Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University avenue, Toronto, Ontario, M5G 1X5, Canada.
Anal Bioanal Chem. 2012 May;403(3):821-31. doi: 10.1007/s00216-012-5873-2. Epub 2012 Mar 5.
Snow crab is a major fishery in the North Atlantic region. During crab processing the proteins are aerosolized and some are responsible for development of occupational asthma. Tropomyosin and arginine kinase have recently been reported as major snow crab allergens. A liquid chromatographic tandem mass spectrometric method has been developed for simultaneous analysis of these two proteins in air samples collected from processing plants. These proteins were initially isolated then characterized by use of mass spectrometry to determine their primary structure and signature peptides. The signature peptides were chemically synthesized in light and heavy forms and used as standards for developing the multiple-reaction monitoring transitions to monitor allergen levels. A validation study was performed; precision and accuracy were 1.8-8% and 91-104%, respectively. Replicate air samples were collected on air filters from two crab-processing plants in Newfoundland and Labrador (NL) and four located in Quebec. In NL, measured levels of both tropomyosin and arginine kinase were between 1 and 20 ng m(-3). In Quebec plants, however, levels were found to be much higher at 2-2400 ng m(-3). Significant differences were also observed among the plants and individual processing workstations. For the first time arginine kinase has been detected in its aerosolized form in processing plants. In general, levels of the allergens were highest in the butchering and cooking areas; plant design can, however, have a significant effect on levels of the allergens.
雪蟹是北大西洋地区的主要渔业资源。在蟹类加工过程中，蛋白质会被气溶胶化，其中一些会导致职业性哮喘的发生。肌球蛋白轻链和精氨酸激酶最近被报道为雪蟹的主要过敏原。本文建立了一种液相色谱-串联质谱法，用于同时分析来自加工厂的空气样本中这两种蛋白质。这些蛋白质最初通过使用质谱法进行分离和鉴定，以确定它们的一级结构和特征肽。特征肽被化学合成了轻链和重链形式，并用作开发多重反应监测转换以监测过敏原水平的标准。进行了验证研究；精密度和准确度分别为 1.8-8%和 91-104%。在纽芬兰和拉布拉多（NL）的两个蟹类加工厂以及魁北克的四个加工厂的空气过滤器上收集了重复的空气样本。在 NL，肌球蛋白轻链和精氨酸激酶的测量水平在 1-20ng/m³之间。然而，在魁北克的工厂中，水平高达 2-2400ng/m³。还观察到不同工厂和各个加工工作站之间存在显著差异。这是首次在加工工厂中以气溶胶形式检测到精氨酸激酶。一般来说，过敏原水平在屠宰和烹饪区域最高；但是，工厂设计会对过敏原水平产生重大影响。
