Uptake and metabolism of pyridoxine-4′-α-D-glucoside and pyridoxine-5′-α-D-glucoside by freshly isolated rat liver cells were studied using the membrane filtration method. The presence of the pyridoxine-α-glucosides at either 0.5 or 5.0 µmol/L had no inhibitory effect on the uptake of pyridoxine (0.5 µmol/L) by hepatocytes. The amount of pyridoxine-α-glucoside transported into the cells was only 10–12% of that of pyridoxine. The initial rates of uptake of the pyridoxine-α-glucosides were linear with concentrations up to 50 µmol/L. On entry into the cells, 78% of pyridoxine-5′-α-glucoside and 15% of the pyridoxine-4′-α-glucoside were hydrolyzed at the end of 60 min. Because pyridoxine-5′-α-glucoside is hydrolyzed readily and does not interfere with the transport of pyridoxine, the glucoside, if present in animal- or plant-derived foods, may be available as a source of vitamin B-6.