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This chapter examines the mass-spectrometric instrumentation most commonly used in hair analysis, together with its evolution following both technological advancements and new analytical challenges that involve hair testing. Parallel progress of hair testing queries’ complexity and the performance of mass-spectrometric instrumentation is observed. In turn, these analytical performances are strictly connected with technological innovations and original devices concerning mass spectrometers. These two aspects are preliminary discussed in this chapter, with reference to the latest studies only. Then, an overview of the main application fields of hair testing is presented and the recent inherent literature is examined with specific consideration of the instrumental approach and procedures utilized therein. Most of these applications concern the area of forensic toxicology and in particular the detection of old and new drugs of abuse, but a relatively new topic that collects increasing interest is that of metabolomics, which is surveyed in detail.

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