31 July 2026, Volume 46 Issue 7
    

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    Review & Monography
  • Research progress of abrine in the field of pharmacy*
    LIN Quan, WEN Zi-shuai, XIAO Rui, WEI Ya-nan, FU Lei, ZHANG Chen-rui, CHEN Hai-ping, HUANG Xiao-lan, YAO Chun, WU Fang-fang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1169-1179. https://doi.org/10.16155/j.0254-1793.2025-0656
    Abstract ( )   Knowledge map   Save
    Abrine is an alkaloid derived from plants of the genus Abrus (Fabaceae), including A. cantoniensis, A. mollis, and A. precatorius. It is one of the characteristic active ingredients of this genus and an important active ingredient of medicinal materials such as Abri Herba. Studies have used abrine as an indicator for the quality control of medicinal materials such as Abri Herba. Modern pharmacological studies have demonstrated that abrine exhibits various pharmacological activities, including anti-tumor, anti-inflammatory, anti-oxidative, and anti-depressive effects. It holds broad prospects for development and application in the prevention and treatment of malignant tumors, inflammatory diseases, liver diseases, renal ischemia-reperfusion injury, and depression-related neuropsychiatric disorders. This article reviewed the research progress in abrine regarding the extraction, detection, and analysis methods, pharmacological effects and mechanisms, and metabolism, aiming to provide reference and data support for new drug development and clinical transformation.
  • Research progress on structure-activity relationship analysis and detection methods of etomidate*
    WANG Shu-ting, WANG Xiao-sen, LÜ Jun, JIN Hong-tao
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1180-1189. https://doi.org/10.16155/j.0254-1793.2025-0679
    Abstract ( )   Knowledge map   Save
    In recent years, the abuse of etomidate has become increasingly serious. With etomidate being listed in the control catalog, the synthesis of non-scheduled analogues by modifying the chemical structure has become an emerging abuse trend to evade regulation, posing a serious threat to public health and safety. In the face of the increasing types of abuse, it is particularly important to strengthen the regulation and detection of etomidate and its analogues. Based on a systematic summary of the pharmacological and toxicological properties of etomidate, this paper focuses on the structure-activity relationships and detection methods of etomidate and its analogues, with the expectation of providing references for the establishment of control strategies and analytical methods for etomidate.
    • Ingredient Analysis
  • Analysis of in vitro dissolution changes of 13 chemical components in Tripterygii Wilfordii Radix and Pteris Multifidae Herba combinations with different ratios based on UPLC-TQ/MS*
    ZHOU Guo-liang, YU Hao, CHEN Hao, WANG Yu, SHANG Er-xin, SU Shu-lan, DUAN Jin-ao
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1190-1201. https://doi.org/10.16155/j.0254-1793.2025-0661
    Abstract ( )   Knowledge map   Save
    Objective: To establish an ultra-performance liquid chromatography-triple quadrupole-mass spectrometry (UPLC-TQ/MS) method for the simultaneous determination of 13 chemical components in Tripterygii Wilfordii Radix (TP) and Pteris Multifidae Herba (PM) and to quantitatively analyze the dissolution changes of chemical components in TP-PM combinations with different ratios. From the perspective of chemical components, this study analyzes the dissolution changes of chemical components in TP-PM combinations with different ratios to determine the optimal compatibility ratio, aiming to provide a basis for elucidating the dose-effect relationship and guiding clinical medication. Methods: Chemical components in TP-PM combinations with different ratios (1 ∶ 4, 1 ∶ 2, 1 ∶ 1, 2 ∶ 1, and 4 ∶ 1) were extracted with water and 70% ethanol, respectively. The dissolution rates of 13 chemical components across the different compatibility ratio groups were analyzed by the UPLC-TQ/MS method. An AcquityTM UPLC BEH C18 chromatographic column (2.1 mm×50 mm, 1.7 μm) was used for gradient elution with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution at a flow rate of 0.3 mL · min-1. Detection was performed through electrospray ionization (ESI+) in positive and negative ion modes (ESI-) with multiple reaction monitoring (MRM). Results: After extraction with 70% ethanol, the total dissolution rates of six terpenoids in the TP-PM combinations with different ratios were all lower than that in TP alone 70% ethanol extract (group C0) (P<0.05). The total dissolution rates of six terpenoids in the water extracts of TP-PM combinations were lower than those in the 70% ethanol extracts. In particular, the dissolution rate of six terpenoids in the TP-PM (1 ∶ 2) water extract group (group S2) was the lowest. Whether water extraction or 70% ethanol extraction, the dissolution rates of all TP-PM combination groups were higher than those of the TP water extract (group S0) and group C0 (P<0.01). Compatibility promoted the dissolution of wilforine, and the dissolution rate was the highest in the group S2. The total dissolution rates of six flavonoids in the TP-PM combinations extracted with 70% ethanol were lower than that of the 70% ethanol extract of PM (group FC0) (P<0.05). The total dissolution rates of six flavonoids in the TP-PM combinations extracted with water were higher than those extracted with 70% ethanol and higher than that in the PM water extract (group FS0) (P<0.01). The total dissolution rate of six flavonoids was the highest in the group S2. Conclusion: Both water extraction and 70% ethanol extraction significantly affect the dissolution rates of six terpenoids, wilforine, and six flavonoids in the TP-PM combinations with different ratios, with TP-PM (1 ∶ 2) water extraction being the most effective. The findings provide a scientific basis for the safe clinical use of the drugs.
