1 2 3 4 5

Simple method for the determination of lead in lipstick treated with tetramethylammonium hydroxide by graphite furnace atomic absorption spectrometry

Aline Rodrigues Soaresa, Clsia Cristina Nascentesa*

6Instituto de Cincias Exatas, Departamento de Qumica, Grupo de Espectrometria Atmica 7e Preparo de Amostras (GEAPA), Universidade Federal de Minas Gerais, Avenida 8Presidente Antnio Carlos, 6627, Pampulha, PO Box 702, CEP 31270-901, Belo 9Horizonte, Minas Gerais, Brazil 10 11 12 13 14 15 16 17 18 19 20 21 22 23Abstract 1 2
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*Corresponding author: clesianascentes@yahoo.com.br Tel: + 55 (31) 3409 5759.

24 25 26atomic A simple method for determining total lead in lipstick samples by graphite furnace absorption spectrometry (GF AAS), after sample treatment with

27tetramethylammonium hydroxide (TMAH) was developed. Multivariate optimization was 28used to establish the optimal conditions of sample preparation. The GF AAS temperature 29program was optimized through pyrolysis and atomization curves. An aliquot containing 30approximately 50 mg of the sample is simply mixed with TMAH and heated in a water bath 31at 60 C for 60 min. Using Nb as the permanent modifier and Pd as the chemical modifier, 32the optimal temperatures were a 900 C and 1800 C to pyrolysis and atomization,

33respectively. Under optimum conditions, the linear range was from 0.62 to 50.0 g L 1, 34with a detection and quantification limits of 0.04 and 0.12 g g -1, respectively. The relative 35standard deviation varied from 1.54 to 4.20%. The recovery rate was from 94.1 to 109%, 36and no significant differences were found between the results obtained with the proposed 37method and the microwave decomposition method for real samples. Lead was detected in 38all tested lipstick samples in the range of 0.17 to 4.54 g g-1. 39 40 41 42 43 44Keywords: lead; lipsticks; graphite furnace; TMAH; multivariate optimization 45 46 3 4 1. Introduction
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47 48 Over time the use of cosmetic products worldwide is increasing at an alarming rate

49due to the everlasting pursuit for individual beautification and due to a sharp rise in product 50advertisements in the electronic media. Most of these cosmetic products are directly applied 51to the human skin. While the skin provides a great protective barrier, some of the 52ingredients in cosmetic products can penetrate the skin and reach vital internal organs via 53the systemic circulation [1]. Cosmetic products which are applied to mucous membranes 54are even more hazardous, as in the case of lip products such as lipsticks. In addition to these 55risks, lipsticks also have the higher risk of direct oral ingestion, aggravating the negative 56effects of its chemicals [2]. 57 Recent media reports described the presence of Pb in lipsticks and it suggest that

58under conditions of ordinary use, the potential Pb exposure is harmful [35]. It has been 59estimated that a woman inadvertently ingests 1.8 kg of lipstick in a lifetime [6]. Thus, it 60stands to reason that when a woman licks her lips, eats and drinks while wearing lipstick, 61she can conceivably ingest Pb from the lipstick. With the build-up of Pb in the body over 62time, exposure levels and consequences can be significant. Lead may cause serious health 63hazards such as both acute and chronic poisoning, pathological change of organs and 64diseases related to cardiovascular, kidney, bone, and liver, and they can even cause cancer 65owing to excessive accumulation in human body [7, 8]. Lead has also been linked to 66miscarriage, and reduced fertility in both men and women. In pregnant women, lead can 67enter the fetal brain through the placenta [9]. 68 Lead exposure assessments are frequently based on intake from food, water, or air.

69Toxic effects are usually due to long term exposure. [10]. The World Health Organization 5 6
3

70(WHO) [11] estimated a range of 4.010 g/day total intake from air and water in adults. 71The major source of lead for nonoccupationally exposed adults is food, with a range of 23 72500 g/day total lead intake. Lead in lipsticks represents a minor source of lead exposure 73compared to other one because the amount of lipstick applied daily is small. Nonetheless, 74one should not exclude the fact that lead accumulates in the body over time and continuous 75lead-containing lipstick application can lead to significant exposure levels [12]. 76 Recently, the Campaign for Safe Cosmetics (CSCs) in the United States raised

