Tri carboxylic acid cycle and Glycolysis cycle metabolites were identified by using 5 mM of ammonium acetate in water as buffer PH 9.9 (A) and 100% acetonitrile as buffer (B) using the Luna 3 µM NH2 100 A0 Chromatography column (Phenomenex, Torrance, CA, USA). The gradient: 0–20 min—80% B (flow rate 0.2 mL/min); 20–20.10 min—80% to 2% B; 20.10–25 min—2% B (flow rate 0.3 mL/min); 25–30 min—80% B (Flowrate 0.35 mL/min); 30–35 min—80%B (Flow rate 0.4 mL/min); 35–38 min—80% B (flow rate 0.4 mL/min), followed by re-equilibration at the end of the gradient to the initial starting condition of 80% B at a flow rate of 0.2 mL/min. All the identified metabolites were normalized by the spiked internal standard.
Luna 3 m nh2 100 a0 chromatography column
Manufactured by Phenomenex
Sourced in United States
The Luna 3 μM NH2 100 Å Chromatography Column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of polar and ionic compounds. The column features a 3 μM particle size and a 100 Å pore size, which provides high-efficiency separation and good peak resolution. The column is packed with a chemically bonded aminopropyl stationary phase, which offers excellent selectivity for the analysis of polar analytes.
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2 protocols using luna 3 m nh2 100 a0 chromatography column
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Metabolite Profiling of TCA and Glycolysis
2
Metabolomic Analysis of TCA and Glycolysis
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Tricarboxylic acid cycle and Glycolysis cycle metabolites were identified by using 5 mM ammonium acetate in water as buffer PH 9.9 (A) and 100% acetonitrile as a buffer (B) using Luna 3 µM NH2 100 A0 Chromatography column (Phenomenex, Torrance, CA). The Gradient used: 0–20 min-80% B (Flow rate 0.2 ml/min); 20–20.10 min- 80% to 2% B; 20.10–25 min-2% B(Flow rate 0.3 ml/min); 25–30 min 80% B (Flowrate 0.35 ml/min); 30–35 min-80%B (Flow rate 0.4 ml/min); 35–38 min 80% B (Flow rate 0.4 ml/min); followed by re-equilibration at the end of the gradient to the initial starting condition 80% B a Flow rate of 0.2 ml/min. All the identified metabolites were normalized by spiked internal standard (Mohammed et al., 2020c (link)).
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