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一种采用超高效液相色谱法快速测定菌株及商业食品和饲料产品中镰刀菌酸的方法。

A Rapid Method with UPLC for the Determination of Fusaric Acid in Strains and Commercial Food and Feed Products.

作者信息

Chen Zhihong, Luo Qingqing, Wang Mingzi, Chen Bilian

机构信息

College of Life Sciences, Fujian Normal University, 350117 Fuzhou, China.

Engineering Research Center of Industrial Microbiology of Ministry of Education, Fujian Normal University, 350117 Fuzhou, China.

出版信息

Indian J Microbiol. 2017 Mar;57(1):68-74. doi: 10.1007/s12088-016-0617-1. Epub 2016 Aug 23.

Abstract

A rapid, sensitive and validated method for the determination of fusaric acid (FA) in several strains and different commercial food and feed products is reported based on ultra-performance liquid chromatography. This method requires only crude sample by a simple extraction with methanol, and requires a very short time of 8 min for completion. Separation of FA was performed at injection volume of 1 μl with a 20:80 (v/v) water/acetonitrile mobile phase containing 0.1 % formic acid at a flow rate of 0.05 ml/min and detected with UV at 220 nm. Nice linearity and good correlation coefficient (R > 0.99) were obtained in the concentration range of 1-200 μg/ml. Validation was demonstrated using blank samples spiked at three different concentrations with standard solution, and the method yielded more than 98.2 % recovery efficiencies and below 2.56 % R.S.D. when applied in the analysis of FA produced by and a set of transgenic strains of this fungus. Satisfactory recoveries in the range of 79.1-105.8 % and R.S.D lower than 10 % were also obtained for the tested commercial food and feed products. The concentration FA detection in the transgenic strains ranged from 9.65 to 135 μg/kg (0.29-4.05 μg per gram of biomass). However, FA was not detected in most of the commercial products with the exception of niblet, oatmeal, red kidney bean and soybean, for which the concentrations of FA ranged from 2.5 to 18 μg/kg (below the permitted maximum). These results show that the proposed method has a great potential application to analyze FA from different sources rapidly.

摘要

报道了一种基于超高效液相色谱法测定多种菌株以及不同商业食品和饲料产品中镰刀菌酸(FA)的快速、灵敏且经过验证的方法。该方法只需用甲醇简单萃取粗样品,整个分析过程仅需8分钟。FA的分离在进样量为1μl时进行,流动相为含0.1%甲酸的20:80(v/v)水/乙腈,流速为0.05ml/min,于220nm处用紫外检测器检测。在1 - 200μg/ml的浓度范围内获得了良好的线性和较高的相关系数(R > 0.99)。通过向空白样品中添加三种不同浓度的标准溶液进行验证,该方法在分析该真菌及其一组转基因菌株产生的FA时,回收率高于98.2%,相对标准偏差低于2.56%。对于测试的商业食品和饲料产品,回收率在79.1 - 105.8%范围内,相对标准偏差低于10%。转基因菌株中FA的检测浓度范围为9.65至135μg/kg(每克生物量0.29 - 4.05μg)。然而,除了玉米粒、燕麦片、红芸豆和大豆外,大多数商业产品中未检测到FA,这几种产品中FA的浓度范围为2.5至18μg/kg(低于允许的最大值)。这些结果表明,所提出的方法在快速分析不同来源的FA方面具有很大的潜在应用价值。

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