Development, validation, and implementation of an UHPLC-MS/MS method for the quantitation of furosemide in infant urine samples.

TitleDevelopment, validation, and implementation of an UHPLC-MS/MS method for the quantitation of furosemide in infant urine samples.
Publication TypeJournal Article
Year of Publication2022
AuthorsVedar C, Bamat NA, Zuppa AF, Reilly ME, Moorthy GS
JournalBiomed Chromatogr
Volume36
Issue3
Paginatione5262
Date Published2022 Mar
ISSN1099-0801
KeywordsChild, Chromatography, High Pressure Liquid, Furosemide, Humans, Infant, Newborn, Infant, Premature, Reproducibility of Results, Tandem Mass Spectrometry
Abstract

Furosemide is a diuretic drug used to increase urine flow in order to reduce the amount of salt and water in the body. It is commonly utilized to treat preterm infants with chronic lung disease of prematurity. There is a need for a simple and reliable quantitation of furosemide in human urine. We have developed and validated an ultra-high performance liquid chromatography-tandem mass spectrometry method for furosemide quantitation in human urine with an assay range of 0.100-50.0 μg/ml. Sample preparation involved solid-phase extraction with 10 μl of urine. Intra-day accuracies and precisions for the quality control samples were 94.5-106 and 1.86-10.2%, respectively, while inter-day accuracies and precision were 99.2-102 and 3.38-7.41%, respectively. Recovery for furosemide had an average of 23.8%, with an average matrix effect of 101%. Furosemide was stable in human urine under the assay conditions. Stability for furosemide was shown at 1 week (room temperature, 4, -20 and -78°C), 6 months (-78°C), and through three freeze-thaw cycles. This robust assay demonstrates accurate and precise quantitation of furosemide in a small volume (10 μl) of human urine. It is currently being implemented in an ongoing pediatric clinical study.

DOI10.1002/bmc.5262
Alternate JournalBiomed Chromatogr
PubMed ID34648199
PubMed Central IDPMC8881385
Grant ListK23 HD101651 / HD / NICHD NIH HHS / United States
K23HD101651 / NH / NIH HHS / United States