<meta charset="utf8"></head><body>收集 LC 和 TIMS-TOF MS/MS 的正交性以筛选芬太尼类似物

<ol>
	<li>Gladden, R. M., Martinez, P., Seth, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fentanyl+Law+Enforcement+Submissions+and+Increases+in+Synthetic+Opioid-Involved+Overdose+Deaths+-+27+States,+2013-2014.">Fentanyl Law Enforcement Submissions and Increases in Synthetic Opioid-Involved Overdose Deaths - 27 States, 2013-2014.</a> Morbidity and Mortality Weekly Report. 65 (33), 837-843 (2016).</li><li>Rudd, R. A., Seth, P., David, F., Scholl, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increases+in+Drug+and+Opioid-Involved+Overdose+Deaths+-+United+States,+2010-2015.">Increases in Drug and Opioid-Involved Overdose Deaths - United States, 2010-2015.</a> Morbidity and Mortality Weekly Report. 65, 1445-1452 (2016).</li><li>Department of Justice Drug Enforcement Administration. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drugs+of+Abuse:+A+DEA+Resource+Guide.">Drugs of Abuse: A DEA Resource Guide.</a> Department of Justice Drug Enforcement Administration. , (2022).</li><li>Vardanyan, R. S., Hruby, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fentanyl-related+compounds+and+derivatives:+current+status+and+future+prospects+for+pharmaceutical+applications.">Fentanyl-related compounds and derivatives: current status and future prospects for pharmaceutical applications.</a> Future medicinal chemistry. 6 (4), 385-412 (2014).</li><li>Center for Forensic Science Research and Education (CFSRE). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Q4+2022+NPS+Opioids+Trend+Report.">Q4 2022 NPS Opioids Trend Report.</a> Center for Forensic Science Research and Education (CFSRE). , (2022).</li><li>Drug Enforcement Administration. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fentanyl+SIgnature+Profiling+Program+Report.">Fentanyl SIgnature Profiling Program Report.</a> Drug Enforcement Administration. , (2019).</li><li>Drug Enforcement Administration. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+a+Chemical+Precursor+Used+in+the+Illicit+Manufacture+of+Fentanyl+as+a+List+I+Chemical.">Control of a Chemical Precursor Used in the Illicit Manufacture of Fentanyl as a List I Chemical.</a> Drug Enforcement Administration. , (2007).</li><li>Valdez, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gas+Chromatography-Mass+Spectrometry+Analysis+of+Synthetic+Opioids+Belonging+to+the+Fentanyl+Class:+A+Review.">Gas Chromatography-Mass Spectrometry Analysis of Synthetic Opioids Belonging to the Fentanyl Class: A Review.</a> Critical Reviews in Analytical Chemistry. 52 (8), 1938-1968 (2022).</li><li>Churley, M., Cuadra-Rodriguez, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+Note:+Screen+More+Drugs+with+the+Agilent+GC/MS+Toxicology+Analyzer+with+a+High+Efficiency+Source.">Application Note: Screen More Drugs with the Agilent GC/MS Toxicology Analyzer with a High Efficiency Source.</a> Agilent Technologies Incorporation. , (2017).</li><li>Gerace, E., Salomone, A., Vincenti, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analytical+Approaches+in+Fatal+Intoxication+Cases+Involving+New+Synthetic+Opioids.">Analytical Approaches in Fatal Intoxication Cases Involving New Synthetic Opioids.</a> Curr Pharm Biotechnol. 19 (2), 113-123 (2018).</li><li>McIntyre, I. M., Trochta, A., Gary, R. D., Malamatos, M., Lucas, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Acute+Acetyl+Fentanyl+Fatality:+A+Case+Report+With+Postmortem+Concentrations.">An Acute Acetyl Fentanyl Fatality: A Case Report With Postmortem Concentrations.</a> Journal of Analytical Toxicology. 39 (6), 490-494 (2015).