Paper-based electrochemical sensor for on-site detection of the sulphur mustard

  • Noemi Colozza
  • Kai Kehe
  • Tanja Popp
  • Dirk Steinritz
  • Danila Moscone
  • Fabiana ArduiniEmail author
Innovations in environmental sciences related to chemical, biological, radiological and nuclear risks


Herein, we report a novel paper-based electrochemical sensor for on-site detection of sulphur mustards. This sensor was conceived combining office paper-based electrochemical sensor with choline oxidase enzyme to deliver a sustainable sensing tool. The mustard agent detection relies on the evaluation of inhibition degree of choline oxidase, which is reversibly inhibited by sulphur mustards, by measuring the enzymatic by-product H2O2 in chronoamperometric mode. A nanocomposite constituted of Prussian Blue nanoparticles and Carbon Black was used as working electrode modifier to improve the electroanalytical performances. This bioassay was successfully applied for the measurement of a sulphur mustard, Yprite, obtaining a detection limit in the millimolar range (LOD = 0.9 mM). The developed sensor, combined with a portable and easy-to-use instrumentation, can be applied for a fast and cost-effective detection of sulphur mustards.


Screen-printed electrode Prussian blue nanoparticles, carbon black Chemical warfare agents Choline oxidase, inhibition 


Funding information

F.A. acknowledges the Italian Ministry of Defence, Aptamer BW project for the financial support.


