title: Isolation and purification of monosialotetrahexosylgangliosides from pig brain by extraction and liquid chromatography.
abstract: Monosialotetrahexosylganglioside (GM1), one of glycosphingolipids containing sialic acid, plays particularly important role in fighting against paralysis, dementia and other diseases caused by brain and nerve damage. In this work, a simple and highly efficient method with high yield was developed for isolation and purification of GM1 from pig brain. The method consisted of an extraction by chloroform-methanol-water and a two-step chromatographic separation by DEAE-Sepharose Fast Flow anion-exchange medium and Sephacryl S-100 HR size-exclusion medium. The purified GM1 was proved to be homogeneous and had a purity of >98.0% by high-performance anion-exchange and size-exclusion chromatography. The molecular weight was 30.0 kDa by high-performance size-exclusion chromatography and 1546.9 Da by electrospray ionization mass spectrometry. The chromogenic reaction by resorcinol-hydrochloric acid solution indicated that the purified GM1 showed a specific chromogenic reaction of sialic acid. Through this isolation and purification program, ~1.0 mg of pure GM1 could be captured from 500 g wet pig brain tissue and the yield of GM1 was around 0.022%, which was higher than the yields by other methods. The method may provide an alternative for isolation and purification of GM1 in other biological tissues.
authors.affiliation: College of Life Science, Northwest University, Xi'an, 710069, China.
authors.forename: Liujiao
authors.initials: L
authors.lastname: Bian
authors.affiliation: College of Life Science, Northwest University, Xi'an, 710069, China.|Weapon Industry 521 Hospital, Xi'an, 710065, China.
authors.forename: Jianting
authors.initials: J
authors.lastname: Yang
authors.affiliation: College of Life Science, Northwest University, Xi'an, 710069, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000096:Acetone / Q000737:chemistry; D000818:Animals; D001923:Brain Chemistry; D011232:Chemical Precipitation; D002725:Chloroform / Q000737:chemistry; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D005677:G(M1) Ganglioside / Q000737:chemistry* / Q000302:isolation & purification*; D000432:Methanol / Q000737:chemistry; D008970:Molecular Weight; D012118:Resorcinols / Q000737:chemistry; D012685:Sepharose; D012794:Sialic Acids / Q000737:chemistry; D034421:Sus scrofa
chemical_list: D012118:Resorcinols; D012794:Sialic Acids; D000096:Acetone; D005677:G(M1) Ganglioside; D002725:Chloroform; D012685:Sepharose; D000432:Methanol; C031389:resorcinol
keywords: extraction; isolation and purification; liquid chromatography; monosialotetrahexosylganglioside; pig brain