</li><li>Marchei, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+synthetic+opioids+in+biological+and+non-biological+matrices:+A+review+of+current+analytical+methods.">New synthetic opioids in biological and non-biological matrices: A review of current analytical methods.</a> Trends in Analytical Chemistry. 102, 1-15 (2018).</li><li>Shoff, E. N., Zaney, M. E., Kahl, J. H., Hime, G. W., Boland, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Qualitative+Identification+of+Fentanyl+Analogs+and+Other+Opioids+in+Postmortem+Cases+by+UHPLC-Ion+Trap-MSn.">Qualitative Identification of Fentanyl Analogs and Other Opioids in Postmortem Cases by UHPLC-Ion Trap-MSn.</a> Journal of Analytical Toxicology. 41 (6), 484-492 (2017).</li><li>Drug Enforcement Administration. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scientific+Working+Group+for+the+Analysis+of+Seized+Drugs+(SWGDRUG)+Recommendations.">Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) Recommendations.</a> Drug Enforcement Administration. , (2022).</li><li>Peters, F. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+comparison+of+four+different+workup+procedures+for+systematic+toxicological+analysis+of+urine+samples+using+gas+chromatography-mass+spectrometry.">A systematic comparison of four different workup procedures for systematic toxicological analysis of urine samples using gas chromatography-mass spectrometry.</a> Analytical and Bioanalytical Chemistry. 393 (2), 735-745 (2009).</li><li>Remane, D., Meyer, M. R., Peters, F. T., Wissenbach, D. K., Maurer, H. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+and+simple+procedure+for+liquid-liquid+extraction+of+136+analytes+from+different+drug+classes+for+development+of+a+liquid+chromatographic-tandem+mass+spectrometric+quantification+method+in+human+blood+plasma.">Fast and simple procedure for liquid-liquid extraction of 136 analytes from different drug classes for development of a liquid chromatographic-tandem mass spectrometric quantification method in human blood plasma.</a> Analytical and Bioanalytical Chemistry. 397 (6), 2303-2314 (2010).</li><li>Cayman Chemical. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supplemental+Material:+Differentiation+of+Isobaric+and+Isomeric+Fentanyl+Analogs+by+Gas+Chromatography-Mass+Spectrometry+(GC-MS).">Supplemental Material: Differentiation of Isobaric and Isomeric Fentanyl Analogs by Gas Chromatography-Mass Spectrometry (GC-MS).</a> Cayman Chemical. , (2018).</li><li>Armenian, P., Vo, K. T., Barr-Walker, J., Lynch, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fentanyl,+fentanyl+analogs+and+novel+synthetic+opioids:+A+comprehensive+review.">Fentanyl, fentanyl analogs and novel synthetic opioids: A comprehensive review.</a> Neuropharmacology. 134, 121-132 (2018).</li><li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+application+of+a+High-Resolution+mass+spectrometry+method+for+the+detection+of+fentanyl+analogs+in+urine+and+serum.">Development and application of a High-Resolution mass spectrometry method for the detection of fentanyl analogs in urine and serum.</a> Journal of Mass Spectrometry and Advances in the Clinical Lab. 26, 1-6 (2022).</li><li>Concheiro, M., Chesser, R., Pardi, J., Cooper, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postmortem+Toxicology+of+New+Synthetic+Opioids.">Postmortem Toxicology of New Synthetic Opioids.</a> Frontiers in Pharmacology. 9, 1210 (2018).</li><li>Mohr, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Novel+Synthetic+Opioids+U-47700,+U-50488+and+Furanyl+Fentanyl+by+LC-MS/MS+in+Postmortem+Casework.">Analysis of Novel Synthetic Opioids U-47700, U-50488 and Furanyl Fentanyl by LC-MS/MS in Postmortem Casework.</a> Journal of Analytical Toxicology. 40 (9), 709-717 (2016).