  1. Abe K, Suzuki K, Citterio D (2008) Inkjet-printed microfluidic multianalyte chemical sensing paper. Anal Chem 80:6928–6934CrossRefGoogle Scholar
  2. Arduini F, Palleschi G (2012) Disposable electrochemical biosensor based on cholinesterase inhibition with improved shelf-life and working stability for nerve agent detection. In: NATO Science for Peace and Security Series A: Chemistry and Biology Portable Chemical Sensors Weapons Against Bioterrorism, Springer, pp. 261–278Google Scholar
  3. Arduini F, Scognamiglio V, Covaia C, Amine A, Moscone D, Palleschi G (2015) A choline oxidase Amperometric bioassay for the detection of mustard agents based on screen-printed electrodes modified with Prussian blue nanoparticles. Sensors 15:4353–4367CrossRefGoogle Scholar
  4. Barron ESG, Bartlett GR, Miller ZB, Meyer J, Seegmiller JE (1948) The effect of nitrogen mustards on enzymes and tissue metabolism. J Exp Med 6:503–519CrossRefGoogle Scholar
  5. Brookes P, Lawley PD (1961) The reaction of mono-and di-functional alkylating agents with nucleic acids. Biochem J 80:496–503CrossRefGoogle Scholar
  6. Cinti S, Arduini F, Vellucci G, Cacciotti I, Nanni F, Moscone D (2014) Carbon black assisted tailoring of Prussian blue nanoparticles to tune sensitivity and detection limit towards H2O2 by using screen-printed electrode. Electrochem Commun 47:63–66CrossRefGoogle Scholar
  7. Cinti S, Talarico D, Palleschi G, Moscone D, Arduini F (2016) Novel reagentless paper-based screen-printed electrochemical sensor to detect phosphate. Anal Chim Acta 919:78–84CrossRefGoogle Scholar
  8. Cinti S, Minotti C, Moscone D, Palleschi G, Arduini F (2017a) Fully integrated ready-to-use paper-based electrochemical biosensor to detect nerve agents. Biosens Bioelectron 93:46–51CrossRefGoogle Scholar
  9. Cinti S, Basso M, Moscone D, Arduini F (2017b) A paper-based nanomodified electrochemical biosensor for ethanol detection in beers. Anal Chim Acta 960:123–130CrossRefGoogle Scholar
  10. Dungchai W, Chailapakul O, Henry CS (2009) Electrochemical detection for paper-based microfluidics. Anal Chem 81:5821–5826CrossRefGoogle Scholar
  11. Fu D, Calvo JA, Samson LD (2012) Balancing repair and tolerance of DNA damage caused by alkylating agents. Nat Rev Cancer 12:104–120CrossRefGoogle Scholar
  12. Ghanei M, Amiri S, Akbari H, Kosari F, Khalili AR, Alaeddini F, Aslani J, Giardina C, Haines DD (2007) Correlation of sulfur mustard exposure and tobacco use with expression (immunoreactivity) of p53 protein in bronchial epithelium of Iranian “mustard lung” patients. Mil Med 172:70–74CrossRefGoogle Scholar
  13. Haines DD, Fox SC (2014) Acute and long-term impact of chemical weapons: lessons from the Iran-Iraq war. Forensic Sci Rev 26:97–114Google Scholar
  14. Hill HH, Martin SJ (2002) Conventional analytical methods for chemical warfare agents. Pure Appl Chem 74:2281–2291CrossRefGoogle Scholar
  15. International Agency for Research on Cancer (IARC): IARC monograph on the evaluation of carcinogenic risks to humans; WHO Press: Geneva, Switzerland; 2004; http:// (accessed January 31, 2018)
  16. Karyakin AA, Karyakina EE, Gorton L (1996) Prussian-blue-based amperometric biosensors in flow-injection analysis. Talanta 43:1597–1606CrossRefGoogle Scholar
  17. Kehe K, Szinicz L (2005) Medical aspects of sulphur mustard poisoning. Toxicology 214:198–209CrossRefGoogle Scholar
  18. Kehe K, Balszuweit F, Emmler J, Kreppel H, Jochum M, Thiermann H (2008) Sulfur mustard research—strategies for the development of improved medical therapy. Eplasty 8Google Scholar
  19. Kehe K, Balszuweit F, Steinritz D, Thiermann H (2009) Molecular toxicology of sulfur mustard-induced cutaneous inflammation and blistering. Toxicology 263:12–19CrossRefGoogle Scholar
  20. Khateri S, Ghanei M, Soroush MR, Haines DD (2003a) Effects of mustard gas exposure in pediatric patients: long-term health status of mustard-exposed children 14 years after chemical bombardment of Sardasht. J Burns Surg Wound Care 2:11Google Scholar
  21. Khateri S, Ghanei S, Keshavarz M, Soroush D, Haines DD (2003b) Incidence of lung, eye and skin lesions as late complications in 34,000 Iranians with wartime exposure to mustard agent. J Occup Environ Med 45:1136–1143CrossRefGoogle Scholar
  22. Komkova MA, Karyakina EE, Karyakin AA (2017) Noiseless performance of Prussian blue based (bio) sensors through power generation. Anal Chem 89:6290–6294CrossRefGoogle Scholar
  23. Kumar V, Anslyn EV (2013) A selective turn-on fluorescent sensor for sulfur mustard simulants. J Am Chem Soc 135:6338–6344CrossRefGoogle Scholar
  24. López-Marzo AM, Merkoçi A (2016) Paper-based sensors and assays: a success of the engineering design and the convergence of knowledge areas. Lab Chip 16:3150–3176CrossRefGoogle Scholar
  25. Martinez AW, Phillips ST, Butte MJ, Whitesides GM (2007) Patterned paper as a platform for inexpensive, low volume, portable bioassays. Angew Chem Int Ed 46:1318–1320CrossRefGoogle Scholar
