Abstract
One of the most challenging aspects of interpreting quantitative information of biological samples from laser inductively coupled plasma mass spectrometry (LA-ICP-MS) is a lack of appropriate matrix-matched internal standards that is needed for calibrations. There are standards available; however, most certified reference standard materials are suboptimal, due to the high variability and complexity of biological materials, especially for calcified tissues. In the present study, we described an approach in which bovine bone pellets are used as reliable matrixmatched standards for quantitative analysis of bone samples. Bovine tibial bones, sourced from a local butcher shop, were treated with or without autoclave sterilization. The samples were lyophilized over a 24 hour period, after which the elemental distributions in autoclaved, non-autoclaved bone pellets and naïve bone fragments were investigated using inductively coupled plasma optical emission spectrometry (ICP-OES) and LA-ICP-MS methods, in addition to homogeneity analysis of non-autoclaved bone pellets. The results demonstrated that non-autoclaved and autoclaved bone pellets shared similar average elemental concentrations after correcting for background signal; natural bone fragments, on the other hand, showed large sample variations. Factors such as low cost and ease of manufacture, “home-made” non-autoclaved bone pellets are the preferred option and these were subjected to further investigations. The homogeneity analysis revealed that non-autoclaved bone pellets had a higher degree of homogeneity, with minimal standard deviations and a uniform particle size of less than 100 μm. These results show that non-autoclaved bovine bone pellets are reliable and easy-to-make alternative to matrix-matched reference material with which to analyse calcified tissues by LA-ICP-MS.
Highlights
The atomic distribution and concentration of beneficial or toxic metals and non-metals in biological tissues is of great interest
Standard Reference Materials (SRM) 610 and SRM 612 were purchased from the National Institute of Standards and Technology (Gaithersburg, MD, USA) and used to demonstrate accuracy and precision of the LA-ICPMS method for determining of elemental mass fractions in pelletized bovine bone samples
Signal intensities obtained during LA-ICP-MS analysis were sufficient to detect all of the selected isotopes above the gas blank for autoclaved, non-autoclaved bone pellets, naïve bone fragments and NIST SRM 610 and SRM 612 (Table 2)
Summary
The atomic distribution and concentration of beneficial or toxic metals and non-metals in biological tissues is of great interest. LA-ICP-MS can depict analytic distributions in biological tissue sections with a spatial resolution ranging from 10-100 μm [2] and offers a detection limit from parts per million to as low as parts per billion [3]. These are benefits that make LA-ICP-MS a versatile tool in mapping the elements making up soft and hard tissues, thereby contributing better understanding of biological processes especially where the quantifications of these trace elements are concerned [1,4,5]. In human tissues this method has been used with biopsy sections from breast [10], lymph node [11], brain [12] and eye lenses [13], and of particular relevance for the present study, ICP-MS analyses have been reported for mineralised tissues [14,15,16]
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