Food Engineering
Fateme Mousavi Baygi; Arash Koocheki; Behrooz Ghorani; Mohebbat Mohebbi
Abstract
Introduction Curcumin, as a natural polyphenolic nutraceutical has been shown many health-promoting effects, mainly associated with its chemical structure. In various studies, different properties of this compound, including anti-tumor and anti-cancer activity, reduction of blood and liver cholesterol ...
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Introduction Curcumin, as a natural polyphenolic nutraceutical has been shown many health-promoting effects, mainly associated with its chemical structure. In various studies, different properties of this compound, including anti-tumor and anti-cancer activity, reduction of blood and liver cholesterol levels, increase of immune function, prevention of cardiovascular diseases, prevention of damage to biological membranes against peroxidation and anti-inflammatory properties have been reported. Despite possessing a potential health benefits to humans, the susceptibility of this polyphenol towards environmental conditions and low chemical stability has restricted the direct usage of curcumin into aqueous-based food formulations. The encapsulation of curcumin in liposomes is a potentially effective way to protect them from degradation during passing the digestive system.Materials and MethodsCurcumin (powder, purity greater than 99%, 368.38 g/mol), lecithin, cholesterol (C3045-25G), pancreacin (extracted from porcine pancreas, P7545-25G), bile salts (B8756-10G) and calcium chloride (CaCl2) was obtained from Sigma Aldrich (USA). Consumable ethanol was purchased from Pars Ethanol Company (96%, Iran). Lipase enzyme (extracted from pig pancreas, L8070) and pepsin (activity 3500-3000 NFU/g, P8390) were obtained from Solarabio (China). Potassium chloride, dipotassium hydrogen phosphate (K2HPO4) and alpha-amylase enzyme with a purity of at least 99% were obtained from Merck, Germany, sodium chloride (NaCl), sodium bicarbonate (NaHCO3) and calcium chloride were obtained from Sigma. The effect of lecithin content (0.02- 0.08 g), lecithin cholesterol ratio (0.5- 4), curcumin level (1.5- 6mg) and ultrasound treatment time (1-5 minutes) on production of liposomes containing curcumin was evaluated. The particle size, particle size distribution, zeta potential and efficiency were determined by response surface methodology. Furthermore, physical nature, molecular structure, physical stability at 4ºC and 25ºC and release behavior of curcumin loaded-liposome in mouth, stomach and intestines were explored.Results and Discussion The results showed that all independent variables had a significant effect on liposome particle size and increasing the ratio of lecithin: cholesterol caused more uniform particle size. Lecithin was determined to be the only component affecting the zeta potential of liposome particles, and increasing the ultrasound time increased the efficiency of curcumin encapsulation in liposomes. The optimal point of liposome preparation conditions in the amount of 0.08 g lecithin, 4: 1 the ratio of lecithin: cholesterol, 4.16 mg curcumin and 5 minutes the ultrasound treatment was introduced by Design Expert software. In addition, curcumin was amorphous in optimal liposome spherical particles. Furthermore, the results of TEM showed that the liposomes are in the form of single-layer particles, spherical and without membrane rupture. This makes the bilayered nature of the vesicles clearly visible in this micrograph. The size of the particles obtained from this method was consistent with the data obtained from the dynamic light scattering method. From the results of infrared spectroscopy, it can be seen that curcumin is trapped in the liposome through hydrogen bonding in the double-layered vesicle of the liposome, the phenolic ring of curcumin with the phospholipid head group, as well as the hydrophobic interactions of the aromatic rings with the acyl phospholipid chains. Liposomes were more stable at refrigeration temperature. A very small amount of curcumin was released in the simulated oral phase, which is probably due to the short time and lack of specific enzymes to disrupt the phospholipid bilayers of the liposome. Although the pepsin enzyme is unable to penetrate the liposome membrane, acidic conditions change the angle of the head and tail groups of the lipids and lead to a change in the surface charge of the liposomes. The release of curcumin from liposome vesicles was greatly increased in the intestine. This sudden increase is due to the presence of bile salts as an emulsifying agent that can disrupt the phospholipid membrane and make the membrane more fluid. In addition, pancreatic lipase is adsorbed on the surface of lipids and then hydrolyzes the phospholipid into 2-acyl and 1-acyl lysophospholipids and free fatty acids. The release behavior of curcumin under gastrointestinal conditions was based on the Fick mechanism.
Sharareh Rezaeian; Hamid Reza Pourianfar; Shadi Bolourian; Hamed Saberian
Abstract
Introduction: At the present, culinary-medicinal mushroom Enoki, Flammulina velutipes, ranks fifth among cultivated mushrooms in the world. Enoki has a delightfully crunchy texture and desirable taste. In addition, research has proved that this mushroom possesses substantial nutritional and medicinal ...
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Introduction: At the present, culinary-medicinal mushroom Enoki, Flammulina velutipes, ranks fifth among cultivated mushrooms in the world. Enoki has a delightfully crunchy texture and desirable taste. In addition, research has proved that this mushroom possesses substantial nutritional and medicinal properties. However, commercialization of Enoki is not yet expanded worldwide and it is still largely behind the other major edible mushrooms in some countries in west Asia (such as Iran). Based on our recent successful study on cultivation of Enoki in locally available lignocellulosic substrates, this study was aimed to evaluate the possibility of production of a novel functional drink based on Enoki mushroom supplemented with curcumin. Materials and Methods: A pure culture of the cultivated strain of Enoki was freshly fructified in a locally available lignocellulosic substrate composed of 40 % wheat straw + 40 % sawdust + 18 % wheat bran + 1 % lime + 1 % gypsum. The fresh fruiting bodies were crushed to different forms of pulp and puree to be utilized in the basic formulation of drink. Then, different concentrations of mushroom particles (5, 7.5 and 10 %) and pectin gum (0.05, 0.075 and 0.10 %) were used. Following pasteurization at 90 ºC for one minute, the rheological and sensory characteristics of the drink were evaluated. The viscosity and the flow behavior of the samples were measured at 7°C. A rotational programmable viscometer (LVDV-II Pro, Brookfield Engineering Inc., USA) with an LV spindle was employed. About 25 ml of each drink was poured into the cylinder of the viscometer and shear rate was measured from 5 to 200 s−1 within 5 s intervals. Effect of different treatments (pectin gum and mushroom concentration) on color indices of the Enoki drink was studied using colorimeter (ColorFlex EZ, Hunter Lab, USA) and indices of, L* (brightness), a* (redness–greenness), b* (yellowness–blueness), were determined. Some sensory characteristics (taste, odor, color, mouth feel, consistency, particle concentration and overall acceptance) of the Enoki drinks were evaluated by 12 trained panelists (aged 25–50 years) using a 5 level hedonic test (1, 2, 3, 4 and 5) from left to right representing unacceptable, relatively acceptable, good, very good, and excellent. Three types of flavor (Lime, tropical fruits and apple) were employed and the effect of the flavors on odor, flavor, taste and overall acceptance of the pulp- based drink was investigated. Finally, three concentrations of curcumin (0.01, 0.03 and 0.05 %), as a natural colorant, were used in the pulp- based drink and the overall acceptance of the panelists was evaluated in a separate test. Factorial tests based on the completely randomized design was employed to evaluate the effect of different concentration of mushroom and pectin gum. Results were analyzed by analysis of variance (ANOVA) using SPSS 19 statistical software and the Duncan’s test with 95% confidence interval was used to compare the means of the tests. The results which were presented in this research have been obtained from the average values of 12 replicates. Results and Discussion: Rheological tests showed that the flow behavior of the drink was pseudoplastic (or shear thinning), which indicated that the viscosity was decreased when the shear rate increased. All the samples had a yield stress at 7°C. Therefore, all the drink samples had a jelly form before exerting any stress, while the mushroom particles were completely suspended in the drink. From the panelists’ point of view, there was not any difference between the color of the drink containing different gum and mushroom, but L*, a* and b* values were significantly different. By increasing of the gum and mushroom concentration, brightness index (L*) increased. Sensory tests indicated that the highest and the lowest scores of flavor were related to samples containing 5 and 10 % (w/w) mushroom, respectively (p<0.05). Finally, the drink containing 5% mushroom puree and 0.05 % pectin gum was selected. The results of sensory evaluation showed that the drink containing mushroom pulp was significantly better than one containing mushroom puree (p<0.05). The overall acceptance of the Enoki drink containing lime flavor was the highest and there were not any significant differences between those containing apple and tropical fruits flavor. Furthermore, sensory acceptance of the drink increased by adding 0.05 % curcumin into it. It can be concluded that the drink containing Enoki mushroom pulp may have a good potential to be accepted by consumers. The use of a culinary-medicinal mushroom such as Enoki along with a natural colorant (curcumin) may make the drink a functional food without having any side effect. Overall, the findings of this study showed that the industrial production of this novel functional drink, with acidic lime taste and curcumin colorant, is feasible from a technical point of view.
Maryam Ravaghi; Anna Maria Fadda
Abstract
Introduction: Curcumin is the main ingredient of turmeric spice, derived from rhizome of the Curcuma longa. It is a hydrophobic material with low bioavailability which is rapidly removed from the body due to its instability under physiological conditions and inadequate absorption. Curcumin is a polyphenol ...
