Objective: To develop a biological activity reference material for recombinant COVID-19 vaccine (CHO cell) for the evaluation of in-vitro relative potency and in-vivo potency in mice for recombinant COVID-19 vaccine (CHO cell). Methods: One qualified bulk of recombinant COVID-19 vaccine (CHO cell) was selected. After calibration of protein content, an aluminum adjuvant was added in the bulk to prepare one batch of vaccine as the candidate biological activity reference material. After passing the finished product test, the candidate biological activity reference material was subjected to homogeneity tests. Five independent experiments using one batch of phase Ⅲ clinical vaccine as the biological activity reference material were conducted to calibrate the in-vitro relative potency and in-vivo potency in mice for candidate biological activity reference material. The stability of the candidate biological activity reference material was analyzed by storage at (37±2) ℃ and (5±3) ℃. The phase Ⅲ clinical batch vaccine and the candidate biological activity reference material were used to test the applicability of the candidate biological activity reference material by evaluation of the in-vitro relative potency and in-vivo potency of six batches of vaccines. Results: All test items of the biological activity reference material for the recombinant COVID-19 vaccine (CHO cell) complied with the requirements. The filling volume was not less than the labeled amount, with the filling precision within ±1%. The RSD values of D10, D50, and D90 reflecting particle size distribution were 0.45%, 0.39%, and 0.54%, respectively, and that of antigen content was 2.7%. The phase Ⅲ clinical vaccine, when being calibrated as a candidate reference material, demonstrated the in vitro relative potency and in-vivo potency in mice no less than 0.5, with the geometric mean values of 1.0 and 1.0 and the geometric coefficients of variation of 8.9% and 31%, respectively. After storage at (37±2) ℃ for 28 days and (5±3) ℃ for 36 months, the in-vitro relative potency and the in-vivo potency in mice showed no significant upward or downward trends, indicating good stability. The results of the in-vitro relative potency and in-vivo potency in mice tests for the six batches of vaccines using phase Ⅲ clinical vaccine and the candidate biological activity reference material ranged from 0.9 to 1.0 and 0.7 to 1.4, respectively, indicating former and candidate biological activity reference materials had good consistency. Conclusion: The biological activity reference material for recombinant COVID-19 vaccine (CHO cell) can be used for evaluation of the in-vitro relative potency and the in-vivo potency in mice for this vaccine.

Focusing on the quality control items specified in the quality standards for antibody products, this paper systematically reviews the latest advances in analytical technologies, encompassing the detection of key quality attributes such as size heterogeneity, charge heterogeneity, glycan heterogeneity, biological activity (including binding/functional activity assays and reporter gene-based methods), excipient content, protein content, peptide mapping analysis, and the quantification of host-related residual impurities. The detection principles, key considerations for method development, and applicable scenarios of the aforementioned technologies are detailed. In addition, the technical advantages, core development points, and application progress of multi-attribute monitoring (MAM) technology for the characterization of post-translational modifications are expounded. The quality control methodology system and technical advances of antibody products summarized in this study can provide a valuable reference for the establishment of quality standards, optimization of quality control methods, and stability studies of antibody drugs, and further facilitate the improvement of quality control and regulatory standards in the research, development, and manufacturing of antibody products.

Objective: To determine the metabolic profiles of etomidate and its structural analogues metomidate, propoxate, and isopropoxate in human liver microsomes (HLMs). Methods: In a human liver microsome incubation model, four target compounds were separately added and incubated at 37 ℃ for 1 hour. The reaction was terminated by adding ice-cold acetonitrile. After centrifugation, the supernatant was collected, evaporated to dryness under nitrogen, and reconstituted. The processed samples were analyzed using ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry in ESI⁺ mode. The mobile phases consisted of 0.1% formic acid in water and 0.1% formic acid in acetonitrile, with a Waters HSS T3 column used for separation. The IDA scan mode was employed to detect the parent compounds and their metabolites, aiming to investigate the metabolic pathways. Results: The biotransformation of the four compounds primarily involved pathways such as dealkylation, dehydrogenation, oxidation, and glucuronidation. In three real hair samples with positive results, the unchanged forms of etomidate, metomidate, and isopropoxate were detected, along with the same dealkylation metabolites and carboxylation products. Additionally, the characteristic metabolites of two compounds were detected (metabolites M3 yielded from etomidate and isopropoxate after the loss of the phenylethyl group). Conclusion: It is recommended that the unchanged form of etomidate and its metabolite M3, the unchanged form of metomidate, the unchanged form of propoxate and its metabolite M3, as well as the unchanged form of isopropoxate and its metabolite M3 can be used as biomarkers for the intake of the four target compounds.

