Type

Journal Article

Authors

Robert O'Connor
Martin Clynes
Peter O'Gorman
Rajesh Rajpal
Kasper Pedersen
Justine Meiller
Louise Sewell
Sandra Roche

Subjects

Microbiology

Topics
chromatography liquid blood sensitivity and specificity culture media humans cell line tumor reproducibility of results drug stability chemistry cell extracts liquid liquid extraction thalidomide isolation purification

Development, validation and application of a sensitive LC-MS/MS method for the quantification of thalidomide in human serum, cells and cell culture medium. (2011)

Abstract A simple, robust, sensitive and selective liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the quantification of thalidomide was developed and validated. The method was applied to thalidomide quantification in three different types of biological samples. Thalidomide was extracted from human serum (100 μL), cells (2.5 × 10(5)), or cell culture media (100 μL) by LLE and separated on a Prodigy C18 (150 mm × 4.0 mm, 5 μm i.d.) column with isocratic elution using water/acetonitrile (70/30, v/v) 0.1% formic acid, at a flow rate of 0.5 mL/min, with umbelliferone (600 ng/mL) as an internal standard. Thalidomide was quantified using a triple quadrupole mass spectrometer operated in multi-reaction-monitoring mode using positive electrospray ionisation. The method was validated in two separate thalidomide concentration ranges; human serum (0.05-20 μg/mL) and in vitro cells (0.78-50 ng) with an inter-day precision of 1.8% and 1.9% and average accuracy of 100% and 101% in serum and cells respectively. Despite the use of small sample volume, the limit of quantification for thalidomide in serum was determined to be 3 ng/mL. The method was successfully employed to measure levels of thalidomide in cancer patient serum and cell culture model systems. Although cellular levels were quantifiable, thalidomide was shown to be unstable under in vitro conditions with a half life of approximately 2 h. In patient samples, circulating serum levels showed a broad correlation with dose and uncovered some patient compliance issues.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Robert O'Connor, Martin Clynes, Peter O'Gorman, Rajesh Rajpal, Kasper Pedersen, Justine Meiller, Louise Sewell, Sandra Roche

Experts in our system

1
Robert O'Connor
Dublin City University
Total Publications: 71
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Peter O'Gorman
Dublin City University
Total Publications: 15
 
4
Justine Meiller
Dublin City University
Total Publications: 7
 
5
Sandra Roche
Dublin City University
Total Publications: 6