Skip to main content
Log in

Application of Silver Ion High-Performance Liquid Chromatography for Quantitative Analysis of Selected n-3 and n-6 PUFA in Oil Supplements

  • Original Article
  • Published:
Lipids

Abstract

The aim of this study was to develop a simple method for simultaneous determination of selected cis/cis PUFA–LNA (18:2), ALA (18:3), GLA (18:3), EPA (20:5), and DHA (22:6) by silver ion high-performance liquid chromatography coupled to a diode array detector (Ag-HPLC-DAD). The separation was performed on three Luna SCX Silver Loaded columns connected in series maintained at 10 °C with isocratic elution by 1 % acetonitrile in n-hexane. The applied chromatographic system allowed a baseline separation of standard mixture of n-3 and n-6 fatty acid methyl esters containing LNA, DHA, and EPA and partial separation of ALA and GLA positional isomers. The method was validated by means of linearity, precision, stability, and recovery. Limits of detection (LOD) for considered PUFA standard solutions ranged from 0.27 to 0.43 mg L−1. The developed method was used to evaluate of n-3 and n-6 fatty acids contents in plant and fish softgel oil capsules, results were compared with reference GC-FID based method.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Calder PC, Yaqoob P (2009) Omega-3 polyunsaturated fatty acids and human health outcomes. Biofactors 35:266–272

    Article  CAS  PubMed  Google Scholar 

  2. Dunbar BS, Bosire RV, Deckelbaum RJ (2014) Omega 3 and omega 6 fatty acids in human and animal health: an African perspective. Mol Cell Endocrinol 398:69–77

    Article  CAS  PubMed  Google Scholar 

  3. González-Rodríguez LG, Aparicio A, López-Sobaler AM, Ortega RM (2013) Omega 3 and omega 6 fatty acids intake and dietary sources in a representative sample of Spanish adults. Int J Vitam Nutr Res 83:36–47

    Article  PubMed  Google Scholar 

  4. Watters C, Edmonds C, Rosner L, Sloss K, Leung P (2012) A cost analysis of EPA and DHA in fish, supplements and foods. J Nutr Food Sci. doi:10.4172/2155-9600.1000159

    Google Scholar 

  5. Kleiner AC, Cladis DP, Santerre CR (2015) A comparison of actual versus stated label amounts of EPA and DHA in commercial omega-3 dietary supplements in the United States. J Sci Food Agric 95:1260–1267

    Article  CAS  PubMed  Google Scholar 

  6. Villegas C, Zhao Y, Curtis JM (2010) Two methods for the separation of monounsaturated octadecenoic acid isomers. J Chromatogr A 1217:775–784

    Article  CAS  PubMed  Google Scholar 

  7. Jensen RG (2002) The composition of bovine milk lipids: January 1995 to December 2000. J Dairy Sci 85:295–350

    Article  CAS  PubMed  Google Scholar 

  8. Nikolova-Damyanova B (2009) Retention of lipids in silver ion high-performance liquid chromatography: facts and assumptions. J Chromatogr A 1216:1815–1824

    Article  CAS  PubMed  Google Scholar 

  9. Dewar MJS (1951) Bull Soc Chim France 18:C71

    Google Scholar 

  10. Frenking G, Fröhlich N (2000) The nature of the bonding in transition-metal compounds. Chem Rev 100:717–774

    Article  CAS  PubMed  Google Scholar 

  11. Momchilova SM, Nikolova-Damyanova BM (2012) Advances in silver ion chromatography for the analysis of fatty acids and triacylglycerols—2001 to 2011. Anal Sci 28:837–844

    Article  CAS  PubMed  Google Scholar 

  12. Dillon JT, Aponte JC, Tarozo R, Huang Y (2013) Purification of omega-3 polyunsaturated fatty acids from fish oil using silver-thiolate chromatographic material and high performance liquid chromatography. J Chromatogr A 1312:18–25

    Article  CAS  PubMed  Google Scholar 

  13. Adlof RO (1997) Normal-phase separation effects with lipids on a silver ion high-performance liquid chromatography column. J Chromatogr A 764:337–340

    Article  CAS  Google Scholar 

  14. Dreiucker J, Vetter W (2011) Fatty acids patterns in camel, moose, cow and human milk as determined with GC/MS after silver ion solid phase extraction. Food Chem 126:762–771

    Article  CAS  Google Scholar 

  15. Christie WW, Dobson G, Adlof RO (2007) A practical guide to the isolation, analysis and identification of conjugated linoleic acid. Lipids 42:1073–1084

    Article  CAS  PubMed  Google Scholar 

  16. Ostrowska E, Dunshes FR, Muralitharan M, Cross RF (2000) Comparison of silver-ion high-performance liquid chromatographic quantification of free and methylated conjugated linoleic acids. Lipids 35:1147–1153

