Sx. Peng et al., Automated 96-well SPE and LC-MS-MS for determination of protease inhibitors in plasma and cartilage tissues, ANALYT CHEM, 72(8), 2000, pp. 1913-1917
Bioanalytical methods based on automated solid-phase extraction (SPE) and h
igh-performance liquid chromatography with electrospray tandem mass spectro
metry (LC-MS-MS) have been developed and utilized for the determination of
MMP inhibitors in plasma and cartilage tissues. The SPE methods were automa
ted using a 96-wed extraction plate and a 96-channel programmable liquid-ha
ndling workstation. The LC-MS-MS methods were developed using a rapid gradi
ent LC separation, followed by sample introduction through an ionspray inte
rface in the positive ion mode and tandem mass spectrometric detection with
selected reaction monitoring. In the optimized SPE methods, crude plasma o
r ground cartilage supernatant samples were loaded onto an SPE plate to rem
ove proteins and other interfering components in the matrixes to render rel
atively dean extracts for LC-MS-MS analysis. Compared to the simple plasma
protein precipitation method, the automated SPE method afforded significant
time-saving in sample preparation and improved sensitivity in MS detection
, The methods were validated and successfully applied to the analysis of pr
otease inhibitors in plasma and cartilage tissues.