Development of a solid-phase extraction/gas chromatographic-mass spectrometric method for quantification of succinic acid in nucleoside derivatives for oligonucleotide synthesis.
A solid-phase extraction (SPE)/gas chromatographic-mass spectrometric (GC-MS) method was developed for analysing residual succinic acid in nucleoside derivatives to be used in oligonucleotide synthesis. Use of a SPE protocol, enabled most of the derivatives to be trapped, thereby creating eluates enriched in succinic acid. GC-MS was used to quantify the amount of residual succinic acid in four different nucleoside preparations, with succinate concentrations varying from 0.18 to 0.24% (w/w). The within-day repeatability of the method was found to be 1.25% RSD. A linear relationship was observed between the amount of succinic acid in the sample and the GC-MS peak area, with a correlation coefficient of 0.9997 in the concentration interval 0.05-2.5% (w/w). Recoveries were measured by the addition of internal standards to working solutions and varied between 99.8 and 102.6%.
Stenholm
Ake
A
Process Analytical Chemistry, Amersham Biosciences, Björkgatan 30, S-75184 Uppsala, Sweden. ake.stenholm@amersham.com
Drevin
Ingrid
I
Lundgren
Michael
M
D002853:Chromatography, Liquid; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D009705:Nucleosides / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D019802:Succinic Acid / Q000032:analysis*
D009705:Nucleosides; D009841:Oligonucleotides; D019802:Succinic Acid