中国药物警戒 ›› 2025, Vol. 22 ›› Issue (2): 197-205.
DOI: 10.19803/j.1672-8629.20240854

• 安全与合理用药 • 上一篇    下一篇

基于“性状-化学-生物”序贯检测的柴胡饮片质量评控

郑长辉1, 马丽娜2,*, 何婷2, 顾媛媛2, 赵薇2, 汪红斌2, 牛源菲1, 富尧3, 陈彦君4, 曹俊岭2,5#   

  1. 1北京中医药大学中药学院,北京 100029;
    2北京中医药大学东方医院,北京 100078;
    3北京中医药大学东直门医院,北京 100700;
    4深圳市广祥医药有限公司,广东 深圳 518000;
    5北京中医药大学东直门医院洛阳医院,河南 洛阳 471002
  • 收稿日期:2024-11-04 出版日期:2025-02-17 发布日期:2025-02-17
  • 通讯作者: *马丽娜,女,博士,副主任药师,中药质量与药理。E-mail:malina8512@163.com;#为共同通信作者。
  • 作者简介:郑长辉,男,在读硕士,中药质量与药理。
  • 基金资助:
    国家中医药管理局司便函(国中医药科技中药便函[2022]59号); 国家中医药管理局科技司项目（GZY-KJS-2022-057）; 国家自然科学基金资助项目（82104518）; 中央高校基本科研业务费专项（2023-JYB-KYPT-18）

Quality Evaluation and Control of Bupleuri Radix Decoction Pieces Based on ‘Shape-Chemistry-Biology' Sequential Detection

ZHENG Changhui1, MA Lina2,*, HE Ting2, GU Yuanyuan2, ZHAO Wei2, WANG Hongbin2, NIU Yuanfei1, FU Yao3, CHEN Yanjun4, CAO Junling2,5#   

  1. 1School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China;
    2Dongfang Hospital of Beijing University of Chinese Medicine, Beijing 100078, China;
    3Dongzhimen Hospital of Beijing University of Chinese Medicine, Beijing 100700, China;
    4Shenzhen Guangxiang Pharmaceutical Co., Ltd, Shenzhen Guangdong 518000, China;
    5Luoyang Hospital of Dongzhimen Hospital of Beijing University of Chinese Medicine, Luoyang Henan 471002, China
  • Received:2024-11-04 Online:2025-02-17 Published:2025-02-17

PDF (PC)

52

摘要： 目的 探究基于“性状-化学-生物”三位一体的柴胡饮片质量评价体系,保障柴胡饮片临床用药安全有效。方法 网络采购不同药材市场和直接购买正规饮片公司柴胡饮片样品26批,同时制备6批特殊样品（水煮、醇提）,共32批样品。采用德尔菲（Delphi）法对不同来源柴胡饮片进行传统经验鉴别;高效液相色谱法（HPLC）对5种柴胡皂苷（a、b1、b2、c、d）进行含量测定,分析不同来源柴胡饮片指标性成分含量差异,并进行正交偏最小二乘法判别分析（OPLS-DA）和变量重要性投影值分析（VIP）;采用脂多糖诱导的RAW264.7细胞炎症模型,通过检测其对白介素6(IL-6)、肿瘤因子α(TNF-α)炎症因子表达的影响,考察其抗炎活性;测定不同来源柴胡饮片的细胞毒性,明确安全范围。结果 传统性状鉴别可有效识别指标性成分含量较高的藏柴胡和性状不易分辨的竹叶柴胡,特殊制备样品（水煮、醇提）在性状鉴别中未能有效分辨。含量测定结果显示32批柴胡饮片中,除4批特殊制备样品（TS2、TS4、TS5、TS6）柴胡皂苷a、d含量不符合《中华人民共和国药典》（2020年版）规定的0.3%限度外,其余28批样品含量结果均符合《中华人民共和国药典》（2020年版）0.3%的规定。但全部6批特殊制备样品（TS1~TS6）5种皂苷总含量均低于北柴胡样品的最低含量。5种皂苷总含量在北柴胡和藏柴胡中分别为0.53%~1.09%和3.40%~4.01%,藏柴胡为北柴胡的3~4倍,两者掺杂品种的总含量在2.02%~3.41%。水煮、醇提样品5种柴胡皂苷总含量显著下降为原样本的50%、25%。进一步生物评价发现,不同来源柴胡饮片具有明显的抗炎活性（P<0.05、0.01）,藏柴胡抗炎活性优于北柴胡（P<0.05、0.01）,细胞毒性结果显示藏柴胡细胞毒性明显大于北柴胡（P<0.01）;竹叶柴胡抗炎活性和细胞毒性均与北柴胡接近。结论 采用“传统性状-化学含量测定-生物评价”序贯检测的方法,可更全面准确反映柴胡饮片质量,更有效发现不合格样品,保证柴胡饮片质量的一致性和稳定性,提高临床用药有效性和安全性。

