Ecd 3000rs

Manufactured by Thermo Fisher Scientific

Sourced in United States

The ECD-3000RS is a laboratory instrument designed for the detection and measurement of specific chemical compounds. It utilizes an electron capture detector (ECD) technology to provide sensitive and selective analysis capabilities. The core function of the ECD-3000RS is to detect and quantify the presence of certain target analytes in various sample matrices.

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Lab products found in correlation

Hr 80 reverse phase column, by Thermo Fisher Scientific (1 mentions) Chromeleon 7.2 chromatography data system, by Thermo Fisher Scientific (1 mentions) Dionex ultimate 3000, by Thermo Fisher Scientific (1 mentions) Chromeleon 7, by Thermo Fisher Scientific (1 mentions) Betasil c18, by Thermo Fisher Scientific (1 mentions) Md 150 3, by Thermo Fisher Scientific (1 mentions) Wps 3000tbrs, by Thermo Fisher Scientific (1 mentions)

4 protocols using ecd 3000rs

Catecholamine Quantification in Plasma and Tissue

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Plasma and tissue catecholamine content was measured using a reverse-phase HPLC system (Thermo UltiMate^TM 3000 UHPLC, Thermo Fisher Scientific, Waltham, MA, USA) with electrochemical detection (ECD-3000RS, Thermo Fisher Scientific), following an established protocol [15 ,21 (link)] modified from the method of Mefford [22 (link)]. Briefly, 100 µL of plasma or ~100 mg of tissue was homogenized in 0.2 M perchloric acid with 1 mg/ml ascorbic acid solution containing the internal standard dihydroxybenzylamine (DHBA, Thermo Fisher Scientific) to control extraction efficiency. After centrifugation, catecholamines were extracted from the homogenate with alumina and 0.5 M tris(hydroxymethyl)aminomethane buffer (pH 8.6) and were eluted into the perchloric acid with ascorbic acid solution. The extracted catecholamines were assayed with Cat-A-Phase II mobile phase (Thermo Fisher Scientific) and separated with a HR-80 reverse phase column (Thermo Fisher Scientific). The electrochemical detector settings were: guard cell, +250 mV; cell 1, +100 mV; and cell 2, −200 mV. Catecholamine values were calculated based on the standard curve from each assay and were corrected for extraction efficiency based on the internal standard DHBA within each sample.

Zhu Q., Liu X., Glazier B.J., Krolick K.N., Yang S., He J., Lo C.C, & Shi H. (2019). Differential Sympathetic Activation of Adipose Tissues by Brain-Derived Neurotrophic Factor. Biomolecules, 9(9), 452.

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Quantitative Analysis of Dopamine using UHPLC-ECD

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The detection and quantification of dopamine was performed in a similar protocol as previously described [26 (link)]. Prior to dialysate analysis, a standard curve consisted of a series of known dopamine concentrations was established for every experiment. The dopamine assay was performed on UHPLC system (Thermo Scientific™ Dionex™ UltiMate™ 3000) equipped with ECD-3000RS (Electrochemical cell: 6011RS, Thermo Scientific). Samples were injected automatically and separated on an analytical column Acclaim RSLC PA2, 250 × 2.1 mm, 2.2 μm (Thermo Scientific, 074814). For dopamine assay, the applied potential for analytical cell was set as + 220 mV. The UHPLC system was operated at 400 μL/min for 10 mins at 30 °C using test mobile phase (Thermo Scientific). The column temperature was set to 30 °C. The concentration of dopamine was measured relative to standard solutions using Chromeleon 7.2 Chromatography Data System (Dionex, Thermo Scientific).

Lai T.K., Su P., Zhang H, & Liu F. (2018). Development of a peptide targeting dopamine transporter to improve ADHD-like deficits. Molecular Brain, 11, 66.

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Quantifying Coenzyme Q Levels

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Extracted lipids were resuspended in 2-propanol (50 μl) and transferred to an amber vial. Sodium borohydride (2 μl of 10 mM in 2-propanol) was added to each vial, followed by brief vortexing and incubation (10 min, r.t.) to reduce the CoQ. Methanol (50 μl) was then added to each sample to remove excess sodium borohydride and the vials were flushed with argon gas. CoQ measurements were performed using reverse-phase HPLC-ECD66 . Separation was performed using a C18 column (Thermo Scientific, Betasil C18, 100 × 2.1 mm, particle size 3 μm) at a flow rate of 0.3 ml min⁻¹ with a mobile phase of 78% methanol, 10% 2-propanol, 10% acetonitrile and 2% ammonium acetate (1 M, pH 4.4). Electrochemical detection was performed using an ECD detector (Thermo Scientific ECD3000-RS) containing a 6020RS omni Coulometric Guarding Cell (E1) set to −200 mV and two 6011RS ultra Analytical Cells (E2 and E3) set to 600 mV. CoQ measurements were made on the analytical E2 electrode. For each experiment, CoQ₆ and CoQ₈ standards (Avanti) were prepared in the same manner as the experimental samples. Peak areas were quantified using Chromeleon 7.2.10 software (Thermo) and normalized to the CoQ₈ internal standard. CoQ₆ levels were further normalized to OD₆₀₀ at collection.

Tai J., Guerra R.M., Rogers S.W., Fang Z., Muehlbauer L.K., Shishkova E., Overmyer K.A., Coon J.J, & Pagliarini D.J. (2023). Hem25p is required for mitochondrial IPP transport in fungi. Nature cell biology, 25(11), 1616-1624.

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Dopamine Quantification in Microdissected Brain Regions

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HPLC on microdissected dopaminergic brain regions was carried out to assess whether tissular DA levels differed between RGS12-null and wild-type mice. Dorsal and ventral striatal brain regions were rapidly dissected and frozen on dry ice. Tissue was then sonicated in 10 volumes of 0.3 N perchloric acid and homogenates were centrifuged at 12,000×g for 15 min. Supernatants were collected, passed through a 0.22 μm filter, and dopamine levels in the homogenates were determined by HPLC with dual-cell electrochemical detection. Cleared lysates underwent automated direct injection of 10 μL into a Dionex Ultimate 3000 HPLC system equipped with a pump (ISO-3100BM, Thermo Fisher Scientific, Waltham, Massachusetts, USA) linked to an autosampler (WPS-3000TBRS, Thermo Fisher Scientific, Waltham, MA, USA), ESA HPLC column (MD-150×3.2) and dual coulometric/amperometric electrochemical detectors (ECD-3000RS, Thermo Fisher Scientific, Waltham, Massachusetts, USA) set at −170 mV and +400 mV, respectively. Mobile phase (MD-TM; Thermo Fisher Scientific, Waltham, Massachusetts, USA) was pumped at a flow rate of 0.6 mL/min. Chromatograms were analyzed using Chromeleon software and dopamine peak height (concentration) was determined from a standard curve. The identity of the DA peak in the sample was confirmed by standard addition.

Gross J.D., Kaski S.W., Schroer A.B., Wix K.A., Siderovski D.P, & Setola V. (2018). Regulator of G protein signaling-12 modulates the dopamine transporter in ventral striatum and locomotor responses to psychostimulants. Journal of psychopharmacology (Oxford, England), 32(2), 191-203.

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