triolein and tricaprylin

Oil-soluble vitamins are often encapsulated within emulsion-based delivery systems to facilitate their incorporation into aqueous-based products. We have examined the influence of carrier oil type and simulated small intestinal fluid (SSIF) composition on the bioaccessibility of emulsified vitamin E using a gastrointestinal model. Oil-in-water emulsions containing vitamin E acetate were prepared using bile salts as emulsifier, and either long chain triacylglycerols (glyceryl trioleate, LCT) or medium chain triacylglycerols (glyceryl trioctanoate, MCT) as carrier oils. The addition of calcium (CaCl₂) to the SSIF increased the extent of lipid digestion in LCT-emulsions, but had little impact in MCT-emulsions. The bioaccessibility of vitamin E increased in the presence of calcium and phospholipids (DOPC) in LCT-emulsions, but decreased in MCT-emulsions. The highest bioaccessibility (≈66%) was achieved for LCT-emulsions when the SSIF contained both calcium and phospholipids. The conversion of α-tocopherol acetate to α-tocopherol after in vitro digestion was considerably higher for LCT-emulsions when calcium ions were present in the SSIF, but was not strongly affected by SSIF composition for MCT-emulsions. In general, this research provides important information about the factors influencing the bioaccessibility of emulsified vitamin E, which could be used to design more effective emulsion-based delivery systems for increasing the oral bioavailability of this important bioactive component.

Topics: alpha-Tocopherol; Animals; Antioxidants; Calcium Chloride; Caprylates; Deoxycholic Acid; Digestion; Emulsifying Agents; Emulsions; Food Additives; Food Technology; Food, Fortified; Humans; Hydrolysis; Intestinal Secretions; Models, Biological; Nutritive Value; Sodium Cholate; Solubility; Triglycerides; Triolein

Despite the considerable number of in vivo and in vitro studies on the digestive fate of lipophilic nutrients, micronutrients, and bioactives, the effects of the structure and composition of foods on the physicochemical mechanisms of luminal digestion are still poorly understood. Studying them is indeed complex because the number of parameters is high and many of them are interdependent. To solve this problem, an in silico simulation based on a multi-agent system was recently proposed to study the intestinal bioaccessibility of lipophilic nutrients and micronutrients from a single oil droplet. The roles of lipolysis and solubilization in bile salt were included. The effects of several food and digestion parameters were in line with those reported in the experimental literature. The goal of the research reported in this new article was to include more digestion parameters in the simulation in order to make it more realistic against complex cases. This was done in one specific digestion condition reflecting in vitro experiments, using droplets of tricaprylin or triolein containing vitamin A. The structure and principles of the original model were kept, with independent local modifications in order to study each factor separately. First, a gastric step was added where lipolysis took place, and only a marginal effect on the following intestinal step was found. Then, the chemical form of vitamin A, either non-hydrolyzed retinyl ester or retinyl ester instantly hydrolyzed into retinol, was investigated by considering different localizations in the droplet, resulting in a higher bioaccessibility for the retinol. The case of a mixture of tricaprylin and triolein indicated an influence of the oil phase viscosity. The consideration of mixed micelles compared to simple bile salt micelles was also investigated, and resulted in a higher vitamin A bioaccessibility, especially with triolein. Finally, a full model including the most influential parameters was tested to simulate the digestion of triglyceride-limonene mixtures, giving bioaccessibility trends in very good agreement with the literature.

