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HomeSmall Molecule HPLCLC/MS Analysis of Nucleotides SeQuant® ZIC®-cHILIC – Effect of pH

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

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

Figure 1.

Conditions

InstrumentAgilent® 6530 Q-TOF LC/MS
ColumnSeQuant® 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)
Gradient26 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 Rate0.3 mL/min
Pressure986 psi
Column Temp.40 °C
DetectorQ-TOF (ESI-negative)
Detector OtherXIC 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
Injection10 µL

Description

Analysis NoteZIC®-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 IndustryLife Science and Biopharma 
Legal InformationSeQuant is a trademark of Merck KGaA, Darmstadt, Germany
LiChrosolv is a trademark of Merck KGaA, Darmstadt, Germany
LiChropur is a trademark of Merck KGaA, Darmstadt, Germany
Application No.ASC140218I
Materials
产品编号产品名称说明价格
MSQC16SILu™Lite SigmaMAb Adalimumab Monoclonal Antibodyrecombinant, expressed in CHO cells
31417葡聚糖 来源于肠系膜明串珠菌analytical standard, Mw 5,000
I4506IgG 来源于人类血清reagent grade, ≥95% (SDS-PAGE), essentially salt-free, lyophilized powder
50933盐酸胍 盐酸盐BioUltra, ≥99.5% (AT)
09830碳酸氢铵BioUltra, ≥99.5% (T)
F8435糖苷酶F 来源于脑膜脓毒性伊丽莎白菌lyophilized powder, recombinant, expressed in E. coli
EMS0011Low-Artifact Digestion BufferMinimizes artifactual deamidation and oxidation during trypsin digestion prior to peptide mapping
MRCF0R030Microcon®离心超滤管NMWCO 30 kDa, Ultracel® regenerated cellulose membrane (low binding), sample volume 0.5 mL
156159氰基硼氢化钠reagent grade, 95%
PHR1252普鲁卡因胺 盐酸盐Pharmaceutical Secondary Standard; Certified Reference Material
D8418二甲基亚砜
695092乙酸glacial, ACS reagent, ≥99.7%
55465-UDiscovery® Glycan SPE Tubebed wt. 50 mg, volume 1 mL, pk of 108
1.00029乙腈hypergrade for LC-MS LiChrosolv®
VM20VM20真空歧管For processing up to 20 plasmid purification samples simultaneously
50994-UBIOshell Glycan HPLC ColumnsL × I.D. 15 cm × 2.1 mm, 2.7 μm particle size
1.00029乙腈hypergrade for LC-MS LiChrosolv®
70221甲酸铵eluent additive for LC-MS, LiChropur™, ≥99.0%
5.33002甲酸98％-100％for LC-MS LiChropur™
ZIQ7000T0CMilli-Q® IQ 7000 超纯水净化系统output: type 1 water (18.2 MΩ·cm), the most advanced Milli-Q® ultrapure (Type 1) water purification system that is intelligent, intuitive, and green.
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