Protocols for Measurement of Naphthenic Acids in Aqueous Samples

  • Lisa Brown
  • Ania UlrichEmail author
Part of the Springer Protocols Handbooks book series (SPH)


Naphthenic acids (NAs), a group of polar organic carboxylic acids that naturally occur in crude oil, have been of concern since the turn of the twentieth century due to corrosion of petroleum refineries. In the last two decades, naphthenic acids have been a focus of concern in the Canadian oil sands since they were identified as the main cause of acute toxicity of the wastewater stream following ore processing. Numerous analytical techniques are employed for NA measurement. A universally accepted method has yet to be developed. Additionally, all techniques are semiquantitative as appropriate standards for calibration do not exist. Properties such as toxicity, biodegradability, and susceptibility to other treatment regimes depend on NA structure; measurement of total NA concentration is insufficient when examining these properties in detail. The use of high-resolution methods reduces errors to which low-resolution techniques are prone, but are expensive and difficult to access. The protocols chosen for inclusion here are more widely accessible and sufficient for most applications. Total acid number and Fourier transform infrared spectroscopy are used to estimate acid content and acid-extractable organic content, respectively. Gas chromatography will specifically target NAs, and coupled with mass spectrometry, the characterization of an NA mixture is possible. However, while low-resolution gas chromatography–mass spectrometry is widely available and recommended for most uses, awareness that this technique is prone to misclassification and false positive data is essential.


Acid-extractable organics Fourier transform infrared spectroscopy Gas chromatography Mass spectrometry Naphthenic acids Oil sands process-affected water Solvent extraction Total acid number 


