Comparison of crude heparin pretreatment methods and identification of crude heparin from various species origins by quantitative PCR
10.3969/j.issn.1005-1678.2016.06.60
- VernacularTitle:肝素样品前处理方法比较及荧光定量PCR法鉴定肝素原料种属来源
- Author:
Chaodong ZHOU
;
Haijiao BAI
;
Zhesu HUANG
- Publication Type:Journal Article
- Keywords:
magnetic beads;
agrose gel electrophoresis;
heparinase;
quantitative PCR;
crude heparin;
species identification;
quality control
- From:
Chinese Journal of Biochemical Pharmaceutics
2016;36(6):197-199
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare magnetic beads kit,agrose gel recovery kit and heparinase I three methods to purify the micro DNA from crude heparin, then use q-PCR to identify the species origins and select the best method.Methods Using magnetic beads kit,agrose gel recovery kit and heparinase I to purify micro DNA from crude heparin and combined the porcine,bovine and ovine identification kits to identify the species origins and conformed the minimum detection limit of different percentage of ovine crude heparin in porcine crude heparin.Results Three pretreatment methods all can solve the pretreatment difficulties and we found that the haparinase was the best method; the minimum detection limit was 0.01%of ovine crude heparin in porcine crude heparin.Conclusion The heparinase method is the best pretreatment method and can successfully solve the pretreatment difficulties.Heparinase combine the porcine, bovine and ovine identification kits can identify the species origins from crude heparin.
