Microchip electrophoresis with electrochemical detection for the determination of analytes in the dopamine metabolic pathway.

Electrophoresis (2015-05-12)
Rachel A Saylor, Erin A Reid, Susan M Lunte
RESUMEN

A method for the separation and detection of analytes in the dopamine metabolic pathway was developed using microchip electrophoresis with electrochemical detection. The microchip consisted of a 5 cm PDMS separation channel in a simple-t configuration. Analytes in the dopamine metabolic pathway were separated using a background electrolyte composed of 15 mM phosphate at pH 7.4, 15 mM SDS, and 2.5 mM boric acid. Two different microchip substrates using different electrode materials were compared for the analysis: a PDMS/PDMS device with a carbon fiber electrode and a PDMS/glass hybrid device with a pyrolyzed photoresist film carbon electrode. While the PDMS/PDMS device generated high separation efficiencies and good resolution, more reproducible migration times were obtained with the PDMS/glass hybrid device, making it a better choice for biological applications. Lastly, the optimized method was used to monitor l-DOPA metabolism in a rat brain slice.

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Sigma-Aldrich
2-Propanol, BioReagent, for molecular biology, ≥99.5%
Sigma-Aldrich
Sodium dodecyl sulfate, BioReagent, suitable for electrophoresis, for molecular biology, ≥98.5% (GC)
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2-Propanol, anhydrous, 99.5%
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Sodium dodecyl sulfate, ACS reagent, ≥99.0%
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Dopamine hydrochloride
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Sodium phosphate monobasic, ReagentPlus®, ≥99.0%
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Sodium hydroxide solution, BioUltra, for molecular biology, 10 M in H2O
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Sodium hydroxide solution, 1.0 N, BioReagent, suitable for cell culture
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Sodium dodecyl sulfate, ReagentPlus®, ≥98.5% (GC)
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Boric acid, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
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Isopropyl alcohol, ≥99.7%, FCC, FG
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Sodium phosphate dibasic, ReagentPlus®, ≥99.0%
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Sodium dodecyl sulfate solution, BioUltra, for molecular biology, 10% in H2O
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Sodium dodecyl sulfate, ≥99.0% (GC), dust-free pellets
Sigma-Aldrich
Sodium dodecyl sulfate, BioUltra, for molecular biology, ≥99.0% (GC)
Sigma-Aldrich
Sodium phosphate dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium phosphate dibasic, for molecular biology, ≥98.5% (titration)
Sigma-Aldrich
2-Propanol, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
Sodium dodecyl sulfate solution, BioUltra, for molecular biology, 20% in H2O
Sigma-Aldrich
Sodium phosphate monobasic, BioReagent, for molecular biology, anhydrous, ≥98%
Sigma-Aldrich
Sodium phosphate monobasic, BioXtra, ≥99.0%
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Sodium dodecyl sulfate, BioXtra, ≥99.0% (GC)
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Sodium phosphate dibasic solution, BioUltra, 0.5 M in H2O
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Sodium dodecyl sulfate, 92.5-100.5% based on total alkyl sulfate content basis
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Sodium phosphate monobasic, BioPerformance Certified, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0% (titration)
Supelco
Sodium dodecyl sulfate, dust-free pellets, suitable for electrophoresis, for molecular biology, ≥99.0% (GC)
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Sodium phosphate monobasic solution, BioUltra, 5 M in H2O
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2-Propanol, electronic grade, 99.999% trace metals basis
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Sodium phosphate monobasic, meets USP testing specifications, anhydrous