Chemometrics Analysis Combined with GC-MS and NMR Spectroscopy Analysis of Fatty Acids as a Means of Discriminating Butterfat Adulteration


  • Nurrulhidayah Ahmad Fadzillah Islamic University Malaysia, 25200 Kuantan, Malaysia
  • Abdul Rohman Gadjah Mada University, 55281 Yogyakarta, Indonesia
  • Amin Ismail Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
  • Yanty Noorziana Abdul Manaf Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
  • Arieff Salleh Rosman University Technology Malaysia, 81310 Johor Bahru, Johor Darul Takzim, Malaysia
  • Alfi Khatib Islamic University Malaysia, 25200 Kuantan, Malaysia
  • Norazian Mohd. Hassan Islamic University Malaysia, 25200 Kuantan, Malaysia
  • Rashidi Othman Islamic University Malaysia, 25200 Kuantan, Malaysia



Fatty acid, triglycerides, nutrition, adulteration, butter, lard, chemometrics.


The adulteration of butter has become a major problem in food industries. Butter has the similar characteristic to lard which makes lard a desirable adulterant in butter due to economic advantages. Therefore, the method of detection to analyse the adulteration practice must be developed. This study used NMR spectroscopy in combination with chemometrics for the authentication of butter from lard. The presence of lard as an adulterant in butter has been analysed using Gas Chromatography–Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy with the aid of chemometric of Principal Component Analysis (PCA) and Discriminant Analysis (DA). PC1 described 82% of the variation while PC2 accounted for 15% of the variation resulted in a model that described 80% of the total variance in the data. With 82% of the peak variation along the first PC, it was clear that all seventeen samples of butter in the market and pure lard sample were formed according to their own group and showed two well-defined and well-separated group. DA model classified 100% of all samples accurately according to its group (butter and butter adulterated with animal fats), meaning that no samples were misclassified into the wrong group. Lard was successfully determined at 2.63 ppm. In this study, NMR and PCA analysis has successfully discriminated between the market sample and lard and the results established that there is no lard being adulterated in all commercial butter samples. This could be a potential identification approach to determine if the product has been deceived in market.


Picariello G, Sacchi R, Fierro O, Melck D, Romano R, Paduano A. High resolution 13C NMR detection of short?and medium?chain synthetic triacylglycerols used in butterfat adulteration. Eur J Lipid Sci Technol 2013; 115: 858-864. DOI:

Jensen RG. The composition of bovine milk lipids: January 1995 to December 2000. J Dairy Sci 2002; 85(2): 295-350. DOI:

German JB, Gibson RA, Krauss RM, Nestle P, Lamarche B, van Staveren WA, Steijns JM, de Groot LCPGM, Lock AL, Destaillats F. A reappraisal of the impact of dairy foods and milk fat on cardiovascular disease risk. Eur J Nutr 2009; 48(4): 191-203. DOI:

Guyot A. Recent progress in reactive surfactants in emulsion polymerisation. Proceedings of the 2002 Macromolecular Symposia 2002; pp. 105-132.<105::AID-MASY105>3.0.CO;2-7 DOI:<105::AID-MASY105>3.0.CO;2-7

Barron LJ, Hierro MTG, Santa-María G. HPLC and GLC analysis of the triglyceride composition of bovine, ovine and caprine milk fat. J Dairy Sci 1990; 57(4): 517-526. DOI:

Lipp M. Review of methods for the analysis of triglycerides in milk fat: application for studies of milk quality and adulteration. Food Chem 1995; 54: 213-221. DOI:

Kuksis A, McCarthy MJ. Gas-liquid chromatographic fractionation of natural triglyceride mixtures by carbon number. Can J Biochem Physiol 1962; 40(5) :679-686. DOI:

Echizen A, Diki M. Determination of butterfat in mixed fats. Relation between butyric acid and butterfat content in mixed fats. Rep Cent Customs Lab 1975; 15:111.

