Analysis of β-N-methylamino-L-alanine (BMAA) in spirulina-containing supplements by liquid chromatography-tandem mass spectrometry

Aquatic Biosystems

Pearse McCarron¹  Alan C Logan²  Sabrina D Giddings¹  Michael A Quilliam¹ 

[1] National Research Council Canada, Measurement Science and Standards, Biotoxin Metrology, 1411 Oxford St, Halifax, NS B3H 3Z1, Canada

[2] CAMNR, 23679 Calabasas Road Suite 542, Calabasas, CA 91302, USA

关键词: Food chain;    β-N-methylamino-L-alanine;    BMAA;    Dietary supplements;    Spirulina;   

Others  :  1089038
DOI  :  10.1186/2046-9063-10-5

 received in 2014-07-05, accepted in 2014-08-04,  发布年份 2014

【 摘 要 】

Over the last decade the amino acid beta-N-methylamino-L-alanine (BMAA) has come under intense scrutiny. International laboratory and epidemiological research continues to support the hypothesis that environmental exposure to BMAA (e.g., through dietary practices, water supply) can promote the risk of various neurodegenerative diseases. A wide variety of cyanobacteria spp. have previously been reported to produce BMAA, with production levels dependent upon species, strain and environmental conditions. Since spirulina (Arthrospira spp.) is a member of the cyanobacteria phylum frequently consumed via dietary supplements, the presence of BMAA in such products may have public health implications. In the current work, we have analyzed ten spirulina-containing samples for the presence of BMAA; six pure spirulina samples from two separate raw materials suppliers, and four commercially-available multi-ingredient products containing 1.45 g of spirulina per 8.5 g serving. Because of controversy surrounding the measurement of BMAA, we have used two complementary liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods: one based on reversed phase LC (RPLC) with derivatization and the other based on hydrophilic interaction LC (HILIC). Potential matrix effects were corrected for by internal standardization using a stable isotope labeled BMAA standard. BMAA was not detected at low limits of detection (80 ng/g dry weight) in any of these product samples. Although these results are reassuring, BMAA analyses should be conducted on a wider sample selection and, perhaps, as part of ongoing spirulina production quality control testing and specifications.

