The vitamin B6 status of high school students and its relationship with dietary intake were investigated in this cross-sectional study by face-to-face interview. A total of 157 healthy students aged 16–18 years (eighty-three boys and seventy-four girls) were randomly recruited from two out of nineteen senior high schools in Tainan, Taiwan. Vitamin B6 intakes were calculated from three 24-h dietary records. Direct and indirect vitamin B6 status indicators were measured in plasma, erythrocytes and urine. The anthropometric data, being similar to those of the first Nutrition and Health Survey in Taiwan (1993–1996), showed the normal growth and development of these students. All students except one girl (28·7 nmol/l) had plasma pyridoxal-5′-phosphate (PLP) levels >35 nmol/l, indicating an adequate vitamin B6 status. The mean dietary vitamin B6 intakes of boys and girls were 1·04 (sd 0·29) and 0·96 (sd 0·27) mg/d, respectively. Vitamin B6 status indicators, including plasma PLP, erythrocyte alanine aminotransferase activity coefficient (EALT-AC), aspartate aminotransferase activity coefficient (EAST-AC) and urinary 4-pyridoxic acid (4-PA), were correlated with vitamin B6 intake. Students with adequate values of plasma PLP (>35 nmol/l), EALT-AC ( < 1·25), EAST-AC ( < 1·8) and urinary 4-PA (>3·0 μmol/d) had median intakes of 1·08 and 1·01 mg/d, respectively, for boys and girls. This study suggests that vitamin B6 requirements for boys and girls aged 16–18 years were approximately 1·1 and 1·0 mg/d, respectively.