Objective: To establish an optimized bioassay for bFGF, namely the heparin sodium dilution method, aimed at improving the physiological relevance and stability of testing for heparin sodium-containing preparations. Methods: Based on the method described in General Chapter 3527 of the Chinese Pharmacopoeia (2025 Edition, Part IV), the assay was modified by introducing a maintenance medium supplemented with heparin sodium during the sample dilution step and shortening the cell starvation duration to 5 h. Both the optimized method and the standard pharmacopoeia method were employed to assay bFGF preparations with and without heparin sodium to compare their performance. Results: For heparin-containing preparations, the heparin sodium dilution method showed a significantly higher curve-fitting quality (0.998±0.001) and a wider dose-response interval (0.499±0.070) compared to the pharmacopoeia method (0.995±0.002, p<0.01, and 0.295±0.039, p<0.000 1, respectively). The EC₅₀ value (0.419±0.088) was significantly lower than that of the pharmacopoeia method (0.627±0.154) (p<0.000 1). No significant difference was observed in labeled potency between the two methods, and the optimized method demonstrated a satisfactory linear range. For heparin-free preparations, both methods met the correlation coefficient requirement (r>0.9) and showed good parallelism; however, the labeled potency measured by the heparin sodium dilution method (146.78%±10.0%) was significantly higher than that of the pharmacopoeial method (90.65%±2.8%) (p<0.001). Conclusion: The heparin sodium dilution method improves the fitting quality, sensitivity, and method stability of the four-parameter regression assay for heparin sodium-containing preparations. It serves as a valuable supplement to the pharmacopoeia method. These findings provide a more scientific and reliable approach for the quality control of bFGF preparations.

Objective: To establish a cell line and detection method based on the reporter gene assay (RGA) for evaluating the biological activity of denosumab. Methods: The RANK-GloResponse NF-κB-RE-luc2P HEK293 cell line was constructed by transfecting HEK293 cells with plasmids carrying the RANK gene and the NF-κB response element (NF-κB-RE) coupled with a luciferase reporter gene (Luc2P) via transgenic technology. After optimizing experimental conditions including cell density, chromogenic assay conditions, quantity of RANK ligand (RANKL), and denosumab dilution gradient, the detection method was established. The accuracy, precision, linear range, specificity, and robustness of the method were validated in accordance with 9101 and 9401 of General Rules of the Pharmacopoeia of the People’s Republic of China, Four Volumes. Results: The RGA for detecting the biological activity of denosumab was successfully validated. Within the potency level range of 64%-156%, the relative bias (RB) of the method was within±12.0%. The regression equation had a slope of 1.047 6 and a correlation coefficient of 0.988 1. The maximum coefficient of variation (RSD) for precision and accuracy was 15.0%, and the maximum geometric coefficient of variation (GCV) for robustness across different cell passages, cell densities, and culture times was 7.9%. Cell passage stability tests confirmed that the probability (P) value exceeded 0.05 for up to 30 generations. Conclusion: This study successfully constructed a reporter gene-based cell line suitable for detecting the biological activity of denosumab. Methodological validation demonstrated its reliability, providing a robust technical support for the quality control of related biological products.

