Gas chromatography negative chemical ionization mass spectrometry (GC-NCI-MS) is a preferred instrumental approach for the trace and ultra-trace analysis of various toxic organics and their metabolites in human biological fluids. Specifically, the method has played an important role in the highly sensitive and specific quantitative detection of persistent highly halogenated compounds in environmental matrices and biota during the past few decades. However, for the analysis of toxic metabolites with active hydrogen atoms, such as acids, alcohols, and phenolic compounds, from biological matrixes or organics without electronegative atoms or groups, a derivatization step is often needed prior to GC analysis. Such derivatization aims to change the properties of targets to improve their separation, increase their volatility, and enhance the sensitivity of instrumental detection. This review summarizes three derivatization strategies commonly used for GC methods, i.e., alkylation, silylation, and acylation, together with their application combined with GC-NCI-MS for the high sensitivity analysis of toxic organic metabolites in the human body. The advantages and disadvantages of each derivatization method and potential directions for future applications are discussed. Given the broad variety of applications as well as the compound-specific sensitivity for the ultra-trace analysis of target xenobiotics in human biological fluids, subsequent studies are required to develop convenient, faster derivatization procedures and reagents better suited for routine analysis.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Yu Z, Zheng K, Ren G, Zheng Y, Ma S, Peng P, et al. Identification of hydroxylated octa- and nona-bromodiphenyl ethers in human serum from electronic waste dismantling workers. Environ Sci Technol. 2010;44:3979–85.
Ma S, Ren G, Zeng X, Yu Z, Sheng G, Fu J. Polychlorinated biphenyls and their hydroxylated metabolites in the serum of e-waste dismantling workers from eastern China. Environ Geochem Health. 2018;40:1931–40.
Grova N, Hardy EM, Faÿs F, Duca RC, Appenzeller BMR. Hair analysis for the biomonitoring of polycyclic aromatic hydrocarbon exposure: comparison with urinary metabolites and DNA adducts in a rat model. Arch Toxicol. 2018;92:3061–75.
Hilton DC, Trinidad DA, Hubbard K, Li Z, Sjödin A. Measurement of urinary benzo[a]pyrene tetrols and their relationship to other polycyclic aromatic hydrocarbon metabolites and cotinine in humans. Chemosphere. 2017;189:365–72.
Wang Y, Liu S, Zhao H, Zhao G, Chen J, Zhai G, et al. Polybrominated diphenylethers (PBDEs) and their hydroxylated metabolites (OH-PBDEs) in female serum from Dalian, China. Int J Hyg Environ Health. 2016;219:816–22.
Hecht SS, Carmella SG, Villalta PW, Hochalter JB. Analysis of phenanthrene and benzo[a]pyrene tetraol enantiomers in human urine: relevance to the bay region diol epoxide hypothesis of benzo[a]pyrene carcinogenesis and to biomarker studies. Chem Res Toxicol. 2010;23:900–8.
Barbeau D, Lutier S, Choisnard L, Marques M, Persoons R, Maitre A. Urinary trans-anti-7,8,9,10-tetrahydroxy-7,8,9,10-tetrahydrobenzo(a)pyrene as the most relevant biomarker for assessing carcinogenic polycyclic aromatic hydrocarbons exposure. Environ Int. 2018;112:147–55.
Butryn DM, Gross MS, Chi LH, Schecter A, Olson JR, Aga DS. “One-shot” analysis of polybrominated diphenyl ethers and their hydroxylated and methoxylated analogs in human breast milk and serum using gas chromatography-tandem mass spectrometry. Anal Chim Acta. 2015;892:140–7.
Provencher G, Bérubé R, Dumas P, Bienvenu JF, Gaudreau É, Bélanger P, et al. Determination of bisphenol A, triclosan and their metabolites in human urine using isotope-dilution liquid chromatography–tandem mass spectrometry. J Chromatogr A. 2014;1348:97–104.
Tsikas D, Zoerner AA. Analysis of eicosanoids by LC-MS/MS and GC-MS/MS: a historical retrospect and a discussion. J Chromatogr B. 2014;964:79–88.
Dirtu AC, Roosens L, Geens T, Gheorghe A, Neels H, Covaci A. Simultaneous determination of bisphenol A, triclosan, and tetrabromobisphenol A in human serum using solid-phase extraction and gas chromatography-electron capture negative-ionization mass spectrometry. Anal Bioanal Chem. 2008;391:1175–81.
Lacorte S, Ikonomou MG. Occurrence and congener specific profiles of polybrominated diphenyl ethers and their hydroxylated and methoxylated derivatives in breast milk from Catalonia. Chemosphere. 2009;74:412–20.