Topics: Animals; Bile Acids and Salts; Caprylates; Chemical Phenomena; Computational Biology; Computer Simulation; Dietary Fats; Digestion; Expert Systems; Food Analysis; Gastric Juice; Humans; Hydrophobic and Hydrophilic Interactions; Intestinal Absorption; Lipolysis; Models, Biological; Solubility; Triglycerides; Triolein; Viscosity; Vitamin A

Second generation lipid systems for the delivery of bioactive compounds have been developed by mixing a liquid carrier oil with a solid lipid to form so-called nanostructured lipid carriers (NLCs). In this study, we investigated the effect of different liquid carrier oils on the crystallization and aggregation behavior of tristearin NLC dispersions. We found that NLC suspension stability was strongly affected by the type and amount of the carrier oil. As the oil concentration was increased, the crystallization and melting temperatures decreased, the polymorphic transformation rate increased, the particles became more spherical, and suspension stability was enhanced. These results suggest that oil trapped within the growing crystal matrix accelerated polymorphic transformation but retarded the large shape change normally associated with the transformation. We also found that considerably less surfactant was necessary to produce stable NLC suspensions than was required to stabilize solid lipid nanoparticle (SLN) suspensions without a carrier oil. Based on preliminary simulation results, we hypothesized that improved NLC suspension stability was attributable to both reduced particle shape change, which created less new surface area to be covered by surfactant, and increased mobility of surfactant molecules, which resulted in available surfactant being more efficient at covering created surface area.

Topics: Alkanes; Calorimetry, Differential Scanning; Caprylates; Crystallization; Drug Carriers; Drug Stability; Flocculation; Microscopy, Electron, Transmission; Nanostructures; Oleic Acid; Olive Oil; Palm Oil; Particle Size; Phase Transition; Plant Oils; Polysorbates; Surface-Active Agents; Suspensions; Triglycerides; Triolein

Efficient dietary fat digestion is essential for newborns who consume more dietary fat per body weight than at any other time of life. In many mammalian newborns, pancreatic lipase related protein 2 (PLRP2) is the predominant duodenal lipase. Pigs may be an exception since PLRP2 expression has been documented in the intestine but not in the pancreas. Because of the differences in tissue-specific expression, we hypothesized that the kinetic properties of porcine PLRP2 would differ from those of other mammals. To characterize its properties, recombinant porcine PLRP2 was expressed in HEK293T cells and purified to homogeneity. Porcine PLRP2 had activity against tributyrin, trioctanoin and triolein. The activity was not inhibited by bile salts and colipase, which is required for the activity of pancreatic triglyceride lipase (PTL), minimally stimulated PLRP2 activity. Similar to PLRP2 from other species, PLRP2 from pigs had activity against galactolipids and phospholipids. Importantly, porcine PLRP2 hydrolyzed a variety of dietary substrates including pasteurized human mother's milk and infant formula and its activity was comparable to that of PTL. In conclusion, porcine PLRP2 has broad substrate specificity and has high triglyceride lipase activity even in the absence of colipase. The data suggest that porcine PLRP2 would be a suitable lipase for inclusion in recombinant preparations for pancreatic enzyme replacement therapy.

Topics: Amino Acid Sequence; Animals; Blotting, Western; Caprylates; Cattle; Colipases; Dietary Fats; Galactolipids; Humans; Hydrolysis; Intestinal Mucosa; Lipase; Molecular Sequence Data; Pancreas; Phospholipids; Real-Time Polymerase Chain Reaction; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sequence Homology, Amino Acid; Substrate Specificity; Swine; Triglycerides; Triolein

In a previous study, we demonstrated that the beta5'-loop in the C-terminal domain of human pancreatic triglyceride lipase (hPTL) makes a major contribution in the function of hPTL (Chahinian et al. (2002) Biochemistry 41, 13725-13735). In the present study, we characterized the contribution of three residues in the beta5'-loop, Val-407, Ile-408, and Leu-412, to the function of hPTL. By substituting charged residues, aspartate or lysine, in these positions, we altered the hydrophilic to lipophilic ratio of the beta5'-loop. Each of the mutants was expressed, purified, and characterized for activity and binding with both monolayers and emulsions and for binding to colipase. Experiments with monolayers and with emulsions suggested that the interaction of hPTL with a phospholipid monolayer differs from the interaction of the hPTL-colipase complex with a dicaprin monolayer or a triglyceride emulsion (i.e. neutral lipids). Val-407, Ile-408, and Leu-412 make major contributions to interactions with monolayers, whereas only Val-407 and Ile-408 appear essential for activity on triglyceride emulsions in the presence of bile salt micelles. In solutions of taurodeoxycholate at micellar concentrations, a major effect of the beta5'-loop mutations is to change the interaction between hPTL and colipase. These observations support a major contribution of residues in the beta5'-loop in the function of hPTL and suggest that a third partner, bile salt micelles or the lipid interface or both, influence the binding of colipase and hPTL through interactions with the beta5'-loop.

