ANTIPOLLUTION COSMETIC EFFECTIVITY AGAINST AIR POLLUTANT ABSORPTION 35 METHODS AND MATERIALS REAGENTS AND MATERIALS Benzene, to luene, ethylbenzene, o- xylene, m-xylene, p-xylene, chlorobenzene, nitroben- zene, 1,2-dichloroethane, bromodichloromethane, 1,2-dibromoethane, bromoform, and naphthalene were provided by Scharlau (Barcelona, Spain) and Sigma-Aldrich (St. Louis, MO). Toluene-d8 was provided by Sigma-Aldrich, and it was used as internal standard. Standards and working solutions were prepared in acetone (GC analysis grade) obtained from Scharlau. Strat-M® membranes were purchased from Millipore (Temecula, CA). Buffer salts were obtained from Scharlau. COSMETIC PRODUCTS COMPOSITION Antip ollution cosmetic A is a daily skin care cream (viscous emulsion) that protects from sunlight and blue radiation, preventing skin photoaging. This cosmetic product fi rms, smoothes, and reduces wrinkles, due to paracress plant extract. It is suited to sensitive skin because of the presence of active principles such as licorice extract and bioactive molecules from stem cells from cotton. Antipollution cosmetic B is also a rich texture cream (viscous emulsion) that welfares and pro- tects the skin against external agents such as contamination, sun radiation (infrared, ultravio- let, and blue light), and oxidant agents, increasing skin fi rmness and elasticity while intensely moisturizing. These properties for treating and preventing adverse environmental effects are provided by its combination of active principles including vitamin C, peptides (carnosine and oligopeptide-1), alantoin, panthenol, and bioactive molecules from stem cells from cotton. PHYSICOCHEMICAL PROPERTIES OF DEVEL OPED COSMETIC PRODUCTS Physicochemical properties such as viscosity, density, pH, conductivity, and refraction index of developed cosmetic products were determined following the procedures de- scribed next. Viscosity was measured using a DV-III Ultra rotational viscometer from AMETEK Brookfi eld (Middleboro, MA). The measurement was carried out with a rota- tional speed of 50 rpm and read after 3 s after starting the viscometer. An electronic densimeter MDS-300 from Alfa Mirage (Tokyo, Japan) was used for density determina- tion. A 3-mL syringe was fi lled with sample. The cell was fi lled with the sample until it fl ows out by the other exit (2 mL of sample, approximately). The pH values of the respec- tive specimen were measured by means of the PHS-3B pH meter from TBT (Nanjing, China). The values of electrical conductivity were measured using an EC-meter GLP-31 from Crison (Barcelona, Spain). The refraction index was measured using an RE40D Re- fractometer from Mettler Toledo (Barcelona, Spain). In all the experiments, sample tem- perature was 25°C, and the measurement was repeated three times for the sample, the fi nal result being an average of three independent measurements. SIMULATION CHAMBER A modifi ed closed enclosure model HZ 08252, with 640 L internal volume, from Bruker (Billerica, MA) was used as a simulation chamber (see Figure SM1 of Supplementary

JOURNAL OF COSMETIC SCIENCE 36 Material). Clean and dry air at a fi xed fl ow rate of 2.7 L min-1 was introduced in the cham- ber using an MPB1200 rotameter from MPB Industries (Kent, United Kingdom), cali- brated by a high-volume bubble fl owmeter. Clean air was spiked with HAPs by using an isocratic LC pump HP 1,050 series from Hewlett-Packard (Palo Alto, CA) adjusted to 5 μL min-1 and a glass T connector (see Figure SM2 of Supplementary Material). Air homo- geneity inside the chamber is obtained with two fans with vibration-dampening rubber corners. Infi nite dose conditions were maintained during all the experiments. The simu- lation chamber was placed in a-50 m3 closed room, with controlled temperature (25 ± 1°C) and a fume hood system. DETERMINATION OF HAPS IN THE SIMULATION CHAM BER BY ACTIVE SAMPLING HAP concentration inside the chamber was mea sured by active sampling using a low- volume personal air sampling TUFF Standard from Casella measurements (Bedford, United Kingdom), operating with a low-fl ow adaptor at a fl ow rate of 76 mL min-1 for 30 s. Glass thermal desorber (TD) tubes, capped with perfl uoroalkoxy-polytetrafl uoroeth- ylene (PFA-PTFE) ferrules, were obtained from Perkin Elmer (Waltham, MA) and fi lled with 150 mg of Tenax TA (35–60 mesh) provided by Alltech (Selmsdorf, Germany). Tenax was conditioned before sampling at 300°C during 2 h. All Tenax tubes were analyzed before sampling to demonstrate the usefulness of the conditioning procedure. Active sampling pump fl ow was regulated using an ADM calibrated fl owmeter (Agilent Technologies, Palo Alto, CA) before each sampling. After sampling, tubes were capped with PFA-PTFE ferrules and stored at -10°C until analysis. TD tubes were ther- mally desorbed using a Turbo Matrix series TD from Perkin Elmer coupled to a Trace GC-Polaris Q gas chromatography–mass spectrometry (GC-MS) detector from Finnigan (Waltham, MA), equipped with an Agilent HP-5MS capillary column (30 m, 0.25 mm, and 0.25 μm). Thermal desorption was carried out at 260°C for 20 min using a 75 mL min-1 helium fl ow rate, and desorbed analytes were transferred to a Tenax cold trap at -10°C. A quick trap desorption was carried out at 270°C at 99°C s-1, and the analytes were desorbed and directly transferred to the chromatographic column using a transfer line temperature at 275°C, using a split fl ow of 1:15. The used chromatographic conditions started at an initial temperature of 40°C, held for 8 min, increased at a rate of 20°C min-1 up to 200°C, and fi nally held for 2 min. Acquisition were performed in full scan mode using a mass range from 50 to 150 m/z. A calibration curve was prepared by the direct addition of 10 μL H AP standards in ace- tone directly inside Tenax packed TD tubes. Curve was prepared at fi ve concentration levels ranging from 0.005 to 1.0 μg target analytes. Then, 10 μL toluene-d8 standard solution in acetone at a concentration of 0.01 g L-1 was added inside the Tenax tube to obtain a fi nal mass of 0.1 μg, as internal standard. ABSORPTION STUDIES IN THE SIMULATION CHAMBER In vitro dermal absorption studies were performed using modifi ed vertica l diffusion cells, following the Franz method (10,11). In this study, the donor compartment was