Environmental Science and Pollution Research

, Volume 25, Issue 11, pp 11074–11083 | Cite as

The occurrence of selected xenobiotics in the Danube river via LC-MS/MS

  • Nataša Milić
  • Maja Milanović
  • Jelena Radonić
  • Maja Turk Sekulić
  • Anamarija Mandić
  • Dejan Orčić
  • Aleksandra Mišan
  • Ivan Milovanović
  • Nevena Grujić Letić
  • Mirjana Vojinović Miloradov
Research Article


Having in mind that there is a general lack of monitoring plans and precaution measures in the developing countries and that the Danube is the second longest river in Europe, the estimation of the relevant concentration levels of unregulated xenobiotics is a topic of interest both on local and international level. The selected pharmaceuticals, caffeine, and benzotriazole presented in the collected water samples from seven representative locations around the territory of Novi Sad, Serbia, during 1-year period, were analyzed with the use of solid-phase extraction followed by the liquid chromatography coupled with triple quad tandem mass spectrometry. The most frequently detected compounds were caffeine and carbamazepine in the concentrations up to 621 and 22.2 ng/L, respectively, while the maximum concentration of the analyzed pharmaceuticals was obtained for ibuprofen (60.1 ng/L). The presence of benzotriazole along the analyzed section of the river was confirmed in the concentration levels up to 26.7 ng/L. Although sulfamethoxazole and desmethyldiazepam were detected at trace levels (0.22 and 3.41 ng/L, respectively); the presence of these pharmaceuticals in complex mixtures should not be neglected. Due to the frequent detection caffeine, carbamazepine, ibuprofen, and benzotriazole could be proper candidate for hydrophilic anthropogenic markers for quantification of wastewater contamination in surface water in the analyzed Danube section.


The Danube Pharmaceuticals Emerging substances Environmental analysis Liquid chromatography-mass spectrometry Water quality 