</li><li>Moody, M. T., Diaz, S., Shah, P., Papsun, D., Logan, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+fentanyl+analogs+and+novel+synthetic+opioids+in+blood,+serum/plasma,+and+urine+in+forensic+casework.">Analysis of fentanyl analogs and novel synthetic opioids in blood, serum/plasma, and urine in forensic casework.</a> Drug Testing and Analysis. 10 (9), 1358-1367 (2018).</li><li>Seither, J., Reidy, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confirmation+of+Carfentanil,+U-47700+and+Other+Synthetic+Opioids+in+a+Human+Performance+Case+by+LC-MS-MS.">Confirmation of Carfentanil, U-47700 and Other Synthetic Opioids in a Human Performance Case by LC-MS-MS.</a> Journal of Analytical Toxicology. 41 (6), 493-497 (2017).</li><li>Strayer, K. E., Antonides, H. M., Juhascik, M. P., Daniulaityte, R., Sizemore, I. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LC-MS/MS-Based+Method+for+the+Multiplex+Detection+of+24+Fentanyl+Analogues+and+Metabolites+in+Whole+Blood+at+Sub+ng+mL(-1)+Concentrations.">LC-MS/MS-Based Method for the Multiplex Detection of 24 Fentanyl Analogues and Metabolites in Whole Blood at Sub ng mL(-1) Concentrations.</a> ACS Omega. 3 (-1), 514-523 (2018).</li><li>Kahl, J. H., Gonyea, J., Humphrey, S. M., Hime, G. W., Boland, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Analysis+of+Fentanyl+and+Six+Fentanyl+Analogs+in+Postmortem+Specimens+by+UHPLC-MS-MS.">Quantitative Analysis of Fentanyl and Six Fentanyl Analogs in Postmortem Specimens by UHPLC-MS-MS.</a> Journal of Analytical Toxicology. 42 (8), 570-580 (2018).</li><li>Fogarty, M. F., Papsun, D. M., Logan, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Fentanyl+and+18+Novel+Fentanyl+Analogs+and+Metabolites+by+LC-MS-MS,+and+report+of+Fatalities+Associated+with+Methoxyacetylfentanyl+and+Cyclopropylfentanyl.">Analysis of Fentanyl and 18 Novel Fentanyl Analogs and Metabolites by LC-MS-MS, and report of Fatalities Associated with Methoxyacetylfentanyl and Cyclopropylfentanyl.</a> Journal of Analytical Toxicology. 42 (9), 592-604 (2018).</li><li>Lurie, I. S., Berrier, A. L., Casale, J. F., Iio, R., Bozenko, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profiling+of+illicit+fentanyl+using+UHPLC-MS/MS.">Profiling of illicit fentanyl using UHPLC-MS/MS.</a> Forensic Science International. 220 (1-3), 191-196 (2012).</li><li>Krajewski, L. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+the+fentanyl+analog+screening+kit+toward+the+identification+of+emerging+synthetic+opioids+in+human+plasma+and+urine+by+LC-QTOF.">Application of the fentanyl analog screening kit toward the identification of emerging synthetic opioids in human plasma and urine by LC-QTOF.</a> Toxicology Letters. 320, 87-94 (2020).</li><li>Pirok, B. W. J., Stoll, D. R., Schoenmakers, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+Developments+in+Two-Dimensional+Liquid+Chromatography:+Fundamental+Improvements+for+Practical+Applications.">Recent Developments in Two-Dimensional Liquid Chromatography: Fundamental Improvements for Practical Applications.</a> Analytical Chemistry. 91 (1), 240-263 (2018).</li><li>Keller, T., Keller, A., Tutsch-Bauer, E., Monticelli, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+ion+mobility+spectrometry+in+cases+of+forensic+interest.">Application of ion mobility spectrometry in cases of forensic interest.</a> Forensic Science International. 161 (2-3), 130-140 (2006).</li><li>Klingberg, J., Keen, B., Cawley, A., Pasin, D., Fu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developments+in+high-resolution+mass+spectrometric+analyses+of+new+psychoactive+substances.">Developments in high-resolution mass spectrometric analyses of new psychoactive substances.