Determination of acetyl coenzyme A in grape berries by high performance liquid chromatography-tandem mass spectrometry

Abstract

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS) method was developed for the determination of acetyl coenzyme A in grape berries using .-propionyl coenzyme A as an internal standard (IS). The sample was extracted with water and then centrifuged for 5 min at 10 000 r/min on a centrifuge at 4 °C, and cLned-up with a C18 solid phase extraction cartridge. The identification and quantification were carried out by using electrospray ionization in positive ion mode (ESI +) with multiple reaction monitoring (MEM). The total run time was 1 min and the elution of both acetyl coenzyme A and.-propio-nyl coenzyme A occurred at about 0. 45 min. This was achieved with a mobile phase consisting of 5 mmol/L ammonium formate-acetonitrile (20: m, v/v) at a flow rate of 0. 4 mL/min on a C18 column. A linear response function was established for the concentration range of 1-2 000 μg/L for acetyl coenzyme A and the correlation coefficient was more than 0. 99. The limit of detection of acetyl coenzyme A was 0. 1 μg/L. The spiked recoveries at 50, 500, 1 000 μg/L ranged from 82. 87% -89. 67% with the relative standard deviations less than 10%. This method is simple, rapid, sensitive and can significantly reduce the loss of acetyl coenzyme A during the experiment. It is suitable for the determination of acetyl coenzyme A in grape berries.