The work was financially supported by the Ministry of Education, Science and Technological Development, Republic of Serbia (III46009) and NATO Science for Peace Project (ESP.EAP.SFP 984087).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Alotaibi MD, McKinley AJ, Patterson BM, Reeder AY (2015) Benzotriazoles in the aquatic environment: a review of their occurrence, toxicity, degradation and analysis. Water Air Soil Pollut 226(7):1–20. CrossRefGoogle Scholar
  2. Andersson T, Miners JO, Veronese ME, Birkett DJ (1994) Diazepam metabolism by human liver microsomes is mediated by both S-mephenytoin hydroxylase and CYP3A isoforms. Br J Clin Pharmacol 38(13):1–137Google Scholar
  3. Asimakopoulos AG, Wang L, Thomaidis NS, Kannan K (2013) Benzotriazoles and benzothiazoles in human urine from several countries: a perspective on occurrence, biotransformation, and human exposure. Environ Int 59:274–281. CrossRefGoogle Scholar
  4. Baldessarini RJ (1996) Drugs and the treatment of psychiatric disorders: depression and mania. In: Hardman JG et al (eds) Goodman & Gilman’s the Pharmacological Basis of Therapeutics. McGraw-Hill, New YorkGoogle Scholar
  5. Banzhaf S, Krein A, Scheytt T (2013) Using selected pharmaceutical compounds as indicators for surface water and groundwater interaction in the hyporheic zone of a low permeability riverbank. Hydrol Process 27:2892–2902. Google Scholar
  6. Barton BA, Morgan JD, Vijayan MM (2002) Physiological and condition-related indicators of environmental stress in fish. In: Adams (ed) Biological Indicatorof Aquatic Ecosystem Stress. American Fisheries Society, VancouverGoogle Scholar
  7. Buser HR, Poiger T, Muller MD (1999) Occurrence and environmental behavior of the chiral pharmaceutical drug ibuprofen in surface waters and in wastewater. Environ Sci Technol 33:2529–2535. CrossRefGoogle Scholar
  8. Caliman FA, Gavrilescu M (2009) Pharmaceuticals, personal care products and endocrine disrupting agents in the environment—a review. CLEAN Soil Air Water 37(4-5):277–303. CrossRefGoogle Scholar
  9. Chițescu CL, Nicolau AI (2014) Preliminary survey of pharmaceutical residues in some important Romanian rivers. Toxicol Environ Chem 96(9):1333–1345. CrossRefGoogle Scholar
  10. Dulio V, Slobodnik J (2009) NORMAN—network of reference laboratories, research centres and related organisations for monitoring of emerging substances. Environ Sci Pollut Res 16(S1):132–135. CrossRefGoogle Scholar
  11. European Commission (2002) Commission Decision 2002/657/EC of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Comm L, pp 221Google Scholar
  12. European Commission (2015) Commission implementing decision (EU) 2015/495 of 20 March 2015 establishing a watch list of substances for Union-wide monitoring in the field of water policy pursuant to Directive 2008/105/EC of the European Parliament and of the Council. Off J Eur Union L78, pp 40–42Google Scholar
  13. Ferrando-Climent L, Collado N, Buttiglieri G, Gros M, Rodriguez-Roda I, Rodriguez-Mozaz S, Barceló D (2012) Comprehensive study of ibuprofen and its metabolites in activated sludge batch experiments and aquatic environment. Sci Total Environ 438:404–413. CrossRefGoogle Scholar
  14. Giacomini ACVV, Abreu MS, Giacomini LV, Siebel AM, Zimerman FF, Rambo CL, Mocelin R, Bonan CD, Piato AL, Barcellos LJG (2016) Fluoxetine and diazepam acutely modulate stress induced-behavior. Behav Brain Res 296:301–310. CrossRefGoogle Scholar
  15. Giger W, Schaffner C, Kohler HPE (2006) Benzotriazole and tolyltriazole as aquatic contaminants. 1. Input and occurrence in rivers and lakes. Environ Sci Technol 40:7186–7192. CrossRefGoogle Scholar
  16. Gonçalves ES, Rodrigues SV, da Silva-Filho EV (2016) The use of caffeine as a chemical marker of domestic wastewater contamination in surface waters: seasonal and spatial variations in Teresópolis, Brazil. Rev Ambient Água 12:192–202. CrossRefGoogle Scholar
  17. Gothwal R, Shashidhar T (2015) Antibiotic pollution in the environment: a review. CLEAN Soil Air Water 43:479–489CrossRefGoogle Scholar
  18. Gros M, Petrović M, Barceló D (2006) Development of a multi-residue analytical methodology based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) for screening and trace level determination of pharmaceuticals in surface and wastewaters. Talanta 70:678–690CrossRefGoogle Scholar
  19. Grujić Letić N, Milanović M, Milić N, Vojinović Miloradov M, Radonić J, Mihajlović I, Turk Sekulić M (2015) Determination of emerging substances in the Danube and potential risk evaluation. CLEAN Soil Air Water 43(5):731–738. CrossRefGoogle Scholar