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Introduction: Curcumin is the main ingredient of turmeric spice, derived from rhizome of the Curcuma longa. It is a hydrophobic material with low bioavailability which is rapidly removed from the body due to its instability under physiological conditions and inadequate absorption. Curcumin is a polyphenol and one of the most widely explored natural pharmacological agents in nanomedicine due to its high in vitro activity and low systemic bioavailability in vivo. In recent years, many efforts have been made to increase its therapeutic potential for disease prevention and health promotion, by using specific nano-delivery systems. Delivery and accumulation of curcumin in the intestine may represent an attractive strategy to improve its systemic absorption and bioavailability. The incorporation of curcumin in phospholipid vesicles (liposomes) may modulate its accumulation and be released, but do not improve its stability in the gastrointestinal environment due to the phospholipid degradation. An appropriate combination of phospholipid and polymer may provide a suitable strategy to protect the vesicles, resulting in an amelioration of curcumin intestinal deposition and bioavailability. In the present work, the potential use of Eudragit and hyaluronan to complex phospholipids was investigated, thus forming gastroresistant vesicles able to deliver the curcumin to the low intestine.
Materials and Methods: Soy phosphatidylcholine S75 (purity of 70%) and P90G (purity of 94%) were purchased from Lipoid GmbH (Germany). Sodium hyaluronate or hyaluronan with low molecular weight (200–400 kDa) was purchased from DSM Nutritional Products AG Branch Pentapharm (Switzerland). Eudragit S100 and L100 with molecular weight about 125 kDa were provided by Evonik Industries
AG (Germany). Curcumin and other chemicals were analytical grade and procured from Sigma-Aldrich (Italy). This paper was aimed to find a liposome type formulation to immobilize the phospholipid structures, and to protect liposomes and enhance curcumin stability from simulated gastrointestinal conditions. To this purpose, phospholipid (P90G and S75), sodium hyaluronate and Eudragit (S100 and L100) were combined to obtain immobilized liposomes and drug. Liposomes were formed by hydration, combined with sonication using a high intensity ultrasonic disintegrator (Soniprep 150 plus, UK). The average diameter, polydispersity index (PDI) and zeta potential were determined using a Zetasizer nano-ZS (Malvern Instruments, United Kingdom). Entrapment efficiency (EE%) was calculated as the percentage of the drug amount found after dialysis (Spectra/Por® membranes, 12–14 kDa MW cut off, 3 nm pore size, The Netherlands) versus that initially used. Curcumin content was estimated spectrophotometrically at λmax=424 nm by a Perkin Elmer UV/visible spectrophotometer (Lambda 25, USA) after disruption of liposomes. Vesicle formation and morphology were checked by cryogenic electron transmission microscopy, using a Tecnai F20 TEM (FEI Company, The Netherlands). The average diameter, PDI, zeta potential and EE% of the vesicles produced using Eudragit S100 and L100 were measured after 1 month incubation at 4°C to determine their stability. Vesicle behaviour was also measured in fluids at pH 2 and 7 and high ionic strength immediately after dilution, at 2h and 6 h. Data are expressed as mean±standard deviation (n=3 independent samples). Analysis of the data was made using SAS statistical software (version 9.0). Comparison among the means was carried out by Duncan’s multiple range test at p800 nm) and polydispersed (>0.8). In this work, the Eudragit was dispersed in ethanol with the curcumin, leading to a homogeneous, yellow transparent solution, and the curcumin and hyaluronan were dispersed in water. After freeze drying and rehydration with water, the vesicle size and polydispersity decreased, leading to the formation of more suitable systems. Liposomes of Eudragit were spherical or oval, multi-lamellar or large uni-lamellar vesicles with some smaller vesicles inside. These liposomes had high entrapment efficiency (>80%) immediately after production without significant decrease after storage. Liposomes of Eudragit S100 had smaller vesicles (287±31 nm) compared to those of Eudragit L100 (407±33 nm) after preparation Moreover, vesicles containing Eudragit S100 showed better stability during storage. Immobilization of liposomes in polymeric network of hyaluronan-Eudragit increased their stability against the mimicking conditions of gastrointestinal tract such as high ionic strength and pH variation. The present study indicated that the combination of P90G, Eudragit S100 and hyaluronan results in the production of innovative delivery systems with suitable dimensions and adequate stability, capable of protecting the curcumin from the pH variation during the stomach to colon transit, and favoring the local accumulation of the drug. Finally, a more detailed research according to ICH and WHO guidelines should be performed in a more advanced development step.
