公开(公告)号：US11692186B2

公开(公告)日：2023-07-04

申请号：US17389382

申请日：2021-07-30

申请人： Shandong University ,  Shandong Institute for Food and Drug Control ,  Avanc Pharmaceutical Co., Ltd

发明人： Ruiqing Xian ,  Feng Shi ,  Liping Gong ,  Lianli Chi ,  Qunye Zhang ,  Weijian Wang ,  Hongming Du ,  Congcong Wang ,  Baojian Hang ,  Fengshan Wang

IPC分类号： C07K5/00 ,  G01N30/72 ,  C12N9/74 ,  C12Q1/37 ,  G01N30/06

CPC分类号： C12N9/6429 ,  C12Q1/37 ,  G01N30/06 ,  G01N30/7206

摘要： The present invention provides a snake venom thrombin marker peptide of Agkistrodon Halys Pallas and an application of the snake venom thrombin-like enzyme in identifying species of Hemocoagulase for Injection. The application includes the following steps of: dissolving a to-be-detected sample and a reference substance of the marker peptide respectively to prepare a test solution and a reference solution, and conducting alkylation reduction on the test solution and the reference solution with dithiothreitol and iodoacetamide; after diluting products with an ammonium bicarbonate solution, adding enzyme for hydrolysis; and after enzymolysis is finished, conducting centrifugation at a high speed, and injecting a supernatan into a liquid chromatography-mass spectrometer for analysis. This method is simple, convenient and rapid, is strong in specificity, fills the gap in identifying the source of species of the snake venom thrombin-like enzyme of Agkistrodon Halys Pallas and improves the quality control level.

公开(公告)号：US11467141B2

公开(公告)日：2022-10-11

申请号：US17389322

申请日：2021-07-29

申请人： Shandong Institute for Food and Drug Control ,  Shandong University ,  Avanc Pharmaceutical Co., Ltd

发明人： Feng Shi ,  Ruiqing Xian ,  Liping Gong ,  Weijian Wang ,  Lianli Chi ,  Congcong Wang ,  Baojian Hang ,  Xunjie Zhang ,  Li Nie ,  Fengshan Wang

IPC分类号： G01N30/88 ,  G01N30/72 ,  G01N30/96

摘要： The present invention relates to the field of chemical analysis detection and application, in particular to a marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas and an application thereof. The amino acid sequence of the marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas is TLCAGVMEGGIDTCNR. Characterizing the source of species and a content of the SVTLEs in a to-be-detected sample by using the marker peptide includes the following steps of: pretreating the to-be-detected sample by trypsin through enzymolysis, and taking a supernatant of an enzymolysis liquid as a test solution; and injecting the test solution and a reference solution into a liquid chromatography-mass spectrometer, and selecting a qualitative ion pair and a quantitative ion pair for detecting the source of species and a content of the SVTLEs in the to-be-detected sample.

公开(公告)号：US20220034856A1

公开(公告)日：2022-02-03

申请号：US17389322

申请日：2021-07-29

申请人： Shandong Institute for Food and Drug Control ,  Shandong University ,  Avanc Pharmaceutical Co.,Ltd

发明人： Feng Shi ,  Ruiqing Xian ,  Liping Gong ,  Weijian Wang ,  Lianli Chi ,  Congcong Wang ,  Baojian Hang ,  Xunjie Zhang ,  Li Nie ,  Fengshan Wang

IPC分类号： G01N30/88 ,  G01N30/96 ,  G01N30/72

摘要： The present invention relates to the field of chemical analysis detection and application, in particular to a marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas and an application thereof. The amino acid sequence of the marker peptide of snake venom thrombin-like enzymes (SVTLEs) from Agkistrodon Halys Pallas is TLCAGVMEGGIDTCNR. Characterizing the source of species and a content of the SVTLEs in a to-be-detected sample by using the marker peptide includes the following steps of: pretreating the to-be-detected sample by trypsin through enzymolysis, and taking a supernatant of an enzymolysis liquid as a test solution; and injecting the test solution and a reference solution into a liquid chromatography-mass spectrometer, and selecting a qualitative ion pair and a quantitative ion pair for detecting the source of species and a content of the SVTLEs in the to-be-detected sample.

公开(公告)号：US20220034853A1

公开(公告)日：2022-02-03

申请号：US17389382

申请日：2021-07-30

申请人： Shandong Institute for Food and Drug Control ,  Shandong University ,  Avanc Pharmaceutical Co.,Ltd

发明人： Ruiqing Xian ,  Feng Shi ,  Liping Gong ,  Lianli Chi ,  Qunye Zhang ,  Weijian Wang ,  Hongming Du ,  Congcong Wang ,  Baojian Hang ,  Fengshan Wang

IPC分类号： G01N30/06 ,  G01N30/72 ,  C12N9/74

摘要： The present invention provides a snake venom thrombin marker peptide of Agkistrodon Halys Pallas and an application of the snake venom thrombin-like enzyme in identifying species of Hemocoagulase for Injection. The application includes the following steps of: dissolving a to-be-detected sample and a reference substance of the marker peptide respectively to prepare a test solution and a reference solution, and conducting alkylation reduction on the test solution and the reference solution with dithiothreitol and iodoacetamide; after diluting products with an ammonium bicarbonate solution, adding enzyme for hydrolysis; and after enzymolysis is finished, conducting centrifugation at a high speed, and injecting a supernatan into a liquid chromatography-mass spectrometer for analysis. This method is simple, convenient and rapid, is strong in specificity, fills the gap in identifying the source of species of the snake venom thrombin-like enzyme of Agkistrodon Halys Pallas and improves the quality control level.