Digestion of lipid excipients and lipid-based nanocarriers by pancreatic lipase and pancreatin.

The digestion behaviour of lipid-based nanocarriers (LNC) has a great impact on their oral drug delivery properties. In this study, various excipients including surfactants, glycerides and waxes, as well as various drug-delivery systems, namely self-emulsifying drug delivery systems (SEDDS), solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) were examined via the pH-stat lipolysis model. Lipolysis experiments with lipase and pancreatin revealed the highest release of fatty acids for medium chain glycerides, followed by long chain glycerides and surfactants. Waxes appeared to be poor substrates with a maximum digestion of up to 10% within 60 min. Within the group of surfactants, the enzymatic cleavage decreased in the following order: glycerol monostearate > polyoxyethylene (20) sorbitan monostearate > PEG-35 castor oil > sorbitan monostearate. After digestion experiments of the excipients, SEDDS, SLN and NLC with sizes between 30 and 300 nm were prepared. The size of almost all formulations was increasing during lipolysis and levelled off after approximately 15 min except for the SLN and NLC consisting of cetyl palmitate. SEDDS exceeded 6000 nm after some minutes and were almost completely hydrolysed by pancreatin. No significant difference was observed between comparable SLN and NLC but surfactant choice and selection of the lipid component had an impact on digestion. SLN and NLC with cetyl palmitate were only digested by 5% whereas particles with glyceryl distearate were decomposed by 40-80% within 60 min. Additionally, the digestion of the same SLN or NLC, only differing in the surfactant, was higher for SLN/NLC containing polyoxyethylene (20) sorbitan monostearate than PEG-35 castor oil. This observation might be explained by the higher PEG content of PEG-35 castor oil causing a more pronounced steric hindrance for the access of lipase. Generally, digestion experiments performed with pancreatin resulted in a higher digestion compared to lipase. According to these results, the digestion behaviour of LNC depends on both, the type of nanocarrier and on the excipients used for them.

The Procyanidin C1-Dependent Inhibition of the Hydrolysis of Potato Starch and Corn Starch Induced by Pancreatin.

Procyanidins are contained in various foods, and their effects on starch hydrolysis have been reported. In Japan, black soybeans, which contain a trimeric procyanidin, procyanidin C1 (proC1), are cooked with rice and used to prepare dumplings. In this study, the effects of proC1 on the pancreatin-induced formation of reducing sugars and starch hydrolysis were studied using potato starch and corn starch. ProC1 inhibited both reactions; the inhibition was greater in potato starch than corn starch when added to heated potato starch and corn starch. When heated with proC1, its inhibitory effects decreased, especially in potato starch, suggesting the important role of proC1 itself for the inhibition of potato starch hydrolysis. ProC1 also inhibited the hydrolysis when added to heated, longer amylose (average molecular weight: 31,200), and the inhibition decreased when heated with the amylose. On the other hand, proC1 could not inhibit the hydrolysis when added to heated, shorter amylose (average molecular weight: 4500), but could when heated with the amylose, suggesting the important role of the degradation products of proC1 for the inhibition. We discuss the mechanism of the proC1-dependent inhibition of amylose hydrolysis, taking the molecular weight into account.

In vitro Comparison of Pancreatic Enzyme Preparations Available in the Indian Market.

PURPOSE: Pancreatic enzyme replacement therapy (PERT) involves exogenous enzyme supplementation and is used in the treatment of pancreatic exocrine insufficiency. Clinical efficacy of PERT preparations is a function of physical properties and release kinetics that vary between commercially available products. In this study, we evaluated the physical properties, in vitro dissolution, and release kinetics of commercially available pancreatic enzyme preparations available in the Indian market. METHODS: Physical properties such as particle size distribution and water content of the capsules were measured by dynamic light scattering and Karl-Fischer titration method, respectively. An analytical procedure based on the European pharmacopoeia (EP) method was used to determine lipase activity, and a modified United States pharmacopoeia (USP)-based method was used for dissolution studies. Enzyme release was ascertained under gastroduodenal conditions in buffered media. RESULTS: Considerable variations in physical properties such as particle size and water content were observed between pancreatic enzyme preparations. Some preparations failed to meet the labeled lipase content as per USP standards (>90% label claim) and showed inconsistent release behavior (>5% relative standard deviation). CONCLUSION: Differences exist between pancreatic enzyme preparations in terms of physical properties, dissolution, and release behavior that can affect their clinical efficacy. The present study suggests, therefore, that these preparations should not be used interchangeably.

