Automated liquid–liquid extraction based on 96-well plate format in conjunction with ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) for the quantitation of methoxsalen in human plasma

A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid–liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) has been developed for the determination of methoxsalen in human plasma. Plasm...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2008-09, Vol.872 (1), p.167-171
Main Authors: Yadav, Manish, Contractor, Pritesh, Upadhyay, Vivek, Gupta, Ajay, Guttikar, Swati, Singhal, Puran, Goswami, Sailendra, Shrivastav, Pranav S.
Format: Article
Language:English
Subjects:
96-Well plate liquid–liquid extraction
Analysis
Analytical, structural and metabolic biochemistry
Automation
Biological and medical sciences
Chromatography, Liquid - methods
Fundamental and applied biological sciences. Psychology
General pharmacology
High throughput
Human plasma
Humans
Medical sciences
Methoxsalen
Methoxsalen - blood
Pharmacology. Drug treatments
Photosensitizing Agents - blood
Reference Standards
Reproducibility of Results
Sensitivity and Specificity
Tandem Mass Spectrometry - methods
UPLC–MS/MS
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Summary:A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid–liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) has been developed for the determination of methoxsalen in human plasma. Plasma samples with ketoconazole as internal standard (IS) were prepared by employing 0.2 mL human plasma in ethyl acetate:dichloromethane (80:20, v/v). The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column using isocratic mobile phase, consisting of 10 mM ammonium formate and acetonitrile (60:40, v/v), at a flow rate of 0.5 mL/min. The linear dynamic range was established over the concentration range 1.1–213.1 ng/mL for methoxsalen. The method was rugged and rapid with a total run time of 1.5 min. It was successfully applied to a pivotal bioequivalence study in 12 healthy human subjects after oral administration of 10 mg extended release methoxsalen formulation under fasting condition.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2008.06.058