Trucksess Mary W, Weaver Carol M, Oles Carolyn J, Rump Lydia V, White Kevin D, Betz Joseph M, Rader Jeanne I
U.S. Food and Drug Administration, 5100 Paint Branch Pkwy, College Park, MD 20740, USA.
J AOAC Int. 2007 Jul-Aug;90(4):1042-9.
Conditions were optimized for the simultaneous, alkaline, aqueous methanol extraction of aflatoxins (AFL), i.e., B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2), and ochratoxin A (OTA) with subsequent purification, isolation, and determination of the toxins in ginseng and ginger. Powdered roots were extracted with methanol-0.5% NaHCO3 solution (7 + 3). After shaking and centrifugation, the supernatant was diluted with 100 mM phosphate buffer containing 1% Tween 20 and filtered through glass microfiber filter paper. The filtrate was then passed through an immunoaffinity column, and the toxins were eluted with methanol. The AFL were separated and determined by reversed-phase liquid chromatography (RPLC) with fluorescence detection after postcolumn UV photochemical derivatization. OTA was separated and determined by RPLC with fluorescence detection. Recoveries of AFL added at 2-16 ng/g and OTA added at 1-8 ng/g to ginseng were 72-80 and 86-95%, respectively. Recoveries of AFL and OTA added to ginger were similar to those for ginseng. A total of 39 commercially available ginger products from 6 manufacturers were analyzed. Twenty-six samples were found to be contaminated with AFL at 1-31 ng/g and 29 samples, with OTA at 1-10 ng/g. Ten samples contained no AFL or OTA. Ten ginseng finished products were also analyzed; 3 contained AFL at 0.1 ng/g and 4 contained OTA at levels ranging from 0.4 to 1.8 ng/g. LC/tandem mass spectrometry with multiple-reaction monitoring of 3 collisionally induced product ions from the protonated molecular ions of OTA, AFB1, and AFG1 was used to confirm the identities of the toxins in extracts of the finished products.
对黄曲霉毒素(AFL),即B1(AFB1)、B2(AFB2)、G1(AFG1)和G2(AFG2)以及赭曲霉毒素A(OTA)的同时碱性水相甲醇提取条件进行了优化,随后对人参和生姜中的毒素进行纯化、分离和测定。将根磨成粉末,用甲醇 - 0.5% NaHCO₃溶液(7 + 3)提取。振荡和离心后,上清液用含1%吐温20的100 mM磷酸盐缓冲液稀释,并用玻璃微纤维滤纸过滤。然后将滤液通过免疫亲和柱,毒素用甲醇洗脱。AFL经柱后紫外光化学衍生后,通过反相液相色谱(RPLC)和荧光检测进行分离和测定。OTA通过RPLC和荧光检测进行分离和测定。向人参中添加2 - 16 ng/g的AFL和1 - 8 ng/g的OTA时,回收率分别为72 - 80%和86 - 95%。添加到生姜中的AFL和OTA的回收率与人参相似。对来自6个制造商的39种市售生姜产品进行了分析。发现26个样品被AFL污染,含量为1 - 31 ng/g,29个样品被OTA污染,含量为1 - 10 ng/g。10个样品未含AFL或OTA。还对10种人参成品进行了分析;3种含有0.1 ng/g的AFL,4种含有含量在0.4至1.8 ng/g之间的OTA。采用液相色谱/串联质谱法,对OTA、AFB1和AFG1的质子化分子离子的3个碰撞诱导产物离子进行多反应监测,以确认成品提取物中毒素的身份。