31 May 2026, Volume 46 Issue 5
    

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    Review & Monography
  • Research progress in gel preparations and testing methods for their in vitro release and permeation*
    YU Yuan-fang, CHEN Hua, LI Jie, XU Hui, YU Li-ju
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 739-746. https://doi.org/10.16155/j.0254-1793.2025-0300
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    A gel system, a thick liquid or semisolid preparation with gel characteristics, is made of a drug substance and an excipient capable of forming a gel. Gel preparations are commonly used in the skin and body cavity, such as the nasal cavity, vagina, and rectum. Percutaneous administration of gel preparations can avoid the first-pass effect of the liver and improve the bioavailability of drugs. This paper summarized different kinds of gel preparations by reviewing the literature at home and abroad and expounded the in vitro diffusion and permeation related to the quality evaluation of gel preparations. The aim of this study is to provide reference for the research on gel preparations and the in vitro release test (IVRT) and in vitro permeation test (IVPT).
  • Name, origin and production areas research of Sinomenium acutum*
    TAO Ye, YU Kun-zi, CHENG Xian-long, LIU Shou-jin, LI Xiang-ri, ZHANG Ya-zhong, WEI Feng
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 747-753. https://doi.org/10.16155/j.0254-1793.2025-0565
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    Through a comprehensive review of classical and modern herbal literature, combined with field investigations, this study systematically examined the name, origin, and production areas of Qingfengteng (Chinese name). The results demonstrate that “Qingfengteng” was first officially documented as a medicinal name in Bencao Zeyao Gangmu. The plant described as “Qingfengteng” in Bencao Gangmu is conspecific with the modern medicinal plant bearing the same species. Analysis of the taxonomic history of its botanical nomenclature, along with data from specimen databases and field surveys, revealed that the pubescence characteristic of the variety Maoqingteng (hairy Qingfengteng) represents intraspecific variation—an ecologically adaptive phenotype rather than a stable classification and identification feature. Therefore, Maoqingteng should be classified within the species Sinomenium acutum (Fenglong, Chinese name), together with Qingfengteng. This study clarifies the historical evolution of the name and origins of Qingfengteng, thereby providing a scientific basis for its rational clinical use, resource identification, and quality control.
  • Modern research progress in the classical formula Yiguanjian*
    HOU Tian-yu, LIU Jing, LIN Yong-qiang, WEI Feng, HAN Na, WANG Qi
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 754-762. https://doi.org/10.16155/j.0254-1793.2026-0148
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    Yiguanjian, a classical formula for nourishing yin and soothing the liver, was first recorded in Xumingyileian · Xinweitongmen by Wei Zhi-xiu in the Qing Dynasty. It has been widely used in the clinical treatment of chronic liver diseases and related conditions. This paper systematically reviews the research progress in the chemical constituents, pharmacodynamic substances, network pharmacology, quality marker (Q-marker) prediction, and quality control methods of Yiguanjian. According to the Rule of Five (RO5) and the five principles of Q-markers, predictive analysis suggests that ferulic acid, ligustilide, psoralen, bergapten, and ruscogenin may serve as potential bioactive components and candidate Q-markers. Although substantial progress has been achieved in modern research on Yiguanjian, future studies should focus on establishing a Q-marker-based quality control system, integrating multiple analytical techniques to achieve whole-process quality control, and strengthening pharmacokinetic and dose-effect relationship studies to clarify compatibility mechanisms, thereby promoting the high-quality development and clinical application of this classical formula.
  • Identification and optimization strategy for critical parameters during development process of peptide mapping analysis methods for antibody drugs
    NI Yong-bo, GUO Lu-yun, CUI Yong-fei, CHEN Hao-bin, CAI Zhen, YU Chuan-fei
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 763-773. https://doi.org/10.16155/j.0254-1793.2025-0482
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    Antibody drug is an important component in the biopharmaceutical industry, so its quality control is critical. Peptide mapping analysis is a key technique for characterizing the structure and quality of antibody drugs. Through the separation and detection of peptides after enzymatic digestion of antibodies, critical information such as amino acid sequences and post-translational modifications can be provided. This paper described the critical parameters in the development of peptide mapping analysis methods for antibody drugs, including sample pretreatment, enzymatic digestion conditions, chromatographic separation, and mass spectrometric detection. The control strategies for these parameters were also discussed, aiming to provide a reference for establishing efficient, accurate, and reliable peptide mapping analysis methods for antibody drugs to satisfy the requirements for antibody drug research and development, production, and quality control. By analyzing the core parameters and control logic in the development of peptide mapping analysis methods for antibody drugs, a full-process scheme covering sample pretreatment, enzymatic digestion, separation, and detection was established. This paper provides technical references for biopharmaceutical enterprises and regulatory authorities to establish efficient, accurate, and reliable quality control systems for antibody drugs, helping to improve the research and development efficiency and production quality of antibody drugs and ensuring the safety and efficacy of clinical medications.
