AJAC  Vol.3 No.4 , April 2012
Comparative Analysis and Validation Methodologies of GC and HPLC for Analysis of Cholesterol in Meat Products
Abstract: This study validated different extraction methodologies and compared the quantification of cholesterol by gas chromatography (GC) and high performance liquid chromatography (HPLC) in mg per 100 g of Italian-type salami and traditional bologna. The GC method used was direct saponification of the samples without derivatizations and the HPLC method was used to extract of the lipid samples. The GC limits of detection and quantification obtained for cholesterol were, respectively, 0.001 and 0.003 mg.g–1. The HPLC values were 0.005 mg.g–1 and 0.016 mg.g–1. The GC recovery rate was 97.10 ± 0.13 and that of HPLC was 93.33 ± 0.22. Comparison of the cholesterol quantity found using the two chromatographic techniques shows that both are capable of quantifying cholesterol in the foods. With regard to costs, analysis time, the cost/benefit relationship was better with gas chromatography than that obtained with high performance liquid chromatography.
Cite this paper: G. Luzia Stroher, A. Claudia Rodrigues, L. Felicidade Dias, M. Reghiany Pedrão, L. Nascimento de Paula, J. Vergilio Visentainer and N. Evelazio de Souza, "Comparative Analysis and Validation Methodologies of GC and HPLC for Analysis of Cholesterol in Meat Products," American Journal of Analytical Chemistry, Vol. 3 No. 4, 2012, pp. 306-311. doi: 10.4236/ajac.2012.34042.

[1]   M. Careri, A. Mangia, M. Musci and G. Parolari, “Development of Gas Chromatographic Method for Determination of Cholesterol Oxides in Meat Products,” Chromatographia, Vol. 47, 1998, pp. 42-48.

[2]   Ministry of Health, “Manual Treatment of Hereditary Co-agulation Disorders (Series A. Technical Standards and Manuals),” Ministry of Health, Brasília, 2006.

[3]   N. Bragagnolo and D. B. Rodriguez-Amaya, “Teores de Colesterol, Lipídios Totais e ácidos Graxos em Cortes de Carne Suína,” Ciência e Tecnologia de Alimentos, Vol. 22, No. 1, 2002, pp. 98-104. doi:10.1590/S0101-20612002000100018

[4]   N. M. W. Pratiwi, P. J. Murray and D, G. Taylor, “Total Cholesterol Concentrations of the Muscles in Castrated Boer Goats,” Small Ruminant Research, Vol. 64, No. 1-2, 2006, pp. 77-81. doi:10.1016/j.smallrumres.2005.04.016

[5]   J. Dong, W. Chen, S. Wang, J. Zhang, H. Li, H., Guo, Y. Man and B. Chen, “Jones Oxidation and High Performance Liquid Chromatographic Analysis of Cholesterol in Biological Samples,” Journal of Chromatography B, Vol. 858, No. 1-2, 2007, pp. 239-246. doi:10.1016/j.jchromb.2007.08.038

[6]   T. T. N. Dinh, J. R. Blanton Jr., J. C. Brooks, M. F. Miller and L. D. Thompson, “A Simplified Method for Choles-terol Determination in Meat and Meat Products,” Journal of Food Composition and Analysis, Vol. 21, No. 4, 2008, pp. 306-314. doi:10.1016/j.jfca.2008.02.001

[7]   A. Daneshfar, T. Khezeli and H. J. Lotfi, “Determination of Cholesterol in Food Samples Using Dispersive Liquid-Liquid Microextraction Followed by HPLC-UV,” Journal of Chromatography B, Vol. 877, No. 4, 2009, pp. 456-460. doi:10.1016/j.jchromb.2008.12.050

[8]   H. Farwanah, J. Wirtz, T. Kolter, K. Raith, R. H. H. Neu- bert and K. Sandhoff, “Normal Phase Liquid Chromatography Coupled to Quadrupole Time of Flight Atmospheric Pressure Chemical Ionization Mass Spectrometry for Separation, Detection and Mass Spectrometric Profiling of Neutral Sphingolipids and Cholesterol,” Journal of Chromatogry B, Vol. 877, No. 27, 2009, pp. 2976-2982. doi:10.1016/j.jchromb.2009.07.008

