Demonstration of direct bioanalysis of drugs in plasma using nanoelectrospray infusion from a silicon chip coupled with tandem mass spectrometry.
Quantitative bioanalysis by direct nanoelectrospray infusion coupled to tandem mass spectrometry has been achieved using an automated liquid sampler integrated with an array of microfabricated electrospray nozzles allowing rapid, serial sample introduction (1 min/ sample). Standard curves prepared in human plasma for verapamil (r2 = 0.999) and its metabolite norverapamil (r2 = 0.998) were linear over a range of 2.5-500 ng/ mL. Based on the observed precision and accuracy, a lower limit of quantitation of 5 ng/mL was assigned for both analytes. Sample preparation consisted of protein precipitation with an organic solvent containing the structural analogue gallopamil as an internal standard. Protein precipitation was selected both to maximize throughput and to test the robustness of direct nanoelectrospray infusion. Aliquots of supernatant (10 pL) were transferred to the back plane of the chip using disposable, conductive pipet tips for direct infusion at a flow rate of 300 nL/min. Electrospray ionization occurred from the etched nozzles (30-microm o.d.) on the front of the chip, initiated by a voltage applied to the liquid through the pipet tip. The chip was positioned near the API sampling orifice of a triple quadrupole mass spectrometer, which was operated in selected reaction monitoring mode. Results are presented that document the complete elimination of system carry-over, attributed to lack of a redundant fluid path. This technology offers potential advantages for MS-based screening applications in drug discovery by reducing the time for methods development and sample analysis.
Dethy
Jean-Marie
JM
Lilly Development Center S.A., Eli Lilly and Company, Mont-Saint-Guibert, B- 1348, Belgium. JMDETHY@Lilly.com
Ackermann
Bradley L
BL
Delatour
Claude
C
Henion
Jack D
JD
Schultz
Gary A
GA
D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D004364:Pharmaceutical Preparations / Q000097:blood; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization; D014700:Verapamil / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism
D004364:Pharmaceutical Preparations; C016904:norverapamil; D014700:Verapamil