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Evaluation of Matrix Effects in Quantifying Microbial Secondary Metabolites in Indoor Dust Using Ultraperformance Liquid Chromatograph–Tandem Mass Spectrometer

机译:使用超高效液相色谱-串联质谱仪评估室内灰尘中微生物次生代谢产物的基质效应

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Background Liquid chromatography–tandem mass spectrometry (LC-MSMS) for simultaneous analysis of multiple microbial secondary metabolites (MSMs) is potentially subject to interference by matrix components. Methods We examined potential matrix effects (MEs) in analyses of 31 MSMs using ultraperformance?LC-MSMS. Twenty-one dust aliquots from three buildings (seven aliquots/building) were spiked with seven concentrations of each of the MSMs (6.2 pg/μl–900 pg/μl) and then extracted. Another set of 21 aliquots were first extracted and then, the extract was spiked with the same?concentrations.?We added deepoxy-deoxynivalenol (DOM) to all aliquots as a universal internal standard. Ten microliters of the extract was injected into the ultraperformance LC-MSMS. ME was calculated by subtracting the percentage of the response of analyte in spiked extract to that in neat standard from 100. Spiked extract results were used to create a matrix-matched calibration (MMC) curve for estimating MSM concentration in dust spiked before extraction. Results Analysis of variance was used to examine effects of compound (MSM), building?and concentration on response. MEs (range: 63.4%–99.97%) significantly differed by MSM ( p ??0.01) and building ( p ??0.05). Mean percent recoveries adjusted with DOM and the MMC method were 246.3% (SD?=?226.0) and 86.3% (SD?=?70.7), respectively. Conclusion We found that dust MEs resulted in substantial underestimation in quantifying MSMs and that DOM was not an optimal universal internal standard for the adjustment?but that the MMC method resulted in more accurate and precise recovery compared with DOM. More research on adjustment methods for dust MEs in the simultaneous analyses of multiple MSMs using LC-MSMS is warranted.
机译:背景用于同时分析多种微生物次生代谢物(MSM)的液相色谱-串联质谱(LC-MSMS)可能会受到基质成分的干扰。方法我们使用超高性能LC-MSMS分析了31种MSM,分析了潜在的基质效应(ME)。将来自三个建筑物的二十一个灰尘等分试样(每个建筑物七个等分试样)掺入七种浓度的每种MSM(6.2 pg / μl–900 pg /μl),然后提取。首先提取另一组21个等分试样,然后以相同的浓度加标提取物。我们在所有等分试样中加入了Deepoxy-deoxynivalenol(DOM)作为通用内标。将十微升提取物注入超高性能LC-MSMS中。通过从100中减去加标萃取物中分析物对纯标准品的响应百分比来计算ME。加标萃取物结果用于创建基质匹配校准(MMC)曲线,以估算萃取前加标粉尘中MSM的浓度。结果使用方差分析来检查化合物(MSM),构筑物和浓度对响应的影响。 MEs(范围:63.4%–99.97%)与MSM(p 0.01)和建筑物(p 0.05)有显着差异。用DOM和MMC方法调整后的平均回收率分别为246.3%(SD?=?226.0)和86.3%(SD?=?70.7)。结论我们发现,灰尘MEs在定量MSMs方面被大大低估了,而DOM并不是用于调整的最佳通用内标?但是与DOM相比,MMC方法导致更准确,更准确的回收率。在使用LC-MSMS同时分析多个MSM中,有必要对粉尘ME的调节方法进行更多的研究。

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