Allan
Philippe
*a,
Juraj
Košík
b,
Alexander
Welle
c,
Jean-Michel
Guigner
d,
Oliver
Clemens
e and
Gabriele E.
Schaumann
a
aGroup of Environmental and Soil Chemistry, Institute for Environmental Sciences, University of Koblenz-Landau, Fortstrasse 7, 76829, Landau, Germany. E-mail: philippe@uni-landau.de; Tel: phone: +49 6341 280 31589
bFaculty of Chemistry, Brno University of Technology, Antonínská 548/1,601 90, 75007 Brno, Czech Republic
cInstitut für Funktionelle Grenzflächen, Karlsruhe Nano Micro Facility, Karlsruhe Institute for Technology, Hermann-von-Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany
dInstitut de Minéralogie, de Physique des Matériaux et de Cosmochimie (IMPMC), Sorbonne Universities - UPMC University Paris 06, UMR CNRS 7590, MNHN, IRD UR 206, 75252 Paris cedex 05, France
eFaculty of Material Science, Technische Universität Darmstadt, Materials Design by Synthesis, Alarich-Weiss-Straße 2, 64287, Darmstadt, Germany
First published on 30th November 2017
Sunscreens are an important source of TiO2 nanoparticles in surface waters. The fate and toxicity of these particles have not been fully addressed due to the gap between model nanoparticles usually used in studies and the more complex particles found in commercial products. Therefore, mild extraction methods for TiO2 nanoparticles from sunscreens were evaluated for providing more realistic nanoparticle samples for future studies. We propose two methods based on ultrafiltration and ultracentrifugation, respectively, for extracting TiO2 nanoparticles from sunscreens using a surfactant solution as the solvent. These methods were tested on eleven commercial sunscreens with differing compositions. The ultracentrifugation variant allows extracting 250 mg from approximately 5 g of sunscreen in one day. Recoveries for ultrafiltration and ultracentrifugation were 52–96% and 78–98%, respectively. Purification efficiency was determined for the ultracentrifugation variant by determining the avobenzone concentration in sunscreen extracts using UV-spectrometry and was high for all tested sunscreens. Transmission electron microscopy and dynamic light scattering revealed a high diversity in particle shape, although size parameters were comparable (average hydrodynamic diameter: 19–34 nm). Isoelectric points were below 4.6 for all sunscreen extracts. Time-of-flight secondary ion mass spectrometry revealed that probably all TiO2 particles were coated; most of them with PDMS, some others with Al- and Si-based materials. Comparison of images of particles inside the sunscreens using cryogenic transmission electron microscopy and of extracted particles showed that while the shape of the primary nanoparticles was not affected by the extraction, they were agglomerated inside the sunscreens. These agglomerates could be completely disrupted using ultrasonication. Therefore, the particles extracted in the present study can be considered as more environmentally relevant in terms of size, shape, surface charge and coating than model TiO2 nanoparticles.
Environmental significanceSunscreens are currently the major source of TiO2 nanoparticles released into aquatic environments with partly unknown long-term effects on the concerned ecosystems. Most past experimental studies on TiO2 nanoparticles' fate or ecotoxicity focused on nanoparticles which differ strongly from the nanoparticles used in commercial products. In this study, we developed and evaluated a simple and efficient method for extracting a large amount of TiO2 nanoparticles from sunscreen products. The method was specifically optimized for providing particles for environmental studies and should contribute to render environmental studies on TiO2 nanoparticle more realistic, thus improving our ability to provide environmental risk assessment. |
In order to fill this gap, studies using particles present in commercial products for studying their fate and effects in the environment are needed. Recent estimations showed that TiO2 from sunscreen products represents 90% of the total TiO2 released into freshwater (1.6 tons per year) in Denmark.11 Therefore, particles used in sunscreens are highly relevant for environmental studies. Studies of TiO2 used in cosmetics revealed that specific aging processes (e.g. coating degradation) can be observed in environmental media.7,9 However, particle characteristics can vary strongly from one product to another. Hence, extracting realistic TiO2 particles directly from commercial products would be highly useful to improve the prediction quality of environmental studies.
