目的 建立一测多评（QAMS）结合化学模式识别与加权逼近理想排序（TOPSIS）法的不同产地小叶三点金Desmodiummicrophyllum总黄酮（DMTF）质量综合评价方法，初步探讨其抗炎活性。方法 采用Thermo C₁₈色谱柱（250 mm× 4.6 mm,5 μm） ，流动相乙腈-0.1%甲酸梯度洗脱，体积流量0.8 mL·min^-1，柱温20℃，检测波长300、350 nm，进样量10 μL。以异荭草苷为内标，建立其与夏佛塔苷、荭草苷、异夏佛塔苷、木犀草素的相对校正因子，测定待测组分的含量，采用外标法（ESM）测定DMTF中这5种成分的含量，并比较两种方法的差异，以验证QAMS法的准确性和可靠性。结合化学模式识别和TOPSIS分析，评价不同产地DMTF质量差异性。采用酶联免疫吸附法（ELISA），观察ig给予临床等效剂量DMTF后采集的大鼠含药血清对脂多糖（LPS，1 μg·mL^-1）诱导的巨噬细胞（RAW 264.7）释放的白细胞介素（IL）-1β、IL-6含量的影响。结果 以异荭草苷为内标，夏佛塔苷、荭草苷、异夏佛塔苷、木犀草素的相对校正因子分别为1.498、1.050、1.522、0.653，5种成分线性关系良好（r≥0.999 0），QAMS法与ESM所得结果无显著性差异。化学模式识别法将10批样品分为2类，荭草苷、异荭草苷、夏佛塔苷和异夏佛塔苷是影响DMTF质量的主要差异性成分。加权TOPSIS法显示10批DMTF欧氏贴近度为0.123 3～0.952 0，提示不同产地样品存在一定差异，质量优劣排序和分类与化学模式识别聚类结果基本一致。抗炎活性评价结果显示，与对照组（10% FBS）相比，模型组IL-1β和IL-6水平显著升高（P＜0.01） ；与模型组相比，10%含药血清和阳性药地塞米松（1 μg·mL^-1 LPS＋10 μg·mL^-1）均能显著降低IL-1β和IL-6水平（P＜0.01）。结论 所建立的QAMS结合化学模式识别及加权TOPSIS法可用于评价不同产地DMTF质量差异性； DMTF具有较好的体外抗炎活性，为小叶三点金质量标准提升提供参考。

Objective To establish a comprehensive quality evaluation method for different origins of total flavonoids of Desmodium microphyllum (DMTF) by integrating QAMS, chemical pattern recognition and weighted TOPSIS, and to preliminarily explore its antiinflammatory activity. Methods Using a Thermo C₁₈ chromatographic column (250 mm × 4.6 mm, 5 μm), the mobile phase was acetonitrile-0.1% formic acid with a gradient elution, with a volume flow rate of 0.8 mL·min^-1, a column temperature of 20 ℃, and a detection wavelength of 300 and 350 nm. Using isoorientin as the internal standard and establishing its relative correction factor with schaftoside, orientin, isoschaftoside, and luteolin. The content of the analyte components was determined, and the content of these five components in DMTF was determined by the external standard method (ESM), and the differences between the two methods were compared to verify the accuracy and reliability of the QAMS method. Combined with chemical pattern recognition and TOPSIS analysis, the quality differences of DMTF from different origins were evaluated. The effect of drug-containing serum collected from rats after intragastric administration of the clinical equivalent dose of DMTF on the release of interleukin (IL)-1β and IL-6 by lipopolysaccharide (LPS, 1 μg·mL^-1)-induced macrophages (RAW264.7) was observed by enzyme-linked immunosorbent assay (ELISA). Results Using isoorientin as the internal standard, the relative correction factors of schaftoside, orientin, isoschaftoside, and luteolin were 1.498, 1.050, 1.522, 0.653, respectively. The linear relationship of the five components was good (r ≥ 0.999 0). There was no significant difference between the QAMS method and the ESM results. The chemical pattern recognition method classified 10 batches of samples into two categories, and orientin, isoorientin, schaftoside, and isoschaftoside were the main differential components affecting the quality of DMTF. The weighted TOPSIS method showed that the euclidean proximity degree of the 10 batches of DMTF was 0.123 3－ 0.952 0, suggesting that there were certain differences in the samples from different origins, and the ranking and classification of quality were basically consistent with the clustering results of chemical pattern recognition. The anti-inflammatory activity evaluation results showed that compared with the control group (10% FBS), the IL-1β and IL-6 levels in the model group were significantly increased (P < 0.01); compared with the model group, both the 10% drug-containing serum group and the positive drug group (1 μg·mL^-1 LPS + 10 μg·mL^-1 dexamethasone) could significantly reduce the levels of IL-1β and IL-6 (P < 0.01). Conclusion The QAMS combined with chemical pattern recognition and weighted TOPSIS method can be used to evaluate the quality differences of DMTF from different origins; DMTF has good in vitro anti-inflammatory activity, providing a reference for the improvement of the quality standard of D. microphyllum.

R284.1

中央引导地方科技发展资金资助项目（202201002）；广西壮瑶药重点实验室开放课题（GXZYZZ202010）；广西中医药大学第三批“岐黄工程”高层次人才团队培育项目（202406）；广西中医药管理局项目（GZZJ202029，GZZJ202030）；广西中医药重点研究室项目（桂中医药科教发［2023］9号）；国家中医药管理局全国名老中医药专家传承工作室建设项目-黄瑞松全国名老中医药专家传承工作室（国中医药人教函［2022］75号）