  • Quality evaluation of compound Shihu Xiaoke granules based on fingerprinting, quantitative analysis of 21 components, chemometrics, and entropy weight-TOPSIS method*
    ZHENG Wen-wei, WANG Li-na, XIE Jing-xin, ZHOU Ji-xuan, ZHANG Kuan, YAN Jin-xian
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1202-1215. https://doi.org/10.16155/j.0254-1793.2025-0545
    Abstract ( )   Knowledge map   Save
    Objective: To establish a quality evaluation method for compound Shihu Xiaoke granules by combining HPLC fingerprinting, quantitative analysis of 21 components, chemometrics, and entropy-weighted TOPSIS, thereby providing a basis for improving its quality standards. Methods: A Thermo Scientific AccucoreTM XL C18 column (250 mm×4.6 mm, 4 μm) was used at 30 ℃ for gradient elution with a mobile phase of acetonitrile-0.2% phosphoric acid aqueous solution at a flow rate of 1.0 mL · min-1, a detection wavelength of 265 nm, and an injection volume of 10 μL. The HPLC fingerprints of compound Shihu Xiaoke granules were established, and a quantitative analysis method for 21 components was developed and validated. With the obtained data, hierarchical cluster analysis (HCA), principal component analysis (PCA), orthogonal partial least squares-discrimination analysis (OPLS-DA), and entropy-weighted TOPSIS were then employed to evaluate the quality of 15 batches of samples. Results: Methodological validation confirmed that the established fingerprints and quantitative analysis methods met analytical requirements. The similarity of fingerprints for 15 batches of compound Shihu Xiaoke granules ranged from 0.982 to 0.999, with 36 common peaks, of which 21 were identified. HCA, PCA, and OPLS-DA yielded consistent results, clustering the 15 batches into three main categories. With the threshold of variable importance for the projection (VIP)>1, 22 potential differential markers were screened out. The average content of the 21 quantified components—gallic acid, danshensu, protocatechuic acid, protocatechualdehyde, 3'-hydroxypuerarin, caffeic acid, puerarin, mangiferin, 3'-methoxypuerarin, puerarin-6"-O-glucoside, puerarin apioside, daidzin, glycitin, calycosin glucoside, genistin, rosmarinic acid, lithospermic acid, daidzein, salvianolic acid B, calycosin, and salvianolic acid A—ranged from 0.012 mg · g-1 to 2.129 mg · g-1. The entropy-weighted TOPSIS evaluation indicated that batch S5 had the best quality. Conclusion: The established fingerprint and quantitative analysis method for 21 components are accurate, reliable, and reproducible. The approach combining HPLC fingerprinting, quantitative analysis of 21 components, chemometrics, and entropy-weighted TOPSIS enables comprehensive and objective quality evaluation of compound Shihu Xiaoke granules, providing a new technical means for quality control of this preparation.
  • Simultaneous determination of 13 components in Mingmu Jili pills by UHPLC-MS/MS*
    WANG Xiao-fang, ZHAI Si-qi, ZHENG Qian-qian, ZHANG Li-ping, LIU Lin
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1216-1224. https://doi.org/10.16155/j.0254-1793.2025-0531
    Abstract ( )   Knowledge map   Save
    Objective: To establish a ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of 13 chemical components (terrestrosin K, phellodendrine hydrochloride, imperatorin, isoimperatorin, prim-O-glucosylcimifugin, buddleoside, verbascoside, liquiritin, wogonin, geniposide, luteolin-7-O-glucoside, paeoniflorin, and baicalein) in Mingmu Jili pills. Methods: The separation was performed through a Waters Acquity BEH C18 column (100 mm×2.1 mm, 1.7 µm) with a column temperature at 40 ℃. The mobile phase consisted of acetonitrile-5 mmol · L-1 ammonium formate (containing 0.1% formic acid) with gradient elution at a flow rate of 0.3 mL · min-1. The injection volume was 1 µL. The detection was carried out by electrospray ionization (ESI) source combined with multiple reaction monitoring (MRM) in positive and negative ion modes. Results: The 13 components showed good linear relationships in their respective concentration ranges (r>0.999 0). The method demonstrated high precision, repeatability, and stability. The spiked recovery rates ranged from 93.7% to 110.0%, with the relative standard deviation (RSD) of 0.28%-9.7%. The content of 13 components in 13 batches of Mingmu Jili pills was 11.94-75.55, 66.88-156.32, 57.18-87.83, 18.57-27.68, 20.20-196.20, 230.73-485.25, 539.71-3 061.53, 24.30-87.02, 314.32-973.51, 297.45-1 736.38, 61.45-473.48, 397.44-1 275.18, and 331.81-4 091.77 µg · g-1. Conclusion: The established method is efficient, simple, accurate, and reproducible, which can provide a reference for the quality control of Mingmu Jili pills.