Enzymatic Preparation of Bioactive Peptides Exhibiting ACE Inhibitory Activity from Soybean and Velvet Bean: A Systematic Review.

The Angiotensin-I-converting enzyme (ACE) is a peptidase with a significant role in the regulation of blood pressure. Within this work, a systematic review on the enzymatic preparation of Angiotensin-I-Converting Enzyme inhibitory (ACEi) peptides is presented. The systematic review is conducted by following PRISMA guidelines. Soybeans and velvet beans are known to have high protein contents that make them suitable as sources of parent proteins for the production of ACEi peptides. Endopeptidase is commonly used in the preparation of soybean-based ACEi peptides, whereas for velvet bean, a combination of both endo- and exopeptidase is frequently used. Soybean glycinin is the preferred substrate for the preparation of ACEi peptides. It contains proline as one of its major amino acids, which exhibits a potent significance in inhibiting ACE. The best enzymatic treatments for producing ACEi peptides from soybean are as follows: proteolytic activity by Protease P (Amano-P from Aspergillus sp.), a temperature of 37 °C, a reaction time of 18 h, pH 8.2, and an E/S ratio of 2%. On the other hand, the best enzymatic conditions for producing peptide hydrolysates with high ACEi activity are through sequential hydrolytic activity by the combination of pepsin-pancreatic, an E/S ratio for each enzyme is 10%, the temperature and reaction time for each proteolysis are 37 °C and 0.74 h, respectively, pH for pepsin is 2.0, whereas for pancreatin it is 7.0. As an underutilized pulse, the studies on the enzymatic hydrolysis of velvet bean proteins in producing ACEi peptides are limited. Conclusively, the activity of soybean-based ACEi peptides is found to depend on their molecular sizes, the amino acid residues, and positions. Hydrophobic amino acids with nonpolar side chains, positively charged, branched, and cyclic or aromatic residues are generally preferred for ACEi peptides.

Differences in In Vitro Properties of Pancreatin Preparations for Pancreatic Exocrine Insufficiency as Marketed in Russia and CIS.

BACKGROUND: Pancreatic enzyme-replacement therapy (PERT), provided as pancreatin to patients with pancreatic exocrine insufficiency (PEI), is considered an essential substitute for the pivotal physiological function the pancreas fulfills in digestion. PEI involves a reduction in the synthesis and secretion of pancreatic enzymes (lipase, protease, amylase), which leads to an inadequate enzymatic response to a meal and consequently to maldigestion and malabsorption of nutrients. The efficacy of PERT is strongly dependent on enzyme activity, dissolution, and pancreatin particle size. OBJECTIVE: The physiological properties of eight pancreatin preparations (nine batches; five different brands) available in Russia and CIS (Commonwealth of Independent States: Armenia, Azerbaijan, Belarus, Kazakhstan, Kyrgyzstan, Moldova, Russia, Tajikistan, Uzbekistan) were investigated. METHODS: The lipase activity, dissolution, and particle size distribution of samples from multiple batches of pancreatin of different strengths were measured. RESULTS: Regarding lipase activities, all pancreatin preparations except Micrazim® matched the labeled content. Considerable differences were observed in particle size and dissolution. CONCLUSION: Pancreatin preparations available in Russia and CIS demonstrate product-to-product and batch-to-batch variability regarding the measured properties of lipase activity, dissolution, and particle size. This may impact the efficacy of PERT and therefore clinical outcomes.

Influence of Sonication and Taraxacum Officinale Addition on the Antioxidant and Anti-ACE Activity of Protein Extracts from Sous Vide Beef Marinated with Sour Milk and after In Vitro Digestion.