Topics: Adsorption; Aspartic Acid; Bile Acids and Salts; Caprylates; Colipases; Diglycerides; DNA, Complementary; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Humans; Isoleucine; Kinetics; Leucine; Lipase; Lysine; Micelles; Models, Molecular; Mutagenesis, Site-Directed; Mutation; Pancreas; Phosphatidylcholines; Phospholipids; Pressure; Protein Binding; Protein Conformation; Protein Interaction Mapping; Protein Structure, Secondary; Proteins; Time Factors; Triglycerides; Triolein; Valine

Progenipoietin (ProGP), a dual receptor agonist of fetal liver tyrosine kinase-3 (flt3) and granulocyte colony-stimulating factor (G-CSF) receptors, has been shown to significantly enhance production of both polymorphonuclear leukocytes and dendritic cells (DCs) in the peripheral blood and spleen of mice, when administered as daily s.c. injections for about 10 days. Here, we have successfully designed a sustained-delivery formulation for this novel chimeric protein using multivesicular liposomes (DepoFoam), and studied the effects of changing both the triglyceride and phospholipid composition of the lipid matrix to modulate its delivery profile. Encapsulation of ProGP in these particles led to retention of its structural integrity, and maintenance of its biological activity in vivo. Administration of a single s.c. dose of 1mg/kg of an optimized DepoProGP formulation in rats, led to significant elevation of absolute neutrophil counts (ANC) that were maintained at levels >10,000 microliter(-1) for 9-11 days, in a reproducible manner. In contrast, administration of the unencapsulated ProGP at the same dose, resulted in elevation of neutrophils by day 1, followed by a quick decline to base line levels by day 3. These data suggest the possibility of administering a single dose of DepoFoam-encapsulated ProGP to improve hematopoietic recovery time after chemotherapy, and for other indications that require multiple daily doses of ProGP.

Topics: Animals; Caprylates; Chromatography, High Pressure Liquid; Colony-Stimulating Factors; Delayed-Action Preparations; Drug Stability; Humans; Injections, Subcutaneous; Leukocyte Count; Liposomes; Male; Neutrophils; Particle Size; Phosphatidylcholines; Phosphorylcholine; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Solubility; Surface Properties; Triglycerides; Triolein

The ability of water-insoluble molecules such as triacylglycerols to partition from oil phases into phospholipid interfaces may be crucial to their hydrolysis by lipases in the aqueous environment of plasma and cells. This study uses high resolution and magic angle spinning 13C NMR spectros-copy to measure the solubility of the 8-carbon medium chain triacylglycerol, trioctanoin, in the lamellar structure of phospholipids (vesicles and multilayers) in the presence of other neutral lipids that may compete for an interfacial location (long chain triacylglycerol, cholesteryl ester, and cholesterol). In the presence of a saturating concentration of triolein (approximately 3 mole%), the solubility of trioctanoin in egg phosphatidylcholine vesicles decreased from 10 mole% to 7 mole%. The presence of a saturating concentration of trioctanoin (approximately 10 mole%) decreased the interfacial solubility of long chain triolein to approximately 1 mole%. Cholesteryl oleate in phospholipid vesicles slightly diminished the incorporation of trioctanoin into the surface. The presence of cholesterol reduced the interfacial solubility of trioctanoin, but at a high level of cholesterol (30 mole%), trioctanoin had a solubility of 3 mole%. Thus, even in the presence of other competing neutral lipids, medium chain triacylglycerol retains a favorable location and surface concentration for efficient hydrolysis. 13C NMR analysis thus provides an explanation for preferential hydrolysis of medium, compared to long chain triacylglycerol, in a physical blend of medium and long chain triacylglycerol in a single emulsion particle, and in general, a valuable approach to determine substrate availability at phospholipid surfaces.

