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Title
Contribution to the development and validation of a high performance liquid chromatography by the UV detection method for isoniazid and omeprazole determination
Authors
MAGDA COSTIN BOGDAN CIOROIU LUMINITA AGOROAEI and ELENA BUTNARU
Received
August 15, 2011
Published
Volume 46 Issue 7-8 September-October
Keywords
HPLC-UV, validation, isoniazid, omeprazole
Abstract
The objective of this study was to develop and validate a method for isoniazid and omeprazole determination in human
serum. Solid phase extraction techniques have been used for sample preparation. The analytical method was applied on
a Thermo Fisher Scientific Surveyor Plus chromatographic system, equipped with an autosampler and UV-VIS with
diode array detector. Separation was performed on a C8 chromatographic column Octasilil (Purospher RP8) of the 250
mm x 4.6 mm i.d. type (5 μm). A mixture of 10 mM triethylamine (with a pH value of 10.5): acetonitrile (67:33, v/v)
has been used as mobile phase. The obtained retention times were the following: 2.323 min for isoniazid; 3.497 min for
2-pyridylamine (used as an internal standard); 4.013 min for omeprazole and 6.837 min, respectively, for lansoprazole
(used as internal standard). Detection was in UV at 260 nm for isoniazid and 2-pyridylamine, and at 300 nm,
respectively, for omeprazole and lansoprazole. The method is linear, selective, accurate and precise in the 50-5000
ng/mL concentration range.
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