  20. Grujić S, Vasiljević T, Laušević M (2009) Determination of multiple pharmaceutical classes in surface and ground waters by liquid chromatography-ion trap-tandem mass spectrometry. J Chromatogr A 1216(25):4989–5000. CrossRefGoogle Scholar
  21. Harris CA, Routledge EJ, Schaffner C, Brian JV, Giger W, Sumpter JP (2007) Benzotriazole is antiestrogenic in vitro but not in vivo. Environ Toxicol Chem 26(11):2367–2372. CrossRefGoogle Scholar
  22. Health Council of The Netherlands: Dutch Expert Committee on Occupational Standards (DECOS) (2000) 1,2,3-Benzotriazole. Publication No. 2000/14OSHGoogle Scholar
  23. Jjemba PK (2006) Excretion and ecotoxicity of pharmaceutical and personal care products in the environment. Ecotoxicol Environ Saf 63(1):113–130. CrossRefGoogle Scholar
  24. Kahle M, Buerge IJ, Müller MD, Poiger T (2009) Hydrophilic anthropogenic markers for quantification of wastewater contamination in ground- and surface waters. Environ Toxicol Chem 28(12):2528–2536. CrossRefGoogle Scholar
  25. Kasprzyk-Hordern B, Dinsdale RM, Guwy AJ (2007) Multi-residue method for the determination of basic/neutral pharmaceuticals and illicit drugs in surface water by solid-phase extraction and ultra performance liquid chromatography-positive electrospray ionisation tandem mass spectrometry. J Chromatogr A 1161(1-2):132–145. CrossRefGoogle Scholar
  26. Kolpin D, Furlong E, Meyer M, Thurman E, Zaugg S, Barber L, Buxton H (2002) Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000: a national reconnaissance. Environ Sci Technol 36:1202–1211. CrossRefGoogle Scholar
  27. Kosjek T, Perko S, Zupanc M, Zanoški Hren M, Landeka Dragičević T, Žigon D, Kompare B, Heath E (2012) Environmental occurrence, fate and transformation of benzodiazepines in water treatment. Water Res 46(2):355–368. CrossRefGoogle Scholar
  28. Kovačević S, Radišić M, Laušević M, Dimkić M (2017) Occurrence and behavior of selected pharmaceuticals during riverbank filtration in The Republic of Serbia. Environ Sci Pollut Res 24(2):2075–2088. CrossRefGoogle Scholar
  29. Liška I, Wagner F, Slobodnik J (eds) (2008) Joint Danube Survey 2, Final Scientific Report, ICPDR- International Commission for the Protection of the Danube River, Vienna. Accessed 20 Jan 2018
  30. Liška I, Wagner F, Sengl M, Deutsch K, Slobodník S (eds) (2015) Joint Danube Survey 3, A Comprehensive Analysis of Danube Water Quality, ICPDR- International Commission for the Protection of the Danube River, Vienna. Accessed 20 Jan 2018
  31. Löffler D, Römbke J, Meller M, Ternes TA (2005) Environmental fate of pharmaceuticals in water/sediment systems. Environ Sci Technol 39(14):5209–5218. CrossRefGoogle Scholar
  32. Loos R, Wollgast J, Huber T, Hanke G (2007) Polar herbicides, pharmaceutical products, perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and nonylphenol and its carboxylates and ethoxylates in surface and tap waters around Lake Maggiore in Northern Italy. Anal Bioanal Chem 387(4):1469–1478. CrossRefGoogle Scholar
  33. Loos R, Locoro G, Contini S (2010) Occurrence of polar organic contaminants in the dissolved water phase of the Danube River and its major tributaries using SPE-LC-MS2 analysis. Water Res 44:2325–2335. CrossRefGoogle Scholar
  34. Miao XS, Metcalfe CD (2003) Determination of carbamazepine and its metabolites in aqueous samples using liquid chromatography–electrospray tandem mass spectrometry. Anal Chem 75(15):3731–3738. CrossRefGoogle Scholar
  35. Milanović M, Sudji J, Grujić Letić N, Radonić J, Turk Sekulić M, Vojinović Miloradov M, Milić N (2016) Seasonal variations of bisphenol A in the Danube by the Novi Sad municipality, Serbia. J Serb Chem Soc 80(3):333–345CrossRefGoogle Scholar
  36. Milić N, Milanović M, Grujić Letić N, Turk Sekulić M, Radonić J, Mihajlović I, Vojinović Miloradov M (2013) Occurrence of antibiotics as emerging contaminant substances in aquatic environment. Int J Environ Health Res 23:296–310. CrossRefGoogle Scholar
  37. Milić N, Spanik I, Radonić J, Turk Sekulić M, Grujić N, Vyviurska O, Milanović M, Sremački M, Vojinović Miloradov M (2014) Screening analyses of wastewater and Danube surface water in Novi Sad locality, Serbia. Fresenius Environ Bull 23:372–377Google Scholar
  38. Mompelat S, Le Bot B, Thomas O (2009) Occurrence and fate of pharmaceutical products and by-products, from resource to drinking water. Environ Int 35:803–814. CrossRefGoogle Scholar
  39. Neumeyer JL, Booth RG (1995) Neuroleptics and anxiolytic agents. In: Foye WO et al (eds) Principles of Medicinal Chemistry. Williams & Wilkins, PhiladelphiaGoogle Scholar