Only a few extraction methods for nanoparticles from complex matrices have been reported. Ag, Au, and Pt nanoparticles were extracted from biological tissues and soil using chemical or enzymatic digestions and/or ultrasonication.12–18 However, enzymatic or acid digestion cannot be used with sunscreens as the former would be inefficient, whereas particle coating could be damaged by acids. Furthermore, separation techniques such as field flow fractionation are efficient for size measurement and quality control,19–22 but they are not practical for preparative purposes, and would require an instrument solely dedicated to continuous sample separation.23 More promising approaches involve organic solvents (chloroform, methanol, tetrahydrofuran and hexane) and in some cases ultrasonication or heating to disperse sunscreens prior to the purification of the particulate fraction.6,19,21,24 Contado et al.19 used a mixture of three solvents and ultrasound followed by a phase separation step to extract nanoparticles from one sunscreen prior to flow field flow fractionation. The extracted particles were 50–200 nm large and maximal recoveries were below 25%. Lewicka et al.6 used chloroform and centrifugation to extract nanoparticles from 8 sunscreens and characterize their size, composition and crystalline phase but did not provide recovery or surface characterization. Nischwitz and Goenaga Infante.21 compared two methods using a methanol–water mixture and/or hexane and sonication to disperse sunscreens before decantation or centrifugation of the particles. Extraction using hexane was shown to be more efficient and could recover primary particles and recoveries were 68–110%. Addition of hexane was required to stabilize the final nanoparticle suspensions. Particle sizes determined using flow field flow fractionation were between 15–40 nm. Bairi et al.24 extracted TiO2 and ZnO from eleven sunscreens using tetrahydrofuran and determined their size and crystallinity; recovery and surface characterization were not reported. In all reported studies, agglomeration after extraction was a challenge for the characterization of the particles in suspension.
However, organic solvents may alter particle coatings. For instance, polydimethylsiloxane can be dissolved in hexane, tetrahydrofuran and hexane, especially during ultrasonication.25 Furthermore, organic solvents must be removed prior to biological exposure due to their negative biological effects. This is highly important when the extracted particles should be used for ecotoxicity test or mesocosm experiments, for instance. However, the published studies on extraction techniques did not focus on the further use of the extracted TiO2 nanoparticles in environmental studies which require a large quantity of particles and a minimal alteration of the particle characteristics. Thus, a dedicated method is needed for extracting nanoparticles from sunscreens without using organic solvents, acid digestion, oxidative agents or ultrasonication.
Therefore, this study aimed at evaluating an extraction method for TiO2 nanoparticles directly from sunscreens with minimal modifications of particle characteristics and testing whether the extraction can be applied for the extraction of several grams of TiO2. The method was evaluated for its recovery and purification efficiency. In addition, all extracted nanoparticles were characterized for their size, shape, surface charge, coating, and stability towards aggregation in the extraction medium.
Number | Trade name | Type | Specification | SPF | TiO2 | ZnO | SiO2 | Al2O3 |
---|---|---|---|---|---|---|---|---|
1 | Rewe Feuchtigkeits-Sonnenspray | Lotion | For sensitive skin | 30 | Yes | No | No | No |
2 | Rewe Feuchtigkeits-Sonnencreme | Cream | For children | 50 | Yes | No | No | No |
3 | Real, Quality Sonnenmilch | Lotion | Refreshing | 30 | Yes | No | Yes | No |
4 | Real, Quality Sonnencreme | Cream | Anti-aging | 30 | Yes | No | No | No |
5 | Biotherm Lait Solaire | Lotion | — | 50 | Yes | No | No | No |
6 | Nivea Sun Pflegende Sonnenmilch | Lotion | Refreshing | 50 | Yes | No | No | No |
7 | Sundance Sonnenmilch | Lotion | Antiradical | 50 | Yes | No | Yes | No |
8 | Garnier Ambre Solaire Resisto Sonnenschutz-Milch | Lotion | For children | 50 | Yes | No | Yes | No |
9 | Alverde Sonnencreme Jojoba | Cream | For sensitive skin | 30 | Yes | No | No | Yes |
10 | Babylove Sonnencreme | Cream | For infants | 50 | Yes | Yes | No | No |
11 | Baby Sebamed Sonnenschutzlotion | Lotion | For infants | 50 | Yes | No | Yes | No |
As S8–11 were not completely dispersed in the surfactant solution, a more lipophilic solvent – n-hexane (Rotisolv HPLC, Carl Roth, Germany) – had to be used instead of an aqueous Triton X-100 solution for the first dispersion step. The sunscreen suspended in n-hexane was centrifuged in glass tubes at 5000 r.p.m. for 20 min. The n-hexane supernatant was removed using a Pasteur pipette and the remaining n-hexane was evaporated under a fume hood. The residue was redispersed in a 0.1% Triton X-100 solution (pH = 12), sonicated for 15 min, transferred to an ultrafiltration unit and centrifuged at 4500 r.p.m. for 30 min. Two further ultrafiltration steps were carried out as for S1–7. The sonication step is optional and is done to accelerate the dispersion step only.