    • Ingredient Analysis
  • A block-based molecular networking strategy for qualitative analysis of chemical constituents in the herb pair Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos*
    ZHAO Qing-rong, LAN Ting-ting, YU Dai-xin, WU Qi-nan, QU Cheng
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 774-800. https://doi.org/10.16155/j.0254-1793.2025-0535
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    Objective: To rapidly analyze flavonoids, diterpenoids, and phenolic acids in the classic herb pair Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos (SM-CF) by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q TOF MS) combined with block-based molecular networking (BBMN). Methods: UHPLC-Q TOF MS was employed to acquire raw MS data from the herb pair. Gradient elution was achieved through an ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) at a column temperature of 35 ℃, with 0.1% formic acid in water as mobile phase A and acetonitrile as mobile phase B at a flow rate of 0.3 mL · min-1. An electrospray ionization (ESI) source was used for scanning in both positive and negative ion modes over the m/z range 100-1 500. BBMN was constructed via the Global Natural Products Social Molecular Networking (GNPS) platform. Characteristic biosynthetic fragments of flavonoids, diterpenoids, and phenolic acids were efficiently identified through the BBMN. Cytoscape 3.10.2 was used to visualize molecular clusters. The major chemical components in the herb pair were annotated by combining information from the GNPS platform, reference standards, literature reports, and database matching. Results: Compared with feature-based molecular networking (FBMN), BBMN significantly streamlined data processing by screening with characteristic biosynthetic fragments (increasing the node hit rate for flavonoids by 20 folds from 0.99% to 20.14%). Through the BBMN strategy, 118 components were annotated in the herb pair SM-CF, including 62 flavonoids, 33 diterpenoids, and 23 phenolic acids. Among them, 21 compounds were identified for the first time in this herb pair. Additionally, 42 chemical constituents were further identified by conventional methods. Conclusion: The BBMN strategy greatly improves the efficiency of MS data interpretation by recognizing characteristic biosynthetic fragments, enabling highly selective screening of target compounds and outperforming conventional FBMN. This approach allows rapid and accurate identification of flavonoids, diterpenoids, and phenolic acids in SM-CF, providing a valuable reference for quality control of this herb pair and the rapid characterization of chemical components in complex traditional Chinese medicine systems.
  • A novel pretreatment technique based on vortex-salting out-solid-phase extraction for the determination of triptolide content in Jinguan tablets*
    GUO Lin, ZHANG Qing, GUO Sheng-he, DAI Shu-wen, JIN Hong-yu, WEI Feng, WANG Ya-dan
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 801-807. https://doi.org/10.16155/j.0254-1793.2025-0543
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    Objective: To establish an accurate and efficient HPLC method for the determination of triptolide content in Jinguan tablets, addressing the complex pretreatment, time-consuming process, and poor purification effect of the current standard method, and thus provide technical support for improving the quality standard of this product. Methods: The test solution was prepared with acetonitrile-water as the extraction solvent, followed by vortex oscillation extraction, salting-out, and purification with a neutral alumina solid-phase extraction (SPE) column. HPLC separation was performed through a Halo 90Å C18 column (250 mm×4.6 mm, 5 μm) with the column temperature maintained at 30 ℃. Methanol-water (35 ∶ 65, v/v) was used as the mobile phase, at a flow rate of 1.0 mL · min-1. The detection wavelength was set at 220 nm. Results: The triptolide concentration in the range of 1.106 6-44.262 2 μg · mL-1 showed a good linear relationship with peak area (r=1.000). The relative standard deviation (RSD) values (n=6) in precision, stability, and repeatability tests were all less than 2.3%. The average recovery rates (n=3) at high, medium, and low addition concentration levels were 108.5%, 95.4%, and 98.0%, with RSD values of 1.6%, 2.9%, and 1.8%, respectively. The method demonstrated good robustness in the terms of SPE column and chromatographic column. The triptolide content in nine batches of Jinguan tablets ranged from 5.00 to 7.03 μg per tablet. Taking three batches as examples, their determination results were significantly higher than those obtained by the current standard method, suggesting that the method established in this study had higher extraction efficiency for triptolide. Conclusion: The established method for determining triptolide content is sensitive, accurate, efficient, and environmentally friendly. On the basis of accumulating more batch determination data and revising the content limit, this method is expected to replace the current standard method for quality control of Jinguan tablets and provide a reference for quality control of other preparations containing Tripterygii Radix et Rhizoma.
