Sensitive and rapid liquid chromatography-tandem mass spectrometry method for the determination of stavudine in human plasma |
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Authors: | Wiesner J L Sutherland F C W Smit M J van Essen G H Hundt H K L Swart K J Hundt A F |
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Institution: | FARMOVS-PAREXEL Clinical Research Organisation, Private Bag X09, Brandhof, Bloemfontein 9324, South Africa. lubbe.wiesner@farmovs-parexel.com |
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Abstract: | A sensitive method for the determination of stavudine in plasma was developed, using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The samples were extracted from plasma with Waters, Sep-Pak Vac, 100 mg, tC(18) solid-phase extraction (SPE) columns. Chromatography was performed on a Supelco Discovery C(18), 5 microm, 150 x 2 mm column with a mobile phase consisting of ammonium acetate (0.01 M)-acetonitrile-methanol (800:100:100, v/v/v) at a flow-rate of 0.3 ml/min. Detection was achieved by an Applied Biosystems API 2000 mass spectrometer (LC-MS-MS) set at unit resolution in the multiple reaction monitoring mode (MRM). Atmospheric pressure chemical ionization (APCI) was used for ion production. The mean recovery for stavudine was 94% with a lower limit of quantification set at 4 ng/ml. This assay method makes use of the increased sensitivity and selectivity of mass spectrometric (MS-MS) detection to allow for a more rapid (extraction and chromatography) and selective method for the determination of stavudine in human plasma than has previously been described. |
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