For isoelectric point and time-of-flight secondary ion mass spectrometry (ToF-SIMS) measurements, three additional ultrafiltration and redispersion steps were performed using pure water instead of the Triton X-100 solution. This additional purification was required to reduce the pH of the solution and the surfactant concentration since a high initial pH would have required the addition of a high amount of acid for the titration during isoelectric point determination, whereas the presence of a surfactant results in a high background in ToF-SIMS.
S1, S2, S5, and S6 were chosen as a representative set of sunscreens for testing an extraction procedure on a larger scale. Using ultracentrifugation instead of ultrafiltration allowed separating larger volumes at once. 0.5 g of sunscreen and 200 mL of the 0.1% Triton X-100 solution with pH = 12 were stirred and homogenized as previously described. The suspension was transferred to one 250 mL ultracentrifuge tube made of PTFE, bath sonicated for 15 min in an ultrasonic cleaner (VWR, USA) and centrifuged at 20000 r.p.m. for 30 min using a WX Ultra Series Centrifuge from Thermo Scientific, Germany. The supernatant was carefully removed by using a Pasteur pipette and the solid residue was redispersed using 200 mL of a Triton X-100 solution. The ultracentrifugation step was repeated three times in total. For S8 and S11, the same procedure was followed for larger scale extraction with n-hexane for the first extraction step instead of the Triton X-100 solution. All extraction samples were done in triplicates.
An X-Series 2 system (Thermo, Germany) was used for ICP-MS measurements. The system was equipped with a quadrupole mass spectrometer, a platinum sample cone, a PTFE spray chamber thermostatted with a Peltier cooler and an autosampler equipped with a FAST system (ESI, Germany). The isotopes 46Ti and 47Ti were monitored as strong interferences from the diluted digestion media were observed with other isotopes. A rhodium solution (Peak Performance, California, USA) was used as an internal standard. Calibration was carried out using TiO2 (P-25, Degussa, Germany) particles, which were digested following the same procedure as that used for the samples. The recovery of the method was determined using standard addition in S5 and was 95%. No significant matrix effect could be observed from the sunscreen (see the ESI† for more details).
For the sunscreen extracts: 5 mL of the sunscreen extracts was ultracentrifuged for 35 minutes at 50000 rpm in order to remove the particulate fraction. Calculations based on technical data provided by the ultracentrifuge manufacturer showed that spherical 5 nm TiO2 nanoparticles would sediment from the top to the bottom of the tube in 27 min under those conditions. 2 mL of the supernatant was mixed with 1 mL of acetonitrile (>99.9%, HPLC grade, Sigma Aldrich, Germany) and transferred to a quartz cuvette for UV-absorbance measurements.
For the first supernatant: sunscreens were suspended as described above. S1–7 were centrifuged at 50000 r.p.m. for 30 minutes and 0.1 mL of the supernatant was diluted in 5 mL of a 1:2 acetonitrile–Triton X-100 extraction solution mixture and transferred to a quartz cuvette for UV-absorbance measurements. S8–11 were centrifuged in glass tubes at 5000 r.p.m. for 20 minutes. 0.1 mL of the supernatant was evaporated and diluted in 5 mL of the 1:2 acetonitrile–Triton X-100 mixture and transferred to a quartz cuvette for UV-absorbance measurements.
UV-absorbance was measured using a Specord50 spectrometer (Analytik Jena, Germany) at the wavelength of 355 nm (absorbance peak of avobenzone) and with an integration time of 2 s. UV measurements were repeated five times. Calibrants were prepared in the same eluent as the nanoparticle suspension using pure avobenzone (Fluka, pharmaceutical secondary standard, Germany).
