Evaluation of high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA) and particle concentration fluorescence immunoassay (PCFIA) methods for the screening, quantitation and pharmacokinetic study of furosemide in horses.

The study involves a comparison of three methods – High Performance Liquid Chromatography (HPLC), Enzyme Linked Immunosorbent Assay (ELISA) and Particle Concentration Fluorescence Immunoassay (PCFIA), for their efficacy in detection, quantity measurement and pharmacokinetic studies of the drug Furosemide in horses. Results showed HPLC to be the most reliable, whereas ELISA and PCFIA were found to generate considerable data inconsistencies.

Research Methods Used

• The researchers used three different methods in the study – High Performance Liquid Chromatography (HPLC), Enzyme Linked Immunosorbent Assay (ELISA) and Particle Concentration Fluorescence Immunoassay (PCFIA).
• The samples for the analysis were plasma and urine obtained from thoroughbred mares that were intravenously dosed with Furosemide.

Findings of the Research

• The study found the HPLC method to be a time-intensive process, but it provided notable accuracy and repeatability in the quantitative analysis of Furosemide in equine plasma and urine samples.
• The ELISA and PCFIA methods were discovered to be quicker and simpler than HPLC, and had high sensitivity for Furosemide screening.
• However, ELISA and PCFIA methods projected significant variability in their results, raising questions on their reliability for quantitative analysis.
• When compared at high Furosemide concentrations, PCFIA displayed better quantitative results in comparison to ELISA.
• On the contrary, at low or trace Furosemide levels, PCFIA delivered false positive results, which were not approved or confirmed by ELISA or HPLC.

Comparison with Previous Research

• The HPLC method’s pharmacokinetic values matched well with already established values from prior research using gas chromatographic data, further establishing the validity and efficacy of HPLC.
• In contrast, the data acquired from ELISA and PCFIA methods were deemed unsuitable for pharmacokinetic calculations.

Implications for Future Research

• The study’s findings provide valuable insights for future inquiries regarding drug analysis methods in veterinary studies, specifically in the detection and quantification of Furosemide.
• The results underline the importance of using methods like HPLC that may be more time-consuming but provide accurate and reproducible results, as opposed to faster but inconsistent methods like ELISA and PCFIA.