Methods Validation of Pork Authentication in Processed Meat Products (Sausages) Through Densitometry Analysis


Sandra Hermanto(1*), Tarso Rudiana(2), Muhammad Ihda Hamlu Liwaissunati Zein(3), Asih Widi Wisudawati(4)

(1) Chemistry Study Program, Faculty of Science and Technology, Universitasi Islam Negeri (UIN) Syarif Hidayatullah Jakarta, Jl. Ir. H. Juanda No 95, Ciputat, Tangerang Selatan, Banten 15412
(2) Chemistry Study Program, Faculty of Science and Technology, Universitasi Islam Negeri (UIN) Syarif Hidayatullah Jakarta, Jl. Ir. H. Juanda No 95, Ciputat, Tangerang Selatan, Banten 15412 Department of Chemistry, Faculty of Science, Pharmacy and Health, Universitas Mathla’ul Anwar, Jl. Raya Labuan-Pandeglang KM. 23, Pandeglang, Banten 42273
(3) Chemistry Study Program, Faculty of Science and Technology, Universitasi Islam Negeri (UIN) Syarif Hidayatullah Jakarta, Jl. Ir. H. Juanda No 95, Ciputat, Tangerang Selatan, Banten 15412
(4) Institut für Didaktik der Chemie, Westfälische Wilhelms-Universität Münster, Raum Fl 219 Fliednerstr. 21, D-48149 Münster
(*) Corresponding Author

Abstract


Analysis of the halalness of food products based on animal origin needs to be carried out, especially for critical food products such as processed meat products (sausages). Some processed meat products are found that they are contaminated by pork in the processing. This research was conducted to determine the validity of the analysis method of pork contamination in one of the processed products (beef, pork, and goat sausages) based on the characteristics of the protein profile. Beef, pork, and goat sausages that have been purchased from supermarkets in Jakarta and South Tangerang are used as the sample.The praparation of sample was preceded by extraction of the sausage protein in PBS buffer pH 7.2. Then, the dissolved protein content was tested, and the protein extract was separated by SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis). The protein profile result from SDS-PAGE separation was followed by densitometric analysis (ImageJ) to obtain biomarker protein candidates. The validation of the analytical method includes the precision (repeatability), test for accuracy, linearity, and robustness of the method by varying the extraction pH. The results show that the protein extract has the differences in protein content and composition for each sample. The SDS-PAGE analysis results show that biomarker protein candidates appeared below 50 kDa which were thought to be the protein fraction of actin. The precision and accuracy test results obtained for each sample have met the required standards, namely with a KV value <5% and a percent recovery value> 95%. The results of the linearity test and the toughness of the method also show that the test method is quite effective in testing the halalness of animal food products, especially in sausage products which are contaminated by pork.


Keywords


densitometric; protein sausage; SDS-PAGE validation method

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References


Alonso Villela, S. M., Kraïem, H., Bouhaouala-Zahar, B., Bideaux, C., Aceves Lara, C. A., & Fillaudeau, L. (2020). A protocol for recombinant protein quantification by densitometry. MicrobiologyOpen, 9(6), 1175–1182. https://doi.org/10.1002/mbo3.1027

BPS. (2010). Penduduk Indonesia Menurut Wilayah dan Agama yang Dianut. Retrieved from https://sp2010.bps.go.id/index.php/site/tabel?tid=321

Erwanto, Y., Abidin, M. Z., Muslim, E. Y. P., Sugiyono, S., & Rohman, A. (2014). Identification of pork contamination in meatballs of Indonesia local market using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Asian-Australasian Journal of Animal Sciences, 27(10), 1487–1492. https://doi.org/10.5713/ajas.2014.14014

Fibriana, F., Widianti, T., Retnoningsih, A., & Susanti. (2012). Deteksi Daging Babi Pada Produk Bakso di Pusat Kota Salatiga Menggunakan Teknik Polymerase Chain Reaction. Biosaintifika: Journal of Biology & Biology Education, 4(2). https://doi.org/10.15294/biosaintifika.v4i2.3928

Harmita. (2004). Petunjuk Pelaksanaan Validasi Metode dan Cara Perhitungannya. Majalah Ilmu Kefarmasian, 1(3), 117–135.