    Article  CAS  PubMed  Google Scholar 

  17. Czauderna M, Kowalczyk J, Marounek M, Michalski JP, Rozbicka-Wieczorek AJ, Krajewska KA (2011) A new internal standard for HPLC assay of conjugated linoleic acid in animal tissues and milk. Czech J Anim Sci 56:23–29

    CAS  Google Scholar 

  18. Rodríguez-Castañedas JL, Peña-Egido MJ, García-Marino M, García-Moreno C (2011) Quantitative determination of conjugated linoleic acid isomers by silver ion HPLC in ewe milk fat. J Food Comp Anal 24:1004–1008

    Article  Google Scholar 

  19. Nikolova-Damyanova B, Momchilova S, Christie WW (2000) Silver ion high-performance liquid chromatographic separation of conjugated linoleic acid isomers, and other fatty acids, after conversion to p-methoxyphenacyl derivatives. J High Resol Chromatogr 23:348–352

    Article  CAS  Google Scholar 

  20. Cross RF, Ostrowska E (2000) Mixed mode retention and the use of competing acid for the Ag + -HPLC analysis of underivatized conjugated linoleic acids. J High Resol Chromatogr 23:317–323

    Article  CAS  Google Scholar 

  21. Adlof RO (1994) Separation of cis and trans unsaturated fatty acid methyl esters by silver ion high-performance liquid chromatography. J Chromatogr A 659:95–99

    Article  CAS  Google Scholar 

  22. Gunstone FD, Ismail IA, Lie Ken Jie M (1967) Fatty acids, part 16. Thin layer and gas–liquid chromatographic properties of the cis and trans methyl octadecenoates and of some acetylenic esters. Chem Phys Lipids 1:376–386

    Article  CAS  Google Scholar 

  23. Gunstone FD, Lie Ken Jie M (1970) Fatty acids, part 26. Thin layer and gas liquid chromatographic properties of some cis-octadecenyl compounds. Chem Phys Lipids 4:139–146

    Article  CAS  Google Scholar 

  24. Nikolova-Damyanova B, Herslof B, Christie WW (1992) Silver ion high-performance liquid chromatography of derivatives of isomeric fatty acids. J Chromatogr A 609:133–140

    Article  CAS  Google Scholar 

  25. Nikolova-Damyanova B, Christie WW, Herslof B (1995) Silver ion high-performance liquid chromatography of esters of isomeric octadecenoic fatty acids with short-chain monounsaturated alcohols. J Chromatogr A 693:235–239

    Article  CAS  Google Scholar 

  26. Nikolova-Damyanova B, Christie WW, Herslof B (1996) Mechanistic aspects of fatty acid retention in silver ion chromatography. J Chromatogr A 749:47–54

    Article  CAS  Google Scholar 

  27. Fardin-Kia AR, Delmonte P, Kramer JKG, Jahreis G, Kuhnt K, Santercole V, Rader JI (2013) Separation of the fatty acids in menhaden oil as methyl esters with a highly polar ionic liquid gas chromatographic column and identification by time of flight mass spectrometry. Lipids 48:1279–1295

    Article  CAS  PubMed  Google Scholar 

  28. Adlof RO (2007) Analysis of triacylglycerol and fatty acid isomers by low-temperature silver-ion high performance liquid chromatography with acetonitrile in hexane as solvent: limitations of the methodology. J Chromatogr A 1148:256–259

    Article  CAS  PubMed  Google Scholar 

  29. Adlof RO, List G (2004) Analysis of triglyceride isomers by silver-ion high-performance liquid chromatography: effect of column temperature on retention times. J Chromatogr A 1046:109–113

    Article  CAS  PubMed  Google Scholar 

  30. De La Fuente MA, Luna P, Juarez M (2006) Chromatographic techniques to determine conjugated linoleic acid isomers. Trends in Anal Chem 25:917–926

    Article  Google Scholar 

  31. Rodriguez-Alcala LM, Fontecha J (2007) Hot topic: Fatty acid and conjugated linoleic acid (CLA) isomer composition of commercial CLA-fortified dairy products: Evaluation after processing and storage. J Dairy Sci 90:2083–2090

    Article  CAS  PubMed  Google Scholar 

  32. Nunes JC, Torres AG (2010) Fatty acid and CLA composition of Brazilian dairy products, and contribution to daily intake of CLA. J Food Comp Anal 23:782–789

    Article  CAS  Google Scholar 

  33. Gomez-Cortes P, Frutos P, Mantecon AR, Juarez M, De La Fuente MA, Hervas G (2008) Addition of olive oil to dairy ewe diets: effect on milk fatty acid profile and animal performance. J Dairy Sci 91:3119–3127

