Determination of rimantadine in human urine by HPLC using a monolithic stationary phase and on-line post-column derivatization

Abstract

In the present study, we propose the first HPLC method coupled to postcolumn derivatization for the determination of rimantadine in human urine samples. The analyte and amantadine (internal standard) were isocratically separated using an RP monolithic stationary phase (100 × 4.6 mm id) with a mobile phase consisting of CH3OH/phosphate buffer (25 mmol/L, pH 3.0) at a volume ratio of 50:50. Postcolumn derivatization involved on-line reaction with o-phthalaldehyde (20 mmol/L) and N-acetyl-cysteine (5 mmol/L) at alkaline medium (100 mmol/L borate pH 11.0). Spectrofluorimetric detection at λ(ex)/λ(em) = 340/455 nm enabled the selective and sensitive determination of rimantadine in urine samples at a range of 50-500 ng/mL with an LOD of 5 ng/mL. Human urine samples were analyzed successfully after SPE using hydrophilic-lipophilic balanced RP cartridges (30 mg/mL, Oasis HLB). Recoveries ranged between 89.7 and 102.7%.

Trace analysis of rimantadine in human urine after dispersive liquid liquid microextraction followed by liquid chromatography–post column derivatization

“…In our case, an acidic mobile phase consisted of phosphate buffer and methanol was used for the separation on the analyte and ISTD on a reversed phase silica-based monolithic column [6]. Generally when such compounds are chromatographed at reversed phase conditions, secondary interactions occur between the positively charged amino group and the negatively charged free silanolic groups of the stationary phase.…”

“…Apart from mass spectrometricbased methods [12,25], derivatization is a common practice that has been widely used in the literature for the quantitation on RIM [6,8,10]. biological fluids) is rather challenging due to the lack of chromophore or fluorophore group in its chemical structure.…”

“…Moreover, a preconcentration step is typically required in order to facilitate the analysis at trace levels. Several sample clean-up techniques have been proposed for the determination of RIM in biological samples and animal-derived feed products including SPE [6,7], conventional liquid-liquid extraction [8][9][10][11], protein precipitation [12], QuEChERS [13], and carbon nanotubes-based dispersive SPE [14]. Supporting Information Table 1S contains the main analytical figures of merit of the above-mentioned methods including the analytical/sample preparation technique and the selected applications.…”

"Development and Validation of a Rp-HPLC Method for the Analysis of Rimantadine Hydrochloride in Medicinal Form"

“…New separation method for tricyclic antiviral drugs was developed 4 . Previous works included determination of RMT using IR Analysis 5 , online post column DRT 6 , fluorescent probe 7 , UV-visible spectrophotometry 8 , UHPLC coupled with orbitrap mass spectrometry 9 , use of AQSC for RMT analysis 10 , and simultaneous analysis using tandem mass spectrometry 11 . The present work focused on RP-HPLC method development 12 and its validation 13 .…”

Determination of Sudan dyes in chili pepper powder by online solid‐phase extraction with a butyl methacrylate monolithic column coupled to liquid chromatography with tandem mass spectrometry

“…Recently, polymer‐based monoliths as SPE sorbents have received considerable attention for the extraction and enrichment of analytes in matrices for environmental, food, and biological analyses because of their attractive merits . Compared with conventional particle‐packed SPE devices, polymer‐based monolithic columns can be easily fabricated within the device by in situ polymerization and possess the advantages such as availability of various monomers, versatility, and flexibility to modulate the surface chemistry.…”