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

S. Hermanto, T. Rudiana, Muhammad Ihda H L Zein, A. W. Wisudawati
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引用次数: 1

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.
方法采用密度分析法对肉制品(香肠)中猪肉的认证进行验证
需要对基于动物来源的食品的清真性进行分析,特别是对加工肉制品(香肠)等关键食品。一些加工肉制品被发现在加工过程中被猪肉污染。本研究旨在根据蛋白质谱特征确定加工产品(牛肉、猪肉和山羊香肠)中猪肉污染分析方法的有效性。从雅加达和南丹格朗超市购买的牛肉、猪肉和山羊香肠被用作样本。样品制备前,在pH为7.2的PBS缓冲液中提取香肠蛋白。然后检测溶解蛋白的含量,并用SDS-PAGE(十二烷基硫酸钠聚丙烯酰胺凝胶电泳)对蛋白提取物进行分离。SDS-PAGE分离得到的蛋白谱随后进行密度分析(ImageJ)以获得候选生物标记蛋白。分析方法的验证包括精密度(可重复性)、准确度测试、线性测试和通过改变提取ph的方法的鲁棒性。结果表明,每个样品的蛋白质提取液在蛋白质含量和组成上存在差异。SDS-PAGE分析结果显示,生物标志物候选蛋白出现在50 kDa以下,被认为是肌动蛋白的蛋白部分。每个样品的精密度和准确度测试结果均达到要求标准,即KV值达到95%。线性测试结果和方法的韧性测试结果也表明,该测试方法对动物性食品的清真度测试是相当有效的,特别是对猪肉污染的香肠产品。
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