On-column electroextraction and separation of antisense oligonucleotides in human plasma by capillary gel electrophoresis

A combined solid phase extraction/capillary gel electrophoresis method for the determination of phosphorothioate oligodeoxynucleotides in biological fluids, tissues and feces

“…Furthermore, this assay could be used to extract PS-ODNs of different base lengths in various biological samples. The LLOQ and LOD of the current method are similar to those of previously reported CGE methods [7,10], but not as low as the LC-MS/MS methods or hybridization-based ELISAs. Therefore, this assay constitutes an alternative for quantitating oligodeoxynucleotides from biological samples.…”

“…To date, a variety of bioanalytical methods, such as capillary gel electrophoresis (CGE) [6][7][8][9][10], CGE-MS [11,12], capillary zone electrophoresis (CZE) [13], high-performance liquid chromatograph (HPLC) [14], liquid chromatography-mass spectrometry (LC-MS) [15][16][17], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [18][19][20][21] and hybridization-based enzyme-linked immunosorbent assay (ELISA) [22], have been developed and used in quantitation of PS-ODNs and metabolites. Among these analytical techniques, CGE is widely employed in preclinical and clinical studies for ASOs and allows the separation of the parent compound from chain-shortened metabolites at good resolutions and reasonable sensitivities.…”

“…However, one common problem in developing CGE methods for quantitating PS-ODNs is the need for rather extensive sample clean-up for the minimization of endogenous interference with ASOs during electrokinetic injection. Typical plasma and urine sample clean-up procedures for CGE analysis involve two-step SPE followed by additional desalting via dialysis, while the sample preparation from tissue and cellular fractions normally requires liquid/liquid extraction prior to analysis, and additional clean-up with solid-phase extraction for CGE analysis [6][7][8][9][10][11][12][13][14][15]23,24]. Recently, many attempts have been made to simplify and speed up these laborious sample clean-up procedures.…”

Electric field‐enhanced transport across phase boundaries and membranes and its potential use in sample pretreatment for bioanalysis

“…Method for the simultaneous EE and injection of phosphorothioate oligonucleotides directly from human plasma into a fused-silica capillary for subsequent analysis by CGE was also presented [84]. The capillary was filled with dextran solution, then with a short plug of acidified water and subsequently was dipped into the human plasma sample for simultaneous EE and injection.…”

Analysis of nucleic acid constituents by on‐line capillary electrophoresis‐mass spectrometry

“…In CGE, it is necessary to use coated capillaries to reduce the EOF to prevent the migration of the gel. When gel-filled capillaries are used for the determination of negatively charged analytes, the power supply of the CE system must therefore be in reversed-polarity mode, i.e., with the cathode on the injection side and the anode on the detection side (see Table 2) [8,9,94,110,[125][126][127].…”