Toxicology and Biodegradability of a Phthalate-Free and Bio-Based Novel Plasticizer.

Phthalate esters, mainly di-ethylhexylphthalate (DEHP), represent a class of chemicals primarily used as plasticizers for polyvinyl chloride in a wide range of domestic and industrial applications. These phthalate esters are low-toxicity environmental contaminants. To address these drawbacks, POLYSORB® ID 37, a blend of diesters obtained from esterification of isosorbide with plant-based fatty acids, was developed. The company can now offer PVC manufacturers a new product which competes with phthalates and other such chemicals. The market for plasticizers is very important, and ROQUETTE intends to provide a more sustainable and safer product. Isosorbide diester is bio-based (made from glucose and vegetable fatty acids). This plasticizer is registered in REACH regulation for high volumes (>1000 T/year). Risk assessment was obtained by conducting a wide range of biodegradability and toxicological protocols, using rodent models, according to established guidelines. Overall, all of the toxicological and biodegradability studies demonstrated that POLYSORB® ID 37 is nontoxic to mammalian life and is readily biodegradable.

Evaluation of soluble corn fiber on chemical composition and nitrogen-corrected true metabolizable energy and its effects on in vitro fermentation and in vivo responses in dogs.

Dietary fermentable fiber is known to benefit intestinal health of companion animals. Soluble corn fiber (SCF) was evaluated for its chemical composition, nitrogen-corrected true ME (TMEn) content, in vitro digestion and fermentation characteristics, and in vivo effects on nutrient digestibility, fecal fermentation end products, and modulation of the fecal microbiome of dogs. Soluble corn fiber contained 78% total dietary fiber, all present as soluble dietary fiber; 56% was low molecular weight soluble fiber (did not precipitate in 95% ethanol). The SCF also contained 26% starch and 8% resistant starch and had a TMEn value of 2.6 kcal/g. Soluble corn fiber was first subjected to in vitro hydrolytic-enzymatic digestion to determine extent of digestibility and then fermented using dog fecal inoculum, with fermentative outcomes measured at 0, 3, 6, 9, and 12 h. Hydrolytic-enzymatic digestion of SCF was only 7%. In vitro fermentation showed increased (P < 0.05) concentrations of short-chain fatty acids through 12 h, with acetate, propionate, and butyrate reaching peak concentrations of 1,803, 926, and 112 µmol/g DM, respectively. Fermentability of SCF was higher (P < 0.05) than for cellulose but lower (P < 0.05) than for pectin. In the in vivo experiment, 10 female dogs (6.4 ± 0.2 yr and 22 ± 2.1 kg) received 5 diets with graded concentrations of SCF (0, 0.5, 0.75, 1.0, or 1.25% [as-is basis]) replacing cellulose in a replicated 5 × 5 Latin square design. Dogs were first acclimated to the experimental diets for 10 d followed by 4 d of total fecal collection. Fresh fecal samples were collected to measure fecal pH and fermentation end products and permit a microbiome analysis. For microbiome analysis, extraction of DNA was followed by amplification of the V4 to V6 variable region of the 16S rRNA gene using barcoded primers. Sequences were classified into taxonomic levels using a nucleotide basic local alignment search tool (BLASTn) against a curated GreenGenes database. Few changes in nutrient digestibility or fecal fermentation end products or stool consistency were observed, and no appreciable modulation of the fecal microbiome occurred. In conclusion, SCF was fermentable in vitro, but higher dietary concentrations may be necessary to elicit potential in vivo responses.

In vivo efficacy of microbiota-sensitive coatings for colon targeting: a promising tool for IBD therapy.