• Static SIMS analysis to determine the surface compositions: the primary ion beam was rastered across a 500 × 500 μm2 field of view on the sample, and 128 × 128 data points were recorded. Primary ion doses were kept below 1011 ions per cm2 (static SIMS limit). Spectra were calibrated on the omnipresent C−, C2−, C3−, or on the C+, CH+, CH2+, and CH3+ peaks. Based on these datasets, the chemical assignments for characteristic fragments were determined.
• Surface erosion/depth profiling with an argon cluster beam to reduce surface contaminations and organic layers on the inorganic nanoparticles. Hereto, a dual beam analysis was performed in non-interlaced mode: the primary ion source was again operated in “high current bunched” mode with a scanned area of 200 × 200 μm2 (4 frames with 64 × 64 data points) and a sputter gun (operated with Ar1200+ ions, 2.5 keV, scanned over a concentric field of 400 × 400 μm2, target current 0.9 nA) was applied to erode the sample for 4 scans (6 s) followed by a 0.5 s pause to reduce surface charging from the sputter process. Argon cluster ions are eroding the softer organic layers faster compared to the harder mineral particles underneath. The total sputter time was set to 500 s corresponding to a sputter dose of 1.75 × 1015 ions cm−2. Presented spectra are integrated over time.
As concentrated surfactant solutions can damage ultrafiltration membranes (information from the supplier), lower Triton X-100 concentrations were tested. A concentration of 0.1% was chosen as it was harmless for the ultrafiltration membrane and could completely disperse S1–7 (Fig. S1†). In addition, neutral (without acid or base addition), acidic (pH = 2, HCl), and basic (pH = 12, NaOH) Triton X-100 solutions were tested. The basic solution was the most efficient dispersant for sunscreen and extracted particles based on visual inspection (Fig. S1†). Most probably, hydroxide ions can induce the partial hydrolysis of ester groups present in major components of several sunscreens (e.g. octocrylene, alkyl benzoates, 2-ethylhexyl salicylate) resulting in more hydrophilic products and, therefore, in more efficient dispersion of the sunscreen. Therefore, a basic solution of 0.1% Triton X-100 was used to disperse and purify the sunscreens tested in this study.
Ultrafiltration is advantageous for extracting lower amounts (15 mL per tube in this study) of sunscreens as the cut-off is more accurate and depends mainly on the geometrical size of the molecules or particles to be retained, whereas ultracentrifugation separates particles based on their size and density. On the other side, the cut-off of ultracentrifugation can be adapted by changing the rotation speed. In addition, this technique was more adapted for separating large amounts of dispersed sunscreen. Therefore, a larger scale separation method using 250 mL ultracentrifugation tubes was tested. This method allowed preparing 1 L of final isolated TiO2 nanoparticle suspension (approximately 250 mg L−1, based on average final concentrations obtained in this study) in one working day. This is an improvement compared to a recently reported extraction method21,24 in which tetrahydrofuran was used as a solvent, since the reported procedure took more than one day to be completed. The absence of both organic solvent in the final suspension and of ultrasonic treatments is an advantage of the method evaluated in this study compared to other previously reported methods.19,21
No | TiO2 content in sunscreens (% (w/w)) | TiO2 concentration ultrafiltration (mg L−1) | Recovery (%) | TiO2 concentration ultracentrifugation (mg L−1) | Recovery (%) |
---|---|---|---|---|---|
1 | 4.1 ± 0.3 | 231 ± 31 | 96.0 ± 7.2 | 240 ± 17 | 90.6 ± 5.3 |
2 | 6.1 ± 0.7 | 256 ± 19 | 73.2 ± 6.7 | 375 ± 31 | 94.4 ± 9.2 |
3 | 5.5 ± 0.4 | 272 ± 17 | 88.2 ± 8.6 | ||
4 | 4.0 ± 0.5 | 210 ± 20 | 83.0 ± 7.4 | ||
5 | 4.1 ± 0.2 | 209 ± 21 | 87.3 ± 6.0 | 213 ± 26 | 94.5 ± 7.6 |
6 | 5.2 ± 0.8 | 281 ± 24 | 88.2 ± 7.8 | 212 ± 12 | 78.0 ± 4.0 |
7 | 6.0 ± 0.9 | 312 ± 37 | 73.7 ± 12.3 | ||
8* | 5.5 ± 0.1 | 272 ± 10 | 95.7 ± 2.3 | 270 ± 33 | 98.0 ± 6.2 |
9* | 13.1 ± 0.7 | 342 ± 9 | 51.7 ± 5.6 | ||
10* | 5.9 ± 0.2 | 251 ± 11 | 83.5 ± 7.9 | ||
11* | 6.4 ± 0.5 | 239 ± 25 | 72.8 ± 6.3 | 308 ± 27 | 98.2 ± 5.4 |
Recoveries in terms of TiO2 particles were determined by dividing the TiO2 concentrations in extracted particle suspensions by the TiO2 concentration in the sunscreen and the values ranged between 72–98% for all methods and samples except for S9 with a value near 51% (Table 2). Considering the TiO2 content and recovery, S9 seems to be an exception. The producer claimed that this sunscreen contained mainly plant extracts and TiO2. Therefore, the matrix of this sunscreen strongly differed from the rest of the tested sunscreens. Nonetheless, recoveries are overall highly satisfying, since they are comparable to recoveries obtained by Nischwitz et al. (64–110%).21 Recoveries for the methods using ultracentrifugation and ultrafiltration were comparable.
