(1340-A) Method Updates in the Measurement of Heterocyclic Aromatic Amines in Human Urine using Hamilton Starlet Automated Liquid Handling Platform and LC-ESI-MS/MS
Monday, February 5, 2024
12:00 PM – 1:00 PM EST
Location: Exhibit Halls AB
Abstract: Method Updates in the Measurement of Heterocyclic Aromatic Amines in Human Urine using Hamilton Starlet Automated Liquid Handling Platform and LC-ESI-MS/MS
Megan Smith1,2, Sujeewa Piyankarage1, Shrila Mazumder1, and Lanqing Wang1
1 Tobacco and Volatiles Branch, Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, Georgia 2 Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, Tennessee
* The findings and conclusions in this study are those of the authors and do not necessarily represent the views of the U.S. Department of Health and Human Services, or the U.S. Centers for Disease Control and Prevention. Use of trade names and commercial sources is for identification only and does not constitute endorsement by the U.S. Department of Health and Human Services, or the U.S. Centers for Disease Control and Prevention.
Keywords: heterocyclic aromatic amines, human urine, automation, sample data flow, liquid chromatography, tandem mass spectrometry
Word Count: Abstract = 253
Abstract Heterocyclic aromatic amines (HCAAs), many of which are established by the International Agency for Research on Cancer as probable/possible human carcinogens (group 2A/2B), are formed during smoking of cigarettes and cooking of meats at high temperature. Exposure to HCAAs such as 2-amino-9H-pyrido[2,3-b]indole (AAC), 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MAAC), 3-amino-1, 4-dimethyl-5H-pyrido[4,3-b ]indole (TRP1), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (TRP2), 2-amino-6-methyldipyrido[1,2-a:3’,2’-D]imidazole (GLP1), 2-aminodipyrido[1,2-a:3’,2-D]imidazole (GLP2), 2-amino-3-methyl-3H-imidazo[4,5-f]quinolone (IQU), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PHIP), harman (HMU) and norharman (NHM) can be assessed by measuring their concentrations in urine. AAC, MAAC, TRP1, TRP2, GLP1, GLP2, IQU, PHIP, HMN and NHM are measured by isotope-dilution high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (ID-HPLC-ESI-MS/MS). Updates to an existing automated sample preparation method were made to more efficiently quantify these HCAAs, in which, sample aliquot, clean-up and reconstitution are all completed on a Hamilton STARLET™ workstation. This new automated method increased sample throughput by reducing sample clean-up time from 6.5 hours to 5 hours while maintaining precision (inter-run CV < 9%) and accuracy (±10%). This updated method also improves data review throughput by using Indigo ASCENTTM program (Indigo BioAutomation, Inc., version 3.7) for peak integration, quantification, sample QA rule evaluations, and result report file generation. Furthermore, a streamlined sample data flow was created in parallel to the automated method, in which, sample results can be tracked from initial analysis, repeat analysis, and final reporting to laboratory information management system with minimal human intervention. This new automated method and sample data flow are currently applied in small studies that aim to biomonitor “total” (free + conjugated) HCAAs in 24-hour urine samples.