77another concern about the presence of lead in lipsticks. They found that more than half of 78the tested 33 brand-name red lipsticks (61%) contained Pb in the range of 0.030.65 g g -1 79[6]. Lead contamination of lipsticks may originate from Pb solder or leaded paint in 80production equipment or from contaminated dust [3]. Lipsticks also may be contaminated 81with Pb if they are manufactured with ingredients that naturally contain Pb or are produced 82under conditions that could introduce Pb into the ingredients. Dyes and pigments used as 83ingredients in lipsticks are regulated as color additives by the FDA and must undergo 84premarket approval by the agency before they may be used in any cosmetics [13]. In 85current regulations, most color additives approved for cosmetic use are permitted to contain 86up to 20 g Pb/g [14]. 87 Different analytical techniques have been used for the determination of lead in

88lipsticks and other cosmetics, such as laser induced breakdown spectroscopy (LIBS) [1], 89flame atomic absorption spectrometry (FAAS) [15], inductively coupled plasma mass 90spectrometry (ICP-MS) [13], inductively coupled plasmaoptical emission spectrometry 91(ICPOES) [16] and graphite furnace atomic absorption spectrometry (GF AAS) [12]. G 92GF AAS appears to be a good alternative for the determination of trace elements, such as 7 8
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93lead, in lipsticks, because it is one of the most sensitive techniques with limits of detection 94in the range from g L1 to ng L1 and it is extremely tolerant to complex matrices [17]. 95 Complex samples, such as lipsticks, require conversion to a form compatible with

96the used instrumentation, allowing, if possible, a simple and effective calibration [18]. In 97the works described in literature [13, 15], the sample preparation involved a acid digestion 98in microwave oven. However, acid digestion requires the use of strong acids, in 99disagreement with the green chemistry principles and is usually time consuming, even if 100assisted by microwaves. In addition, it is subjected to analyte loss and/ or sample 101contamination [19]. In this context, the alkaline solubilization of samples with 102tetramethylammonium hydroxide (TMAH) is also an interesting alternative which has 103being employed with success in biological samples such as milk powder, bovine muscle, 104mussel tissue, fish muscle, human hair, human blood and nail samples prior to their 105analysis [2023]. Usually, the treatment is very simple and fast. 106 The aim of this work is the development of a simple analytical method for the

107determination of lead in lipsticks by GF AAS, after sample treatment with 108tetramethylammonium hydroxide. A multivariate optimization strategy based on a factorial 109and central composite design was employed. The method was validated and employed to 110quantify lead in several lipsticks samples. 111 112 1132. Experimental

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114 1152.1. Instrumentation 116 117 A Perkin Elmer AAnalyst 400 atomic absorption spectrometer (Norwalk, CT, USA)

118equipped with an HGA 800 graphite furnace and an AS-800 autosampler was used in all 119measurements of Pb integrated absorbance. The background correction was made with a 120continuous light source (deuterium lamp). Argon (99.996%; White Martins, So Paulo, SP, 121Brazil) was used as the purge gas. Perkin Elmer pyrolytic graphite-coated tubes with Lvov 122platforms were used. A Pb hollow cathode lamp (Perkin Elmer, Norwalk, CT, USA) was 123used at a wavelength of 283.3 nm, a slit width of 2.7/1.05 nm and a current of 10 mA, 124which were the manufacturers recommended conditions. 125 A Milestone Ethos 900-Mega II microwave oven (FKV Milestone, Milan, Italy)

126with a PTFE-vessel rotor was used to digest the lipstick samples. 127 128 1292.2. Reagents, materials and samples 130 131 Deionized water (resistivity of 18.2 M cm-1) was obtained with a Direct-Q system

132(Millipore, Billerica, MA, USA) immediately before use for the preparation of all solutions. 133The alkaline solubilisation was performed with tetramethylammonium hydroxide 25% (v/v) 134in water (Sigma-Aldrich, So Paulo, Brazil). Concentrated nitric acid and hydrogen 135peroxide were obtained from Merck (Darmstadt, Germany). Iridium, niobium, tantalum, 136ruthenium, rhodium and zirconium solutions (1000 mg L -1) were purchased from Fluka 11 12
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137(Buchs, Switzerland) in 1.0 mol L-1 hydrochloric acid. A 1000 mg L-1palladium solution 138was obtained from Ultra Scientific (North Kingstown, RI, USA). 139 Plastic bottles, autosampler cups, and glassware were all soaked in 20% v v -1 HNO3

