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The formation and permeability of drugs across free pectin and chitosan films prepared by a spraying method.

This study has investigated the permeation of drugs through free films made of pectin and chitosan. The background for this study is the intended use of the films as coating material in a colon-specific drug delivery device. The factors that varied when making the films were the pectin source and grade of the pectin, degree of deacetylation of the chitosan and ratio between pectin and chitosan. The permeability of the model drug in 0.1 M HCl was low with an average drug release of 1.3 x 10(-3)%/cm. The films containing high content of chitosan showed exponential kinetics while the films containing high content of pectin showed 0-order kinetics. The release of drug in phosphate buffer pH 6.8 showed 0-order kinetics. The lowest permeability was obtained for a film consisting of a high content of pectin to chitosan, chitosan with a high degree of deacetylation and non-amidated low methoxylated citrus pectin. The permeation of paracetamol for this combination was 9.4 x 10(3)%/cm. This film combination had a combined diffusion of only 0.046%/cm after 1 h in 0.1 M HCl and 4 h in phosphate buffer pH 6.8.

Chitin↗

Effects of calcium, pH, and blockiness on kinetic rheological behavior and microstructure of HM pectin gels.

The kinetic behavior during gel formation and the microstructure of 0.75% high methoxyl (HM) pectin gels in 60% sucrose have been investigated by oscillatory measurements and transmission electron microscopy for three comparable citrus pectin samples differing in their degree of blockiness (DB). Ca2+ addition at pH 3.0 resulted in faster gel formation and a lower storage modulus after 3 h for gels of the blockwise pectin A. For gels of the randomly esterified pectin B, the Ca2+ addition resulted in faster gel formation and a higher storage modulus at pH 3.0. At pH 3.5, both pectins A and B were reinforced by the addition of Ca2+. In the absence of Ca2+, the shortest gelation time was obtained for the sample with the highest DB. Microstructural characterization of the gel network, 4 and 20 h after gel preparation, showed no visible changes on a nanometer scale. The microstructure of pectins A and B without Ca2+ was similar, whereas the presence of Ca2+ in pectin A resulted in an inhomogeneous structure.

Calcium↗

Deposition of pectin/poly-L-lysine multilayers with pectins of varying degrees of esterification.

The effect of pectin esterification on the assembly of multilayers consisting of poly-L-lysine (PLL) and pectin was studied using surface plasmon resonance (SPR), Fourier transform infrared-attenuated total reflection spectroscopy (FTIR-ATR), and a quartz crystal microbalance with dissipation monitoring (QCMD). With each layer deposited, there was a progressive increase in mass. The net charge of the multilayers was positive and increased with increasing degree of esterification of the pectin. Multilayer fabrication involved a limited fractionation of the pectin preparations, with the more highly esterified pectins having a weaker affinity for PLL. The multilayers were relatively hydrated structures with estimates of solids content in the range 10-32% w/w. The more highly esterified pectins had a tendency to form more hydrated structures, which showed a strong deswelling when PLL was added to a freshly deposited pectin layer.

Esterification↗

Isothermal titration calorimetry study of pectin-ionic surfactant interactions.

Isothermal titration calorimetry (ITC) was used to measure enthalpy changes resulting from injection of anionic (sodium dodecyl sulfate, SDS) or cationic (dodecyl trimethylammonium bromide, DTAB) surfactants into aqueous 1 wt % pectin solutions (30, 60, or 90% methoxylated). In the absence of pectin, the critical micelle concentrations (cmc) determined by ITC were 14.7 mM for DTAB and 7.7 mM for SDS. Binding of DTAB to pectin was endothermic and was attributed to electrostatic attraction between the cationic surfactant and anionic biopolymer. Binding of SDS to pectin was exothermic and was attributed to hydrophobic interactions. Pectin reduced the cmc of SDS, probably because of long-range electrostatic repulsion between the molecules. Above a particular concentration, which depended on pectin and surfactant type, both ionic surfactants promoted pectin aggregation (monitored by turbidity increase). This study demonstrates the potential of ITC for providing valuable information about interactions between polysaccharides and amphiphiles.

Calorimetry↗

Particle size distribution of orange juice cloud after addition of sensitized pectin.

The effect of the addition of commercial pectin to orange juice that was de-esterified by alkaline treatment or pectinmethylesterase was evaluated. Pectin at various degrees of esterification (DE) was added to reconstituted frozen orange juice concentrate, and the extent of cloud loss was determined after centrifugation. Of the five pectin treatments, percent transmittance (% T) of juice remained <5%, except for the pectin treatment with a residual methoxyl content (% DE) of <5%. Addition of 100--600 ppm of 5% DE pectin to juice resulted in % T between 36 and 73%. Despite the absence of clarification, increases in cloud particle size were observed and depended on the % DE, the amount of pectin added, and the method used to modify the pectins.

