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：US20220033873A1

公开(公告)日：2022-02-03

申请号：US17389384

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： Feng Shi ,  Liping Gong ,  Ruiqing Xian ,  Lianli Chi ,  Weijian Wang ,  Baojian Hang ,  Congcong Wang ,  Pengfei You ,  Hongming Du

IPC分类号： C12Q1/37 ,  G01N30/72 ,  G01N30/86 ,  G01N30/06

摘要： The present invention relates to the technical field of chemical analysis and quantitative detection, in particular to a quantitative detection method for snake venom thrombin-like enzyme (SVTLE) from Agkistrodon halys pallas. The quantitative detection method for the SVTLE includes the following steps of taking a reference substance of marker peptide for the SVTLE from Agkistrodon halys pallas with an amino acid sequence of LDSPVSNSAHIAPLSLPSSAPSVGSVCR, and preparing a series of reference solutions with different concentrations; adding the reference solutions in test solutions respectively for enzymolysis, and then taking a supernatant after enzymolysis as a series of solutions to be detected; and adding the solutions to be detected in a liquid chromatogram-mass spectrometer, and then selecting a qualitative ion pair and a quantitative ion pair to detect contents of marker peptide in the solutions to be detected.

公开(公告)号：US11584948B2

公开(公告)日：2023-02-21

申请号：US17389384

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： Feng Shi ,  Liping Gong ,  Ruiqing Xian ,  Lianli Chi ,  Weijian Wang ,  Baojian Hang ,  Congcong Wang ,  Pengfei You ,  Hongming Du

IPC分类号： C12Q1/37 ,  G01N30/06 ,  G01N30/72 ,  G01N30/86 ,  G01N30/02

摘要： The present invention relates to the technical field of chemical analysis and quantitative detection, in particular to a quantitative detection method for snake venom thrombin-like enzyme (SVTLE) from Agkistrodon halys pallas. The quantitative detection method for the SVTLE includes the following steps of taking a reference substance of marker peptide for the SVTLE from Agkistrodon halys pallas with an amino acid sequence of LDSPVSNSAHIAPLSLPSSAPSVGSVCR, and preparing a series of reference solutions with different concentrations; adding the reference solutions in test solutions respectively for enzymolysis, and then taking a supernatant after enzymolysis as a series of solutions to be detected; and adding the solutions to be detected in a liquid chromatogram-mass spectrometer, and then selecting a qualitative ion pair and a quantitative ion pair to detect contents of marker peptide in the solutions to be detected.

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.