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. 2023 Jan 31:6:100451.
doi: 10.1016/j.crfs.2023.100451. eCollection 2023.

HPTLC-based fingerprinting: An alternative approach for fructooligosaccharides metabolism profiling

Luis Francisco Salomé-Abarca  1 , Ruth Esperanza Márquez-López  2 , Patricia Araceli Santiago-García  2 , Mercedes G López  1
Affiliations

HPTLC-based fingerprinting: An alternative approach for fructooligosaccharides metabolism profiling

Luis Francisco Salomé-Abarca et al. Curr Res Food Sci. .

Abstract

Fructans are categorized as fructose-based metabolites with no more than one glucose in their structure. Agave species possess a mixture of linear and ramified fructans with different degrees of polymerization. Among them, fructooligosaccharides are fructans with low degree of polymerization which might be approachable by high performance thin layer chromatography (HPTLC). Thus, this study used two emblematic Agave species collected at different ages as models to explore the feasibility of HPTLC-based fingerprinting to characterize fructooligosaccharides (FOS) production, accumulation, and behavior through time. To do so, high performance anion exchange was also used as analytical reference to determine the goodness and robustness of HPTLC data. The multivariate data analysis showed separation of samples dictated by species and age effects detected by both techniques. Moreover, linear correlations between the increase of the age in agave and their carbohydrate fraction was established in both species by both techniques. Oligosaccharides found to be correlated to species and age factors, these suggest changes in specific carbohydrate metabolism enzymes. Thus, HPTLC was proven as a complementary or stand-alone fingerprinting platform for fructooligosaccharides characterization in biological mixtures. However, the type of derivatizing reagent and the extraction color channel determined the goodness of the model used to scrutinize agavin fructooligosaccharides (aFOS).

Keywords: Agave; Data extraction; Fructooligosaccharides; HPTLC; Isomers.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
Unsupervised multivariate data analysis of fructooligosaccharide fractions of Agave angustifolia and Agave potatorum measured by high performance anion Exchange chromatography. A, Principal component analysis (PCA) scaled by Pareto and colored according to Agave species. B, PCA scaled by unit variance (UV) and colored according to Agave species. C, PCA scaled by Pareto and colored according to Agave age. D, PCA scaled by UV and colored according to Agave age. E, PCA of A. angustifolia specimens scaled by pareto and colored according to age. F, PCA of A. potatorum specimens scaled by pareto and colored according to age.
Fig. 2
Fig. 2
Supervised multivariate data analysis of fructooligosaccharide fractions of Agave angustifolia and Agave potatorum measured by high performance anion Exchange chromatography. A, Orthogonal projection to latent structures discriminant analysis (OPLSDA) scaled by pareto and classified according to Agave species. B, OPLS-DA scaled by unit variance (UV) and classified according to Agave species. C, OPLS-DA scaled by pareto and classified according to binary age; YT4, specimens younger than four years and OT3, specimens older than three years. D, Orthogonal projection to latent structures (OPLS) by UV including all Agave samples. E, (OPLS) by UV including only A. angustifolia samples. F, (OPLS) by UV including only A. potatorum samples.
Fig. 3
Fig. 3
HPTLC chromatograms of representative Agave angustifolia samples derivatized with A, aniline, B, α-naphthol, and C, orcinol. Sample number corresponds to sample age measured in years. STD corresponds to a standard compound mixture containing fructose/glucose, sucrose, 1-kestose, 1-nystose, and 1-F fructofuranosylnystose (DP-5) [2 mg/mL]. They appear from top to bottom in the mentioned order. RSE and RNE correspond to commercial samples of raftilose and raftiline, respectively.
Fig. 4
Fig. 4
Unsupervised multivariate data analysis of fructooligosaccharide fractions of Agave angustifolia and Agave potatorum measured by high performance thin layer chromatography. A, Principal component analysis (PCA) scaled by pareto and colored according to Agave species. B, PCA scaled by unit variance (UV) and colored according to Agave species. C, PCA scaled by pareto and colored according to Agave age. D, PCA scaled by UV and colored according to Agave age. E, PCA of A. angustifolia specimens scaled by pareto and colored according to age. F, PCA of A. potatorum specimens scaled by pareto and colored according to age.
Fig. 5
Fig. 5
Supervised multivariate data analysis of fructooligosaccharide fractions of Agave angustifolia and Agave potatorum measured by high performance thin layer chromatography. A, Orthogonal projection to latent structures discriminant analysis (OPLS-DA) scaled by pareto and classified according to Agave species. B, OPLS-DA scaled by pareto and classified according to binary age; YT4, specimens younger than four years and OT3, specimens older than three years. C, Orthogonal projection to latent structures (OPLS) by UV including all Agave samples. D, (OPLS) by UV including only A. angustifolia samples. E, (OPLS) by UV including only A. potatorum samples.
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