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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 as indicated with acetic acid in water (50 mM ammonium acetate, pH 6.93 in water not adjusted)
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	986 psi
Column Temp.	40 °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

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Analysis Note	ZIC^®-cHILIC is a densely bonded zwitterionic stationary phase with phosphorylcholine functional groups covalently attached to silica. The stationary phase has an overall 1:1 charge balance between the permanent negative phosphate groups close to the silica support and the distal positively quaternary ammonium groups. It has though a slight positive surface charge because of the spatial arrangement. A true zwitterionic stationary phase, with permanent strong acidic and basic functionalities, provide hydrophilic partitioning combined with weak ionic interactions and makes the phase virtually pH independent as shown in this analysis.
Featured Industry	Life Science and Biopharma
Legal Information	SeQuant is a trademark of Merck KGaA, Darmstadt, Germany \r\nLiChrosolv is a trademark of Merck KGaA, Darmstadt, Germany \r\nLiChropur is a trademark of Merck KGaA, Darmstadt, Germany
Application No.	ASC140218I

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LC/MS Analysis of Nucleotides SeQuant® ZIC®-cHILIC – Effect of pH

Materials

LC/MS Analysis of Nucleotides SeQuant^® ZIC^®-cHILIC – Effect of pH