1290 infinity 2 diode array detector

CoA and CoA-esters were detected with Agilent 1290 Infinity II UHPLC using a reversed-phase C18 column (Agilent InfinityLab Poroshell 120 EC-C18 1.9 μm 2.1 × 50 mm column). The following acetonitrile gradient in 10 mM potassium phosphate buffer (pH 7), with a flow rate of 0.55 ml min^–1, was used: from 2 to 8% at 0–2.66 min; from 8 to 30% at 2.66–3.33 min; from 30 to 2% at 3.33–3.68 min; 2% at 3.68–5 min. Retention times were: 2-methylmalonyl-CoA, 0.9 min; 3-hydroxypropionyl-CoA, 1.4 min; acetyl-CoA, 1.6 min; 3-hydroxybutyryl-CoA, 1.8 min; 3-hydroxy-2-methylpropionyl-CoA, 2.0 min; acryloyl-CoA, 2.2 min; propionyl-CoA, 2.4 min; crotonyl-CoA, 2.6 min; methacrylyl-CoA, 2.9 min. For the conversions with (E)-2-octenoyl-CoA, the following acetonitrile gradient in 10 mM potassium phosphate buffer (pH 6.8), with a flow rate of 0.55 ml min^–1, was used: from 2 to 30% at 0–3.33 min; from 30 to 2% at 3.33–3.68 min; 2% at 3.68–5 min. Retention times were: 3-hydroxyoctanoyl-CoA, 2.3 min; 3-oxo-octanoyl-CoA, 2.4 min; (E)-2-octenoyl-CoA, 3.2 min. Reaction products and standard compounds were detected by UV absorbance at 260 nm with 1290 Infinity II diode array detector (Agilent), and the amount of product was calculated from the relative peak area. The identification of the CoA esters was based on co-chromatography with standards and analysis of the UV spectra of the products.

CoA and CoA-esters were detected with Agilent 1290 Infinity II UHPLC using a reversed-phase C18 column (Agilent InfinityLab Poroshell 120 EC-C18 1.9 μm 2.1 mm × 50 mm column). The following acetonitrile gradient in 10 mM potassium phosphate buffer (pH 7) with a flow rate of 0.55 ml min^–1, was used: from 2 to 8% at 0–2.66 min; from 8 to 30% at 2.66–3.33 min; from 30 to 2% at 3.33–3.68 min; 2% at 3.68–5 min. Retention times were: succinyl-CoA, 0.7 min; CoA, 0.9 min; acetyl-CoA, 1.7 min; propionyl-CoA, 2.4 min; butyryl-CoA, 3.4 min. Reaction products and standard compounds were detected by UV absorbance at 260 nm with a 1290 Infinity II diode array detector (Agilent) and the amount of product was calculated from the relative peak area. The identification of the CoA esters was based on co-chromatography with standards and analysis of the UV spectra of the products.