关键词: 柴胡, 性状鉴别, 抗炎活性, 细胞毒性, 化学含量测定, 生物评价, 质量评控

Abstract: Objective To explore the quality evaluation system of Bupleuri Radix decoction pieces based on the ‘Shape-Chemistry-Biology' trinity so as to ensure the safety and effectiveness of clinical medications with Bupleuri Radix decoction pieces. Methods Twenty-six batches of Bupleuri Radix decoction pieces from different medicinal materials markets and from licensed decoction piece companies were purchased online, and 6 batches of special samples (water boiled and alcohol extracted) were prepared at the same time, so there were a total of 32 batches of samples. Bupleuri Radix decoction pieces from different sources were identified empirically using the Delphi method. The contents of 5 types of saikosaponins (a, b1, b2, c, d) were determined with HPLC, and the content differences of index components of Bupleuri Radix decoction pieces from different sources were analyzed. Orthogonal partial least squares discriminant analysis (OPLS-DA) and variable importance in projection (VIP) were carried out. The lipopolysaccharide-induced RAW264.7 cell inflammation model was used to investigate its anti-inflammatory activity by detecting its effect on expressions of interleukin 6 (IL-6) and tumor factor α (TNF-α) inflammatory factors. At the same time, the cytotoxicity of Bupleuri Radix decoction pieces from different sources was determined, and the safety range was clarified to effectively identify unqualified samples. Results The traditional trait identification method could effectively identify the Bupleurum marginatum var. stenophyllum with high contents of index components and the Bupleurum marginatum with indistinguishable traits. However, the special prepared samples (boiled and alcohol extracted) could not be effectively distinguished during trait identification. The results showed that the contents of saikosaponin a and d in the 32 batches of Bupleuri Radix decoction pieces except 4 batches of special prepared samples (TS2, TS4, TS5, TS6) did not meet the 0.3 % limit stipulated by the ‘Pharmacopoeia of the People's Republic of China' (2020 edition). However, the total contents of five saponins in all the six batches of special prepared samples (TS1-TS6) were lower than the lowest content of Bupleurum chinense samples, which were 0.53% to 1.09% and 3.40% to 4.01% in Bupleurum chinense and Bupleurum marginatum var. stenophyllum, respectively. The total content of Bupleurum marginatum var. stenophyllum was 3 to 4 times that of Bupleurum chinense, while that of the two doped varieties ranged from 2.02% to 3.41%. The total contents of 5 kinds of saikosaponins in boiled and alcohol-extracted samples decreased significantly to 50% and 25% of the contents of the original samples. Further biological evaluation showed that Bupleuri Radix decoction pieces from different sources had obvious anti-inflammatory activity (P<0.05,0.01), which was better in Bupleurum marginatum var. stenophyllum than in Bupleurum chinense (P<0.05,0.01). The cytotoxicity of Bupleurum marginatum var. stenophyllum was significantly higher than that of Bupleurum chinense (P<0.01). The anti-inflammatory activity and cytotoxicity of Bupleurum marginatum Wall. were close to those of Bupleurum chinense. Conclusion The sequential detection method of ‘traditional traits-chemical content determination-biological evaluation' can more accurately reflect the quality of Bupleuri Radix decoction pieces, more effectively detect unqualified samples, ensure the consistency and stability of the quality of Bupleuri Radix decoction pieces, and improve the effectiveness and safety of clinical medications.

Key words: Bupleuri Radix, Character Identification, Anti-Inflammatory Activity, Cytotoxicity, Chemical Content Determination, Biological Evaluation, Quality Evaluation and Control

中图分类号: