Correlation of 3-mercaptopropionic acid induced seizures and changes in striatal neurotransmitters monitored by microdialysis.

OBJECTIVES: The goal of this study was to use a status epilepticus steady-state chemical model in rats using the convulsant, 3-mercaptopropionic acid (3-MPA), and to compare the changes in striatal neurotransmission on a slow (5min) and fast (60s) timescale. In vivo microdialysis was combined with electrophysiological methods in order to provide a complete evaluation of the dynamics of the results obtained. OBJECTIVE: To compare the effects of a steady-state chemical model pof status epilepticus on striatal amino-acid and amine neurotransmitters contents, as measured via in vivo microdialysis combined with electrophysiological methods. Measurements were performed on samples collected every 60s and every 5min. "Fast" (60s) and "slow" (5min) sampling timescales were selected, to gain more insight into the dynamics of GABA synthesis inhibition and of its effects on other neurotransmitters and on cortical electrical activity. METHODS: 3-MPA was administered in the form of an intra-venous load (60mg/kg) followed by a constant infusion (50mg/kg/min) for min. Microdialysis samples were collected from the striatum at intervals of 5min and 60s and analyzed for biogenic amine and amino acid neurotransmitters. ECoG activity was monitored via screws placed over the cortex. RESULTS: In the 5min samples, glutamate (Glu) increased and γ-aminobutyric acid (GABA) decreased monotonically while changes in dopamine (DA) concentration were bimodal. In the sixty second samples, Glu changes were bimodal, a feature that was not apparent with the 5min samples. ECoG activity was indicative of status epilepticus. CONCLUSIONS: This study describes the combination of in vivo microdialysis with electrophysiology to monitor the effect of 3-MPA on neurotransmission in the brain. This led to a better understanding of the chemical changes in the striatum due to the applied 3-MPA chemical model of status epilepticus.

A parallel dual-electrode detector for capillary electrophoresis.

An approach to on-capillary dual-electrode detection for CE using a parallel electrode configuration has been developed. The parallel configuration provides two operating modes. In the first mode, one working electrode is held at an oxidizing potential and the second working electrode is held at a reducing potential. This results in redox cycling of analytes between the oxidized and reduced forms, enhancing sensitivity compared to single-electrode detection. In the second mode, both working electrodes are held at different oxidizing potentials. This mode provides electrochemical characterization of electrophoretic peaks. In the redox cyclying mode, signal enhancement of up to twofold was observed for the dual-electrode detection of phenolic acid standards compared to single-electrode detection. Variation in response of less than 10% from electrode to electrode was determined (at a concentration of 60 nM) indicating reproducible fabrication. LODs were determined to be as low as 5.0 nM for dual-electrode configuration. Using the dual-potential mode peak identification of targeted phenolic acids in whiskey samples were confirmed based on both migration time and current ratios.

An investigation into the pharmacokinetics of 3-mercaptopropionic acid and development of a steady-state chemical seizure model using in vivo microdialysis and electrophysiological monitoring.

OBJECTIVES: The goal of the present study was to develop a chemical seizure model using the convulsant, 3-mercaptopropionic acid (3-MPA). A pharmacodynamics approach was taken, combining in vivo microdialysis sampling with electrophysiological methods to simultaneously monitor, in real-time, the 3-MPA concentration in the brain and the corresponding electrocorticographic (ECoG) activity. METHODS: The 3-MPA was administered in two doses (50 and 100 mg/kg) in order to study its pharmacokinetics. Microdialysis samples were collected from the striatum, hippocampus, and jugular vein every 5 min. The microdialysates were analyzed using high-performance liquid chromatography with electrochemical detection (HPLC-EC). The ECoG activity was monitored via screws placed onto the cortex. Noncompartmental pharmacokinetics analysis was performed to obtain the elimination constants (K(e)), the maximum concentration (C(max)), the time to achieve maximum concentration (T(max)), and the area under the concentration-time curves (AUC(inf)). RESULTS: The average brain K(e) for the 50 and the 100mg/kg doses were 0.060 and 0.018 min(-1), respectively. The brain AUC(inf) for the 50 and 100mg/kg doses were 353 and 2168 mg min(-1)mL(-1), respectively. This led to a 67-fold increase in the observed number of seizures in the higher dose with the average seizure intensity double that of the smaller dose. These data led to the dosing scheme for the chemical seizure model of administering a 3-MPA loading dose of 60 mg/kg followed by a constant infusion of 50 mg/(kg min(-1)). CONCLUSIONS: This study describes, to our knowledge, the first successful attempt to combine in vivo microdialysis with electrophysiology to monitor in real-time, the concentration and effects of 3-MPA in the brain. This led to the development of a steady-state chemical seizure model.

Solid-phase extraction sorbent consisting of alkyltrimethylammonium surfactants immobilized onto strong cation-exchange polystyrene resin.

Presented is a solid-phase extraction sorbent material composed of cationic alkyltrimethylammonium surfactants attached to a strong cation-exchange resin via ion-exchange. The original hydrophilic cation-exchange resin is made hydrophobic by covering the surface with alkyl chains from the hydrophobic portion of the surfactant. The sorbent material now has a better ability to extract hydrophobic molecules from aqueous samples. The entire stationary phase (alkyltrimethylammonium surfactant) is removed along with the analyte during the elution step. The elution step requires a mild elution solvent consisting of 0.25 M Mg2+ in a 50% 2-propanol solution. The main advantage of using a removable stationary phase is that traditionally utilized toxic elution solvents such as methylene chloride, which are necessary to efficiently release strongly hydrophobic species from SPE stationary phases, may now be avoided. Also, the final extract is directly compatible with reversed-phase liquid chromatography. The performance of this procedure is presented using pyrene as a test molecule.