[关键词]
[摘要]
目的 建立以抗血小板聚集活性为指标的生物检定法,评价川芎药材、饮片及含川芎中成药的质量。方法 川芎加水回流定量提取,以提取物为供试品体外测定抗血小板聚集活性。家兔心脏取血,制备富血小板血浆(PRP),用腺苷-5'-二磷酸钠盐诱导血小板聚集,以血小板抑制率为抗血小板聚集活性指标,用阿魏酸钠标定川芎提取物的抗血小板聚集活性。根据量反应平行线法(2.2)法计算川芎提取物的抗血小板聚集活性;结合川芎水提物的提取率,计算川芎样品抗血小板聚集活性。并测定了8份川芎药材、饮片及中成药样品的抗血小板聚集活性,验证方法的可行性。结果 阿魏酸钠和川芎提取物均具有显著抗血小板聚集活性(P<0.01),并且可靠性检验结果成立。阿魏酸钠在给药质量浓度15~60 mg/mL内与其血小板聚集抑制率呈良好的线性关系(r=0.993 4,n=4)。供试品重复测定抗血小板聚集活性的RSD值为3.43%(n=6),可信限率为23.90%(n=6)。不同川芎样品抗血小板聚集活性不同,4份川芎药材抗血小板聚集活性的生物效价分别为3.183、2.068、1.957和1.931 U/g,川芎饮片及川芎酒炙饮片抗血小板聚集活性的生物效价分别为1.304、1.021 U/g,速效救心丸和乐脉颗粒抗血小板聚集活性的生物效价分别为0.506、0.919 U/g。结论 建立的方法可准确测定川芎药材、饮片及含川芎中成药的抗血小板聚集活性,可用来评价川芎产品的质量。
[Key word]
[Abstract]
Objective To develop a bioassay method to quantify the antiplatelet aggregation bioactivity (AAB) of crude Chuanxiong Rhizoma, decotion pieces, and its Chinese patent medicine for quality assessment. Methods Chuanxiong Rhizoma sample was extracted in water by reflux. The level of AAB in extract was quantified in vitro. The blood was taken from the heart of rabbit. The platelet aggregating in platelet-rich plasma was induced by adenosine-5'-diphosphate disodium salt. The ratio of platelet inhibition was chosen as AAB marker. Sodium ferulate was a reference. The amount of AAB in aqueous extract was quantified by the Amount reaction of parallel line analysis (2.2) method. The AAB data of herbal sample was calculated by combining the AAB of extract with its extraction rate. Moreover, AAB amounts were quantified in the eight samples including crude Chuanxiong Rhizoma, decoction pieces, and Chinese patent medicines to verify this developed method. Results Both sodium ferulate and Chuanxiong extract showed significant AAB (P< 0.01). The reliability test for quantifying AAB in solidum ferulate and Chuanxiong Rhizoma extract was passed through, and the measured value was valid. The correlation coefficient was 0.993 4 (n=4) between the amounts of solidum ferulate in the concentration range of 15-60 mg/mL and their ratios of platelet inhibition. The RSD for the amounts of AAB was 3.43% (n=6) by six replicated tests with the confidence limit rate of 23.90% (n=6). The AAB amounts were significantly different among tested samples, i.e. 3.183, 2.068, 1.957, and 1.931 U/g for four Chuanxiong Rhizoma crude samples, 1.304 and 1.021 U/g for Chuanxiong Rhizoma decotion pieces and processed slice with yellow wine, 0.506 and 0.919 U/g for Suxiao Jiuxin Pills and Lemai Granule. Conclusion The developed method can accurately quantify the level of AAB in Chuanxiong Rhizoma crude, decoction pieces, and Chinese patent medicines, which can be used to assess the product quality of Chuanxiong Rhizoma.
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[基金项目]
四川省重点研发项目(2017SZ0156);四川省教育厅资助科研项目(17za0148)