  • Research on dynamic change laws of carbohydrate components in Fritillaria unibracteata Hsiao et K. C. Hsia with different growth years*
    ZHENG Han-xiao, LI Meng-yu, PU Jing-zhe, HU Chong, CHEN Ling-li, ZHANG Ya-zhong
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 808-818. https://doi.org/10.16155/j.0254-1793.2025-0447
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    Objective: To investigate the dynamic changes of carbohydrate components in Fritillaria unibracteata Hsiao et K. C. Hsia at different growth years, and to provide a scientific basis for standardized cultivation, harvesting, and quality control. Methods: The polysaccharide content of F. unibracteata was quantified using the sulfuric acid-anthrone method, with detection at 625 nm. The monosaccharide composition and content were determined by HPLC-UV following acid hydrolysis and pre-column derivatization with PMP. Separation was carried out on a Waters Symmetry C18 column (250 mm×4.6 mm, 5 μm) at 35 ℃, with a mobile phase consisting of acetonitrile and 0.02 mol · L-1 ammonium acetate (17 ∶ 83, v/v) at a flow rate of 1 mL · min-1. Detection was performed at 245 nm, and the injection volume was 5 μL. In addition, sucrose and starch contents were analyzed by HPLC-ELSD using an Alltech Prevail Carbohydrate ES column (250 mm×4.6 mm, 5 μm) maintained at 35 ℃. The mobile phase was acetonitril-water (75 ∶ 25, v/v) at a flow rate of 1 mL · min-1. The drift tube temperature was set to 110 ℃, with a gas flow rate of 2.5 L · min-1 and an injection volume of 5 μL. Results: Significant differences in carbohydrate contents were observed in F. unibracteata at different growth years. The mean polysaccharide content showed a dynamic trend of increase, decrease, and subsequent increase, with values of 5.46 mg · g-1, 6.04 mg · g-1, 5.14 mg · g-1, and 8.10 mg · g-1 in two-year-grown, three-year-grown, four-year-grown, and five-year-grown samples, respectively. Glucose was the predominant monosaccharide in polysaccharide hydrolysates, and its proportion increased significantly with increasing growth years, ranging from 58.84% to 65.77%. Rhamnose, galactose, and arabinose were present at relatively low levels without a clear trend. The mean sucrose content exhibited dynamic accumulation characteristics, increasing from 54.54 mg · g-1 in two-year-grown samples to 64.28 mg · g-1 in three-year-grown samples, slightly decreasing to 58.59 mg · g-1 in four-year-grown samples, and then rapidly increasing to 97.60 mg · g-1 in five-year-grown samples. The mean starch content increased annually from two to four years of growth (178.42-233.51 mg · g-1), reached a peak at four years, and then slightly decreased to 216.55 mg · g-1 in five-year-grown samples. Conclusion: The carbohydrate contents in F. unibracteata differ significantly among different growth years and exhibit regular variation patterns with prolonged growth duration. Polysaccharide and sucrose contents are highest in five-year-grown bulbs, whereas starch accumulation peaks in four-year-grown bulbs. Glucose is the major monosaccharide and increases with growth years. Sucrose, starch, and polysaccharides are suggested as important indicators for the quality evaluation of F. unibracteata, providing a scientific basis for optimizing harvest time and quality control.
    • Metabolism Analysis
  • Determination of seven vitamin D and metabolites in human serum based on HPLC-MS/MS*
    LI Pei, PAN Jing-jue, ZHAO Guo-fang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 819-829. https://doi.org/10.16155/j.0254-1793.2025-0437
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    Objective: To establish a liquid-liquid extraction (LLE) combined with derivatization and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for accurate quantification of seven vitamin D and metabolites in the human serum. Methods: Methanol-isopropanol (8 ∶ 2, containing 0.3% formic acid) was used as a protein denaturation reagent, and n-hexane-ethyl acetate (7 ∶ 3) as the extraction solvent. Derivatization was carried out with 25 μg of 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD), with the reaction reaching stability after 1 h. Chromatographic separation was performed through a Waters Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) by gradient elution with the mobile phase consisting of water and methanol, each containing 0.1% formic acid and 2.5 mmol · L-1 ammonium formate, at a flow rate of 0.3 mL · min-1. The column temperature was set at 40 ℃. Detection was carried out via electrospray ionization in the positive mode (ESI+) with multiple reaction monitoring (MRM). Results: At three spiked concentration levels (2, 20, and 200 μg · L-1), the recovery rates of seven vitamin D and metabolites (vitamin D2, vitamin D3, 25-hydroxy vitamin D2, 25-hydroxy vitamin D3, 1,25-dihydroxy vitamin D2, 1,25-dihydroxy vitamin D3, and 24R, 25-dihydroxy vitamin D3) ranged from (71.3±3.5)% to (115.9±10.6)%. The linear correlation coefficients for these analytes ranged from 0.998 4 to 0.998 8 within the concentration range of 0.25-80 μg · L-1. The method limits of detection (LODs) and quantification (LOQs) were 0.022-0.10 μg · L-1 and 0.073-0.34 μg · L-1, respectively. The intra-day and inter-day relative standard deviations (RSDs) were 4.1%-11.3% and 6.5%-15.0%, respectively. This method was then used to analyze the seven compounds in 30 serum samples from individuals aged 60 years and above. All the seven compounds were detected, with 25-hydroxy vitamin D3, 1,25-dihydroxy vitamin D3, and 24R,25-dihydroxy vitamin D3 being the primary metabolites, exhibiting median concentrations of 4.30, 2.37, and 0.57 μg · L-1, respectively. Conclusion: This method is fast and high-throughput, enabling simultaneous quantitation of seven vitamin D and metabolites in the serum within approximately 5 h. It demonstrates high accuracy, excellent sensitivity, and good stability, with the LOD about one order of magnitude lower than that reported in the literature.
