Optimization of ultra-performance liquid chromatography (UPLC) with fluorescence detector (FLD) method for the quantitative determination of selected neurotransmitters in rat brain

• Authors: Joanna Stragierowicz , Adam Daragó , Sławomir Brzeźnicki , Anna Kilanowicz

Abstracts

EN

Background Glutamate (Glu) and γ-aminobutyric acid (GABA) are the main neurotransmitters in the central nervous system for excitatory and inhibitory processes, respectively. Monitoring these neurotransmitters is an essential tool in establishing pathological functions, among others in terms of occupational exposure to toxic substances. Material and Methods We present modification of the HPLC (high-performance liquid chromatography) to the UPLC (ultra-performance liquid chromatography) method for the simultaneous determination of glutamate and γ-aminobutyric acid in a single injection. The isocratic separation of these neurotransmitter derivatives was performed on Waters Acquity BEH (ethylene bridged hybrid) C18 column with particle size of 1.7 μm at 35°C using a mobile phase consisting of 0.1 M acetate buffer (pH 6.0) and methanol (60:40, v/v) at a flow rate of 0.3 ml/min. The analytes were detected with the fluorescence detector (FLD) using derivatization with o-phthaldialdehyde (OPA), resulting in excitation at 340 nm and emission at 455 nm. Results Several validation parameters including linearity (0.999), accuracy (101.1%), intra-day precision (1.52–1.84%), inter-day precision (2.47–3.12%), limit of detection (5–30 ng/ml) and quantification (100 ng/ml) were examined. The developed method was also used for the determination of these neurotransmitters in homogenates of selected rat brain structures. Conclusions The presented UPLC-FLD is characterized by shorter separation time (3.5 min), which is an adaptation of the similar HPLC methods and is an alternative for more expensive references techniques such as liquid chromatography coupled with tandem mass-spectrometry (LC-MS/MS) methods. Med Pr 2017;68(5):583–591

Keywords

EN

brain; neurotransmitters; glutamate; fluorescence detector; ultra-performance liquid chromatography; γ-aminobutyric acid

• Publisher: Instytut Medycyny Pracy im. prof. dra Jerzego Nofera w Łodzi
• Journal: Medycyna Pracy
• Year: 2017
• Volume: 68
• Issue: 5
• Pages: 583-591

Dates

published

2017

Contributors

author

Joanna Stragierowicz

• Medical University of Lodz, Łódź, Poland (Faculty of Pharmacy, Department of Toxicology)

author

Adam Daragó

• Medical University of Lodz, Łódź, Poland (Faculty of Pharmacy, Department of Toxicology)

author

Sławomir Brzeźnicki

• Nofer Institute of Occupational Medicine, Łódź, Poland (Department of Biological and Environmental Monitoring)

author

Anna Kilanowicz

• Medical University of Lodz, Łódź, Poland (Faculty of Pharmacy, Department of Toxicology)

Identifiers

DOI

10.13075/mp.5893.00622

Biblioteka Nauki

2164059