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建立红细胞凝集反应过程的微量量热检测方法,并用于评价不同产地板蓝根的活性差异。采用微量量热法,检测阳性参照物(植物凝集素,PHA)及8批不同产地板蓝根对20%家兔血红细胞凝集反应过程,记录凝集反应热动力学曲线,提取反应动力学参数,计算凝集效价,对结果进行主成分分析和聚类分析,并以微量板法对结果进行验证。结果表明,板蓝根对家兔血红细胞凝集反应为放热反应,反应动力学参数如反应速率常数(k:0.039～73.6min-1)、最大反应功率(Pmax:-1140.2～988.2μW)及反应焓变(Hi:-529.9～717.9μJ)等动力学参数与板蓝根浓度(0.2～1.0g·mL-1)呈良好的线性关系(r>0.97);主成分分析结果显示参数Pmax(531～1335μW)及Ht(585.2～989.2μJ)能够客观表征不同产地板蓝根样品凝集活性差异,与微量板法测定结果(不同产地板蓝根凝集滴度分别为3～11)基本一致。根据效价值可区分GAP产区、一般主产区和普通散种区样品,同时凝集反应曲线具有较好的特征性和产地专属性,能为板蓝根品质评价提供更加全面的综合信息。所建立的板蓝根血红细胞凝集反应的微量量热检测方法灵敏可靠,可作为凝集反应定量在线检测手段,并为板蓝根及其他具凝集活性类中药的品质评价提供新的技术参考。
To establish the method of micro-calorimetry for the hemagglutination reaction and to evaluate the differences of the activity of Radix Isatidis in different areas. The microcalorimetry was used to detect the positive reference (phytohemagglutinin, PHA) and 8 batches of Radix Isatidis on the hemagglutination reaction of 20% rabbits. The thermokinetic curve of agglutination reaction was recorded and the kinetic parameters were extracted to calculate the agglutination Titer, the results of principal component analysis and cluster analysis, and micro-plate method to verify the results. The results showed that Radix Isatidis was an exothermic reaction on rabbit hemagglutination. The reaction kinetic parameters such as reaction rate constant (k: 0.039 ~ 73.6min-1), maximum reaction power (Pmax: -1140.2 ~ 988.2μW) and reaction enthalpy (P <0.05), and the kinetic parameters (Hi: -529.9 ~ 717.9μJ) were linear with the concentrations of isatis root (0.2 ~ 1.0g · mL-1) And Ht (585.2 ~ 989.2μJ) were able to objectively characterize the agglutination activity difference of Radix Isatidis samples from different areas. The results were in good agreement with the results of microtiter plate assay (titer of three to 11 in Radix Isatidis from different habitats). According to the value of the value can distinguish GAP producing areas, the general main producing areas and ordinary litter area samples, while the agglutination reaction curve has good characteristics and originality, can provide a more comprehensive and comprehensive information for the quality evaluation of Banlangen. The established microtrophic detection method for the hemagglutination of the Radix isatidis is sensitive and reliable and can be used as a quantitative online detection method for the agglutination reaction and provides a new technical reference for the quality evaluation of Radix Isatidis and other agglutinative active Chinese medicines.