HPLC Method for Adenosine Triphosphate, Cytidine Diphosphate on Newcrom B by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Adenosine Triphosphate, Cytidine Diphosphate .

Cytidine-5′-diphosphate is a nucleoside diphosphate with C₉H₁₅N₃O₁₁P₂ chemical formula. It is used by RNA and DNA to draw upon it for building blocks. It is a precursor to cytidine triphosphate.

Adenosine 5′-Triphosphate is a nucleotide with the chemical formula C₁₀H₁₆N₅O₁₃P_3. It is referred to as “energy currency” of the cell due to it’s importance in cellular energy metabolism. It is generated in the mitochondria and is used in numerous various cellular processes.

Adenosine Triphosphate, Cytidine Diphosphate can be retained and analyzed using the Newcrom B stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with a formic acid buffer. Detection is performed using UV.

Nucleotides are important biological molecules which serve as subunits of nucleic acids. They are composed of a five-carbon sugar, a nitrogenous base, and at least one phosphate group. Nucleotides cannot be retained by reverse-phase chromatography without an ion-pairing reagent due to their highly polar nature. Primesep SB is capable of retaining and separating nine nucleotides. Primesep SB is a reverse-phase column with strong embedded basic ion-pairing groups.

Cytidine phosphates are nucleotides. Nucleotides contain a nucleobase, five-carbon sugar and one or three phosphate groups. All three cytidines are very polar and cannot be retained by reversed-phase chromatography without a ion-pairing reagent. Mixed-mode Primesep SB column easily allows to retain and separate mono-, di-, and tri-phospates by anion-exchange mechanism. Other nucleotides can be separated as well by this approach. Nucleotides in biofluids can be analyzed directly on this column without any significant sample preparation. Most of the proteins from biological fluids will be repelled from the surface of silica gel. Method is compatible with UV, ELSD, CAD and LC/MS.