The first proof of concept in vivo for a new type of microbiota-sensitive film coatings allowing for colon targeting is presented. The efficacy of these polysaccharide barriers to optimize drug release for the treatment of inflammation is demonstrated in an experimental colitis model with Wister rats. 5-Aminosalicylic acid (5-ASA) pellets were prepared by extrusion-spheronization and coated with Nutriose:ethylcellulose (EC) 1:4 or peas starch:ethylcellulose 1:2 blends. The pellets were mixed with standard chow, and the daily drug dose was 150mg/kg. For reasons of comparison, also commercially available Pentasa pellets and placebo pellets were studied. At day 3 after the beginning of the treatment, colitis was induced by intrarectal administration of trinitrobenzene sulfonic acid (TNBS). Animals were sacrificed on day 6. Macroscopic and histological evaluations of colitis were performed blindly. In addition, inflammatory markers were evaluated using ELISA and real-time PCR. Rats receiving TNBS and placebo pellets developed a severe colitis in the distal half of the colon. 5-ASA administered in the form of Pentasa pellets reduced macroscopic inflammation by only 5%. In contrast, the colon lesions were much less severe upon treatment with Nutriose:EC- and peas starch:EC-coated pellets: The macroscopic score was reduced by 25 and 24%, respectively. Decreases of 37 and 38% of the histological lesions confirmed the efficacy of these new colon targeting systems. Also, inflammatory markers (MPO, IL-1ß mRNA, TNF mRNA) were significantly decreased in rats receiving Nutriose:EC- and peas starch:EC-coated pellets compared to Pentasa pellets. Furthermore, real-time PCR analysis indicated increased activation of the target receptor PPAR-γ and the HMGCS2 gene in rats upon administration of 5-ASA loaded Nutriose:EC- and peas starch:EC pellets compared to the commercial product. Also, HPLC-MS/MS analysis of plasma samples demonstrated that the level of the main metabolite of the drug (N-acetyl-5-ASA) was much lower upon administration of Nutriose:EC or peas starch:EC coated pellets compared to Pentasa pellets, indicating that undesired premature drug release in the upper gastrointestinal tract was more effectively hindered. In addition to the rat study, in vivo imaging of transgenic mice expressing the luciferase gene evidenced much more pronounced PPAR-γ activation upon 5-ASA administration in the form of Nutriose:EC-coated pellets versus Pentasa pellets. All these results clearly demonstrate the superiority of these microbiota-sensitive polysaccharide-based film coatings for colon targeting in vivo.

Effects of maltitol and xylitol chewing-gums on parameters involved in dental caries development.

AIM: The effects on plaque parameters of sugar free chewing-gums (CG) sweetened with either maltitol or xylitol were assessed to better understand the role polyols can play in dental caries prevention. MATERIALS AND METHODS: A double-blind, parallel, randomised, controlled study was conducted in China. Subjects (N = 258, age = 13 to 15 years-old) were divided into 4 groups: 2 receiving polyols CG, containing respectively maltitol or xylitol, a group receiving gum base (placebo) and a negative control group not receiving any gum. CG were chewed for 30 days. This corresponds to a 10 g consumption of polyol per day. Plaque parameters (growth, pH, bacteria and insoluble glucans) were evaluated throughout the experimental period. RESULTS: All parameters studied were significantly modified with gum base compared to no-gum: plaque pH increased; plaque growth, bacteria (S. mutans, S. sobrinus, A. viscosus and Lactobacillus) and insoluble glucans decreased. Maltitol and xylitol CG led similarly to a higher plaque pH (AUC, p⋜0.05) on short (at baseline after the first CG consumption) and long term (after 4 weeks of daily CG consumption), with or without saliva stimulation compared to both control and placebo groups. They led to a decrease in plaque growth (p=0.02) over the experimental period compared to controls. Moreover, they significantly reduced the concentration of 4 cariogenic bacteria species (p⋜0.05) in dental plaque compared to gum base. CONCLUSION: Sugar free CG sweetened with either maltitol or xylitol can similarly reduce plaque acidogenicity compared to gum base through a decrease in oral bacteria presence. The use of a gum base placebo allowed to isolate effects on parameters involved in dental caries development specific to maltitol and xylitol, and to show these effects were similar.

Potato fiber as a dietary fiber source in dog foods.

