HPLC Analysis of Hydrocortisone in Topical Cream on Discovery^® HS C18 after SPE using Discovery^® DSC-Si

sample preparation

SPE (Solid Phase Extraction)

sample addition

hydrocortisone topical hand cream in hexane:ethylacetate

SPE tube/cartridge

Discovery DSC-Si, 500 mg/3 mL (52695-U)

condition

1 mL hexane:ethyl acetate (2:1, v/v)

sample addition

1 mL

washing

1 mL hexane:ethyl acetate (2:1, v/v)

elution

1 mL methanol:DI water (1:1, v/v)

eluate post-treatment

dilute eluate to 2 mL with methanol:DI water (1:1, v/v)

column

Discovery HS C18, 15 cm x 4.6 mm I.D., 5 μm particles (568520-U)

mobile phase

[A] methanol; [B] DI water (1:1, A:B)

flow rate

1 mL/min

column temp.

35 °C

detector

UV, 220 nm

injection

10 μL