Some of the guanidium groups in the solution might be destroyed by the hypochlorite, resulting in difficulties in testing results.The test rate of Sakaguchi’s reaction is very slow and therefore, quantitative analysis of colored products is very much difficult with this test.Negative Test: A negative Sakaguchi result is indicated by absence of red color, and in this case, this confirms absence of arginine or a guanidinium compound.This confirms the presence of an arginine or guanidinium compound. Positive Test: A positive Sakaguchi test result is indicated by the formation of red color. STEP 2 TILT THE TUBEAND CAREFULLY POUR ALONG ONE SIDE OF THE TUBE 1.0mL OF CONCENTRATED STEP 3 A REEDISH VIOLET RING WILL FORM AT THE JUNCTION BETWEEN THE TWO LAYERS WHICH PROVES THE PRESENCE OF TRYPTOPHANSTEP 1 ADD 2.0 mL OF HOPKINS- COLE REAGENT TO 2.0 ML OF EGG WHITE SOLUTION IN A TEST TUUBE.Observe the test tube for development of color.Add 4 to 5 drops of bromine water or add 1 to 2 drops of 10% sodium hypochlorite.Then, add two drops of 1-naphthol and mix thoroughly.Mix 1ml of 40% NaOH with 3ml of the amino acid or test solution in a test tube.This should be done in a fume chamber or use 10% sodium hypochlorite solution. Bromine water: Add a few drops of bromine to 100 ml water and shake.Guanidines: 0.1% solutions of glycocyamine, methyl guanidine and creatine.Protein solution: Egg albumin in distilled water (around 10%). Amino acids: 0.1% solution of amino acids like glycine, arginine, lysine, tyrosine etc.
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