Effects of different crushing and refining treatments on release of carotenoids in carrot juice

Abstract: Carotenoids have received special attention owing to their health-related characteristics, including pro-vitamin A activity, antioxidant properties and prevention organisms against certain types of cancer. In carrot, the existence of chromoplast substructure and cell wall limit the release of carotenoids, thus adopting appropriate mechanical processing to destroy the cell wall and decrease the particle size is an effective way to improve carotenoid bioaccessibility. As the first step of digestion, the release of carotenoids from raw materials plays a crucial role in determining carotenoid bioaccessibility. The objective of this study was to investigate the effects of different crushing and refining treatments on release pathway of carotenoids in carrot juice. Pulping, colloid mill, and high pressure homogenization with different homogenization pressures, homogenization cycles and inlet temperatures were applied as crushing and refining treatments. After different crushing and refining treatments, carotenoid content in different organizational states (large cell clusters, small cell clusters and chromoplast fraction), free carotenoid content, micrograph of carotenoid distribution, release rate of carotenoids, as well as contents of oil-extracted carotenoids, β-carotene and α-carotene were investigated, respectively. Total carotenoid content was determined spectrophotometrically. β-carotene and α-carotene content were measured by HPLC. Micrograph of carotenoid distribution was observed by confocal laser scanning microscopy. The results demonstrated that after crushing treatment, carotenoid content of above-mentioned four fractions was in a decreasing order: free carotenoid, large cell clusters, small cell clusters and chromoplast fraction. After homogenization, there was almost no difference in free carotenoid content, however, there were about fourfold increase in carotenoid content for chromoplast fraction, twofold increase in carotenoid content for small cell clusters, and twofold decrease for large cell clusters. The proportion of carotenoid content decreased from 36.18% to 15.28% in large cell clusters, increased from 8.36% to 30.56% in chromoplast fraction and increased from 11.11% to 15.73% in small cell clusters. Therefore, an enhancement of carotenoid content could be observed with the decreasing level of bio- encapsulation. After refining treatments, the visible carotenoid particles under laser scanning confocal microscopy decreased in size. As the homogenization pressure increased up to 100 MPa and 150 MPa, confocal micrographs showed that visible carotenoid particles became smaller than the ones treated by other homogenization pressures. When the pressure reached 180 MPa, carotenoid tended to aggregate. The visible particle became smaller with the increase of homogenization cycle. Increasing inlet temperature contributed to aggregation of carotenoid particles. The value of carotenoid release rate was 50.91% for joint processing of colloid mill and high pressure homogenization. However, different levels of homogenization pressure had no significant (P>0.05) effect on carotenoid release rate. Higher carotenoid release rate could be observed with an increment of homogenization cycle and inlet temperature. Therefore, suitable refining treatment can be found and applied to damage the cell wall and effectively promote the release of carotenoids in carrot juice. As lipophilic pigment, carotenoid must first be released from the food matrix, solubilized in the lipid phase followed by transferring into mixed micelles in the small intestine before the uptake by intestinal epithelium. Therefore, oil-extracted carotenoid content reflects the available dose that may be absorbed by human body. The highest oil-extracted carotenoid content in carrot juice was 34.79 μg/g treated by pulping. When the pressure increased from 10 MPa to 180 MPa, the content of oil-extracted carotenoids decreased at first and then increased. The value of oil-extracted carotenoid content reached 31.93 μg/g for 100 MPa and 31.5 μg/g for 150 MPa, respectively. Contrarily, the value was 27.07 μg/g for homogenization at 180 MPa. The highest contents of oil-extracted β-carotene and α-carotene were 32.61 μg/g and 6.64 μg/g, respectively. Under the different conditions of homogenization, the content of β-carotene and α-carotene changed differently due to structural differentiation. This study has significance on subsequent analysis about how to improve carotenoid bioaccessibility in carrot juice. After refining treatment, polysaccharides such as pectin may be released from the cell wall, wrap around the carotenoid and affect the release of the carotenoid. Therefore, further research is needed to study the interaction between carotenoid and endogenous pectin in carrot juice after high homogenization.