PHTHALATE ESTERS IN COSMETICS 459 Although phthalate esters have been commercially used for over 50 years, there are only limited human data on the relationship between exposure to phthalates and human health effects (9). It is also diffi cult to generalize toxic effects observed in different studies with different study protocols, phthalate esters, and animal models. Nevertheless, some parties have expressed concern about phthalate esters as an entire class and have advocated for their elimination from all consumer products (15). These concerns have been further ele- vated by studies documenting the presence of phthalate esters and/or their metabolites in human blood and urine samples (10,13). Public interest groups, including the Environ- mental Working Group (EWG), continue to allege that phthalate esters used as cosmetic ingredients pose a health hazard to humans and should be banned from cosmetic products (16). The European Commission (EC), after consultation with the Scientifi c Committee on Cosmetics and Non-food Products (SCCNFP), also recently classifi ed dibutyl phthalate and diethylhexyl phthalate as category 2 reproductive toxins and prohibited their use in cosmetics in the European Union (EU) (17,18). As part of its ongoing efforts to monitor phthalate ester use, the FDA conducted a survey of consumer cosmetic products beginning in 2002 for the presence of dimethyl, diethyl, benzylbutyl, dibutyl, and diethylhexyl phthalate esters in 48 consumer cosmetic products, including hair care products, deodorants, lotions and creams, nail products, fragrances, and body washes, purchased from local stores in the Washington, D.C. area. This initial product survey of cosmetics was undertaken to verify levels of phthalate esters reported by a coalition of environmental and public health organizations, the EWG, which advo- cated the removal of phthalate esters from cosmetic products (19), and to collect addi- tional data on phthalate ester levels in other cosmetic products. The analytical method chosen for the initial and follow-up studies was originally developed in our laboratory for the analysis of phenol, resorcinol, salicylic acid, and α-hydroxy acids in cosmetic products and salon preparations (20). The analytical method utilized a Celite column for sample clean-up followed by reversed-phase HPLC separation using gradient elution, and allowed rapid and sensitive quantitation of phthalate esters in con- sumer cosmetic products (6). Using this method, we determined that 67% of the prod- ucts surveyed contained at least one phthalate ester, while hair sprays, deodorants, nail products, and hair mousse contained two or more phthalates. The highest phthalate ester concentrations were found in nail products, with levels observed up to 60,000 μg/g. Diethyl phthalate was the most common phthalate ester found (27 out of 48 products). Dibutyl phthalate was found in ten products, while diethylhexyl phthalate was not found in any products tested. These results were consistent with those reported by the EWG in 2002 (19). The current study is a follow-up survey to determine if any of the products analyzed in our initial survey have been reformulated to reduce or eliminate phthalate esters. EXPERIMENTAL COSMETIC PRODUCTS A total of 84 adult-use and baby-care cosmetic products were purchased from local stores in the Washington, D.C. area or from the Internet. Among the 60 adult-use samples, 24 were nail enamel products, 12 were antiperspirants or deodorants, 11 were perfumes,

JOURNAL OF COSMETIC SCIENCE 460 seven were hair products, three were lotions, two were body washes, and one was a shampoo. The products selected included products analyzed in the previous 2002 survey that had high levels of phthalates to see if any had been reformulated to reduce/remove phthalates. In addition, since nail enamels had the highest phthalate levels in the original survey, 40% of the products analyzed were nail enamels. In response to a recent report regarding possible infant exposure to phthalate esters (21), baby-care products were included in our survey. Of the 24 baby-care products, 13 were shampoos and body washes and 11 were creams, lotions, and oils. REAGENTS AND MATERIALS The following reagents and materials were used: Hexane was purchased from Burdick & Jackson (Muskegon, Michigan). Acetonitrile and methanol were purchased from T. J. Baker (Phillipsburg, New Jersey). Fisher Scientifi c (Fairlawn, New Jersey) provided 2-propanol. All solvents were HPLC grade. Phthalate esters DMP (99%), DEP (99.5%), and BBP (98%) were purchased from Sigma Aldrich (Milwaukee, Wisconsin). DBP (≥98%) and DEHP (99.5%) were purchased from Sigma Aldrich. De-ionized water was prepared with a Milli-Q purifi cation system from Millipore (Billerica, Massachusetts). Celite 545 was purchased from Fisher Scientifi c. The extraction tubes and fi lter disks were obtained from Supelco (Bellefonte, Pennsylvania). Chromatographic separation was achieved using a Partisil ODS-3, 5-μm guard column (7.5 mm by 4.6 mm ID) and a Partisil ODS-3, 5-μm, analytical column (250 mm by 4.6 mm ID), both obtained from Altec Chroma- tography (Deerfi eld, Illinois). PHTHALATE ESTER CALIBRATION STANDARDS A primary standard solution of a mixture of the fi ve phthalate esters (∼1 mg/ml each) was prepared by adding approximately 100 mg of each to a 100-ml amber volumetric fl ask and diluting to the mark with hexane. Because of the wide range of possible concentra- tions in cosmetic products, three sets of working standards were prepared. One set was prepared at approximately 0.001, 0.003, 0.006, and 0.01 mg/ml by appropriate serial dilution of the stock solution. Similarly, a second set was prepared at approximately 0.01, 0.03, 0.06, and 0.1 mg/ml, and a third set for BBP, DBP, and DEHP only was prepared at 0.10, 0.30, 0.60, and 1.00 mg/ml. SAMPLE PREPARATION The extraction and analytical method previously developed by the FDA was used in this study (6). Briefl y, approximately 1 g of each test portion was weighed into a 40-ml beaker, mixed thoroughly with about 3 g of Celite, and then transferred to a 15-ml extraction tube. The sample/Celite mixture was covered with a fi lter disk and was compacted fi rmly with a stirring rod, and then hexane was passed through the fi lter disk by gravity fl ow. The extract was collected in a 10-ml volumetric fl ask until the volumetric fl ask had been fi lled to the mark. For solid deodorants, the sample was fi rst dispersed in hexane to create slurry