Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-22T05:05:55.217Z Has data issue: false hasContentIssue false
  • English
  • Français

Direct estimation of sialic acid in milk and milk products by fluorimetry and its application in detection of sweet whey adulteration in milk

Published online by Cambridge University Press:  23 October 2012

Neelima ,
Priyanka Singh Rao ,
Rajan Sharma  and
Yudhishthir S. Rajput
Neelima
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132 001, India
Priyanka Singh Rao
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132 001, India
Rajan Sharma*
Affiliation:
Dairy Chemistry Division, National Dairy Research Institute, Karnal-132 001, India
Yudhishthir S. Rajput
Affiliation:
Division of Animal Biochemistry, Karnal-132 001, India
*
*For Correspondence; e-mail: rajansharma21@gmail.com
Get access Rights & Permissions [Opens in a new window]

Abstract

Sialic acid, being a biologically active compound, is recognised as an important component of milk and milk products. Almost all the sialic acid estimation protocols in milk require prior hydrolysis step to release the bound sialic acid followed by its estimation. The objective of this work was to estimate sialic acid in milk and milk products by fluorimetric assay which does not require a prior hydrolysis step thus decreasing the estimation time. The recovery of added sialic acid in milk was 91·6 to 95·8%. Sialic acid in milk was found to be dependent on cattle breed and was in the range of 1·68–3·93 g/kg (dry matter basis). The assay was further extended to detect adulteration of milk with sweet whey which is based on the detection of glycomacropeptide (GMP) bound sialic acid in adulterated milk. GMP is the C-terminal part of κ-casein which is released into the whey during cheese making. For detection of adulteration, selective precipitation of GMP was done using trichloroacetic acid (TCA). TCA concentration in milk was first raised to 5% to precipitate milk proteins, especially κ-casein, followed by raising the TCA concentration to 14% to precipitate out GMP. In the precipitates GMP bound sialic acid was estimated using fluorimetric method and the fluorescence intensity was found to be directly proportional to the level of sweet whey in adulterated milk samples. The method was found to detect the presence of 5% sweet whey in milk.

