VALIDATION OF HPLC- UV METHOD FOR ANALYSIS OF OMEPRAZOLE IN PRESENCE OF ITS METABOLITES IN HUMAN PLASMA.
CRISTINA IUGA, MIRELA MOLDOVAN, ADINA POPA,
S.E. LEUCUŢA
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Abstract:

We present a simple and validated HPLC method for measuring omeprazole, in presence of its two main metabolites, in plasma. Omeprazole, hydroxyomeprazole and omeprazole sulfone were extracted from plasma samples with a mixture of dichloromethane–diethyl ether. HPLC separation was achieved using a Kromasil C₁₈ 150mm x 4.6 mm i.d (5µm) column. The mobile phase was CH₃OH:H₂O (55:45 v/v) containing triethylamine (TEA) 1% at pH=7 with H₃PO₄ 85%. Retention times were 8.0 min for omeprazole, 3.9 min for hydroxy-omeprazole, 7.0 min for omeprazole sulfone and 4.2 min for internal standard (phenacetine). Detection (UV at 302 nm) of analytes was linear in a range from 25 to 500ng/ml. This method is stable, reproducible and has practical advantages such as low cost.

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