In recent years, use of supercritical-fluid chromatography (SFC) with CO2 as the mobile phase has been expanding in the research laboratory and industry since it is considered to be a green analytical method. This technique offers numerous advantages, such as good separation and sensitive detection, short analysis times, and stability of analytes. In this study, a method for quantification of N-acyl homoserine lactones (AHLs), signaling molecules responsible for cell-to-cell communication initially discovered in bacteria, by SFC coupled with high-resolution mass spectrometry (HRMS) was developed. The SFC conditions and MS ionization settings were optimized to obtain the best separation and greatest sensitivity. The optimal analysis conditions allowed quantification of up to 30 AHLs in a single run within 16 min with excellent linearity (R2 > 0.998) and sensitivity (picogram level). This method was then applied to study AHL production by one Gram-negative endophytic bacterium, Paraburkholderia sp. BSNB-0670. Nineteen known AHLs were detected, and nine abundant HSLs were quantified. To further investigate the production of uncommon AHLs, a molecular networking approach was applied on the basis of the SFC-HRMS/MS data. This led to additional identification of four unknown AHLs annotated as N-3-hydroxydodecanoylol homoserine lactone, N-3-hydroxydodecadienoyl homoserine lactone, and N-3-oxododecenoyl homoserine lactones (two isomers).

中文翻译：

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超临界流体色谱与高分辨率质谱联用对细菌群体感应中的N-酰基高丝氨酸内酯进行注释和定量。

近年来，由于以二氧化碳为流动相的超临界流体色谱法（SFC）被认为是一种绿色分析方法，因此在研究实验室和工业界已在不断扩展。该技术具有许多优点，例如良好的分离和灵敏的检测，较短的分析时间以及分析物的稳定性。在这项研究中，开发了一种通过SFC和高分辨率质谱（HRMS）对N-酰基高丝氨酸内酯（AHL）进行定量的方法，AHL是细菌中最初发现的负责细胞间通信的信号分子。优化了SFC条件和MS电离设置，以获得最佳分离效果和最高灵敏度。最佳分析条件可在16分钟内以优异的线性度（R2> 0）一次定量分析多达30种AHL。998）和灵敏度（皮克级）。然后将该方法用于研究革兰氏阴性内生细菌Paraburkholderia sp。的AHL产生。BSNB-0670。检测到19个已知的AHL，并对9个丰富的HSL进行了量化。为了进一步研究罕见AHL的产生，在SFC-HRMS / MS数据的基础上应用了分子网络方法。这导致另外鉴定了四个未知的AHL，分别标注为N-3-羟基十二烷醇高丝氨酸内酯，N-3-羟基十二碳二烯酰基高丝氨酸内酯和N-3-氧十二碳烯酰基高丝氨酸内酯（两种异构体）。为了进一步研究罕见AHL的产生，在SFC-HRMS / MS数据的基础上应用了分子网络方法。这导致另外鉴定了四个未知的AHL，分别标注为N-3-羟基十二烷醇高丝氨酸内酯，N-3-羟基十二碳二烯酰基高丝氨酸内酯和N-3-氧十二碳烯酰基高丝氨酸内酯（两种异构体）。为了进一步研究罕见AHL的产生，在SFC-HRMS / MS数据的基础上应用了分子网络方法。这导致另外鉴定了四个未知的AHL，分别标注为N-3-羟基十二烷醇高丝氨酸内酯，N-3-羟基十二碳二烯酰基高丝氨酸内酯和N-3-氧十二碳烯酰基高丝氨酸内酯（两种异构体）。