5 amp

The concentrations of ATP, 5′-AMP and adenosine in culture medium were measured using high-performance liquid chromatography (HPLC). The samples were analysed using a Shimadzu HPLC system. ATP, 5′-AMP and adenosine standards (Solarbio, Beijing, China) were used for optimization. The abundances of ATP and its metabolites were represented by the area of each analyte peak.

1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Ferric-tripyridyltriazine (Fe³⁺-TPTZ) is purchased from Beyotime Biotechnology (Shanghai, China). Chromatographic grade analytical standards asparaginic acid, glutamic acid, asparagine, serine, glutamine, histidine, glycine, threonine, citrulline, arginine, alanine, tyrosine, cystine, valine, methionine, tryptophan, phenylalanine, isoleucine, leucine, lysine, hydroxyproline, proline, 5′-CMP, 5′-UMP, 5′-GMP, 5′-IMP, 5′-XMP, 5′-AMP, trehalose, mannitol, quininic acid, l-malic acid and fumaric acid purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). α-Amylase, α-glucosidase, 4-nitrophenyl α-d-glucopyranoside (PNPG), and soluble starch were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). All other reagents were analytical grade and obtained from Chengdu Kelong Chemical Factory (Chengdu, China).