</a> Archives of Toxicology. 96 (4), 949-967 (2022).</li><li>Fleming, S. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+U-47700,+a+Novel+Synthetic+Opioid,+in+Human+Urine+by+LC-MS-MS+and+LC-QToF.">Analysis of U-47700, a Novel Synthetic Opioid, in Human Urine by LC-MS-MS and LC-QToF.</a> Journal of Analytical Toxicology. 41 (3), 173-180 (2017).</li><li>Guale, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+LC-TOF-MS+screening+for+drugs,+metabolites,+and+collateral+compounds+in+forensic+toxicology+specimens.">Validation of LC-TOF-MS screening for drugs, metabolites, and collateral compounds in forensic toxicology specimens.</a> Journal of Analytical Toxicology. 37 (1), 17-24 (2013).</li><li>Palmquist, K. B., Swortwood, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Data-independent+screening+method+for+14+fentanyl+analogs+in+whole+blood+and+oral+fluid+using+LC-QTOF-MS.">Data-independent screening method for 14 fentanyl analogs in whole blood and oral fluid using LC-QTOF-MS.</a> Forensic Science International. 297, 189-197 (2019).</li><li>Adams, K. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+isomeric+opioids+in+urine+using+LC-TIMS-TOF+MS.">Analysis of isomeric opioids in urine using LC-TIMS-TOF MS.</a> Talanta. 183, 177-183 (2018).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Ammonium formate for HPLC</td>
			<td>Fluka</td>
			<td>17843-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf&#160;Snap-Cap Microcentrifuge Safe-Lock Tubes</td>
			<td>Fisher</td>
			<td>05-402-25</td>
			<td></td>
		</tr>
		<tr>
			<td>ESI-L Low Concentration Tuning mix</td>
			<td>Agilent</td>
			<td>G1969-85000</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl Analog Screening (FAS) Kit&#160;</td>
			<td>Cayman Chemical</td>
			<td>9003237, 9003286, 9003380, 9003381</td>
			<td>kit of 250 snthetic opioids, 210 fentanyl analogs, broken up into one kit and emergent panel versions 1-4</td>
		</tr>
		<tr>
			<td>Formic acid Optima LC/MS</td>
			<td>Fisher</td>
			<td>A117-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Onyx guard column (5 x 4.6 mm)&#160;</td>
			<td>Phenomenex</td>
			<td>CHO-7649</td>
			<td>guard column for C18 columns</td>
		</tr>
		<tr>
			<td>Onyx monolithic C18 HPLC column (100 x 4.6 mm)</td>
			<td>Phenomenex</td>
			<td>CHO-7643</td>
			<td>reverse phase C18 LC column</td>
		</tr>
		<tr>
			<td>Optima grade acetonitrile&#160;</td>
			<td>Fisher</td>
			<td>A996-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Optima grade methanol</td>
			<td>Fisher</td>
			<td>A454-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Optima grade water</td>
			<td>Fisher</td>
			<td>W7-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td>Fisher</td>
			<td>05-719-510</td>
			<td>kit of 1-10 &#181;L, 10-100 &#181;L, and 100-1000 &#181;L pipette</td>
		</tr>
		<tr>
			<td>Pipette tips 10&#181;L</td>
			<td>Fisher</td>
			<td>94060100</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette tips 1000&#181;L</td>
			<td>Fisher</td>
			<td>94056710</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette tips 200&#181;L</td>
			<td>Fisher</td>
			<td>94060310</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate mixer</td>
			<td>IKA&#160;</td>
			<td>MS 3 D S1</td>
			<td>IKA MS 3 digtital</td>
		</tr>
		<tr>
			<td>Prominence LC-20 CE ultrafast liquid chromatograph</td>
			<td>Shimadzu, Japan</td>
			<td></td>
			<td>equiped with DGU-20A5, LC-20AD, SIL-20AC, CTO-20A, SPD-M20A, CBM-20A, SPD-20A</td>
		</tr>
		<tr>
			<td>timsTOF Pro</td>
			<td>Bruker Daltonics Inc., Billerica, MA</td>
			<td></td>
			<td>timsTOF instrument with PASEF&#160;</td>
		</tr>
	</tbody>
</table>

