Abstract
A simple method for quantification of citalopram in mice plasma and hair was developed and validated using liquid chromatography tandem mass spectrometry (LC–MS/MS). The procedure involves a protein precipitation extraction of citalopram and desipramine (internal standard) with methanol from mice plasma. On the other hand, hair samples were incubated overnight with methanol at 45°C followed by μ-SPE (OMIX Tip). The analysis was performed by resolving analytes in a Gemini® C18 column with a gradient of 0.1% formic acid in water and 0.1% formic acid in acetonitrile, at a flow rate of 250 μL/min and with a total run time of 9 min. The mass spectrometer was operated in multiple reaction monitoring (MRM) by monitoring citalopram transition 325.3→109.0, and internal standard transition 267.3→72.2 for quantification. The qualifier transitions 325.3→83.1 and 267.3→190.8, respectively, were also monitored. Linearity was observed from 32.4 to 973.2 ng/mL and the limit of quantitation achieved was 32.4 ng/mL. Also, the intermediate precision, repeatability and accuracy were below the acceptance limits of 15%. This method was applied to plasma and hair samples that were collected from mice submitted to a treatment with citalopram for different days. The plasma concentration–time profile of citalopram showed a tendency to stabilize, approaching zero as samples were collected 24 hours after the last drug administration. In contrast, the concentration-time profile in hair increased over the period of 30 days.
Highlights
Citalopram (1-[3-(Dimethylamino)propyl]-1-(4-fluorophenyl)1,3-dihydro-5-iso-benzofuran carbonitrile) is considered one of the most prescribed drugs being a selective and potent serotonin reuptake inhibitor [1]
We developed and validated a method for the determination and quantification of citalopram in mice plasma and hair with a small quantity of sample and with a simple extraction procedure, in contrast to the current literature which is mostly based on methods for humans
Selectivity: The selectivity of the method was evaluated by analyzing blank samples of plasma and hair from six different animals to investigate the potential interferences at the peak region for citalopram and IS
Summary
Citalopram (1-[3-(Dimethylamino)propyl]-1-(4-fluorophenyl)1,3-dihydro-5-iso-benzofuran carbonitrile) is considered one of the most prescribed drugs being a selective and potent serotonin reuptake inhibitor [1]. The small sample amount used for citalopram analysis in mice is significant for studies using this animal model This method for both matrices was successfully used to characterize the time course of changes in plasma and hair concentrations of citalopram following intraperitoneal injection for different periods of time in mice. Followed by centrifugation at 12,000×ɡ for 2 minutes, the plasma was recovered to another tube (BD Microtainer®, Becton Dickinson) with protease and phosphatase inhibitors (Roche) Regarding hair samples, they were pulled out with tweezers and placed into a microcentrifuge tube. 210 μL of MeOH (three times the plasma volume) were added and the samples were rehomogenized, followed by continuous agitation for 5 minutes at 1000 rpm’s in a thermomixer (Comfort, Eppendorf®).
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