Development and validation of a semi-automated assay for the highly sensitive quantification of Biolimus A9 in human whole blood using high-performance liquid chromatography-tandem mass spectrometry.
Drug-eluting stents are sustained-release intra-coronary devices that are usually coated with a few hundred micrograms of drug. Measuring the drugs that are released over weeks in order to assess human pharmacokinetics is a challenge that requires assays with high sensitivity. We developed and validated a semi-automated LC-MS/MS assay for the quantification of Biolimus A9, a proliferation signal inhibitor that was specifically developed for coating on drug-eluting stents in human EDTA blood. The only manual step was the addition of a zinc sulfate/methanol protein precipitation solution which included the internal standard. Samples were injected into the HPLC and extracted online. The assay had the following performance characteristics: range of reliable response 0.01-100 ng/mL (r(2)>0.99), inter-day accuracy (0.033 ng/mL): 111.7%, and inter-day precision: 8.6%. There was no ion suppression, matrix interferences or carry-over. Extracted samples were stable in the autosampler at +4 degrees C for at least 24 h and could undergo three freeze-thaw cycles. The assay, with a lower limit of detection of 333 fg/mL and a lower limit of quantitation of 10 pg/mL, was sufficiently sensitive and robust for quantifying Biolimus A9 in clinical trials after i.v. injection and after stent implantation.
Clavijo
Claudia
C
Clinical Research & Development, Department of Anesthesiology, University of Colorado Denver, Aurora, CO 80045-7503, USA.
Strom
Tobin
T
Moll
Vanessa
V
Betts
Ronald
R
Zhang
Yan Ling
YL
Christians
Uwe
U
Bendrick-Peart
Jamie
J
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D054855:Drug-Eluting Stents; D006801:Humans; D015203:Reproducibility of Results; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*
C518022:umirolimus; D020123:Sirolimus