Investigation of EDTA anticoagulant in plasma to improve the throughput of liquid chromatography/tandem mass spectrometric assays.
In this study, EDTA and heparin are compared as anticoagulants with respect to their efficiency in preventing clot formation in plasma samples that were subsequently analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). A pilot in vivo pharmacokinetic study for the drug chlorpheniramine was conducted in which both EDTA and heparin plasma samples were collected simultaneously. All conditions except the anticoagulant were held constant during the pharmacokinetic study. Bioanalytical results were compared from samples transferred by manual pipette and by an automated liquid handler workstation. The concentration of chlorpheniramine in samples was determined by LC/MS/MS. Results from the analysis of variances (ANOVA) of log-transformed plasma chlorpheniramine concentrations were used to calculate 90% confidence intervals for the ratio least-squares mean values for anticoagulants and for transfer methods. Analytical concentrations of the drug chlorpheniramine were equivalent in heparin- and EDTA-containing plasma. Results suggest that the failure rate for transfer of EDTA plasma (50 micro L by automated workstation or manually) is less than that for heparinized plasma. As a consequence of these results, the vast majority of plasma samples in our laboratories are now collected in EDTA, which allows for use of automated sample transfer resulting in a three-fold timesaving over manual transfer using a single-channel pipette. The ability to use automation has resulted in improved efficiency and cost savings.
Department of Pharmacokinetics Dynamics and Metabolism, Pfizer Global R&D, Ann Arbor Laboratories, 2800 Plymouth Road, Ann Arbor, MI 48105, USA. nalini.sadagopan@pfizer.com
Nalini P
NP
Sadagopan
Wenlin
W
Li
Jack A
JA
Cook
Betsy
B
Galvan
David L
DL
Weller
Scott T
ST
Fountain
Lucinda H
LH
Cohen
D000818:Animals; D000925:Anticoagulants / Q000737:chemistry*; D002744:Chlorpheniramine / Q000097:blood / Q000493:pharmacokinetics; D002853:Chromatography, Liquid; D004492:Edetic Acid / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry; D006634:Histamine H1 Antagonists / Q000097:blood / Q000493:pharmacokinetics; D013058:Mass Spectrometry; D010949:Plasma / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results
D000925:Anticoagulants; D006634:Histamine H1 Antagonists; D002744:Chlorpheniramine; D006493:Heparin; D004492:Edetic Acid