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000291925 1001_ $$00009-0007-4733-893X$$aKipura, Tobias$$b0
000291925 245__ $$aAutomated Liquid Handling Extraction and Rapid Quantification of Underivatized Amino Acids and Tryptophan Metabolites from Human Serum and Plasma Using Dual-Column U(H)PLC-MRM-MS and Its Application to Prostate Cancer Study.
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000291925 520__ $$aAmino acids (AAs) and their metabolites are important building blocks, energy sources, and signaling molecules associated with various pathological phenotypes. The quantification of AA and tryptophan (TRP) metabolites in human serum and plasma is therefore of great diagnostic interest. Therefore, robust, reproducible sample extraction and processing workflows as well as rapid, sensitive absolute quantification are required to identify candidate biomarkers and to improve screening methods. We developed a validated semi-automated robotic liquid extraction and processing workflow and a rapid method for absolute quantification of 20 free, underivatized AAs and six TRP metabolites using dual-column U(H)PLC-MRM-MS. The extraction and sample preparation workflow in a 96-well plate was optimized for robust, reproducible high sample throughput allowing for transfer of samples to the U(H)PLC autosampler directly without additional cleanup steps. The U(H)PLC-MRM-MS method, using a mixed-mode reversed-phase anion exchange column with formic acid and a high-strength silica reversed-phase column with difluoro-acetic acid as mobile phase additive, provided absolute quantification with nanomolar lower limits of quantification within 7.9 min. The semi-automated extraction workflow and dual-column U(H)PLC-MRM-MS method was applied to a human prostate cancer study and was shown to discriminate between treatment regimens and to identify metabolites responsible for discriminating between healthy controls and patients on active surveillance.
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000291925 650_7 $$2Other$$aLC-MS
000291925 650_7 $$2Other$$aamino acids
000291925 650_7 $$2Other$$aautomation
000291925 650_7 $$2Other$$amixed-mode chromatography
000291925 650_7 $$2Other$$aprostate cancer
000291925 650_7 $$2Other$$atryptophan metabolites analysis
000291925 7001_ $$00009-0001-1816-2089$$aHotze, Madlen$$b1
000291925 7001_ $$aHofer, Alexa$$b2
000291925 7001_ $$aEgger, Anna-Sophia$$b3
000291925 7001_ $$00009-0003-3207-0230$$aTimpen, Lea E$$b4
000291925 7001_ $$0P:(DE-He78)14aa02d2ca0515d0c53f1d6678e3ca34$$aOpitz, Christiane$$b5$$udkfz
000291925 7001_ $$00000-0001-8956-9508$$aTownsend, Paul A$$b6
000291925 7001_ $$aGethings, Lee A$$b7
000291925 7001_ $$aThedieck, Kathrin$$b8
000291925 7001_ $$00000-0002-5804-6031$$aKwiatkowski, Marcel$$b9
000291925 773__ $$0PERI:(DE-600)2662251-8$$a10.3390/metabo14070370$$gVol. 14, no. 7, p. 370 -$$n7$$p370$$tMetabolites$$v14$$x2218-1989$$y2024
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