The present study assessed the effect of pretreating beef as a raw material for sous vide steak preparation. The pretreatment involved maceration of a batch of meat in sour milk with the simultaneous use of ultrasound (250 or 500 W) as well as the addition of Taraxacum officinale. The biological activity profile of the peptides was assessed in terms of their antioxidant activity and inhibiting activity against angiotensin-converting enzyme (ACE). Changes in the biological activity of peptides under the influence of hydrolysis by gastrointestinal enzymes, i.e., pepsin and pancreatin, were also considered. There was no significant effect of T. officinale addition and sonication of beef batches on the protein content (except for lot S6, after sonication at 500 W as acoustic power and with the addition of dandelion). It was observed that the interaction of maceration in sour milk with simultaneous ultrasound treatment as the initial production step of sous vide beef steak generates the formation of peptides with antioxidant properties. Moreover, peptide formation can be further enhanced by adding dandelion (based on the results of antiradical and chelating activity tests). In addition, the progression of hydrolysis under the influence of gastrointestinal enzymes promotes the release of peptides with antioxidant and anti-ACE activity.

The potential application of the protein hydrolysates of three medicinal plants: cytotoxicity and functional properties.

Functional evaluation of encrypted bioactive peptides in protein structure helps to better understand those for using in pharmacy and food sciences. For this purpose, the total protein was extracted from Matricaria chamomilla, Ziziphora clinopodioides, and Cressa cretica, and partially purified with ammonium sulfate. Protein hydrolysates were obtained from pancreatin hydrolysis for 240 min and the enzyme hydrolysis was confirmed using the determination of hydrolysis degree and Fourier transform infrared (FT-IR) followed by the physicochemical and sensory properties were investigated. The results showed that all hydrolysates had both cytotoxic and antioxidant activities. Specifically, C. cretica hydrolysates represented cytotoxic activity against the MCF-7 cell line with the IC50 of 135.21 µg/mL, while showed no significant growth inhibition effect on the HEK293 cell line. Besides, M. chamomilla hydrolysates showed the lowest bitterness value (1.125 ± 0.52). From the perspective of color investigation, M. chamomilla hydrolysates indicated the highest L* and the lowest a* factors. The highest turbidity and surface tension, and 10-fold more cancer cell killing effect under gastrointestinal digestion conditions were observed for M. chamomilla hydrolysates. Therefore, bioactive peptides might be formulated in designing of novel anticancer drugs or could be used in promising protocols for the production of food products with beneficial health effects.

The Effect of Enzymes and Sodium Lauryl Sulfate on the Surface Tension of Dissolution Media: Toward Understanding the Solubility and Dissolution of Carvedilol.

The objective of this work was to study the effect of the physiologically relevant enzymes pepsin, pancreatin, and the synthetic surfactant sodium lauryl sulfate (SLS) on the surface tension of the dissolution media and the solubility and dissolution of the weakly basic drug carvedilol. Compendial dissolution media and buffer solutions that simulate the gastrointestinal fluid, prepared with and without the addition of SLS, were used in this study. The surface tension of the dissolution media; critical micelle concentration (CMC) of SLS in buffer solutions; and size, polydispersity index, and zeta potential of SLS micelles loading carvedilol were determined. The solubility and dissolution of carvedilol were investigated and compared with those of the corresponding media prepared without the addition of pepsin, pancreatin, and SLS. Results showed that the addition of pepsin, pancreatin, and SLS lowered the surface tension of the dissolution media to 54.8, 55.7, and ~ 30 mN/m, respectively. The solubility of carvedilol was significantly enhanced with pepsin and SLS; however, no significant difference was found with pancreatin. The dissolution rate of carvedilol was fast in simulated gastric fluid with and without pepsin. The dissolution was further enhanced in media with pancreatin and SLS. The dissolution data were corroborated with the molar micellar solubilization (X) of SLS, ranging between 0.02 and 3.09. Understanding the effect of pepsin, pancreatin, and SLS on the surface tension of the dissolution media and the solubility and dissolution of poorly soluble drugs can improve our knowledge of the performance of these drugs in vivo.

Supersaturated-Silica Lipid Hybrids Improve in Vitro Solubilization of Abiraterone Acetate.