  1. 1.
    Brient JA, Wessner PJ, Doyle MN (2000) Naphthenic acids. In: Kirk-Othmer encyclopedia of chemical technology. Wiley, New YorkGoogle Scholar
  2. 2.
    Tissot BP, Welte DH (1984) Petroleum formation and occurrence, 2nd edn. Springer, BerlinCrossRefGoogle Scholar
  3. 3.
    Slavcheva E, Shone B, Turnbull A (1999) Review of naphthenic acid corrosion in oil refining. Br Corros J 34(2):125–131CrossRefGoogle Scholar
  4. 4.
    Allen EW (2008) Process water treatment in Canada’s oil sands industry: I. Target pollutants and treatment objectives. J Environ Eng Sci 7(2):123–138CrossRefGoogle Scholar
  5. 5.
    Headley JV, McMartin D (2004) A review of the occurrence and fate of naphthenic acids in aquatic environments. J Environ Sci Health A 39(8):1989–2010CrossRefGoogle Scholar
  6. 6.
    Barth T, Høiland S, Fotland P, Askvik KM, Pedersen BS, Borgund AE (2004) Acidic compounds in biodegraded petroleum. Org Geochem 35(11–12):1513–1525CrossRefGoogle Scholar
  7. 7.
    Misiti T, Tezel U, Pavlostathis SG (2013) Fate and effect of naphthenic acids on oil refinery activated sludge wastewater treatment systems. Water Res 47(1):449–460CrossRefPubMedGoogle Scholar
  8. 8.
    Masliyah J, Zhou ZJ, Xu Z, Czarnecki J, Hamza H (2004) Understanding water-based bitumen extraction from Athabasca oil sands. Can J Chem Eng 82(August):628–654Google Scholar
  9. 9.
    Young RF, Wismer WV, Fedorak PM (2008) Estimating naphthenic acids concentrations in laboratory-exposed fish and in fish from the wild. Chemosphere 73(4):498–505CrossRefPubMedGoogle Scholar
  10. 10.
    Clemente JS, Fedorak PM (2005) A review of the occurrence, analyses, toxicity, and biodegradation of naphthenic acids. Chemosphere 60(5):585–600CrossRefPubMedGoogle Scholar
  11. 11.
    Whitby C (2010) Microbial naphthenic acid degradation. Adv Appl Microbiol 70:93–125CrossRefPubMedGoogle Scholar
  12. 12.
    Zhao B, Currie R, Mian H (2012) Catalogue of analytical methods for naphthenic acids related to oil sands operations. Oil Sands Research and Information Network, University of Alberta, School of Energy and the Environment, Edmonton. OSRIN Report No. TR-21. 65 ppGoogle Scholar
  13. 13.
    Miskimmin B, Fedorak PM, Lauman R, Vinke K (2010) Oil sands water toxicity: a critical review. Natural Resources Canada, CanmetENERGY, DevonGoogle Scholar
  14. 14.
    Kindzierski W, Jin J, Gamal El-Din M (2012) Review of health effects of naphthenic acids: data gaps and implications for understanding human health risk. Oil Sands Research and Information Network, University of Alberta, School of Energy and the Environment, Edmonton. OSRIN Report No. TR-20. 43 ppGoogle Scholar
  15. 15.
    Holowenko FM, MacKinnon MD, Fedorak PM (2002) Characterization of naphthenic acids in oil sands wastewaters by gas chromatography–mass spectrometry. Water Res 36(11):2843–2855CrossRefPubMedGoogle Scholar
  16. 16.
    Martin JW, Han X, Headley JV (2008) Comparison of high- and low-resolution electrospray ionization mass spectrometry for the analysis of naphthenic acid mixtures in oil sands process water. Rapid Commun Mass Spectrom 22:1919–1924CrossRefPubMedGoogle Scholar
  17. 17.
    Headley JV, Peru KM, Mohamed MH et al (2013) Chemical fingerprinting of naphthenic acids and oil sands process waters: a review of analytical methods for environmental samples. J Environ Sci Health A Tox Hazard Subst Environ Eng 48(10):1145–1163CrossRefPubMedGoogle Scholar
  18. 18.
    Holowenko FM, MacKinnon MD, Fedorak PM (2001) Naphthenic acids and surrogate naphthenic acids in methanogenic microcosms. Water Res 35(11):2595–2606CrossRefPubMedGoogle Scholar
  19. 19.
    Scott AC, Young RF, Fedorak PM (2008) Comparison of GC–MS and FTIR methods for quantifying naphthenic acids in water samples. Chemosphere 73(8):1258–1264CrossRefPubMedGoogle Scholar
  20. 20.
    Grewer DM, Young RF, Whittal RM, Fedorak PM (2010) Naphthenic acids and other acid-extractables in water samples from Alberta: what is being measured? Sci Total Environ 408(23):5997–6010CrossRefPubMedGoogle Scholar
  21. 21.
    St. John WP, Rughani J, Green SA, Mcginnis GD (1998) Analysis and characterization of naphthenic acids by gas chromatography – electron impact mass spectrometry of tert-butyldimethylsilyl derivatives. J Chromatogr A 807:241–251CrossRefGoogle Scholar
  22. 22.
    Herman DC, Fedorak PM, MacKinnon MD, Costerton JW (1994) Biodegradation of naphthenic acids by microbial populations indigenous to oil sands tailings. Can J Microbiol 40(6):467–477CrossRefPubMedGoogle Scholar
  23. 23.
    Ross MS, Pereira ADS, Fennell J et al (2012) Quantitative and qualitative analysis of naphthenic acids in natural waters surrounding the Canadian oil sands industry. Environ Sci Technol 46(23):12796–12805CrossRefPubMedGoogle Scholar
  24. 24.
    Frank RA, Roy JW, Bickerton G et al (2014) Profiling oil sands mixtures from industrial developments and natural groundwaters for source identification. Environ Sci Technol 48(5):2660–2670CrossRefPubMedGoogle Scholar
