目的 以抗炎为例,建立基于效应成分指数(effect-constituents index,ECI)的金银花配方颗粒质量一致性评价方法。方法 采用超高效液相色谱法测定22批次不同标准(国标/企标)的金银花配方颗粒样品中9种具有抗炎活性的单体成分(新绿原酸、绿原酸、隐绿原酸、断马钱子酸、断氧化马钱子苷、木犀草苷、异绿原酸B、异绿原酸A、异绿原酸C)含量,采用环氧合酶-2(cyclooxygenase-2,COX-2)试剂盒测定上述样品的抗炎效价和相关单体成分的半数抑制浓度(half inhibitory concentration,IC₅₀),进而整合分析获取上述各个成分的抗炎效价权重系数,通过权重系数和效价整合计算得到ECI,最后通过细胞实验验证所构建的ECI的可行性。结果 抗炎成分含量测定结果显示,9种单体成分含量存在较大的差异,其中国标批次(S1~S11)和企标批次(S12~S22)在单体成分的含量高低也存在显著差异,如国标批次(S8)中绿原酸的含量是企标批次(S16)中该成分含量的3.3倍(43.06 vs 12.98 mg/g)。国标(S1~S11)样品的整体稳定性与企标(S12~S22)样品相差不大(RSD_国标=211.55,RSD_企标=215.71)。生物效价测定方法通过了方法学考察,结果显示国标样品的抗炎效价和稳定性比企标更好(效价_国标=100.28~106.35 U/mL,效价_企标=88.06~95.49 U/mL;RSD_国标=1.98,RSD_企标=2.56)。ECI理论计算结果显示该方法结果与实验结果一致,能够反映金银花配方颗粒样品的药效关系(ECI_国标=10.82~14.11,ECI_企标=9.32~12.58;RSD_国标=9.51,RSD_企标=10.02)。相关性分析显示单一指标性成分的含量与抗炎效价之间正相关性较好的是绿原酸(r=0.644),其次是木犀草苷(r=0.581),说明这2个成分在抗炎活性中起主要作用。结论 以抗炎为例,建立基于效应成分指数的金银花配方颗粒质量评价方法。可应用于多批次不同标准(国标/企标)的金银花配方颗粒的质量一致性评价,为其他具有类似抗炎作用的中药配方颗粒的质量评价研究及应用提供科学参考。

Objective To establish a method for quality consistency evaluation of Jinyinhua (Lonicerae Japonicae Flos) dispensing granules based on effect-constituents index (ECI) by taking anti-inflammation as an example. Methods Ultra performance liquid chromatography (UPLC) was used to determine the contents of nine monomer components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, secologanic acid, secoxyloganin, cymaroside, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C) with anti-inflammatory activity in 22 batches of Lonicerae Japonicae Flos dispensing granules of different standards (national standard/enterprise standard, NS/ES), and the anti-inflammatory potency of the above samples and half inhibitory concentration (IC₅₀) of the relevant monomer components were determined by using the cyclooxygenase-2 (COX-2) kit, and then the weight coefficient of anti-inflammatory potency of the above components was obtained by integrative analysis, and ECI was calculated through the integration of weight coefficients and effectors, and finally, the feasibility of the constructed ECI was verified by cellular experiments. Results The results of anti-inflammatory component content determination showed that there were large differences in the contents of nine individual components, of which there were also significant differences in the content of individual components between the NS batch (S1-S11) and the ES batch (S12-S22), for example, the content of chlorogenic acid in the NS batch (S8) was 3.3 times more than the content of this component in the ES batch (S16) (43.06 vs 12.98 mg/g). The overall stability of NS (S1-S11) samples was not much different from that of ES (S12-S22) samples (RSD_NS=211.55, RSD_ES=215.71). The bioefficacy assay passed the methodological examination, and the results showed that the anti-inflammatory potency and stability of the NS samples were better than those of the ES samples (potency_NS=100.28-106.35 U/mL, potency_ES=88.06-95.49 U/mL; RSD_NS=1.98, RSD_ES=2.56). The theoretical calculations of ECI showed that the results of the method were consistent with the experimental results, which could reflect the pharmacodynamic relationship of the Lonicerae Japonicae Flos dispensing granules samples (ECI_NS=10.82-14.11, ECI_ES=9.32-12.58; RSD_NS=9.51, RSD_ES=10.02). Correlation analysis showed that the better positive correlation between the content of single indicator components and anti-inflammatory potency was chlorogenic acid (r =0.644), followed by cymaroside (r =0.581), suggesting that these two components played a major role in anti-inflammatory activity. Conclusion This study establishes the quality evaluation method of Lonicerae Japonicae Flos dispensing granules based on ECI by taking anti-inflammation as an example. It can be applied to the quality consistency evaluation of multiple batches of Lonicerae Japonicae Flos dispensing granules with different standards (NS/ES), and provide scientific reference for the quality evaluation research and application of other Chinese medicine dispensing granules with similar anti-inflammatory effects.

河南省高校科技创新团队（23IRTSTHN026）；国家自然科学基金面上项目（82173993）；国家中医管理局中医药高质量发展协同创新转化工程自主立项项目（CXZH03）