    • Metabolism Analysis & Activity Analysis
  • Analysis of constituents in Zaoshi Huatan Xiefei formula in vivo and in vitro based on UPLC-Q Exactive Orbitrap MS/MS*
    CAO Hai-yang, SHAN Bai-xi, ZHAO Peng, DU Yan, LIU Xin-guang, ZHANG Qin, JIANG Yu-hang, ZHANG Zhi-jing, LI Jian-sheng
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1225-1245. https://doi.org/10.16155/j.0254-1793.2025-0405
    Abstract ( )   Knowledge map   Save
    Objective: To qualitatively analyze the chemical components of Zaoshi Huatan Xiefei formula, as well as the components absorbed into the blood and lung based on ultra-high performance liquid chromatography coupled with Quadrupole/Electrostatic field Orbitrap high-resolution mass spectrometry (UPLC-Q Exactive Orbitrap MS/MS). Methods: Chromatographic separation was performed on a Phenomenex Synergi™ Polar-RP column (2 mm×150 mm, 4 μm) with a mobile phase consisting of 0.1% aqueous formic acid (A) and 0.1% formic acid in acetonitrile (B). Gradient elution (0-1 min, 0%B; 1-40 min, 0%B→85%B; 40-43 min, 85%B→100%B; 43-44 min, 100%B→0%B; 44-45 min, 0%B) was conducted with the flow rate of 0.3 mL · min-1, the column temperature maintained at 40 ℃, and the injection volume of 5 µL. An electrospray ionization (ESI) source in both positive and negative ion full-scan mode was used for data acquisition, with a scanning range of m/z 114-1 700 and collision energy of 20, 40, and 60 eV. Chemical components of Zaoshi Huatan Xiefei formula and the components absorbed into the blood and lung were identified by comparing retention time, accurate relative molecular mass, MS/MS fragment ions, reference standards, and relevant literature data. Results: A total of 192 compounds were identified or tentatively characterized in Zaoshi Huatan Xiefei formula, including 41 flavonoids, 45 organic acids, 9 alkaloids, 13 amino acids, 19 phenylpropanoids, 17 terpenoids, 11 saponins, and 37 compounds belonging to other categories. Seventeen compounds were confirmed by comparison with reference standards. Sixty-six prototype components were detected in plasma, and twenty-four were detected in lung tissue. Additionally, the fragmentation pathways of naringin, chlorogenic acid, trigonelline, cryptotanshinone, and magnolol were analyzed. Conclusion: This study comprehensively and rapidly characterizes the diverse chemical components of Zaoshi Huatan Xiefei formula and identifies the components absorbed into the blood and lung. The findings provide a basis for understanding the pharmacological material basis of Zaoshi Huatan Xiefei formula, supporting its clinical application and facilitating new drug development.
  • Component analysis of Ganlu Xiaodu Dan in vivo and in vitro based on UHPLC-Q Exactive Orbitrap HRMS*
    LIU Bin, FENG Xue, ZHANG Cai-juan, WANG Dun-fang, LIU Hai-fan, ZHU Lin, LIU Jian-yao, SUN Qi-yue, YANG Wei-peng
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1246-1257. https://doi.org/10.16155/j.0254-1793.2025-0421
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    Objective: To conduct a qualitative study on the chemical components of Ganlu Xiaodu Dan and its blood- and brain-entering components using ultra-high-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry (UHPLC-Q Exactive Orbitrap HRMS). Methods: UHPLC-Q Exactive Orbitrap HRMS was employed to identify the chemical components of Ganlu Xiaodu Dan. Twelve male Sprague-Dawley (SD) rats were randomly divided into a blank group and a Ganlu Xiaodu Dan group. The Ganlu Xiaodu Dan group received an intragastric administration of the aqueous decoction at a raw herb dose of 1.8 g · kg-¹, while the blank group received an equivalent volume of purified water. Plasma samples were collected 1.5 h post-administration. Using an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 µm) at a column temperature of 40 ℃. The mobile phase consisted of 0.1% formic acid in water and acetonitrile with gradient elution at flow rate of 0.30 mL · min-1. The injection volume was 6 µL. Electrospray ionization (ESI) was applied with a scanning range of m/z 100-1 500. Combined with information from a reference database, qualitative analysis of blood-entering components was performed in both positive and negative ion modes. Results: A total of 195 compounds were identified in the 80% methanol extract of Ganlu Xiaodu Dan, and 66 prototype components were detected in serum. The fragmentation patterns of baicalin, apigenin-7-glucoside, isorhamnetin-3-O-galactoside, biochanin A, 28-deacetylbelamcandal, plantamajoside A, phillyrin A, aurantio-obtusin, neochlorogenic acid, chrysosplenol, mangiferin, and chuanbeiketonine were analyzed. Conclusion: This study enabled the rapid characterization and identification of multi-category in vitro chemical components and blood-entering components of Ganlu Xiaodu Dan, providing a reference for research on its pharmacodynamic material basis and quality control.
