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

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

Materials

analytical column

Product No.
Description
Pricing

Discovery® HS C18 HPLC Column

5 μm particle size, L × I.D. 5 cm × 4.6 mm

SPE tube or plate

Product No.
Description
Pricing

Discovery® DSC-CN SPE Tube

bed wt. 500 mg, volume 3 mL, pk of 54

CONDITIONS

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

Description

Application

Sample Pre-treatment:
Dissove 1 g 1% hydrocortisone topical hand cream in 10 mL ethyl acetate. Dilute 2 mL of ethyl acetate-cream sample with 8 mL hexane:ethyl acetate (2:1, v/v).

Legal Information

Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany