Appl. Phys. A 69 [Suppl.], S179S182 (1999) / Digital Object Identier (DOI) 10.

1007/s003399900277

Applied Physics A
Materials Science & Processing
Springer-Verlag 1999

Quantitative analysis of trace metal accumulation in teeth using laser-induced breakdown spectroscopy
O. Samek1, , D.C.S. Beddows2 , H.H. Telle2 , G.W. Morris2 , M. Liska1 , J. Kaiser1
1 Institute of Physical Engineering, Technical 2 Department of Physics, University of Wales

University of Brno, Technicka 2, 616 69 Brno, Czech Republic Swansea, Singleton Park, Swansea SA2 8PP, United Kingdom

Received: 21 July 1999/Accepted: 11 September 1999/Published online: 28 December 1999

Abstract. The technique of laser ablation is receiving increasing attention for applications in dentistry, specically for the treatment of teeth (e.g. drilling of micro-holes and plaque removal). In the process of ablation a luminous micro-plasma is normally generated which may be exploited for elemental analysis. Here we report on quantitative Laser-Induced Breakdown Spectroscopy (LIBS) analysis to study the presence of trace minerals in teeth. A selection of teeth of different age groups has been investigated, ranging from the rst teeth of infants, through the second teeth of children, to adults to trace the inuence of environmental factors on the accumulation of a number of elements in teeth. We found a close link between elements detected in tooth llings and toothpastes with those present in teeth. PACS: 32.30Jc; 42.62Be; 42.62Fi

Common exposure of teeth to numerous chemical compounds is encountered when eating and drinking, or when brushing teeth for cleaning. Our preliminary study suggests that metals may be absorbed or adsorbed by the tooth during the standard procedure of brushing teeth for cleaning, as well as by migration from tooth llings. Using the technique of LIBS we tried to establish whether this effect is a surface or short distance effect only, or whether migratory processes are encountered.

1 Experimental The LIBS experiments were carried out in the usual manner. Briey, short pulses of laser light are focused on the material under test. During the light-matter interaction period the sample material is heated, vaporized and atomized, thus ejecting a small amount of material from the surface. For sufciently high laser pulse energy the result is a luminous plasma. Spectral analysis of the light emitted by the plasma can yield both qualitative and quantitative information on trace elements contained in the material. A standard Nd:YAG laser (Brilliant, Quantel) was used to generate the LIBS plasma. It was running at the fundamental wavelength of 1064 nm, at a repetition rate of 20 Hz. Individual laser pulses had a pulse length of about 46 ns, and could be adjusted for pulse energies of 10100 mJ using a Glan polarizer. The laser light was focused by a lens onto the surface of the samples teeth, tooth llings, or toothpastes. Light from the plasma was collected by a single quartz lens, which focused the plasma light emission onto an optical bre bundle, used for imaging the light on the entrance slit of a spectrometer. Spectral analysis was carried out using a standard spectrograph (ACR500, Acton) with a gateable, intensied photodiode array detector (IPDA1024, Princeton Instruments) attached to it; the gating of the detector and the timing for spectral accumulation were controlled by a PC via a pulse delay generator (PG200, Princeton Instruments).

The technique of Laser-Induced Breakdown Spectroscopy (LIBS), or Laser Micro Analysis (LMA) offers a simple and fast method of elemental analysis. Any material solid, liquid, or gaseous can be analysed, and none or only very little sample preparation is needed. Detection limits for solid samples are usually in the range of a few tens of parts per million. While this may be inferior to other methods of analysis, one has to keep in mind that LIBS analysis can be undertaken directly from the solid sample, often in situ or even remotely [1, 2]. When considering the role of mineral nutrition and metabolism in the context of maintaining human health, knowledge of the presence or absence of key elements is of vital importance. The accumulation of various elements in excess concentration and these may indeed only be extremely low may have toxic effects. Such elements include for example aluminium, cadmium, mercury, and lead.
Corresponding

author. (Fax: +42-5/4114-2842, E-mail: ota@fyzika.fme.vutbr.cz) COLA99 5th International Conference on Laser Ablation, July 1923, 1999 in Gttingen, Germany

