A rapid screening color test for glyphosate using dabsyl derivatization

  • Daisuke WatanabeEmail author
  • Yuko Kazui
  • Hikoto Ohta
Short Communication



Glyphosate is the most common herbicide in the world, and involved in many poisoning cases. However, the analysis of glyphosate has been difficult because of its high polarity and zwitterionic structure, and no simple screening tests for glyphosate have been known. The purpose of this study was to develop a rapid screening test for glyphosate.


Glyphosate was visualized through a color change induced by dabsyl derivatization, and the derivatized glyphosate was trapped on the surface of zirconia-coated silica particles by specific chemical interaction between a phosphonate group of glyphosate and the zirconia surface. The presence of glyphosate was indicated by the color change of zirconia surface, which was easily noticed by the naked eye.


The developed method was applied to biological samples (urine and serum), beverage samples (green tea and cola drink), and soil samples. No false positive was observed and the detection limit was 100 μg/mL for the biological samples and 800 μg/mL for the beverage samples.


A simple screening method for glyphosate based on a color test was established. The method is rapid and requires less than 30 μL of samples. The sensitivity was sufficient for most cases of moderate to severe poisoning.


Color test Dabsyl chloride Glyphosate Screening test Zirconia-coated cartridge 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Glyphosate Technical Fact Sheet, National Pesticide Information Center. Accessed 3 Dec 2018.
  2. 2.
    Stalikas CD, Konidari CN (2001) Analytical methods to determine phosphonic and amino acid group-containing pesticides. J Chromatogr A 907:1–19CrossRefGoogle Scholar
  3. 3.
    Kudzin ZH, Gralak DK, Andrijewski G, Drabowicz J, Łuczak J (2003) Simultaneous analysis of biologically active aminoalkanephosphonic acids. J Chromatogr A 998:183–199CrossRefGoogle Scholar
  4. 4.
    Moye HA, Deyrup CL (1984) A simple single-step derivatization method for the gas chromatographic analysis of the herbicide glyphosate and its metabolite. J Agric Food Chem 32:192–195CrossRefGoogle Scholar
  5. 5.
    Hori Y, Fujisawa M, Shimada K, Hirose Y (2003) Determination of the herbicide glyphosate and its metabolite in biological specimens by gas chromatography-mass spectrometry. A case of poisoning by Roundup® herbicide. J Anal Toxicol 27:162–166CrossRefGoogle Scholar
  6. 6.
    Tsunoda N (1993) Simultaneous determination of the herbicides glyphosate, glufosinate and bialaphos and their metabolites by capillary gas chromatography-ion-trap mass spectrometry. J Chromatogr A 637:167–173CrossRefGoogle Scholar
  7. 7.
    Motojyuku M, Saito T, Akieda K, Otsuka H, Yamamoto I, Inokuchi S (2008) Determination of glyphosate, glyphosate metabolites, and glufosinate in human serum by gas chromatography-mass spectrometry. J Chromatogr B 875:509–514CrossRefGoogle Scholar
  8. 8.
    Stalikas CD, Pilidis GA (2000) Development of a method for the simultaneous determination of phosphoric and amino acid group containing pesticides by gas chromatography with mass-selective detection. Optimization of the derivatization procedure using an experimental design approach. J Chromatogr A 872:215–225CrossRefGoogle Scholar
  9. 9.
    Tomita M, Okuyama T (1991) High-performance liquid chromatographic determination of glyphosate and (aminomethyl)phosphonic acid in human serum after conversion into p-toluenesulphonyl derivatives. J Chromatogr 566:239–243CrossRefGoogle Scholar
  10. 10.
