PENGEMBANGAN PROSES SAPONIFIKASI DAN KONDISI REVERSED PHASE-HPLC-MWD PADA ANALISIS KOLESTEROL TELUR DARI METODE STANDAR AOAC 994.10:2012

Didah Nur Faridah, Yulina Lailatul Maslukhah, Ririn Anggraeni, Hanifah Nuryani Lioe

Abstract

Penentuan komposisi fase gerak, efisiensi waktu analisis, kromatogram yang bagus dan kehandalan instrumen perlu dipertimbangkan sebelum pengembangan metode analisis kolesterol menggunakan Reversed Phase-HPLC-Multiwavelength Detector (MWD). Konsentrasi KOH dan waktu saponifikasi diketahui berpengaruh terhadap kadar kolesterol yang diperoleh. Tujuan penelitian ini yaitu untuk mengembangkan metode kondisi RP-HPLC-MWD sebelum digunakan untuk analisis kolesterol, menguji kehandalan instrumennya, serta mengembangkan metode analisis kolesterol dalam telur ayam broiler dengan mengkombinasikan konsentrasi KOH dan waktu saponifikasi, pada suhu 80°C. Kombinasi acetonitril:metanol sebagai fase gerak (80:20, 20:80, 10:90, 0:100 v/v) dengan laju alir 1.2 mL/menit dan fase gerak 0:100 dengan laju alir 1.0 dan 0.8 mL/menit pada pada λ= 200, 205, 210 nm. Konsentrasi KOH dan waktu saponifikasi yang digunakan untuk pengembangan metode analisis kolesterol telur ayam yaitu 5 dan 10 % selama 15, 30, 45 menit. Kondisi RP-HPLC-MWD yang terpilih yaitu komposisi fase gerak metanol 100%, dengan laju alir fase gerak 1.0 mL/menit, λ= 205 nm. Instrumen ini mempunyai linieritas yang bagus, R2 = 0.9991. Limit deteksi dan kuantifikasi instrumen ini secara berurutan yaitu 5 dan 10 μg/mL. Konsentrasi KOH sebesar 10% (b/v) dan waktu saponifikasi selama 15 menit menghasilkan kadar kolesterol tertinggi dengan presisi yang bagus.

Keywords

detector multiwavelength, kehandalan instrumen, kriteria kromatografi.

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References

Acimovic J, Rozman D. (2013). Steroidal Triterpenes of Cholesterol Synthesis. Molecules. 18: 4002-4017. doi:10.3390/molecules18044002.Ahn, J.H., Jeong, I.S.,

Kwak, B.M., Leem, D., Yoon, T., Yoon, C., Jeong, J., Park, J.M., Kim, J.M. (2012). Rapid Determination of Cholesterol in Milk Containing Emulsified Foods. Food Chem, 135, 2411–2417. doi:10.1016/j.foodchem.2012.07.060.

Albuquerque, T.G., Oliveira, M.B.P.P., Sanches-Silva, A., Costa, H.S. (2016). Cholesterol Determination in Foods: Comparison Between High Performance and Ultra-High Performance Liquid Chromatography. Food Chem. 193, 18–25. doi:10.1016/j.foodchem.2014.09.109.

[AOAC] Association of Official Analytical Chemists. (2012a). AOAC Official Method 994.10 Cholesterol in Foods. Volum 1. Journal of AOAC International. Maryland (USA).

[AOAC] Association of Official Analytical Chemists. (2012b). AOAC Official Method 925.30 Solids (Total) in Eggs, Vacuum Method. Volum 1. Journal of AOAC International. Maryland (USA).

[AOAC] Association of Official Analytical Chemists. (2012c). Appendix K: Guidelines for Dietary Supplements and Botanical. Journal AOAC International. Maryland (USA).

Badawy, M.E.T.I. (2018). Development and Validation of HPLC Methods for Analysis of Chlorantraniliprole Insecticide in Technical and Commercial Formulations. Journal of Environmental Science and Health, Part B. 1-12. doi:10.1080/03601234.2018.143882 9.

Bavisetty, S.C.B., Narayan, B. (2015). An Improved RP-HPLC Method for Simultaneous Analyses of Squalene and Cholesterol Especially in Aquatic Foods. J. Food Sci. Technol. 52: 6083–6089. doi:10.1007/s13197-015-1793-4.

Bonfilio, R., Ricardo, C., Tarley, T., Ribeiro, G., Regina, H., Salgado, N., Benjamim, M., Araújo, D. (2009). Multivariate Optimization and Validation of An Analytical Methodology by RP-HPLC for The Determination of Losartan Potassium in Capsules. Talanta. 80: 236–241. doi:10.1016/j.talanta.2009.06. 060.

[BPS] Badan Pusat Statistik. (2017). Rata-rata Konsumsi Per Kapita Seminggu Beberapa Macam Bahan Makanan Penting, 2007-2016. [Internet]. Retrieved November 28, 2017 from: https://www.bps.go.id /linkTabelStatis/ view/id/950.