  40. Nödler K, Licha T, Barbieri M, Pérez S (2012) Evidence for the microbially mediated abiotic formation of reversible and non-reversible sulfamethoxazole transformation products during denitrification. Water Res 46(7):2131–2139. CrossRefGoogle Scholar
  41. Norton TR, Lazev AB, Sullivan MJ (2011) The “buzz” on caffeine: patterns of caffeine use in a convenience sample of college students. J Caffeine Res 1(1):35–40. CrossRefGoogle Scholar
  42. Petrović M, Škrbić B, Živančev J, Ferrando-Climent L, Barcelo D (2014) Determination of 81 pharmaceutical drugs by high performance liquid chromatography coupled to mass spectrometry with hybrid triple quadrupole-linear ion trap in different types of water in Serbia. Sci Total Environ 468-469:415–428. CrossRefGoogle Scholar
  43. Radović T, Grujić S, Petković A, Dimkić M, Laušević M (2015) Determination of pharmaceuticals and pesticides in river sediments and corresponding surface and ground water in the Danube River and tributaries in Serbia. Environ Monit Assess 187(1):4092 (pp1–17). CrossRefGoogle Scholar
  44. Reemtsma T, Weiss S, Mueller J, Petrovic M, González S, Barcelo D, Ventura F, Knepper TP (2006) Polar pollutants entry into the water cycle by municipal wastewater: a European perspective. Environ Sci Technol 40(17):5451–5458. CrossRefGoogle Scholar
  45. Reemtsma T, Miehe U, Duennbier U, Jekel M (2010) Polar pollutants in municipal wastewater and the water cycle: occurrence and removal of benzotriazoles. Water Res 44(2):596–604. CrossRefGoogle Scholar
  46. Smith-Kielland A, Skuterud B, Olsen KM, Morland J (2001) Urinary excretion of diazepam metabolites in healthy volunteers and drug users. Scand J Clin Lab Invest 61(3):237–246. CrossRefGoogle Scholar
  47. Spence PL (2015) Using Caffeine as a Water Quality Indicator in the Ambient Monitoring Program for Third Fork Creek Watershed, Durham, North Carolina. Environ Health Insights 9(s2):29–34. Google Scholar
  48. Tangtian H, Bo L, Wenhua L, Shin PKS, Wu R (2012) Estrogenic potential of benzotriazole on marine medaka (Oryzias melastigma). Ecotoxicol Environ Saf 80:327–332. CrossRefGoogle Scholar
  49. Terzić S, Senta I, Ahel M, Gros M, Petrovic M, Barcelo D, Müller J, Knepper T, Marti I, Ventura F, Jovančić P, Jabučar D (2008) Occurrence and fate of emerging wastewater contaminants in Western Balkan Region. Sci Total Environ 399(1-3):66–77. CrossRefGoogle Scholar
  50. U.S. Environmental Protection Agency (EPA) (2016) Method detection limit (MDL) procedure found in Title 40 Code of Federal Regulations Part 136 (40 CFR 136, Appendix B, Revision 2)Google Scholar
  51. van Leerdam JA, Hogenboom AC, van der Kooi MME, de Voogt P (2009) Determination of polar 1H-benzotriazoles and benzothiazoles in water by solid-phase extraction and liquid chromatography LTQ FT Orbitrap mass spectrometry. Int J Mass Spectrom 282(3):99–107. CrossRefGoogle Scholar
  52. Vojinović Miloradov MB, Turk Sekulić MM, Radonić JR, Milić NB, Grujić Letić NN, Mihajlović IJ, Milanović MLJ (2014a) Industrial emerging chemicals in the environment. Chem Ind 68(1):51–62. (in Serbian). CrossRefGoogle Scholar
  53. Vojinović Miloradov M, Mihajlović I, Vyviurska O, Cacho F, Radonić J, Milić N, Spanik I (2014b) Impact of wastewater discharges to Danube surface water pollution by emerging and priority pollutants in the vicinity of Novi Sad, Serbia. Fresenius Environ Bull 23(5):2137–2145. Google Scholar
  54. Vulliet E, Cren-Olivé C (2011) Screening of pharmaceuticals and hormones at the regional scale, in surface and ground waters intended to human consumption. Environ Pollut 159(10):2929–2934. CrossRefGoogle Scholar
  55. Wang L, Zhang J, Sun H, Zhou Q (2016) Widespread occurrence of benzotriazoles and benzothiazoles in tap water: influencing factors and contribution to human exposure. Environ Sci Technol 50(5):2709–2017. CrossRefGoogle Scholar
  56. Wendelaar Bonga SE (1997) The stress response in fish. Physiol Rev 77(3):591–625. CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Nataša Milić
    • 1
  • Maja Milanović
    • 1
  • Jelena Radonić
    • 2
  • Maja Turk Sekulić
    • 2
  • Anamarija Mandić
    • 3
  • Dejan Orčić
    • 4
  • Aleksandra Mišan
    • 3
  • Ivan Milovanović
    • 3
  • Nevena Grujić Letić
    • 1
  • Mirjana Vojinović Miloradov
    • 2
  1. 1.Faculty of MedicineUniversity of Novi SadNovi SadSerbia
  2. 2.Faculty of Technical SciencesUniversity of Novi SadNovi SadSerbia
  3. 3.Institute of Food TechnologyUniversity of Novi SadNovi SadSerbia
  4. 4.Faculty of SciencesUniversity of Novi SadNovi SadSerbia

Personalised recommendations