9Foods contain the family of vitamin B 6 compounds which consists of six forms, i.e., pyridoxine (PN), pyridoxal (PL), pyridoxamine (PM), pyridoxine 5′-phosphate (PNP), pyridoxal 5′-phosphate (PLP), and pyridoxamine 5′-phosphate (PMP) (Fig. 1). PLP is a coenzyme for many enzymes involved in amino acid and carbohydrate metabolism, and plays a key role in the nutritional function of vitamin B 6. The other forms show the same nutritional efficiency because they are converted into PLP in cells. Plants contain a storage form of vitamin B6, pyridoxine-b-glucoside (PNG) (1), which is not counted as a vitamin B6 form although its utilization is 58% relative to PN (2). 4-Pyridoxic acid (4-PA), which exhibits no vitamin B6 activity (3), was found in some foods, such as milk.The composition of vitamin B6 and the related compounds in human milk has been extensively studied, because breast milk is the sole source of nutrients for almost all infants up to 4-6 mo. The total content of vitamin B6 and the distribution of the vitamin B6 forms in the milk of American women have been investigated by high performance liquid chromatography (HPLC) in several laboratories (1,(4)(5)(6)(7). The studies showed that the mature milk samples contained approximately 1 mmol/L of total vitamin B6, in which PL was the major (65.9-82.8% of the total content) form in the milk. PLP was the second (12.5-31%) in the mature milk although it was not found in the transitional milk (7). The contents of the other forms were very low: PM (0-5.2% and 11.1% in the mature and transitional milk, respectively), PMP (0-1.9% and 0%), and PN (0-4.4% and 11.8%). So far, PNP has not been found. 4-PA content was approximately 0.2 (mmol/L) (8).In contrast to the detailed studies on the milk of American women, only one study has reported on the total content of vitamin B6 and the distribution of vitamin B6 forms in milk of Japanese women. Average total content of vitamin B6 in 25 mature milk samples determined by HPLC was approximately 1.45 (mmol/L) (9). The contents of PLP and PL were 1.15 and 0.30 (mmol/L), respectively; the relative content of PLP and PL was the reverse of that found in the American wom- Summary The contents of six vitamin B6 forms, pyridoxine-b-glucoside, and 4-pyridoxic acid in mature milk of 20 Japanese lactating women consuming ordinary Japanese foods were determined by a 4-pyridoxolactone-conversion HPLC method. These compounds were determined with the average recovery rate of 83.9% or more. The average total content of vitamin B6 forms was 1.0160.32 (mmol/L). Pyridoxal and pyridoxal 5′-phosphate were found in all of the samples, and their average contents were 0.7160.28 (mmol/L) and 0.1660.07 (mmol/L), respectively. Pyridoxamine, pyridoxine, pyridoxamine 5′-phosphate, pyridoxine 5′-phosphate, and pyridoxine-b-glucoside were found in 15, 14, 13, 9, and 7 samples, respectively. The presence of pyridoxine 5′-phosphate was for the first time found in human milk. A method for the determination of 4-pyridoxic acid, which is the excretion form of vit...