Beverages↗

Solvent effects on the molecular properties of pectins.

Measurements revealed that LiAc/HAc buffer, when compared with other solvents, gave relatively low values of turbidity for five commercial pectins with various apparent molar masses and degrees of methyl esterification (DE). Therefore, HPSEC with on-line light scattering and viscosity detection was employed to compare LiAc/HAc buffer against NaNO(3) solution, a commonly used mobile phase for measuring the molecular properties of these pectins and an additional pectin prepared by microwave extraction. Microwave-extracted pectin was included in the study for its higher molar mass and DE compared with commercial pectins. Most commercial samples were more soluble and had a higher molar mass when dissolved in NaNO(3) than in LiAc/HAc buffer, whereas the microwave-extracted pectin was more soluble in LiAc/HAc buffer and had about the same molar mass. Furthermore, association fragments of pectin contained in samples were more dissociated by LiAc/HAc buffer than by NaNO(3). For the samples studied, weight-average molar masses ranged from about 41000 to 307000, weight-average intrinsic viscosities from about 0.86 to 9.76 dL/g, z-average radii of gyration from about 13 to 45 nm, and Mark-Houwink constants from about 0.62 to 0.94.

Chromatography↗

New method for a two-step hydrolysis and chromatographic analysis of pectin neutral sugar chains.

A new method for the determination of the main neutral sugars in pectin has been developed. The sample preparation involves a mild chemical attack followed by an enzymatic hydrolysis. The completeness and nondestructive character of the method are demonstrated by comparison of the results obtained with different acids such as H2SO4, HCl, and trifluoroacetic acid (TFA) at different concentrations (2, 1, or 0.2 M) at two temperatures (80 or 100 degrees C). The chemical hydrolysis of pectin neutral sugar chains with strong acid (1 or 2 M) and high temperature (100 degrees C) shows that the liberation of the pectin sugars is not realized at the same rate for each sugar. Different optimum conditions are thus obtained. However, the chemical pectin hydrolysis with 0.2 M TFA at 80 degrees C is characterized by the liberation of pectin neutral sugar side chains without any degradation within 72 h of hydrolysis. Under these conditions, the liberation of some pectin sugars, essentially galactose, glucose, and rhamnose, was not complete. An enzymatic hydrolysis is necessary to obtain a complete release of all the sugars. The combination of the two treatments, a chemical hydrolysis realized with diluted acid (0.2 M) for 72 h at low temperature (80 degrees C) on one hand and an enzymatic hydrolysis on the other hand, allow a total liberation of pectin sugars. The quantitative analysis of the carbohydrates is realized with accuracy, high selectivity, and sensitivity with high-performance anion-exchange chromatography with pulsed-amperometric detection. The sugars can be analyzed without any derivatization with a limit of quantification of 0.1 mM.

Acids↗

Cellulase and protease preparations can extract pectins from various plant byproducts.

The use of protease and cellulase preparations to extract pectins from plant byproducts (chicory, cauliflower) was investigated. Different enzymatic preparations were characterized by their activities toward proteins, cellulose, and pectins. These preparations were then tested regarding pectin extraction, and extraction conditions (nature and concentration of enzyme, incubation time) were optimized. Enzymatic and acidic extractions were compared and also combined in sequential extractions. This study shows that it is possible to extract pectins by using cellulases and proteases. Enzymes can extract pectins with a higher yield ( approximately 35%) than acid (approximately 28%) but enzyme-extracted pectins have a smaller molar mass (300,000 g/mol) than acid-extracted pectins (500,000 g/ mol). Different hypotheses are tested and discussed to explain this mass difference.

Brassica↗

Impact of electrostatic interactions on formation and stability of emulsions containing oil droplets coated by beta-lactoglobulin-pectin complexes.