    Article  CAS  PubMed  Google Scholar 

  34. Luna P, Juarez M, De La Fuente MA (2007) Fatty acid and conjugated linoleic acid isomer profiles in human milk fat. Eur J Lipid Sci Technol 109:1160–1166

    Article  CAS  Google Scholar 

  35. Nuernberg K, Dannenberger D, Ender K, Nuernberg G (2007) Comparison of different methylation methods for the analysis of conjugated linoleic acid isomers by silver ion HPLC in beef lipids. J Agric Food Chem 55:598–602

    Article  CAS  PubMed  Google Scholar 

  36. Toral PG, Shingfield KJ, Hervas G, Toivonen V, Frutos P (2010) Effect of fish oil and sunflower oil on rumen fermentation characteristics and fatty acid composition of digesta in ewes fed a high concentrate diet. J Dairy Sci 93:4804–4817

    Article  CAS  PubMed  Google Scholar 

  37. Rego OA, Alves SP, Antunes LMS, Rosa HJD, Alfaia CFM, Prates JAM, Cabrita ARJ, Fonseca AJM, Bessa RJB (2009) Rumen biohydrogenation-derived fatty acids in milk fat from grazing dairy cows supplemented with rapeseed, sunflower, or linseed oils. J Dairy Sci 92:4530–4540

    Article  CAS  PubMed  Google Scholar 

  38. Cruz-Hernandez C, Deng Z, Zhou J, Hill AR, Yurawecz MP, Delmonte P, Mossoba MM, Dugan MER, Michael ER, Kramer JKG (2004) Methods for analysis of conjugated linoleic acids and trans-18:1 isomers in dairy fats by using a combination of gas chromatography, silver-ion thin-layer chromatography/gas chromatography, and silver-ion liquid chromatography. J AOAC Int 87:545–562

    CAS  PubMed  Google Scholar 

  39. Juaneda P, Ledoux M, Sebedio JL (2007) Analytical methods for determination of trans fatty acid content in food. Eur J Lipid Sci Technol 109:901–917

    Article  CAS  Google Scholar 

  40. Fournier V, Juaneda P, Destaillats F, Dionisi F, Lambelet P, Sebedio JL, Berdeaqux O (2006) Analysis of eicosapentaenoic and docosahexaenoic acid geometrical isomers formed during fish oil deodorization. J Chromatogr A 1129:21–28

    Article  CAS  PubMed  Google Scholar 

  41. Delmonte P, Fardin-Kia AR, Kramer JKG, Mossoba MM (2011) Separation characteristics of fatty acid methyl esters using SLB-IL111, a new ionic liquid coated capillary gas chromatographic column. J Chromatogr A 1218:545–554

    Article  CAS  PubMed  Google Scholar 

  42. Delmonte P, Fardin-Kia AR, Kramer JKG, Mossoba MM, Sidisky L, Tyburczy C, Rader JI (2012) Evaluation of highly polar ionic liquid gas chromatographic column for the determination of the fatty acids in milk fat. J Chromatogr A 1233:137–146

    Article  CAS  PubMed  Google Scholar 

  43. Marekov I, Panayotova S, Tarandjiiska R (2009) Fatty acid composition and seasonal variation of trans fatty acid content in Bulgarian butter fats by silver ion TLC and GC. J Liq Chromatogr Relat Technol 32:1183–1195

    Article  CAS  Google Scholar 

  44. Marekov I, Nedelcheva D, Panayotova S, Tarandjiiska R (2011) Detection of milkfat adulteration by GC analysis of saturated, cis-monoenoic and cis, cis-dienoic fatty acid fractions isolated by silver ion TLC. J Liq Chromatogr Relat Technol 34:888–901

    Article  CAS  Google Scholar 

  45. Marekov I, Tarandjiiska R, Panayotova S, Nikolova N (2002) Comparison of fatty acid composition of domestic and imported margarines and frying fats in Bulgaria. Eur J Lipid Sci. Technol 104:410–418

    Article  CAS  Google Scholar 

  46. Denev RV, Kuzmanova IS, Momchilova SM, Nikolova-Damyanova BM (2011) Resolution and quantification of isomeric fatty acids by silver ion HPLC: fatty acid composition of aniseed oil (Pimpinella anisum, Apiaceae). J AOAC Int 94:4–8

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The first author would like to thank Dr. Marek Roszko for his valuable editing assistance of this manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Anna Czajkowska-Mysłek.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Czajkowska-Mysłek, A., Siekierko, U. & Gajewska, M. Application of Silver Ion High-Performance Liquid Chromatography for Quantitative Analysis of Selected n-3 and n-6 PUFA in Oil Supplements. Lipids 51, 413–421 (2016). https://doi.org/10.1007/s11745-016-4133-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-016-4133-1

Keywords

Navigation