【 授权许可】

   
2014 McCarron et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Bradley WG, Borenstein AR, Nelson LM, Codd GA, Rosen BH, Stommel EW, Cox PA: Is exposure to cyanobacteria an environmental risk factor for amyotrophic lateral sclerosis and other neurodegenerative diseases? Amyotroph Lateral Scler Frontotemporal Degener 2013, 14:325-333.
• [2]Chiu AS, Gehringer MM, Welch JH, Neilan BA: Does α-amino-β-methylaminopropionic acid (BMAA) play a role in neurodegeneration? Int J Environ Res Public Health 2011, 8:3728-3746.
• [3]Vega A, Bell EA: α-amino-β-methylaminopropionic acid, a new amino acid from seeds of Cycas circinalis. Phytochem 1967, 6:759-762.
• [4]Spencer PS, Nunn PB, Hugon J, Ludolph AC, Ross SM, Roy DN, Robertson RC: Guam amyotrophic lateral sclerosis-parkinsonism-dementia linked to a plant excitant neurotoxin. Science 1987, 237:517-522.
• [5]Cox PA, Sacks OW: Cycad neurotoxins, consumption of flying foxes, and ALS-PDC disease in Guam. Neurology 2002, 58:956-959.
• [6]Banack SA, Cox PA: Biomagnification of cycad neurotoxins in flying foxes: implications for ALS-PDC in Guam. Neurology 2003, 61:387-389.
• [7]Murch SJ, Cox PA, Banack SA, Steele JC, Sacks OW: Occurrence of beta-methylamino-l-alanine (BMAA) in ALS/PDC patients from Guam. Acta Neurol Scand 2004, 101:267-269.
• [8]Cox PA, Banack SA, Murch SJ: Biomagnification of cyanobacterial neurotoxins and neurodegenerative disease among the Chamorro people of Guam. Proc Natl Acad Sci U S A 2003, 100:13380-13383.
• [9]Cox PA, Banack SA, Murch SJ, Rasmussen U, Tien G, Bidigare RR, Metcalf JS, Morrison LF, Codd GA, Bergman B: Diverse taxa of cyanobacteria produce beta-N-methylamino-L-alanine, a neurotoxic amino acid. Proc Natl Acad Sci U S A 2005, 102:5074-5078.
• [10]Dunlop RA, Cox PA, Banack SA, Rodgers KJ: The non-protein amino acid BMAA is misincorporated into human proteins in place of L-serine causing protein misfolding and aggregation. PLoS One 2013, 8:e75376.
• [11]Rogers KJ: Non-protein amino acids and neurodegeneration: the enemy within. Exp Neurol 2014, 253:192-196.
• [12]Caller TA, Doolin JW, Haney JF, Murby AJ, West KG, Farrar HE, Ball A, Harris BT, Stommel EW: A cluster of amyotrophic lateral sclerosis in New Hampshire: a possible role for toxic cyanobacteria blooms. Amyotroph Lateral Scler 2009, 10(Suppl 2):101-108.
• [13]Masseret E, Banack S, Boumédiène F, Abadie E, Brient L, Pernet F, Juntas-Morales R, Pageot N, Metcalf J, Cox P, Camu W: Dietary BMAA exposure in an amyotrophic lateral sclerosis cluster from southern France. PLoS One 2013, 8:e83406.
• [14]Marles RJ, Barrett ML, Barnes J, Chavez ML, Gardiner P, Ko R, Mahady GB, Low Dog T, Sarma ND, Giancaspro GI, Sharaf M, Griffiths J: United States pharmacopeia safety evaluation of spirulina. Crit Rev Food Sci Nutr 2011, 51:593-604.
• [15]Scott PM, Niedzwiadek B, Rawn DF, Lau BP: Liquid chromatographic determination of the cyanobacterial toxin beta-n-methylamino-L-alanine in algae food supplements, freshwater fish, and bottled water. J Food Prot 2009, 72:1769-1773.
• [16]Spacil Z, Eriksson J, Jonasson S, Rasmussen U, Ilag LL, Bergman B: Analytical protocol for identification of BMAA and DAB in biological samples. Analyst 2010, 135:127-132.
• [17]Banack SA, Metcalf JS, Spacil Z, Downing TG, Downing S, Long A, Nunn PB, Cox PA: Distinguishing the cyanobacterial neurotoxin β-N-methylamino-L-alanine (BMAA) from other diamino acids. Toxicon 2011, 57:730-738.
• [18]Li A, Fan H, Ma F, McCarron P, Thomas K, Tang X, Quilliam MA: Elucidation of matrix effects and performance of solid-phase extraction for LC-MS/MS analysis of β-N-methylamino-L-alanine (BMAA) and 2,4-diaminobutyric acid (DAB) neurotoxins in cyanobacteria. Analyst 2012, 137:1210-1219.
• [19]Jiang L, Aigret B, De Borggraeve WM, Spacil Z, Ilag LL: Selective LC-MS/MS method for the identification of BMAA from its isomers in biological samples. Anal Bioanal Chem 2012, 403:1719-1730.