  • Determination of the plasma concentration of IMM in Beagle dogs by high performance liquid chromatography-tandem mass spectrometry*
    ZHANG Kun, SHAO Hui-hui, HAN Zun-sheng, ZHANG Chi, LIU Bo, ZHANG Jie, LIU Chun-xi, WU Song, YANG Qing-yun
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 830-837. https://doi.org/10.16155/j.0254-1793.2025-0411
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    Objective: To develop a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determining the plasma concentration of methyl 7,7’-dimethoxy-5’-(morpholinomethyl)-[4,4’-bibenzo[d][1,3] dioxole]-5-carboxylate, methanesulfonate (IMM) in Beagle dogs. Methods: Plasma samples were treated by the protein precipitation method and analyzed by HPLC-MS/MS. An Agilent Poroshell 120 EC-C18 column (50 mm×2.1 mm, 2.7 μm) was used for gradient elution with 0.1% formic acid in water as the mobile phase A and 0.1% formic acid in acetonitrile as the mobile phase B. A gradient elution program was carried out with a flow rate of 0.4 mL · min-1, a column temperature at 35 ℃, and a injection volume of 1 μL. MS detection was performed with electrospray ionization (ESI+) as the ion source under the multiple reaction monitoring (MRM) mode. The qualified ion pairs were m/z 460.1→341.0 (IMM) and m/z 237.0→194.0 (carbamazepine, internal standard). Other MS parameters were a drying gas temperature of 150 ℃, a drying gas flow rate of 11 mL · min-1, a sheath gas temperature of 300 ℃, a sheath gas flow rate of 11 mL · min-1, and a capillary voltage of 3 000 V. Results: The calibration curve was in linearity (r=0.999 9) within the plasma concentration range of 5-8 000 ng · mL-1 for IMM. The lower limit of quantification was 5 ng · mL-1. The average extraction recovery varied within the range of 95.3%-110.5%, with RSD≤6.3% (n=6). The matrix effect varied within the range of 101.8%-108.8%, with RSD≤10.9% (n=6). The accuracy values were 89.4%-105.6% and the precision (RSD) values were≤7.4% (intra-day, n=6; inter-day, n=30). The plasma samples remained stable in the injectors (4 ℃) for 24 h, at room temperature for 24 h, after 3 freeze-thaw cycles, and at -20 ℃ for 30 d. The IMM API and IMM tablets in Beagle dogs showed the AUC0-24 h values of (9 601.45±1 912.26) μg · h · L-1 and (8 758.39±1 785.31) μg · h · L-1, Cmax values of (3 103.05±718.38) μg · L-1 and (1 274.18±338.03) μg · L-1, and t1/2 values of (5.75±1.36) h and (6.13±2.15) h, respectively. The relative bioavailability of IMM tablets was determined to be 95.14%. The pharmacokinetic results indicated that the tablets had a certain sustained-release effect. Conclusion: This method is highly specific, sensitive, accurate in results, and simple to operate. It can be used for the determination of plasma concentrations of IMM API and IMM tablets in Beagle dogs after administration. This method provides reliable technical reference and a scientific basis for preclinical studies of the new drug IMM and its tablets.
  • Identification of components absorbed into serum after administration of Bufei Jianpi granules based on UPLC-MS/MS*
    XU Han-han, LI Rong-rong, SUN Shu-ding, ZHENG Li-shi, ZHAO Di, FENG Su-xiang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 838-850. https://doi.org/10.16155/j.0254-1793.2025-0408
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    Objective: To analyze and identify the components absorbed into the serum after administration of Bufei Jianpi granules by UPLC-MS/MS. Methods: A Hypersil GOLD column (100 mm×2.1 mm, 2.6 μm) was used with a column temperature of 30 ℃. Gradient elution was conducted with the mobile phase composed of methanol (A) and 0.1% formic acid aqueous solution (B) at a flow rate of 0.2 mL · min-1. Electrospray ionization (ESI) was adopted, and full MS/dd-MS2 scanning was performed in positive and negative ion modes for data-dependent acquisition (DDA). The absorbed components were analyzed by comparison with the data of reference substances, literature, and databases. Results: After rats were administrated with Bufei Jianpi granules, 50 prototype compounds and 52 metabolites were identified in the serum, primarily including flavonoids, phenylpropanoids, alkaloids, and terpenoids. The metabolic pathways included phaseⅠmetabolism such as demethylation, oxidation, and reduction, and phaseⅡmetabolism such as glucuronidation and sulfation. Conclusion: This study analyzes the components absorbed into the serum after administration of Bufei Jianpi granules, providing a scientific basis for revealing the material basis and mechanism of Bufei Jianpi granules in the treatment of chronic obstructive pulmonary disease.