Topics: Caprylates; Cholesterol; Cholesterol Esters; Emulsions; Hydrolysis; In Vitro Techniques; Lipase; Lipid Bilayers; Liposomes; Magnetic Resonance Spectroscopy; Phospholipids; Solubility; Triglycerides; Triolein; Water

The coding sequences of the Rhizopus delemar lipase and prolipase were altered by oligonucleotide-directed mutagenesis to introduce amino acid substitutions. The resulting mutant enzymes, synthesized by the bacterial host Escherichia coli BL21 (DE3), were tested for their ability to hydrolyze the triglycerides triolein (TO), tricaprylin (TC) and tributyrin (TB). Mutagenesis and lipase gene expression were carried out using plasmid vectors derived from previously described recombinant plasmids [Joerger and Haas (1993) Lipids 28, 81-88] by introduction of the origin of replication of bacteriophage f1. Substitution of threonine 83 (thr83), a residue thought to be involved in oxyanion binding, by alanine essentially eliminated lipolytic activity toward all substrates examined (TB, TO and TC). Replacement of thr83 with serine caused from two- to sevenfold reductions in the activity toward these substrates. Introduction of tryptophan (trp) at position 89, where such a residue is found in closely related fungal lipases, reduced the specific activity toward the three triglyceride substrates. For the mutagenesis of residues in the predicted acyl chain binding groove, mutagenic primers were designed to cause the replacement of a specific codon within the prolipase gene with codons for all other amino acids. Phenylalanine 95 (phe95), phe112, valine 206 (val206) and val209, were targeted. A phenotypic screen was successfully employed to identify cells producing prolipase with altered preference for olive oil, TC or TB. In assays involving equimolar mixtures of the three triglycerides, a prolipase with a phe95-->aspartate mutation showed an almost twofold increase in the relative activity toward TC. Substitution of trp for phe112 caused an almost threefold decrease in the relative preference for TC, but elevated relative TB hydrolysis. Replacement of val209 with trp resulted in an enzyme with a two- and fourfold enhanced preference for TC and TB, respectively.

Topics: Base Sequence; Binding Sites; Caprylates; Codon; Escherichia coli; Gene Expression; Hydrolysis; Lipase; Molecular Sequence Data; Molecular Structure; Mutagenesis, Site-Directed; Olive Oil; Plant Oils; Plasmids; Recombinant Proteins; Rhizopus; Structure-Activity Relationship; Substrate Specificity; Triglycerides; Triolein

In the present study, porcine pancreatic lipase, rabbit gastric lipase, and human gastric lipase stereospecificity toward chemically alike, but sterically nonequivalent ester groups within one single triglyceride molecule was investigated. Lipolysis reactions were carried out on synthetic trioctanoin or triolein, which are homogenous, prochiral triglycerides, chosen as models for physiological lipase substrates. Diglyceride mixtures resulting from lipolysis were derivatized with optically active R-(+)-1-phenylethylisocyanate, to give diastereomeric carbamate mixtures, which were further separated by high performance liquid chromatography. Resolution of diastereomeric carbamates gave enantiomeric excess values, which reflect the lipases stereobias and clearly demonstrate the existence of a stereopreference by both gastric lipases for the sn-3 position. The stereoselectivity of human and rabbit gastric lipases, expressed as the enantiomeric excess percentage, was 54% and 70% for trioctanoin and 74% and 47% for triolein, respectively. The corresponding values with porcine pancreatic lipase were 3% in the case of trioctanoin and 8% in that of triolein. It is worth noting that rabbit gastric lipase, unlike human gastric lipase, became more stereoselective for the triglyceride with shorter acyl chains (trioctanoin). This is one of the most striking catalytic differences observed between these two gastric lipases.