Farag R, Abo-Raya S, Ahmed F, Hewedi F, Khalifa H. Fractional crystallization and gas chromatographic analysis of fatty acids as a means of detecting butterfat adulteration. J Am Oil Chem Soc 1983; 60: 1665-1669. DOI:

Iwaida M, lto Y, Tonogai Y, Toyoda M, Watanabe T. Detection of adulterated milk fats by gas-liquid chromatographic determination of butyrate and caproate, Food Hyg Safe Sci 1979; 20: 328. DOI:

Timms RE. Detection and quantification of non-milk fat in mixtures of milk and non-milk fats. J Dairy Res 1980; 47(3): 295-303. DOI:

Gaydou EM, Bianchini JP, Ratovohery JV. Triterpene alcohols, methylsterols, sterols, and fatty acids in five Malagasy legume seed oils. J Agric Food Chem 1983; 31(4): 833-836. DOI:

Harker M, Holmberg N, Clayton JC, Gibbard CL, Wallace AD, Rawlins S, Hellyer SA, Lanot A, Safford R. Enhancement of seed phytosterol levels by expression of an N?terminal truncated Hevea brasiliensis (rubber tree) 3?hydroxy?3?methylglutaryl?CoA reductase. Plant Biotechnol J 2003; 1(2): 113-121. DOI:

Holmberg K. Natural surfactants. Curr Opin Colloid Interface Sci 2001; 6:148-159. DOI:

Hubbard WD, Sheppard AJ, Newkirk DR, Prosser AR, Osgood T. Comparison of various methods for the extraction of total lipids, fatty acids, cholesterol, and other sterols from food products. J Am Oil Chem Soc 1977; 54(2): 81-83. DOI:

Freire E. Differential scanning calorimetry. Protein Stability and Folding: Theory and Practice. Methods Mol Biol 1995; 40: 191-218 DOI:

Kittur F, Prashanth KH, Sankar KU, Tharanathan R. Characterization of chitin, chitosan and their carboxymethyl derivatives by differential scanning calorimetry. Carbohydr Polym 2002; 49: 185-193. DOI:

Lees M. Food authenticity and traceability. 1st ed. USA: Woodhead Publishing 2003. DOI:

Rohman A, Sismindari Erwanto Y, Che Man YB. Analysis of pork adulteration in beef meatball using Fourier transform infrared (FTIR) spectroscopy. Meat Sci 2011; 88(1): 91-95. DOI:

Christy AA, Egeberg PK. Quantitative determination of saturated and unsaturated fatty acids in edible oils by infrared spectroscopy and chemometrics. Chemom Intell Lab Syst 2006; 82:130-136. DOI:

Shin EC, Craft BD, Peg RB, Phillips RD, Eitenmiller RR. Chemometric approach to fatty acid profiles in Runner-type peanut cultivars by principal component analysis (PCA). Food Chem 2010; 119(3): 1262-1270. DOI:

Rohman A, Che Man YB. Analysis of cod-liver oil adulteration using Fourier transform infrared (FTIR) spectroscopy. J Am Oil Chem Soc 2009; 86: 1149-1153. DOI:

AOAC (Association of Official Analytical Chemists). Official methods of analysis of AOAC International (Official Method 920.118). 17th ed. Gaithersbug: MD 2000.

De Pedro E, Casillas M, Miranda C. Microwave Oven Application in the Extraction of Fat from the Subcutaneous Tissue of Iberian Pig Ham. Meat Sci 1997; 45: 45-51. DOI:




How to Cite

Nurrulhidayah Ahmad Fadzillah, Abdul Rohman, Amin Ismail, Yanty Noorziana Abdul Manaf, Arieff Salleh Rosman, Alfi Khatib, Norazian Mohd. Hassan, & Rashidi Othman. (2019). Chemometrics Analysis Combined with GC-MS and NMR Spectroscopy Analysis of Fatty Acids as a Means of Discriminating Butterfat Adulteration . Journal of Pharmacy and Nutrition Sciences, 9(2), 87–94.