An isocratic high performance liquid chromatography method for the determination of GABA and glutamate in discrete regions of the rodent brain

Clarke, Gerard; O’Mahony, Siobhain M.; Malone, Gwen; Dinan, Timothy G.

doi:10.1016/j.jneumeth.2006.09.006

Cited by 68 publications

(47 citation statements)

References 24 publications

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“…However, in some studies the run time remained the same [4,14]. In addition, the chromatograms of the GABA derivatives using OPA or other derivatizing agents showed prominent peaks of these derivatizing agents which hindered the resolution of the amino acids under study [11,12,19]. The recent methods documented on the estimation of GABA by several modifications of the existing methods through modification of mobile phase composition, pH adjustments, changing the flow rate, column temperature, or the substitution of the conventional derivatizing agents also resulted in an increase in the runtime for the analysis of GABA which ranged from 10 to 60 min [3,4,12].…”

Section: Chromatographic Estimationmentioning

confidence: 99%

“…Use of several other derivatizing agents like dansyl chloride, naphthalene-2,3-dicarboxaldehyde, fluorescamine, phenyl isothiocyanate, 5-(4,6-dichlorostriazin-2-ylamino) fluorescein, benzylamine with 1,2-diphenylethylenediamine, etc. has also been documented but those methods also carry stability issues and longer run times [4,[9][10][11]. The reduced efficiency in GABA analysis with existing gradient elution methods, enhanced run times, tedious analytical procedure, and poor resolution due to interference from derivatizing agents themselves and other matrix components have led to the optimization of the chromatographic methods to ensure the detection of GABA with good resolution [3,4,7,8,12].…”

Section: Introductionmentioning

confidence: 99%

“…The reduced efficiency in GABA analysis with existing gradient elution methods, enhanced run times, tedious analytical procedure, and poor resolution due to interference from derivatizing agents themselves and other matrix components have led to the optimization of the chromatographic methods to ensure the detection of GABA with good resolution [3,4,7,8,12]. The modern methods also make the chromatographic estimation more expensive with the use of costly derivatizing agents like OPA and intricate technical requirements using gradient pumps, mass spectroscopic identification, microfluidic electrophoresis chip, microchip electrophoresis, and monolithic column chromatography coupled with chemiluminescence detection [4,10,11,13,14].…”

Section: Introductionmentioning

confidence: 99%

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Development and Validation of a Specific RP-HPLC Method for the Estimation of γ-Aminobutyric Acid in Rat Brain Tissue Samples Using Benzoyl Chloride Derivatization and PDA Detection

Akula¹,

Chandrasekaran²,

Kaur³

et al. 2015

Acta Chromatographica

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Summary.A new, rapid, and specific reversed phase high-performance liquid chromatographic (RP-HPLC) method involving precolumn derivatization with benzoyl chloride was developed and validated for the estimation of γ-aminobutyric acid (GABA) in rat brain tissue preparations. The derivatization product of GABA was identified by melting point, infrared, and proton nuclear magnetic resonance ( 1 H NMR) spectroscopy to be n-benzoyl GABA. Various parameters which influenced derivatization and elusion were optimized. The chromatographic system consisted of C-18 column with ultraviolet (UV)-photodiode array detection ranging from 210 to 400 nm. Elution with an isocratic mobile phase consisting of 0.025 M disodium hydrogen phosphate buffer-methanol (65:35, v/v; pH 6) at a flow rate of 1 mL min −1 yielded sharp and specific peak of n-benzoyl GABA within 7 min. The method was validated with respect to the linearity, accuracy, precision, sensitivity, selectivity, and stability, wherein the benzoyl derivative of GABA showed stability for 2 months. The lower limit of detection was 0.5 nmol L −1 . This novel derivatization procedure for the estimation of GABA with benzoyl chloride was also applied for rat brain tissue preparations that gave highly specific peak and good component recovery. The results show that the method for the determination of GABA by benzoylation using RP-HPLC has good linearity, accuracy, precision, sensitivity, and specificity and is simple and economical to perform.

