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Development of an Automated Multi-Injection Shotgun Lipidomics Approach using a Triple Quadrupole Mass Spectrometer



John Bowden, Jackie T. Bangma, John R. Kucklick


Shotgun lipidomics is a well suited approach to monitor lipid alterations due to its ability to scan for varying lipid types on a global, class and individual species level. However, the ability to perform high-throughput shotgun lipidomics has remained challenging due to time-consuming data processing and hardware limitations. To increase the throughput nature of shotgun lipidomics, an automated shotgun lipidomics approach is described utilizing conventional low flow gradient LC analysis (post-injection) coupled with multiple sample injections per sample (on a lipid scan per injection basis). The proposed automated multi-injection approach resulted in a reproducible lipid scanning period of 2.5 min (in a 4.5 min total data acquisition period), thereby providing a sufficient scanning period for performing either mass spectrometric or tandem mass spectrometric analyses. Lipid scanning per injection using the multi-injection shotgun lipidomics approach can be automated using existing LC software and sample queuing/batching. In addition to being simple, robust and reproducible, this approach was also constructed to be cost-effective by using common LC instrumentation, capable of being instrument independent (not proprietary to a specific instrument), and customizable as the data acquisition period can be tailored to perform different scan types, period lengths and scan numbers. Combined with a strategy to create multiple lipid-specific aliquots per sample, the overall approach provides a simple and efficient platform to perform high-throughput lipid profiling.


shotgun lipidomics, lipid profiling, triple quadrupole mass spectrometry, flow-injection analysis, multi-injection shotgun lipidomics, tandem mass spectrometry, direct-infusion


Bowden, J. , Bangma, J. and Kucklick, J. (2015), Development of an Automated Multi-Injection Shotgun Lipidomics Approach using a Triple Quadrupole Mass Spectrometer, Lipids, [online], (Accessed June 24, 2024)


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Created March 12, 2015, Updated January 27, 2020