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

2.50
Hdl Handle:
http://hdl.handle.net/10146/618116
Title:
Optimization of ultra-performance liquid chromatography (UPLC) with fluorescence detector (FLD) method for the quantitative determination of selected neurotransmitters in rat brain.
Authors:
Stragierowicz, Joanna; Darago, Adam; Brzeznicki, Slawomir ( 0000-0002-0542-8538 ) ; Kilanowicz, Anna
Abstract:
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.; 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.; 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.; 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.
Affiliation:
Nofer Institute of Occupational Medicine, Lodz, Poland
Citation:
Medycyna Pracy 2017, 68 (5):583-591
Journal:
Medycyna pracy
Issue Date:
26-Jul-2017
URI:
http://hdl.handle.net/10146/618116
DOI:
10.13075/mp.5893.00622
PubMed ID:
28749489
Additional Links:
http://medpr.imp.lodz.pl/Optimization-of-ultra-performance-liquid-chromatography-UPLC-with-fluorescence-detector-FLD-method-for-the-quantitative-determination-of-selected-neurotransmitters-in-rat-brain,74312,0,2.html
Type:
Article
Language:
en
ISSN:
0465-5893
Sponsors:
Funding: the work is financially supported by the Medical University of Lodz, project No. 502-03/3-045-01/502-34-043 entitled “Evaluation of the neurotoxicity of hexachloronaphthalene in the subchronic study in rats.” Project manager: Joanna Stragierowicz, M.Sc.
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DC FieldValue Language
dc.contributor.authorStragierowicz, Joannaen
dc.contributor.authorDarago, Adamen
dc.contributor.authorBrzeznicki, Slawomiren
dc.contributor.authorKilanowicz, Annaen
dc.date.accessioned2017-08-21T09:34:22Z-
dc.date.available2017-08-21T09:34:22Z-
dc.date.issued2017-07-26-
dc.identifier.citationMedycyna Pracy 2017, 68 (5):583-591en
dc.identifier.issn0465-5893-
dc.identifier.pmid28749489-
dc.identifier.doi10.13075/mp.5893.00622-
dc.identifier.urihttp://hdl.handle.net/10146/618116-
dc.description.abstractGlutamate (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.en
dc.description.abstractWe 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.en
dc.description.abstractSeveral 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.en
dc.description.abstractThe 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.en
dc.description.sponsorshipFunding: the work is financially supported by the Medical University of Lodz, project No. 502-03/3-045-01/502-34-043 entitled “Evaluation of the neurotoxicity of hexachloronaphthalene in the subchronic study in rats.” Project manager: Joanna Stragierowicz, M.Sc.en
dc.language.isoenen
dc.relation.urlhttp://medpr.imp.lodz.pl/Optimization-of-ultra-performance-liquid-chromatography-UPLC-with-fluorescence-detector-FLD-method-for-the-quantitative-determination-of-selected-neurotransmitters-in-rat-brain,74312,0,2.htmlen
dc.rightsArchived with thanks to Medycyna pracyen
dc.subjectBrainen
dc.subjectglutamateen
dc.subjectneurotransmittersen
dc.subjectfluorescence detectoren
dc.subjectultra-performance liquid chromatographyen
dc.subjectγ-aminobutyric aciden
dc.titleOptimization of ultra-performance liquid chromatography (UPLC) with fluorescence detector (FLD) method for the quantitative determination of selected neurotransmitters in rat brain.en
dc.typeArticleen
dc.contributor.departmentNofer Institute of Occupational Medicine, Lodz, Polanden
dc.identifier.journalMedycyna pracyen
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