HPLC Analysis of Hormone on Chromolith^® Performance 100-3mm

column

Chromolith Performance, 100 x 3 mm (1.52001.0001)

mobile phase

[A] acetonitrile; [B] 90% Phosphatbuffer 10mM pH=7.0 + 10% Acetonitrile;

gradient

Initial composition: 0-0.5min 45%A 212μl/min, 0.5-0.6min 45%A to 35%A 212μl/min, 0.6-3min 35%A 212μl/min to 637μl/min, 3.0-3.1min 35%A to 55%A 637μl/min, 3.1-8min 55%A 637μl/min

pressure

653 psi 9-45 Bar

column temp.

30 °C

detector

220 nm cell: 11 μL

injection

1 μL

sample

IPrednisolone 0,6mg/ml, beta-Estradiol 0,4mg/ml, 17-alpha-Ethinyl-estradiol 0,4mg/ml, Estrone 0,5mg/ml, Progesterone 0,8mg/ml solved in Acetonitrile