Objective: To establish reversed-phase high-performance liquid chromatography (RP-HPLC) and dual-wavelength size exclusion high-performance liquid chromatography (SE-HPLC) methods for effectively analyzing the conjugation status of simulated deconjugated antibody-drug conjugate (ADC) formulations, and to combine these methods with ultraviolet (UV) spectrophotometry for protein concentration determination, aiming to meet the requirements of formulation analysis. Methods: Group A formulations simulating partially deconjugated ADC were prepared by physically mixing intact ADC, small-molecule drug, and antibody. Corresponding control Group B formulations, without simulated deconjugation but with matched total antibody and total small-molecule drug concentrations, were also prepared. A UV spectrophotometric method was established to measure absorbance of Group A and B formulations at 252 nm and 280 nm. The drug-to-antibody ratio (DAR) was calculated according to the Beer-Lambert law to reflect the total antibody concentration and conjugation status. An RP-HPLC method was developed by gradient elution through a Zorbax Ellipse XDB-C₁₈ column (50 mm×2.1 mm, 3.5 μm) at 35 ℃ with 0.1% trifluoroacetic acid in water as mobile phase A and 0.08% trifluoroacetic acid in acetonitrile as mobile phase B at the flow rate of 0.5 mL · min^-1, autosampler temperature of 5 ℃, and detection wavelength of 252 nm. This method was used to detect free small molecules generated from deconjugation, indirectly analyzing the conjugation status of ADC. A dual-wavelength SE-HPLC method was established by elution through a TSK G3000SWXL gel column (7.8 mm×30 cm, 5 μm) with the mobile phase consisting of 15% isopropanol and 85% phosphate buffer (0.2 mol · L^-1 potassium phosphate, 0.25 mol · L^-1 potassium chloride, pH 7.0) at room temperature, the flow rate of 0.5 mL · min^-1, autosampler temperature of 5 ℃, and detector wavelengths of 252 nm and 280 nm. This method was used to directly calculate the DAR of ADC formulations by measuring the corresponding peak areas. Results: The total antibody concentrations in Group A and B formulations determined by UV spectrophotometry (n=3) were 5.14 mg · mL^-1 and 5.04 mg · mL^-1, respectively, with corresponding DAR values (n=3) of 3.45±0.03 and 3.47±0.02. The RP-HPLC method effectively detected the simulated free small molecules in Group A formulations, which were absent in Group B. The DAR values determined by dual-wavelength SE-HPLC for Group A and B formulations were 1.83 and 3.65, respectively. Conclusion: UV spectrophotometry can accurately determine the antibody concentration in this simulated deconjugated ADC formulation but cannot effectively determine its conjugation status. RP-HPLC can detect free small molecules in this simulated deconjugated formulation, indirectly proving the occurrence of simulated deconjugation. Dual-wavelength SE-HPLC can effectively and accurately determine the conjugation status of this simulated deconjugated formulation. Its combination with UV spectrophotometry for protein concentration determination shows promise in meeting the requirements of formulation analysis.

Objective: To explore the rapid inspection application of three-dimensional fluorescence technology in national drug sampling inspections, organize and construct an innovative technical system integrating high-dimensional spectroscopy and chemometric intelligent modeling, and apply it in quality evaluation of traditional Chinese medicine. Methods: Three-dimensional fluorescence fingerprint spectra of 131 batches of samples, including six traditional Chinese medicines (Aquilariae Lignum Resinatum, Olibanum, Angelicae Dahuricae Radix, Persicae Semen, Stephaniae Tetrandrae Radix and Faeces Trogopterori) and their common adulteration from annual national drug sampling were collected using a three-dimensional fluorescence spectrometer. The Rayleigh scattering and Roman scattering were removes using a piecewise Hermite spline interpolation algorithm. The spectra were factorized by the self-weighting alternating trilinear decomposition (SWATLD) algorithm. Establish three-dimensional fluorescence spectral qualitative models for Aquilariae Lignum Resinatum versus its counterfeit Syringa Pinnatifolia, Olibanum versus its counterfeit Succlnum, Angelicae Dahuricae Radix versus sulfur-fumigated Angelicae Dahuricae Radix, Persicae Semen versus Armeniacae Semen Amarum, Stephaniae Tetrandrae Radix and its counterfeits, as well as Faeces Trogopterori versus artificial Faeces Trogopterori, etc., using various chemometric algorithms such as principal components analysis (PCA), hiearchical cluster analysis (HCA), partial least squares discriminant analysis (PLS-DA), and these models were applied to the rapid inspection and identification of the authenticity, quality, and inferiority of traditional Chinese medicines. Finally, cross-validation was used to evaluate the specificity and sensitivity of the models. Results: The modeling technology based on three-dimensional fluorescence spectra could achieve direct, rapid, green, and near-real-time intelligent qualitative analysis of the quality of traditional Chinese medicines. The correct recognition rate of each model reached 100%. Conclusion: Different traditional Chinese medicines have characteristic three-dimensional fluorescence fingerprint information. The chemometric models established by three-dimensional fluorescence spectra and the SWATLD factor decomposition algorithm can effectively characterize and evaluate the quality differences between traditional Chinese medicines and their common adulteration.