Potato fiber (PF), a coproduct of potato starch manufacture, was evaluated as a potential novel fiber source in dog food. Potato fiber contained 55% total dietary fiber, 29% starch, 4% crude protein, and 2% acid-hydrolyzed fat. The PF substrate was evaluated for chemical composition, in vitro digestion and fermentation characteristics, and in vivo responses. For the in vitro hydrolytic-enzymatic digestion and fermentation experiment, raw and cooked PF substrates were first subjected to hydrolytic-enzymatic digestion to determine OM disappearance and then fermented using dog fecal inoculum. Fermentation characteristics were then measured at 0, 3, 6, 9, and 12 h. For the in vivo experiment, 10 female mixed-breed dogs (6.13±0.17 yr; 22±2.1 kg) were provided 5 diets with graded concentrations (0%, 1.5%, 3%, 4.5%, or 6%) of PF in a replicated 5×5 Latin square design. Dogs were acclimated to the test diet for 10 d, followed by 4 d of total fecal collection. Fresh fecal samples were collected to measure fecal pH and fermentation end products. In vitro digestion revealed that raw and cooked PF were 32.3% and 27.9% digested enzymatically, whereas in vitro fermentation showed that PF was fermentable through 9 h. Raw PF had greater (P<0.05) acetate, propionate, and total short-chain fatty acid (SCFA) concentrations at the 12-h time point compared with cooked PF. The in vivo experiment showed no differences in apparent total tract DM, OM, CP, acid-hydrolyzed fat, or energy digestibility of diets containing graded concentrations of PF. However, total dietary fiber digestibility exhibited a linear increase (P<0.01) with increasing PF concentrations in the diet. Overall, linear increases (P<0.01) were observed for all individual and total SCFA, with a concomitant linear decrease (P<0.01) in fecal pH with increasing dietary PF. Fecal protein catabolite concentrations were low or undetectable, with the exception of spermidine, which exhibited a linear increase with increasing concentrations of PF. These findings indicated that inclusion of PF elicited favorable fermentation characteristics without negatively affecting nutrient digestibility or stool characteristics, indicating that PF could be a functional dietary fiber source in dog foods.

Impact of a resistant dextrin on intestinal ecology: how altering the digestive ecosystem with NUTRIOSE®, a soluble fibre with prebiotic properties, may be beneficial for health.

OBJECTIVES: The prebiotic potential of NUTRIOSE®--a sugar-free, digestion-resistant dextrin--was evaluated in two randomized, placebo-controlled trials that included 48 and 40 healthy volunteers, respectively. METHODS: In study 1, the effect on colonic bacteria of NUTRIOSE® 10, 15 or 20 g/day administered for 14 days was examined; in study 2, gut microbial changes in response to NUTRIOSE® 8 g/day for 14 days were monitored using real-time polymerase chain reaction analysis. RESULTS: NUTRIOSE® increased proliferation of Bacteroides and inhibited Clostridum perfringens in both studies, increased ß-glucosidase activity (at 10 and 15 g/day) and decreased colonic pH (at 20 g/day). The increase in short-chain fatty acid production with NUTRIOSE® consumption was not statistically significant. There were no indications of gastrointestinal intolerance at any dose. CONCLUSIONS: According to commonly accepted definitions, NUTRIOSE® is a prebiotic soluble fibre that provides a beneficial effect on colonic ecology while preserving digestive comfort.

Non-coated multiparticulate matrix systems for colon targeting.

BACKGROUND: Colon specific drug delivery can significantly improve the efficacy of local treatments of inflammatory bowel diseases. Film coatings containing the starch derivative Nutriose have recently been reported to minimize 5-ASA release in media simulating the upper gastro intestinal tract (GIT), while releasing the drug in a time-controlled manner upon contact with feces from Crohn's Disease and Ulcerative Colitis patients. It was the aim of this study to prepare Nutriose-containing matrix pellets and mini tablets in order to avoid a film coating step. METHODS: Highly dosed matrix pellets were prepared by extrusion-spheronization, highly dosed mini tablets by compression. Various types of lipids were added and drug release measured in 0.1 N HCl and phosphate buffer pH 6.8, optionally containing pepsin and pancreatin. RESULTS: The type of added lipid and the preparation technique, in particular the curing conditions, significantly affected the resulting drug release kinetics. Glyceryl palmitostearate containing pellets and mini tablets showed the most promising results upon appropriate curing, minimizing premature drug release in media simulating the upper GIT. CONCLUSION: The proposed novel multiparticulates do not require a film coating step and show an interesting potential for site-specific drug delivery to the colon of inflammatory bowel disease patients.

Safety profile of a food dextrin: acute oral, 90-day rat feeding and mutagenicity studies.

