000291925 001__ 291925 000291925 005__ 20241030142645.0 000291925 0247_ $$2doi$$a10.3390/metabo14070370 000291925 0247_ $$2pmid$$apmid:39057693 000291925 0247_ $$2pmc$$apmc:PMC11279291 000291925 0247_ $$2altmetric$$aaltmetric:165860480 000291925 037__ $$aDKFZ-2024-01537 000291925 041__ $$aEnglish 000291925 082__ $$a540 000291925 1001_ $$00009-0007-4733-893X$$aKipura, Tobias$$b0 000291925 245__ $$aAutomated Liquid Handling Extraction and Rapid Quantification of Underivatized Amino Acids and Tryptophan Metabolites from Human Serum and Plasma Using Dual-Column U(H)PLC-MRM-MS and Its Application to Prostate Cancer Study. 000291925 260__ $$aBasel$$bMDPI$$c2024 000291925 3367_ $$2DRIVER$$aarticle 000291925 3367_ $$2DataCite$$aOutput Types/Journal article 000291925 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1722340138_32764 000291925 3367_ $$2BibTeX$$aARTICLE 000291925 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000291925 3367_ $$00$$2EndNote$$aJournal Article 000291925 520__ $$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. 000291925 536__ $$0G:(DE-HGF)POF4-312$$a312 - Funktionelle und strukturelle Genomforschung (POF4-312)$$cPOF4-312$$fPOF IV$$x0 000291925 588__ $$aDataset connected to CrossRef, PubMed, , Journals: inrepo02.dkfz.de 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$$y2024 000291925 8564_ $$uhttps://inrepo02.dkfz.de/record/291925/files/metabolites-14-00370.pdf 000291925 8564_ $$uhttps://inrepo02.dkfz.de/record/291925/files/metabolites-14-00370.pdf?subformat=pdfa$$xpdfa 000291925 909CO $$ooai:inrepo02.dkfz.de:291925$$pVDB 000291925 9101_ $$0I:(DE-588b)2036810-0$$6P:(DE-He78)14aa02d2ca0515d0c53f1d6678e3ca34$$aDeutsches Krebsforschungszentrum$$b5$$kDKFZ 000291925 9131_ $$0G:(DE-HGF)POF4-312$$1G:(DE-HGF)POF4-310$$2G:(DE-HGF)POF4-300$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$aDE-HGF$$bGesundheit$$lKrebsforschung$$vFunktionelle und strukturelle Genomforschung$$x0 000291925 9141_ $$y2024 000291925 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bMETABOLITES : 2022$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0320$$2StatID$$aDBCoverage$$bPubMed Central$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0501$$2StatID$$aDBCoverage$$bDOAJ Seal$$d2023-04-12T15:01:13Z 000291925 915__ $$0StatID:(DE-HGF)0500$$2StatID$$aDBCoverage$$bDOAJ$$d2023-04-12T15:01:13Z 000291925 915__ $$0StatID:(DE-HGF)0030$$2StatID$$aPeer Review$$bDOAJ : Anonymous peer review$$d2023-04-12T15:01:13Z 000291925 915__ $$0LIC:(DE-HGF)CCBYNV$$2V:(DE-HGF)$$aCreative Commons Attribution CC BY (No Version)$$bDOAJ$$d2023-04-12T15:01:13Z 000291925 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0113$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)1190$$2StatID$$aDBCoverage$$bBiological Abstracts$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)9900$$2StatID$$aIF < 5$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0561$$2StatID$$aArticle Processing Charges$$d2023-10-26 000291925 915__ $$0StatID:(DE-HGF)0700$$2StatID$$aFees$$d2023-10-26 000291925 9201_ $$0I:(DE-He78)B350-20160331$$kB350$$lMetabolischer Crosstalk bei Krebserkrankungen$$x0 000291925 9201_ $$0I:(DE-He78)HD01-20160331$$kHD01$$lDKTK HD zentral$$x1 000291925 980__ $$ajournal 000291925 980__ $$aVDB 000291925 980__ $$aI:(DE-He78)B350-20160331 000291925 980__ $$aI:(DE-He78)HD01-20160331 000291925 980__ $$aUNRESTRICTED