This article traces and compares the origins, distinctions, and interrelationship of the two terms—specificity and selectivity—as used in pharmaceutical analytical methods. It discusses how these terms have been harmonized and incorporated into the guidelines of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). The article also introduces the approaches for evaluating and applying these terms in analytical procedures. Finally, case examples are provided to further illustrate their practical implications and to enhance readers' understanding of the concepts.

Objective: To investigate the microbial contamination of 74 batches of prepared slices of baked licorice and to provide the reference for microbial limit test and criteria of baked licorice. Methods: Referring to the methods in chapters 1105, 1106 and 1108 of the Pharmacopoeia of the People's Republic of China (the Chinese Pharmacopoeia) 2020 edition, 74 batches of samples were examined for total aerobic bacteria (TAMC), total molds and yeasts (TYMC) and total heat-resistant bacteria (HRMC). And the three types of control bacteria, and the typical colonies were identified by using MALDI-TOF-MS, to preliminarily reaveal the level of bioburden of prepared slices of baked licorices and analyze the microbial contamination. Results: The lg values of TAMC in 74 batches of prepared slices of baked licorice ranged from 1.7 to 6.7, with a mean value of 3.6, the lg values of TYMC ranged from 0 to 5.9, with a mean value of 2.3, and the lg values of HRMC ranged from 0 to 6.0, with a mean value of 2.7. Contamination with bile salt resistant gram-negative bacteria was uneven, 43 batches were detected with bile salt resistant gram-negative bacteria. Escherichia coli and Salmonella were not detected in 74 batches, and a total of 18 genera and 44 species of other bacteria were detected. Conclusion: The microbial contamination of prepared slices of baked licorice varies widely and is uncertain, and there is a risk of microbial contamination in production, circulation and use, which is affected by planting and harvesting, preparation process, storage environment and transportation conditions, etc. The pathogenic bacteria detected suggests that the contaminated microorganisms in prepared slices of baked licorice have a certain degree of pathogenicity, and effective measures should be taken to prevent microbial contamination and to establish a reasonable microbial quality standard to improve the quality of prepared slices of baked licorice.

Objective: A method was established for the simultaneous determination of five nitazenes in the blood of mice using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) based on QuEChERS pretreatment technology. Methods: The samples were extracted with a mixture of methanol-acetonitrile (1 ∶ 3, volume ratio) and dehydrated with MgSO₄. The samples were separated on an XSelect HSS T₃ column (2.1 mm×100 mm) by positive ion scanning in multiple reaction monitoring (MRM) mode, with gradient elution using a mobile phase composed of methanol-0.1% formic acid aqueous solution. Results: Five nitazenes exhibited good linearities within the concentration range of 0.1-100 ng · mL^-1, with correlation coefficients all exceeding 0.999 0. The limits of detection (LOD) of five nitazenes were within the range of 0.4-2.0 pg · mL^-1, and the limits of quantitation (LOQ) were within the range of 1.3-6.3 pg · mL^-1. The spiked recovery rates (n=6) ranged from 89.4% to 110%, with relative standard deviations (RSD) of 3.9%-6.3%. Conclusion: This method offers advantages including high sensitivity, accuracy, and strong operability. It can be effectively applied to detect five nitazenes in blood samples, providing a scientific basis for drug-related cases.

