A Collaborative Evaluation of LC-MS/MS Based Methods for BMAA Analysis: Soluble Bound BMAA Found to Be an Important Fraction

Exposure to β-N-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.     

Composition and biological activity of the essential oil from Thapsia minor, a new source of geranyl acetate

The essential oils from aerial parts of Thapsia minor Hoffmanns. & Link (Apiaceae), obtained by hydrodistillation, were analyzed by gas-chromatography and gas-chromatography-mass spectrometry. Oxygenated monoterpenes represented the main fraction with geranyl acetate (82.3-83.0%) as the major constituent, making this plant a good source of this compound.Biological activity of the essential oil was evaluated against several fungal strains. The highest activity was found against dermatophyte strains, Crytococcus neoformans and Candida guillermondii with minimal inhibitory concentrations values from 0.32 μl/ml to 1.25 μl/ml. The antifungal activity could be associated with the significant contribution of the oxygenated monoterpene geranyl acetate.Furthermore, this essential oil demonstrated to have potent anti-inflammatory in vitro activity, at concentrations that did not affect macrophages and keratinocytes viability, being the anti-inflammatory properties of the oil mainly due to geranyl acetate.These favorable results emphasize the antifungal and anti-inflammatory abilities of the molecule geranyl acetate and further support the exploitation of T. minor oil for pharmaceutical industry, in order to find new phytopharmaceuticals.