LC/MS Analysis of Nucleotides on SeQuant ZIC-cHILIC
Conditions | |
|---|---|
| instrument | Agilent® 6530 Q-TOF LC/MS |
| column | SeQuant ZIC-cHILIC, 10 cm x 2.1 mm, 3.0 µm |
| 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 | 1059 psi |
| column temp. | 50 °C |
| detector | Q-TOF (ESI-negative) |
| detector other | 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 |
Description | |
|---|---|
| Analysis Note | HILIC separation is an alternative that permits sensitive MS detection and without the use of ion-pair reagents. 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 and is undesirable compared to HILIC. |
| Featured Industry | Life Science and Biopharma |
| Legal Information | SeQuant is a trademark of Merck KGaA, Darmstadt, Germany LiChrosolv is a trademark of Merck KGaA, Darmstadt, Germany |
| Application No. | ASC140218E |
Materials
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