    • Bioassay
  • Tissue distribution of mesenchymal stem cells in rats after single injection into knee joint cavity*
    TAN Jin-xiu, XIAO Hai-rong, ZHANG Chen, LI Wei, HU Jin-fang, YAN Feng-ying
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 851-860. https://doi.org/10.16155/j.0254-1793.2025-0316
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    Objective: To establish and validate a method for detecting human-derived mesenchymal stem cells (MSCs) in rats, and to study their tissue distribution in immunodeficient F344RG rats after injection into the knee joint cavity. Methods: A real-time quantitative polymerase chain reaction (qPCR) method targeting a specific sequence on human chromosome 20 was established for the detection of human-derived MSCs in rats, with a lower limit of quantification of 0.015 36 ng per reaction. The method was fully validated. The qPCR mixture (25 μL)contained 12.5 μL Premix Ex TaqTM (Probe qPCR), 0.5 μL each of primers and probe (1×10-5 mol · L-1), 5 μL template, and nuclease-free water. The cycling protocol was 95 ℃ for 30 s (1 cycle) and then 45 cycles of 95 ℃ for 5 s and 54 ℃ for 30 s, with fluorescence signal acquired during the annealing/extension step. F344RG rats received a single injection of human-derived MSCs into the knee joint cavity. Whole blood, heart, liver, spleen, lung, kidney, brain, bone marrow, gonads (testes/ovaries), and joint tissue from the injection site were collected at 1 d, 3 d, 5 d, 7 d, 14 d, 28 d and 42 d post-dose. Genomic DNA was extracted, and the presence of MSCs in blood and tissue samples was determined by qPCR. Results: A qPCR method for the detection of human-derived MSCs in whole blood and various tissue samples of F344RG rats was successfully established, which was not affected by the gender of the MSCs donor. MSCs were detected in the local joint tissue of all the rats within 42 d after administration, with mean content gradually decreasing from 297.83 ‱ to 1.21 ‱. MSCs were also detected in the lungs of 1 out of 6 rats at 5 d and 7 d post-dose, with the content of 1.99 ‱ and 0.94 ‱, respectively. No MSCs were detected in any other tested tissue or organ samples. Conclusion: The established qPCR method is suitable for the detection of human-derived MSCs in rats. After a single injection into the knee joint cavity of F344RG rats, MSCs are mainly distributed in the joint tissue, and their content gradually decreases over time. Additionally, MSCs can enter the lungs in individual rats (2 out of 42 rats), with the precise mechanism remaining to be deciphered.
  • Comprehensive structural characterization of lecanemab based on liquid chromatography-high-resolution mass spectrometry*
    XU Jun-wei, FU Jian-ping, XU Zhi-yong, LIU Yun-fei, KE Chun-shan, YUAN Yang-yang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 861-870. https://doi.org/10.16155/j.0254-1793.2025-0404
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    Objective: To comprehensively characterize the structure of lecanemab based on liquid chromatography (LC) coupled with high-resolution Orbitrap mass spectrometry (HRMS). Methods: This study employed reversed-phase chromatography (PLRP-S column, 50 mm×2.1 mm, 5 μm, 1 000 Å) coupled with HRMS for structural characterization of lecanemab. The column temperature was set at 60 ℃, and gradient elution was performed with the mobile phase composed of 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B) at a flow rate of 0.3 mL · min-1. Firstly, intact proteins and their subunits were analyzed via MS1 in the positive ion mode to determine their molecular mass, followed by analysis of reduced samples to determine light and heavy chain molecular mass. After digestion with trypsin and chymotrypsin, peptides were separated through a C18 column (50 mm×2.1 mm, 2.6 μm). Data-dependent acquisition was employed for peptide sequencing and disulfide bond identification, while MS/MS was employed to characterize the post-translational modifications (PTMs) including oxidation, deamidation, and glycosylation. Results: The relative molecular mass of intact lecanemab was determined to be 149 977, with the mass of 146 926 following N-glycan removal. Complete sequence coverage (100%) was achieved for both the light chain and heavy chain, and all the 16 disulfide bonds were accurately identified, consistent with theoretical expectations. Key PTMs identified included oxidation, N-terminal lysine loss, deamidation, and N-glycosylation, with predominant glycoforms such as G0F, G1F, and G2F being detected. Conclusion: This study successfully achieves comprehensive structural characterization of lecanemab through reversed-phase chromatography coupled with HRMS, including intact relative molecular mass determination, subunit analysis, peptide sequence coverage, and PTM identification. The methodology provides technical reference for structural characterization of similar monoclonal antibodies.
    • Quality Control
  • DNA degradation patterns and molecular detection methods in the processing of tortoise-shell glue and deer-horn glue*
    ZHANG Quan-fang, LUO Li-yun, BI Wu, DONG Shu-guang, BU Xun, LIN Yong-qiang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 871-884. https://doi.org/10.16155/j.0254-1793.2025-0192
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    Objective: To investigate the degradation patterns of genomic DNA during the key processing stages of tortoise-shell glue (Mauremydis Carapacis et Plastri Colla) and deer-horn glue (Cervi Cornus Colla), and to establish a molecular identification method for authentic origin based on DNA barcoding and nested real-time fluorescence quantitative PCR. Methods: Samples were collected at the raw material, pre-concentration, post-concentration, glue solution, and finished product stages during the processing of tortoise-shell glue and deer-horn glue. The whole-genome DNA extraction method was optimized, and the quality and degradation of DNA were assessed by spectrophotometry and agarose gel electrophoresis. DNA barcoding identification was performed using universal animal primers for the mitochondrial CO I or Cytb genes. Two pairs of specific primers and one probe were designed targeting the mitochondrial 16S rRNA sequences of turtle and deer, and an exogenous internal control was introduced to monitor the PCR amplification system. A combined detection method based on DNA barcoding and nested real-time fluorescence quantitative PCR was established to ensure the accuracy of samples throughout the entire processing procedure. Results: The highest DNA purity in tortoise-shell glue was observed in the post-concentration and glue juice stages, while higher DNA content was found at the cleaned turtle shell and pre-concentration stages. During deer-horn glue processing, the A260/A280 ratios of DNA were consistently below 1.7, with the highest DNA content in the glue juice stage. Agarose gel electrophoresis showed clear single bands at the raw material stage for both types of glue, while varying degrees of smearing appeared in the subsequent processing stages. DNA barcoding technology enabled accurate identification of raw and cleaned raw materials. Nested real-time fluorescence quantitative PCR allowed detection throughout the entire process with high sensitivity, achieving limits of detection of 0.16 ng for raw materials and 4 ng for finished glue products. Conclusion: Genomic DNA undergoes progressive degradation as processing proceeds during the manufacture of tortoise-shell glue and deer-horn glue. The combination of the developed nested real-time fluorescence quantitative PCR method and DNA barcoding technology is rapid and accurate, meeting the requirements for DNA traceability at all processing stages and providing reliable technical support for the origin identification of these two types of glue medicinal materials.