Objective: To simultaneously determine the contents of baicalin, berberine hydrochloride, baicalein, wogonin, aloe-emodin, emodin, rhein, chrysophanol and physcion in Yiqing tablets using the quantitative analysis of multi-components by single-marker (QAMS) method. Methods: The HPLC method was employed using an Ultimate^® AQ-C₁₈ column (4.6 mm×250 mm, 5 μm) with a mobile phase of 0.1% phosphoric acid aqueous solution (A)-acetonitrile (B) in gradient elution mode. The analysis was performed at a flow rate of 0.8 mL · min^-1, detection wavelength of 265 nm, column temperature of 30 ℃, and injection volume of 10 μL. Baicalin served as the reference substance for calculating the relative correction factors (RCFs) and determining the contents of the other eight components. Results: Baicalin, berberine hydrochloride, baicalein, wogonin, aloe emodin, emodin, rhein, emodin and emodin methyl ether showed good linear relationships within their respective ranges (r≥0.999 0), with average recovery rates of 95%-110% and RSD<5%. The relative correction factors for berberine hydrochloride, baicalein, baicalein, aloe emodin, emodin, rhein, chrysophanol and emodin methyl ether were 1.71, 1.88, 0.63, 0.78, 0.74, 1.15, 0.80, and 3.65, respectively. The results obtained by QAMS method and external standard method were close to each other. The contents determined by the one QAMS method were 11.93, 6.760, 1.049, 0.135 1, 0.131 6, 0.606 0, 0.153 3 and 0.580 0 mg · g^-1, respectively. Conclusion: This method has good specificity, high accuracy, stability and reliability, and can be used for quality control of Yiqing tablets.

Objective: To establish a probabilistic risk assessment method for non-carcinogenic and carcinogenic risks of heavy metals and harmful elements in Houttuyniae Herba, thus providing guidance for the safe use of traditional Chinese medicine. Methods: The content of lead (Pb), cadmium (Cd), arsenic (As), mercury (Hg), and copper (Cu) in 40 batches of Houttuyniae Herba were measured by inductively coupled plasma mass spectrometry (ICP-MS). On the basis of Monte Carlo simulation, the hazard index (HI) and carcinogenic risk (CR) were employed to assess the non-carcinogenic and carcinogenic probabilistic risks caused by exposure to heavy metals and harmful elements in Houttuyniae Herba. Results: The mean content of Pb, Cd, As, Hg, and Cu in Houttuyniae Herba was 2.62, 0.48, 0.58, 0.04, and 7.95 mg · kg^-1, respectively. The maximum HI values for males and females were 1.44 and 1.39, respectively, indicating that the total non-carcinogenic health risks for high-exposure populations could not be ignored. The carcinogenic risk assessment results indicated that the carcinogenic health risks associated with Pb, Cd, and As in Houttuyniae Herba should be of concern for both general and high-exposure populations. Conclusion: This study explores a probabilistic health risk assessment method for heavy metals and harmful elements in Houttuyniae Herba based on Monte Carlo simulation, considering the characteristics of traditional Chinese medicine consumption. It provides technical support for the safety evaluation of traditional Chinese medicine and the formulation and revision of related limit standards.

Objective: To establish an HPLC method for determination of related substances in abemaciclib. Methods: :The gradient elution was carried out through an InfinityLab Poroshell 120 EC-C₁₈ column (150 mm×4.6 mm,4 μm) with tetrabutylammonium hydroxide aqueous solution (adjusted to pH 10.0 with formic acid) as the mobile phase A and acetonitrile as the mobile phase B at a flow rate of 1.2 mL · min^-1. The detection wavelength was set at 296 nm, and the column temperature was 30 ℃. The injection volume was 10 μL. Results: Abemaciclib can be completely separated from all impurities (separation degree≥2.45). The test solution was stable for at least 7 d. The limits of quantitation of impurities Ⅰ-Ⅶ were all 0.04%. The 7 impurities showed good linearity (r≥0.997 6)in the tested ranges. The average recovery rates of the impurities were in the range of 94.7%-101.7%, with the RSD≤2.2%. The method showed high precision and stability. Conclusion: The method, with high sensitivity, specificity, and accuracy, can be used for the determination of the related substances in abemaciclib.

