Oles Carolyn J, Trucksess Mary W
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Office of Regulatory Science, 5100 Paint Branch Pkwy, College Park, MD 20740, USA.
J AOAC Int. 2010 Jul-Aug;93(4):1155-60.
The accuracy, repeatability, and reproducibility characteristics of a published method for measuring levels of fumonisin B1 (FB1) in botanical root products were determined by an AOAC single-laboratory validation procedure. Replicates of 10 test portions of each powdered root product (black cohosh, echinacea, ginger, ginseng, valerian, dong quai, and turmeric) at each spiking level (FB1 at 0, 50, 100, and 200 ng/g) were analyzed on 3 separate days. Test samples were extracted with methanol-acetonitrile-water (25 + 25 + 100, v/v/v). The extracts were centrifuged, the supernatants diluted with phosphate-buffered saline (PBS) containing 1% Tween 20 and filtered, and the filtrates applied to an immunoaffinity column containing antibodies specific for fumonisins. After the column was washed sequentially with PBS and water, the toxin was eluted from the column with 80% methanol, and the eluate dried by lyophilization. The residue was reconstituted with 50% acetonitrile. FB1 was derivatized with a mixture of o-phthaldialdehyde and mercaptoethanol by using an LC autoinjector. Separations were performed with an RP-LC column, and the FB1 derivative was quantified by fluorescence detection. All root products were found to contain FB1 at <10 ng/g. Average within- and between-day recoveries of FB1 from the botanical roots ranged from 67 to 95% and from 68 to 100%, respectively. Total RSD values for within- and between-day repeatability ranged from 5.5 to 26.4%. HorRat values were <1.3 for all of the matrixes examined. The method meets the AOAC method performance criteria at levels of >50 ng/g for the seven botanical roots tested.
采用AOAC单实验室验证程序，测定了一种已发表的用于测量植物根产品中伏马菌素B1（FB1）含量的方法的准确性、重复性和再现性特征。在每个加标水平（FB1分别为0、50、100和200 ng/g）下，对每种粉末状根产品（黑升麻、紫锥菊、生姜、人参、缬草、当归和姜黄）的10个测试部分进行重复测定，并在3个不同日期进行分析。测试样品用甲醇 - 乙腈 - 水（25 + 25 + 100，v/v/v）提取。提取物离心后，上清液用含1%吐温20的磷酸盐缓冲盐水（PBS）稀释并过滤，然后将滤液应用于含有伏马菌素特异性抗体的免疫亲和柱。柱依次用PBS和水洗涤后，用80%甲醇从柱上洗脱毒素，洗脱液经冻干干燥。残留物用50%乙腈复溶。FB1用邻苯二甲醛和巯基乙醇的混合物通过液相色谱自动进样器进行衍生化。使用反相液相色谱柱进行分离，FB1衍生物通过荧光检测进行定量。发现所有根产品中FB1含量均<10 ng/g。从植物根中FB1的日内和日间平均回收率分别为67%至95%和68%至100%。日内和日间重复性的总相对标准偏差（RSD）值范围为5.5%至26.4%。在所检测的所有基质中，HorRat值均<1.3。对于所测试的七种植物根，该方法在>50 ng/g的水平上符合AOAC方法性能标准。