  • HPLC fingerprints and glucose-lowering activity-fingerprint relationship of Lycii Cortex
    LIU Yu-ting, HOU Jia-hao, WANG Chao-hui, YU Yin-feng, YANG Ji-xia, SONG Yong-xing, FENG Chao, MA Dong-lai
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1258-1271. https://doi.org/10.16155/j.0254-1793.2025-0498
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    Objective: To establish the HPLC fingerprints of Lycii Cortex from different origins, study the glucose-lowering activity-fingerprint relationship of Lycii Cortex, and preliminarily clarify the main active ingredients of Lycii Cortex for lowering glucose. Method: A Zorbax SB-AQ C18 column (250 mm×4.6 mm, 5 μm) at a column temperature of 40 ℃ was used for gradient elution with a mobile phase consisting of acetonitrile-0.1% trifluoroacetic acid aqueous solution at a flow rate of 1.0 mL · min-1. The detection wavelength was set as 280 nm, and the injection volume was 10 μL. HPLC fingerprinting was performed for nine components—caffeoylputrescine, chlorogenic acid, scopolin, vanillic acid, kukoamine B, kukoamine A, ferulic acid, cinnamic acid, and N-trans-feruloyltyramine—in 25 batches of Lycii Cortex samples from different geographical origins. Hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were employed to distinguish and compare Lycii Cortex samples from various origins and to identify the main differential components. An insulin-resistant HepG2 (IR-HepG2) cell model was established with insulin, and the glucose oxidase method and CCK-8 assay were used to determine the appropriate treatment concentration of Lycii Cortex for this model. Glycogen content, hexokinase (HK) activity, and pyruvate kinase (PK) activity were measured after treatment with Lycii Cortex samples from different origins. An activity-fingerprint relationship model linking the marker components of Lycii Cortex from various origins to their glucose-lowering effects on IR-HepG2 cells was constructed through grey relational analysis and partial least squares regression (PLSR). Results: The HPLC fingerprints of Lycii Cortex were constructed, showing 28 common peaks, among which 9 chemical components were identified. The results of grey relational analysis indicated that the glucose-lowering effect was a synergistic effect of multiple components. The results of PLSR showed that there were six peaks with VIP>1, including peaks 9 (kukoamine B), 4, 10 (kukoamine A), 12, 5 (caffeoylputrescine), and 1, which were positively correlated with glucose-lowering activity. Conclusion: The analysis of the activity-fingerprint relationship preliminarily clarifies the glucose-lowering substance basis of Lycii Cortex and provides a scientific reference for its quality control.
    • Bioassay
  • A sequence study of CpG oligodeoxynucleotide vaccine adjuvants based on UPLC-Q TOF MS/MS*
    LI Dan-xi, WANG Jin, LIU Bo, WANG Yi-ping, NING Bao-ming, YANG Hui-ying, WU Xian-fu, SUN Hui-min
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1272-1281. https://doi.org/10.16155/j.0254-1793.2026-0072
    Abstract ( )   Knowledge map   Save
    Objective: To establish an analytical method based on ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q TOF MS/MS) for the sequence determination of CpG oligodeoxynucleotide (CpG ODN) immune adjuvants, thereby providing technical support for their quality control and quality consistency evaluation. Methods: The CpG ODN HP 007 was used as the sample for analysis by UPLC coupled with high-resolution mass spectrometry. The trimethylamine-hexafluoroisopropanol ion pair system was utilized as the mobile phase, which was separated by gradient elution through the C18 column. The MS data were collected in the electrospray negative ion mode to obtain accurate molecular weight information. On the basis of screening of different charge state parent ions, a single specific parent ion independent targeted acquisition strategy was employed to obtain characteristic fragments from the MS2 spectrum. Subsequent to data acquisition, deconvolution, fragment ion recognition, and sequence matching were carried out. Thereafter, a multi-dimensional index comprehensive judgment framework integrating sequence coverage, fragment matching continuity, and confirmation status was constructed to confirm the target sequence. Results: The measured molecular mass of HP 007 was 9 345.51, which closely matched the theoretical molecular mass of the target object sequence (9 345.5). The fragment information generated by different charge state parent ions exhibited significant complementarity. The implementation of a multi-parameter comprehensive judgment strategy has been demonstrated to facilitate the reliable confirmation of target sequence. Conclusion: The UPLC-Q TOF MS/MS sequence confirmation method integrating multi-charge state parent ion analysis and multi-parameter comprehensive evaluation established in this study enhances the reliability of ODN sequence confirmation. It compensates for the limitations of the single coverage index in sequence determination. Furthermore, it can serve as a reference for sequence confirmation and quality control of CpG ODN vaccine adjuvants.
