Determination of in vivo amino acid neurotransmitters by high-performance liquid chromatography with o-phthalaldehyde-sulphite derivatisation

Rowley, Helen L.; Martin, Keith F.; Marsden, C.A.

doi:10.1016/0165-0270(94)00132-z

Cited by 208 publications

(124 citation statements)

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“…Overall, the two most attractive advantages of this method are higher stability of benzoyl derivative and convenience in analysis. The utility of isocratic mobile phase of disodium hydrogen phosphate buffer:methanol (65:35, v/v) and simple derivatization process reduced the chromatographic runtime below 7 min, indicating the higher efficiency of the present method as compared to the existing chromatographic methods for the detection of GABA, which yielded the run times up to 60 min [3,4] and with less stability of derivatives [7,3,12].…”

Section: Chromatographic Estimationmentioning

confidence: 97%

“…However, the products formed using these derivatizing agents are not stable for longer duration and require faster online estimation [7]. The half-lives of various GABA derivatives are compared in Table II.…”

Section: Chromatographic Estimationmentioning

confidence: 99%

“…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]. 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%

“…In order to circumvent these limitations with the estimation of GABA, a precolumn derivatization coupled with HPLC-fluorescence/electrochemical detection has been in use for the detection and/or quantification of this amino acid in microdialysate samples, brain tissue extracts, and other biological matrix preparations. This precolumn derivatization was found to transform GABA into detectable forms, and therefore, derivatizing agents like o-phthalaldehyde (OPA) with β-mercaptoethanol or sodium sulphite have been widely used despite the poor stability of the electroactive and/or fluorescent derivatives [4][5][6][7][8]. 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.…”

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: 97%

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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“…Glutamate levels in the extracellular fluids were measured electrochemically after derivatization with OPA/sulfite Microdialysis, tissue content, comet assay, caspase-3 activity, and Ht1a, Ht2a, Gpx3, Sod1 expression 1 3 reagent to form isoindole-sulfonate derivative [25]. Chromatography was performed using an Ultimate 3000 pump (Dionex), LC-4B amperometric detector with a cross-flow detector cell (BAS) and an HR-80 column (80 × 4.6 mm, particle size 3 μm; ESA Inc, Chelmsford, MA, USA).…”

Section: Extracellular Concentrations Of Da 5-ht and Glutamatementioning

confidence: 99%

Effects of exposure to 5-MeO-DIPT during adolescence on brain neurotransmission and neurotoxicity in adult rats

et al. 2018

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Purpose Tryptamine hallucinogen 5-methoxy-N,N-diisopropyltryptamine (5-MeO-DIPT) is a serotonin transporter inhibitor with high affinity for serotonin 5-HT 1A and 5-HT 2A/C receptors. We showed previously that 5-MeO-DIPT in a single dose increased neurotransmitter release in brain regions of rats and elicited single-and double-strand DNA breaks. Herein we investigated the effects of repeated-intermittent 5-MeO-DIPT administration in adolescence on dopamine (DA), serotonin (5-HT) and glutamate release in brain regions of adult rats. Furthermore, we examined caspase-3 activity, oxidative DNA damage, the Gpx3, Sod1, Ht1a and Ht2a mRNA expression levels, and cell viability. Methods Neurotransmitter release was measured by microdialysis in freely moving animals. Caspase-3 activity was assessed colorimetrically, and oxidative DNA damage with the comet assay, while the Gpx3, Sod1, Ht1a and Ht2a mRNA expression levels were assessed by real-time polymerase chain reaction. Cell viability was studied in SH-SY5Y and Hep G2 cells by the MTT test. Results We observed changed responses of DA, 5-HT and glutamate neurons to a challenge dose of 5-MeO-DIPT when animals were treated repeatedly in adolescence with this hallucinogen. The basal extracellular levels of DA and 5-HT were decreased in the striatum and nucleus accumbens, while glutamate level was increased in the nucleus accumbens and frontal cortex. The damage of cortical DNA, increased Gpx3 and Sod1 mRNA expression and affected caspase-3 activity were also observed. Furthermore, decreased Ht1a and Ht2a mRNA expression in the frontal cortex and marked cytotoxicity of 5-MeO-DIPT were found. Conclusions These results suggest that 5-MeO-DIPT given repeatedly during adolescence affects brain neurotransmission and shows neurotoxic potential observed in adult animals.

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Voltammetry In Vivo for Chemical Analysis of the Living Brain

O’Neill

Lowry

2000

Encyclopedia of Analytical Chemistry

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The mammalian brain is the most complex structure known to science. Our behavior, feelings, thoughts, and maybe even consciousness itself, may be a reflection of the interplay of electrical and chemical signaling that create these states, and how the physical brain gives rise to the properties of mind remains a major unanswered question. It is clear, however, that many of the drugs used empirically in the treatment of neurological disorders, such as Parkinson's disease, schizophrenia and depression, as well as mind‐altering substances of abuse, have specific chemical actions on nerve cells in the brain, highlighting the importance of chemical signaling in the functioning of neural networks. A growing number of methodologies are being developed, including sampling, spectroscopic and electrochemical, to study neurochemical phenomena in the living brain. One such set of techniques focuses on the detection of substances released from nerve cells, using amperometric electrodes and voltammetry in vivo (VIV) techniques. By implanting a microvoltametric electrode in a specific brain region, applying a suitable potential profile and recording the resulting faradaic current, changes in the concentration of a variety of substances in the extracellular fluid (ECF) can be monitored with sub‐second time resolution over extended periods. This allows investigations of the functions and roles of specific neurochemicals in neuronal signaling, drug actions, and well‐defined behaviors. The main limitations associated with VIV is, on the one hand, the limited number of ECF species that are electroactive, and on the other, the fact that electroactive substances present tend to oxidize at similar potentials, interfering with each other's detection. These restrictions are being overcome by ongoing developments, including the design of biosensors to broaden the range of target analytes and permselective membranes to block interference.

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Determination of in vivo amino acid neurotransmitters by high-performance liquid chromatography with o-phthalaldehyde-sulphite derivatisation

Cited by 208 publications

References 29 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

Effects of exposure to 5-MeO-DIPT during adolescence on brain neurotransmission and neurotoxicity in adult rats

Voltammetry In Vivo for Chemical Analysis of the Living Brain

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