Interfacial protein-polysaccharide complexes can be used to improve the physical stability of oil-in-water emulsions. The purpose of this study was to examine the impact of ionic strength on the formation and stability of oil-in-water emulsions containing polysaccharide-protein-coated droplets. Emulsions were prepared that contained 0.1 wt % corn oil, 0.05 wt % beta-lactoglobulin, and 0.02 wt % pectin at pH 7. The emulsions were then adjusted to pH 4 to promote electrostatic deposition of the pectin molecules onto the surfaces of the protein-coated droplets. The salt concentration of the aqueous phase (0 or 50 mM NaCl) was adjusted either before or after deposition of the pectin molecules onto the droplet surfaces. We found that stable emulsions containing polysaccharide-protein-coated droplets could be formed when the salt was added after pectin adsorption but not when it was added before pectin adsorption. This phenomenon was attributed to the ability of NaCl to promote droplet flocculation in the protein-coated droplets so that the pectin molecules adsorbed onto the surfaces of flocs rather than individual droplets when salt was added before pectin adsorption. We also found that polysaccharide-protein-coated droplets had a much improved stability to salt-induced flocculation than protein-coated droplets with the same droplet charge (zeta-potential). Theoretical predictions indicated that this was due to the ability of the adsorbed polysaccharide layer to strongly diminish the van der Waals attraction between the droplets.

Adsorption↗

Comparison of the in vitro bifidogenic properties of pectins and pectic-oligosaccharides.

AIMS: To compare the in vitro fermentation properties of pectins and oligosaccharides derived from them in pure and mixed faecal cultures. METHODS AND RESULTS: Specific growth rates of selected bacterial genera were calculated in pure culture. Bifidobacterium angulatum, B. infantis and B. adolescentis had higher growth rates on pectic oligosaccharides (POS I) derived from high methylated pectin (HMP) than on HMP and B. pseudolongum and B. adolescentis on pectic oligosaccharides (POS II) derived from low methylated pectin than on HMP. Controlled pH batch mixed faecal cultures were then carried out and a prebiotic index was calculated as a mean to compare the fermentation properties of the different substrates. In general, greater fermentation selectivity was obtained with lower degrees of methylation (PI24(-HMP) = -0.11, PI24(-LMP) = 0.033; PI24(-POS I) = 0.071 and PI24(-POS II) = 0.092). An effect of size on prebiotic potential was observed, with the oligosaccharides having more selective fermentation properties than the pectins they derived from. CONCLUSIONS: The degree of methylation plays an important role in the fermentation properties of pectins. Pectic-oligosaccharides are a better prebiotic candidate than the pectins, although their bifidogenic effect is low compared to oligofructose. SIGNIFICANCE AND IMPACT OF THE STUDY: The effect of size on prebiotic potential was demonstrated. Non-selectively fermented polysaccharides like pectin can have their bifidogenic properties improved by partial hydrolysis.

Bacteria↗

Characterization of pectin methylesterase B, an outer membrane lipoprotein of Erwinia chrysanthemi 3937.

The secretion of extracellular pectinases, among which there are least six isoenzymes of pectate lyase and one pectin methylesterase, allows the phytopathogenic bacterium Erwinia chrysanthemi to degrade pectin. A gene coding for a novel pectin methylesterase has been cloned from an E. chrysanthemi strain 3937 gene library. This gene, pemB, codes for a 433-amino-acid protein. The PemB N-terminal region has the characteristics of lipoprotein signal sequences. We have shown that the PemB precursor is processed and that palmitate is incorporated into the mature protein. The PemB lipoprotein is not released into the extracellular medium and is localized in the outer membrane. The PemB sequence presents homology with other pectin methylesterases from bacterial and plant origin. pemB-like proteins were detected in four other E. chrysanthemi strains but not in Erwinia carotovora strains. PemB was overproduced in Escherichia coli and purified to homogeneity. PemB activity is strongly increased by non-ionic detergents. The enzyme is more active on methylated oligogalacturonides than on pectin, and it is necessary for the growth of the bacteria on oligomeric substrates. PemB is more probably involved in the degradation of methylated oligogalacturonides present in the periplasm of the bacteria, rather than in a direct action on extracellular pectin. pemB expression is inducible in the presence of pectin and is controlled by the negative regulator KdgR.

Amino Acid Sequence↗

Effect of wheat bran and pectin on the absorption and retention of phosphorus, calcium, magnesium and zinc by the growing pig.

Two separate balance experiments of P, Ca, Mg and Zn were carried out on 5 lots of 4 growing pigs each (35-40 kg) adapted for 3 weeks to one of the diets studied. In the first experiment, the control diet was compared with a diet containing 20% of coarse wheat bran, thus richer in minerals, the quantities ingested not being equalized. In the second experiment, three diets were compared: a control diet, a diet with 2.5% of high-methoxylated (HM) apple pectin, and a diet with 2.5% low-methoxylated (LM) apple pectin. The supplement of P and Mg provided by the wheat bran was well absorbed (apparent absorption) and retained by the pigs. On the contrary, in spite of higher intake of Ca and Zn with bran diet, the absorption of these minerals was not improved. The action of wheat bran phytase and the possible absorption of P and Mg (but not of Ca and Zn) in the large intestine could explain these results. Compared to HM pectin that had relatively little effect on mineral utilization, LM pectin drastically diminished the absorption and retention of the minerals studied and resulted in negative Ca, Mg and Zn balances. The degree of pectin esterification would thus be the main factor determining the effect of pectin on mineral availability. In conclusion, wheat bran is a source of available P and Mg for the pig but it might have an unfavorable effect on the utilization of Ca and Zn. LM pectin produces a deleterious influence on mineral balances.

