HPLC analyses were performed as described before, using an 1100 series HPLC system (Agilent) with a Luna 5μ C18 reversed-phase column (Phenomenex) and 50% acetone and 50% 20 mm formic acid as the mobile phase (6 (link)).
Luna 5μ c18 reversed phase column
Manufactured by Phenomenex
Sourced in United Kingdom, United States
The Luna 5μ C18 reversed-phase column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. It features a 5 μm particle size and a C18 stationary phase, which is a commonly used sorbent in reversed-phase chromatography. The column is suitable for a variety of applications, including the analysis of pharmaceuticals, environmental samples, and other complex mixtures.
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Lab products found in correlation
3 protocols using luna 5μ c18 reversed phase column
1
Reversed-Phase HPLC for Compound Analysis
2
GAG Disaccharide Composition Analysis
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GAG samples were dissolved in 100 mM sodium acetate, 0.5 mM CaCl2, pH 7.1 and HS was exhaustively digested into disaccharides by incubation with heparinase I (10 mU, Grampian enzymes, Orkney, UK) overnight at 30°C, followed by a second incubation with heparinase II and heparinase III (10 mU each, Grampian enzymes) for 24 h at 37°C. Compositional analysis was performed by RPIP-HPLC, as described previously [31 (link)]. Samples were applied to a Luna 5μ C18 reversed phase column (4.6 × 150 mm, Phenomenex) equilibrated at 0.5 mL/min in 1.2 mM tetra-N-butylammonium hydrogen sulfate and 8.5% acetonitrile, and then resolved using a NaCl gradient (0–8 mM in 10 min, 8–30 mM in 1 min, 30–56 mM in 11.5 min, 56–106 mM in 1.5 min, and 106 mM for 6 min) calibrated with disaccharide standards (Iduron, Alderley Edge, UK). On-line post-column disaccharide derivatization was achieved by the addition of 2-cyanoacetamide (0.25%) in NaOH (0.5%) at a flow rate of 0.16 mL/min, followed by fluorescence detection (excitation 346 nm, emission 410 nm). Disaccharide analyses of each pool were performed in triplicate.
3
HPLC Quantification of Neurotransmitter Amino Acids
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The quantitative analysis of the neurotransmitter amino acids (glutamate, aspartate, glutamine, GABA, glycine, and taurine) was performed by the high-performance liquid chromatography (HPLC) method of Márquez, Quesada, Sánchez-Jiménez, and De Castro (1986 (link)).
The HPLC system used in this study consisted of a Wellchrom Mini-star K-501 pump (Knauer, Germany), equipped with a column thermostat 5–85°C having a 20 μl loop injector (Knauer, Germany). The column used was a Luna 5μ C-18 reversed-phase column (5 μm particle size, 150 × 4.6 mm I.D.) supplied by Phenomenex (USA). The separated amino acids were detected by a Wellchrom spectrophotometer K-2600 with variable wavelength (Knauer, Germany). A chromatography workstation (Eurochrom 2000) was used to quantify the height of the amino acids. The mobile phase consisted of methanol and water (50/50 v/v), to which glacial acetic acid (0.6%) and triethylamine (0.008%) were added. The height ratio of each amino acid was quantified (height of amino acid divided by the height of internal standard) and the concentrations (μmol/g fresh tissue) were then calculated by the internal standard method.
The HPLC system used in this study consisted of a Wellchrom Mini-star K-501 pump (Knauer, Germany), equipped with a column thermostat 5–85°C having a 20 μl loop injector (Knauer, Germany). The column used was a Luna 5μ C-18 reversed-phase column (5 μm particle size, 150 × 4.6 mm I.D.) supplied by Phenomenex (USA). The separated amino acids were detected by a Wellchrom spectrophotometer K-2600 with variable wavelength (Knauer, Germany). A chromatography workstation (Eurochrom 2000) was used to quantify the height of the amino acids. The mobile phase consisted of methanol and water (50/50 v/v), to which glacial acetic acid (0.6%) and triethylamine (0.008%) were added. The height ratio of each amino acid was quantified (height of amino acid divided by the height of internal standard) and the concentrations (μmol/g fresh tissue) were then calculated by the internal standard method.
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