A Capillary Electrophoresis Method for the Determination of Hydroxyproline as a Collagen Content Index in Meat Products

Abstract

The addition of collagen or its hydrolysates as a protein source or water binding agent is a common practice in the production of meat products. However, its level of addition is restricted by international regulations. Glycine, proline, and hydroxyproline are the most abundant amino acids in collagen, which is the main constituent of connective tissue. Thus, the objective of this study was to develop a method for the determination of hydroxyproline content in meat products by capillary electrophoresis (CE) as a collagen content index. Acid hydrolysis of samples and amino acid derivatization with fluorescamine prior to CE analysis were conducted. Separations were carried out using an uncoated capillary at 33 °C, using tetraborate buffer 0.05 M, pH 9.3 with 100 mM SDS. Hydroxyproline was detected at 214 nm. The assay was reproducible (RSD < 3% for normalized peak areas and migration times) and had good correlation (r = 0.975, p < 0.001) with the AOAC official method. From the samples tested (36), commercial ham showed the lowest average collagen content (<7.16 g per 100 g protein) by the CE method. On the other hand, frankfurter and Mexican “chorizo” sausages showed average collagen contents of 12.88 and 11.17 g per 100 g protein, respectively. The CE method developed could be used by regulatory agencies to ensure compliance with maximum levels of collagen addition in processed meats.