Amyloid β-protein is one of the core pathological markers of Alzheimer’s disease. The breakthrough of its blood detection technology is of great significance for the early diagnosis of this disease. This paper systematically reviews the current mainstream detection technologies for amyloid β-protein in blood, focusing on the technical principles, detection performance, and clinical application potential of immunoassay-based methods (ELISA, Simoa, electrochemiluminescence, etc.) and mass spectrometry-based methods. Immunoassay-based methods are dominant because of the simple operation and high throughput, while mass spectrometry-based methods have better performance in specificity, accuracy, and multi-subtype detection. Future development needs to focus on standardized process construction, multi-biomarker joint detection, and miniaturization of detection technologies to promote the clinical application of detection technologies for amyloid β-protein in blood.

Objective: To establish an HPLC method for the simultaneous determination of 13 components (loganic acid, chlorogenic acid, caffeic acid, loganin, sylvestrosideⅠ, isochlorogenic acid B, isochlorogenic acid A, cantleyoside, isochlorogenic acid C, dipsacoside B, dipsacoside A, asperosaponinⅩ, and asperosaponinⅥ) in Dipsaci Radix, and to provide a scientific basis for the quality control of Dipsaci Radix. Methods: An Agilent Zorbax SB-C₁₈ column (250 mm×4.6 mm, 5.0 μm) was employed for gradient elution, with 0.05% phosphoric acid solution(A)-acetonitrile (B) as the mobile phase. The flow rate was 0.8 mL · min^-1, the column temperature was 30 ℃, the detection wavelength was 212 nm, and the injection volume was 10 μL. Results: The 13 components showed good linear relationships within the corresponding ranges (r≥0.999 0), with average recoveries from 95.7% to 103.2% and an RSD<2.0%. In the 8 batches of samples, the content ranges of the aforementioned components were 15.92-20.78 mg · g^-1, 2.23-10.56 mg · g^-1, 0.2-0.47 mg · g^-1, 1.64-4.81 mg · g^-1, 0.32-0.70 mg · g^-1, 0.26-1.19 mg · g^-1, 3.32-8.77 mg · g^-1, 16.98-43.50 mg · g^-1, 2.48-6.35 mg · g^-1, 0.29-0.67 mg · g^-1, 1.83-4.39 mg · g^-1, 21.38-39.84 mg · g^-1, and 50.30-102.02 mg · g^-1, respectively. Conclusion: The simple and reliable method can be applied for the content determination and quality control of 13 components in Dipsaci Radix.

This paper systematically sorts out the difficulties in the quality control of recombinant enzyme drugs and conducts an in-depth analysis with five categories of representative enzymes as examples. Thrombolytic enzymes (e.g., tenecteplase) have significant disparities in specific activity due to differences in glycosylation and outdated measurement methods, requiring the establishment of standardized methods and clarification of the determinants of activity. The enzymes (e.g., glucocerebrosidase and α-galactosidase A) for enzyme replacement therapy (ERT) of rare diseases exert their pharmacological effects only through receptor-mediated intracellular transport, requiring the establishment of a multi-dimensional evaluation system from biochemical activity to cellular uptake. Recombinant IgG-degrading enzymes with atypical Michaelis-Menten kinetics pose a fundamental challenge to the conventional evaluation system due to their two-step cleavage mechanism. Process enzymes (e.g.,recombinant nuclease) have long faced the dilemma of no pharmacopoeial standards and no national reference materials, requiring gradual advancement from method establishment, reference material development, collaborative calibration to the formation of national standards. Adjuvant enzymes (e.g., recombinant hyaluronidase) are faced with the lack of national reference materials for recombinant products, and some enterprises directly use extracted heterologous standard substances for activity assignment, resulting in the disordered labeling of activity units. In response to the challenges in activity evaluation of the aforementioned recombinant enzyme drugs and key enzyme-based raw and auxiliary materials and through a review of issues identified during the registration testing of representative drugs, findings from systematic research projects, and literature on drug evaluation strategies, this article summarizes key technical points for the development and validation of recombinant enzyme activity assays. These points include optimization of enzyme kinetic parameters, multi-dimensional evaluation strategies for complex mechanisms, precise control of product quantification, and approaches for developing reference materials. The aim is to provide a theoretical basis and technical support for the research, regulatory evaluation, and clinical safe application of innovative recombinant enzyme drugs.

