SAFETY TESTING OF HAIR DYE PREPARATIONS 761 indicative of the presence of blood per se or some foreign protein asso- ciated with injury, electrophoretic separations were made. The flow diagram for the procedure is shown in the scheme on page 760. No evi- dence of any unnatural or foreign protein was observed. The presence of hemoglobin protein was demonstrated by direct comparisons with an external standard. DISCUSSION AND SUMMARY The modified procedure utilizing the one-hour application resulted in a marked diminution or absence of the characteristic cracking, thick- ening, sloughins, and necrosis so often seen with the original Draize procedure and which do not contribute to the safety evaluation. This was in large part due to the shampooing and drying of the rabbits after each daily exposure, thus avoiding drying of the residual test material. No significant differences were observed between the two methods being evaluated as toxicological tGols. Not withstanding the similarity of findings with those of the Draize method, the one-hour procedure has distinct advantages with respect to utility and extrapolation to hu- mans. Though both procedures yield comparable results, the one- hour contact more closely resembles use conditions with loss of sensi- tivity as a test method. Both procedures, though valuable in ascer- taining irritation potential, failed to yield the means whereby more subtle differences between test preparations can be ascertained. A revised method whereby the hemoglobin content of 4 mm. skin plugs taken from the application sites was determined by an alkaline hematin method permitted differentiation between graded concentra- tions of a hydrogen peroxide solution. Irritation and inflammation resulting from this contact is associated with the extravasation, through the capillaries, of red cells and their collection at these inflammatory sites. Electrophoretic identification of this material as hemoglobin was made. The value of the modified method as a means of differen- tiating potentially irritating materials is indicated by the comparison of commercial oxidation hair dyes and their common vehicle. (Received May 20, 1965) REFERENCES (1) Draize, J. H., Appraisal of the Safety of Chemicals, Foods, Drugs and Cosmetics. Associa- tion of Food Drug Officials of the United States, Editorial Office, Baltimore, Md. (1959), p. 46. (2) E. Asch½im, Am. J. Physiol., 206• 327 (1964).
7•2 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS CHANGE OF ADDRESS (1) Allow 6 weeks to make the change. (2) Send change to Editorial Assistant, 761 North Valley Chase Rd., Bloomfield Hills, Michigan 48013 (3) Print name and new ad- dressmincluding postal zone number. Give old address--if possible re- turn addressed portion of the envelope in which your last Journal was mailed.
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SAFETY TESTING OF HAIR DYE PREPARATIONS 761 indicative of the presence of blood per se or some foreign protein asso- ciated with injury, electrophoretic separations were made. The flow diagram for the procedure is shown in the scheme on page 760. No evi- dence of any unnatural or foreign protein was observed. The presence of hemoglobin protein was demonstrated by direct comparisons with an external standard. DISCUSSION AND SUMMARY The modified procedure utilizing the one-hour application resulted in a marked diminution or absence of the characteristic cracking, thick- ening, sloughins, and necrosis so often seen with the original Draize procedure and which do not contribute to the safety evaluation. This was in large part due to the shampooing and drying of the rabbits after each daily exposure, thus avoiding drying of the residual test material. No significant differences were observed between the two methods being evaluated as toxicological tGols. Not withstanding the similarity of findings with those of the Draize method, the one-hour procedure has distinct advantages with respect to utility and extrapolation to hu- mans. Though both procedures yield comparable results, the one- hour contact more closely resembles use conditions with loss of sensi- tivity as a test method. Both procedures, though valuable in ascer- taining irritation potential, failed to yield the means whereby more subtle differences between test preparations can be ascertained. A revised method whereby the hemoglobin content of 4 mm. skin plugs taken from the application sites was determined by an alkaline hematin method permitted differentiation between graded concentra- tions of a hydrogen peroxide solution. Irritation and inflammation resulting from this contact is associated with the extravasation, through the capillaries, of red cells and their collection at these inflammatory sites. Electrophoretic identification of this material as hemoglobin was made. The value of the modified method as a means of differen- tiating potentially irritating materials is indicated by the comparison of commercial oxidation hair dyes and their common vehicle. (Received May 20, 1965) REFERENCES (1) Draize, J. H., Appraisal of the Safety of Chemicals, Foods, Drugs and Cosmetics. Associa- tion of Food Drug Officials of the United States, Editorial Office, Baltimore, Md. (1959), p. 46. (2) E. Asch½im, Am. J. Physiol., 206• 327 (1964).
7•2 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS CHANGE OF ADDRESS (1) Allow 6 weeks to make the change. (2) Send change to Editorial Assistant, 761 North Valley Chase Rd., Bloomfield Hills, Michigan 48013 (3) Print name and new ad- dressmincluding postal zone number. Give old address--if possible re- turn addressed portion of the envelope in which your last Journal was mailed.

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