Journal of Chromatography A, 1252 (2012) 115–129 Contents lists available at SciVerse ScienceDirect Journal of Chromatography A jou rn al h om epage: www.elsevier.com/locat e/chroma Direct analysis of pharmaceuticals, their metabolites and transformation products in environmental waters using on-line TurboFlow TM chromatography–liquid chromatography–tandem mass spectrometry Rebeca López-Serna a , Mira Petrovi ´ c b,c,* , Damià Barceló a,b a Department of Environmental Chemistry, IDAEA-CSIC, C/Jordi Girona 18-26, 08034 Barcelona, Spain b Catalan Institute for Water Research (ICRA), C/Emili Grahit, 101, Cientific and Technologic Park of Girona University, 17003 Girona, Spain c Catalan Institution for Research and Advanced Studies (ICREA), C/Lluís Companys, 23, 08010 Barcelona, Spain a r t i c l e i n f o Article history: Received 24 April 2012 Received in revised form 19 June 2012 Accepted 24 June 2012 Available online 1 July 2012 Keywords: Pharmaceuticals Transformation products Metabolites, TurboFlow chromatography On-line LC–MS/MS Environmental waters a b s t r a c t The work describes the development and validation of an analytical method for simultaneous determi- nation of 58 pharmaceuticals and 19 metabolites and transformation products in environmental waters. The method is fully automated and consists of a direct injection of a small volume (several mL) of water sample to an on-line system composed of TurboFlow TM chromatography for the extraction and clean-up followed by liquid chromatography–electrospray-tandem mass spectrometry (TFC-LC–ESI-MS/MS). The feasibility and limitations of the technique in the analysis of environmental and wastewaters is discussed. The main advantages include high throughput, minimum sample manipulation, low error introduction, high selectivity, sensibility and reliability. The method provided a tool for the determination of a high number of active metabolites and transformation products and was successfully applied in the analysis of samples from the river Ebro basin. © 2012 Elsevier B.V. All rights reserved. 1. Introduction Pharmaceuticals are detected in the environment in the ng L -1 –g L -1 range [1], that may not pose much acute risk, how- ever little is known about long-term effects and other receptors in non target organisms that could be sensitive [2]. Just a small portion (less than 10%) of the 3200 pharmaceuticals registered in Europe and North America have been measured in environmental samples. According to Howard and Muir [3] of 275 pharmaceuticals detected in the environment, 92 were rated as potentially bio-accumulative and 121 were rated as potentially persistent. Nevertheless, even those ones with relatively short environmental half-lives can be considered “pseudo-persistent” in the environment, due to their continual introduction into surface waters via wastewater treatment plant (WWTP) discharges [4]. Moreover, even though individual concentrations of any pharmaceutical compound might be low, the combined concentrations from compounds sharing a common mechanism of action could be substantial [2,5,6]. In addition to that, there must be taken into consideration that the compound may pass through several processes of transformation * Corresponding author at: Catalan Institute for Water Research (ICRA), C/Emili Grahit, 101, Cientific and Technologic Park of Girona University, 17003 Girona, Spain. Tel.: +34 972183380; fax: +34 972183248. E-mail address: mpetrovic@icra.cat (M. Petrovi ´ c). and degradation from the moment of intake, or even before, from its synthesis, until it arrives the environment. The term “degradation” here refers to the elimination of the parent compound without any knowledge whether the compound is mineralized or transformed in something else [7]. Products include both metabolites excreted via urine or faeces, and transformation products (TP) which can be formed by physico-chemical and biological processes in WWTPs or water works and/or in the environment, from parent pharma- ceuticals and/or their metabolites released [8]. While substantial data exists regarding the occurrence and ecotoxicology of parent pharmaceuticals, much less is known about their metabolites and TP, whose existence is even unknown in most of cases. Very few pharmaceutical metabolites and TP are studied or monitored in the environment, so there is a need to strengthen the research in this area [8]. Therefore, the development of multi-residue analytical meth- ods that will permit simultaneous monitoring of parent compounds and their metabolites and TPs is crucial to properly assess the risk posed by the presence of pharmaceuticals in the environment. This is of great importance as a synergistic effect of different pharma- ceuticals on aquatic life might occur and has to be investigated [9]. Due to the very low levels of pharmaceuticals that may be present, an analyte pre-concentration procedure is compulsory in order to obtain the desired levels of analytical sensitivity, which often requires relatively large sample volumes (100–1000 mL) to be processed [10,11]. As awareness of more sustainable and 0021-9673/$ – see front matter © 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.chroma.2012.06.078