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采用高效液相色谱法和免疫亲和净化法测定玉米片中伏马菌素B1和B2的含量。

Determination of fumonisins B1 and B2 in cornflakes by high performance liquid chromatography and immunoaffinity clean-up.

作者信息

Solfrizzo M, De Girolamo A, Visconti A

机构信息

Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Bari, Italy.

出版信息

Food Addit Contam. 2001 Mar;18(3):227-35. doi: 10.1080/02652030010021963.

Abstract

The determination of fumonisins in cornflakes is a challenging matter as the actually available methods for the analysis of corn do not perform well when applied to this more complex matrix. After testing several factors that may affect the analytical performance, an accurate method for the determination of fumonisin B1 (FB1) and B2 (FB2) in cornflakes has been developed. The method uses immunoaffinity chromatography for clean-up and high performance liquid chromatography (HPLC) for quantification of the toxins. Samples were extracted twice with acetonitrile-methanol-water (25:25:50) and the combined extracts were diluted with phosphate buffered saline (PBS) and applied to a FumoniTest immunoaffinity column. After washing with PBS, fumonisins were eluted from the column with methanol and reacted with o-phthaldialdehyde/2-mercaptoethanol to form fluorescent derivatives. Fumonisin derivatives were analysed by reversed phase HPLC with fluorometric detection using methanol-0.1 M phosphate buffer (77:23; pH adjusted at 3.35) as mobile phase. The average recoveries for FB1 and FB2 spiked in the ranges of 0.33 2.80 microg/g and 0.17-1.40 microg/g were 102.6% and 95.1%, respectively, with average relative standard deviations of 9% and 8%. The limit of quantification for FB1 and FB2 was 0.005 microg/g based on a signal-to-noise ratio of 6.1 by using a sensitive fluorescence detector. The method was used to analyse 18 cornflakes and cornflake cereals samples/for FB1 and FB2 contamination. All but one sample were found to be contaminated, with maximum FB1 and FB2 concentrations of 1.092 microg/g and 0.235 microg/g, respectively. Mean FB1 and FB2 concentrations were 0.157 microg/(g and 0.036 microg/g, respectively.

摘要

由于玉米实际可用的分析方法应用于这种更为复杂的基质时效果不佳,因此测定玉米片中的伏马菌素是一项具有挑战性的工作。在测试了几个可能影响分析性能的因素后,开发出了一种准确测定玉米片中伏马菌素B1(FB1)和B2(FB2)的方法。该方法采用免疫亲和色谱法进行净化,并用高效液相色谱法(HPLC)对毒素进行定量。样品用乙腈 - 甲醇 - 水(25:25:50)提取两次,合并提取物用磷酸盐缓冲盐水(PBS)稀释,然后应用于FumoniTest免疫亲和柱。用PBS洗涤后,伏马菌素用甲醇从柱上洗脱,并与邻苯二甲醛/2 - 巯基乙醇反应形成荧光衍生物。伏马菌素衍生物通过反相HPLC进行分析,采用甲醇 - 0.1M磷酸盐缓冲液(77:23;pH调至3.35)作为流动相进行荧光检测。添加浓度范围为0.33 - 2.80微克/克和0.17 - 1.40微克/克的FB1和FB2的平均回收率分别为102.6%和95.1%,平均相对标准偏差分别为9%和8%。基于使用灵敏荧光检测器时6.1的信噪比,FB1和FB2的定量限为0.005微克/克。该方法用于分析18个玉米片和玉米片谷物样品中的FB1和FB2污染情况。除一个样品外,所有样品均被检测出受到污染,FB1和FB2的最高浓度分别为1.092微克/克和0.235微克/克。FB1和FB2的平均浓度分别为0.157微克/克和0.036微克/克。

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