Journal of Pharmaceutical and Biomedical Analysis 55 (2011) 458–465 Contents lists available at ScienceDirect Journal of Pharmaceutical and Biomedical Analysis journal homepage: www.elsevier.com/locate/jpba Fast and fully automated analytical method for the screening of residues of aziridine and 2-chloroethylamine in pharmaceutical active principles Julián Zapata, Jorge Temprado, Laura Mateo-Vivaracho, Vicente Ferreira ∗ Laboratory for Flavor Analysis and Enology, Institute of Engineering of Aragón, I3A, Department of Analytical Chemistry, Faculty of Sciences, University of Zaragoza, 50009 Zaragoza, Spain article info Article history: Received 5 November 2010 Received in revised form 17 February 2011 Accepted 18 February 2011 Available online 24 February 2011 Keywords: Aziridine 2-Chloroethylamine SPME In fiber derivatization PFBCl abstract A simple, fast and fully automated method for the screening of aziridine (AZD) and 2-chloroethylamine (CEA) in active pharmaceutical ingredients (API) has been developed. The method is based on the in-fiber derivatization of the amines extracted from the sample headspace (previously dissolved or suspended in alkaline water) with 2,3,4,5,6-pentafluorobenzoyl chloride (PFBCl) previously adsorbed in the PDMS/DVB solid phase microextraction (SPME) fiber. The derivatives formed are further desorbed and analyzed in a gas chromatograph with negative ion chemical ionization mass spectrometry (GC–NCI-MS) using methane as reagent gas. The different operational parameters of the procedure have been optimized to get highest sensitivity. The validation of the method, however, revealed a poor repeatability, particularly evident in water-soluble APIs (RSD > 20% for AZD). In spite of that, the low detection limits (1–3 ng g -1 for AZD and CEA), speed (44 min total analysis time) and automation make that this method can be satisfactorily used as screening tool to accept or reject API batches attending to their volatile amine content and a critical specified value derived from the 1.5 g/day Threshold of Toxicological Concern (TTC) and maxima daily dosages. This was shown by analyzing seventy-five fluvoxamine maleate samples containing known levels of AZD and CEA (between 0.05 and 1.05 gg -1 ) in intermediate reproducibility conditions to get reliable estimations of precision and linearity. From these data, acceptance, rejection and non-conclusive areas of response are defined for both analytes at different confidence and replication levels using normal statistics. The method was satisfactorily applied to real fluvoxamine maleate samples. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Some volatile amines are used in the synthesis of active phar- maceutical ingredients (API) and hence may be present in small or trace amounts in the final products. Some other volatile amines are formed upon degradation of API or of some of the reagents used during the synthesis. Many of those volatile amines are dan- gerous compounds because of their toxicity and because they are also potential precursors for N-nitrosamines which are pow- erful carcinogenic agents [1–7]. This is for instance the case of 2-chloroethylamine, often used in the synthesis of many API containing amine functional groups. Residual amounts of 2- chloroethylamine can react and cycle to produce aziridine (AZD), a powerful carcinogenic agent [6,8–10]. Because of this, the synthesis and development of drugs requires a strict control of the potential volatile amines remaining in the API, and hence, there is a need for adequate analytical methods for the quantitative determination of ∗ Corresponding author. Tel.: +34 976762067; fax: +34 976761292. E-mail address: vferre@unizar.es (V. Ferreira). these compounds in API samples at gg -1 [11] level. This is the sensitivity level that would be required in most cases to ensure that, in the absence of a well defined threshold of genotoxicity, the 1.5 g/person/day TTC level is not reached [12]. Because of their volatility and the low levels at which they must be analyzed, the a priori preferred technique for the analytical determination of volatile amines should be gas chromatography [13,14]. However, the direct gas chromatographic analysis of these compounds is quite difficult because of their high reactivity, polar- ity, hydrogen-bonding character, alkaline character, and high water solubility [1,4]. Although there are some commercial chromato- graphic columns proposed for the GC-separation of underivatized amines and there are some works reporting on its use [1], their use for trace amine determination is not straightforward. It is because of this that most of the methods used are based on the chemical derivatization of the amines. The derivatization reaction seeks to decrease polarity, improve volatility, reduce chemisorption-related problems and also improve the detectability of the molecules. The derivatization process most often requires multi-step methodolo- gies and is time-consuming. A review about the reactions used for amine analysis has been presented [13]. One of the most widely 0731-7085/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.jpba.2011.02.024