"Center punch" and "whole spot" bioanalysis of apixaban in human dried blood spot samples by UHPLC-MS/MS.
Apixaban (Eliquis™) was developed by Bristol-Myers Squibb (BMS) and Pfizer to use as an antithrombotic/anticoagulant agent and has been recently approved for the prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation. A clinical study of apixaban, sponsored by BMS and Pfizer, included a pilot exploratory portion to evaluate the potential for future drug concentration monitoring using dried blood spot (DBS) sample collection. For DBS sample collection, a fixed blood volume was dispensed onto a DBS card by either regular volumetric pipette (venous blood collection) or capillary dispenser (finger prick blood collection). A 96-well semi-automated liquid-liquid extraction sample preparation procedure was developed to provide clean extracts for UHPLC-MS/MS quantitation. Assays using both partial-spot center punch and whole spot punch were developed and validated. The linear dynamic ranges for all the analyses were from 0.5 to 500 ng/mL. The coefficient of determination (r(2)) values was >0.9944 for all the validation runs. For the center punch approach, the intra-assay precision (%CV) was within 4.4% and inter-assay precision was within 2.6%. The assay accuracy, expressed as %Dev., was within ± 5.4% of the nominal concentrations. One accuracy and precision run was performed using the whole spot approach, the intra-assay precision (%CV) was within 7.1% and the accuracy was within ± 8.0% of the nominal concentrations. In contrast to the center punch approach, the whole spot approach eliminated the effect of hematocrit and high lipids on the analysis of apixaban in human DBS when an accurate sample blood volume was collected on DBS cards.
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Naiyu
N
Zheng
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States. Electronic address: long.yuan@bms.com.
Long
L
Yuan
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States. Electronic address: qin.ji@bms.com.
Qin C
QC
Ji
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Heidi
H
Mangus
Exploratory Clinical & Translational Research, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Yan
Y
Song
Exploratory Clinical & Translational Research, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Charles
C
Frost
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Jianing
J
Zeng
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Anne-Françoise
AF
Aubry
Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, United States.
Mark E
ME
Arnold
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D059625:Liquid-Liquid Extraction; D011720:Pyrazoles / Q000097:blood* / Q000737:chemistry; D011728:Pyridones / Q000097:blood* / Q000737:chemistry; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D011720:Pyrazoles; D011728:Pyridones; C522181:apixaban
Dried blood spot; Hematocrit effect; Liquid–liquid extraction; UHPLC–MS/MS; Whole spot punch