A collaborative evaluation of LC-MS/MS based methods for BMAA analysis: soluble bound BMAA found to be an important fraction.

Exposure to β-Ν-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D3BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%-32%), implying that D3BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery ( < 10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis

Comparison of Effects of Endogenous and Exogenous Excitatory Amino Acids on Retzius Nerve Cells of the Leech

In this paper we examine the effects of L-aspartate, L-glutamate, and $-N-oxalylamino-L-alanine (Lathyrus toxin) on Retzius nerve cells of the leech Haemopis sanguisuga. The goal was to compare the electrophysiological effects of endogenous vs. exogenous amino acids, known as potent neurotoxins, through the mechanism of excitotoxicity. We used classical intracellular recordings on Retzius nerve cells in isolated ganglia of the leech, and plotted dose-response curves to compare potencies. Our results show that Lathyrus toxin is more than 200 times more potent in depolarizing the membrane potential on our model than L-aspartate and L-glutamate, which are approximately equipotent