Topics: Animals; Caprylates; Carbamates; Gastric Juice; Humans; Lipase; Lipolysis; Pancreas; Rabbits; Stereoisomerism; Substrate Specificity; Swine; Triglycerides; Triolein

Fifteen unsuckled neonatal piglets from three litters were fasted for 2 h after birth then allotted within litter to a 2 x 3 factorial arrangement of treatments and given by stomach tube 2.36, 4.91 or 11.32 g of either (1-14C)-triolein or (1-14C)-trioctanoin/BW (kg).75. The hourly 14CO2 production was measured for 12 h for estimating the oxidation rate of dosed triacylglycerols. The contents of each segment of the digestive tract were collected for 14C and lipid analyses for calculating absorption of the dosed triacylglycerols. Grams of trioctanoin absorbed (P less than .01) and oxidized (P less than .001) by piglets were higher than grams of triolein. The maximum amounts of trioctanoin and triolein absorbed in 12 h were 2.77 +/- .29 and 1.83 +/- .36 g/BW (kg).75, respectively. The maximum amounts of trioctanoin and triolein oxidized in 12 h were 1.60 +/- .14 and .33 +/- .17 g/BW (kg).75, respectively. The maximum amounts of trioctanoin and triolein oxidization during 1-h periods were .231 +/- .017 and .049 +/- .021 g/BW (kg).75, respectively. The calculated maximum percentage of piglets' daily maintenance energy needs supplied by trioctanoin oxidation was 42.0%, compared with 10.6% from triolein. The maximum utilization of trioctanoin occurred at the intermediate dose. Results indicated that trioctanoin was used more effectively than triolein; it may be useful as a source of supplemental energy to improve the survival of piglets. A single dose should provide about 6 g/body wt (kg).75.

Topics: Animals; Animals, Newborn; Caprylates; Digestion; Intestinal Absorption; Oleic Acid; Oleic Acids; Oxidation-Reduction; Swine; Triglycerides; Triolein

Three substrates labeled with nonradioactive 13C have been employed to establish a trilogy of noninvasive breath tests to detect fat malabsorption in children and then to differentiate the etiology of the steatorrhea. Administration of 17 mg/kg of (13C)triolein Lipomul (The Upjohn Co., Kalamazoo, Mich.) resulted in a peak excretion rate of 13CO2 greater than 2.7% dose/h in 10 normal subjects (mean value 4.96 +/- 2.2% dose/h) whereas all 17 subjects with fat malabsorption were below this value (mean value, 0.75% +/- 0.63% dose/h); p less than 0.001). For the detection of fat malabsorption, the discriminative value of (13C)triolein was superior, 100% sensitive, and 89% specific, while the use of (13C)palmitic acid (17 mg/kg) or (13C)trioctanoin (7.5 mg/kg) alone yielded both false-positive and false-negative results. In 6 out of 6 cases, pancreatic insufficiency could be differentiated from mucosal disease (7 patients) or bile salt deficiency (4 patients) by the presence of abnormal triolein or trioctanoin breath tests, or both but normal palmitic acid breath tests. However, further differentiation of mucosal disorders from bile salt disorders could not be achieved using either a single- or a multiple-substrate breath test. The use of the single triolein breath test in children offers an attractive, sensitive alternative to conventional fecal fat measurements to establish the presence of steatorrhea, and when using 12C-lipid with multiple substrates, the tests are capable of providing additional insight into the mechanism of fat malabsorption.

Topics: Adolescent; Biliary Tract Diseases; Breath Tests; Caprylates; Carbon Isotopes; Celiac Disease; Child; Child, Preschool; Exocrine Pancreatic Insufficiency; Humans; Infant; Intestinal Diseases; Palmitic Acid; Palmitic Acids; Triglycerides; Triolein