140for 24 hours, rinsed several times with Milli-Q water, and dried at room temperature. An 141auto-sampler washing solution containing 0.05% v v -1 Triton X-100 (Merck, Darmstadt, 142Germany) and 0.1% v v-1 isopropanol (Sigma-Aldrich, So Paulo, Brazil) was used to avoid 143analyte adsorption onto the surface of the container and clogging of the capillary sampling 144tip. The lead stock solution (1000 mg L-1) was prepared using lead from Titrisol Merck 145(Darmstadt, Germany) in a 5% v v-1 nitric acid solution. 146 Lipsticks samples of different brands and colors were acquired at the local market

147(Belo Horizonte, Brazil). Selected samples were from China, France, USA and Brazil. 148 149 1502.3. Graphite tube treatment 151 152 The graphite tubes were treated independently with 500 g of each permanent

153modifier studied (Zr, Ir, Rh, Ru, Nb and Ta) by applying 25.0 L of each metal solution 154(1000 mg L-1) to the platforms, that were then submitted to a furnace temperature program 155as described previously [24]. This procedure was repeated twenty times. 156 157 1582.4. Optimization strategies 159 13 14
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160

The lipstick sample employed for optimization was previously analyzed to evaluate

161the analytical signal related to Pb concentration. As the integrated absorbance obtained was 162considered satisfactory, the lipstick sample used in this step was not fortified. 163 To establish the optimal conditions of sample preparation, a 2 4-1 factorial design was

164employed to evaluate the effect of the variables heating hold time (30 or 60 min); heating 165temperature (60 or 100 C), sonication time (0 or 30 min) and volume of TMAH (0.50 or 1661.00 mL). Based on the results, the optimal conditions were determined through a central 167composite design (CCD; 10 experiments, including 3 replicates in the center point) in 168which were studied the volume of TMAH and heating time. The GF AAS analyzes were 169carried out under the conditions recommended by the manufacturer Tabela 1. The data 170were processed by the Statistica 6.0 software. 171 Experiments were conducted to choose the appropriate permanent modifier for lead

172determination in the lipstick sample. The integrated absorbance measurement and the 173background signal for this sample were obtained using separate graphite tubes treated with 174permanent modifiers (Rh, Ir, Zr, Nb, Ta, Ru) and Pd as a chemical modifier (co-injection of 1752 L of 1000 mg L-1 solution). A tube without a permanent modifier was evaluated too. The 176results when using these modifiers can be seen in Table 2. 177 The best modifiers from this analysis were chosen, and the temperature program

178was optimized through pyrolysis (from 400 to 1100 C) and atomization (from 1500 to 1792200 C) curves. 180 1812.5. General procedure 182 15 16
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183

Aliquots containing approximately 50 mg of the lipstick sample were weighed in

184polypropylene tubes, followed by the addition of 460 L of a 25% w/v TMAH solution. 185Then, the volume was made up to 10 mL with water and the mixture was kept in a water 186bath at 60 0C for 60 min. 187 188 1892.6. Microwave-assisted acid digestion 190 191 The microwave-assisted acid digestion was used for comparison to the results

192obtained in the proposed method. A mass of 300 mg of lipstick sample was weighed into 193PTFE-vessels. A volume of 5.0 mL of HNO 3 and 2.0 mL of HF was added to each vessel, 194which was then submitted to the heating program of the microwave oven [13]. The 195program was as follows: the system was heated for 10 min to reach 180 C and this 196temperature was maintained for 30 min. After cooling, the samples were quantitatively 197transferred into graduated flasks and diluted up to 25.0 mL with high-purity, deionized 198water. The samples were digested in triplicate. 199 200 2013. Results and discussion 202 2033.1. Optimization of alkaline solubilization 204

17 18

205

The TMAH is a strong organic base, water soluble alcohols resulting colorless

206solutions with amine odor, and capable of complexing and stabilizing volatile elements. 207[25]. Tetramethylammonium hydroxide-based procedures provide an alternative to 208conventional acid digestion, since satisfactory accuracy, precision and instrument 209performance have been reported in the majority of such studies. A reduction in sample 210preparation time and the long-term stability of the digests are two of the greatest advantages 211of this reagent [26]. Treatment of lipsticks samples with tetramethylammonium hydroxide 212(TMAH) is also an interesting alternative, which has being employed with success in the 213solubilization of samples with high levels of fat, as biodiesel, bovine muscle, fish muscle, 214milk, and human blood prior to their analysis [2731]. This is the first time that treatment 215with TMAH is applied to lipsticks samples, prior to their analysis and so it is important 216optimize the variables that can affect the process of alkaline solubilization. 217 The optimal conditions of alkaline solubilization were evaluated using a 2 4-1 factorial

