Phosphorothioated oligonucleotides are often used as nucleic acid medicines, because of their stability compared to ordinary oligonucleic acids. During the development and quality control of nucleic acid drugs that are expected as next-generation pharmaceuticals, ion-pair reversed phase mode is often used to analyze the oligonucleotides. However, deposition of the ion-pair reagent on the column can be a problem. In this application, an unpurified product of synthetic phosphorothioated oligo-DNA of 20 mer was analyzed using HILICpak VN-50 2D, a polymer-based HILIC column.
A good separation of oligomers were achieved without the use of ion-pair reagent under the HILIC mode. The gradient mode developed used 50 mM ammonium formate (pH9.5) and acetonitrile.
Sample : 1 µL
Synthesized phosphothioated oligo-DNA 20 mer (crude)
(A*T*A*C*C*G*A*T*T*A*A*G*C*G*A*A*G*T*T*T ; * means phosphorothioated position)
1.0 mg/mL (in H2O)
Column : Shodex HILICpak VN-50 2D (2.0 mm I.D. x 150 mm) Eluent : (A); 50 mM HCOONH4 aq. (pH9.5)/(B); CH3CN Linear gradient ; (B %) 64 to 58 % (0 to 10 min), 58 % (10 to 15 min),
58 to 64 % (15 to 15.01 min), 64 % (15.01 to 20 min)
Flow rate : 0.25 mL/min Detector : UV (260 nm) (small cell volume) Column temp. : 40 °C
Sample Name Index
Operation Manual / Certificate of Analysis
Operation Manuals and Certificate of Analysis / Inspection Certificate for the following products can be downloaded here.
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Applications
- High Sensitive Analysis of Synthetic Oligo-DNAs by LC/MS (Comparison between VN-50 1D and VN-50 2D)
- LC/UV/MS Analysis of Oligo-DNA (VN-50 2D)
- Analysis of Oligo-DNAs (VN-50 2D)
- Analysis of Various Oligonucleotides by LC/UV/MS (VN-50 2D)
- Analysis of 2'-OMe and 2'-MOE Modified Phosphorothioated Oligo-RNA by LC/UV/MS (VN-50 2D)
- Analysis of 10 - 50mer Oligo-DNAs (VN-50 2D)
- Analysis of Oligonucleotides and Their Impurities (1) Truncated Oligonucleotides (VN-50 2D)
- Analysis of Oligonucleotides and Their Impurities (2) Base Alteration (VN-50 2D)