  25. 25.
    Bataineh M, Scott AC, Fedorak PM, Martin JW (2006) Capillary HPLC/QTOF-MS for characterizing complex naphthenic acid mixtures and their microbial transformation. Anal Chem 78(24):8354–8361CrossRefPubMedGoogle Scholar
  26. 26.
    Hindle R, Noestheden M, Peru K, Headley J (2013) Quantitative analysis of naphthenic acids in water by liquid chromatography-accurate mass time-of-flight mass spectrometry. J Chromatogr A 1286:166–174CrossRefPubMedGoogle Scholar
  27. 27.
    Han X, MacKinnon MD, Martin JW (2009) Estimating the in situ biodegradation of naphthenic acids in oil sands process waters by HPLC/HRMS. Chemosphere 76(1):63–70CrossRefPubMedGoogle Scholar
  28. 28.
    Headley JV, Barrow MP, Peru KM et al (2011) Preliminary fingerprinting of Athabasca oil sands polar organics in environmental samples using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. Rapid Commun Mass Spectrom 25(13):1899–1909CrossRefPubMedGoogle Scholar
  29. 29.
    Pereira AS, Bhattacharjee S, Martin JW (2013) Characterization of oil sands process-affected waters by liquid chromatography orbitrap mass spectrometry. Environ Sci Technol 47(10):5504–5513CrossRefPubMedGoogle Scholar
  30. 30.
    Frank RA, Kavanagh R, Burnison BK et al (2006) Diethylaminoethyl-cellulose clean-up of a large volume naphthenic acid extract. Chemosphere 64(8):1346–1352CrossRefPubMedGoogle Scholar
  31. 31.
    Rowland SJ, Scarlett AG, Jones D, West CE, Frank RA (2011) Diamonds in the rough: identification of individual naphthenic acids in oil sands process water. Environ Sci Technol 45(7):3154–3159CrossRefPubMedGoogle Scholar
  32. 32.
    Jones D, Scarlett AG, West CE et al (2013) Elemental and spectroscopic characterization of fractions of an acidic extract of oil sands process water. Chemosphere 93(9):1655–1664CrossRefPubMedGoogle Scholar
  33. 33.
    Barrow MP, Headley JV, Derrick PJ (2009) Data visualization for the characterization of naphthenic acids within petroleum samples. Energy Fuel 23:2592–2599CrossRefGoogle Scholar
  34. 34.
    Headley JV, Peru KM, Fahlman B, Colodey A, McMartin DW (2013) Selective solvent extraction and characterization of the acid extractable fraction of Athabasca oils sands process waters by Orbitrap mass spectrometry. Int J Mass Spectrom 345–347:104–108CrossRefGoogle Scholar
  35. 35.
    Lu W, Ewanchuk A, Pérez-Estrada L, Sego D, Ulrich A (2013) Limitation of fluorescence spectrophotometry in the measurement of naphthenic acids in oil sands process water. J Environ Sci Health A Tox Hazard Subst Environ Eng 48(4):429–436CrossRefPubMedGoogle Scholar
  36. 36.
    Johnson RJ, Smith BE, Sutton PA, McGenity TJ, Rowland SJ, Whitby C (2011) Microbial biodegradation of aromatic alkanoic naphthenic acids is affected by the degree of alkyl side chain branching. ISME J 5(3):486–496CrossRefPubMedGoogle Scholar
  37. 37.
    Smith BE, Lewis CA, Belt ST, Whitby C, Rowland SJ (2008) Effects of alkyl chain branching on the biotransformation of naphthenic acids. Environ Sci Technol 42(24):9323–9328CrossRefPubMedGoogle Scholar
  38. 38.
    Fuhr B, Banjac B, Blackmore T, Rahimi P, Road KC, Tn C (2007) Applicability of total acid number analysis to heavy oils and bitumens. Energy Fuel 21:1322–1324CrossRefGoogle Scholar
  39. 39.
    Rogers VV, Liber K, MacKinnon MD (2002) Isolation and characterization of naphthenic acids from Athabasca oil sands tailings pond water. Chemosphere 48(5):519–527CrossRefPubMedGoogle Scholar
  40. 40.
    Headley JV, Peru KM, McMartin DW, Winkler M (2002) Determination of dissolved naphthenic acids in natural waters by using negative-ion electrospray mass spectrometry. J AOAC Int 85(1):182–187PubMedGoogle Scholar
  41. 41.
    Janfada A, Headley JV, Peru KM, Barbour SL (2006) A laboratory evaluation of the sorption of oil sands naphthenic acids on organic rich soils. J Environ Sci Health A Tox Hazard Subst Environ Eng 41(6):985–997CrossRefPubMedGoogle Scholar
  42. 42.
    Merlin M, Guigard SE, Fedorak PM (2007) Detecting naphthenic acids in waters by gas chromatography–mass spectrometry. J Chromatogr A 1140(1–2):225–229CrossRefPubMedGoogle Scholar
  43. 43.
    Headley JV, Peru KM, Barrow MP (2009) Mass spectrometric characterization of naphthenic acids in environmental samples: a review. Mass Spectrom Rev 28(1):121–134CrossRefPubMedGoogle Scholar
  44. 44.
    Herman DC, Fedorak PM, Costerton JW (1993) Biodegradation of cycloalkane carboxylic acids in oil sand tailings. Can J Microbiol 39(6):576–580CrossRefPubMedGoogle Scholar
  45. 45.
    Clemente JS, Fedorak PM (2004) Evaluation of the analyses of tert-butyldimethylsilyl derivatives of naphthenic acids by gas chromatography–electron impact mass spectrometry. J Chromatogr A 1047(1):117–128CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of Civil and Environmental Engineering3-091 Markin/CNRL Natural Resource Engineering Facility, University of AlbertaEdmontonCanada

Personalised recommendations