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- Posts: 22
- Joined: Thu Apr 25, 2013 9:21 am
I could use some help here
I´m trying to measure creatinine by HPLC.
This is my setup:
Column:
Zorbax SCX 300, strong cation exchange, (Agilent), 50 mm x 2.1 mm, 5μ particle size with in-front five micron SCX guard column.
Solvent:
5mM sodium acetate
adjust to pH 4.1 with glacial acetic acid
degassed with 0.2micron filter.
creatinine anhydrous >98% (Sigma aldrich) disolved in solvent to give a 256 mM stock solution.
Temperature and flow: Column run at 50°C. at a flow of 1 ml/min. Backpressure at around 2200 - 2300 psi. Run time was 10 min. These parameters are set 45 – 60 min before run start to ensure a solid baseline (equilibration).
Detection: UV (deuterium source) at 225 nm (peptides).
Injection volume: 10 μl from autosampler. All samples are run in duplicates. Temperature of autosampler tray was 18 ± 0.5°C.
I keep getting tailing peaks during run...
Can anyone help with this??