PURPOSE: Abiraterone acetate (AbA) is a poorly water-soluble drug with an oral bioavailability of <10% and a significant pharmaceutical food effect. We aimed to develop a more efficient oral solid-state lipid-based formulation for AbA using a supersaturated silica-lipid hybrid (super-SLH) approach to achieve high drug loading, improve in vitro solubilization and mitigate the food effect, while gaining a mechanistic insight into how super-SLH are digested and release drug. METHODS: The influence of super-SLH saturation level and lipid type on the physicochemical properties and in vitro solubilization during lipolysis of the formulations was investigated and compared to the commercial product, Zytiga. RESULTS: Super-SLH achieved significantly greater levels of AbA solubilization compared to Zytiga. Solubilization was influenced by the AbA saturation level, which determined the solid state of AbA and the relative amount of lipid, and the lipid utilized, which determined its degree of digestion and the affinity of the lipid and digestion products to the silica. A fine balance existed between achieving high drug loads using supersaturation and improving performance using the lipid-based formulation approach. The non-supersaturated SLH prepared with Capmul PG8 mitigated the 3-fold in vitro food effect. CONCLUSION: SLH and super-SLH improve in vitro solubilization of AbA, remove the food effect and demonstrate potential to improve oral bioavailability in vivo. Graphical Abstract Abiraterone acetate was formulated as silica-lipid hybrids and demonstrated enhanced in vitro solubilization in comparison to pure abiraterone acetate and commercial product, Zytiga.

Antibacterial, anti-biofilm activity and mechanism of action of pancreatin doped zinc oxide nanoparticles against methicillin resistant Staphylococcus aureus.

Staphylococcus aureus are known to cause diseases from normal skin wound to life intimidating infections. Among the drug resistant strain, management of methicillin resistant Staphylococcus aureus (MRSA) is very difficult by using conventional antibiotic treatment. Both Zinc oxide nanoparticles (ZnONPs) and pancreatin (PK) are known to have antibacterial activity. Our main objective is to dope PK on ZnONPs to reduced zinc-oxide toxicity but increased anti-bacterial and anti-biofilms activity. In present study, we showed that, functions of zinc oxide nanoparticles with pancreatin enzyme (ZnONPs-PK) have anti-bacterial, anti-biofilms, anti-motility and anti-virulence properties against MRSA. Moreover, ZnONPs-PK were more potent to eradicate MRSA than only ZnONPs and PK. Application of the produced nano-composites as treatment on infected swine dermis predominantly reflects the potential treatment property of it. The vancomycin sensitivity of MRSA was significantly increased on application with ZnONPs-PK. Further study revealed cell membrane was the target of the ZnONPs-PK and that leads to oxidative damage of the cells. The produced nanoparticles were found completely non-toxic to human's keratinocytes and lung epithelial cell lines at its bactericidal concentration. Overall, this study emphasizes the potential mechanisms underlying the selective bactericidal properties of ZnONPs-PK against MRSA. This novel nanoparticle strategy may provide the ideal solution for comprehensive management of MRSA and its associated diseases with minimising the use of antibiotics.

Characterization of all the lipolytic activities in pancreatin and comparison with porcine and human pancreatic juices.

Porcine pancreatic extracts (PPE), also named pancreatin, are commonly used as a global source of pancreatic enzymes for enzyme replacement therapy in patients with exocrine pancreatic insufficiency. They are considered as a good substitute of human pancreatic enzymes and they have become a material of choice for in vitro models of digestion. Nevertheless, while the global PPE contents in lipase, protease and amylase activities are well characterized, little is known about individual enzymes. Here we characterized the lipase, phospholipase, cholesterol esterase and galactolipase activities of PPE and compared them with those of porcine (PPJ) and human (HPJ) pancreatic juices. The phospholipase to lipase activity ratio was similar in PPJ and HPJ, but was 4-fold lower in PPE. The galactolipase and cholesterol esterase activities were found at lower levels in PPJ compared to HPJ, and they were further reduced in PPE. The enzymes known to display these activities in HPJ, pancreatic lipase-related protein 2 (PLRP2) and carboxylester hydrolase/bile salt-stimulated lipase (CEH/BSSL), were identified in PPJ using gel filtration experiments, SDS-PAGE and LC-MS/MS analysis. The galactolipase and cholesterol esterase activities of PPE indicated that PLRP2 and CEH/BSSL are still present at low levels in this enzyme preparation, but they were not detected by mass spectrometry. Besides differences between porcine and human enzymes, the lower levels of phospholipase, galactolipase and cholesterol esterase activities in PPE are probably due to some proteolysis occurring during the production process. In conclusion, PPE do not provide a full substitution of the lipolytic enzymes present in HPJ.

