The fractional determination of PAL and PIC was described and the effects of various conditions were reported. PIC is adsorbed on anionic exchanger and the non-adsorbed PAL is adsorbed on cationic exchanger. PIC eluted is determined as its lactone. PAL eluted is oxidized by ammoniacal silver to PIC which is determined as its lactone. For extracting PAL from tissues, direct heating of the homogenate in 0.1N H2SO4 at 130° for 1 hour was found to be most suitable. The values of 3 components of vitamin B6 in animal and plant tissues obtained by our method were reported and compared the total B6 values with those reported in the literature. They agreed fairly well, indicating the satisfactory specificity of the methods.