G1007615

LC/MS Analysis of Nucleotides on SeQuant^® ZIC-cHILIC

application for LC-MS

Properties

• technique(s): LC/MS: suitable
• test parameters: column: SeQuant^® ZIC-cHILIC, 10 cm x 2.1 mm, 3.0 μm particles (1.50657); mobile phase: [A] acetonitrile; [B] 50 mM ammonium acetate, pH 5.0 with acetic acid in water; gradient: 26 to 27% B in 10 min; to 27 to 35% B in 10 min; to 26% B in 0.1 min; hold at 26% B for 9.9 min; flow rate: 0.3 mL/min; pressure: 1029 psi (71 bar); column temp.: 50 °C; detector: Q-TOF (ESI-negative), XIC of the following m/z values: AMP-346, ADP-426, ATP-506, CMP-322, CDP-402, (CTP-482, GMP-362, GDP-442, GTP-522, UMP-323, UDP-403, UTP-483); injection: 10 μL; sample: 10-30 μg/mL of nucleotides (AMP, ADP, ATP, CMP, CDP, CTP, GMP, GDP, GTP, UMP, UDP, UTP) (80/20 acetonitrile/50 mM ammonium acetate in water, pH 5.3 with acetic acid)
• suitability: application for LC-MS
• application(s): pharmaceutical (small molecule)

Description

Analysis Note

Nucleotides are the building blocks of nucleic acids. They comprise a nitrogenous base, a sugar, and a phosphate group making them highly polar and ionic. This causes a challenge for typical reversed phase chromatography (RPC). These charged polar analytes are poorly retained by RPC, and thus typically chromatographed by either ion-exchange chromatography or RPC in conjunction with an ion-pairing reagent. Nucleotide analysis using ion-pair reagents or high concentration of phosphate buffers cause high background and ion source pollution for MS detection. HILIC separation is an alternative that permits sensitive MS detection and without the use of ion-pair reagents.

Legal Information

SeQuant is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Adenosine 5′-monophosphate sodium salt, from yeast, ≥99%
Adenosine 5′-diphosphate sodium salt, bacterial, ≥95% (HPLC)
Adenosine 5′-triphosphate disodium salt hydrate, BioXtra, ≥99% (HPLC), from microbial
Cytidine 5′-monophosphate, Sigma Grade, ≥99% (HPLC), synthetic, powder
Cytidine 5′-diphosphate sodium salt hydrate, from yeast, ≥95%
Cytidine 5′-triphosphate disodium salt, ~95%
Guanosine 5′-monophosphate disodium salt hydrate, from yeast, ≥99%
Guanosine 5′-diphosphate sodium salt, Type I, ≥96% (HPLC)
Guanosine 5′-triphosphate sodium salt hydrate, ≥95% (HPLC), powder
Uridine 5′-monophosphate, ≥98%
Uridine 5′-(trihydrogen diphosphate) sodium salt from Saccharomyces cerevisiae, 95-100%
Uridine 5′-triphosphate trisodium salt hydrate, from yeast, Type III, ≥96%
See All (12)