  26. McGill RA, Nguyen VK, Chung R, Shaffer RE, DiLella D, Stepnowski JL, Dominguez D (2000) The “NRL-SAWRHINO”: a nose for toxic gases. Sensors Actuators B 65:10–13CrossRefGoogle Scholar
  27. Menger FM, Elrington AR (1991) Organic reactivity in microemulsion systems. JACS 113:9621–9624CrossRefGoogle Scholar
  28. Meredith NA, Quinn C, Cate DM, Reilly TH, Volckens J, Henry CS (2016) Paper based analytical devices for environmental analysis. Analyst 141:1874–1887CrossRefGoogle Scholar
  29. Mettakoonpitak J, Boehle K, Nantaphol S, Teengam P, Adkins JA, Srisa-Art M, Henry CS (2016) Electrochemistry on paper-based analytical devices: a review. Electroanalysis 28:1420–1436CrossRefGoogle Scholar
  30. Moscone D, D'ottavi D, Compagnone D, Palleschi G, Amine A (2001) Construction and analytical characterization of Prussian blue-based carbon paste electrodes and their assembly as oxidase enzyme sensors. Anal Chem 73:2529–2535CrossRefGoogle Scholar
  31. Notingher I (2007) Raman spectroscopy cell-based biosensors. Sensors 7:1343–1358CrossRefGoogle Scholar
  32. Pitschmann V (2014) Overall view of chemical and biochemical weapons. Toxins 6:1761–1784CrossRefGoogle Scholar
  33. Reddy MK, Mills G, Nixon C, Wyatt SA, Croley TR (2011) High-throughput sample preparation and simultaneous column regeneration liquid chromatography–tandem mass spectrometry method for determination of nitrogen mustard metabolites in human urine. J Chromatogr B 879:2383–2388CrossRefGoogle Scholar
  34. Ricci F, Palleschi G (2005) Sensor and biosensor preparation, optimisation and applications of Prussian blue modified electrodes. Biosens Bioelectron 21:389–407CrossRefGoogle Scholar
  35. Ricci F, Amine A, Palleschi G, Moscone D (2003) Prussian blue based screen printed biosensors with improved characteristics of long-term lifetime and pH stability. Biosens Bioelectron 18:165–174CrossRefGoogle Scholar
  36. Roshan R, Rahnama P, Ghazanfari Z, Montazeri A, Soroush MR, Naghizadeh MM, Melyani M, Tavoli A, Ghazanfari T (2013) Long-term effects of sulfur mustard on civilians’ mental health 20 years after exposure (the Sardasht-Iran cohort study). Health Qual Life Outcomes 11:69–76CrossRefGoogle Scholar
  37. Rowell M, Kehe K, Balszuweit F, Thiermann H (2009) The chronic effects of sulfur mustard exposure. Toxicology 263:9–11CrossRefGoogle Scholar
  38. Salamati P, Razavi SM, Shokraneh F, Mohazzab S, Laal M, Hadjati G, Khaji A, Rahimi Movaghar V (2013) Mortality and injuries among Iranians in Iraq-Iran war: a systematic review. Arc Iran Med 16:542–550Google Scholar
  39. Salazar P, Martín M, O’Neill RD, Roche R, González-Mora JL (2012) Improvement and characterization of surfactant-modified Prussian blue screen-printed carbon electrodes for selective H2O2 detection at low applied potentials. J Electroanal Chem 674:48–56CrossRefGoogle Scholar
  40. Sanders CA, Rodriguez M, Greenbaum E (2001) Stand-off tissue-based biosensors for the detection of chemical warfare agents using photosynthetic fluorescence induction. Biosens Bioelectron 16:439–446CrossRefGoogle Scholar
  41. Shohrati M, Davoudi M, Ghanei M, Peyman M, Peyman A (2007) Cutaneous and ocular late complications of sulfur mustard in Iranian veterans. Cutan Ocul Toxicol 26:73–81CrossRefGoogle Scholar
  42. Singh VV, Nigam AK, Boopathi M, Pandey P, Singh B, Vijayaraghavan R (2012) In situ electrochemical sensing of 2-chloroethyl ethyl sulfide (CEES) a CWA simulant using CuPc/RTIL composite gold electrode. Sensors Actuators B 161:1000–1009CrossRefGoogle Scholar
  43. Singh VV, Nigam AK, Yadav SS, Tripathi BK, Srivastava A, Boopathi M, Singh B (2013) Graphene oxide as carboelectrocatalyst for in situ electrochemical oxidation and sensing of chemical warfare agent simulant. Sensors Actuators B 188:1218–1224CrossRefGoogle Scholar
  44. Steinritz D, Emmler J, Hintz M, Worek F, Kreppel H, Szinicz L, Kehe K (2007) Apoptosis in sulfur mustard treated A549 cell cultures. Life Sci 80:2199–2201CrossRefGoogle Scholar
  45. Takeshima Y, Inai K, Bennett WP, Metcalf RA, Welsh JA, Yonehara S, Hayashi Y, Fujihara M, Yamakido M, Akiyama M, Tokuoka S, Land CE, Harris CC (1994) p53 mutations in lung cancers from Japanese mustard gas workers. Carcinogenesis 15:2075–2079CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Noemi Colozza
    • 1
  • Kai Kehe
    • 2
  • Tanja Popp
    • 3
    • 4
  • Dirk Steinritz
    • 3
    • 4
  • Danila Moscone
    • 1
  • Fabiana Arduini
    • 1
    Email author
  1. 1.Department of Chemical Science and TechnologiesUniversity of Rome Tor VergataRomeItaly
  2. 2.Bundeswehr Medical AcademyMedical CBRN DefenseMunichGermany
  3. 3.Bundeswehr Institute of Pharmacology and ToxicologyMunichGermany
  4. 4.Walther-Straub-Institute of Pharmacology and ToxicologyLudwig-Maximilians-UniversitätMunichGermany

Personalised recommendations