fentanyl analog screening using lc-tims-tof ms/ms

<meta charset="utf8"></head><body>作者聚焦：一种自信地区分和表征芬太尼类似物的有效方法

使用 LC-TIMS-TOF MS/MS 进行芬太尼类似物筛选

The use of fentanyl and the emergence of fentanyl analogs over recent decades has become an increasing concern to the community at large. Fentanyl and its ...

fentanyl-analog-screening-using-lc-tims-tof-msms

Research

Without Section

JoVE Journal

Fentanyl Analog Screening using LC-TIMS-TOF MS/MS

Chemistry

1.6K 次观看.  Florida International University. 我们的研究范围是开发一种区分和表征异构体类似物的方法。本研究旨在研究是否可以开发一种方法来可靠地区分大量，尤其是结构异构体。该协议包含以前未耦合的四种表征模式。对类似物使用四种正交鉴定方法应该可以提高区分当前标准程序难以区分的异构体的能力。芬太尼是阿片类药物流行的一大社会问题。每天都有在非法实验室中发现...

视频: 作者聚焦：一种自信地区分和表征芬太尼类似物的有效方法 

Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications

Free 3-nitrotyrosine (3-NT) has been extensively used as a possible biomarker for oxidative stress. Increased levels of 3-NT have been reported in a wide variety of pathological conditions. However, existing methods lack the sufficient sensitivity and/or specificity necessary to measure the low endogenous level of 3-NT reliably and are too cumbersome for clinical applications. Hence, analytical improvement is urgently needed to accurately quantify the levels of 3-NT and verify the role of 3-NT in pathological conditions. This protocol presents the development of a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) detection combined with a miniaturized solid phase extraction (SPE) for the rapid and accurate measurement of 3-NT in human urine as a non-invasive biomarker for oxidative stress. SPE using a 96-well plate markedly simplified the process by combining sample cleanup and analyte enrichment without tedious derivatization and evaporation steps, reducing solvent consumption, waste disposal, risk of contamination and overall processing time. The employment of 25 mM ammonium acetate (NH4OAc) at pH 9 as the SPE elution solution substantially enhanced the selectivity. Mass spectrometry signal response was improved through adjustment of the multiple reaction monitoring (MRM) transitions. Use of 0.01% HCOOH as additive on a pentafluorophenyl (PFP) column (150 mm x 2.1 mm, 3 &#181;m) improved signal response another 2.5-fold and shortened the overall run time to 7 min. A lower limit of quantitation (LLOQ) of 10 pg/mL (0.044 nM) was achieved, representing a significant sensitivity improvement over the reported assays. This simplified, rapid, selective and sensitive method allows two plates of urine samples (n = 192) to be processed in a 24 h time-period. Considering the markedly improved analytical performance, and non-invasive and inexpensive urine sampling, the proposed assay is beneficial for pre-clinical and clinical studies.

A selective and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method coupled with an efficient solid phase extraction on a mixed-mode cation-exchange (MCX) 96-well microplate was developed for the measurement of free 3-nitrotyrosine (3-NT) in human urine with high throughput, which is suitable for clinical applications.

微型固相萃取和LC-MS / MS检测人尿中3-硝基酪氨酸的临床应用

Free 3-nitrotyrosine (3-NT) has been extensively used as a possible biomarker for oxidative stress. Increased levels of 3-NT have been reported in a wide ...

科研

化学

Natural Product Discovery with LC-MS/MS Diagnostic Fragmentation Filtering: Application for Microcystin Analysis

Natural products are often biosynthesized as mixtures of structurally similar compounds, rather than a single compound. Due to their common structural features, many compounds within the same class undergo similar MS/MS fragmentation and have several identical product ions and/or neutral losses. The purpose of diagnostic fragmentation filtering (DFF) is to efficiently detect all compounds of a given class in a complex extract by screening non-targeted LC-MS/MS datasets for MS/MS spectra that contain class specific product ions and/or neutral losses. This method is based on a DFF module implemented within the open-source MZmine platform that requires sample extracts be analyzed by data-dependent acquisition on a high-resolution mass spectrometer such as quadrupole Orbitrap or quadrupole time-of-flight mass analyzers. The main limitation of this approach is the analyst must first define which product ions and/or neutral losses are specific for the targeted class of natural products. DFF allows for the subsequent discovery of all related natural products within a complex sample, including new compounds. In this work, we demonstrate the effectiveness of DFF by screening extracts of Microcystis aeruginosa, a prominent harmful algal bloom causing cyanobacteria, for the production of microcystins.

Diagnostic fragmentation filtering, implemented into MZmine, is an elegant, post-acquisition approach to screen LC-MS/MS datasets for entire classes of both known and unknown natural products. This tool searches MS/MS spectra for product ions and/or neutral losses that the analyst has defined as being diagnostic for the entire class of compounds.

LC-MCYMS 诊断碎裂过滤的天然产物发现: 微囊藻毒素分析的应用

Natural products are often biosynthesized as mixtures of structurally similar compounds, rather than a single compound. Due to their common structural ...

Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis

This protocol identifies the immunity proteins of the bactericidal enzymes: colicin E3 and bacteriocin, produced by a pathogenic Escherichia coli strain using antibiotic induction, and identified by MALDI-TOF-TOF tandem mass spectrometry and top-down proteomic analysis with software developed in-house. The immunity protein of colicin E3 (Im3) and the immunity protein of bacteriocin (Im-Bac) were identified from prominent b- and/or y-type fragment ions generated by the polypeptide backbone cleavage (PBC) on the C-terminal side of aspartic acid, glutamic acid, and asparagine residues by the aspartic acid effect fragmentation mechanism. The software rapidly scans in silico protein sequences derived from the whole genome sequencing of the bacterial strain. The software also iteratively removes amino acid residues of a protein sequence in the event that the mature protein sequence is truncated. A single protein sequence possessed mass and fragment ions consistent with those detected for each immunity protein. The candidate sequence was then manually inspected to confirm that all detected fragment ions could be assigned. The N-terminal methionine of Im3 was post-translationally removed, whereas Im-Bac had the complete sequence. In addition, we found that only two or three non-complementary fragment ions formed by PBC are necessary to identify the correct protein sequence. Finally, a promoter (SOS box) was identified upstream of the antibacterial and immunity genes in a plasmid genome of the bacterial strain.

