Highly sensitive UPLC-MS/MS method for the quantification of paromomycin in human plasma

Area of Science:

• Analytical Chemistry
• Pharmacology
• Clinical Diagnostics

Background:

• Leishmaniasis treatment requires accurate drug monitoring.
• Paromomycin is a key antileishmanial agent.
• Quantifying paromomycin in human plasma presents analytical challenges.

Purpose of the Study:

• To develop and validate a highly sensitive analytical method for quantifying paromomycin in human plasma.
• To establish a reliable assay for supporting pharmacokinetic studies of paromomycin.

Main Methods:

• Ultra-high performance liquid chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS).
• Isocratic ion-pair chromatography utilizing heptafluorobutyric acid.
• Simple protein precipitation for sample preparation using trichloroacetic acid.

Main Results:

• The method achieved a low limit of quantification (5 ng/mL) with excellent linearity (r²≥0.997).
• High accuracy and precision were demonstrated, meeting international criteria.
• Paromomycin stability was confirmed under various storage conditions, with specific handling requirements noted (acidic conditions, polypropylene tubes).

Conclusions:

• A validated, sensitive, and robust UPLC-MS/MS assay for paromomycin in human plasma was successfully developed.
• The assay is suitable for application in pharmacokinetic studies, particularly in visceral leishmaniasis patients.
• The method addresses challenges related to paromomycin's adsorption to glass surfaces.