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As is usual in analytical laser spectroscopy, the setup needs optimization for best sensitivity and reproducibility. In agreement with work carried out in this laboratory and by other research groups, one usually nds that higher laser pulse energies generate hotter plasmas and hence more light emission. However, in this study we did not exceed pulse energies of about 20 mJ, since at higher energies crater formation became severe. All experimental results described below were recorded using laser pulses of energy 20 mJ, focused by a lens with f = 25 cm. The diameter of the laser spot on the sample was about 0.10.2 mm. Under these excitation conditions, atomic emission lines exhibited the best signal-to-noise ratio and reproducibility when the delay between plasma generation and collection of light emission was 15 s. This depended on the element of interest (note that strong Ca lines were observed up to 40 s delay). The observation window was about 3 s. A full description of the experimental setup and an evaluation of the various measurement parameters may be found in [3]. 2 Results and discussion Using the experimental setup outlined above a selection of teeth from different age groups, four different types of teeth llings and ve samples of toothpaste have been studied. Spectra were recorded in the range 250800 nm. Both major constituents of the sample matrix and trace elements could be identied and partially quantied, including Ag, Al, Ca, Cr, Hg, K, Mg, Mn, Na, Ni, P, Sn, Ti, and Zn. 2.1 Toothpastes and llings Teeth are commonly exposed to numerous chemical compounds, being encountered when eating or when brushing them for cleaning. For the latter, normally well-dened concentrations of different elements can be associated with specic brands; hence we have investigated samples of a few different toothpastes to identify typical elemental constituents. In Fig. 1 spectra for two different brands of toothpastes from one manufacturers range are shown, in the wavelength range 389401 nm. Relatively high contents of aluminium and titanium in addition to Ca were found in the toothpaste which contained a whitening additive (Fig. 1a). For the sample without this additive only Ca lines are visible in the same spectral range (Fig. 1b). Thus one may suspect that, when using the toothpaste which contains the whitening additive, migration of Al onto and into the tooth may be found. In dental treatment of tooth decay, llings of amalgam are frequently used which contain different mixtures of metals. Alternatively, various novel lling materials have become popular (one example of such lling materials was included in this study). Four different types of teeth lling materials were investigated to identify the presence of potentially toxic elements; these may migrate into the tooth through long exposure by its presence in it. For two different teeth lling materials, the spectral range 251261 nm is displayed in Fig. 2.

Fig. 1a,b. Segments of LIBS spectra of toothpastes a with and b without whitening additive; selected elemental lines are indicated

Fig. 2a,b. Segments of LIBS spectra for two different lling materials: a white lling material, b amalgam. Prominent spectral emission lines for some metals are indicated

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The specic range was selected because it contains strong emission lines from potentially toxic elements, namely aluminium, titanium, mercury, and tin. Differences in the lling material are quite evident, and may be used to identify specic types of lling; no attempt at quantitative elemental analysis has been made at this stage of the investigation. 2.2 Cross-section and depth proling of teeth After preliminary experiments with toothpastes and tooth lling materials a number of teeth samples were investigated. At the rst instance, we have targeted Al as an element present both in some tooth llings and toothpastes. Thus its migration into the tooth itself may be found in specimens which were exposed to the relevant sources of Al. Two different methods were used to investigate the samples. Firstly, simple depth proling of the tooth was attempted. The aim was to analyze the samples by laser drilling. This approach is somewhat limited because of the hole drilling, preventing the luminous plasma from being observed without geometrical restriction once a deep crater has been generated. Hence, only surface and near-surface features can be analyzed. Secondly, cross-sections of teeth were investigated allowing us to scan across or through the tooth. For this, the samples were cut, horizontally or vertically, into thin slices (about 1.5 mm thick). The method can be expanded to generate two-dimensional distribution maps of elements in the tooth. This will be described in a forthcoming paper.