  • Construction of a quality-grade scorecard system for Carthami Flos based on test items in the Chinese Pharmacopoeia and its quantitative evaluation of the standards*
    WANG Xiao-wei, GENG Yi-wei, SHI Yan, LI Gui-ben, WANG Hai-bo, LI Yan-chao, LI Yi-xian, YANG Yuan, YIN Kun-kun
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 885-896. https://doi.org/10.16155/j.0254-1793.2025-0735
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    Objective: To develop a scorecard system that uses the test items stipulated in the Chinese Pharmacopoeia as scoring criteria for identifying the quality grades of Carthami Flos (Honghua). Methods: A total of 296 batches of Carthami Flos samples were first classified into four sensory grades (A, B, C, and D) by experienced evaluators. All the samples were then tested according to the Chinese Pharmacopoeia, yielding data for eight items: moisture, total ash, acid-insoluble ash, absorbance, content of hydroxysafflor yellow A, content of kaempferol, content of extract, and content of foreign matter. The 296 batches were randomly split into training and test sets. Training-set data were binned, and the weight of evidence (WOE) and information value (IV) of each bin were calculated. Bins were merged and optimized until the best segmentation was reached. After WOE transformation, a logistic regression model was built and a scoring scheme was designed. With 50 points as the score where the odds of positive vs. negative samples equal 1, the base score and the score for every bin were derived. The final score was the sum of the base score and the bin score corresponding to the original test result, and 50 points served as the threshold for grade discrimination. Three parallel scorecards (Card-Ⅰ, Card-Ⅱ, and Card-Ⅲ) were constructed. The importance of each pharmacopoeial item was quantified from its bin scores under the relevant card. Results: Card-Ⅰ, Card-Ⅱ, and Card-Ⅲ enabled the discrimination of Carthami Flos samples between grade A and non-Grade A, between grades A&B and grades C&D, and between non-grade D and grade D, respectively. Card-Ⅰ, Card-Ⅱ, and Card-Ⅲ achieved the discrimination accuracy of 86.67%, 80.00%, and 73.33%, respectively, on the test set. Conclusion: The three scorecards effectively bridge the objective pharmacopoeial standards and the subjective sensory grading of Carthami Flos, enabling reliable quality-grade discrimination and quantitative evaluation of individual standard items according to practical testing needs. Combining conventional herbal analysis with data science and artificial intelligence allows objective analytical indicators to replace subjective sensory judgement and makes the performance of each indicator quantifiable.
  • Comprehensive quality evaluation of Tetrastigma hemsleyanum based on appearance characteristics and internal components*
    PAN Ping, LUO Yi-yuan, CAI Zhong-qi, JIANG Yu-chen, FU Jia-yi, HU Sun-xian, CHEN Hong-jiang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 897-910. https://doi.org/10.16155/j.0254-1793.2025-0329
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    Objective: To explore the correlation between the appearance characteristics and internal component content of Tetrastigma hemsleyanum and to provide a theoretical basis for establishing an objective and scientific comprehensive quality evaluation method for this medicinal material. Methods: The single-grain weight, length, diameter, cross-sectional color value (L*a*b*), and ethanol-soluble extract content of T. hemsleyanum were determined using an electronic balance, a digital vernier caliper, and a colorimeter, respectively. The content of total flavonoids, total polyphenols, and total polysaccharides was determined by UV-visible spectrophotometry at detection wavelengths of 510, 490, and 756 nm, respectively. The content of 4 phenolic acids, 13 flavonoids, 12 nucleosides, and 12 amino acids in T. hemsleyanum were analyzed by UPLC-MS/MS with multiple reaction monitoring (MRM). Pearson correlation analysis was applied to investigate the correlation between appearance characteristics and internal components, and the TOPSIS method was employed for the comprehensive quality evaluation of the medicinal material. For the analysis of flavonoids and phenolic acids, separation was performed on a Waters BEH Shield RP C18 column with a mobile phase consisting of 0.1% formic acid in water and acetonitrile under gradient elution, with ESI- mode. For the analysis of nucleosides and amino acids, separation was performed on a Waters XBridge Amide column with a mobile phase consisting of 0.2% formic acid in water and 0.2% formic acid in acetonitrile under gradient elution, with ESI+ mode. The common mass spectrometry parameters were as follows: capillary voltage 3.0 kV, ion source temperature 150 ℃, desolvation gas flow rate 800 L · h-1, cone gas flow rate 50 L · h-1, and desolvation gas temperature 550 ℃. Results: For the 23 batches of T. hemsleyanum samples, the single grain weight ranged from (0.74±0.81) g to (17.24±9.94) g, the length ranged from (16.94±6.20) mm to (51.37±13.28) mm, the diameter ranged from (9.29±2.24) mm to (24.66±4.65) mm, the cross-section L* value ranged from (60.08±5.10) to (81.81±7.74), a* value ranged from (4.45±1.64) to (12.36±2.72), b* value ranged from (14.19±5.21) to (24.36±3.54), and the ethanol-soluble extract content was 6.51% to 16.27%, indicating significant differences in morphological and color parameters among the samples. The content of total flavonoids, total polyphenols and total polysaccharides were 7.54-28.48 mg · g-1, 2.19-11.22 mg · g-1, and 1.69-34.26 mg · g-1, respectively, with the variation in total polysaccharide content being the most pronounced. In addition, a total of 41 components, comprising 4 phenolic acids, 13 flavonoids, 12 nucleosides, and 12 amino acids in T. hemsleyanum, were showed remarkable differences among different batches. The phenolic acids were mainly represented by gallic acid and neochlorogenic acid, while catechin and kaempferol-3-O-rutinoside exhibited relatively prominent content among the flavonoids; nucleosides and amino acids also showed marked inter-batch differentiation. Pearson correlation analysis demonstrated that single-grain weight and diameter were significantly positively correlated with the content of total flavonoids and total phenolic acids (P<0.01), and could characterize the content of flavonoids, phenolic acids, and polysaccharides. A significant correlation was observed between appearance characteristics and internal component content. The TOPSIS comprehensive evaluation revealed that the highest comprehensive score (Ci) was obtained for the sample S3 from Yueqing, Wenzhou, Zhejiang. Conclusion: The results of this study lay an experimental foundation for the establishment of a comprehensive quality evaluation system for T. hemsleyanum and provide a scientific basis for the screening of superior germplasm resources of this medicinal material.
    • Standard Deliberation·Rapid Analysis
  • A method for determining the encapsulation efficiency of hydrophobic drug AST2169-loaded liposomes
    LI Lu, ZHANG Xue, LIANG Chun-qing, JIANG Wen-ming, QIAN Min, JIN Wei, LE Jian
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 911-918. https://doi.org/10.16155/j.0254-1793.2025-0534
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    Objective: To establish a method for determining the encapsulation efficiency of liposomes loaded with hydrophobic drugs, addressing the limitations of conventional gel column chromatography in separating liposomes from hydrophobic free drugs. Methods: A modified NW Rose 6FF agarose gel column (1 cm×3.6 cm) was used. After precise loading of 100 μL test sample stock solution, 0.9% sodium chloride solution and 70% ethanol solution (10 mL each) were successively loaded for gradient elution, and encapsulated and free drug fractions were collected. A Waters Xbridge Shield RP18 column with the column temperature of 40 ℃ was used for gradient elution, which was carried out with the mobile phase consisting of acetonitrile and 5 mmol · L-1 ammonium acetate solution (90 ∶ 10) at a flow rate of 1 mL · min-1. The detection wavelength was set at 228 nm, and the injection volume was 5 μL. The concentration of each fraction was measured by HPLC-UV, and the encapsulation efficiency was finally calculated. Results: The optimized method achieved efficient separation between liposomes and free drugs. The precision (relative standard deviation, RSD) of encapsulation efficiency determination was 1.0%, and the average recovery rate was 100.0%. The mean encapsulation efficiency determined was 92.8%, and the column recovery rates were all above 95% (97.8%-102.4%). Conclusion: The improved agarose gel column chromatography method is mild and efficient, and suitable for determining the encapsulation efficiency of hydrophobic drug-loaded liposomes, providing a reliable tool for their quality control.
  • A rapid identification method of components derived from Cervus nippon based on loop-mediated isothermal amplification*
    ZOU He-yuan, GU Li-fei, WANG Hong, ZHANG Wen-xian, WANG Bing, WANG Shu-hong, SU Chang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 919-925. https://doi.org/10.16155/j.0254-1793.2025-0020
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    Objective: To establish a loop-mediated isothermal amplification (LAMP) method for identifying Cervus nippon, enabling visual and rapid identification of C. nippon-derived components. Methods: An online tool was employed to design a specific LAMP primer set for C. nippon based on the mitochondrial whole genome sequences of C. nippon, C. elaphus, and Rangifer tarandus. Then, the reaction temperature was optimized, and the sensitivity and specificity of the system were evaluated. The turbidity method and dye method were used to verify the method by comparison. At last, after incubation at the optimal temperature of 61 ℃ for 60 min, the dye method was adopted to test the reproducibility of LAMP on a large batch of samples identified based on COⅠ barcodes. Results: The LAMP method exhibited strong specificity and high sensitivity (0.324 5 pg · μL-1). Only components derived from C. nippon showed changes in turbidity curve and color responses, appearing blue. Other samples showed no amplification reaction and remained colorless. Conclusion: The constructed LAMP method for identifying C. nippon provides strong technical support for on-site rapid inspection and market supervision of C. nippon-derived products.
