Using Box-Behnken design approach to investigate benzo[a]anthracene formation in smoked cattle meat samples and its' risk assessment.

In present study, a novel approach was selected for benzo[a]anthracene determination from smoked cattle meat samples using solid phase extraction under various experimental conditions. To optimize experimental variables, a three factor, three-level Box-Behnken experimental design combining with response surface methodology and quadratic programming were employed to prevent excessive amount of benzo[a]anthracene formation in smoked meat based on different experimental parameters. The influence of some important process parameters including cooking time (10-30 min), fat ratio (5-25%) and distance to the cooking source (5-25 cm), which significantly affected the formation efficiency of benzo[a]anthracene were optimized. The analysis of variance was conducted for specifying the interactions of independent variables. The independent and the dependent variables interactions were investigated. The quadratic regression model and the response surface contour plots were used to determine optimum values for the selected variables. The results of study revealed that optimum cooking time of smoked cattle meat was determined as 24.9 min, fat ratio 7.9% and the distance to the cooking source 21.8 cm. Under optimum conditions, minimum benzo[a]anthracene was formed in meat and its amount was determined using high performance liquid chromatography-mass spectrophotometer as qualitative and quantitative. The limit of detection and limit of quantitation values were calculated as 0.4 µg kg-1 and 1.1 µg kg-1, respectively.