Absorption↗

The effect of wheat bran and pectin on paracetamol absorption in the rat.

1. The absorption of an oral dose of paracetamol was measured in rats given either a basal fibre-free diet, or the basal diet with either pectin or bran added. 2. Urinary excretion of the oral dose was significantly greater during the first 8 h in the pectin-fed rats compared with those on basal diet, though cumulative excretion after 72 h was the same. 3. Free paracetamol levels in the plasma were significantly higher in the pectin-fed rats compared with those on basal diet at 30, 60, 90 and 120 min after the oral dose. 4. The plasma half-life of intravenously-injected paracetamol was shorter in the pectin-fed rats than in those on basal diet alone, but the antipyrine half-lives were not significantly different. 5. Pectin feeding had no effect on either the apparent volume of distribution of paracetamol and antipyrine, or on the rate of gastric empyting. 6. Perfusion of the whole length of the small bowel showed a significantly greater capacity for paracetamol absorption in the pectin-fed rats. 7. Bran had no effect on paracetamol absorption. 8. It was concluded that dietary fibre intake affects drug absorption and that the effect varies with the type of fibre. Unexpectedly pectin accelerates rather than retards absorption of paracetamol, though the mechansim of this effect is unknown.

Acetaminophen↗

Predominant conjugation with glycine of biliary and lumen bile acids in rats fed on pectin.

1. Bile acids were analysed in the bile and lumen samples of rats which received a cholesterol-free or cholesterol-enriched (5 g/kg) diet free from fibre, or containing cellulose or citrus pectin at the level of 100 g/kg. 2. Dietary pectin but not cellulose increased biliary bile acid concentration and excretion. Dietary cholesterol did not affect biliary bile acids quantitatively. 3. Biliary bile acids were almost exclusively conjugated with glycine or taurine in the various experimental situations. The predominant portion of bile acids in rats fed on the cholesterol-free diet was conjugated with taurine when the diet was either free from fibre or contained cellulose; the ratio of bile acids conjugated with glycine: those conjugated with taurine (G:T) was less than 0.2. In contrast, with pectin as a fibre source, the conjugation with glycine increased enormously (G:T increased to approximately 4). Cholesterol enrichment of the diet also increased the glycine conjugation in all groups of rats. Even in this situation, the G:T was highest in rats fed on pectin. 4. Pectin, but not cellulose, increased the bile acid content of the small intestine and caecum, both in rats fed on the cholesterol-free and cholesterol-enriched diets. Cholesterol feeding doubled the bile acid content of the caecum in rats fed on a fibre-free diet or a cellulose diet, but not in those fed on pectin. No such effect of cholesterol was observed in the small intestine, except for the ileal bile acid content in rats fed on cellulose. 5. A considerable portion of the bile acids in the small intestine was deconjugated. The extent of the deconjugation was higher in the ileum than in the jejunum. As in the bile, G:T in rats fed on pectin (3-5.5) were higher than those in the other groups (0.05-1.05) in various situations. Also, cholesterol feeding considerably increased the ratio in all groups of rats. 6. The observed dietary alteration of the partition of bile acids between glycine and taurine may be of physiological significance in regulating bile acid and lipid metabolism in rats.

Animals↗

A study on lipid metabolism in heart and liver of cholesterol- and pectin-fed rats.