For ultrafiltration, the final volume of the particulate fraction in the concentrate can be controlled by setting the time or the speed of the centrifugation step. Therefore, the concentration of the non-particulate compounds relative to TiO2 would be decreased by a factor of 1000–6000 after three successive filtration steps considering the final volume of the concentrates (0.5–1 mL for our samples).
A similar estimation of the purification rate could not be achieved for ultracentrifugation, since the removal of the supernatant was difficult to reproduce. Indeed, pellets were not observed for all samples tested in this study after ultracentrifugation. Thus, the volume of the removed supernatant had to be adapted for each sample. Therefore, the efficiency of the ultracentrifugation technique in terms of removal of the molecular matrix was tested in order to quantify the variation of the purification efficiency for different samples. This can be achieved by determining the concentration of the non-particulate fraction before and after purification by ultracentrifugation.
As sunscreen compositions are diverse and complex, a systematic measurement of all compounds in the suspension of extracted TiO2 would be especially tedious and inefficient. Therefore, we used 1-(4-methoxyphenyl)-3-(4-tert-butylphenyl)propane-1,3-dione (avobenzone) as a representative of the non-particulate fraction. Avobenzone was selected because it is a widely used UV-A blocker and was one of the main ingredients in all tested sunscreens except S9 and S10. In addition, avobenzone has an absorption peak at 355 nm which renders possible a selective quantification in the presence of Triton-X (absorption peak at 280 nm) using UV-spectrometry. We assumed that avobenzone was the only compound absorbing at that wavelength in our sunscreen extracts despite the possible interferences of other organic UV blockers in the sunscreens such as octocrylene (absorbance peak at 305 nm). This an acceptable working hypothesis since the determination of absolute concentration of avobenzone was not required for determining the purification rate.
The concentrations of avobenzone in the sunscreen extract were between 0.1 and 25.1 mg L−1 (Table 3) and were, therefore, much lower than the concentrations measured in the first supernatant (before the first centrifugation step). In this study, we defined the purification rate as the ratio between the TiO2/avobenzone mass to mass ratio calculated in the first supernatant and the TiO2/avobenzone ratio in the final TiO2 extracts. Purification rates ranged from approximately 8 to 2002 (Table 3). In other words, the concentration of avobenzone relative to TiO2 was divided by 8 to 2002 after purification. These values can be compared to the purification efficiency determined for S11 using the ultrafiltration method (Table 3) which was 5417 and, thus, in the range of the efficiency estimated from the remaining volume after filtration (see above).
Sunscreen no. | Avobenzone concentration in the first supernatant in mg L−1 | Avobenzone concentration in the final extract in mg L−1 | Purification rate |
---|---|---|---|
1 | 282.4 ± 0.2 | 0.16 ± 0.01 | 2002 |
2 | 165.0 ± 0.4 | 25.09 ± 0.06 | 8 |
5 | 117.1 ± 0.2 | 1.63 ± 0.01 | 76 |
6 | 145.5 ± 0.3 | 1.86 ± 0.01 | 60 |
8 | 268.3 ± 0.7 | 8.37 ± 0.01 | 32 |
11 | 338.6 ± 0.4 | 0.24 ± 0.01 | 1420 |
11* | 338.6 ± 0.4 | 0.049 ± 0.007 | 5417 |
The complexity of the matrix and the multiple possible interactions between the various dissolved compounds (e.g. sorption, macromolecular assemblies) during the centrifugation process could explain the differences observed between the sunscreen extracts. The observed variations between samples in terms of purification efficiency indicate that great care has to be taken when using ultracentrifugation for purifying nanoparticles extracted from complex mixtures. We also recommend to, at least, estimate the purification rate for applying this method to further sunscreens. It has to be noted that it is always possible to increase the purity of the nanoparticle extracts by carrying out further purification steps, if the target experiments require a high purity.