[9]   W. J. Griffiths and Y. Wang, “Analysis of Neurosterols by GC-MS and LC-MS/MS,” Journal of Chromatography B, Vol. 877, No. 26, 2009, pp. 2778-2805. doi:10.1016/j.jchromb.2009.05.017

[10]   V. A. Isidorov and L. Szczepaniak, “Gas Chromatographic Retention Indices of Biologically and Environmentally Important Organic Compounds on Capillary Col- umns with Low-Polar Stationary Phases,” Journal of Chromatography A, Vol. 1216, No. 51, 2009, pp. 8998- 9007. doi:10.1016/j.chroma.2009.10.079

[11]   S. M. Al-Hasani, J. Hlavac and M. W. Carpenter, “Rapid Determination of Cholesterol in Single and Multicomponent Prepared Foods,” Journal of AOAC International, Vol. 76, No. 4, 1993, pp. 902-906.

[12]   N. Bragagnolo, T. Saldanha and M. R. Mazalli, “Ava- lia??o Comparativa Entre dois Métodos Para Determi- na??o do Colesterol em Carnes e Leite,” Ciência e Tecnologia de Alimentos, Vol. 24, No. 1, 2004, pp. 109-113. doi:10.1590/S0101-20612004000100013

[13]   F. Dionisi, P. A. Golay, J. M. Aeschlimann and L. B. Fay, “Determination of Cholesterol Oxidation Products in Milk Powder: Methods Comparison and Validation,” Journal of Agricultural and Food Chemistry, Vol. 46, No. 6, 1998, pp. 2227-2233. doi:10.1021/jf971060

[14]   J. Toivo, K. Phillips, A. M. Lampi and V. Piironen, “Determination of Sterols in Foods: Recovery of Free, Esterified, Glycosidic Sterols,” Journal of Food Composition and Analysis, Vol. 14, No. 6, 2001, pp. 631-643. doi:10.1006/jfca.2001.1019

[15]   E. G. Bligh and E. J. Dyer, “A Rapid Method of Total Lipid Extraction and Purification,” Canadian Journal of Biochemistry and Physiology, Vol. 37, No. 8, 1959, pp. 911-917. doi:10.1139/o59-099

[16]   A. F. Verga, N. M. Saito, Y. S. Hirata and C. Barros, “Valida??o de Métodos Analíticos Para Fins de Certifica??o de Laboratórios,” Expo-Labor, Vol. 15, No. 3, 2009, pp. 115-128.

[17]   M. A. Campanero, “Development and Validation of a HPLC Method for Determination of Amphotericin B in Plasma and Sputum Involving Solid Phase Extraction,” Chromatographia, Vol. 46, No. 11-12, 1997, pp. 641-647. doi:10.1007/BF02490525

[18]   C. M. Bandeira, J. M. Ferreira, N. Bragagnolo and L. R. B. Mariutti, “Desenvolvimento de Metodologia Analítica Para Determina??o de Colesterol em Ra??o Para Rumi- nantes Através de Planejamento Experimental Fatorial,” Química Nova, Vol. 31, No. 6, 2008, pp. 1422-1426. doi:10.1590/S0100-40422008000600027

[19]   S. L. Nixdorf, M. Matsushita and N. E. Souza, “Coles- terol em Refei??es Comerciais Consumidas na Cidade de Londrina (PR), Brasil,” Boletim CEPPA, Vol. 15, 1997, pp. 149-158.

[20]   Ministry of Agriculture, Livestock and Supply, “Annex II. Technical Regulations for Food Labeling,” Official Gazette, Instruction No. 4, 31 March 2000.

