Lesiak Ashton D, Fowble Kristen L, Musah Rabi A
Department of Chemistry, University at Albany-SUNY, Albany, NY, USA.
Department of Chemistry, State University of New York at Albany, Albany, NY, USA.
Methods Mol Biol. 2018;1810:207-215. doi: 10.1007/978-1-4939-8579-1_18.
The utility of direct analysis in real time-high resolution mass spectrometry (DART-HRMS) for quantification of a variety of compounds has been explored, but the number of reports of validated methods using this technique is limited. Furthermore, despite the increasing use in crime labs of DART-HRMS for the detection and identification of drugs of abuse, very few published reports have appeared describing how the method can be exploited for the analysis of small molecules of interest within complex matrices such as plant tissues. Herein we describe the steps to be taken to establish a validated quantification method for psychoactive compounds within complex plant matrices through its application to the detection and quantification of atropine in Datura stramonium seeds. Six calibration standard series are analyzed eight times over a period of several days to create a calibration curve. The resulting calibration curve is tested using six quality control samples and finally utilized to determine the concentration of atropine in a D. stramonium seed extract. The linear range for quantification of atropine in this study was found to be comparable to that reported previously using GC, LC, HPLC, and UHPLC-MS methods. Furthermore, the method can be applied to the quantification of other biomarkers in plant materials, despite the complexity of the plant matrix. The speed of the analysis (<10 min for duplicate analysis of 20 samples) and the ability to integrate peaks using accurate masses for specificity are advantages of the DART-HRMS quantification approach.
直接实时分析-高分辨率质谱法(DART-HRMS)用于多种化合物定量分析的实用性已得到探索,但使用该技术的经过验证的方法的报告数量有限。此外,尽管DART-HRMS在犯罪实验室中越来越多地用于检测和鉴定滥用药物,但很少有已发表的报告描述如何利用该方法分析复杂基质(如植物组织)中感兴趣的小分子。在此,我们描述了通过将其应用于曼陀罗种子中阿托品的检测和定量,建立复杂植物基质中精神活性化合物经过验证的定量方法所需采取的步骤。在几天的时间内对六个校准标准系列进行八次分析,以创建校准曲线。使用六个质量控制样品对所得校准曲线进行测试,最后用于测定曼陀罗种子提取物中阿托品的浓度。本研究中阿托品定量的线性范围与先前使用气相色谱(GC)、液相色谱(LC)、高效液相色谱(HPLC)和超高效液相色谱-质谱(UHPLC-MS)方法报道的范围相当。此外,尽管植物基质复杂,但该方法可应用于植物材料中其他生物标志物的定量分析。分析速度(对20个样品进行重复分析<10分钟)以及使用精确质量积分峰以提高特异性的能力是DART-HRMS定量方法的优势。