218design. The results of the factorial design are shown in a Paretos chart of the estimated 219effects (Figure 1). The volume of TMAH and heating time variables had significant effects 220on the response (integrated absorbance) at the 95 % confidence level. Based on these 221results, the effect of volume of TMAH was negative and effect of heating time was 222positive, indicating that lower volumes of TMAH and high heating times produced a final 223solution enough stable and homogeneous to allow its analysis by GF AAS. High volumes 224of TMAH led to a reduction of analytical signal and increased background. 225 Through the results of the factorial design, it was observed that the volume of

226TMAH and heating time needed for a final optimization. A response surface methodology 227using a central composite design (CCD) was used to determine the optimal conditions and 19 20
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228critical points for these variables. Been studied a range of 300 to 700 L for the volume of 229TMAH and a range of 30 to 90 minutes for heating time. The heating temperature and the 230sonication time showed no significant effects; their values were set at 60 0C and 0 min, 231respectively. The results of the experiments the central composite design of the array was 232generated a response surface (Figure 2). The optimal conditions were heating time of 60 233min and volume of TMAH 25 % m/v corresponding to 460 L. Ghisi et al. [27] used 500 234L of TMAH 25% w/v in the solubilization of 500 mg of biodiesel samples, keeping the 235mixture at 90 C for 5 min before completing the volume with water to 5 mL. 236 237 2383.2. Modifiers and graphite furnace temperature program 239 240 The difficulties due to sample complexity can be overcome with chemical

241modifiers. In many cases, the chemical modifiers may help predigest the matrix and 242simultaneously improve the sensitivity of the assay [32]. To choose the best permanent 243modifier, the analysis was performed in triplicate using the manufacturers recommended 244conditions for drying, pyrolysis and atomization steps. The pyrolysis and atomization 245temperatures used were 700 and 1800 0C, respectively. The lipstick sample used for choose 246the best permanent modifier, was subjected to alkaline solubilization procedure previously 247established. 248 The results obtained in this study are showed in Table 2. The highest analytical

249signal that had good precision and low background signal was obtained using Nb as the 250permanent modifier. The use of Pd as the chemical modifier allowed for an increase in the 21 22
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251analytical response and was used throughout the study (Figure 3). Dobrowolski et al. [33] 252studied the action of mixed permanent modifiers niobium (Nb)/iridium (Ir) and tungsten 253(W)/iridium (Ir) for cadmium and lead determination in sediments and soils by slurry 254sampling graphite furnace atomic absorption spectrometry. Application of Nb/Ir 255modification for determination of Cd and Pb by slurry sampling GFAAS resulted in 256prolong tube lifetime. It was pointed out that Nb/Ir is a favorable permanent modifier for Pb 257and Cd determination from the group of refractory metals. 258 After selecting the modifiers, the temperature program was optimized through

259pyrolysis and atomization curves (Figure 4). Optimal pyrolysis and atomization 260temperatures were established in 900 and 1800 0C, respectively. The atomization 261temperature selected for the lipstick sample was identical to that described by Baysal & 262Akman [30] for the determination of lead in hair (1800 0C). The value established for the 263pyrolysis by these authors was 800 0C, which is very close to the temperature set for lipstick 264sample in the proposed method. 265 266 2673.3. Figures of merit 268 269 Using the optimal experimental conditions, the analytical parameters for Pb