Nutriose is a glucose polysaccharide produced by the chromatographic separation of a dextrin fraction derived from maize, wheat or other edible starches. Animal safety studies conducted on Nutriose FB are reported. They include an acute oral and a 90-day study in rats and short-term in bacteria (Ames test) and a mutation assay at the TK locus in L5178Y mouse lymphoma cells. An acute oral study in Sprague-Dawley rats established the LD(50) as greater than 2000 mg/kg. In a 90-day, oral subchronic study, Sprague-Dawley rats were administered Nutriose FB in their diet at doses of 0, 1.25%, 2.5% or 5% for 13 weeks. Neither mortality nor significant behavioral changes occurred during the study. The consumption of Nutriose FB did not have any effect on body weight or on feed or water consumption. Blood coagulation and hematology and blood and urine biochemistry did not reveal any toxic effect of the compound. No treatment-related histopathological differences were observed between control and test groups. Adverse clinical observations, including ophthalmological observations, were marginal and not considered treatment-related. There was no effect of Nutriose FB on relative or absolute organ weight of rats of either sex, except for the increase in caecum content and caecum mucosa. The increase in caecum weight is considered a physiological adaptation seen after the ingestion of indigestible carbohydrates and is not considered a toxicological effect. The No-Observed-Adverse-Effect-Levels (NOAELs) were established by the highest tested doses: 4.4 g/kg bw/day in males and 6.5 g/kg bw/day in females. Mutation assays in bacteria (Ames tests) and in mammalian cells (tk locus in mouse lymphoma cells) were negative with Nutriose FB.

Long-term gastrointestinal tolerance of NUTRIOSE FB in healthy men.

OBJECTIVE: To determine the gastrointestinal (GI) tolerance of NUTRIOSE FB in men. DESIGN: A randomized, placebo-controlled, parallel, double-blind study. SETTING: The metabolic ward of TNO Quality of Life. SUBJECTS: Forty-eight subjects started the study: 16 men participated in one of the three treatments. SUBJECTS consumed either 22.5 g of pure maltodextrin (Glucidex 6), or 30 or 45 g of the dextrin NUTRIOSE FB daily for 4-5 weeks. Forty-three subjects completed the study (age: 34.7 +/- 8.2 years; BMI 24.9 +/- 3.3 kg m2). MEASUREMENTS: Tolerance of NUTRIOSE FB was examined with a GI complaints questionnaire; effectiveness on colonic flora was examined by faecal analysis; fermentation by breath hydrogen excretion measurement. Furthermore, the effect on body weight (BW), energy intake and blood parameters were examined in the study. RESULTS: Both doses of NUTRIOSE FB were very well tolerated and GI complaints hardly differed from the placebo treatment. No diarrhoea was reported due to NUTRIOSE FB supplementation. In the course of the study, some habituation and adaptation of GI symptoms were found. Fermentation and faecal characteristics (pH and enzyme activity) were significantly positively affected with NUTRIOSE FB treatment. Body weight in both NUTRIOSE FB groups remained stable over time, although the placebo-treated group showed a small increase in BW (Deltaday35-1 0.8 +/- 1.0 kg) (P = 0.07). However, total food intake and macronutrient composition of the diet remained the same throughout the study. No significant differences were found between the three treatment groups in hunger and satiety scores and food preferences. CONCLUSIONS: Long-term supplementation of 30 or 45 g of the dextrin NUTRIOSE FB per day was well tolerated, and may act as a pre-biotic supplement. SPONSORSHIP: TNO Quality of Life was assigned by Roquette Frères to perform the study.

Energy value of a low-digestible carbohydrate, NUTRIOSE FB, and its impact on magnesium, calcium and zinc apparent absorption and retention in healthy young men.