  • Establishment of detection method for anti-drug antibodies to insulin glargine biosimilar*
    LIU Qing, LI Sha-sha, YAO Lin, CHANG Rui-heng, ZHANG Chun-yun, HU Jin-fang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1282-1289. https://doi.org/10.16155/j.0254-1793.2025-0544
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    Objective: This paper aims to establish a single bioanalytical method applicable for detecting anti-drug antibodies to insulin glargine and its biosimilar. Methods: The enzyme-linked immunosorbent assay was established by using biosimilars as both the capture antigen and the competitive antigen in the confirmation assay, as well as peroxidase-conjugated recombinant protein A/G as the detection reagent. The method validation was carried out from the aspects of antigen equivalence, screening cutoff, titer cutoff, confirmatory cutoff, sensitivity, precision, selectivity, hook effect, stability, and system adaptability. Results: Antigen equivalence (titration similarity and inhibition similarity) cross validation showed that the binding ability of biosimilars and reference drugs exhibited similar binding ability to positive antibodies. The screening cutoff factor was 1.186; the titer cutoff factor was 1.454; the confirmatory cutoff was 10.7%. The sensitivities of the screening assay and confirmatory assay were 8.9 ng · mL-1 and 7.7 ng · mL-1, respectively. The intra-plate and inter-plate relative standard deviations (RSDs) were less than 20% (The RSD of intra-plate precision of inhibition rate in the negative samples in the confirmation assay was 20.0%-22.8%, excluded). There was no hook effect in the range of 0.8-6.4 μg · mL-1. The samples were stable at room temperature for 3 h, refrigerated (2-8 ℃) for 1 d, and at -70 ℃ after four freeze-thaw cycles. Selectivity, drug tolerance, and system suitability were satisfactory. Conclusion: In this study, a single bioanalytical method applicable for detecting anti-drug antibodies to insulin glargine and its biosimilar is successfully established by the antigen equivalence verification of biosimilar and reference drugs.
  • Microbial monitoring analysis of pharmaceutical water systems and preliminary exploration of subculture in three enterprises*
    HE Zhi-yong, ZHANG Xu-hong, FANG Li-na, ZHENG Xiao-ling
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1290-1298. https://doi.org/10.16155/j.0254-1793.2025-0723
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    Objective: To conduct microbial monitoring and analysis of pharmaceutical water systems from three enterprises, and preliminarily explore culture media suitable for subculture of microorganisms isolated from these systems. Methods: A total of 130 microbial strains were isolated from the pharmaceutical water systems of three enterprises. Bacterial and fungal isolates were identified by 16S rDNA and ITS rDNA sequencing, respectively. Phylogenetic trees were constructed for frequently detected genera to analyze potential contamination sources. Furthermore, subculture studies were performed on all the 130 strains with three different culture media and the modified sterilization method. Results: The microorganisms most frequently detected in the water systems of the three pharmaceutical enterprises were Methylobacterium, Ralstonia, and Bradyrhizobium. Subculture results of the 130 strains showed that R3A and R2A media performed better than Tryptic Soy Agar (TSA). The sterilization method was modified as autoclaving phosphate and agar components separately before mixing in the R3A medium, which improved the growth rate of some slow growing microorganisms on RXA media. Conclusion: Microorganisms in pharmaceutical water systems show certain commonality. RXA media may be more suitable for subculture of microorganisms from pharmaceutical water systems. For some microorganisms susceptible to oxidative stress, modifying the sterilization method for the culture medium can promote their growth.
  • Determination of free fatty acid content in antibody pharmaceuticals using LC-MS technology
    OUYANG Zi-jun, XING Zhan-lei, HU Nan, HUANG De-feng, WANG Hui, CHEN Xi, LI Gang, SONG Lan-kun
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1299-1306. https://doi.org/10.16155/j.0254-1793.2025-0500
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    Objective: To develop a LC-MS method for the quantitation of three free fatty acids—lauric acid, myristic acid, and oleic acid—in antibody pharmaceuticals based on the analytical quality by design (AQbD) principles outlined in the ICH Q14 guideline on analytical procedure development, and to perform subsequent method validation. Methods: The analytical target profile (ATP) was initially established. Based on literature review and laboratory experience, a 1290 UHPLC (ultra-high performance liquid chromatography) system coupled with an SQ- MS (single quadrupole mass spectrometer) was selected to develop the method. All operational steps, spanning from sample preparation to LC-MS detection, that could potentially influence analytical method performance were comprehensively identified, and their potential impact on method accuracy and precision was systematically assessed. With respect to the control strategy, factors associated with high risk or high operational cost were subjected to stringent control. Experimental investigations were conducted on chromatographic column selection, mobile phase composition, gradient or isocratic elution modes, column temperature, as well as mass spectrometric parameters including capillary voltage, fragmentor voltage, and nebulizer pressure. A quantitative analytical method suitable for the degradation products of polysorbate (Tween) was developed and subsequently validated in accordance with ICH Q2 requirements. Results: Methodvalidation demonstrated that the established LC-MS method for free fatty acid quantification exhibited spike recoveries ranging from 82.3% to 106.6% for lauric acid, myristic acid, and oleic acid. The linear range spanned from 0.01 to 2.00 μg · mL-1, with a lower limit of quantitation (LLOQ) below 0.01 μg · mL-1. No interference from proteinaceous components or excipients within the antibody formulation was observed. The method was applied to analyze the concentrations of lauric acid, myristic acid and oleic acid in both within-specification and expired antibody preparations, as well as in forced degradation samples from two investigational antibody formulations. Notably, significantly elevated levels of lauric acid, myristic acid, and oleic acid were detected in the forced degradation samples, indicative of polysorbate degradation. The method performance met the predefined requirements specified in the ATP, rendering it suitable for the quantitative detection of free fatty acids in diverse antibody pharmaceuticals. Conclusion: This study presents a case study of LC-MS method development employing AQbD principles. Through rigorous control of experimental materials, measurement procedures, and operational protocols, and via experimental verification of permissible ranges for mass spectrometer gas flow and voltage parameters, the final method specifies mass spectrometric parameters as acceptable operational ranges rather than discrete fixed values. This approach holds substantial value for the method lifecycle, particularly in the context of change management. The developed method is applicable to the quantification of free fatty acids derived from polysorbate degradation in antibody products. Furthermore, this case provides a meaningful reference for the application of LC-MS methodology in pharmaceutical quality control.
  • Optimization and validation of the specific neutralization test method for Corynebacterium diphtheriae strain C7 (β197)*
    DONG Guo-xia, SHI Bo-zhi, TAN Ya-jun, XUE Yan-hua, TIAN Lin, SHI Ji-chun, YUAN Li-yong
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1307-1311. https://doi.org/10.16155/j.0254-1793.2025-0698
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    Objective: To optimize and validate the specific neutralization test method for the Corynebacterium diphtheriae strain C7 (β197) (strain C7), thereby improving the clarity of results and shortening the detection time. Methods: Strain C7 was inoculated in both wide and narrow patterns. A filter paper strip was placed either on the surface of culture medium or embedded within the culture medium. Seven types of culture media—peptone A Elek’s medium, peptone B Elek’s medium, peptone C Elek’s medium, peptone D Elek’s medium, peptone E Elek’s medium, nutrient agar medium, and commercially available Elek’s medium—were screened. Diphtheria antitoxin was prepared at four concentrations: 2 000, 1 500, 1 000, and 500 Lf · mL-1. Petri dishes were poured with 15, 25, and 35 mL of culture medium, and strain C7 was inoculated at distances of 3, 5, 7, 9, and 11 mm from the filter paper strip. Equine serum or bovine serum was added to the basal culture medium. The key conditions for the specific neutralization test of strain C7 were systematically optimized and screened. Additionally, the specificity and repeatability of the optimized method were validated. Results: The optimal conditions for the specific neutralization test were determined as follows: strain C7 inoculated in a narrow pattern, filter paper strip placed on the surface of culture medium, peptone C Elek’s medium used as the neutralization medium, filter paper strip saturated with diphtheria antitoxin at 1 500 Lf · mL-1, petri dish poured with 15 mL medium, strain C7 inoculated at a distance of 3 mm from the filter paper strip, and bovine serum added to the basal medium. The validation results demonstrated that the optimized method had good specificity and repeatability. Conclusion: The specific neutralization test method optimized in this study is stable and reliable. It significantly improves the clarity of results and detection efficiency for strain C7, providing more efficient and reliable technical support for the laboratory identification of this bacterium and the quality control of related vaccines.
    • Safety Monitoring
  • Analysis of heavy metals, arsenic speciation, and valence states in Rukuaixiao preparations based on HPLC-ICP-MS*
    ZHAO Qi, CHEN Rong, WANG Ya-qiong, ZHANG Ya-shuang, ZHU Meng
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1312-1319. https://doi.org/10.16155/j.0254-1793.2025-0371
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    Objective: To study the heavy metals and harmful elements and the arsenic (As) speciation in Rukuaixiao preparations and evaluate the As risk. Methods: Residues of Cr, Ni, Cu, As, Cd, Hg, and Pb in 78 batches of Rukuaixiao preparations were determined by microwave digestion-inductively coupled plasma-mass spectrometry (ICP-MS). High performance liquid chromatography coupled with inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) was employed to analyze six As forms. The hazard index (HI) was adopted to evaluate the risk of As in 10 batches of samples. Results: The two methods demonstrated good linear relationship, precision, repeatability, and recovery. The content of seven heavy metals varied greatly among the samples from different manufacturers. The As speciation and valence states in 10 batches of samples from E company were analyzed. The results showed that total As content was high. HPLC-ICP-MS results showed that trivalent and pentavalent As exhibited high toxicity. The HI<1 indicated that the As risk was controllable. Conclusion: The established method for the determination of seven heavy metals and harmful elements and six forms of As demonstrates high efficiency, rapidity, accuracy, and stability. This study provides data support for the formulation of the limit standards for the heavy metals and harmful elements in Rukuaixiao preparations.