The TiO2 particles used in sunscreens were very diverse. The three main types of shapes were observed: spherical, irregular, and elongated (Fig. 1). Energy dispersive X-ray spectroscopy confirmed that the particles contained Ti (Fig. S2†). Almost all particles observed were agglomerated in sunscreens 1 and 7, whereas some isolated primary particles were observed in sunscreens 5 and 9. Nonetheless, most of the observed particles were agglomerated. Therefore, we can assume that most of the nanoparticles present in the sunscreen were agglomerated prior to extraction.
Fig. 1 Representative images of TiO2 particles in sunscreens obtained using transmission electron microscopy in cryogenic mode. The sunscreen number is given on the upper right corner. |
Interestingly, TiO2 nanoparticles were frequently observed agglomerated on large (several hundreds of nanometers) spherical particles (Fig. 1). The weak contrast compared to the water background suggests that these particles were composed of organic materials. We suppose that these spherical objects are organic components of the sunscreens in the form of emulsion in water. As the water is frozen, these lipophilic drops would be trapped in the ice matrix.27 The fact that TiO2 nanoparticles were often attached on their surfaces suggests that their coating is lipophilic. Cryo-TEM images suggest a high concentration (especially in S5 and S9) of these organic particles in the sunscreen. Therefore, we can exclude that these particles are hard polymer spheres, since they would be concentrated with the inorganic particles during ultrafiltration. In fact, this is not reflected by the TEM and DLS analyses of the extracted fractions (see below). Indeed, cryo-TEM pictures suggest a high concentration of these organic particles in the sunscreens, especially in 5 and 9 (Fig. 1), which should strongly influence the DLS results and be observable in classical TEM.
Fig. 2 Representative images of extracted inorganic nanoparticles from eleven commercial sunscreens obtained using transmission electron microscopy. The sunscreen number is written on the upper right corner. TEM pictures of the other sunscreens extract can be found in the ESI† (Fig. S3). TEM images from S2, S6, and S10 were similar to S1, whereas images from S3 and S8 were similar to S7 and S5, respectively. |
No | Average length (nm) | Average width (nm) | Particle shape | Average hydrodynamic diameter (nm) | Isoelectric point | Proposed surface coating |
---|---|---|---|---|---|---|
1 | 19.9 ± 6.7 | 14.2 ± 5.0 | Spherical, irregular | 23.2 ± 1.2 | 2.6 | PDMS |
2 | 23.4 ± 7.2 | 15.0 ± 4.8 | Spherical and angular | 28.0 ± 1.0 | 2.2 | PDMS |
3 | 35.5 ± 12.0 | 15.9 ± 3.9 | Ellipsoidal and angular | 35.5 ± 2.0 | 1.7 | PDMS |
4 | 13.4 ± 3.1 | 7.5 ± 1.7 | Spherical | 19.3 ± 0.8 | 1.9 | PDMS |
5 | 36.6 ± 11.6 | 7.3 ± 2.5 | Elongated | 24.8 ± 0.8 | 1.9 | PDMS |
6 | 24.2 ± 6.6 | 15.0 ± 4.3 | Spherical | 34.3 ± 0.6 | <1.8 | SiO2 |
7 | 32.5 ± 12.1 | 13.9 ± 3.7 | Ellipsoidal | 30.8 ± 1.0 | <1.8 | PDMS |
8 | 29.3 ± 10.0 | 9.3 ± 3.7 | Elongated, spherical and ellipsoidal | 22.3 ± 0.8 | 2.1 | PDMS |
9 | 42.0 ± 12.5 | 22.7 ± 7.5 | Spherical, angular and elongated | 35.6 ± 0.6 | 4.5 | Al2O3 |
10 | 48.8 ± 16.6 | 31.5 ± 12.6 | Spherical | 37.6 ± 1.5 | 4.4 | Al(OH)3 |
11 | 27.0 ± 11.4 | 12.4 ± 3.9 | Ellipsoidal and spherical | 27.4 ± 1.7 | 3.1 | Al2O3 + SiO2 |
The range of average primary particle sizes determined from TEM pictures was surprisingly narrow (length: 20–50 nm, width: 7–32 nm, Table 4). Furthermore, low standard deviations indicate that particles used in the sunscreens are fairly monodisperse. It was not possible to image any coating at the surface of nanoparticles using TEM or HR-TEM due to the low contrast between the coating and the carbon from the grid and the thinness of the coating layer.
