Abstract
Sialyllactose (SL), an acidic oligosaccharide, has immune-protective effects against pathogens and helps with the development of the immune system and intestinal microorganisms. To elucidate the pharmacokinetic characterization after oral administration to rats, the simultaneous quantification method for 3′-SL and 6′-SL in rat plasma was validated, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in an electrospray ionization (ESI) mode. Several types of columns [C18, amide, and hydrophilic interaction liquid chromatography (HILIC) phase] were used to separate the peaks of 3′-SL and 6′-SL, which improved chromatographic selectivity. Ultimately, the HILIC phase column had a good peak shape and quick resolution, with a mobile phase comprising ammonium acetate buffer and acetonitrile obtained by gradient elution. In addition, the simultaneous quantification of 3′-SL and 6′-SL in rat plasma samples were adequately applied to pharmacokinetic study.
Highlights
Sialyllactose (SL), a kind of sialyloligosaccharide, is an oligosaccharide with sialic acid and a lactose group [1]
We found endogenous sialyllactose in rat plasma, and a calibration curve spiked standard solution in a blank matrix cannot be used, since 3 - and 6 -sialyllactose (6 -SL) are endogenous compounds
Considering the sensitivity, resolution, asymmetry, and the buffer capacity of ammonium acetate ranging from pH 3.8 to 5.8, the mobile phase was set to 10 mM ammonium acetate buffer and the acetonitrile (Figure 1B)
Summary
Sialyllactose (SL), a kind of sialyloligosaccharide, is an oligosaccharide with sialic acid and a lactose group [1]. The most abundant sialyloligosaccharides in human milk are 3 sialyllactose (3 -SL), 6 -sialyllactose (6 -SL), sialyllacto-N-tetraose (SLNT), and disialyllactoseN-tetraose (DSLNT) [2,3]. Human milk oligosaccharides (HMOs) have low sensitivity to UV detection and are not well retained in reversed-phase analytical columns, such as the C18 column. Most of the conventional methods measured HMO levels in milk, and only Santos-Fandila’s method analyzed sialyllactose levels in plasma or serum. HMOs in serum were extracted into chloroform/methanol (2:1, v/v) and were precipitated by ethanol, overnight, at 4 ◦C. The time for sample preparation was reduced, as compared to that of the overnight method
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