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A two-step gas chromatography-tandem mass spectrometry method for measurement of multiple environmental pollutants in human plasma

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Individuals are exposed to a wide variety of chemicals over their lifetime, yet current understanding of mixture toxicology is still limited. We present a two-step analytical method using a gas chromatograph-triple quadrupole mass spectrometer that requires less than 1 mL of sample. The method is applied to 183 plasma samples from a study population of children with autism spectrum disorder, their parents, and unrelated neurotypical children. We selected 156 environmental chemical compounds and ruled out chemicals with detection rates less than 20% of our study cohort (n = 61), as well as ones not amenable to the selected extraction and analytical methods (n = 34). The targeted method then focused on remaining chemicals (n = 61) plus 8 additional polychlorinated biphenyls (PCBs). Persistent pollutants, such as p,p′-dichlorodiphenyldichloroethylene (p,p′-DDE) and PCB congeners 118 and 180, were detected at high frequencies and several previously unreported chemicals, including 2,4,6-trichlorophenol, isosafrole, and hexachlorobutadiene, were frequently detected in our study cohort. This work highlights the benefits of employing a multi-step analytical method in exposure studies and demonstrates the efficacy of such methods for reporting novel information on previously unstudied pollutant exposures.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.


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The authors would like to thank Carles Hernandez-Ferrer for the help with quality control methods and Yufei Lin for the assistance with obtaining the demographic data and advice on data analysis. Thank you also to Bonnie Marion for the help with sample extraction method, to Anthony Macherone and Agilent Technologies for advice and support regarding GC-MS methods and providing their pesticide screening MRM database, and to Kelly Cara and Mei Chung for obtaining the age-subsetted NHANES data.


This research is funded by the US National Institutes of Mental Health under Grant number NIMH R01MH107205.

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Authors and Affiliations



CLJ analyzed extracted plasma samples, extracted mass spectral data, performed statistical analysis, and prepared the initial manuscript. EJ performed sample extraction and assisted with sample analysis. SWK obtained plasma samples and contributed to the study design and manuscript preparation. KDP assisted with the development of analytical methods, managed the research project, and was a major contributor to manuscript preparation. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Sek Won Kong or Kurt D. Pennell.

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Competing interests

The authors declare that they have no competing interests.

Ethics approval and consent to participate

The Institutional Review Board (IRB) of Tufts University determined that study #1712011 was excluded from IRB review and approval.

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Not applicable.

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Responsible Editor: Ester Heath

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Electronic supplementary material

Online Resource 1

MRM Transitions and Collision Energies (CE, in volts) for all chemicals and two internal recovery surrogates measured in human plasma. (ESM_1.docx) (DOCX 85 kb)

Online Resource 2

Instrumental detection limits (IDL) for 31 chemicals. The IDL was calculated using 10 replicate injections of a low-concentration standard in hexane, and therefore does not account for matrix effects or variation introduced from the extraction method. (DOCX 36 kb)

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Johnson, C.L., Jazan, E., Kong, S.W. et al. A two-step gas chromatography-tandem mass spectrometry method for measurement of multiple environmental pollutants in human plasma. Environ Sci Pollut Res 28, 3266–3279 (2021).

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