Micelle separation conditions based on particle size strongly affect carotenoid bioaccessibility assessment from juices after in vitro digestion

For assessing the carotenoid bioaccessibility during in vitro digestion, obtaining the micelle fraction that can diffuse through the mucin layer according to the particle size is an important step. However, the accuracy of the various approaches to obtaining the right fraction of micelles described in literature has not been investigated. In many studies the reported bioaccessible fraction might therefore be over- or underestimating the real bioaccessibility. The present study aimed to fill this gap and give approaches to get accurate data on the bioaccessible fraction from samples with different properties. Results illustrated that optimizing the centrifugal speed and duration of the digesta are essential in obtaining the micelle fraction. Different digesta have different optimum centrifugal parameters to obtain the bioaccessible fraction. 6,000 rpm and 40 min was selected as the optimum centrifugal parameter for combined juice (CJ). 6,000 rpm and 60 min was the optimum centrifugal parameter for CJ with oil, and CJ with emulsion using non-ionic emulsifier. 8,000 rpm and 20 min was the optimum centrifugal parameter for CJ with emulsion using ionic emulsifier. Polygalacturonanase (PG)-digested systems required higher centrifugal speed (10,000 rpm) compared with CJ-based systems (6,000 rpm or 8,000 rpm). A prediction model to determine the optimal centrifugation speed/time from the properties of the intestinal digesta was developed. Sample preparation conditions strongly affect carotenoid bioaccessibility assessment from juices during in vitro digestion. Based on these results, it is highly recommended to perform an optimized preparation procedure for bioaccessible fraction prior to carotenoid bioaccessibility analysis.