Sensitive and rapid method for the determination of thalidomide in human plasma and semen using solid-phase extraction and liquid chromatography-tandem mass spectrometry.
Liquid chromatography-tandem mass spectrometric assays were developed for the sensitive, rapid and high throughput bioanalyses of thalidomide in human plasma and semen. The matrices were first stabilized with 0.025 M Sorensen's citrate buffer at pH 1.5 to prevent spontaneous hydrolysis. Buffered thalidomide was stable when stored at room temperature for 24 h and for up to three freeze-thaw cycles. Samples were extracted using SPE cartridges. Extracts were then injected into the LC-MS-MS equipped with a reversed-phase column and an APCI interface in the negative ion mode. Calibration curves for both matrices were linear with r>0.99 from 2 to 250 ng/ml and ng/g. Inter-assay precision (RSD) of plasma and semen calibration standards were 2.6-11.6 and 1.9-12.4%, respectively. Recoveries from plasma and semen were greater than 69 and 78%, respectively. Batch sizes of 100 samples per matrix were analyzed with a total run time of 5 h. The methods successfully determined concentrations of thalidomide from a clinical study to levels as low as 7 ng/ml plasma and 8 ng/g semen, respectively.
Teo
Steve K
SK
Celgene Corporation, Warren, NJ 07059, USA. steo@celgene.com
Chandula
Reddy S
RS
Harden
Jill L
JL
Stirling
David I
DI
Thomas
Steve D
SD
D000328:Adult; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004311:Double-Blind Method; D006679:HIV Seropositivity / Q000378:metabolism*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D010919:Placebos; D015203:Reproducibility of Results; D012661:Semen / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013792:Thalidomide / Q000032:analysis* / Q000097:blood
D010919:Placebos; D013792:Thalidomide