Glucosinolates are biologically active secondary metabolites in Brassicaceae plants that play a critical role in positive and negative interactions between plants and root-associated microbial communities. The aim of this study was to develop a reversed-phase liquid chromatography method to quantify and identify intact glucosinolates in the root of Arabidopsis thaliana (Arabidopsis) grown in non-sterile natural soil by using liquid chromatography-hybrid triple quadruple-linear ion trap (LC-QqQ(LIT)) mass spectrometry. The Synergi Fusion C18-based column was found to be effective for sufficient retention and separation of nine intact glucosinolates without the need for time-consuming desulfation or ion-pairing steps. Method validation results showed satisfactory inter-day and intra-day precision for all glucosinolates except for 4-hydroxyglucobrassicin. Good inter-day and intra-day accuracy and recovery results were observed for glucoiberin, gluconapin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrassicin. However, for 4-hydroxyglucobrassicin, glucoraphanin and glucoerucin corrections to quantification results might be necessary since the recovery and accuracy results were not optimal. Matrix effects were shown to have a negligible effect on the ionization of all target analytes. The established liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was applied to quantify target intact glucosinolates in Arabidopsis root crude extract of four different wild-type accessions where differences in terms of concentration and composition of intact glucosinolates were observed. Employment of sensitive and selective precursor ion survey scan of m/z 97 in combination with the information-dependent acquisition (IDA) of the enhanced product ion (EPI) dependent scan (Prec97-IDA-EPI) using LC-QqQ(LIT) provided high confidence in structural characterization of diverse intact glucosinolate profiles in complex Arabidopsis root crude extract.

中文翻译：

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使用LC-QqQ（LIT）-MS / MS定量和鉴定拟南芥根中完整芥子油苷的分析方法

芥子油苷是十字花科植物中具有生物活性的次生代谢产物，在植物与根相关微生物群落之间的正负相互作用中起关键作用。这项研究的目的是开发一种反相液相色谱法，以定量和鉴定拟南芥（Arabidopsis thaliana（Arabidopsis）根中的完整芥子油苷。液相色谱-三重四重线性离子阱（LC-QqQ（LIT））质谱法在非无菌天然土壤中生长）。发现基于Synergi Fusion C18的色谱柱可有效保留和分离9种完整的芥子油苷，而无需进行费时的脱硫或离子对步骤。方法验证结果表明，除4-羟基葡萄糖基溴青霉素外，所有芥子油苷的日间和日间精度均令人满意。葡糖胺，葡糖那平，葡糖溴素，4-甲氧基葡糖素和新葡糖素的日间和日间准确性和回收率结果均良好。但是，对于4-羟基葡糖嘧啶，由于回收率和准确性结果不是最佳的，可能需要校正葡聚糖，葡糖苷和葡聚糖。基质效应对所有目标分析物的电离影响可忽略不计。建立的液相色谱-串联质谱（LC-MS / MS）方法用于定量目标中完整的芥子油苷四种不同野生型种的拟南芥根粗提物，在其中观察到完整芥子油苷的浓度和组成存在差异。提供了使用m / z 97的敏感和选择性前体离子调查扫描，结合使用LC-QqQ（LIT）的增强型产物离子（EPI）依赖扫描（Prec97-IDA-EPI）的信息依赖采集（IDA）对复杂拟南芥根粗提物中各种完整芥子油苷轮廓的结构表征具有高度信心。