ASSESSMENT OF OCULAR IRRITANCY 321 experienced an adverse event. This event, which occurred prior to test material instil­ lation, was unrelated to the test material or to the study procedures. Other evidence of the safety of this methodology is provided by the repeated willingness of subjects to enroll in these studies as well as the resolution of all observed ocular irritation during the course of the studies. SUBJECTIVE IRRITATION Average maximum score levels of subjective irritation (Figure 2), including stinging, burning, itching, dryness, and/or foreign body sensation, demonstrated that mascara and powder eye shadow exhibited mild to moderate irritation (2.8 ± 0.102 and 2.45 ± 0.182, respectively), which was statistically higher (p 0.05) than for the remaining product types, except for baby wash and eye makeup remover as compared to powder eye shadow. Liquid makeup with an average maximum score level of 0.5 ± 0.088 elicited significantly lower levels of reported subjective irritation than all other products (p 0.05). OBJECTIVE IRRITATION (SLIT LAMP BIOMICROSCOPE EXAMINATION) Objective ophthalmic evaluation consisted of examination of lacrimation, eyelid inflam­ mation, palpebral and bulbar conjunctiva! inflammation, and corneal abnormalities. No lacrimation, eyelid inflammation, or corneal abnormalities were observed for all product types. The average maximum score levels of combined palpebral and bulbar conjunctiva! inflammation scores (maximum combined score of 8) were significantly higher (p 0.05) for shampoo and baby wash, with scores of 4.075 ± 0.126 and 4.1 ± 0.16, respectively, than for liquid makeup, eye make�p ·remover, mascara, powder eye shadow, and facial 4 3.5 " 3 .J t 2.5 8 2 ,,, G) ! 1.5 G) 0.5 0 Liquid makeup Shampoo Baby wash Eye makeup remover Mascara Powder eye Facial cleanser shadow Figure 2. Average maximum levels of subjective irritation.

322 JOURNAL OF COSMETIC SCIENCE cleanser, with scores of 3.025 ± 0.12, 2.783 ± 0.141, 1.8 ± 0.25, 3.4 ± 0.11, and 2.34 ± 0.25, respectively (Figure 3). DISTRIBUTION OF FLUORESCEIN STAINING The maximum area and density scores at each examining timepoint for the fluorescein staining patterns of each ocular tissue were multiplied and totaled for all examination intervals. Scores from each evaluated tissue were weighted, based on a modification of the Draize method (5,6) for assessing ocular irritancy potential, and totaled to determine an overall fluorescein staining score for all eyes by the following equation: Total weighted score = (palpebral + bulbar) x 2 + cornea x 5 + caruncle x 1. The distributions of the average weighted score (Figure 4) for liquid makeup, shampoo, baby wash, eye makeup remover, mascara, powder eye shadow and facial cleanser were 11.4 ± 1.34, 31.03 ± 1.52, 39.19 ± 2.70, 9.6 ± 0.91, 39.92 ± 3.45 and 9.55 ± 1.98, respectively. The scores for shampoo, baby wash, and powder eye shadow were statistically signifi­ cantly higher than those of the others (p 0.05). POST-INSTILLATION EVALUATION INTERVALS The Draize method and the modified Draize methods, such as the FHSA method, the OECD method, and the FIFRA/TSCA method (7 ,8), established the stipulated intervals for observing irritation, which were limited to 1, 24, 48, and 72 hours after adminis­ tration of the test material in animals. Human eyes are highly sensitive to cosmetic products and respond frequently and quickly to test material exposure. Therefore oph­ thalmic evaluations were performed at the following time intervals: 30 seconds, 5 minutes, 15 minutes, 60 minutes, 120 minutes, and 24 hours post-instillation. Sub- 4.5 4 3.5 (I) : 3 � 2.5 "' 2 a, 1.5 0.5 0 Liquid makeup Sha"l)OO Baby wash Eye makeup remowr Mascara PO'Ader eye Facial cleanser shadow Figure 3. Average maximum levels of objective irritation.