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. 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.
Yadav
Manish
M
Bioanalytical Research Department, Veeda Clinical Research, Ambawadi, Ahmedabad 3800015, India.
Contractor
Pritesh
P
Upadhyay
Vivek
V
Gupta
Ajay
A
Guttikar
Swati
S
Singhal
Puran
P
Goswami
Sailendra
S
Shrivastav
Pranav S
PS
D001331:Automation; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D008730:Methoxsalen / Q000097:blood*; D017319:Photosensitizing Agents / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D017319:Photosensitizing Agents; D008730:Methoxsalen