**Chemical Characterization of Electronic Cigarette (e-cigs) Refill Liquids Prior to EU Tobacco Product Directive Adoption: Evaluation of BTEX Contamination by HS-SPME-GC-MS and Identification of Flavoring Additives by GC-MS-O**

**Jolanda Palmisani 1,\*, Carmelo Abenavoli 2, Marco Famele 2, Alessia Di Gilio 1,\*, Laura Palmieri 1, Gianluigi de Gennaro <sup>1</sup> and Rosa Draisci <sup>2</sup>**


Received: 3 February 2020; Accepted: 6 April 2020; Published: 10 April 2020

**Abstract:** The present study focused on the determination of benzene, toluene, ethylbenzene and xylenes (BTEX) concentration levels in 97 refill liquids for e-cigs selected by the Italian National Institute of Health as representative of the EU market between 2013 and 2015 prior to the implementation of the European Union (EU) Tobacco Product Directive (TPD). Most of the e-liquids investigated (85/97) were affected by BTEX contamination, with few exceptions observed (levels below the limit of quantification (LOQ) of headspace-solid phase micro extraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) methodology). Across brands, concentration levels ranged from 2.7 to 30,200.0 μg/L for benzene, from 1.9 to 447.8 μg/L for ethylbenzene, from 1.9 to 1,648.4 μg/L for toluene and from 1.7 to 574.2 μg/L for m,p,o-xylenes. The variability observed in BTEX levels is likely to be related to the variability in contamination level of both propylene glycol and glycerol and flavoring additives included. No correlation was found with nicotine content. Moreover, on a limited number of e-liquids, gas chromatography-mass spectrometry-olfactometry (GC-MS-O) analysis was performed, allowing the identification of key flavoring additives responsible of specific flavor notes. Among them, diacetyl is a flavoring additive of concern for potential toxicity when directly inhaled into human airways. The data reported are eligible to be included in the pre-TPD database and may represent a reference for the ongoing evaluation on e-liquids safety and quality under the current EU Legislation.

**Keywords:** electronic cigarettes; flavoring additives; BTEX; contamination; headspace solid micro phase extraction; gas chromatography-olfactometry; human health; EU regulation