Formation of fibrils derived from whey protein isolate: structural characteristics and protease resistance.

The formation of lactoglobulin (ß-lg) fibrils from its native globular form has been investigated hitherto with pure lactoglobulin conditions. For commercially feasible production of ß-lg fibrils for food and therapeutic applications, the effect of impurities (albeit at very small fractions) found in industrial grade lactoglobulin proteins will need to be considered. In this work, food grade whey protein isolate (WPI), with its associated trace globulin and other impurities, was used to study the ß-lg fibril formation process. SDS-PAGE analysis supported by Thioflavin T measurement showed that the WPI fibril formation accompanied by hydrolysis limited the formation reaction rate; further analysis indicated that main impurities especially for α-lactalbumin may affect the intermolecular contacts among ß-lg. The increase of ß-sheet and α-helix conformation analyzed by circular dichroism revealed that the ß strand axis with an α-helix stabilized the linear structure of fibrils. The WPI fibrils exhibited a significant variation in lengths and differences in morphology when compared to some other protein fibrils reported in the literature. Two-stage pepsin followed by pancreatin digestion analysis revealed that the WPI fibrils produced will have the potential as a medium for active compound delivery to the intestinal environment.

Starch digestion in intact pulse cells depends on the processing induced permeability of cell walls.

Enzyme digestion of starch granules encapsulated within cell walls in pulse depends on both the intactness of cellular structure as well as the retention of the ordered structure of starch during processing. However, the role of cell wall permeability in starch digestion, as affected by processing conditions, has not been fully elucidated. In this study, Kubali bean cells were isolated under different processing conditions (i.e., high pressure-heating, hydrothermal processing, and acid-alkali treatments) individually and in combinations to elucidate the structure and in vitro digestion of entrapped starches in the cells. The morphological features and crystalline structure of entrapped starches suggest that intact cell walls hinder the starch gelatinization, which in turn lowers the enzyme susceptivity. Alteration of cell wall permeability induced by different processing conditions is further evaluated through the diffusion of fluorescence-tagged dextran probes. Among all the treatments, cells isolated with the pressure-cooking method exhibited higher cell wall permeability. The study suggests that the in vitro starch digestion of plant foods can be optimized through the selection of processing method that has the least impact on cell wall permeability.

Drug-carrier binding and enzymatic carrier digestion in amorphous solid dispersions containing proteins as carrier.

In order to further explain the ability of gelatin 50PS and bovine serum albumin (BSA) to generate supersaturation of a series of poorly soluble drugs (carbamazepine, cinnarizine, diazepam, itraconazole, nifedipine, indomethacin, darunavir (ethanolate), ritonavir, fenofibrate, griseofulvin, ketoconazole, naproxen, phenylbutazone and phenytoin), drug-polymer binding was investigated using solution NMR and equilibrium dialysis experiments. Binding characteristics of the biopolymers were compared to those of PVP, PVPVA and HPMC. Since both biopolymers are prone to enzymatic digestion, we evaluated the influence of proteolytic enzymes like pepsin and pancreatin on the dissolution properties of poorly soluble compounds when formulated as amorphous solid dispersions with gelatin 50PS and BSA. Evidence is being presented that supports the importance of drug-polymer binding in inducing and stabilizing supersaturation of poorly soluble drugs and enhancing dissolution from ASDs. In fact, BSA displayed drug binding with nearly all tested model drugs while in case of gelatin 50PS binding was observed for 5 out of 12 drugs. Addition of pepsin or pancreatin during dissolution of the biopolymer-containing ASDs leads to a drop in the concentration of the drug pointing to enzymatic digestion of the gelatin and BSA. However, after digestion, these formulations still outperformed their crystalline counterparts.

