HPLC Analysis of Hormone on Chromolith^® High Resolution RP-18e

column

Chromolith High Resolution RP-18 endcapped, 100 x 4.6 mm (1.52022.0001)

mobile phase

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

gradient

Initial composition: 0-0.5min 45%A 0.5ml/min; 0.5-0.6min 45%A to 35%A 0.5ml/min; 0.6-3min 35%A 0.5ml/min to 1.5ml/min; 3.0-3.1min 35%A to 55%A 1.5ml/min; 3.1-8min 55%A 1.5ml/min

column temp.

30 °C

detector

220 nm (cell: 11 μL)

injection

1 μL

sample

Prednisolone 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