Pectin is known as a cholesterol-reducing dietary fibre, and in the present study we addressed the question whether pectin affected the quantity of lipid in droplets in the myocardial cells and of lipid in the liver cells. Male Wistar rats received either a diet containing cholesterol or a standard diet without cholesterol with 0, 50 or 100 g pectin/kg incorporated for 10 d. The fractional volume of lipid droplets in the myocardial cells decreased as a function of pectin dose in both the standard-fed and the cholesterol-fed rats. Serum cholesterol was significantly reduced in both groups after addition of 100 g pectin/kg diet. The cholesterol diet increased the liver cholesterol level, and 100 g pectin/kg diet resulted in a lower concentration of liver cholesterol in the cholesterol-fed animals, but the influence on standard-fed rats was modest. Hydroxymethylglutaryl-CoA reductase (EC 1.1.1.88; HMG-CoA reductase) activity increased when pectin was given in the standard diet. Liver triacylglycerol level increased after cholesterol and pectin feeding. Mitochondrial fatty acid oxidation and phosphatidate phosphohydrolase (EC 3.1.3.4) activity tended to decrease, whereas the peroxisomal fatty acid oxidation and acyl-CoA oxidase activity were unchanged. Increased hepatic triacylglycerol content by cholesterol and pectin treatment may be due to inhibited mitochondrial fatty acid oxidation along with increased availability of fatty acid for esterification and triacylglycerol synthesis. The presence of pectin in the diets of cholesterol-fed rats resulted in increased hepatic concentration of triacylglycerols and increased mitochondrial fatty acid oxidation. In this case the hepatic accumulation of triacylglycerol may be mediated by a reduced efflux of triacylglycerols from the liver.

Animals↗

The influence of 5% and 10% dietary apple pectin on parameters of fermentation in faeces and caecal digesta of weaning pigs.

In order to determine the effect of pectin on fermentation parameters in the faeces and caecal digesta of weaned pigs 18 castrated male crossbred pigs with an average body weight of 8 kg were fitted with T-cannulas at the caecum. The animals were randomly distributed into three groups and fed with diets supplemented with 0, 5 and 10% pectin. Faeces were collected over a period of 3 days. Thereafter the diets were withdrawn for 24 h followed by ad libitum feeding to enhance the feed intake. Caecal chyme was collected 0, 8 and 24 h postprandial. In the faeces the addition of 5% pectin to the diet lowered the content of dry matter and lactic acid. The pH and the digestibility of pectins, the concentration of total SCFA, acetate, propionate, butyrate, bicarbonate and chloride increased. Dietary pectin of 10% increased the content of total SCFA and acetate further. When the diets were withdrawn and fed ad libitum 24 h later, a decline of the pH and an increased concentration of lactate in the caecal chyme could be observed in all groups up to 8 h after feeding. With an interval of 8 to 24 h after feeding, a further decline in pH and a rise of lactate only occurred when the diet was not supplemented with pectin. It was concluded that pectin might be beneficial for the development of fermentative processes in the large intestine.

Animals↗

Oral sustained delivery of theophylline and cimetidine from in situ gelling pectin formulations in rabbits.

The aim of this study was to evaluate the potential of an in situ gelling pectin formulation as a vehicle for the oral sustained delivery of theophylline and cimetidine. In vitro studies demonstrated diffusion-controlled release of theophylline from 1, 1.5, and 2% w/v pectin gels. Release of this drug from 1.5% w/v pectin gels formed in situ in rabbit stomach was sustained over a period of 12 hours giving a theophylline bioavailability some seven fold higher than when administered from a commercial syrup. In contrast, interactions between cimetidine and pectin led to weak gelation of the pectin sols that prevented any meaningful determination of in vitro release characteristics. Similarly, in vivo release profiles from pectin formulations containing cimetidine were similar to that from a solution of this drug in buffer, indicative of weak gelation. Examination of the content of the rabbit stomach 5 hours after administration of 1.5% w/v pectin sols containing drug confirmed gel formation, but gels containing cimetidine were noticeably softer than those containing theophylline.

Administration, Oral↗

Effect of pectin on satiety in healthy US Army adults.

OBJECTIVE: We tested the hypothesis that pectin, a source of dietary fiber that delays gastric emptying, increases satiety. METHODS: Male (n = 49) and female (n = 25) US Army employees within normal weight limits were fasted overnight then fed 448 mL of orange juice on 2 separate days followed 4 hours later by 0.473 L of ice cream. On 1 of the 2 days, 5, 10, 15 or 20 g of pectin was mixed with the orange juice. Satiety was measured on a visual analog scale before and at 0, 1, 2, 3 and 4 hours after orange juice and at 0, 30 and 60 minutes after ice cream. Multivariate ANOVA was used to examine satiety as a function of beverage (orange juice vs. orange juice plus pectin), time and pectin dose. RESULTS: There were significant differences in satiety as a function of beverage (p < 0.001) and time (p < 0.001) but not pectin dose (p = 0.121). The effect lasted up to 4 hours after ingesting pectin and orange juice and for 60 minutes after a second meal consisting of ice cream. CONCLUSIONS: Pectin in doses as small as 5 g mixed with orange juice increases satiety and can aid in a program to reduce weight by limiting food intake.

Adolescent↗