Enzymatic hydrolysis as a sample pre-treatment for titanium dioxide nanoparticles assessment in surimi (crab sticks) by single particle ICP-MS.

A reliable sample pre-treatment based on enzymatic hydrolysis has been fully optimized and validated for TiO2 NPs isolation and determination/characterization in surimi (crab sticks). Efficient extractions have been found when using a pancreatin/lipase mixture at pH 7.4 and 37 °C for 12 h under continuous stirring. The proposed sample pre-treatment procedure has been found not to change TiO2 NPs size distribution, therefore guaranteeing TiO2 NPs integrity. TiO2 NPs determination (TiO2 NPs concentration) and TiO2 NPs characterization (size distribution) were assessed by single-particle inductively coupled plasma mass spectrometry (sp-ICP-MS) working with dwell times in the microsecond range (high frequency of data acquisition). Method validation was performed for TiO2 NPs concentrations and TiO2 NPs sizes. Good repeatability (25% and 8% for TiO2 NPs concentration and TiO2 NPs most frequent size), and sensitivity (limit of detection of 5.28 × 105 NPs g-1for TiO2 NPs concentrations, and 31.3-37.1 nm for TiO2 NPs size) were obtained. Accuracy, calculated through analytical recovery was adequate. Recoveries for TiO2 NPs standards of 50 and 100 nm were 108 ±â€¯5 and 105 ±â€¯4%, respectively. The proposed methodology was applied to several surimi samples for assessing TiO2 NPs concentrations and size distribution. Some surimi samples were found to contain TiO2 NPs (concentrations from 1.40 × 107 to 1.19 × 109 NPs g-1). TiO2 NPs size distributions were very different among the samples, and some of them showed wide size ranges (the most frequent size varied from 53.8 to 62.1 nm; whereas, the mean size values were within the 73.4-217.5 nm range).

Release of Indospicine from Contaminated Camel Meat following Cooking and Simulated Gastrointestinal Digestion: Implications for Human Consumption.

Indospicine, a hepatotoxic arginine analog, occurs in leguminous plants of the Indigofera genus and accumulates in the tissues of grazing animals that consume these plants. Furthermore, indospicine has caused toxicity in dogs following consumption of indospicine-contaminated meat; however, the potential impact on human health is unknown. The present study was designed to determine the effect of simulated human gastrointestinal digestion on the release and degradation of indospicine from contaminated camel meat following microwave cooking. Results showed no significant (p > 0.05) indospicine degradation during cooking or in vitro digestion. However, approximately 70% indospicine was released from the meat matrix into the liquid digesta during the gastric phase (in the presence of pepsin) and increased to >90% in the intestinal phase (with pancreatic enzymes). Following human consumption of contaminated meat, this soluble and more bioaccessible fraction of intact indospicine could be readily available for absorption by the small intestine, potentially circulating indospicine throughout the human body to tissues where it could accumulate and cause detrimental toxic effects.

In vitro digestibility of normal and waxy corn starch is modified by the addition of Tween 80.

Aqueous dispersions of normal and waxy corn starch (3% w/w) were mixed with Tween 80 (0, 7.5, 15, 22.5 and 30 g/100 g of starch), and gelatinized (90 °C, 20 min). Optical microscopy of the gelatinized starch dispersions (GSDx; x = Tween 80 concentration) revealed that the microstructure was characterized by a continuous phase of leached amylose and amylopectin entangled chains, and a dispersed phase of insoluble remnants, called ghosts, on whose surface small granules were observed, imputed to Tween 80. The apparent viscosity of the GSDx decreased as the concentration of Tween 80 increased (up to about 70-90%). FTIR analysis of dried GSDx indicated that Tween 80 addition decreased short-range ordering. The content of rapidly digestible starch (RDS) and resistant starch (RS) fractions tended to increase significantly, at the expense of a significant decrease of slowly digestible starch (SDS) fraction, an effect that may be attributed to the increase of amorphous structures and starch chain-surfactant complexes. The RDS and RS increase was more pronounced for normal than for waxy corn starch, and the significance of the increase was dependent on Tween 80 concentration. Overall, the results showed that surfactant can affect largely the digestibility of starch chains.

