We reported previously the separation of ergot alkaloids by paper chromatographic method. The present paper deals with the quantitative determination of these alkaloids upon elution of the different spots and subsequent reaction with p-dimethylaminobenzaldehyde reagent. For the separate determination of ergonovine and total water-insoluble alkaloids, impregnate the paper with pH 4.4 buffer :formamide :acetone (2:1:1) mixture. Use chloroform as the developer, and let it run off the paper. Collect the organic phase in a small beaker beneath the paper, add 2 ml 0.2N H₂SO₄ and 4ml reagent, shake vigorously and determine colorimetrically the total water-insoluble alkaloid content. To determine ergonovine, cut off the paper holding this alkaloid, immerse in 2 ml 0.2 NH₂SO₄, stir well, add 4 ml reagent after 20 minutes and determine in the usual manner. For the determination of ergotamine and ergotoxine, the procedure was the same as above, except that a pH 2 buffer was used instead of pH 4.4. Ergotoxine ran off the paper while ergotamine remained on. Both were determined by treating with pdimethylaminobenzaldehyde solution as described above. The results were in agreement with those obtained by column chromatographic method. An unknown spot was found when pH 4.4 buffer was used; it also reacted with p-dimethylaminobenzaldehyde to a blue color. The R_f value was higher than that of ergonovine.