Sensitive and rapid liquid chromatography-tandem mass spectrometry method for the determination of stavudine in human plasma.
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.
Wiesner
J L
JL
FARMOVS-PAREXEL Clinical Research Organisation, Private Bag X09, Brandhof, Bloemfontein 9324, South Africa. lubbe.wiesner@farmovs-parexel.com
Sutherland
F C W
FC
Smit
M J
MJ
van Essen
G H
GH
Hundt
H K L
HK
Swart
K J
KJ
Hundt
A F
AF
D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D018894:Reverse Transcriptase Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D012680:Sensitivity and Specificity; D018119:Stavudine / Q000097:blood* / Q000493:pharmacokinetics
D018894:Reverse Transcriptase Inhibitors; D018119:Stavudine