It is worth noting that our investigation represents a spot measurement technique because the diameter of the area under investigation was only about 0.10.2 mm. An example of the cross-section method of a tooth with a lling inside is shown in Fig. 3. The spectrum displayed in Fig. 3a was recorded for the lling which was roughly circular in shape in the centre of the tooth cross-section; the spectrum shown in Fig. 3b was taken about 2 mm away from the lling. It is obvious that the migration of Al has reached a large distance from the lling, and aluminium was present throughout the tooth at different levels of concentration. In a nal series of measurements a selection of teeth of different age groups was studied, ranging from the rst teeth of infants, through the second teeth of children, to adults, using both methods noted further above. Relative concentrations of Al in all specimens (semi-quantitative data) are collected in Table 1. Note that the values for the relative Al concentrations are normalized to the Al signal obtained from the lling in tooth sample 20. As mentioned earlier, in this study we only attempted to establish the presence of Al in the samples. Traces of Al were presented in about a third of the teeth used in this preliminary study (total of 20 specimen); the spectra were similar to that shown in Fig. 3. In the same samples under investigation, Al was only found at or near the surface (depth proling method) but, for other samples, Al was found scattered inside the teeth (cross- section method). Surprisingly, Al was present within two rst teeth of children where no lling was present. It should be noted that only the trace element Al has been investigated in detail thus far because numerous lines of other elements of interest were overlapping with strong Ca lines at the setting of the spectrometer resolution for this survey study. Work on a systematic study of different elements such as Cr, Hg, Sn, Ti, and Zn is currently in progress. We would also like to note that, in addition to numerous lines from different elements which we were able to identify, a few emission lines from the specimen under investigation

Table 1. Relative concentration of Al in tooth samples. Al signals from lling of sample 20 is the reference value c = 1 Sample # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Age group Infant Infant Infant Infant Infant Child Child Child Child Child Child Adults Adults Adults Adults Adults Adults Adults Adults Adults Filling No No No No No No Yes No No No Yes Yes No No Yes No Yes Yes No Yes Relative concentration 0 0.012 0 0 0.011 0 0 0 0 0.015 0 0 0 0 0.022 0 0.020 0.017 0.013 1 = lling

second second second second second second

teeth teeth teeth teeth teeth teeth

Fig. 3a,b. Spectral emission lines of Al at two different locations in a tooth with lling: a centred on the lling and b 2 mm away from the lling

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could not be assigned yet, although we recorded spectra for most possible metal elements for comparison (pure metal samples provided by Goodfellows Ltd were used). This suggests that the matrix of biological samples behaves differently from that of a solid metal target, and it could be that some element lines which are not visible in LIBS plasmas generated from solid metal sample matrices could be quite prominent in the biological samples. For example, we observe strong emission from phosphorus (P) from the tooth samples; in contrast, P is extremely difcult to detect as an additive in steels. Further studies on this aspect are in progress.

air at standard atmospheric pressure which largely eliminated the need for any special sample preparation. For selected samples we attempted to detect a range of elements normally present in trace amounts in biological specimen. Concentrations of aluminium found in the teeth under investigation can most likely be attributed to the use of toothpaste with whitening additives and the presence of tooth llings.
Acknowledgements. This work was supported in part by Grants 101/98/ P282 and 101/97/K009 from the Czech Grant Agency.

References 3 Conclusions We have shown that the elemental analysis of teeth (which constitute non-metallic, biological matrices) is feasible using LIBS. Furthermore, we have performed the measurements in
1. M. Sabsabi, P. Cielo: Appl. Spectrosc. 49, 499 (1995) 2. C.M. Davies, H.H. Telle, D.J. Montgomery, R.E. Corbett: Spectrochim. Acta, Part B 50, 1059 (1995) 3. O. Samek, M. Liska, J. Kaiser, V. Krzyzanek, D.C.S. Beddows, A. Belenkevitch, G.W. Morris, H.H. Telle: SPIE 3570, 263 (1999)