Cabo-calvet, E., Ortiz-bolsico, C., Baeza-baeza, J.J., García-alvarez-coque, M.C. (2014). Columns in Isocratic and Gradient Elution Using Mobile Phase Composition and Flow Rate as Factors. Chromatography. 194–210. doi:10.3390/chromatography1040194.

Chen, Y.Z., Kao, S.Y., Jian, H.C., Yu, Y.M., Li, J.Y., Wang, W.H., Tsai, C.W. (2015). Determination of Cholesterol and Four Phytosterols in Foods without Derivatization by Gas Chromatography-Tandem Mass Spectrometry. J. Food Drug Anal. 23: 636–644. doi:10.1016/j.jfda.2015.01.010.

Damodaran S, Parkin KL. (2017). Fennema's Food Chemistry. 5th edition. CRC Press. Boca Raton: USA.

Derewiaka D, Molinska E. (2015). Cholesterol transformations during heat treatment. Food Chemistry. 171: 233-240. doi: 10.1016/j.foodchem.2014.08.117.

Diana, R. (2013). Healthy Joints for Life: An Orthopedic Surgeon’s Plan to Reduce Pain and Inflamation, Avoid Surgery and Get Moving Again. USA: Harlequin.

Dinh TTN, Blanton JR, Brooks JC, Miller MF, Thompson LD. (2008). A Simplified Method for Cholesterol Determination in Meat and Meat Products. Journal of Food Composition and Analysis. 21: 306 – 314.

EURACHEM. (2014). Eurachem Guide: The Fitness for Purpose of Analytical Methods - A Laboratory Guide to Method Validation and Related Topics. 2nd Ed. Tersedia di : www.eurachem.org.

Exler, J., Philips, K.M., Patterson, K.Y., Holden, J.M. (2013). Cholesterol and Vitamin D of Eggs in The U.S. Retail Market. Journal of Food Composition and Analysis. 29: 110-116. doi: 10.1016/j.jfca.2012.11.001

Fletouris, D.J., Botsoglou, N.A., Psomas, I.E. (1998). Rapid Determination of Cholesterol in Milk and Milk Products by Direct Saponification and Capillary Gas Chromatography. J. Dairy Sci. 81: 2833–2840. doi:10.3168/jds.S0022-0302(98)75 842-4.

Gad, S.C. (2008). Preclinical Development Handbook: Adme Biopharmaceutical Properties. John willey and sons. Hoboken, New jersey.

Gulati S, Liu Y, Munkasci AB, Wilcox L, Sturley SL. (2010). Sterols and Sphingolipids: Dynamic Duo or Partners in Crime?. Prog Lipid Res. 49 (4): 353-365. doi:10.1016/j.plipres.2010.03.003.

Gorassini, A., Verardo, G., Fregolent, S., Bortolomeazzi, R. (2017). Rapid Determination of Cholesterol Oxidation Products in Milk Powder Based Products by Reversed Phase SPE and HPLC-APCI-MS/MS. Food Chem. doi:10.1016/j.foodchem.2017.03.080.

Haky JE. (2005). Resolution in HPLC: Selectivity, Efficiency, and Capacity. Di dalam: Cazes J, editor. Encyclopedia of Chromatography. 2nd Edition. Taylor & Francis Group. USA. Hlmn1434-1436.

Harris JR. (2010). Cholesterol Binding and Cholesterol Transport Proteins: Structure and Function in Health and Disease. Berlin (Germany): Springer Science & Bussiness Media.

[JECFA] Joint FAO/WHO Expert Committee on Food Additives. (2006). Combined Compendium of Food Additive Spesifications: Analytical Methods, Test Procedures and Laboratory Solutions Used by and Referenced in The Food Additive Specifications. Rome (Italy): Food and Agriculture Organization of The United Nations.

Kruve A, Rebane R, Kipper K, Oldekop M, Evard H, Herodes K, Ravio P, Leito I. 2015. Tutorial review on validation of liquid chromatography-mass spectrometry methods: part II. Analytica Chimica Acta. doi: http://dx.doi.org/10.1016/j.aca.2015.02.016.

Li, S., Tian, M., Row, K.H. (2010). Effect of Mobile Phase Additives on The Resolution of Four Bioactive Compounds by RP-HPLC. Int. J. Mol. Sci. 11: 2229-2240. doi:10.3390/ijms11052229.

Lin, Y., Wu, S., Wu, H. (2007). Highly Sensitive Analysis of Cholesterol and Sitosterol in Foods and Human Biosamples by Liquid Chromatography with Fluorescence Detection. J. Chromatogr. 1156: 280–287. doi:10.1016/j.chroma.2007.01.091.

Lioe, H.N., Setianingrum, T., Anggraeni, R. (2013). Validasi Metode Analisis Kolesterol dalam Telur dengan HPLC-ELSD. JIPI. 18 (3): 178-185.

Mariutti, L.R.B., Nogueira, C., Bragagnolo, N. (2008). Optimization and Validation of Analytical Conditions for Cholesterol and Cholesterol Oxides Extraction in Chicken Meat Using Response Surface Methodology. J. Agric Food Chem. 56: 2913-2918. doi: 10.1021/jf0735432.

