Dietary Sugars Analysis: Quantification of Fructooligossacharides during Fermentation by HPLC-RI Method

Abstract

In this work, a simple chromatographic method is proposed and in-house validated for the quantification of total and individual fructooligossacharides (e.g., 1-kestose, nystose, and 1F-fructofuranosylnystose). It was shown that a high-performance liquid chromatography with refractive index detector could be used to monitor the dynamic of fructooligossacharides production via sucrose fermentation using Aspergillus aculeatus. This analytical technique may be easily implemented at laboratorial or industrial scale for fructooligossacharides mass-production monitoring allowing also controlling the main substrate (sucrose) and the secondary by-products (glucose and fructose). The proposed chromatographic method had a satisfactory intra- and inter-day variability (in general, with a relative standard deviation lower than 5%), high sensitivity for each sugar (usually, with a relative error lower than 5%), and low detection (lower than 0.06 ± 0.04 g/L) and quantification (lower than 0.2 ± 0.1 g/L) limits. The correct quantification of fructooligossacharides in fermentative media may allow a more precise nutritional formulation of new functional foods, since it is reported that different fructooligossacharides exhibit different biological activities and effects.

Towards smart biomanufacturing: a perspective on recent developments in industrial measurement and monitoring technologies for bio-based production processes

“…Furthermore, combined analytical and chemometric approaches are being developed to monitor the substrate, metabolites and products in bioprocesses. Chemometrics are employed to provide real-time analysis of variables that otherwise would require off-line analysis and interpretation using methods such as HPLC or GC/MS [19]. A few examples include in-situ biosensors [20][21][22], optical sensors [23,24], at-line implementations of traditional methods (HPLC, GC/MS) [19,25] and spectroscopic sensors [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42], among others.…”

Development and Optimisation of HILIC-LC-MS Method for Determination of Carbohydrates in Fermentation Samples

“…The mass spectrometric detection helps with the selectivity of the methodology as mass spectrometer could filter out matrix interfering components thus providing cleaner and unambiguous spectra. The simultaneous measurement of fructose, glucose, galactose, sucrose, and lactose could be done in a fraction of time and less consumed solvent compared to classical methods used elsewhere [ 10 , 11 , 16 , 32 ]. The method developed here suits for both to identify and characterise the metabolism of various starter cultures (like here with Streptococcus thermophilus ) as well as to detect the sugar profile of different food matrices.…”

“…The main path to analyse carbohydrates was to use either gas chromatography with derivatisation of saccharides or liquid chromatography employing ion-exchange resins [ 8 ]. The development of novel stationary phases for both gas and liquid chromatography increased the number of applications where carbohydrate analysis could be performed from samples obtained from various sources, i.e., raw nutritional materials, animals, bacteria, humans, and so forth [ 9 , 10 , 11 , 12 ]. At the same time, developments in ion chromatography and electrophoresis allowed to measure carbohydrates in similar matrices creating the alternative ways for the measurements of saccharides [ 13 , 14 , 15 , 16 ].…”

Liquid Chromatography Analysis of Common Nutritional Components, in Feed and Food

“…Mild conditions used during hydrolysis avoid the release from other naturally occurring saccharides in wheat that would otherwise interfere during the fructan estimation (e.g., raffinose oligosaccharides) [316]. Correia and coworkers have developed a method to analyze fructooligosaccharides [317]. These sugars are dietary and are used as food ingredients (incorporated as dietary fibers in commodities).…”