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同位素稀释–超高效液相色谱–串联质谱法测定板栗中5种交链孢霉菌素

Isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry for determination of five alternariol mycotoxins in chestnut

  • 摘要:
    目的  建立同时测定板栗中5种交链孢霉菌素含量的同位素稀释–超高效液相色谱–串联质谱法。
    方法  通过参数优化确定5种目标物的最优质谱条件,分析目标化合物色谱峰的出峰情况,选择最佳的色谱柱和流动相,通过回收率比较不同提取试剂的提取效果以确定最佳的提取试剂,最后对建立的方法进行方法学验证。
    结果  该方法最终使用乙腈–水–甲酸(70+29+1)作为提取试剂对样品前处理。采用BEH C18为色谱柱,流动相为乙腈碳酸氢铵–0.15 mmol/L,方法的基质效应不能忽略,应采用内标法定量分析,5种交链孢霉菌素的检出限为0.10~0.50 μg/kg,定量限为0.30~1.50 μg/kg,回收率80.1%~111.7%,相对标准偏差2.3%~8.2%,相关系数均≥0.999 0 。
    结论  本方法操作简便、快速准确,线性关系、精密度和回收率等均满足方法学要求,适用于板栗中5种交链孢霉菌素含量的同时测定。

     

    Abstract:
    Objective Establishment of an isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (ID-UPLC-MS/MS) method for the simultaneous determination of five alternariol mycotoxins in chestnuts.
    Methods The optimal mass spectrometry conditions for the five target compounds were determined through parameter optimization. The chromatographic peak elution of the target compounds was analyzed, and the optimal chromatographic column and mobile phase were selected. The extraction efficiency of different extraction reagents was compared by recovery experiments to determine the optimal extraction reagent. Finally, the established method was validated.
    Results The method ultimately used acetonitrile-water-formic acid (70 + 29 + 1) as the extraction reagent for sample pretreatment. A BEH C18 column was used with a mobile phase of acetonitrile -0.15 mmol/L ammonium bicarbonate. The matrix effect of the method could not be ignored, and the internal standard method should be used for quantitative analysis. The limits of detection (LODs) for the five alternariol mycotoxins ranged from 0.10 to 0.50 μg/kg, and the limits of quantitation (LOQs) ranged from 0.30 to 1.50 μg/kg. The recoveries ranged from 80.1% to 111.7%, with relative standard deviations (RSDs) ranging from 2.3% to 8.2%. The correlation coefficients were all ≥ 0.999 0.
    Conclusions This method is simple, rapid, and accurate. The linearity, precision, and recovery meet the methodological requirements, and it is suitable for the simultaneous determination of five alternariol mycotoxins in chestnuts.

     

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