Optimization and validation of a micro-QuEChERS method for phthalates detection in small samples of cetacean blubber

Annalisa Sambolino; Marta Rodriguez; Jesus De la Fuente; Manuel Arbelo; Antonio Fernández; Manfred Kaufmann; Nereida Cordeiro; Ana Dinis

doi:10.1016/j.mex.2023.102502

Optimization and validation of a micro-QuEChERS method for phthalates detection in small samples of cetacean blubber

MethodsX. 2023 Nov 29:12:102502. doi: 10.1016/j.mex.2023.102502. eCollection 2024 Jun.

Authors

Annalisa Sambolino^{1

2

3}, Marta Rodriguez^{1

2}, Jesus De la Fuente⁴, Manuel Arbelo⁴, Antonio Fernández⁴, Manfred Kaufmann^{3

5}, Nereida Cordeiro^{2

5}, Ana Dinis¹

Affiliations

¹ MARE - Marine and Environmental Sciences Centre, ARNET - Aquatic Research Network, Regional Agency for the Development of Research, Technology and Innovation (ARDITI), Funchal, Portugal.
² LB3, Faculty of Exact Science and Engineering, University of Madeira, Funchal, Portugal.
³ Faculty of Life Sciences, University of Madeira, Funchal, Portugal.
⁴ Division of Histology and Animal Pathology, University Institute for Animal Health and Food Security (IUSA), Veterinary School, University of Las Palmas de Gran Canaria, Canary Islands, Spain.
⁵ CIIMAR-Interdisciplinary Centre of Marine and Environmental Research, 4450-208 Matosinhos, Portugal.

Abstract

In this study, an innovative method was developed to detect and quantify phthalates in fresh cetacean blubber. An adaptation of the ammonium formate QuEChERS method was used and adapted as a micro-extraction for small quantities of samples. Significantly, this technique utilized minimal quantities of reagents and salts, with the additional implementation of rigorous Quality Assurance/Quality Control protocols to further reduce background contamination. To ensure the reliability of this method, comprehensive validation procedures were conducted, with a specific focus on two widely studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus). Determination coefficients (R²) for matrix-matched calibration were >0.93 with limits of quantifications (LOQ) of the method in the range of 5-10 ng/g. Mean recovery values were between 40 and 100 %. This novel methodology holds particular relevance for environmental research studies, offering the capability to detect emerging contaminants with minimal sample requirements. This aspect is particularly valuable in investigations that involve free-ranging animals and rely on biopsy sampling. It allows for the assessment of contaminant levels in healthy individuals within wild populations, enhancing our understanding of ecological impacts and potential conservation measures.•A micro-extraction adaptation of the ammonium formate QuEChERS method was developed and applied to a small quantity of fresh cetacean blubber to detect phthalates.•Small quantities of reagents and salts were used, and additional Quality Assurance/ Quality Control procedures were taken to further minimize background contamination.•Method validation was carried out for two cosmopolitan and extensively studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus).

Keywords: Ammonium formate “Micro – QuEChERS” method for phthalates detection in small quantities of cetacean blubber; Bottlenose dolphin; GC–MS; Gas chromatography-mass spectrometry; Phthalic acid esters; Pilot whale; Plastic additives.