  • An examination of the electrochemical properties of As (Ⅲ) in Realgar via electrochemical techniques*
    LIU Jing-yu, DING Yi, LI Ya-nan, ZHANG Dan-dan, ZHANG Yun-zheng, ZHANG Yu-bi, WANG Xian-shu
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1320-1329. https://doi.org/10.16155/j.0254-1793.2025-0595
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    Objective: To establish a rapid and sensitive method for determining As (Ⅲ) in Realgar, a commonly used Chinese medicinal material with safety mainly influenced by As (Ⅲ), thereby ensuring its safe use. Methods: An iron-cobalt layered double hydroxide/transition metal carbide (Fe-Co-LDH/MXene) nanocomposite was synthesized by a hydrothermal method. A glassy carbon electrode (GCE) was modified with this nanocomposite to fabricate a sensor (Fe-Co-LDH-MXene/GCE). The materials were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). The sensor performance under different solution conditions was systematically evaluated by electrochemical methods. Furthermore, the stability and repeatability were assessed. Results: A nanocomposite with a distinct layered structure was successfully prepared. In the neutral aqueous solution, the sensor demonstrated excellent electrochemical responses to As (Ⅲ). A good linear relationship was observed between the response signal and As (Ⅲ) concentration within the range of 100-500 μg · mL-1. The sensor showed good stability and repeatability. Moreover, it performed well in the determination of As (Ⅲ) in Realgar samples. Conclusion: The electrochemical sensor enables rapid and accurate determination of As (Ⅲ) in Realgar under neutral conditions, with simple operation and high sensitivity. It offers a new method for quality control of Realgar-containing Chinese medicine preparations.
  • A gene membrane chip method for rapid screening of six pathogenic bacteria in cosmetics*
    YANG Hong-xia, LIAO Hui, ZHANG Lu, WANG Zhi-jian, LI Jue, ZHENG Xiao-ling, ZHANG Lin-shuang, CAO Jie
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1330-1336. https://doi.org/10.16155/j.0254-1793.2025-0623
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    Objective: To establish a rapid coculture and detection method for six pathogenic bacteria (Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella paratyphi B, Pseudomonas aeruginosa, and Burkholderia cepacia) in cosmetics and raw materials based on the gene membrane chip technology and improve the biological safety assurance capability and risk screening efficiency of the product. Methods: The co-culture medium was used for co-culture of the six pathogenic bacteria, and specific primers and probes for the six pathogenic bacteria were designed. Multiplex PCR amplification was performed on the sample nucleic acid, and the gene membrane chip technology was employed for further product hybridization and analysis. Results: Specificity tests showed that the six pathogenic bacteria developed color only at their corresponding target spots, while the four negative template bacteria (Listeria monocytogenes, Bacillus cereus, Vibrio vulnificus, and Shigella spp.) showed no color development, indicating no cross-reaction and good probe specificity. At primer concentrations of 0.20/0.30 μmol · L-1 (internal control) and 0.067-0.75 μmol · L-1 (for the six targets), the color development intensities of all targets were uniform. The limit of detection (LOD) test showed that the established method achieved an LOD of 1.0×105 copies · mL-1 for all six pathogenic bacteria. The test results of nine batches of samples were all negative, consistent with those obtained by conventional methods. The established method specifically detected the corresponding targets in the spiked samples without cross-interference, meeting the requirements for routine testing. Conclusion: A co-culture method for six pathogenic bacteria in cosmetics and a multiplex detection method based on PCR-gene membrane chip technology were established. This method exhibits strong specificity, good stability, and high sensitivity, ensuring the quality and safety of cosmetics and raw materials. It can be further applied to the detection of cosmetics in factories and markets, enriching the detection methods for pathogenic bacteria in the cosmetics industry and providing a rapid and powerful technical means for detection.
  • Influencing factors in determination of aminoglycoside antibiotics in water by UPLC-QQQ MS/MS
    LIU Bin, HUANG Wan, GUO Li, WANG Fan, WU Bing-chan
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1337-1345. https://doi.org/10.16155/j.0254-1793.2025-0625
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    Objective: To establish an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ MS/MS) method for the simultaneous determination of 10 aminoglycoside antibiotics in water. Methods: Following alkaline desorption of particulates in the water sample, heptafluorobutyric acid was added to form an ion pair with the target analytes, and then the ion pair was determined by the UPLC-QQQ MS/MS method. The analytes were separated on a Poroshell 120 SB-C8 chromatographic column (100 mm×3.0 mm, 2.7 μm) by gradient elution, with the mobile phase of 1 mmol · L-1 ammonium acetate and 0.1% (volume fraction) formic acid aqueous solution and acetonitrile. Then, the target analytes were ionized by electrospray ionization in positive ion mode, detected in multiple reaction monitoring mode, and quantified by the internal standard method. Key influencing factors, including sample stability, storage container, particulate matter in samples, solution pH, filter membranes, residual chlorine content in samples, and liquid chromatographic conditions, were systematically evaluated, and spiked water samples of five types of matrices were determined. Results: Samples can be stored sealed, refrigerated, and protected from light at below 4 ℃ for at least seven days, and plastic containers shall be used for storage. Aminoglycoside antibiotics tended to be adsorbed onto particulate matter, so alkaline desorption was required, and the filtrate must be adjusted to neutral before determination. A 0.22 μm polypropylene filter membrane shall be used for sample filtration. Residual chlorine can cause loss of the target analytes, and ascorbic acid must be added immediately after water sample collection. After optimizing instrumental conditions, limits of detection ranged from 1.0 to 1.7 μg · L-1. Recovery tests were conducted using the standard addition method. Except for spectinomycin, which showed slightly lower recoveries in complex matrices (with average recoveries of 42.8% and 41.0% in domestic sewage and industrial wastewater, respectively), the recoveries of the other compounds in all matrices, as well as those of spectinomycin in pure water, groundwater, and surface water matrices, ranged from 60.0% to 127.0%. The relative standard deviations (RSDs) (n=6) were between 1.3% and 16.0%. Conclusion: All performance parameters of this method satisfy current quality-control requirements for environmental antibiotic monitoring and thus are of practical referential significance.