Hermanto, S., & Meutia, C. D. K. (2009). Perbedaan Profil Protein Produk Olahan (Sosis) Daging Babi dan Sapi Hasil Analisa SDS-PAGE. Valensi, 1(4), 181–186.

Hermanto, S., Sholaikah, M. I., & Mulyani, S. S. (2016). Identification of Differentially Expressed Protein From. Molekul, 11(1), 71–79.

Laemmli, U. K. (1970). Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227, 680–685.

Listrat, A., Lebret, B., Louveau, I., Astruc, T., Bonnet, M., Lefaucheur, L., Picard, B., & Bugeon, J. (2016). How muscle structure and composition determine meat quality. The Scientific World Journal, 14(2), 1–14.

Maqsood, S., Manheem, K., Gani, A., & Abushelaibi, A. (2018). Degradation of myofibrillar, sarcoplasmic and connective tissue proteins by plant proteolytic enzymes and their impact on camel meat tenderness. Journal of Food Science and Technology, 55(9), 3427–3438. https://doi.org/10.1007/s13197-018-3251-6

Popovski, Z., Sciences, F., Blagica, T., Porchu, K., & Vukovic, V. (2002). New Approach in Detection of Porcine Stress Syndrome. Biotechnology in Animal Husbandry, 18(5-6), 73-80.

Rahmawati, Sismindari, Raharjo, T. J., Sudjadi, & Rohman, A. (2016). Analysis of pork contamination in Abon using mitochondrial D-Loop22 primers using real-time polymerase chain reaction method. International Food Research Journal, 23(1), 370–374.

Rohman, A., Himawati, A., Triyana, K., Sismindari, & Fatimah, S. (2017). Identification of pork in beef meatballs using Fourier transform infrared spectrophotometry and real-time polymerase chain reaction. International Journal of Food Properties, 20(3), 654–661. https://doi.org/10.1080/10942912.2016.1174940

Rokhim, S., Tyautari, I., Firmansyah, M. A., & Rachmawati, Y. (2021). End Point Polymerase Chain Reaction for Porcine Detection on Food Product of UIN Sunan Ampel Surabaya Canteen. Indonesian Journal of Halal Research, 3(1), 21–26. https://doi.org/10.15575/ijhar.v3i1.11149

Santoni, V., Molloy, M., & Rabilloud, T. (2000). Membrane proteins and proteomics: Un amour impossible? Electrophoresis, 21(6), 1054–1070. https://doi.org/10.1002/(SICI)1522-2683(20000401)21:6<1054::AID-ELPS1054>3.0.CO;2-8

Sarno, R., Triyana, K., Sabilla, S. I., Wijaya, D. R., Sunaryono, D., & Fatichah, C. (2020). Detecting Pork Adulteration in Beef for Halal Authentication using an Optimized Electronic Nose System. IEEE Access, 8, 25–30. https://doi.org/10.1109/ACCESS.2020.3043394

Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., & Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Anal Biochem, 150(1), 76-85.

Susanti, A. M., Darmawati, S., & Maharani, E. T. W. (2019). Five Types of Profile Meat Protein with Papaya Soaked Leaves Based SDS-PAGE. Gorontalo Journal of Public Health, 2(1), 132–138.

Susanto, E. (2005). Identifikasi Pencampuran Daging Dalam Baso. Retrieved from https://republika.co.id/berita/dunia-islam/info-halal/08/12/17/20904-identifikasi-pencampuran-daging-dalam-baso.

Wahniyathi, H., & Ali, H. M. (2005). Karakteristik protein daging dengan penambahan NaCl pada berbagai waktu aging post mortem dan hubungannya dengan mutu sensori sosis. Thesis. Fakultas Peternakan Universitas Hasanudin. Makassar.




DOI: https://doi.org/10.15575/ijhar.v4i1.11892

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Indonesian Journal of Halal Research Indexed By:

         


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Halal Center

UIN Sunan Gunung Djati

Gedung Solahuddin Sanusi (Laboratorium Terpadu)

Jl. A.H. Nasution No. 105, Cibiru, Bandung, West Java 40614 - Indonesia 

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Indonesian Journal of Halal Research by Halal Center UIN Sunan Gunung Djati Bandung is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Based on a work at https://journal.uinsgd.ac.id/index.php/ijhar.