Van de Riet Jeffrey M, Gibbs Ryan S, Chou Faith W, Muggah Patricia M, Rourke Wade A, Burns Garth, Thomas Krista, Quilliam Michael A
Canadian Food Inspection Agency, Dartmouth Laboratory, 1992 Agency Dr, Dartmouth, NS, Canada B3B 1Y9.
J AOAC Int. 2009 Nov-Dec;92(6):1690-704.
A single-laboratory validation study was conducted for the LC post-column oxidation analysis of paralytic shellfish toxins (PST): saxitoxin (STX); neosaxitoxin (NEO); gonyautoxins (GTX) 1-5; decarbamoyl gonyautoxins (dcGTX) 2 and 3; decarbamoyl saxitoxin (dcSTX); and N-sulfocarbamoyl-gonyautoxin-2 and 3 (C1 and C2) in mussels (Mytilus edulis), soft shell clams (Mya arenaria), scallops (Placopectin magellanicus), and oysters (Crassostrea virginicus). The instrumental technique was developed for the analysis of PST in shellfish as an alternative to the precolumn oxidation method, AOAC Official Method 2005.06, and a replacement for the current AOAC biological method 959.08. The method used reversed-phase LC with post-column oxidation and fluorescence detection. Test materials for method recovery were prepared by fortification of blank material with a cocktail of PST. Materials used to determine method repeatability and intermediate precision were prepared by blending blank material with naturally incurred material. The target total toxicity levels evaluated in the study were 0.40, 0.80, and 1.60 mg STX x diHCl equivalents per kilogram [(eq/kg) 1%, 1, and 2 times the regulatory limit]. Linearity, recovery, and within-laboratory precision parameters of the method were evaluated. Correlation coefficients of the calibration curves for all toxins studied were > 0.99. Total toxin recovery ranged from 94 to 106% at the three levels of interest. Repeatability and intermediate precision RSD ranged from 2 to 7% and 2 to 8%, respectively. The method LOD and LOQ (assuming the presence of all toxins) were determined to be equivalent to 0.18 and 0.39 mg STX x diHCl eq/kg. The method is intended for a regulatory framework and will be submitted for an AOAC collaborative study.
针对麻痹性贝类毒素（PST）的液相色谱柱后氧化分析开展了一项单实验室验证研究，分析对象包括贻贝（紫贻贝）、软壳蛤（砂海螂）、扇贝（麦哲伦扁贝）和牡蛎（弗吉尼亚牡蛎）中的石房蛤毒素（STX）、新石房蛤毒素（NEO）、膝沟藻毒素（GTX）1 - 5、脱氨甲酰基膝沟藻毒素（dcGTX）2和3、脱氨甲酰基石房蛤毒素（dcSTX）以及N - 磺基甲酰基 - 膝沟藻毒素 - 2和3（C1和C2）。该仪器技术是为分析贝类中的PST而开发的，可替代柱前氧化法（AOAC官方方法2005.06），并取代现行的AOAC生物学方法959.08。该方法采用反相液相色谱结合柱后氧化和荧光检测。方法回收率的测试材料通过向空白材料中添加PST混合物来制备。用于确定方法重复性和中间精密度的材料通过将空白材料与天然含毒材料混合制备。研究中评估的目标总毒性水平为每千克0.40、0.80和1.60毫克石房蛤毒素二盐酸盐当量[（eq/kg），分别为监管限值的1%、1倍和2倍]。对该方法的线性、回收率和实验室内部精密度参数进行了评估。所研究的所有毒素校准曲线的相关系数均>0.99。在三个目标水平下，总毒素回收率在94%至106%之间。重复性和中间精密度的相对标准偏差（RSD）分别在2%至7%和2%至8%之间。该方法的检测限（LOD）和定量限（LOQ）（假设所有毒素均存在）经测定分别相当于0.18和0.39毫克石房蛤毒素二盐酸盐当量/千克。该方法适用于监管框架，并将提交AOAC协作研究。
