GH4 and CB6 against the receptor binding domain (RBD)of SARS-CoV-2 spike protein were prepared by genetic recombinant technology. A double antibody sandwich ELISA method was established using GH4 as coating antibody and CB6 labeled with HRP enzyme (CB6-HRP) as detection antibody. The working concentrations of GH4 and CB6-HRP were determined. Methodological validation was carried out,including linearity and range, specificity, precision (repeatability, intermediate precision), accuracy, and robustness. The established method was used to detect the in vitro relative potencies of three batches of process-validated and twenty-two batches of continuous process verification of SARS-CoV-2 recombinant protein vaccines (CHO cells). Results: The optimal working concentrations of GH4 and CB6-HRP were 1 000 ng · mL^-1 and 31.25 ng · mL^-1, respectively. The vaccine reference had a good log-linear relationship with A₄₅₀ in the concentration range of 0.312 5-5 ng · mL^-1,and the slopes of the regression equations were all in the range of 0.80-1.25 and the R² values were all>0.99.The established method could specifically detect the SARS-CoV-2 recombinant protein vaccine (CHO cells), and there was no cross-reactivity with the MERS Vaccine (CHO cells), influenza virus vaccine, rabies virus vaccine and CHO host cell proteins. For precision validation, the geometric coefficient of variation (GCV) of repeatability was in the rang of 1.6% to 2.4%, and the GCV of intermediate precision was in the range of 1.2% to 2.6%. The R² value of the regression equation was 0.994 5, and the slope was 0.999 5 which was in the range of 0.80-1.25. For accuracy validation, the relative bias (RB) was in the range of -4.04% to 7.36%. In the robustness validation, the in vitro relative potency under different test conditions ranged from 0.96 to 1.08. The geometric mean in vitro relative potency of the three batches of process-validated SARS-CoV-2 recombinant protein vaccines (CHO cell) was 1.02, with an GCV of 4.8%; the geometric mean in vitro relative potency of twenty-two batches of continuous process verification SARS-CoV-2 recombinant protein vaccines (CHO cells) was 1.04, with an GCV of 4.4%. Conclusion: The established double antibody sandwich ELISA method has good specificity, precision, accuracy and robustness. It can be used for the detection of the in vitro relative potency and the quality control of SARS-CoV-2 recombinant protein vaccine (CHO cells).

Objective: To characterize the primary structure of the active national standard candidate of infliximab to provide a technical basis for this monoclonal antibody. Methods: The complete relative molecular mass and the relative molecular masses of the light and heavy chain subunits of the active national standard candidate of infliximab were detected based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS). The relative molecular masses of the light and heavy chain subunits of the active national standard candidate of infliximab were analyzed by UPLC-MS/MS. UPLC-MS/MS was used to analyze the sequence coverage,glycosylation sites and disulfide bond localization of infliximab activity candidate national standard products. Results: The complete relative molecular mass of infliximab was 148 515. The relative masses of its light and heavy chain subunits were 23 435 and 50 827 respectively. The sequence coverage of infliximab was 100% for light chain and 100% for heavy chain. The glycosylation sites of infliximab were analyzed by UPLC-MS/MS; and the disulfide bonds were localized. The glycosylation site of the monoclonal antibody candidate was located at N₃₀₀ of the heavy chain; there were 16 pairs of disulfide bonds, 4 pairs within each of the two heavy chains, 2 pairs between chains,2 pairs within each of the 2 light chains, and 1 pair between each of the two groups of light and heavy chains. Conclusion: The primary structural confirmation of the national standard candidate of infliximab monoclonal antibody activity provides a technical guarantee for the establishment of the monoclonal antibody standard substance, the standardization of analytical methods and the control of product quality.

Objective: To establish the HPLC fingerprint of Gardeniae Fructus before and after processing, and to study the relationship between the fingerprint and antioxidant activity. Methods: Agilent 1200 high-performance liquid chromatograph was used, and the column was Agilent ZORBAX SB-C₁₈ (250 mm×4.6 mm, 5 μm). The mobile phase system was 0.1% formic acid water-methanol solution. The ultraviolet detection wavelength was 254 nm. The column temperature was 25 ℃. The extraction solvent of the test solution was 70% methanol solution.The injection volume was 5 μL. The flow rate was 1 mL · min^-1, and the fingerprints of six batches of gardenia before and after processing were established, and the antioxidant activity of gardenia before and after processing was evaluated by DPPH method, and the spectral effect relationship was studied by grey correlation analysis and partial least squares regression analysis. Results: 23 common peaks were identified in the fingerprint, and five chromatographic peaks were identified by comparison of reference substances, including geniposidic acid (peak 3), genipin gentiobiside (peak 9), gardenside (peak 11), crocetin I (peak 21), and crocetin Ⅱ (peak 22). The mean half inhibitory concentration (IC₅₀) of DPPH radical in 5 different processed products (raw gardenia, fried gardenia,wine gardenia, gardenia charcoal and ginger gardenia) were 0.25 mg · mL^-1, 28.61 mg · mL^-1, 6.34 mg · mL^-1,11.79 mg · mL^-1, and 0.68 mg · mL^-1, respectively. Combined with the three statistical methods, peaks 3, 9, 11, 21,and 22 were the common peaks associated with DPPH radical scavenging. Conclusion: The antioxidant activity of 6 batches of Gardeniae Fructus before and after processing is the result of the interaction of multiple components, and peaks 3, 9, 11, 21, and 22 are closely related to antioxidant activity. The results of this study provide a reference for the basic research and clinical application of antioxidant substances in Gardeniae Fructus before and after processing.

Oligosaccharides are a class of bioactive components abundantly found in Chinese herbal medicine. They have clinical effects such as antidepressant and anti-Alzheimer’s disease activities, as well as pharmacological activities such as hypoglycemic, laxative, and immune-enhancing effects. The bioactivity of oligosaccharides is closely related to their structure. However, due to the diversity of glycosidic bond configurations, monosaccharide compositions and linkage patterns, it is difficult to accurately determine their structures by conventional analytical methods. Establishing rapid and reliable analytical methods remains a key challenge in carbohydrate research.Although studies on oligosaccharides are increasing, there is a lack of comprehensive reviews on the structure and analytical methods of oligosaccharides in Chinese herbal medicine. This paper systematically reviewed the research progress over the past decade on extraction, separation, structure identification and analytical methods of oligosaccharides in Chinese herbal medicine by consulting domestic and foreign literatures. This review aims to contribute to the development of evaluation method of traditional Chinese medicine based on oligosaccharide components, and provide reference for their application in the quality control of traditional Chinese medicine.

Objective: To investigate the improvement in bitter taste of ambroxol hydrochloride by the corrective materials and weight gain of the taste-masking layer, and to establish an objective and scientific taste evaluation method. Methods: The taste-masking effects of corrective materials and taste-masking layer weighting were evaluated using electronic tongue technology, and the palatability between ambroxol hydrochloride direct oral granules and 15 marketed preparations of ambroxol hydrochloride were compared. The electronic tongue datas were subjected to principal component analysis and loadings analysis. Results: The corrective materials were able to mask the bitter taste of ambroxol hydrochloride. The bitter value of the sample solution of ambroxol hydrochloride direct oral granules decreased as the masking layer gained weight. In terms of bitterness and sweetness, ambroxol hydrochloride oral direct granules had a clear advantage over 15 marketed formulations. Conclusion: Using the electronic tongue technology, the taste correction effect and the taste difference between different samples can be accurately assessed,and an evaluation method for the bitter taste masking effect of ambroxol hydrochloride direct oral granules has been established, which provides new ideas and methods for the taste masking study of oral preparations for children.