270determination in lipstick samples were obtained (Table 3). To verify the linearity and 271matrix effects three calibration curves using aqueous standard solutions with 2.5% w/v 272TMAH and three standard addition calibration curves were prepared. The comparison of 273the slopes of the curves by means of the F and t-student tests revealed significant 23 24
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274differences at the 95% confidence level. Therefore, standard addition calibration curves 275were used for calibration in all additional experiments. The limits of detection (LOD) and 276quantification (LOQ) were calculated, by IUPAC criterion, according to LOD=3 sblank/a and 277LOQ= 10 sblank/a, where sblank is the standard deviation of the blank (n=10) and a is the slope 278of the analytical curve. The values obtained for the detection and quantification limits were 2790.04 g g-1 and 0.12 g g-1, respectively, proving adequate to the proposed work. The 280linearity was estimated by constructing an standard addition calibration curve in the range 281of 0.62 to 50.0 g L-1, and a good adjustment was verified (r = 0.999). The sensitivity was 282evaluated calculating the characteristic mass (8.5 pg) and comparing it to the recommended 283values (9.0 pg). Comparatively, Damin et al. [35] determined Pb in crude oil samples by 284GFAAS obtaining a characteristics mass of 8.7 pg using Pd + Mg as the chemical modifier 285and Pereira et al. [36] obtained a characteristic mass of 8.3 pg in sediment samples using 286Zr as the permanent modifier. The precision was evaluated in terms of repeatability and 287intermediate precision. The repeatability, was determined by analyzing of 7 replicates a of 288the same sample spiked with 2.0, 10.0 and 20.0 g L-1 of the analyte. For determination of 289intermediate precision, the 7 replicates of the sample spiked (in the range of 2.0 to 20 g L 2901) were analyzed by the same analyst, same apparatus, under the same conditions of use, on 291different days (1 day, 1 week and 1 month). The relative standard deviation (RSD) was 292calculated and ranged from 2.01% to 4.72%. 293 294 Certified reference materials similar to lipsticks was not available. Thus, the

295accuracy of the method was checked by addition-recovery studies and by comparison with 296conventional acid digestion procedure. A recovery of 94.1% to 109.0% was obtained in 25 26
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297experiments with spiked samples (in the range of 2.0 to 20 g L -1). The results obtained by 298alkaline solubilization and the microwave-assisted digestion procedure (Table 4) were 299compared by paired t-test. The results obtained by both procedures were in agreement at a 30095% confidence level. 301 302 3033.4. Lead determination in lipstick samples 304 305 The lead concentration in 40 different brands and colors lipstick samples from

306several origins was determined by the proposed method (Figure 5). The concentrations 307ranged from 0.22 to 2.36 g g -1 for the beige colors, 0.17 to 3.38 g g -1 for rose colors, 1.62 308to 4.54 g g-1 for red colors and 1.46 to 4.28 g g-1 for brown colors. When we looked at the 309lead contents according to the color of lipsticks (Figure 5), it seems the highest lead content 310was found in red and brown lipsticks. The highest values we found corresponded to 311lipsticks imported from China, with lead levels ranging from 2.07 to 4.54 g g-1. 312 Lead was detected in all tested lipstick samples, the highest amount content was

3134.54 g g-1 and the lowest was of 0.17 g g-1, while the average was 1,78 g g-1. The 314samples showed concentration of lead below 20 g g 1, that is established as the maximal 315lead limit as impurities in color additives in the cosmetics for external use, formulated 316following the good manufacturing practices [37]. The level of lead was also less than 20 g 317g1 in 20 lipsticks tested by the US FDA [13], the highest amount content was 3.06 g g 1 318and the lowest was 0.09 g g1. Lead was also found in 61 per cent of the 33 brands of 319lipsticks tested by the CSC, with lead levels ranging from 0.03 to 0.65 g g-1 [3]. 27 28
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Although the lead content found in the lipstick samples represents a minor source of

321lead exposure compared to other sources such as water, food or air one takes, such 322exposure should still not be overlooked. Some of the ingredients in lipstick can penetrate 323the skin and reach vital internal organs via the systemic circulation. In addition to these 324risks, lipsticks also have the higher risk of direct oral ingestion, aggravating the negative 325effects of its chemicals [2]. The metals accumulate in the body over time and repetitive 326metal-containing product application can lead to significant exposure levels. The toxicity 327resulting from exposure to lead is well known; it can cause damage to the kidneys and the 328central nervous system and memory loss, among other symptoms [38]. 329 330 331 3324. Conclusions 333 334 A simple and fast analytical method for the determination of lead in lipsticks

335samples by GF AAS, after alkaline treatment with TMAH was proposed and validated. The 336alkaline sobulization avoids sample contamination and analyte loss, due to no acid 337digestion is required. Multivariate optimization was adequate for obtaining the optimal 338conditions of sample preparation. The temperature program was optimized through 339pyrolysis and atomization curves. Under these conditions, the accuracy, precision, and 340sensitivity of the method were adequate to quantify trace concentrations of Pb in lipsticks . 341The analyte concentrations found in the commercial samples were especially high for red