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Section: Chromatographic Estimationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development and Validation of a Specific RP-HPLC Method for the Estimation of γ-Aminobutyric Acid in Rat Brain Tissue Samples Using Benzoyl Chloride Derivatization and PDA Detection

Akula¹,

Chandrasekaran²,

Kaur³

et al. 2015

Acta Chromatographica

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“…Sample analysis was carried out by HPLC coupled to fluorescence detection as described previously (Clarke et al, 2007). Briefly, samples were sonicated for 4 s (Sonoplus; Bandelin) in chilled homogenization buffer (0.1 M citric acid, 0.1 M sodium dihydrogen phosphate monohydrate, 5.6 mM octane sulfonic acid, 10 mM EDTA in 10% (v/v) methanol solution (pH 2.8) with 4 M NaOH).…”

Section: Hplc Sample Analysismentioning

confidence: 99%

A Sensitive Period of Mice Inhibitory System to Neonatal GABA Enhancement by Vigabatrin is Brain Region Dependent

Levav-Rabkin

Melamed

Clarke

et al. 2009

Neuropsychopharmacol

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Neurodevelopmental disorders, such as schizophrenia and autism, have been associated with disturbances of the GABAergic system in the brain. We examined immediate and long-lasting influences of exposure to the GABA-potentiating drug vigabatrin (GVG) on the GABAergic system in the hippocampus and cerebral cortex, before and during the developmental switch in GABA function (postnatal days P1-7 and P4-14). GVG induced a transient elevation of GABA levels. A feedback response to GABA enhancement was evident by a short-term decrease in glutamate decarboxylase (GAD) 65 and 67 levels. However, the number of GAD65/67-immunoreactive (IR) cells was greater in 2-week-old GVG-treated mice. A long-term increase in GAD65 and GAD67 levels was dependent on brain region and treatment period. Vesicular GABA transporter was insensitive to GVG. The overall effect of GVG on the Cl À co-transporters NKCC1 and KCC2 was an enhancement of their synthesis, which was dependent on the treatment period and brain region studied. In addition, a short-term increase was followed by a long-term decrease in KCC2 oligomerization in the cell membrane of P4-14 hippocampi and cerebral cortices. Analysis of the Ca 2 + binding proteins expressed in subpopulations of GABAergic cells, parvalbumin and calbindin, showed region-specific effects of GVG during P4-14 on parvalbumin-IR cell density. Moreover, calbindin levels were elevated in GVG mice compared to controls during this period. Cumulatively, these results suggest a particular susceptibility of the hippocampus to GVG when exposed during days P4-14. In conclusion, our studies have identified modifications of key components in the inhibitory system during a critical developmental period. These findings provide novel insights into the deleterious consequences observed in children following prenatal and neonatal exposure to GABA-potentiating drugs.

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“…So it is very necessary to develop a rapid and sensitive method to detect L-glutamate in biological and food samples. So far, much efforts have been devoted to develop detection methods of L-glutamate, such as high performance liquid chromatography (HPLC) [20], capillary electrophoresis [21,22], optical methods [23][24][25] and electrochemical analyses [26,27]. Among these approaches, electrochemical methods, especially electrochemical biosensors, have attracted much attention due to their high sensitivity, rapid response and easy operation [28][29][30][31].…”

Section: Introductionmentioning

confidence: 99%

Amperometric l-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase

Liang

Zhang

Lang

et al. 2015

Analytica Chimica Acta

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An isocratic high performance liquid chromatography method for the determination of GABA and glutamate in discrete regions of the rodent brain

Cited by 68 publications

References 24 publications

Development and Validation of a Specific RP-HPLC Method for the Estimation of γ-Aminobutyric Acid in Rat Brain Tissue Samples Using Benzoyl Chloride Derivatization and PDA Detection

Development and Validation of a Specific RP-HPLC Method for the Estimation of γ-Aminobutyric Acid in Rat Brain Tissue Samples Using Benzoyl Chloride Derivatization and PDA Detection

A Sensitive Period of Mice Inhibitory System to Neonatal GABA Enhancement by Vigabatrin is Brain Region Dependent

Amperometric l-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase

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