title: Purification and characterization of saxitoxin from Mytilus chilensis of southern Chile.
abstract: In the current communication we describe an innovative method to purify saxitoxin (STX), a toxin presents in contaminated muscle of Mylitus chilensis extracted in the southern part of Chile, using a liquid chromatographic methodology based on ionic pairs. The STX was extracted using HCl and treated with ammonium sulfate following a treatment with trichloroacetic acid and hexane/diethyl ether (97/3). The samples were analyzed by a semi-preparative HPLC in order to collect pure fractions of STX and these fractions were eluted in solid-phase cationic interchange SCX extraction columns. The purified STX was stable and homogeneous and its identity was confirmed by LC-MS-MS, which demonstrated a high quality purification of STX, without presence of analogs such as neosaxitoxin (Neo) and decarbamoyl saxitoxin (dcSTX). The STX biological activity was analyzed in a bioassay in mice model and compared to the standard STX produced by the FDA and no significant differences were observed.
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: D P
authors.initials: DP
authors.lastname: Rubio
authors.affiliation: Sección de Química y Alimentos, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: L G
authors.initials: LG
authors.lastname: Roa
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: D A
authors.initials: DA
authors.lastname: Soto
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: F J
authors.initials: FJ
authors.lastname: Velasquez
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: N A
authors.initials: NA
authors.lastname: Gregorcic
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile; Millennium Institute on Immunology and Immunotherapy, Chile; Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Chile.
authors.forename: J A
authors.initials: JA
authors.lastname: Soto
authors.affiliation: Sección Microbiología de Alimentos, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile.
authors.forename: M C
authors.initials: MC
authors.lastname: Martinez
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, Chile; Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Chile.
authors.forename: A M
authors.initials: AM
authors.lastname: Kalergis
authors.affiliation: Sección de Biotecnología, Departamento de Salud Ambiental, Instituto de Salud Púbica de Chile, Chile; Millennium Institute on Immunology and Immunotherapy, Chile. Electronic address: avasquez@ispch.cl.
authors.forename: A E
authors.initials: AE
authors.lastname: Vasquez
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D002677:Chile; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid; D006851:Hydrochloric Acid / Q000737:chemistry; D051379:Mice; D049878:Mytilus / Q000737:chemistry*; D012530:Saxitoxin / Q000737:chemistry / Q000302:isolation & purification*; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry
chemical_list: D012530:Saxitoxin; D006851:Hydrochloric Acid; D000645:Ammonium Sulfate
keywords: HPLC; Harmful algal blooms; Mylitus chilensis; Paralytic shellfish poison; Saxitoxin