    Nedelkoska TV, Low GK-C (2004) High-performance liquid chromatographic determination of glyphosate in water and plant material after pre-column derivatisation with 9-fluorenylmethyl chloroformate. Anal Chim Acta 511:145–153CrossRefGoogle Scholar
  11. 11.
    Patsias J, Papadopoulou A, Papadopoulou-Mourkidou E (2001) Automated trace level determination of glyphosate and aminomethyl phosphonic acid in water by on-line anion-exchange solid-phase extraction followed by cation-exchange liquid chromatography and post-column derivatization. J Chromatogr A 932:83–90CrossRefGoogle Scholar
  12. 12.
    Bernal J, Bernal JL, Martin MT, Nozal MJ, Anadón A, Martínez-Larrañaga MR, Martínez MA (2010) Development and validation of a liquid chromatography-fluorescence-mass spectrometry method to measure glyphosate and aminomethylphosphonic acid in rat plasma. J Chromatogr B 878:3290–3296CrossRefGoogle Scholar
  13. 13.
    Yoshioka N, Asano M, Kuse A, Mitsuhashi T, Nagasaki Y, Ueno Y (2011) Rapid determination of glyphosate, glufosinate, bialaphos, and their major metabolites in serum by liquid chromatography-tandem mass spectrometry using hydrophilic interaction chromatography. J Chromatogr A 1218:3675–3680CrossRefGoogle Scholar
  14. 14.
    Guo H, Wang H, Zheng J, Liu W, Zhong J, Zhao Q (2018) Sensitive and rapid determination of glyphosate, glufosinate, bialaphos and metabolites by UPLC-MS/MS using a modified Quick Polar Pesticides Extraction method. Forensic Sci Int 283:111–117CrossRefGoogle Scholar
  15. 15.
    Watanabe D, Ohta H, Yamamuro T, Ohtsuru O, Miyazaki S, Ohira M (2014) Development of rapid cleanup method for phosphorous containing amino acid type herbicides by using monolithic silica spin column chemically modified by titanium dioxide, MonoSpin TiO. Rep Natl Res Inst Police Sci 63:1–7 (in Japanese) Google Scholar
  16. 16.
    Watanabe D, Ohta H, Yamamuro T (2014) Solid-phase extraction of phosphorous-containing amino acid herbicides from biological specimens with a zirconia-coated silica cartridge. J Chromatogr B 969:69–76CrossRefGoogle Scholar
  17. 17.
    Nawrocki J, Rigney MP, McCormick A, Carr PW (1993) Chemistry of zirconia and its use in chromatography. J Chromatogr A 657:229–282CrossRefGoogle Scholar
  18. 18.
    Romero R, Bagur MG, Sanchez-Vinas M, Gazquez D (2000) Optimization of experimental variables in the dabsyl chloride derivatization of biogenic amines for their determination by RP-HPLC. Chromatographia 51:404–410CrossRefGoogle Scholar
  19. 19.
    Chen Y-H, Shih L-L, Liou S-E, Chen C-C (2003) Analysis of dabsyl-Cl derivated amino acids by high performance liquid chromatography and tandem mass spectrometry. Food Sci Technol Res 9:276–282CrossRefGoogle Scholar
  20. 20.
    Zouaoui K, Dulaurent S, Gaulier JM, Moesch C, Lachâtre G (2013) Determination of glyphosate and AMPA in blood and urine from humans: About 13 cases of acute intoxication. Forensic Sci Int 226:e20–25CrossRefGoogle Scholar
  21. 21.
    Roberts DM, Buckley NA, Mohamed FM, Eddleston M, Goldstein DA, Mehrsheikh A, Bleeke MS, Dawson AH (2010) A prospective observational study of the clinical toxicology of glyphosate-containing herbicides in adults with acute self-poisoning. Clin. Toxicol (Phila) 48(2):129–136CrossRefGoogle Scholar
  22. 22.
    Suzuki O, Yashiki M (2002) Hand book of practical analysis of drugs and poisons in human specimens—chromatographic methods. Jiho, Tokyo, p 480 (in Japanese) Google Scholar

Copyright information

© Japanese Association of Forensic Toxicology 2018

Authors and Affiliations

  1. 1.National Research Institute of Police ScienceKashiwaJapan

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