Ultrasound assisted enzymatic hydrolysis for isolating titanium dioxide nanoparticles from bivalve mollusk before sp-ICP-MS.

Applicability of single-particle inductively coupled plasma mass spectrometry (sp-ICP-MS) using dwell times equal to or shorter than 100 µs has been tested for assessing titanium dioxide nanoparticles (TiO2 NPs) in bivalve mollusks. TiO2 NPs isolation from fresh mollusk tissues was achieved by ultrasound assisted enzymatic hydrolysis procedure using a pancreatin/lipase mixture. Optimum extraction conditions imply ultrasonication (60% amplitude) for 10 min, and 7.5 mL of a solution containing 3.0 g L-1of pancreatin and lipase (pH 7.4). The developed method was found to be repeatable (repeatability of 17% for the over-all procedure, TiO2 NPs concentration of 5.33 × 107 ±â€¯8.89 × 106, n = 11), showing a limit of detection of 5.28 × 106 NPs g-1, and a limit of detection in size of 24.4-30.4 nm, based on the 3σ criteria, and on the 3σ/5 σ criteria, respectively. The analytical recovery within the 90-99% range (use of TiO2 NPs standards of 50 nm at 7 and 14 µg L-1 as Ti). Several bivalve mollusks (clams, cockles, mussels, razor clams, oysters and variegated scallops) were analyzed for total titanium (ICP-MS after microwave assisted acid digestion), and for TiO2 NPs by the proposed method. TiO2 NPs concentrations were within the 2.36 × 107-1.25 × 108 NPs g-1 range, and the most frequent sizes were from 50 to 70 nm.

Evaluation of the digestibility of solid lipid nanoparticles of glyceryl dibehenate produced by two techniques: Ultrasonication and spray-flash evaporation.

OBJECTIVE: To evaluate the digestibility of Solid Lipid Nanoparticles (SLN) of glyceryl dibehenate prepared either with surfactants by ultrasonication or without surfactant by spray-flash evaporation. METHODS: SLN of glyceryl dibehenate (Compritol® 888 ATO) were produced by two processes: (i) high-shear homogenization with a solution of water-soluble surfactants followed by ultrasonication (ii) and Spray-Flash Evaporation (SFE) of the pure lipid. The digestibility of these nanoparticles was then tested by in vitro lipolysis using a pH-stat apparatus and the assay of glycerides by gel phase chromatography. RESULTS: SLN of glyceryl dibehenate prepared by ultrasonication exhibited a mean particle size of 180nm and showed a limited digestion of the lipid excipient. SLN comprising only glyceryl dibehenate produced by SFE have a mean particle size between 235 and 411nm depending on process parameters. These nanoparticles were not digested by lipases. The presence of surfactant at the lipid/water interface of the SLN seems to be mandatory to allow the adsorption of the lipase and degradation of glyceryl behenate. CONCLUSIONS: Glyceryl dibehenate as a solid particle - even as a SLN - is not digested by pancreatin during in vitro lipolysis test.

Peptides derived from in vitro gastrointestinal digestion of germinated soybean proteins inhibit human colon cancer cells proliferation and inflammation.

The aim was to investigate the potential of germinated soybean proteins asa source of peptides with anticancer and anti-inflammatory activities produced after simulated gastrointestinal digestion. Protein concentrate from germinated soybean was hydrolysed with pepsin/pancreatin and fractionated by ultrafiltration. Whole digest and fractions>10, 5-10, and<5kDa caused cytotoxicity to Caco-2, HT-29, HCT-116 human colon cancer cells, and reduced inflammatory response caused by lipopolysaccharide in macrophages RAW 264.7. Antiproliferative and anti-inflammatory effects were generally higher in 5-10kDa fractions. This fraction was further purified by semi-preparative chromatography and characterised by HPLC-MS/MS. The most potent fraction was mainly composed of ß-conglycinin and glycinin fragments rich in glutamine. This is the first report on the anti-cancer and anti-inflammatory effects of newly isolated and identified peptides from germinated soybean released during gastrointestinal digestion. These findings highlight the potential of germination as a process to obtain functional foods or nutraceuticals for colon cancer prevention.