Simultaneous monitoring of monoamines, amino acids, nucleotides and neuropeptides by liquid chromatography-tandem mass spectrometry and its application to neurosecretion in bovine chromaffin cells.

摘要: The primary functions of adrenal medullary chromaffin cells are the synthesis and storage in their vesicles catecholamines noradrenaline (NA) adrenaline (AD), subsequent release into bloodstream by Ca2+ -dependent exocytosis under conditions fear or stress (fight flight response). Several monoamines, nucleotides opiates, such as leucine-enkephalin (LENK) methionine-enkephalin (MENK), also co-stored co-released with catecholamines. However, other neurotransmitters have not been studied depth. Here, we present a novel high-resolution liquid chromatography-tandem mass spectrometry approach for simultaneous monitoring 14 compounds stored released bovine (BCCs). We validated analytical method according to recommendations EMA FDA testing matrix effect, selectivity, sensitivity, precision, accuracy, stability carry-over. After on six batches BCCs from different cultures, enabled quantitative determination monoamines (AD, NA, dopamine, serotonin, 5-hydroxyindoleacetic acid, histamine metanephrine), amino acids (L-glutamic γ-aminobutyric acid), (adenosine 5'-diphosphate, adenosine 5'-monophosphate, cyclic 5'-monophosphate) neuropeptides (LENK MENK) intracellular content, basal secretion acetylcholine induced BBCs. used here us determine levels same BCC batch only 16 min. This will make it possible study regulatory mechanisms signaling, endocytosis using neurotrophic factors and/or secretagogues stimuli cultures. Our is actually being applied human plasma samples therapeutic areas where sympathoadrenal axis involved situations Alzheimer's disease, migraine cirrhosis, improve diagnosis clinical practice. Copyright © 2016 John Wiley & Sons, Ltd.