Polyanionic acids to improve recovery and minimize system loss
The present technology relates to a method of separating a sample comprising oligonucleotides. The method includes injecting a polyphosphonic acid at a concentration of between about 0.01 M to about 1 M into the sample comprising oligonucleotides. The method also includes flowing the sample and polyphosphonic acid through a liquid chromatography column and separating the oligonucleotides.
1. A method of performing solid phase extraction comprising:
injecting a sample comprising oligonucleotides into a solid phase extraction cartridge comprising a stationary phase, wherein the oligonucleotides are retained by the stationary phase;
washing the solid phase extraction cartridge with a polyphosphonic acid at a concentration of between about 0.01 M to about 1 M to elute the oligonucleotides from the solid phase extraction cartridge.
2. The method of claim 1 , wherein the concentration of polyphosphonic acid is between about 0.01 M to about 0.1 M.
3. The method of claim 2 , wherein the concentration of polyphosphonic acid is between about 20 mM to about 50 mM.
4. The method of claim 3 , wherein the concentration of polyphosphonic acid is between about 30 mM to about 40 mM.
5. The method of claim 1 , wherein between about 1 pg to about 1 mg of the polyphosphonic acid is injected into the sample.
6. The method of claim 1 , wherein the polyphosphonic acid is etidronic acid.
7. The method of claim 1 , wherein the polyphosphonic acid is nitrilotri(methylphosphonic acid).
8. The method of claim 1 , wherein walls of the solid phase extraction cartridge are formed of plastic.
9. The method of claim 1 , wherein the sample comprises phosphorylated oligonucleotides.
10. The method of claim 9 , wherein the phosphorylated oligonucleotides comprise nucleotides and at least one nucleotide is adenosine triphosphate.
11. The method of claim 1 , wherein the polyphosphonic acid is etidronic acid at pH of 8.5 and the oligonucleotide comprises nucleotides and at least one nucleotide is adenosine triphosphate.
12. The method of claim 1 , wherein the polyphosphonic acid is a bisphosphonic acid.
13. The method of claim 12 , wherein the bisphosphonic acid is selected from the group consisting of clodronic acid, pamidronic acid, alendronic acid, neridronic acid, and olpadronic acid.
14. The method of claim 1 , wherein the solid phase extraction cartridge is a packed syringe.
15. The method of claim 1 , wherein the solid phase extraction cartridge is a well plate.