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342and brown lipsticks of brands imported from China. These results confirm the importance 343of the quality control of the cosmetics and the applicability of the proposed method. 344 345Acknowledgements 346 347 The authors are grateful to the Fundao de Amparo Pesquisa do Estado de Minas

348Gerais (FAPEMIG) and the Conselho Nacional de Desenvolvimento Cientfico e 349Tecnolgico (CNPq) by financial support. A.R.S is thankful to Coordenao de 350Aperfeioamento de Pessoal de Nvel Superior (CAPES) by fellowship. 351 352 353 354 355 356 357 358 359 360 361 362References 363

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364[1] M.A. Gondal, Z.S. Seddigi, M.M. Nasr, B. Gondal, J Hazard Mater 175 (2010) 726365732. 366 367[2] C.C. Wang, A.N. Masi, L. Fernndez, Talanta 75 (2008) 135140. 368 369[3] Campaign for Safe Cosmetics, A Poison Kiss: The Problem of Lead in Lipstick, 2007. 370Available from: http://www.safecosmetics.org/about/reports.cfm. 371 372[4] Dangerous Levels of Lead in Lipstick, Lip Gloss?, 2006. Available from: 373www.healthy-communications.com/6lipstickdangers.htm. 374 375[5] Is Lead Inside Lipstick, 2006. Available in: http://www.wpxi.com/news/news/is-lead376inside-lipstick/nGkgh/ 377 378[6] The Campaign for Safe Cosmetics, 2007, A poison kiss: the problem of lead in lipstick. 379Available from: http://www.safecosmetics.org/docUploads/A%20Poison%20Kiss.pdf. 380 381[7] K. Shrivas, K.D. Patel, J. Hazard. Mater. 176 (2010) 414417. 382 383[8] K. Koller, T. Brown, A. Spurgeon, L. Levy, Environ. Health Perspect. 112 (2004) 987384994. 385 386[9] C. Basheer, S.H. Tan, H.K. Lee, J. Chromatogr. A 1213 (2008) 1418. 387 33 34
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411 412[19] M.A. Vieira, L.C.C. Oliveira, R.A. Gonalves, V. Souza, R.C, Energy Fuel 23 (2009) 41359425946. 414 415[20] D.P. Torres, V.L.A. Frescura, A.J. Curtius, Microchem. J. 93 (2009) 206210. 416 417[21] M.A. Vieira, A.S. Ribeiro, A.J. Curtius, R.E. Sturgeon, Anal. Bioanal. Chem. 388 418(2007) 837847. 419 420[22] J.L. Rodrigues, D.P. Torres, V.C.O. Souza, B.L. Batista, S.S. Souza, A.J. Curtius, F. 421Barbosa Jr, J. Anal. At. Spectrom. 24 (2009) 14141420. 422 423[23] B.L. Batista, J.L. Rodrigues, J.A. Nunes, L. Tormen, A.J. Curtius, F. Barbosa Jr., 424Talanta 76 (2008) 575579. 425 426[24] J.B.B. Silva, M.A.M Silva, A.J Curtius, B. Welz, J. Anal. At. Spectrom. 14 (1999) 427 428 429[25] M. Ghisi, A.S. Ribeiro, M.A.Vieira, A.J. Curtius, Rev. Analytica 28 (2007) 58-65. 430 431[26] J.A. Nbrega; M.C. Santos; R.A. Sousa, S. Cadore, R.M. Barnes, M. Tatro, 432Spectrochim. Acta Part B 61 (2006) 465495. 433 37 38
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434[27] M. Ghisi, E.S. Chaves, D. P.C. Quadros, E. P. Marques, A. J. Curtius, A.L.B. 435Marques, Microchem. J. 98 (2011) 6265. 436 437[28] B.L. Batista, D. Grotto, J. L. Rodrigues, V.C.O. Souza, F. Barbosa Jr, Anal. Chim. 438Acta 646 (2009) 2329. 439 440 [29] L.A. Pereira, C.C. Windmller, J.B.B. Silva, Quim. Nova 34 (2011) 1167-1172. 441 442[30] A.S. Ribeiro, A.L. Moretto, M.A.Z. Arruda, S. Cadore, Mikrochim. Acta 141 (2003) 443149155. 444 445[31] J.L. Rodrigues, D.P. Torres, V.C.O. Souza, B.L. Batista, S.S. Souza, A.J. Curtius, F. 446Barbosa Jr., J. Anal. At. Spectrom. 24 (2009) 14141420. 447 448[32] F.R. Amorim , M. B. Franco, C. C. Nascentes, J. B. B. Silva, Food Anal. Meth. 4 449(2011) 4148. 450 451 452[33] R. Dobrowolski, A. Adamczyk, M. Otto, Talanta 82 (2010) 13251331. 453 454[34] A. Baysal, S. Akman, Spectrochim. Acta Part B 65 (2010) 340344. 455