Tsikas D. Pentafluorobenzyl bromide—a versatile derivatization agent in chromatography and mass spectrometry: I. Analysis of inorganic anions and organophosphates. J Chromatogr B. 2017;1043:187–201.
Gross JH. Chemical ionization. In: Mass spectrometry: a textbook. Springer International Publishing: Cham; 2017. p. 439–96.
Athanasiadou M, Cuadra SN, Marsh G, Bergman Å, Jakobsson K. Polybrominated diphenyl ethers (PBDEs) and bioaccumulative hydroxylated PBDE metabolites in young humans from Managua, Nicaragua. Environ Health Perspect. 2008;116:400–8.
Moldoveanu SC, David V. Derivatization methods in GC and GC/MS. In: Kusch P, editor. Gas chromatography - derivatization, sample preparation, application. IntechOpen; 2018. https://doi.org/10.5772/intechopen.81954.
Rohloff J. Analysis of phenolic and cyclic compounds in plants using derivatization techniques in combination with GC-MS-based metabolite profiling. Molecules. 2015;20:3431–62.
Koek MM, Jellema RH, van der Greef J, Tas AC, Hankemeier T. Quantitative metabolomics based on gas chromatography mass spectrometry: status and perspectives. Metabolomics. 2011;7:307–28.
Grova N, Hardy EM, Meyer P, Appenzeller BMR. Analysis of tetrahydroxylated benzo[a]pyrene isomers in hair as biomarkers of exposure to benzo[a]pyrene. Anal Bioanal Chem. 2016;408:1997–2008.
Langlois I, Berger U, Zencak Z, Oehme M. Mass spectral studies of perfluorooctane sulfonate derivatives separated by high-resolution gas chromatography. Rapid Commun Mass Spectrom. 2007;21:3547–53.
Zhai C, Peng S, Yang L, Wang Q. Evaluation of BDE-47 hydroxylation metabolic pathways based on a strong electron-withdrawing pentafluorobenzoyl derivatization gas chromatography/electron capture negative ionization quadrupole mass spectrometry. Environ Sci Technol. 2014;48:8117–26.
Bowden JA, Ford DA. An examination of pentafluorobenzoyl derivatization strategies for the analysis of fatty alcohols using gas chromatography/electron capture negative ion chemical ionization–mass spectrometry. J Chromatogr B. 2011;879:1375–83.
Winnberg U, Rydén A, Löfstrand K, Asplund L, Bignert A, Marsh G. Novel octabrominated phenolic diphenyl ether identified in blue mussels from the Swedish west coast. Environ Sci Technol. 2014;48:3319–26.
Zhao JL, Ying GG, Wang L, Yang JF, Yang XB, Yang LH, et al. Determination of phenolic endocrine disrupting chemicals and acidic pharmaceuticals in surface water of the Pearl Rivers in South China by gas chromatography–negative chemical ionization–mass spectrometry. Sci Total Environ. 2009;407:962–74.
Geens T, Neels H, Covaci A. Sensitive and selective method for the determination of bisphenol-A and triclosan in serum and urine as pentafluorobenzoate-derivatives using GC–ECNI/MS. J Chromatogr B. 2009;877:4042–6.
Oglobline AN, Elimelakh H, Tattam B, Geyer R, O’Donnell GE, Holder G. Negative ion chemical ionization GC/MS-MS analysis of dialkylphosphate metabolites of organophosphate pesticides in urine of non-occupationally exposed subjects. Analyst. 2001;126:1037–41.
Schindler BK, Förster K, Angerer J. Determination of human urinary organophosphate flame retardant metabolites by solid-phase extraction and gas chromatography–tandem mass spectrometry. J Chromatogr B. 2009;877:375–81.
Grova N, Salquèbre G, Hardy EM, Schroeder H, Appenzeller BMR. Tetrahydroxylated-benzo[a]pyrene isomer analysis after hydrolysis of DNA-adducts isolated from rat and human white blood cells. J Chromatogr A. 2014;1364:183–91.
Schummer C, Delhomme O, Appenzeller BMR, Wennig R, Millet M. Comparison of MTBSTFA and BSTFA in derivatization reactions of polar compounds prior to GC/MS analysis. Talanta. 2009;77:1473–82.
Xiao X, McCalley D. Quantitative analysis of estrogens in human urine using gas chromatography/negative chemical ionisation mass spectrometry. Rapid Commun Mass Spectrom. 2000;14:1991–2001.
Choi MH, Chung BC, Lee W, Lee UC, Kim Y. Determination of anabolic steroids by gas chromatography/negative-ion chemical ionization mass spectrometry and gas chromatography/negative-ion chemical ionization tandem mass spectrometry with heptafluorobutyric anhydride derivatization. Rapid Commun Mass Spectrom. 1999;13:376–80.