• [20]Jiang L, Johnston E, Åberg KM, Nilsson U, Ilag LL: Strategy for quantifying trace levels of BMAA in cyanobacteria by LC/MS/MS. Anal Bioanal Chem 2013, 405:1283-1292.
• [21]Glover WB, Liberto CM, McNeil WS, Banack SA, Shipley PR, Murch SJ: Reactivity of β-methylamino-L-alanine in complex sample matrixes complicating detection and quantification by mass spectrometry. Anal Chem 2012, 84:7946-7953.
• [22]Li A, Tian Z, Li J, Yu R, Banack SA, Wang Z: Detection of the neurotoxin BMAA within cyanobacteria isolated from freshwater in China. Toxicon 2010, 55:947-953.
• [23]Faassen EJ: Presence of the neurotoxin BMAA in aquatic ecosystems: what do we really know? Toxins 2014, 6:1109-1138.
• [24]Yoon JY, Park HA, Kang JH, Kim KW, Hur YI, Park JJ, Lee R, Lee HH: Prevalence of dietary supplement use in Korean children and adolescents: insights from Korea National Health and Nutrition Examination Survey 2007–2009. J Korean Med Sci 2012, 27:512-517.
• [25]Kalafati M, Jamurtas AZ, Nikolaidis MG, Paschalis V, Theodorou AA, Sakellariou GK, Koutedakis Y, Kouretas D: Ergogenic and antioxidant effects of spirulina supplementation in humans. Med Sci Sports Exerc 2010, 42:142-151.
• [26]Snyder LR, Cruz-Aguado R, Sadilek M, Galasko D, Shaw CA, Montine TJ: Parkinson-dementia complex and development of a new stable isotope dilution assay for BMAA detection in tissue. Toxicol Appl Pharmacol 2009, 240:180-188.
• [27]Hu Y, Ziffer H: Synthesis and optical resolution of the neurotoxin 2-amino-3-([15N]-methylamino) propanoic acid (BMAA). J Labelled Compounds Radiopharmaceuticals 1990, 28:581-586.
• [28]Yang Y, Park Y, Cassada DA, Snow DD, Rogers DG, Lee J: In vitro and in vivo safety assessment of edible blue-green algae, Nostoc commune var. sphaeroidesKützing and Spirulinaplantensis. Food Chem Toxicol 2011, 49:1560-1564.
• [29]Banji D, Banji OJ, Pratusha NG, Annamalai AR: Investigation on the role of Spirulina platensis in ameliorating behavioural changes, thyroid dysfunction and oxidative stress in offspring of pregnant rats exposed to fluoride. Food Chem 2013, 140:321-331.
• [30]Hwang JH, Chen JC, Chan YC: Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus, expression of NMDA receptor and inflammatory genes. PLoS One 2013, 8:e58215.
• [31]Tobón-Velasco JC, Palafox-Sánchez V, Mendieta L, García E, Santamaría A, Chamorro-Cevallos G, Limón ID: Antioxidant effect of Spirulina (Arthrospira) maxima in a neurotoxic model caused by 6-OHDA in the rat striatum. J Neural Transm 2013, 120:1179-1189.
• [32]Gargouri M, Ghorbel-Koubaa F, Bonenfant-Magné M, Magné C, Dauvergne X, Ksouri R, Krichen Y, Abdelly C, El Feki A: Spirulina or dandelion-enriched diet of mothers alleviates lead-induced damages in brain and cerebellum of newborn rats. Food Chem Toxicol 2012, 50:2303-2310.
• [33]Stromberg I, Gemma C, Vila J, Bickford PC: Blueberry- and spirulina-enriched diets enhance striatal dopamine recovery and induce a rapid, transient microglia activation after injury of the rat nigrostriatal dopamine system. Exp Neurol 2005, 196:298-307.
• [34]Escalona-Cardoso GN, Paniagua-Castro N, Pérez-Pastén R, Chamorro-Cevallos G: Spirulina (arthrospira) protects against valproic acid-induced neural tube defects in mice. J Med Food 2012, 15:1103-1108.
• [35]Pabon MM, Jernberg JN, Morganti J, Contreras J, Hudson CE, Klein RL, Bickford PC: A spirulina-enhanced diet provides neuroprotection in an α-synuclein model of Parkinson’s disease. PLoS One 2012, 7:e45256.
• [36]Abdel-Daim MM, Abuzead SM, Halawa SM: Protective role of Spirulina platensis against acute deltamethrin-induced toxicity in rats. PLoS One 2013, 8:e72991.
• [37]Downing S, Banack SA, Metcalf JS, Cox PA, Downing TG: Nitrogen starvation of cyanobacteria results in the production of β-N-methylamino-L-alanine. Toxicon 2011, 58:187-194.
• [38]Holman BW, Malau-Aduli AE: Spirulina as a livestock supplement and animal feed. J Anim Physiol Anim Nutr 2013, 97:615-623.