Keywords

Sialic acidFluorimetryMilkadulterationSweet wheyGlycomacropeptide
Type
Research Article
Information
Journal of Dairy Research , Volume 79 , Issue 4 , November 2012 , pp. 495 - 501
Copyright
Copyright © Proprietors of Journal of Dairy Research 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aminoff, D 1961 Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochemical Journal 81 384–392Google Scholar
Bremer, MGEG, Kemmers-Voncken, AEM, Boers, EAM, Frankhuizen, R & Haasnoot, W 2008 Enzyme-linked immunosorbent assay for the detection of rennet whey powder in milk and buttermilk powder. International Dairy Journal 18 294–302Google Scholar
Chavez, NA, Salinas, E, Jauregui, J, Palomares, LA & Macias, K 2008 Detection of bovine milk adulterated with cheese whey by western blot immunoassay. Food and Agricultural Immunology 19 265–272Google Scholar
Cherkaoui, S, Doumenc, N, Tachon, P, Neeser, JR & Veuthey, JL 1997 Development of a capillary zone electrophoresis method for caseinoglycomacropeptide determination. Journal of Chromatography A 790 195–205CrossRefGoogle ScholarPubMed
de Carvalho, BMA, de Carvalho, LM, Alcântra, LAP & Bonomo, RCF 2007 Métodos de detecção de fraude em leite por adição de soro de queijo. REDVET. Revista electrónica de Veterinaria 8 1–7Google Scholar
Fernandez, A, Menendez, V, Riera, FA & Alvarez, R 2011 Caseinomacropeptide behaviour in a whey protein fractionation process based on α-lactalbumin precipitation. Journal of Dairy Research 78 196–202Google Scholar
Fernando, SF & Woonton, BW 2010 Quantitation of N-acetylneuraminic (sialic) acid in bovine glycomacropeptide (GMP). Journal of Food Composition and Analysis 23 359–366CrossRefGoogle Scholar
Josephson, R, Holloway-Thomas, DJ & Warthesen, J 1980 Cheese whey detection in frozen desserts. Journal of Dairy Science 63 1356–1360CrossRefGoogle Scholar
Koliwer-Brandl, H, Siegert, N, Umnus, K, Kelm, A, Tolkach, A, Kulozik, U, Kuballa, J, Cartellieri, S & Kelm, S 2011 Lectin inhibition assays for the analysis of bioactive milk sialoglycoconjugates. International Dairy Journal 21 413–420Google Scholar
Koning, P, Elisses, J & De Vries, H 1966 A method for the detection of small percentages of whey powder in milk powder. Netherlands Milk and Dairy Journal 20 203–212Google Scholar
Lacomba, R, Salcedo, J, Alegria, A, Barbera, R, Hueso, P, Matencio, E & Lagarda, MJ 2011 Sialic acid (N-acetyl and N-glycolylneuraminic acid) and ganglioside in whey protein concentrates and infant formulae. International Dairy Journal 21 887–895CrossRefGoogle Scholar
Lacomba, R, Salcedo, J, Alegria, A, Lagarda, MJ, Barbera, R & Matencio, E 2010 Determination of sialic acid and gangliosides in biological samples and dairy products: A review. Journal of Pharmaceutical and Biomedical Analysis 51 346–357Google Scholar
Lynch, J 2005 Appendix E: laboratory quality assurance In Official Method of Analysis of AOAC International, (Eds Horwitz, W & Latimer, GW Jr). Geneva, Switzerland: AOAC InternationalGoogle Scholar
Martin, MJ, Vázquez, E & Rueda, R 2007 Application of a sensitive fluorometric HPLC assay to determine the sialic acid content of infant formulas. Analytical and Bioanalytical Chemistry 387 2943–2949Google Scholar
Martin-Hernandez, C, Munoz, M, Daury, C, Weymuth, H, Kemmers-Voncken, AEM, Corbation, V, Toribio, T & Bremer, MGEG 2009 Immunochromatographic lateral-flow test strip for the rapid detection of added bovine rennet whey in milk and milk powder. International Dairy Journal 19 205–208Google Scholar
Matsuno, K & Suzuki, S 2008 Simple fluorimetric method for quantification of sialic acids in glycoproteins. Analytical Biochemistry 375 53–59CrossRefGoogle ScholarPubMed
Medndham, J, Denney, RC, Barnes, JD & Thomas, MJK 2000 Vogels's Textbook of Quantitative Chemical Analysis. 6th edition. New Delhi, India: Pearson Education (Singapore) Pte. Ltd.Google Scholar
Nakano, T, Ikawa, N & Ozimek, L 2007 Detection of sialylated phosphorylated k-casein glycomacropeptide electrophoresed on polyacrylamide gels and cellulose acetate strips by the thiobarbituric acid and malachite green dye reactions. Journal of Agricultural and Food Chemistry 55 2714–2726Google Scholar
Nakano, T & Ozimek, L 1999 Determination of sialic acid by the thiobarbituric acid reaction in sweet whey and its fractions. Journal of Agriculture and Food Chemistry 47 2613–2616CrossRefGoogle ScholarPubMed
Romero, EL, Pardo, MF, Porro, S & Alonso, S 1997 Sialic acid measurement by a modified Aminoff method: a time-saving reduction in 2-thiobarbituric acid concentration. Journal of Biochemical and Biophysical Methods 35 129–134CrossRefGoogle Scholar
Saito, T, Yamaji, A & Itoh, T 1991 A new isolation method of caseinoglycopeptide from sweet cheese whey. Journal of Dairy Science 74 2831–2837CrossRefGoogle Scholar
Salcedo, J, Lacomba, R, Alegria, A, Barbera, R, Matencio, E & Lagarda, MJ 2011 Comparison of spectrophotometric and HPLC methods for determining sialic acid in infant formulas. Food Chemistry 127 1905–1910Google Scholar
Spichtig, V, Michaud, J & Austin, S 2010 Determination of sialic acids in milk and milk based products. Analytical Biochemistry 405 28–40Google Scholar
Sugahara, K, Sugimoto, K, Nomura, O & Usui, T 1980 Enzymatic assay of serum sialic acid. Clinica Chimica Acta 108 493–498Google Scholar
Teshima, S, Tamal, K, Hayashi, Y & Emi, S 1988 New enzymatic determination of sialic acid in serum. Clinical Chemistry 34 2291–2294Google Scholar
Thoma, C, Krause, I & Kulozik, U 2006 Precipitation behaviour of caseinomacropeptide and their simultaneous determination with whey proteins by RP-HPLC. International Dairy Journal 16 285–293Google Scholar
Thoma-Worringer, C, Sorensen, J & Lopez-Fandino, R 2006 Health effects and technological features of caseinomacropeptide. International Dairy Journal 16 1324–1333CrossRefGoogle Scholar
Wang, B & Brand-Miller, J 2003 The role and potential of sialic acid in human nutrition. European Journal of Clinical Nutrition 57 1351–1369Google Scholar
Wang, B, Brand-Miller, J, McVeagh, P & Petocz, P 2001 Concentration and distribution of sialic acid in human milk and infant formulas. American Journal of Clinical Nutrition 74 510–515CrossRefGoogle ScholarPubMed
Warren, L 1959 The thiobarbituric acid assay of sialic acids. Journal of Biological Chemistry 234 1971–1974Google Scholar