The average hydrodynamic diameter obtained using DLS after sonication and dilution was measured for all extracted sunscreens (Table 4). Sonication time and dilution ratio were optimized for obtaining primary particle size and, thus, obtaining reproducible size measurements (Tables S2 and S3†). Despite particle shape differences, there is a good correspondence between the sizes of primary particles determined using TEM and the average hydrodynamic diameter obtained using DLS. This indicates that the extracted particles were in the form of primary particles or small agglomerates after extraction and sonication. Thus, extracted particles are disagglomerated during the dispersion processes. If it is required to obtain agglomerated particles, it is still possible to replace the surfactant based solution with pure water by further ultracentrifugation of ultrafiltration steps and, hence, induce re-agglomeration of the particles if the agglomerated form is crucial for the targeted investigations. However, extracting the particles from sunscreens without modifying their original agglomeration structure is still challenging as it would imply to avoid introducing any shear forces or stabilizing agent, which would not result in the dispersion of most sunscreens.
Determination of the isoelectric points is a classical approach to qualitatively estimate the surface charge behavior of colloids in aqueous media.28 Furthermore, particles are expected to agglomerate faster at pH near the isoelectric point due the lack of electrostatic repulsion.29 Therefore, we determined isoelectric points by measuring ζ-potentials at several pH values. The complete ζ-potential–pH curves can be found in the ESI† (Fig. S4), while the isoelectric points measured for each sunscreen are summarized in Table 4. The presence of the surfactant in the solution could influence the absolute ζ-potential value by shifting the shear plane on the particle surface. However, the isoelectric point is not affected as Triton X-100 is a neutral surfactant and the formation of micelles did not affect the measurements as verified with a blank sample. While S9–11 had an isoelectric point between 3 and 4.5, other extracts had an isoelectric point lower than 3. Nanoparticles extracted from sunscreens were thus all negatively charged at pH > 4 in aqueous NaCl solution. The ζ-potentials at pH = 5 were varying between 0 and −30 mV (Fig. S4†). Thus, particle stability in terms of agglomeration in aqueous media can be expected to vary strongly depending on the sunscreen used.
ToF-SIMS measurements allowed a deeper insight into the nature of the nanoparticles' coating. It has to be noted that most of the surfactant present in the extraction media was removed before ToF-SIMS measurements since its high concentration would have resulted in a thick layer of surfactant over the particles and would have disturbed the surface analysis. Fragments characteristic for typical coating materials (Al+, SiOH+, Zn+, polydimethylsilane (PDMS): SiC3H9+ at 73.05 m/z, and Si2C5H15O+ at 147.08 m/z) used in sunscreens7,9,30 and TiO+ as a marker of the bulk material were monitored before and after sputtering with Ar clusters. Several Zr+ isotope signals were monitored in addition but were insignificant for all samples measured. A decrease in the signal intensity for a given fragment during sputtering combined with an increase of the TiO+ signal intensity indicates that the observed fragment originates from the topmost surface layer, which is removed during sputtering.31 As an example, ToF-SIMS measurements for S1 (Fig. 3) are quite clear in that respect. The TiO+ signal intensity increased with sputtering, indicating an increased exposure of the bare TiO2 surface. On the other hand, the signal intensity of characteristic PDMS fragments dramatically decreased during sputtering, suggesting the absence of PDMS in the deeper layers. Therefore, we can conclude that PDMS is most probably present on the surface layer of the TiO2 nanoparticles extracted from S1. In this case, the weak SiOH+ signal more probably originates from the fragmentation of PDMS than from an additional underlying silica based coating layer as the SiOH+ signal intensity decreased after sputtering. Signal intensities of other ions were not significant (lower than for the blank sample).
