Van Egmond H P, Jonker K M, Poelman M, Scherpenisse P, Stern A G, Wezenbeek P, Bergwerff A A, Van den Top H J
National Institute for Public Health and the Environment, Laboratory for Food and Residue Analyses, Dutch National Reference Laboratory for Marine Biotoxins, Bilthoven, The Netherlands.
Food Addit Contam. 2004 Apr;21(4):331-40. doi: 10.1080/02652030410001662057.
Paralytic shellfish poisoning toxins are produced by dinoflagellates. Shellfish filtering these unicellular algae will accumulate the toxins and pose a health risk when consumed by man. In the European Union, paralytic shellfish poisoning toxins in bivalve molluscs are regulated at a maximum content of 80 microg/100 g (91/492/EEC). The current reference method in the European Union is the mouse bioassay, but alternative methods including the liquid chromatography methodology are preferred for ethical reasons. Analyses of suspected shellfish batches revealed, however, unacceptable differences in results reported by a small group of Dutch laboratories all using liquid chromatography methods with precolumn derivatization, followed by fluorescence detection. Therefore, a series of proficiency studies were undertaken among these laboratories. In the first three studies, participants were more or less allowed their own choice of method execution details. This approach yielded unsatisfactory results. A fourth study was then initiated in which a standardized method was mandatory. Two types of test material were used in the fourth study: lyophilized Cardium tuberculatum material containing saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX), and lyophilized mussel material containing dc-STX. The latter material was investigated in an interlaboratory study involving 15 participants and was considered as the reference material. Among the four laboratories, coefficients of variation (ANOVA) for C. tuberculatum material were 10% (n = 11) and 9% (n = 12) for STX and dc-STX, respectively, and for the reference material was 8% (n = 12) for dc-STX. The joint efforts showed that variability in analysis results between laboratories that all apply more or less the same method can be drastically improved if the methodology is rigorously standardized.
麻痹性贝类中毒毒素由甲藻产生。滤食这些单细胞藻类的贝类会积累毒素,人类食用后会对健康构成风险。在欧盟,双壳贝类中麻痹性贝类中毒毒素的最高限量为80微克/100克(91/492/EEC)。欧盟目前的参考方法是小鼠生物测定法,但出于伦理原因,包括液相色谱法在内的替代方法更受青睐。然而,对可疑贝类批次的分析显示,一小群荷兰实验室使用柱前衍生化后荧光检测的液相色谱法报告的结果存在不可接受的差异。因此,在这些实验室中进行了一系列能力验证研究。在前三项研究中,参与者在方法执行细节上或多或少可以自行选择。这种方法产生了不尽人意的结果。随后启动了第四项研究,其中强制使用标准化方法。在第四项研究中使用了两种类型的测试材料:含有石房蛤毒素(STX)和脱氨甲酰基石房蛤毒素(dc-STX)的冻干圆蛤材料,以及含有dc-STX的冻干贻贝材料。后一种材料在一项涉及15名参与者的实验室间研究中进行了调查,并被视为参考材料。在这四个实验室中,圆蛤材料中STX和dc-STX的变异系数(方差分析)分别为10%(n = 11)和9%(n = 12),参考材料中dc-STX的变异系数为8%(n = 12)。共同努力表明,如果方法严格标准化,所有或多或少应用相同方法的实验室之间分析结果的变异性可以得到大幅改善。