Fiamegos YC, Stalikas CD. Gas chromatographic determination of amino acids via one-step phase-transfer catalytic pentafluorobenzylation–preconcentration. J Chromatogr A. 2006;1110:66–72.
Malmvärn A, Zebühr Y, Kautsky L, Bergman Å, Asplund L. Hydroxylated and methoxylated polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins in red alga and cyanobacteria living in the Baltic Sea. Chemosphere. 2008;72:910–6.
Fujii Y, Nishimura E, Kato Y, Harada KH, Koizumi A, Haraguchi K. Dietary exposure to phenolic and methoxylated organohalogen contaminants in relation to their concentrations in breast milk and serum in Japan. Environ Int. 2014;63:19–25.
Gebbink WA, Sonne C, Dietz R, Kirkegaard M, Riget FF, Born EW, et al. Tissue-specific congener composition of organohalogen and metabolite contaminants in East Greenland polar bears (Ursus maritimus). EnvironPollut. 2008;152:621–9.
Henderson WM, Weber EJ, Duirk SE, Washington JW, Smith MA. Quantification of fluorotelomer-based chemicals in mammalian matrices by monitoring perfluoroalkyl chain fragments with GC/MS. J Chromatogr B. 2007;846:155–61.
Schoental R. Carcinogenic action of diazomethane and of nitroso-N-methyl urethane. Nature. 1960;188:420–1.
Mateo-Vivaracho L, Cacho J, Ferreira V. Quantitative determination of wine polyfunctional mercaptans at nanogram per liter level by gas chromatography–negative ion mass spectrometric analysis of their pentafluorobenzyl derivatives. J Chromatogr A. 2007;1146:242–50.
Tsikas D, Rothmann S, Schneider JY, Suchy M-T, Trettin A, Modun D, et al. Development, validation and biomedical applications of stable-isotope dilution GC–MS and GC–MS/MS techniques for circulating malondialdehyde (MDA) after pentafluorobenzyl bromide derivatization: MDA as a biomarker of oxidative stress and its relation to 15(S)-8-iso-prostaglandin F2α and nitric oxide (NO). J Chromatogr B. 2016;1019:95–111.
Brock JW, Yoshimura Y, Barr JR, Maggio VL, Graiser SR, Nakazawa H, et al. Measurement of bisphenol A levels in human urine. J Expo Sci Environ Epidemiol. 2001;11:323–8.
Pagliano E, Campanella B, D'Ulivo A, Mester Z. Derivatization chemistries for the determination of inorganic anions and structurally related compounds by gas chromatography - a review. Anal Chim Acta. 2018;1025:12–40.
Kage S, Kudo K, Ikeda N. Simultaneous determination of nitrate and nitrite in human plasma by gas chromatography-mass spectrometry. J Anal Toxicol. 2002;26:320–4.
Chen Z, Maartens F, Vega H, Kunene S, Gumede J, Krieger RI. 2,2-bis(4-chlorophenyl)acetic acid (DDA), a water-soluble urine biomarker of DDT metabolism in humans. Int J Toxicol. 2009;28:528–33.
Nakamura S, Takino M, Daishima S. Trace level determination of phenols as pentafluorobenzyl derivatives by gas chromatography–negative-ion chemical ionization mass spectrometry. Analyst. 2001;126:835–9.
Kuch HM, Ballschmiter K. Determination of endocrine-disrupting phenolic compounds and estrogens in surface and drinking water by HRGC−(NCI)−MS in the picogram per liter range. Environ Sci Technol. 2001;35:3201–6.
Allmyr M, McLachlan MS, Sandborgh-Englund G, Adolfsson-Erici M. Determination of triclosan as its pentafluorobenzoyl ester in human plasma and milk using electron capture negative ionization mass spectrometry. Anal Chem. 2006;78:6542–6.
Bravo R, Caltabiano LM, Weerasekera G, Whitehead RD, Fernandez C, Needham LL, et al. Measurement of dialkyl phosphate metabolites of organophosphorus pesticides in human urine using lyophilization with gas chromatography-tandem mass spectrometry and isotope dilution quantification. J Expo Sci Environ Epidemiol. 2004;14:249–59.
Myers SR, Ali Y. Determination of tobacco specific hemoglobin adducts in smoking mothers and new born babies by mass spectrometry. Biomark Insights. 2007;2:269–82.
Fitzgerald RL, Rexin DA, Herold DA. Benzodiazepine analysis by negative chemical ionization gas chromatography/mass spectrometry. J Anal Toxicol. 1993;17:342–7.
