Monitoring of biogenic amines in organic and conventional chicken by HPLC-ESI-QTOF-MS analysis.

Monitoring of biogenic amines in organic and conventional chicken by HPLC-ESI-QTOF-MS analysis. Massimo Ricciutelli1, Franks Kamgang Nzekoue1, Giovanni Caprioli1, Manuela Cortese1, Dennis Fiorini2, Renzo Galli3, Riccardo Zeppa3, Sauro Vittori1, Gianni Sagratini1. 1 School of Pharmacy, University of Camerino, Via Sant’Agostino 1, 62032 Camerino. 2 Fileni, Localià Cerrete Collicelli, 8, 62011 Cingoli. Summary: This study presents a HPLC-QTOF-MS method to monitor 8 Biogenic amines in organic and conventional chicken breast samples. This innovative method guarantees a high sensitivity without any pre-column derivatization. Starting with similar levels at T0, BAs increment resulted lower in organic samples after 10 days of storage (T10). Keywords: HPLC-QTOF-MS, Biogenic amines, Organic chicken 1 Introduction The recent development of organic production in the poultry industry is guided by the increasing consumers interest for healthier food products. However, limited research has been performed to confirm that organic chicken (OC) products are healthier than conventional ones (CC) [1]. This could be done through the comparison of chemical quality markers levels such as Biogenic amines (BAs) between OC and CC. Indeed, BAs are a group of low-molecular-weight nitrogenous compounds present in different food matrixes. They are mainly formed through the bacterial decarboxylation of amino acids and are reported to be harmful to human health with several toxicological effects [2]. BAs determination is an analytical challenge which could be simplified and improved using high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (HPLC-QTOF-MS). The present study aims therefore to: a) assess the higher quality of OC by monitoring and comparing the levels of BAs in OC and CC samples during their shelf-life; b) to compare the BAs analysis through HPLC-QTOF-MS and HPLC-DAD. 2 Experimental 8 BAs were simultaneously determined: spermine (SPE), spermidine (SPD), cadaverine (CAD), putrescine (PUT), histamine (HIS), tyramine (TYR), 2-phenylethylamine (PHE) and tryptamine (TRY). Organic and conventional chicken breast samples were packed and stored at 4 ˚C for 2 weeks. Consecutively, BAs levels were monitored the day of production (T0) and after 10 (T10) days of storage in both chicken types. BAs were extracted with an aqueous solution of Trichloroacetic acid 5%. HPLC-QTOF-MS analyses were performed using an Agilent 1290 series instrument (Santa Clara, CA, USA), equipped with an ESI source operating in positive ionization mode with a vaporizer temperature of 350 ˚C, a nebulizer gas pressure of 55 psi; a drying gas (nitrogen) flow rate of 11 ml min−1 and a capillary voltage of 3500 V. The separation of BAs was performed using a Synergi Polar analytical column (150 × 2 mm I.D., particle size 4 µm) and a gradient mobile phase made of water (A) and acetonitrile (B) at a flow rate of 0.3 ml min−1. The injection volume was 1 µl. The mass analyzer was operating in Full Scan mode by extracting accurate mass [M+H]+ for each molecules, i.e.89.1073 m/z for PUT, 103.123 m/z for CAD, 122.0964 m/z for PHE, 138.0913 m/z for TYR, 112.0869 m/z for HIS, 161.1073 m/z for TRY, 203.225 m/z for SPE, 146.1652 m/z for SPD. 3 Results From the analyses performed on OC and CC samples, the total levels of BAs increased from T0 to T10. However, this increment was lower in OC respect to CC samples. Indeed, PHE, PUT and CAD, which were not detected at T0 in both chicken types were present at T10 but only in CC samples. TYR instead, was observed only at T10 but was higher in CC samples. On the other hand, HIS was present in both sample types at T0. However, after 10 days of storage (T10), HIS level drastically increased in CC samples while it remained constant in OC samples. HIS and TYR are considered the most toxic BAs and are predominantly relevant for food safety. SPM level was similar in both types of chicken sample and remained constant through the shelf-life. Contrarywise, SPM level decreased in CC samples and slightly increased in OC samples. The lower increment of BAs in OC samples was also observed after HPLC-DAD analyses. Nevertheless, HPLC-QTOF-MS showed the advantage that BAs were directly analysed after extraction without the need of an intermediary pre-column derivatization step as before HPLC-DAD analyses [3]. Moreover, the HPLC-QTOF-MS method showed a higher sensitivity. 4 Conclusions This study allowed to compare the levels of BAs in organic and conventional chicken during the shelf-life. From the analyses performed in HPLC-ESI-QTOF-MS, OC resulted healthier than the CC having lower levels of toxic BAs than CC after 10 days of storage. References 1. Lambio, Angel L., et al. SEARCA Professorial Chair Lecture, College, Laguna (Philippines), 21 Jun 2010 (2010). 2. Lázaro, César Aquiles, et al. LWT-Food Science and Technology. 60.1 (2015), pp 15-21. 3. Sirocchi, Veronica, et al. International journal of food sciences and nutrition 64.8 (2013): pp 921-928.