Objective: To establish a GC-MS method for simultaneous determination of extractable amounts of 21 plasticizers from 4 common types (phthalates, citrate esters, terephthalates, polyester) in pharmaceutical packaging materials. Methods: Plasticizers in the pharmaceutical packaging material samples were extracted using n-hexane by ultrasound at 40 ℃ for 30 min. The extract was filtered through a 0.22 µm organic membrane filter, separated on a Select PAH capillary chromatography column, qualitatively analyzed by GC-MS in full scan mode, and quantitatively analyzed in selected ion monitoring mode. Results: All 21 plasticizers were completely separated within 35 min. In the concentration range of 0.02 to 2 µg · mL^-1, good linearity was observed between concentration and peak area, with correlation coefficients all above 0.994. The detection limits for all target compounds were less than 0.01 µg · mL^-1. The recoveries of spiked samples ranged from 87.8% to 118.0%. The method was applied to 14 batches of pharmaceutical packaging materials (plastic bottles, rubber stoppers, infusion bags), and plasticizers were detected in 6 batches, including diisobutyl phthalate (DIBP), di-n-butyl phthalate (DBP), bis (2-ethylhexyl) phthalate (DEHP), and acetyl tributyl citrate (ATBC). Conclusion: The method established in this study for the simultaneous determination of extractable amount of 21 plasticizers of different types has high sensitivity and good repeatability, and is suitable for rapid determination of plasticizers in pharmaceutical packaging materials.

Objective: To establish a high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for detection of the genotoxic impurity N-nitroso-desethyl-lidocaine in lidocaine hydrochloride and its injections. Methods: Chromatographic separation was achieved on a Capcell Pak C₁₈ column (150 mm×4.6 mm, 5 μm) through a gradient elution program with ammonium acetate solution-acetonitrile as the mobile phase at the flow rate of 1.0 mL · min^-1, the injection volume of 2 μL, and the column temperature of 30 ℃. A heated electrospray ionization (HESI) source was employed in the negative ion mode with selected ion monitoring (SIM) at m/z 234.3 for detection. Results: A good linear relationship was observed for N-nitroso-desethyl-lidocaine concentration in the range of 0.212-4.232 ng · mL^-1 with peak area (r=1.000). The limit of quantification (LOQ) and limit of detection (LOD) were determined to be 0.077 ng · mL^-1 and 0.025 ng · mL^-1, respectively. The mean recovery rates (n=9) were 101.2% (RSD=0.85%) for the active pharmaceutical ingredient and 99.1% (RSD=0.95%) for the injections. The impurity was detected in three batches of the raw material at levels ranging from 3 to 4 ng · g^-1, while it was not detected in any of the three injection batches. All results were within the acceptable limits. Conclusion: The developed method is highly sensitive and specific and can be employed for accurate detection of the genotoxic impurity N-nitroso-desethyl-lidocaine in the raw material and injections of lidocaine hydrochloride, thereby ensuring medication safety for patients.

In recent years, with the successive approval and market launch of multiple gene therapy products, this field has entered a phase of rapid development. This paper systematically reviews the research and development status as well as quality control of gene therapy products. Through an overview of the current development of gene therapy products and the progress in regulatory frameworks both domestically and internationally, this paper outlines the classification system and discusses the characteristics and clinical advancements of various types of products, including viral vectors, nucleic acid-based drugs, and oncolytic microorganisms. Regarding quality control, this paper expounds layer by layer from raw materials, in-process control, to lot release tests, summarizing common testing indicators and specific quality control considerations for different product types. Taking rAAV-based products as an example, it illustrates the framework of quality standards for viral vectors. Additionally, in light of clinical safety events related to critical quality attributes in recent years, this paper discusses the importance of enhancing quality control for clinical-stage investigational products, aiming to provide a systematic reference for the development and quality evaluation of gene therapy products.