Shah Syed Muhammad Zaki, Ali Arslan, Khan Muhammad Noman, Khadim Adeeba, Asmari Mufarreh, Uddin Jalal, Musharraf Syed Ghulam
International Center for Chemical and Biological Sciences, H.E.J. Research Institute of Chemistry, University of Karachi, Karachi 75270, Pakistan.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.
Pharmaceuticals (Basel). 2022 Jul 21;15(7):901. doi: 10.3390/ph15070901.
Data-independent acquisition (DIA) based strategies have been explored in recent years for improving quantitative analysis of metabolites. However, the data analysis is challenging for DIA methods as the resulting spectra are highly multiplexed. Thus, the DIA mode requires advanced software analysis to facilitate the data deconvolution process. We proposed a pipeline for quantitative profiling of pharmaceutical drugs and serum metabolites in DIA mode after comparing the results obtained from full-scan, Data-dependent acquisition (DDA) and DIA modes. using open-access software. Pharmaceutical drugs (10) were pooled in healthy human serum and analysed by LC-ESI-QTOF-MS. MS1 full-scan and Data-dependent (MS2) results were used for identification using MS-DIAL software while deconvolution of MS1/MS2 spectra in DIA mode was achieved by using Skyline software. The results of acquisition methods for quantitative analysis validated the remarkable analytical performance of the constructed workflow, proving it to be a sensitive and reproducible pipeline for biological complex fluids.
近年来，基于数据非依赖采集（DIA）的策略已被用于改善代谢物的定量分析。然而，由于DIA方法产生的光谱具有高度多重性，其数据分析具有挑战性。因此，DIA模式需要先进的软件分析来促进数据去卷积过程。在比较全扫描、数据依赖采集（DDA）和DIA模式获得的结果后，我们提出了一种用于在DIA模式下对药物和血清代谢物进行定量分析的流程，该流程使用开源软件。将10种药物混合在健康人血清中，通过液相色谱-电喷雾-四极杆飞行时间质谱（LC-ESI-QTOF-MS）进行分析。MS1全扫描和数据依赖（MS2）结果使用MS-DIAL软件进行鉴定，而DIA模式下MS1/MS2光谱的去卷积则通过Skyline软件实现。定量分析采集方法的结果验证了所构建工作流程卓越的分析性能，并证明其是一种用于生物复杂流体的灵敏且可重复的流程。
