Thin-layer Chromatography and Matrix-assisted Laser Desorption/Ionization Mass Spectrometric Analysis of Oligosaccharides in Biological Samples
| Authors | |
|---|---|
| Year of publication | 2007 |
| Type | Article in Periodical |
| Magazine / Source | JPC-J. Planar Chromatogr.-Mod. TLC |
| MU Faculty or unit | |
| Citation | |
| Field | Analytic chemistry |
| Keywords | TLC; MALDI MS; oligosaccharides |
| Description | TLC and MALDI MS of Oligosaccharides in Biological Samples. Thin-layer chromatography (TLC) and matrix-assisted laser desorption/ ionization mass spectrometry (MALDI MS) have been used for analysis of fructooligosaccharides (FOS), used as feed additives (prebiotics), in biological samples. The contents of different regions of a piglet’s intestine (jejunum, ileum, cecum, and colon) were analyzed. Raftilose, a commercially available dietetic product which contains fructooligosaccharides, was added into the feed of the weaning piglets. Complicated biological samples for TLC analysis were used with minimal pretreatment. Analysis was performed on glass-backed precoated silica gel layers impregnated with sodium acetate and developed with butanol–acetic acid–water, 2 + 2 + 1 (v/v), in a vertical glass twin-trough chamber with mobile phase vapor saturation. Visualization of FOS on chromatograms was achieved with a mixture of diphenylamine, aniline, and phosphoric acid in acetone as primary detection reagent. Blue–pink spots of FOS on chromatograms were also detected by reflectance densitometry at lambda = 370 nm. Optimum MALDI MS conditions for analysis of Raftilose standard were determined. By use of 10 mg mL–1 2,5-dihydroxybenzoic acid solution and addition of 1 mg mL–1 sodium acetate, a limit of detection (LOD) of 1.4 x 10–4% Raftilose was achieved; this corresponded to ~0.5 ng Raftilose deposited on the target. The effect of biological medium (intestine contents) on the MALDI spectra of Raftilose was checked. LODs of Raftilose in the presence of biological medium were determined from the spectra of samples taken from the ileum and jejunum spiked with 0.05 mg mL–1 Raftilose standard. |
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