Development and Validation of an Ion Chromatography Method for the Simultaneous Determination of Seven Food Additives in Cheeses


Cheeses are characterized by several chemical-physical properties that make it difficult for the microorganisms growth, consequently. The actual European legislation allows the addition of few food additives in this type of food products. In this work, the entire procedure of extraction, purification, chromatographic separations and quali/quantitative determination of seven food additives (sorbic acid, benzoic acid, lactic acid, acetic acid, nitrites, nitrates and phosphates) was developed and applied for the analysis of different types of cheese (mozzarella, cheese spread, semi-hard and hard cheeses). Through validation procedure it was possible to evaluate the most important validation parameters. Extended calibration curves (r > 0.990) were obtained for all the analyzed compounds. Recovery values ranged from 72.8% to 98.4% and a good repeatability was obtained, with precision levels in the range of 0.03% - 0.11% (n = 6). The potential and feasibility of the method were tested by analysing real samples, such as mozzarella, cheese spread, semi-hard and hard cheeses, confirming that the method is well suited to satisfy the demands for accurate confirmation analyses of seven food additives in cheeses, which is especially valuable in official check analyses and in monitoring schemes.

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M. Iammarino and A. Taranto, "Development and Validation of an Ion Chromatography Method for the Simultaneous Determination of Seven Food Additives in Cheeses," Journal of Analytical Sciences, Methods and Instrumentation, Vol. 3 No. 3A, 2013, pp. 30-37. doi: 10.4236/jasmi.2013.33A005.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] P. F. Fox, P. L. H. Mc Sweeney and C. M. Lynch, “Significance of Non-Starter Lactic Acid Bacteria in Cheddar Cheese,” Australian Journal of Dairy Technology, Vol. 53, 1998, pp. 83-90.
[2] Commission Regulation, “Regulation (EC) No. 1129/ 2011 of 11 November 2011 Amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council by Establishing a Union List of Food Additives,” Official Journal of the European Communities, 2011, pp. L295/1-177.
[3] S. Bahruddin, B. Fazlul, I. S. Muhammad, A. Kamarudzaman and M. T. Khairuddin, “Simultaneous Determination of Preservatives (Benzoic Acid, Sorbic Acid, Methylparaben and Propylparaben) in Foodstuffs Using High-Performance Liquid Chromatography,” Journal of Chromatography A, Vol. 1073, No. 1-2, 2005, pp. 393-397.
[4] R. F. Witter, E. H. Newcomb and E. Stotz, “The Oxidation of Hexanoic Acid and Derivatives by Liver Tissue in Vitro,” The Journal of Biological Chemistry, Vol. 185, No. 2, 1950, pp. 537-548.
[5] H. J. Deuel, R. Alfin-Slater, C. S. Weil and H. F. Smyth, “Sorbic Acid as a Fungistatic Agent for Foods. I. Harmlessness of Sorbic Acid as a Dietary Component,” Food Research, Vol. 19, No. 1, 1954, p. 1.
[6] R. J. Feldmann and H. I. Maibach, “Absorption of Some Organic Compounds through the Skin in Man,” Journal of Investigative Dermatology Vol. 54, 1970, pp. 399-404.
[7] US FDA (United States Food and Drug Administration), “GRAS (Generally Recognized as Safe) Food Ingredients: Benzoic Acid and Sodium Benzoate,” Food and Drug Administration, Washington, DC, 1972.
[8] F. Feillet and J. V. Leonard, “Alternative Pathway Therapy for Urea Cycle Disorders,” Journal of Inherited Metabolic Disease, Vol. 21, No. 1, 1998, pp. 101-111.
[9] WHO (World Health Organization), “Concise International Chemical Assessment Document No. 26. Benzoic Acid and Sodium Benzoate,” Geneva, Switzerland, 2000.
[10] M. G. El-Ziney, “CG-MS Analysis of Benzoate and Sorbate in Saudi Dairy and Food Products with Estimation of Daily Exposure,” Journal of Food Technology, Vol. 7, No. 4, 2009, pp. 127-134.
[11] FAO/WHO (Food and Agriculture Organization of the United Nations/World Health Organization), “Toxicological Evaluation of Certain Food Additives,” 67th Meeting of the 435 Joint FAO/WHO Expert Committee on Food Additives (JECFA), 20-29 June 2006, Rome.