BACKGROUND: Long-term consumption of imbalanced diets, poor in dietary fibres, resulted in the prevalence of several nutritional pathologies. However, low digestible carbohydrates (LDC) have many beneficial effects, especially on energy intake, digestive physiology, and mineral absorption. AIM OF THE STUDY: To determine the digestive effects of a LDC, called NUTRIOSE FB, its metabolisable energy (ME) value, and its effects on mineral absorption in humans. METHODS: Ten healthy young men were fed for 31 d periods a maintenance diet supplemented with either dextrose or the LDC at a level of 100 g DM/d, in six equal doses per d according to a cross-over design. After a 20 d adaptation period, food intake was determined for 11 days using the duplicate meal method, and faeces and urine were collected for 10 d for further analyses. RESULTS: Ingestion of the LDC did not cause severe digestive disorders, except excessive gas emission, and flatulence and slight abdominal pain in some subjects for intakes above 50 g DM/d. Wet and dry stool outputs increased by 45 and 70%, respectively (P<0.02). In vitro enzymatic digestibility of the LDC was 15 (SD 1.5) %, and 9.2 (SD 8.3) % of the LDC was excreted in faeces (P<0.001). The ME value of the LDC was 14.1 (SD 2.3) kJ/g DM, that is 14 % less than the tabulated values of sucrose and starch. Its net energy value (NEV), estimated using three prediction equations, was 8.7, 8.9, and 11.4 kJ/g DM. Ingestion of the LDC significantly increased the relative apparent absorption of Mg, and Mg retention by 67% and 31 mg/d, respectively, tended to increase Ca apparent absorption (P=0.110) and Ca retention (P=0.059), but did not significantly alter Zn parameters. CONCLUSION: NUTRIOSE FB can be used as a "bulking" agent, and substituted up to 50 g/d for usual maltodextrins without causing digestive disorders in healthy subjects. It would reduce intestinal transit disorders and energy intake, and improve magnesium and calcium absorption and retention.

Short-term digestive tolerance of different doses of NUTRIOSE FB, a food dextrin, in adult men.

OBJECTIVE: To determine the tolerance of increasing dosages of an incompletely hydrolysed and/or incompletely absorbed food dextrin coming from wheat starch, NUTRIOSE FB, at daily levels of 10 and 15 g up to 60 and 80 g, respectively. DESIGN: A randomized, double-blind, multiple dose, placebo-controlled, combined crossover and parallel trial. SETTING: The metabolic ward of TNO Nutrition and Food Research. SUBJECTS: A total of 20 healthy men (age 31.7 +/- 9.1 y; BMI 24.5 +/- 2.9 kg/m2). INTERVENTION: One group of 10 subjects consumed on top of their diet 10, 30 and 60 g of NUTRIOSE FB or maltodextrin (placebo) daily. The other group of 10 subjects consumed 15, 45 and 80 g daily. Each dose was consumed for 7 days. RESULTS: Compared with placebo, flatulence occurred more frequently over the last 6 days on 30, 60 or 80 g/day of NUTRIOSE FB (P < 0.05). During the last 24 h, that is, days 6-7, of 60 and 80 g/day of NUTRIOSE FB, the frequency of flatulence was even higher (P < 0.05). During the last 24 h on a daily dose of 60 g NUTRIOSE FB, the frequency of defecation decreased (P < 0.05). Bloating occurred more often during the last 24 h on 80 g/day of NUTRIOSE FB (P < 0.05). None of the doses of NUTRIOSE FB resulted in diarrhoea. Compared to baseline levels, breath H2 excretion, which was only measured after a week with 10 and 15 g of NUTRIOSE FB daily, increased (P < 0.05). However, no difference in area under the curve was found. CONCLUSIONS: NUTRIOSE FB is a fermentable carbohydrate and is well tolerated up to a dose of 45 g daily. Higher daily dosages (60 and 80 g) may result in flatulence, but does not result in diarrhoea. SPONSORSHIP: TNO Nutrition and Food Research was assigned by Roquette Frères to perform the study.

Glycaemic and insulinaemic responses to a new hydrogenated starch hydrolysate in healthy and type 2 diabetic subjects.

BACKGROUND: Industrialists are searching for a sugar replacement in confectioneries such as hard candies, gum and chocolate. Lycasin HBC is a suitable candidate. Nevertheless, no information on its plasma glucose and insulin responses exists. Therefore, we aimed to evaluate the glycaemic and insulinaemic indices of Lycasin HBC in healthy subjects and in subjects with type 2 diabetes mellitus. METHODS: Six healthy and six type 2 diabetic men participated in the study. Each subject absorbed, after an overnight fast, a challenge of either 50 g of glucose or 50 g of Lycasin HBC using a randomised double-blind crossover design. Blood samples for measuring plasma glucose and insulin concentrations were collected during a 3 hour period. RESULTS: The calculated glycaemic index of Lycasin HBC was 47 +/- 10% in healthy subjects and 25 +/- 6% in patients with type 2 diabetes mellitus. The insulinaemic index of Lycasin HBC was 23 +/- 4% and 39 +/- 14%, respectively. As glucose levels oscillate in a very limited range in normal healthy subjects, the insulinaemic index must be considered here. On the other hand, it is the glycaemic rather than the insulinaemic index that must be assessed in diabetic subjects due to impairment of insulin secretion. CONCLUSIONS: The tested Lycasin HBC showed a low insulinaemic index in healthy subjects (23 +/- 4%) and a low glycaemic index (25 +/- 6%) in type 2 diabetic patients. Thus, it might be considered as an interesting sucrose substitute in confectionery for individuals with or without diabetes.

