In order to develop an affinity HPLC method for screening direct thrombin inhibitors from Traditional Chinese Medicine (TCM), thrombin was immobilized on the glutaraldehyde-modified amino silica gel and was used as thrombin stationary phase. A thrombin affinity column (TAC) was made by packing the thrombin stationary phase into a bare column (2.0 * 1.0 mm, i.d.). The direct thrombin inhibitors could be screened through this TAC column. For the purpose of improvement of the discovery efficiency, a TAC-HPLC-MS/MS system was used to screen thrombin inhibitors from Radix Salviae Miltiorrhiae (RSM), a famous traditional Chinese medicine. After optimization of all the conditions, cryptotanshinone (Cry), dihydrotanshinone I (Dih-I) and tanshinone IIA (Tan-IIA) were screened out and identified as potential active components. The anticoagulant effects of these three compounds were tested by anticoagulant experiments in vitro. Furthermore, the interaction of three compounds with thrombin was studied by molecular docking. The result shows they have the potential to be used as preventive drugs. In short, this method can be used to screen anticoagulant drugs from traditional Chinese medicine, which provides convenience for screening anticoagulant drugs.

中文翻译：

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建立凝血酶亲和柱（TAC）-HPLC-MS / MS方法从丹参中直接筛选凝血酶抑制剂。

为了开发一种亲和力高效液相色谱法从中药（TCM）中筛选直接凝血酶抑制剂，将凝血酶固定在戊二醛修饰的氨基硅胶上，并用作凝血酶固定相。凝血酶亲和柱（TAC）是通过将凝血酶固定相装填到裸柱（2.0 * 1.0 mm，id）中而制得的。直接凝血酶抑制剂可通过该TAC色谱柱进行筛选。为了提高发现效率，使用TAC-HPLC-MS / MS系统从著名的中药丹参（RSM）中筛选凝血酶抑制剂。在优化所有条件后，筛选出隐丹参酮（Cry），二氢丹参酮I（Dih-I）和丹参酮IIA（Tan-IIA），并将其鉴定为潜在的活性成分。通过体外抗凝实验测试了这三种化合物的抗凝作用。此外，通过分子对接研究了三种化合物与凝血酶的相互作用。结果表明它们具有用作预防药物的潜力。简而言之，该方法可用于从中药中筛选抗凝药物，为筛选抗凝药物提供了便利。