A validated microfluidics‐based LC‐chip‐MS/MS method for the quantitation of fluoxetine and norfluoxetine in rat serum

Abstract

An increasing number of quantitative bioanalyses need to be performed on samples available in limited volumes, such as pharmacokinetic studies on small animals. In this context, microfluidic systems as the LC-chip device coupled to a mass spectrometer combine small sample volume requirements and high sensitivity. In this study, we present the development of a microfluidics-based method for fluoxetine (FLX) and norfluoxetine (NFL) quantitation dedicated to pharmacokinetic investigations in the rat serum. Using the methodology of experimental design, LC parameters were optimised in terms of peak resolution, analysis time, and sensitivity. An SPE method was then developed for serum samples on miniaturised 96-well plates containing a mixed-mode strong cation exchanger that provided very clean extracts with good analyte recovery (≥66.0%). The total SPE-LC-MS/MS process required only 20 μL per sample and the method provided a good sensitivity in a total run time of 12 min. Finally, the developed method for FLX and NFL quantitation in rat serum was fully validated. After having selected the most appropriate regression model on the basis of the accuracy profiles, method selectivity, trueness, precision, accuracy and linearity were demonstrated.

Quantitative profiling of endocannabinoids and related N-acylethanolamines in human CSF using nano LC-MS/MS

“…In addition, the reduced flow rate of nano LC compared with conventional LC can improve sensitivity through an increase in the ionization efficiency (29). Using microfluidic chip-based technology, incorporating an enrichment column, a six-port valve, a separation column, and a nano electrospray tip allows the coupling of miniaturized separation with a nano ESI source (30). This design results in lower dead volumes and fewer connections compared with classical nano LC equipment, thereby leading to increased sensitivity and robustness by less peak broadening and dilution effects during transfer of the sample to the LC column and later to the LC-MS interface (31).…”

Recent Developments in Microfluidic Chip-Based Separation Devices Coupled to MS for Bioanalysis

“…In addition to analyzing biological macromolecules, microfluidic chip-based CE-MS and LC-MS systems have also been utilized for determining small molecules, such as pharmaceutical ingredients, drugs of abuse and drug metabolites, as well as tumor/disease-associated biomarkers for drug discovery/development and clinical applications [107][108][109][110][111][112]. A recent review discussed the developments of several chip-based LC-MS systems for the determination of small molecules in bioanalytical applications [8].…”

“…The proposed chip-based LC-MS was also successfully applied for profiling of six varieties of soybean samples. Houbart et al described a chip-based LC-MS/MS method for quantifying pharmaceutical drugs and drug metabolites in rat serum [111]. Thawed rat serum samples were vortexed, centrifuged, and subjected to SPE.…”

“…Reverse-phase LC Peptides/proteins Protein digests, cell lysates [65,66,68,81,83,84,95] Serum, plasma, cerebrospinal fluid, milk, nail [74,75,89,96,97] Pharmaceutical drugs; drug metabolites Serum, urine, hair [78,79,111,112] Biomarkers Urine, nail [80,107] Bioactive compounds Grapes, soybeans [90,110] HILIC Oligosaccharides Glycan digests [76,103] Graphitized carbon Oligosaccharides Milk, serum, protein digests [77,101,102,[104][105][106] LC-CE Peptides/proteins Cell lysate [100] Executive summary…”

Simultaneous Determination of Methadone, Fluoxetine, Venlafaxine and Their Metabolites in Rat Plasma by UPLC–MS/MS for Drug Interaction Study

“…study the drug-drug interaction of methadone with antidepressive drugs. Though various methods were reported in the literature for estimation of methadone and fluoxetine/venlafaxine separately or in combination with other drugs [12][13][14][15][16], no method had been reported for simultaneous determination of these drugs using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). In the present study, we developed a UPLC-MS/MS method for the simultaneous determination of methadone, fluoxetine, venlafaxine and their main metabolites using diazepam as an internal standard in rat plasma, which was fully validated in terms of specificity, linearity, matrix effect and recovery, accuracy, precision and stability.…”