[21]   F. Jimenez-Colmenero, J. Carballo and S. Cofrades, “Healthier Meat and Meat Products: Their Role as Functional Foods,” Meat Science, Vol. 59, No. 1, 2001, pp. 5-13. doi:10.1016/S0309-1740(01)00053-5

[22]   J. D. Beyer, F. X. Milani, M. J. Dutelle and R. L. Bradley Jr., “Gas Chromatographic Determination of Cholesterol in Egg Products,” Journal of Association of Official Analytical Chemists, Vol. 72, No. 5, 1989, pp. 746-748.

[23]   J. Folch, M. Less and S. Stanley, “A Simple Method for the Isolation and Purification of Total Lipids from Animal Tissues,” Journal of Biological Chemistry, Vol. 226, No. 1, 1957, pp. 497-509.

[24]   M. Fenton, “Review: Chromatographic Separation of Cholesterol in Foods,” Journal of Chromatography, Vol. 624, No. 1-2, 1992, pp. 369-388. doi:10.1016/0021-9673(92)85689-Q

[25]   M. Fenton and J. S. Sim, “Determination of Egg Yolk Cholesterol Content by On-Column Capillary Gás Chromatography,” Journal of Chromatography, Vol. 540, 1991, pp. 323-329. doi:10.1016/S0021-9673(01)88819-1

[26]   L. Alonso, L. Lozada, J. Fontecha and M. Juárez, “Determination of Cholesterol in Milk Fat by Gas Chromatography with Direct Injection and Sample Saponification,” Chromatographia, Vol. 41, No. 1-2, 1995, pp. 23- 28. doi:10.1007/BF02274190

[27]   United States Department of Agriculture, Agricultural Re- search Service, 2006. USDA Nutrient Database for Stan- dart Reference (Release 15). Nutriente Data Laboratory Home Page, 2006.

[28]   G. Lercker and M. T. Rodriguez-Estrada, “Cholesterol Oxidation: Presence of 7-Ketocholesterol in Different Food Products,” Journal of Food Composition and Analysis, Vol. 13, No. 4, 2000, pp. 625-631. doi:10.1006/jfca.2000.0901

[29]   S. R. Baggio and N. Bragagnolo, “Forma??o de óxidos de Colesterol e Altera??es dos ácidos Graxos em Produtos Cárneos,” Revista do Instituto Adolfo Lutz, Vol. 66, 2007, pp. 10-17.

[30]   ANVISA, “National Agency of Sanitary Surveillance,” Official Gazette, RDC-Resolution No. 360, 23 December 2003.

[31]   C. Maraschielo, I. Diaz and J. A. G. Regueiro, “Determination of Cholesterol in Fat and Muscle of Pig by HPLC and Capillary Gás Chromatography with Solvent Venting Injection,” Journal of High Resolution Chromatography, Vol. 19, No. 3, 1996, pp. 165-168.

[32]   E. Novelli, E. Zanardi, G. P. Ghiretti, G. Campanini, G. Dazzi, G. Madarena and R. Chizzolini, “Lipid and Cho- lesterol Oxidation in Frozen Stored Pork, Salame Milano and Mortadella,” Meat Science, Vol. 48, No. 1-2, 1998, pp. 29-40. doi:10.1016/S0309-1740(97)00072-7

[33]   Food and Drug Administration, “Food Labeling: Nutrient Content Claims, General Principles, Petitions, Definition of Terms; Definitions of Nutrient Content Claims for the Fat, Fatty Acid, Cholesterol Content of Food,” Federal Register, Vol. 58, No. 3, pp. 2302-2426.

[34]   M. R. Mazalli, A. C. H. F. Sawaya, M. N. Eberlin and N. Bragagnolo, “HPLC Method for Quantification and Characterization of Cholesterol and Its Oxidation Products in Eggs,” Lipids, Vol. 41, No. 6, 2006, pp. 615-622. doi:10.1007/s11745-006-5010-0

[35]   K. B. Choudhari, S. Jayanthi, R. B. Murty and R. R. Matharu, “A High-Performance Liquid Chromatographic Method for the Analysis of Lipids from Lyophilized Formulations,” Journal of Chromatography A, Vol. 724, No. 1-2, 1996, pp. 343-347. doi:10.1016/0021-9673(95)00933-7