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首页> 外文期刊>RSC Advances >Rapid magnetic solid-phase extraction combined with ultra-high performance liquid chromatography and quadrupole-time-of-flight mass spectrometry for analysis of thrombin binders from a crude extract and injection of Erigeron breviscapus
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Rapid magnetic solid-phase extraction combined with ultra-high performance liquid chromatography and quadrupole-time-of-flight mass spectrometry for analysis of thrombin binders from a crude extract and injection of Erigeron breviscapus

机译：快速磁相萃取结合超高效液相色谱法和四极 - 飞行时间质谱法，用于分析来自原油提取物和Erigeron Breviscapus的克林蛋白粘合剂的分析

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Thrombin, which acts as a serine protease that converts soluble fibrinogen into insoluble strands of fibrin, plays crucial roles in the blood coagulation pathway. Inhibition of thrombin leading to the decrease of thrombus formation may be used for treating cerebral ischemia and infarction. In the present study, a new approach of thrombin immobilized magnetic nanoparticles coupled with ultra-high performance liquid chromatography and quadrupole-time-of-flight mass spectrometry was established for rapidly extraction of thrombin binders from a crude extract of Erigeron breviscapus and its injection. Scutellarin, a model inhibitor of thrombin, was chosen to interrogate the optimal extraction conditions, including wash times, ion strength, temperature, pH, and incubation time. Meanwhile, the specificity of the approach was investigated by using denatured thrombin and the inactive compound specnuezhenide. The optimal screening conditions were as follows: wash times: 4, wash solvent: 50% acetonitrile–water, incubation time: 70 min, temperature: 37 °C, ion strength: 10 mM, pH: 6.8. Moreover, the linearity, accuracy, precision, and matrix effect of the approach were well validated. Finally, thrombin binders were extracted from the crude extract and injection of Erigeron breviscapus by applying the established approach, and were subsequently sent to be validated by a thrombin inhibitory assay. Eleven thrombin inhibitors were identified, and ten of which, including erigoster B, scutellarin A, erigeroside, neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, luteolin 7- O -β- D -glucuronide, 4,5-di- O -caffeoylquinic acid, 3,4-di- O -caffeoylquinic acid and 3,5-di- O -caffeoylquinic acid, were reported for the first time.
机译：凝血酶，其作为丝氨酸蛋白酶，将可溶性纤维蛋白原转化成纤维蛋白的不溶链，在血液凝固途径中起着至关重要的作用。血栓抑制导致血栓形成减少可用于治疗脑缺血和梗塞。在本研究中，建立了一种新的凝血酶固定化磁性纳米粒子与超高效液相色谱和四极掺杂时间质谱法的新方法，从Erigeron Breviscapus的粗提物中快速提取凝血酶粘合剂及其注射。选择凝血酶模型抑制剂的Scutellarin以询问最佳提取条件，包括洗涤时间，离子强度，温度，pH和孵育时间。同时，通过使用变性凝血酶和惰性化合物甲烯烯来研究该方法的特异性。最佳筛查条件如下：洗涤时间：4，洗涤溶剂：50％乙腈 - 水，孵育时间：70分钟，温度：37℃，离子强度：10 mm，pH：6.8。此外，该方法的线性，准确性，精度和矩阵效应良好验证。最后，通过施加建立的方法从粗提取物和注射埃里耳孔景观中提取凝血酶粘合剂，随后通过凝血酶抑制测定来验证。鉴定了11个凝血酶抑制剂，其中十分之一，其中包括Esigoster B，Scutellarin A，Erigeroside，新泛酸，绿原酸，4-咖啡酸，叶英蛋白7-O-β-D-葡糖苷，4,5-二℃ - 第一次报道咖啡酰基丙酸，3,4-二-CaffeoylquInic酸和3,5-二丁蛋白酸酸。

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