Development, validation, and application of a simple UPLC-MS/MS method for simultaneous quantification of five...

Area of Science:

• Analytical Chemistry
• Pharmacology
• Clinical Chemistry

Background:

• Traditional antipsychotics (APs) are crucial for managing psychosis.
• Accurate quantification of APs in human plasma is essential for therapeutic drug monitoring.
• Existing methods may lack the sensitivity or efficiency required for routine clinical use.

Purpose of the Study:

• To develop and validate a sensitive UPLC-MS/MS method for simultaneous quantification of five traditional antipsychotics.
• To establish a reliable analytical technique for measuring chlorprothixene, perphenazine, fluphenazine, thioridazine, and promethazine in human plasma.

Main Methods:

• Ultra-Performance Liquid Chromatography coupled with tandem Mass Spectrometry (UPLC-MS/MS).
• Simple liquid-liquid extraction for sample preparation.
• Gradient elution using a C18 column with a mobile phase of formic acid in water and acetonitrile.
• Carbamazepine used as an internal standard.

Main Results:

• The method demonstrated excellent linearity (r > 0.995) over the range of 2-250 ng/mL for all five antipsychotics.
• Low limits of quantification (2 ng/mL) and detection (0.1-0.5 ng/mL) were achieved.
• High precision (RSD < 10%) and accuracy (relative error -5.70 to 7.20%) were confirmed, with recoveries between 70-109%.

Conclusions:

• The developed UPLC-MS/MS method is simple, economical, sensitive, and accurate.
• This validated method is suitable for the simultaneous quantification of five traditional antipsychotics in human plasma.
• The method supports effective therapeutic drug monitoring and clinical management of patients on these medications.