Fig. 3 ToF-SIMS signal intensities obtained without (full line) and with (dashed line) Ar-clusters sputtering for S1. Vertical lines indicate the exact mass expected from the respective expected ions or fragments; from left to right: 27Al+, 28SiOH+, 48TiO+, 68Zn+, (CH3)3Si+, and (CH3)3SiOSi(CH3)2+. The latter two are characteristic fragments for PDMS.30 |
The other sunscreen extracts were analyzed following the same procedure (Fig. S5–15†). Absolute intensities for TiO+ varied strongly between samples despite efforts made in selecting a scanned area completely covered with particles. This is probably due to visible differing topologies of the particle layer due to different agglomeration and deposition behaviors during the drying process and to the particle shape diversity. However, ToF-SIMS results from S1–5, S7, and S8 had similar patterns and may have, therefore, similar surface chemistry, except for S2 and S5, for which the PDMS signals were dominant. This can be explained by the presence of a PDMS layer thicker than other samples for which the surface coating was almost completely removed after the first erosion step. The erosion of a thick PDMS layer would take more time and, therefore, the signal integrated over time can become higher than the signal obtained from the first measurement (static SIMS). Therefore, we concluded that particles extracted from S1–5, S7, and S8 were most probably all coated with PDMS although with most likely differing coating thicknesses. Estimation of the coating thickness was not possible due to the above mentioned irregularities of the surface topology. Al+ signals higher than the blank were observed for S5 and S8–11 indicating an Al-based coating such as Al2O3 or Al(OH)3; in accordance with the formulation mentioning “alumina” for S10 and S11 and “aluminum hydroxide” for S5 and S10. No Al-containing compounds were mentioned in S8 and S11. However, the Al concentration in these sunscreens may be too low to be mentioned on the packaging as suggested by the weak Al+ signals compared to TiO+. As no other significant signals were observed for S9, we conclude that the nanoparticles are coated solely with Al2O3. Zn was detected in S10 and S11. This was expected for S10 since it contains ZnO nanoparticles in addition to TiO2 nanoparticles. It has to be noted that, due to the presence of two types of nanoparticles in S10, it should remain undecided to which type the observed alumina coating belongs. The Zn signal from S11 most probably results from residual sorption of Zn2+ ions on the TiO2 nanoparticles from “zinc stearate” present in the formulation. SiOH+ was significant for S6 and S11, which is noticeable because the PDMS signals were weak for both samples. This suggests the presence of a silica based coating. Since amorphous SiO2 could be partly dissolved under alkaline conditions (pH > 10),32 it cannot be completely excluded that S9 and S11 originally contained SiO2 as a surface coating. The types of coating which are expected from the ToF-SIMS measurements are summarized in Table 4.
Since it is highly challenging to characterize surface coatings inside sunscreens due to the high organic background present in the matrix, it was impossible to quantify to which extent the surface coating is altered during the extraction procedure. Nonetheless, the fact that we could detect several coatings typical for TiO2 particles used in sunscreens indicates that the proposed extraction method does not alter the surface coating or, at least, partially preserve it. Furthermore, the characterization of the nanoparticles extracted in this study suggests some similarities in terms of size, shape and surface composition between the extracted nanoparticles and ingredients used in the cosmetic industry. For instance, S1–4, S7 and S11 are similar to the NM-103, reported in other studies, which has an average length of 22 nm and a broadness of 34 nm and a spherical to elongated shape.10,33 On the other side, S5 and S8–9 match the description of the T-lite™ SF from BASF with an elongated shape and lengths between 50–200 nm and broadness of 5–10 nm.7,9 NM-103 and T-lite™ SF are coated with PDMS as most of the sunscreen extracts in the present study. However, NM-103 had an isoelectric point of 8.2, which is much higher than the isoelectric points measured in our study and for T-lite™ SF.7,9 Therefore, we recommend using more than one single reference material for environmental studies in order to cover a broad range of nanoparticle characteristics as encountered in commercial products.
Furthermore, the proposed method can also be used as a quality control method for commercial products, especially in combination with separation techniques such hydrodynamic chromatography or flow field flow fractionation for fast particle characterization. In addition, this study provides a representative overview on which types of TiO2 nanoparticles are present in commercial sunscreens and is, therefore, informative for future risk assessment of nanoparticles in surface waters.
Footnote |
† Electronic supplementary information (ESI) available. See DOI: 10.1039/c7en00677b |
This journal is © The Royal Society of Chemistry 2018 |