    • Safety Monitoring
  • Detection of related substances of caspofungin acetate and its preparation*
    LIU Hai-tao, KOU Jin-ping, HOU Jin-feng, LIU Qi, YU Li, CHE Bao-quan
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 926-939. https://doi.org/10.16155/j.0254-1793.2025-0162
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    Objective: To establish a detection method for related substances in caspofungin acetate and its preparation, and to study its impurity profile. Methods: Gradient elution was carried out through a Waters Symmetry C18 column (250 mm×4.6 mm, 5 μm) at the column temperature of 45 ℃, with NaCl-HClO4 solution as mobile phase A and acetonitrile as mobile phase B. The detection wavelength was set as 220 nm. Results: There were 25 potential related substances with known structures in caspofungin acetate. The proposed method showed good separation of related substances, with a limit of detection being 0.05 μg · mL-1 (equivalent to 0.003% of the concentration of the test solution) and a good linear relationship. The test solution was basically stable within 8 h at 5 ℃. Conclusion: This method can effectively separate and detect related substances of caspofungin acetate, with high sensitivity, accuracy, and repeatability.
  • Detection of N-nitroso-desmethyl-chlorpheniramine in chlorpheniramine maleate by UPLC-MS/MS*
    WANG Chen-yu, BAO Yu-zhang, YUAN Song, DI Bin, LIU Yang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 940-946. https://doi.org/10.16155/j.0254-1793.2025-0686
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    Objective: To establish an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the detection of the genotoxic impurity N-nitroso-desmethyl-chlorpheniramine in chlorpheniramine maleate. Methods: A Shim-pack Velox Biphenyl column (150 mm×3.0 mm, 2.7 μm) was employed, the column temperature was maintained at 40 ℃, with a mobile phase consisting of 10 mmol · L-1 ammonium formate containing 0.1% formic acid (A) and acetonitrile (B) under gradient elution at a flow rate of 0.3 mL · min-1. Detection was carried out by electrospray ionization (ESI) in the positive ion mode with multiple reaction monitoring (MRM) for the quantification of N-nitroso-desmethyl-chlorpheniramine. Results: N-nitroso-desmethyl-chlorpheniramine showed a good linearity over the concentration range of 0.003-1.01 ng · mL-1, with a correlation coefficient r>0.99. The spiked recoveries of N-nitroso-desmethyl-chlorpheniramine in the active pharmaceutical ingredient (n=3) ranged from 87.7% to 93.1%, with RSDs ranging from 0.39% to 2.9%. The recoveries (n=3) at low, medium, and high spiked concentrations in the finished product ranged from 90.3% to 92.9%, with RSDs ranging from 0.36% to 2.2%. The limit of detection was 0.000 7 ng · mL-1, and the limit of quantification was 0.002 ng · mL-1. Based on chlorpheniramine maleate, the content of N-nitroso-desmethyl-chlorpheniramine in the API was 10.41 ng · g-1, while the content in tablets from four manufacturers was 10.56 ng · g-1 (manufacturer A), 13.57 ng · g-1 (manufacturer B), 13.72 ng · g-1 (manufacturer C) and 3.43 ng · g-1 (manufacturer D). Conclusion: The established method is highly sensitive and specific, and can be used for the detection of the N-nitroso-desmethyl-chlorpheniramine impurity in chlorpheniramine maleate API and finished products, providing a reference for the quality control of chlorpheniramine maleate.
  • Detection and safety evaluation of 146 kinds of veterinary drug residues in imported Gekko gecko Linnaeus based on UPLC-MS/MS veterinary drug database platform
    WANG Jing, LIU Guan-ke, CHENG Ning-ning, ZHENG Xin-yuan, ZHOU Jun, YANG Tao
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(5): 947-956. https://doi.org/10.16155/j.0254-1793.2025-0789
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    Objective: To perform the detection and safety evaluation of 146 veterinary drug residues in imported Gekko gecko Linnaeus based on the UPLC-MS/MS veterinary drug database platform. Methods: The samples were extracted with 50% methanol and purified via n-hexane extraction. Separation was conducted through a Shim-pack GIST-HP C18-AQ column (100 mm×2.1 mm, 1.9 μm) by gradient elution with 2 mmol · L-1 ammonium acetate (containing 0.2% formic acid) and methanol (containing 0.2% formic acid) as the mobile phases at a flow rate of 0.4 mL · min-1 and a column temperature of 40 ℃. Veterinary drug residues were detected by tandem mass spectrometry with a positive electrospray ionization (ESI+) source under the multiple reaction monitoring (MRM) mode. Quantitation was performed with the matrix-matched external standard method. Results: Good linear relationships were observed for all the 146 veterinary drugs within a specific mass concentration range (all r>0.99). The limits of detection (LODs) of the method were determined to be in the range of 4.950-5.605 μg · kg-1, and the limits of quantitation (LOQs) were in the range of 24.750-28.025 μg · kg-1. The relative standard deviations (RSDs) for precision were in the range of 0.65%-8.27%. At three spiking concentration levels, the recoveries were in the range of 71.3%-138.4%, with corresponding RSDs ranging from 0.21% to 24.50%. In the screening of 25 batches of imported Gekko gecko Linnaeus, testosterone, 1,7-dimethylxanthine, and acetaminophen showed high detection rates. Conclusion: The construction of a veterinary drug database platform can effectively fill gaps in the detection of veterinary drug residues in the pharmaceutical field. The established screening technology is simple, fast, economical, and practical and can be used to simultaneously determine multiple veterinary drug residues in Gekko gecko Linnaeus.
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