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456[35] I.C.F. Damin, M.B. Dessuy, T. S. Castilhos, M. M. Silva, M.G.R. Vale, B. Welz, D.A. 457Katskov, Spectrochim. Acta Part B 64 (2009) 530536. 458 459[36] L. A. Pereira, I. Amorim, J. B.B. Silva, Talanta 68 (2006) 771775. 460 461[37] US FDA, United States Food and Drug Authorities, 2002a. Title 21- Food and Drugs. 462Chapter I - Food and Drug Administration, Department of Health and Human Services. Part 46374 - Listing of Color Additives Subject to Certification/Office of Cosmetics and Colors. 464Sec. 74.1306 D&C Red No. 6. Available from:http://www.cfsan.fda.gov/lrd/cf741306.html. 465 466[38] M. Ahamed, M, K.J. Siddiqui, Clin. Nutr. 26 (2007) 400408. 467 468 469 470 471 472 473 474 475 476 477Figure 1. Pareto chart obtained from factorial design. 478Figure 2. Response surface obtained from CCD. 41 42
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Figures Captions

479Figure 3. Peak profile of the niobium permanent modifier (a) and niobium permanent 480modifier with paladium chemical modifiers (b) in lipstick sample. 481 Figure 4. Pyrolysis and atomization curves. 482Figure 5. Lead concentration in different lipstick samples. 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 Figure 1

43 44

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499 500 501 Figure 2

502 503 504 45 46 Figure 3

23

505 506 507 508 509 510 511 512 513 Figure 4
(a) (b)

514 515

47 48

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516

Figure 5

517

49 50

25

518 519

Tables

520Table 1 . Temperature program used in optimization method. Temperature Step Drying Drying Drying Pyrolysis Atomization Clean Cool 521 522 523 524 525 526 527Table 2. Evaluation of modifiers through measurement of manufacturer's conditions. 51 52
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Ramp time (s) 5 10 10 10 1 1 1

Hold time (s) 10 20 20 10 5 10

Ar flow rate (mL/min) 250 250 250 250 0 (read) 250 250

(C) 90 150 200 700 1800 2600 20

Permanent modifier Without modifier Zr permanent, 500 g Rh permanent, 500 g Ru permanent, 500 g Nb permanent, 500 g Ta permanent, 500 g Ir permanent, 500 g

Integrated absorbance 0.183 0.163 0.193 0.146 0.202 0.152 0.139

RSD (%) 3.28 2.75 2.09 5.12 1.59 6.03 6.56

Background absorption 0.091 0.068 0.041 0.102 0.035 0.095 0.082

528* Manufactures conditions: 700 0C (PT) and 1800 0C (AT) 529 530 531 532 533 534 535Table 3. Analytical parameters of merit for for the determination of Pb in lipstick samples . Parameters Linear range (g L-1) Characteristic mass (pg) (recommended value 9.0 pg) Limit of detection, LOD Limit of quantification, LOQ Aqueous calibration curve Matrix-matched calibration curve Precision, RSD (%) 53 54 Values or range of values found 0.62 50.0 8.50 0.18 g L-1 0.62 g L-1 y = 0.0068x + 0.0196 y = 0.0076x - 0.0435 2.01 4.72
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Recovery (2.0 to 20.0 g L-1) ( %) 536 537 538 539 540 541 542 543 544

94.1 - 109

545Table 4. Mean and standard deviation values (n = 3) for Pb in of lipstick samples obtained 546by alkaline solubilization with tetramethylammonium hydroxide and microwave-assisted 547acid digestion. 548 Samples Alkaline solubilization Conc. (g g-1) Beige colors Rose colors Red color Brown colors 549 550 551 2.07 0.03 3.40 0.09 4.20 0.07 3.72 0.06 Digestion procedure Conc. (g g-1) 2.15 0.08 3.25 0.10 4.32 0.20 3.56 0.15

55 56

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