Development of reference material for the detection of rat ingredient in meat by quantitative real-time PCR

A reference material (RM) for the determination of rat ingredient in meat was developed in the current study. The muscle tissues of thorax and leg of rat with fascia removed were used as the fundamental material, followed by freezing, slicing, drying, and grinding to obtain the meat powder. Cod was selected as the representative meat matrix and the developed RM was 1% w/w rat meat in cod meat. The reliability of the raw material to develop RM was firstly confirmed by quantitative real-time PCR. This RM was homogeneous and showed good stability during both long-term storage at 4 °C and short-term simulated transportation condition in the range of −20 °C - 45 °C. Characterization of this RM was conducted in eight collaborative laboratories. The characterized CT value of RM was 20.14 with an uncertainty of 1.26 (k = 2, where k is the coverage factor). This RM can be used for method validation and quality control to determine rat ingredient in meat products.