Campbell JA, Timchalk C, Kousba AA, Wu H, Valenzuela BR, Hoppe EW. Negative ion chemical ionization mass spectrometry for the analysis of 3,5,6-trichloro-2-pyridinol in saliva of rats exposed to chlorpyrifos. Anal Lett. 2005;38:939–49.
Boysen G, Hecht SS. Analysis of DNA and protein adducts of benzo[a]pyrene in human tissues using structure-specific methods. Mutat Res Rev Mutat Res. 2003;543:17–30.
Hecht SS, Chen M, Yagi H, Jerina DM, Carmella SG. r-1,t-2,3,c-4-Tetrahydroxy-1,2,3,4-tetrahydrophenanthrene in human urine: a potential biomarker for assessing polycyclic aromatic hydrocarbon metabolic activation. Cancer Epidemiol Biomark Prev. 2003;12:1501–8.
Zhong Y, Carmella SG, Hochalter JB, Balbo S, Hecht SS. Analysis of r-7,t-8,9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene in human urine: a biomarker for directly assessing carcinogenic polycyclic aromatic hydrocarbon exposure plus metabolic activation. Chem Res Toxicol. 2011;24:73–80.
Simpson CD, Wu MT, Christiani DC, Santella RM, Carmella SG, Hecht SS. Determination of r-7,t-8,9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene in human urine by gas chromatography/negative ion chemical ionization/mass spectrometry. Chem Res Toxicol. 2000;13:271–80.
Grova N, Antignac J-P, Hardy EM, Monteau F, Pouponneau K, Le Bizec B, et al. Identification of new tetrahydroxylated metabolites of polycyclic aromatic hydrocarbons in hair as biomarkers of exposure and signature of DNA adduct levels. Anal Chim Acta. 2017;995:65–76.
Simpson JT, Torok DS, Markey SP. Pentafluorobenzyl chloroformate derivatization for enhancement of detection of amino acids or alcohols by electron capture negative ion chemical ionization mass spectrometry. J Am Soc Mass Spectrom. 1995;6:525–8.
Wong J-M T, Malec PA, Mabrouk OS, Ro J, Dus M, Kennedy RT. Benzoyl chloride derivatization with liquid chromatography–mass spectrometry for targeted metabolomics of neurochemicals in biological samples. J Chromatogr A. 2016;1446:78–90.
Lerch O, Zinn P. Derivatisation and gas chromatography–chemical ionisation mass spectrometry of selected synthetic and natural endocrine disruptive chemicals. J Chromatogr A. 2003;991:77–97.
Silvério ACP, Machado SC, Boralli VB, Martins I. Dialkyl phosphates determination by gas chromatography: evaluation of a microwave-assisted derivatization. J Sep Sci. 2015;38:2664–9.
Zhao LJ, Ni Y, Su MM, Li HS, Dong FC, Chen WL, et al. High throughput and quantitative measurement of nicrobial metabolome by gas chromatography/mass spectrometry using automated alkyl chloroformate derivatization. Anal Chem. 2017;89:5565–77.
Söderholm SL, Damm M, Kappe CO. Microwave-assisted derivatization procedures for gas chromatography/mass spectrometry analysis. Mol Divers. 2010;14:869–88.
Xu X, Zhao X, Zhang Y, Li D, Su R, Yang Q, et al. Microwave-accelerated derivatization prior to GC-MS determination of sex hormones. J Sep Sci. 2011;34:1455–62.
Bowden JA, Colosi DM, Stutts WL, Mora-Montero DC, Garrett TJ, Yost RA. Enhanced analysis of steroids by gas chromatography/mass spectrometry using microwave-accelerated derivatization. Anal Chem. 2009;81:6725–34.
Kieliba T, Lerch O, Andresen-Streichert H, Rothschild MA, Beike J. Simultaneous quantification of THC-COOH, OH-THC, and further cannabinoids in human hair by gas chromatography–tandem mass spectrometry with electron ionization applying automated sample preparation. Drug Test Anal. 2019;11:267–78.
Abbiss H, Rawlinson C, Maker GL, Trengove R. Assessment of automated trimethylsilyl derivatization protocols for GC-MS-based untargeted metabolomic analysis of urine. Metabolomics. 2015;11:1908–21.
This study was supported by the National Natural Science Foundation of China (41991310, 41703092, and 41907299), Local Innovative and Research Teams Project of the Guangdong Pearl River Talents Program (2017BT01Z032).
Conflict of interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Yang, Y., Lin, M., Tang, J. et al. Derivatization gas chromatography negative chemical ionization mass spectrometry for the analysis of trace organic pollutants and their metabolites in human biological samples. Anal Bioanal Chem (2020). https://doi.org/10.1007/s00216-020-02762-x
- Negative chemical ionization