建立一种用于定量测定玉米片中伏马菌素B1、B2和B3的液相色谱/串联质谱法。

Development of a liquid chromatography/tandem mass spectrometry method for the quantification of fumonisin B1, B2 and B3 in cornflakes.

作者信息

Paepens C, De Saeger S, Van Poucke C, Dumoulin F, Van Calenbergh S, Van Peteghem C

机构信息

Laboratory of Food Analysis, Faculty of Pharmaceutical Sciences, Ghent University, UGent, Harelbekestraat 72, 9000 Ghent, Belgium.

出版信息

Rapid Commun Mass Spectrom. 2005;19(14):2021-9. doi: 10.1002/rcm.2022.

DOI:10.1002/rcm.2022

PMID:15973649

Abstract

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of fumonisin B1 (FB1), B2 (FB2) and B3 (FB3) in cornflakes is described. During method development, special attention was paid to the selection of a suitable internal standard (IS) in order to offer a good alternative for deuterated FB1. In this respect, the C12-sphinganine analogue (2S,3R)-2-aminododecane-1,3-diol was chosen because of its structural similarity to the fumonisin backbone and its chromatographic elution between the target analytes. For the extraction of the fumonisins from the cornflakes matrix, MeOH/H2O (adjusted to pH 4 with 0.1 M HCl; 70:30, v/v), ACN/MeOH/H(2)O (25:25:50, v/v/v) and acidified ACN/MeOH/H2O (25:25:50, v/v/v; pH 4) were evaluated. Preference was given to acidified MeOH/H2O (70:30, v/v) with mean recoveries (n=12) for FB1, FB2 and FB3 of, respectively, 84+/-10, 78+/-7 and 85+/-9%. Cleanup was performed using immunoaffinity columns (FumoniTest, VICAM). The chromatography was performed under isocratic conditions at a flow of 0.3 mL min-1 with a mobile phase consisting of ACN/H2O (60:40, v/v) containing 0.3% formic acid. The mass spectrometer was operated in the positive electrospray ionization (ESI+) mode using multiple reaction monitoring (MRM). An intralaboratory validation was conducted with fortified samples determining limits of detection (LOD), limits of quantification (LOQ), precision, trueness, specificity and measurement uncertainty. The LOD concentrations for FB1, FB2 and FB3 were 20, 7.5 and 12.5 microg/kg. The LOQs were 40 microg/kg for FB1, 15 microg/kg for FB2 and 25 microg/kg for FB3. The coefficients of variation (CVs) under repeatability conditions varied from 11 to 13% for FB1, from 9 to 14% for FB2 and from 7 to 10% for FB3. Under within-laboratory reproducibility conditions, the CVs ranged from 12 to 17% for FB1, from 9 to 16% for FB2 and from 7 to 13% for FB3. The percent bias for FB1 varied from -12 to -10%, while for FB2 and FB3 bias ranged, respectively, from -4 to -2% and from -12 to -5%. The expanded measurement uncertainties for FB1, FB2 and FB3 were, respectively, 19, 18 and 22%.

摘要

描述了一种用于测定玉米片中伏马毒素B1（FB1）、B2（FB2）和B3（FB3）的液相色谱/串联质谱（LC/MS/MS）方法。在方法开发过程中，特别关注了合适内标（IS）的选择，以便为氘代FB1提供一个良好的替代物。在这方面，选择了C12 - 鞘氨醇类似物（2S,3R）-2 - 氨基十二烷 - 1,3 - 二醇，因为它与伏马毒素主链结构相似，且在目标分析物之间具有色谱洗脱特性。对于从玉米片基质中提取伏马毒素，评估了甲醇/水（用0.1 M盐酸调至pH 4；70:30，v/v）、乙腈/甲醇/水（25:25:50，v/v/v）和酸化乙腈/甲醇/水（25:25:50，v/v/v；pH 4）。优先选择酸化甲醇/水（70:30，v/v），FB1、FB2和FB3的平均回收率（n = 12）分别为84±10%、78±7%和85±9%。使用免疫亲和柱（FumoniTest，VICAM）进行净化。色谱分析在等度条件下进行，流速为0.3 mL/min，流动相由含0.3%甲酸的乙腈/水（60:40，v/v）组成。质谱仪在正电喷雾电离（ESI +）模式下使用多反应监测（MRM）运行。对加标样品进行了实验室内部验证，确定了检测限（LOD）、定量限（LOQ）、精密度、准确性、特异性和测量不确定度。FB1、FB2和FB3的LOD浓度分别为20、7.5和12.5 μg/kg。FB1的LOQ为40 μg/kg，FB2为15 μg/kg，FB3为25 μg/kg。在重复性条件下，FB1的变异系数（CV）为11%至13%，FB2为9%至14%，FB3为7%至10%。在实验室内部再现性条件下，FB1的CV为12%至17%，FB2为9%至16%，FB3为7%至13%。FB1的偏差百分比在 - 12%至 - 10%之间，而FB2和FB3的偏差分别在 - 4%至 - 2%和 - 12%至 - 5%之间。FB1、FB2和FB3的扩展测量不确定度分别为19%、18%和22%。