健骨痛消丸中活性成分的检测及其抗凝血作用的研究

Determination of active components and study on anticoagulative action of Jiangutongxiao Pills

[目的]检测健骨消痛丸的活性物质及其抗凝血作用。[方法]采用高效液相色谱-示差折光检测法(HPLC-RID)对健骨痛消丸中松脂醇二葡萄糖苷(PDG)、京尼平、京尼平苷的含量进行定量测定。采用磁珠法测定血小板聚集率,分离血清利用半自动凝血仪测定活性成分及提取液凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)和纤维蛋白原(FIB),结果与对照组比较。[结果]HPLC-RID法测定PDG、京尼平和京尼平苷的浓度在一定浓度范围内线性关系良好。PDG的回归方程为Y=0.142X-0.0004(R2=0.9999),平均加样回收率为102.4%(RSD=3.9%);京尼平的回归方程为Y=0.003X+0.001(R2=0.9970),平均加样回收率为99.2%(RSD=2.8%);京尼平苷的回归方程为Y=0.011X+0.002(R2=0.9940),平均加样回收率为101.6%(RSD=3.2%)。采用该方法测定健骨痛消丸中PDG、京尼平和京尼平苷的含量分别为(3.52±0.085)mg/g、(32.6±1.6)μg/g和(221±9.5)μg/g。该方法测定结果与其他检测方法进行对比,无显著性差异;健骨痛消丸提取液PT、APTT、AT实验凝血时间显著大于灯盏花素,京尼平的PT活性最强,几种活性成分APTT活性均大于对照。[结论]所建立定量检测方法具有操作简便、准确可靠、抗干扰性强等特点,可有效监控健骨消痛丸的质量。

[Objective]To determine active components and study on anticoagulative action of Jiangutongxiao pills.[Methods]The contents of pinoresinol diglucoside(PDG),genipin and geniposide were determined by HPLC-RID.The magnetic bead method was used to determine the platelet aggregation rate.The serum was seperated and the semi-automatic coagulation instrument was used to determine the active ingredients and extracts thrombin time(APTT),prothrombin time(PT),thrombin time(TT)and fibrinogen(FIB).The result is compared with the control group.[Results]The results of quantitative analysis showed that PDG concentration was linear in the certain range,and the regression equation of PDG was Y=0.142X-0.0004(R2=0.9999),and the average recovery rate was 102.4%(RSD=3.9%);the regression equation of genipin was Y=0.011X+0.002(R2=0.9940),and the average recovery rate was 99.2%(RSD=2.8%);the regression equation of geniposide was Y=0.003X+0.001(R2=0.9970),and the average recovery rate was 101.6%(RSD=3.2%).The contents of PDG,genipin and geniposide were respectively(3.52±0.085)mg/g,(32.6±1.6)μg/g和(221±9.5)μg/g in Jiangutongxiao pills.The coagulation time of PT,APTT and AT was significantly longer than that of breviscapine;the activity of genipine was the strongest,and coagulation time of APTT of active components was higher than that of the control.[Conclusion]The established quantitative methods are simple,accurate and reliable,which can be effectively used for the quality control of Jiangutongxiao pills;Jiangutongxiao pills has the anticoagulative action.