title: New approach to the determination phosphorothioate oligonucleotides by ultra high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry.
abstract: This text presents a novel method for the separation and detection of phosphorothioate oligonucleotides with the use of ion pair ultra high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry The research showed that hexafluoroisopropanol/triethylamine based mobile phases may be successfully used when liquid chromatography is coupled with such elemental detection. However, the concentration of both HFIP and TEA influences the final result. The lower concentration of HFIP, the lower the background in ICP-MS and the greater the sensitivity. The method applied for the analysis of serum samples was based on high resolution inductively coupled plasma mass spectrometry. Utilization of this method allows determination of fifty times lower quantity of phosphorothioate oligonucleotides than in the case of quadrupole mass analyzer. Monitoring of (31)P may be used to quantify these compounds at the level of 80 μg L(-1), while simultaneous determination of sulfur is very useful for qualitative analysis. Moreover, the results presented in this paper demonstrate the practical applicability of coupling LC with ICP-MS in determining phosphorothioate oligonucleotides and their metabolites in serum within 7 min with a very good sensitivity. The method was linear in the concentration range between 0.2 and 3 mg L(-1). The limit of detection was in the range of 0.07 and 0.13 mg L(-1). Accuracy varied with concentration, but was in the range of 3%.
authors.affiliation: Chair of the Environmental Chemistry & Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., PL-87 100 Torun, Poland. Electronic address: kowalska@chem.umk.pl.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Studzińska
authors.affiliation: CNRS/UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR 5254, Hélioparc, 2 av. Pr. Angot, F-64053 Pau, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Mounicou
authors.affiliation: CNRS/UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR 5254, Hélioparc, 2 av. Pr. Angot, F-64053 Pau, France.
authors.forename: Joanna
authors.initials: J
authors.lastname: Szpunar
authors.affiliation: CNRS/UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR 5254, Hélioparc, 2 av. Pr. Angot, F-64053 Pau, France.
authors.forename: Ryszard
authors.initials: R
authors.lastname: Łobiński
authors.affiliation: Chair of the Environmental Chemistry & Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., PL-87 100 Torun, Poland.
authors.forename: Bogusław
authors.initials: B
authors.lastname: Buszewski
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D008970:Molecular Weight; D054735:Phosphorothioate Oligonucleotides / Q000032:analysis* / Q000097:blood / Q000737:chemistry* / Q000235:genetics; D010758:Phosphorus / Q000032:analysis; D013455:Sulfur / Q000032:analysis
chemical_list: D054735:Phosphorothioate Oligonucleotides; D010758:Phosphorus; D013455:Sulfur
keywords: Hexafluoroisopropanol/triethylamine; Inductively coupled plasma mass spectrometry; Ion pair chromatography; Phosphorothioate oligonucleotides; Serum