Identification of Antibacterial Immunity Proteins in Escherichia coli using MALDI-TOF-TOF-MS/MS and Top-Down Proteomic Analysis

Here we present a protocol for the rapid identification of proteins produced by genomically sequenced pathogenic bacteria using MALDI-TOF-TOF tandem mass spectrometry and top-down proteomic analysis with software developed in-house. Metastable protein ions fragment because of the aspartic acid effect and this specificity is exploited for protein identification.

使用MALDI-TOF-TOF-MS/MS和自上而下的蛋白质组学分析鉴定 大肠杆菌 中的抗菌免疫蛋白

This protocol identifies the immunity proteins of the bactericidal enzymes: colicin E3 and bacteriocin, produced by a pathogenic Escherichia coli strain ...

组蛋白组学分析和翻译后修饰表征

Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry

Histone proteins are highly abundant and conserved among eukaryotes and play a large role in gene regulation as a result of structures known as posttranslational modifications (PTMs). Identifying the position and nature of each PTM or pattern of PTMs in reference to external or genetic factors allows this information to be statistically correlated with biological responses such as DNA transcription, replication, or repair. In the present work, a high-throughput analytical protocol for the detection of histone PTMs from biological samples is described. The use of complementary liquid chromatography, trapped ion mobility spectrometry, and time-of-flight mass spectrometry (LC-TIMS-ToF MS/MS) enables the separation and PTM assignment of the most biologically relevant modifications in a single analysis. The described approach takes advantage of recent developments in dependent data acquisition (DDA) using parallel accumulation in the mobility trap, followed by sequential fragmentation and collision-induced dissociation. Histone PTMs are confidently assigned based on their retention time, mobility, and fragmentation pattern.

作者聚焦： 通过 LC-TIMS-TOF MS/MS 和 PASEF 增强组蛋白 PTM 异构体鉴定

An analytical workflow based on liquid chromatography, trapped ion mobility spectrometry, and time-of-flight mass spectrometry (LC-TIMS-ToF MS/MS) for high confidence and highly reproducible "bottom-up" analysis of histone modifications and identification based on principal parameters (retention time [RT], collision cross section [CCS], and accurate mass-to-charge [m/z] ratio).

使用液相色谱法、捕获离子淌度谱法和飞行时间质谱法进行组蛋白修饰筛选

Histone proteins are highly abundant and conserved among eukaryotes and play a large role in gene regulation as a result of structures known as ...

使用稳定同位素标记筛选脂质结构

Tracing de novo Lipids using Stable Isotope Labeling LC-TIMS-TOF MS/MS

Lipids are highly diverse, and small changes in lipid structures and composition can have profound effects on critical biological functions. Stable isotope labeling (SIL) offers several advantages for the study of lipid distribution, mobilization, and metabolism, as well as de novo lipid synthesis. The successful implementation of the SIL technique requires the removal of interferences from endogenous molecules. In the present work, we describe a high-throughput analytical protocol for the screening of SIL lipids from biological samples; examples will be shown of lipid de novo identification during mosquito ovary development. The use of complementary liquid chromatography trapped ion mobility spectrometry and mass spectrometry allows for the separation and lipids assignment from a single sample in a single scan (&lt;1 h). The described approach takes advantage of recent developments in data-dependent acquisition and data-independent acquisition, using parallel accumulation in the mobility trap followed by sequential fragmentation and collision-induced dissociation. The measurement of SIL at the fatty acid chain level reveals changes in lipid dynamics during the ovary development of mosquitoes. The lipids de novo structures are confidently assigned based on their retention time, mobility, and fragmentation pattern.

作者聚焦：使用稳定同位素标记定量复杂脂质组学样品

Presented here is a method for screening lipid structures through stable isotope labeling by using their retention time, mobility, and fragmentation.

使用稳定同位素标记 LC-TIMS-TOF MS/MS 从 头 追踪脂质

Lipids are highly diverse, and small changes in lipid structures and composition can have profound effects on critical biological functions. Stable ...

Methods Article

使用 LC-TIMS-TOF MS/MS 进行芬太尼类似物筛选