The comparative gastrointestinal responses of children and adults following consumption of sweets formulated with sucrose, isomalt and lycasin HBC.

OBJECTIVES: To determine the gastrointestinal responses of children and adults following consumption of sucrose, isomalt and lycasin HBC and to compare these at two different dose levels in adults. DESIGN: Both studies were randomised, double-blind, cross-over designs. SUBJECTS: Fifty-one children aged 6-9 y were recruited from primary schools in the Salford area of Greater Manchester. Forty-eight children completed the study. Fifty healthy adult volunteers aged 18-24 y were recruited from the student population of the University of Salford. All subjects completed the study. INTERVENTIONS: Children consumed either 25 g of sucrose, isomalt or lycasin HBC and adults 25 and 40 g in hard boiled sweets per day for two consecutive test days. Test periods of 2 days were separated by 7 day washout periods. Children consumed sweets throughout test days and adults in no less than 30 min but no more than 90 min. Subjects reported the prevalence and magnitude of flatulence, borborygmi, bloating, colic, bowel movements and watery faeces. RESULTS: Consumption of 25 g isomalt provoked a mild laxative effect in children but not in adults. Consumption of 25 g isomalt significantly increased the prevalence and magnitude of gastrointestinal responses in both children and adults. Consumption of 25 g lycasin HBC significantly increased borborygml in children and adults but no other gastrointestinal responses. Consumption of 40 g lycasin HBC or isomalt by adults significantly increased the mean frequency of bowel movements and the number of subjects passing watery faeces. In adults, 40 g isomalt and lycasin HBC provoked significantly more gastrointestinal responses compared to 25 g of either product. CONCLUSIONS: Consumption of 25 g lycasin HBC does not provoke an unacceptable laxative effect or gastrointestinal response in children or adults compared to 25 g isomalt, which is associated with a mild laxative effect and increase in gastrointestinal responses. In adults gastrointestinal responses following consumption of products were found to be dose dependent.

Net energy value of two low-digestible carbohydrates, Lycasin HBC and the hydrogenated polysaccharide fraction of Lycasin HBC in healthy human subjects and their impact on nutrient digestive utilization.

The metabolizable energy content of low-digestible carbohydrates does not correspond with their true energy value. The aim of the present study was to determine the tolerance and effects of two polyols on digestion and energy expenditure in healthy men, as well as their digestible, metabolizable and net energy values. Nine healthy men were fed for 32 d periods a maintenance diet supplemented either with dextrose, Lycasin HBC (Roquette Frères, Lestrem, France), or the hydrogenated polysaccharide fraction of Lycasin HBC, at a level of 100 g DM/d in six equal doses per d according to a 3 x 3 Latin square design with three repetitions. After a 20 d progressive adaptation period, food intake was determined for 12d using the duplicate meal method and faeces and urine were collected for 10 d for further analyses. Subjects spent 36 h in one of two open-circuit whole-body calorimeters with measurements during the last 24h. Ingestion of the polyols did not cause severe digestive disorders, except excessive gas emission, and flatulence and gurgling in some subjects. The polyols induced significant increases in wet (+45 and +66% respectively, P<0.01) and dry (+53 and +75 % respectively, P<0.002) stool weight, resulting in a 2% decrease in dietary energy digestibility (P<0.001). They resulted also in significant increases in sleeping (+4.1%, P<0.03) and daily energy expenditure (+2.7 and +2.9% respectively, P<0.02) compared with dextrose ingestion. The apparent energy digestibility of the two polyols was 0.82 and 0.79 respectively, their metabolizable energy value averaged 14.1 kJ/g DM, and their net energy value averaged 10.8 kJ/g DM, that is, 35 % less than those of sucrose and starch.