SAFETY TESTING OF HAIR DYE PREPARATIONS 751 TABLE II Primary Irritation Scores--Prolonged (O-hour) Daily Contact with P.P.D.-R. Dye q- H202 Multiple of Hmnan Rabbit Dose No. Days 14 21 Scores 0 1 0 0 0 0 2 0 0 0 0 3 0 0 0 0 Ix 1 0 2 2 2 2 0 2 2 2 3 0 2 2 2 3x 1 0 2 2 2 2 0 2 2 2 3 0 2 2 2 10x I 0 Died 2 0 2 2 2 3 0 2 2 2 thoroughly towel-dried, and a warm air stream, from a commercial 300- watt heater equipped with a fan, was directed over the rabbits for approximately five minutes. This procedure was repeated after each daily application of the test material. The effectiveness of this two- stage drying was demonstrated by the significantly reduced incidence of upper respiratory disease during these long test periods. One-half the animals were sacrificed 24 hours, and the remainder 14 days, after the last application no further treatment was given during these terminal periods. Prior to starting treatment, a base line hemogram including hemo- globin concentration, hematocrit, platelet, and total and differential leukocyte counts was obtained on each animal. In addition, semi- quantitative determinations were made for specific gravity, pH, glucose, protein, and microscopic examination of the sediment of urine. These were repeated prior to termination after the last application. All animals that died and half the survivors, which were sacrificed after 20 applications, were autopsied. Nine tissues, lung, heart, liver, kidneys, spleen, pancreas, thyroid, gonads, and marrow were examined micro- scopically. The procedure was repeated on the survivors 14 days later, and target organs were examined. Data obtained by this procedure were compared to a series in which the typical Draize procedure was followed. In this series, a com- mercially available oxidation hair dye similar to that used in the modi- fied procedure was used. Groups of 10 rabbits (evenly divided as to

TABLE III Primary Irritation Scores 1-Hour Contact with Graded Concentration H202 H202, % Rabbit No. Application No. 3 6 9 12 Males 1 1 0 1 1 1 2 0 0 0 2 3 0 0 0 1 4 1 1 2 1 5 1 1 2 1 6 H H H H 2 1 1 1 1 1 2 0 0 0 0 3 0 0 0 0 4 1 i i 1 5 H H H H 6 H H H H 3 1 1 1 1 2 2 0 1 i 1 3 0 2 i 2 4 2 2 2 2 5 i ! i 2 6 i 1 1 1 4 i i 2 2 2 2 0 i 2 1 3 i 1 i 1 4 1 i i 2 5 H H H H 6 H H H H 5 1 I 0 1 2 2 t) 1 1 2 3 1 2 2 2 4 1 i 2 2 5 H H H H 6 H H H H 6 1 1 0 2 2 2 0 ! 0 2 3 1 2 2 2 4 2 2 2 2 5 i 1 i 2 6 H H H H 7 1 0 0 0 1 2 1 1 1 1 3 1 1 2 2 4 i i 1 1 5 i i i 1 6 H H H H g 1 1 1 2 1 2 1 1 1 2 3 0 2 1 2 4 2 2 2 2 5 i 1 i 1 6 H H H H H = Indicates that hair regrowth was intense. No reading was made.