Marquardt D, Kucerka N, Wassal SR, Harroun TA, Katsaras J. (2016). Cholesterol's Location in Lipid Bilayers. Chemistry and Physics of Lipids.

doi:10.1016/j.chemphyslip.2016.04.001.

Maslukhah, Y.L. (2019). Validasi Analisis Kolesterol Telur Ayam Broiler Menggunakan HPLC-MWD dan Aplikasinya pada Pangan dengan Matriks yang Sama. Unpublished master's thesis, Institut Pertanian Bogor, Bogor (ID).

Min, J., Lee, S., Khan, M.I., Yim, D.G., Seol, K., Lee, M., Jo, C. (2015). Monitoring The Formation of

Cholesterol Oxidation Products in Model Systems Using Response Surface Methodology. Lipids Health Dis. 14:77 (1–9). doi:10.1186/s12944-015-0074-6.

Naviglio, D., Gallo, M., Laura, L.G., Scala, C., Ferrara, C., Santini, A. (2012). Determination of Cholesterol in Italian Chicken Eggs. Food Chemistry. 132: 701-708. doi: 10.1016/j.foodchem.2011.11.002.

Niazi, S.K. (2016). Biosimilar and Interchangeable Biologics: Tactical Element. Boca Raton (Florida). CRC Press.

Ohvo-Rekila H, Ramstedt B, Leppimaki P, Slote JP. (2002). Cholesterol interactions with phospholipids inn membranes. Progress in Lipid Research. 41: 66-97.

Park, J., Jeong, I., Kwak, B., Ahn, J., Leem, D., Jeong, J., Kim, J. (2013). Application of Rapid Sample Preparation Method and Monitoring for Cholesterol Content in Chicken Egg and Egg powder. Korean J.Food Sci. An. 33: 672–677.

Poole, C.F., Poole, S.K. (2009). Foundations of Retention in Partition Chromatography. Journal of Chromatography A. 1216: 1530–1550. doi:10.1016/j.chroma.2008.10.092.

Rozner S, Garti N. (2006). The activity and absorption relationship of cholesterol and phytosterols. Colloids and Surfaces A: Physicochem. Eng. Aspects. 282-283: 435-456. doi: 10.1016/j.colsurfa.2005.12.032.

Sahu PK, Rao RN, Cecchi T, Swain S, Patro CS, Panda J. 2017. An overview of experimental designs in HPLC method development and validation. Journal of Pharmaceutical and Biomedical Analysis. doi: 10.1016/j.jpba.2017.05.006

Shabir GA. (2003). Validation of high-performance liquid chromatgraphy methods for pharmaceutical analysis: understanding the differences and similarities between validation requirements of the US Food and Drug Administration, the US Pharmacopeia and the International Conference on Harmonization. Journal of Chromatography A. 987: 57 - 66.

Song Y, Kenworthy AK, Sanders CR. (2014). Cholesterol as a co-solvent and a ligand for membrane proteins. The Protein Society. 23: 1-22. doi: 10.1002/pro.2385.

Souza, H.A.L., Mariutti, L.R.B., Bragagnolo, N. (2016). Microwave Assisted Direct Saponification for The Simultaneous Determination of Cholesterol and Cholesterol Oxides in Shrimp. J. Steroid Biochem. Mol. Biol. doi:10.1016/j.jsbmb. 2016.03.027.

Stroher, G.L., Rodrigues, A.C., Dias, L.F., Pedrão, M.R., De Paula, L.N., Visentainer, J.V., De Souza, N.E. (2012). Comparative Analysis and Validation Methodologies of GC and HPLC for Analysis of Cholesterol in Meat Products. AJAC. 3: 306–311.

Studzinska, S., Pietrzak, L., Buszewski, B. (2014). The Effects of Stationary Phases on Retention and Selectivity of Oligonucleotides in IP‑RP‑HPLC. Chromatographia. 77: 1589–1596. doi:10.1007/s10337-014-2766-x.

Smith, R.M. (2000). Handbook of Analytical Separations. Valko K, editor. Volume 1. Amsterdam: Elsevier Science BV.

Sunanti, S.T., Samin, Supriyanto, C. (2013). Penetapan Nilai Kandidat In-House Reference Material (RM) ZrO2. Jurnal Sains dan Teknologi Nuklir Indonesia. 14 (1): 23- 36.

Tiryaki O, Baysoyu D, Aydin G, Secer E. (2009). Setting system suitability parameters for performance optimization of GC-NPD detection fro pesticide residue analysis. GU Journal of Science. 22 (3): 149 - 155.

[USDA] United State Departement Agriculture. (2016). National Nutrient Database for Standars Reference Release 28: 01123, Egg, Whole, Raw, Fresh. [Internet]. [diunduh 2017 September 2]. Tersedia pada: https://ndb.nal.usda.gov/ndb/search/list/.

Wolfender JL. (2009). HPLC in natural product analysis: The detection issue. Planta Med. 75: 719-734. doi: 10.1055/s-0028-1088393.

DOI: http://dx.doi.org/10.31153/js.v22i2.795