  • Determination of 31 chemical drugs in disinfection products by HPLC-MS/MS*
    ZHANG Yan-chao, ZHANG Meng, LIU Jing, LIU Hong-xiang
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1346-1353. https://doi.org/10.16155/j.0254-1793.2025-0672
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    Objective: To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous determination of 31 chemical drugs in disinfection products. Methods: A Waters ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) with the column temperature of 30 ℃ was used for gradient elution with the mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in acetonitrile at a flow rate of 0.3 mL · min-1. The injection volume was 2 μL. Mass spectrometry acquisition was conducted in positive and negative ion modes with electrospray ionization (ESI) and analyses were detected in the multiple reaction monitoring (MRM) mode. Results: The 31 chemical drugs were effectively separated within 13 min, and the limit of detection was 0.05 mg · kg-1. A good linear relationship (r>0.99) was observed between peak area and concentration within the range of 0.125-2.5 mg · kg-1. The spiked recovery rate ranged from 73.2% to 114.0%, with the relative standard deviation (RSD)≤12.2% (n=6). Conclusion: This method is rapid, accurate, sensitive, and specific, allowing the simultaneous detection of 31 chemical drugs in disinfection products and providing technical reference for the regulation of disinfection products.
    • Standard Deliberation
  • Establishment of the specific chromatogram of Qizhi Tongluo capsules based on UPLC-QQQ MS
    LI Ting, LI Min, LEI Zhen-hong, LI An-ping, CHENG Xian-long
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1354-1362. https://doi.org/10.16155/j.0254-1793.2025-0590
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    Objective: To establish an ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ MS) method for the rapid and simple qualitative identification of Qizhi Tongluo capsules. Methods: Gradient elution was performed through a Waters ACQUITYTM UPLC Hss T3 C18 column (100 mm×2.1 mm, 1.8 μm) with the mobile phase composed of acetonitrile and 0.1% formic acid aqueous solution at the flow rate of 0.3 mL · min-1 and the column temperature of 35 ℃. The chromatographic effluents were detected by mass spectrometry in positive and negative ion modes. Results: The specific chromatograms showed 32 common characteristic peaks from 13 Chinese herbal medicines. The structures of 23 characteristic components were confirmed by comparison with reference substances. RSDs of the relative retention time of 32 characteristic peaks in 10 batches of samples were all less than 1.0%, indicating that the 32 characteristic peaks were stable. Conclusion: The established liquid chromatography-mass spectrometry (LC-MS) specific chromatogram method is simple, sensitive, and accurate, comprehensively reflecting the chemical composition characteristics of Qizhi Tongluo capsules and serving as a new method for rapid identification of Qizhi Tongluo capsules.
  • Development of a RT-qPCR assay for detection of Burkholderia cepacia complex based on the hutC gene*
    YU Meng, WANG Si-jin, ZHANG Xun-di, MA Shi-hong
    Chinese Journal of Pharmaceutical Analysis. 2026, 46(7): 1363-1372. https://doi.org/10.16155/j.0254-1793.2025-0445
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    Objective: To rapidly identify the contamination risk from Burkholderia cepacia complex (Bcc) during pharmaceutical production, a real-time fluorescent quantitative PCR (RT-qPCR) detection method based on the hutC gene was developed. Methods: The nucleotide sequences of the hutC gene from Bcc and its related species were analyzed. Specific TaqMan probes and primers targeting the conserved regions of the hutC gene were designed, and an internal amplification control (IAC) probe was also constructed. The reaction system and conditions were optimized. The established method was validated using Bcc and non-Bcc strains, and parameters including specificity, amplification efficiency, and detection limit were evaluated. Both pharmacopoeia methods and the established RT-qPCR methods were used to detect Bcc in 20 artificially contaminated water-based products. Results: In the optimized reaction system, the final concentrations of both forward and reverse primers were 200 nmol · L-1; the final concentrations of hutC probe and IAC probe were 200 nmol · L-1 and 400 nmol · L-1, respectively, with 1 μL of IAC template DNA at a concentration of 1.3×10-5 ng · μL-1 added to each reaction. In the experiments, specific amplification was observed for all 12 Bcc strains, while no specific amplification was detected for the 21 non-Bcc strains, including closely related species within the same genus, pharmacopoeia specified control bacteria, and common Gram-negative bacteria isolated from water systems. When IAC was introduced, the amplification efficiency was 101% for the hutC recombinant plasmid and 88% for B. cepacia ATCC 25416 culture. The detection limit was 14 CFU · reaction-1. The qualitative detection results of the 20 artificially contaminated water-based products obtained by the developed method were consistent with those of the pharmacopoeia method, whereas the detection time was reduced from 6-7 d (pharmacopoeia method) to 24 h. Conclusion: The established RT-qPCR method can serve as a risk control approach in pharmaceutical production, enabling rapid and specific detection of Bcc contamination in high-risk water-based products, in-process materials, and raw materials.
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