Introduction: The Cholesterol in our diet is a risk factor for the prevalence of cardiovascular diseases. One of the effective ways to reduce the incidence of such diseases is to remove cholesterol, including animal fats, from our food sources. In this research, attempts were made to investigate the effect of operational factors on reducing cholesterol in cream by beta-cyclodextrin using RSM and to examine its physicochemical properties. The variables in this study were beta-cyclodextrin concentration, mixing temperature and mixing time.
Materials and Methods: The 3.5% fat raw milk was pasteurized at 72°C for 16 s, and then it was cooled to 55°C. The milk cream was separated by a separator and adjusted to 36% fat. Beta-cyclodextrin was added in 0.5, 1, 1.5% concentration in water bath at three levels of 10, 30, 50 ° C. It was mixed at three time intervals of 5, 15, 25, min. Cream containing beta-cyclodextrin was centrifuged to eliminate the complex formed in different periods and the beta-cyclodextrin and cholesterol complex was isolated. Beta-cyclodextrin of 99.2% purity was purchased from Sigma-Aldrich and all chemical agents from Merk, Germany. To measure cholesterol content in the preparation phase, hexane and ethanolic solution of potassium hydroxide (Merk) and the standard solution of cholesterol (Merk) were used. To determine the level of cholesterol in the samples, Lee et al. (1999) method was used after a few modifications. Firstly, cholesterol was extracted from the specimens and the cholesterol levels in the samples were determined with a chromatography apparatus equipped with flame ionization. The quantitative measurement of cholesterol was conducted by comparing the peak areas with a response from the internal standard. The experiments were designed, using the response surface method and the Box-Behnken Design (BBD), to achieve a high-efficiency process in reducing the highest cholesterol level and determining optimum conditions. The viscosity was determined using a Brookfield viscometer, and the over run and foam stability of the cholesterol cream samples were determined using a Graduated cylinder.
Results and discussion: Due to the cavity in the central part of beta-cyclodextrin molecular arrangement, it can form a stable cholesterol-insoluble complex and help to isolate it from the product. The results showed that time, mixing temperature, and beta-cyclodextrin concentration all had a positive effect on reducing the level of cholesterol, and the effect of independent factors was almost the same, as the time factor had the highest effect, supported in Makoto et al. (1999). Concentration factor had the least effect on cholesterol reduction, contrary to studies by Aryafar et al. (2007) and Yen et al. (1995). Among the interactions of the investigated factors, the highest effect was due to the simultaneous effects of temperature and time (88.9%) and the least effect was due to the simultaneous effect of concentration and temperature (85.5%). Findings of this study showed that the use of beta-cyclodextrin resulted in a successful reduction of cholesterol from the product without affecting tissue properties, increasing viscosity and volume, or reducing stability of the foam. The absorption of cholesterol by beta-cyclosporine depends on its concentration. The mixing time and temperature also boosts the effect. After examining the effects of each independent variable, it can be concluded that all three investigated variables have an incremental effect on the level of cholesterol in the evaluated levels. Increasing mixing time has the highest effect on increasing cholesterol and increasing the concentration of beta-cyclodextrin has the lowest effect on cholesterol reduction. Investigating the interaction effects of variables also showed that the simultaneous effect of temperature and time with the proper concentration of beta-cyclodextrin could have a significant effect on cholesterol reduction. The response surface method and Box-Behnken Design as statistical optimization and modeling techniques enabled the researchers to predict the optimal conditions for maximal removal of cholesterol by beta-cyclodextrin at the decided levels. The optimal operating condition obtained by the model for 36% fat cream contains 42.1% beta-cyclodextrin concentration, 76.75 ° C temperature and 87.83 min of mixing time. The predicted cholesterol reduction in these conditions was 89.92%, which was very close to the experimental value obtained in predicted optimal conditions. It indicates the accuracy and predictive power of the intended model.