[12] M. Iammarino, A. Di Taranto, M. Muscarella and C. Palermo, “Survey of Benzoic Acid in Cheeses: Contribution to the Estimation of an Admissible Maximum Limit,” Food Additives and Contaminants: Part B, Vol. 4, No. 4, 2011, pp. 231-237.
[13] A. M. Fan and V. E. Steinberg, “Health Implications of Nitrate and Nitrite in Drinking Water: An Update on Methemoglobinemia Occurrence and Reproductive and Developmental Toxicity,” Regulatory Toxicology and Pharmacology, Vol. 23, No. 1, 1996, pp. 35-43.
[14] G. M. McKnight, C. W. Duncan, C. Leifert and M. H. Golden, “Dietary Nitrate in Man: Friend or Foe?” British Journal of Nutrition, Vol. 81, No. 5, 1999, pp. 349-358.
[15] O. Matteucci, G. Diletti, V. Principe, E. Di Giannatale, M. M. Marconi and G. Migliorati. “Due casi di Metaemoglobinemia Acuta da Sospetto Avvelenamento da Sodio Nitrito,” Veterinaria Italiana, Vol. 44, No. 2, 2008, pp. 439-445.
[16] M. C. Archer, “Mechanisms of Action of N-Nitroso Compounds,” Cancer Surveys, Vol. 8, 1989, pp. 241-250.
[17] Commission Regulation, “Regulation (EC) No. 1169/ 2011 of 25 October 2011 on the Provision of Food Information to Consumers, Amending Regulations (EC) No 1924/2006 and (EC) No 1925/2006 of the European Parliament and of the Council, and Repealing Commission Directive 87/250/EEC, Council Directive 90/496/EEC, Commission Directive 1999/10/EC, Directive 2000/13/ EC of the European Parliament and of the Council, Commission Directives 2002/67/EC and 2008/5/EC and Commission Regulation (EC) No 608/2004. Official Journal of the European Communities, 2011, pp. L304/ 18-63.
[18] M. S. Norma, J. Steinhardt Bour, M. S. Barbara, A. Soullier and M. B. Zemel, “Effect of Level and Form of Phosphorus and Level of Calcium Intake on Zinc, Iron and Copper Bioavailability in Man,” Nutrition Research, Vol. 4, No. 3, 1984, pp. 371-379.
[19] M. C. Carey, “Pathogenesis of Gallstones,” Recenti Progressi in Medicina, Vol. 83, No. 7-8, 1992, pp. 379-391.
[20] Commission Regulation, “Regulation (EC) No. 882/2004 of the European Parliament and of the Council of 29 April 2004,” Journal of the European Communities, 2004, pp. L165,1-141.
[21] Commission Decision, “Decision (EC) No. 657/2002 of 12 August 2002 Implementing Council Directive 96/23 /EC Concerning the Performance of Analytical Methods and the Interpretation of Results,” Journal of the European Communities, 2002, L221,8-36.
[22] E. J. C. Miller and J. N. Miller, “Statistics for Analytical Chemistry,” 3rd Edition, Ellis Horwood PTR Prentice Hall, New York, 1993, p. 115.
[23] W. J. Youden and E. H. Steiner, “Statistical Manual of the AOAC—Association of the Official Analytical Chemists,” AOAC-I, Washington DC, 1975.
[24] E. Hund, D. L. Massart and J. Smeyers-Verbeke, “Operational Definitions of Uncertainty” Trends Analytcal Chemistry, Vol. 20, No. 8, 2001, pp. 394-406.
[25] Dionex Corporation, “Product Manual for IonPac? AS11HC IonPac? AG11-HC,” Document No. 031333-07, Sunnyvale, 2008.
[26] F. Toldrà, “Handbook of Meat Processing,” John Wiley & Sons, Blackwell Publishing, New York, 2010.
[27] L. Brüggemann, W. Quapp and R. Wennrich, “Test for Non-Linearity Concerning Linear Calibrated Chemical Measurements,” Accreditation and Quality Assurance, Vol. 11, No. 12, 2006, pp. 625-631.
[28] M. Iammarino, A. Di Taranto and M. Cristino, “Endogenous Levels of Nitrites and Nitrates in Wide Consumption Foodstuffs: Results of Five Years of Official Controls and Monitoring,” Food Chemistry, Vol. 140, No. 15, 2013, pp. 763-771.
[29] M. Iammarino, A. Di Taranto, M. Muscarella and C. Palermo, “Assessment of Natural Levels of Substances with Preservative Effects in Dairy Products,” In: H. Castelli and L. du Vale, Eds., Handbook on Cheese, Production, Chemistry and Sensory Properties, Nova Publisher Inc., New York, 2013, pp. 559-571.
[30] L. J. Schuddeboom, “Nitrates and Nitrites in Foodstuffs,” Council of Europe, Strasbourg, 1993.
[31] A. Top?u, A. A. Top?u, I. Saldamli and M. Yurttagül, “Determination of Nitrate and Nitrite Content of Turkish Cheeses,” African Journal of Biotechnology, Vol. 5, No. 15, 2006, pp. 1411-1414.
[32] L. Tudor, E. Mitranescu, L. Tudor and F. Furnaris, “Assessment of Nitrate and Nitrite Content of Romanian Traditional Cheese,” Lucrari Stiintifice Medicina Veterinaria, Timisoara, XL, 2007.

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