Abstract
This work presents a comparative study for the analysis of carbohydrates for four common chromatographic methods, each coupled to mass spectrometry. Supercritical fluid chromatography (SFC), hydrophilic interaction liquid chromatography (HILIC), reversed-phase liquid chromatography (RP-LC) and gas chromatography (GC) with detection by triple quadrupole mass spectrometer (QqQ-MS) are compared. It is shown that gas chromatography and reversed-phase liquid chromatography, each after derivatisation, are superior to the other two methods in terms of separation performance. Furthermore, comparing the different working modes of the mass spectrometer, it can be determined that a targeted analysis, i.e. moving from full scan to single ion monitoring (SIM) and multiple reaction monitoring (MRM), results in an improvement in the sensitivity as well as the repeatability of the method, which has deficiencies especially in the analysis using HILIC. Overall, RP-LC–MS in MRM after derivatisation with 1-phenyl-3-methyl-5-pyrazolone (PMP) proved to be the most suitable method in terms of separation performance, sensitivity and repeatability for the analysis of monosaccharides. Detection limits in the nanomolar range were achieved, which corresponds to a mass concentration in the low µg/L range. The applicability of this method to different biological samples was investigated with various herbal liquors, pectins and a human glycoprotein.Graphical abstract
Highlights
Carbohydrates, called saccharides or sugars, represent one of the most important biochemical substance classes and have a central role in the metabolism of every living organism
Carbohydrate intake further contributes to diseases and excessive or regular consumption has been associated with diabetes, dental issues and attention deficit hyperactivity disorder (ADHD) among others [2,3,4]
Carbohydrates can appear as monosaccharides or more commonly as oligo- and polysaccharides, but to evaluate the native sequence of the saccharide chains, the evaluation of the monosaccharide moieties is necessary
Summary
Carbohydrates, called saccharides or sugars, represent one of the most important biochemical substance classes and have a central role in the metabolism of every living organism. Carbohydrates, in the form of naturally occurring or added sugars, constitute the most important dietary energy source for humans, and total sugars are consumed in varying quantities depending on the age group, between 13% for adults and 38% for infants in terms of total energy intake [1]. The analysis of sugar monomers in samples can be carried out using various chromatographic methods. Liquid chromatographic (LC) methods are used to take advantage of the various available stationary phases. Separation of sugars has been carried out using amino and amide as well as HILIC phases, which separate the carbohydrates in their
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