Functional Assessment of Prebiotics

The assessment of the potential health benefits of prebiotics can help in the discovery of new areas of action of prebiotics and the development of new products. Creative Enzymes' scientists utilize our advanced platform to employ the optimal analytical strategies, samples and instrumentation to maximize information recovery and provide accurate assessment results for our clients.

Introduction of Prebiotic Function Assessment

The health benefits of prebiotics are broad and include changes in microbiome composition and microbial metabolite levels, in addition to the multiple mechanisms that mediate health effects. Due to the complex potential impact of prebiotic metabolites on health, the choice of research methods is critical to further understanding of the field. In many researches of the effects of prebiotics, the focus has been on characterizing the composition of the gut microbiota using a range of microbiological techniques to assess the effects of prebiotics on the gut microbiota. In addition, the incorporation of prebiotics into different matrices such as food may affect the beneficial functions. Therefore, in addition to microbial composition analysis, functional assessment must be complemented by metabolite analysis. Creative Enzymes is committed to ensuring through our technologies that the actual health benefits conferred by prebiotics are measurable and certain.

Fig. 1 In vivo and in vitro evaluation of prebiotics and their components using different techniques. (Khangwal, 2019)

Our Services

There are many types of prebiotics, most of them are a subset of the carbohydrate group, mainly oligosaccharide carbohydrates, but also include other classes of compounds. Creative Enzymes can provide any prebiotic development and research services to our clients, including but not limited to the following.

• Digestion of Prebiotics

Creative Enzymes performs digestion tests of prebiotics by in vitro and in vivo methods. The main purpose is to imitate the human digestive tract in different stages of digestion. The in vitro simulation of gastric digestion of prebiotics is a complex process that is divided into three main tests：

• Gastric phase: enzymes, pepsin, and gastric lipase are given for 2 h at pH 2.5.
• After the gastric phase：digestion of prebiotics can be checked at the last stage, i.e., the small intestine phases.
• Small intestine phase: enzymes such as chymotrypsin, trypsin, colipase, and pancreatic lipase are given for 2 h at pH 6.

The in vivo digestion method was tested in animal models, mainly in rats and piglets. We assess the digestive properties of prebiotics by inserting a tube into the animal's gastrointestinal system or by measuring substrate digestion in feces.

• Fermentation Test

In the Creative Enzymes' laboratory, enteric flora fermentation prebiotic compounds are tested by in vitro and in vivo fermentation. In in vitro fermentation, we use a variety of methods to determine the fermentability of bacteria, including batch system, continues system, and the artificial gut system. The specific steps are to add the substrate at a known concentration to a vessel containing a fecal suspension or defined culture, incubate it under suitable conditions and sample it periodically. Our in vivo fermentation method is done using animal models. In the animal model, fecal samples are collected for assay after treatment with prebiotics.

• Analysis of Short-Chain Fatty Acids (SCF) and Gut Microorganisms

SCFs are produced from the fermentation of carbohydrates by gut microbiota. In addition, they directly increase the microbial populations. Creative Enzymes allows accurate analysis of compounds (e. g. short-chain fatty acids, branched-chain fatty acids, bile acids, indoles, cresols, ammonia, gases) present in biological samples due to host gut microbiota interactions.

We offer two main analytical platforms for metabolic analysis: 1H-NMR Spectroscopy and mass spectrometry (MS), often coupled with chromatographic techniques such as liquid chromatography (LC-MS) or gas chromatography (GC-MS), capable of simultaneously capturing hundreds or thousands of metabolites in a single sample. The acquired spectral data provide information about the presence, absence and concentration of metabolites.

In addition, our experimentalists measure changes in sample microbiota composition through a range of molecular biology techniques, including quantitative polymerase chain reaction, sequencing and analysis of 16S rRNA gene amplicons, DGGE (denaturing gradient gel electrophoresis), DNA microarray technology, and FISH (fluorescence in situ hybridization).

Service Process of Creative Enzymes

Creative Enzymes offers one-stop services for prebiotic production and evaluation services, providing our clients with time-saving and cost-effective solutions.

Creative Enzymes is a professional and experienced probiotic supplier and service provider. We are committed to evaluating and validating the function of prebiotics using a variety of in vitro and in vivo experiments to provide important insights into the development of the field. To get more information, please contact us and we will be happy to serve your research demands.

Reference

1. Khangwal, I.; et al. Prospecting prebiotics, innovative evaluation methods, and their health applications: a review. 3 Biotech. 2019, 9(5): 1-7.