Molecularly Imprinted Polymers for Dispersive (Micro)Solid Phase Extraction: A Review

The review describes the development of batch solid phase extraction procedures based on dispersive (micro)solid phase extraction with molecularly imprinted polymers (MIPs) and magnetic MIPs (MMIPs). Advantages and disadvantages of the various MIPs for dispersive solid phase extraction and dispersive (micro)solid phase extraction are discussed. In addition, an effort has also been made to condense the information regarding MMIPs since there are a great variety of supports (magnetite and magnetite composites with carbon nanotubes, graphene oxide, or organic metal framework) and magnetite surface functionalization mechanisms for enhancing MIP synthesis, including reversible addition-fragmentation chain-transfer (RAFT) polymerization. Finally, drawbacks and future prospects for improving molecularly imprinted (micro)solid phase extraction (MIMSPE) are also appraisedThis research was funded by SecretaríaXeral de Investigación e Desenvolvemento—Xunta de Galicia Grupos de Referencia Competitiva (project number ED431C2018/19), and Development of a Strategic Grouping in Materials—AEMAT (grant ED431E2018/08)S

Levels and Sources of Atmospheric Particle-Bound Mercury in Atmospheric Particulate Matter (PM₁₀) at Several Sites of an Atlantic Coastal European Region

[Abstract] Atmospheric particle-bound mercury (PHg) quantification, at a pg m⁻³ level, has been assessed in particulate matter samples (PM₁₀) at several sites (industrial, urban and sub-urban sites) of Atlantic coastal European region during 13 months by using a direct thermo-desorption method. Analytical method validation was assessed using 1648a and ERM CZ120 reference materials. The limits of detection and quantification were 0.25 pg m⁻³ and 0.43 pg m⁻³, respectively. Repeatability of the method was generally below 12.6%. PHg concentrations varied between 1.5–30.8, 1.5–75.3 and 2.27–33.7 pg m⁻³ at urban, sub-urban and industrial sites, respectively. PHg concentration varied from 7.2 pg m⁻³ (urban site) to 16.3 pg m⁻³ (suburban site) during winter season, while PHg concentrations varied from 9.9 pg m⁻³ (urban site) to 19.3 pg m⁻³ (suburban site) during the summer. Other trace elements, major ions, black carbon (BC) and UV-absorbing particulate matter (UV PM) was also assessed at several sites. Average concentrations for trace metals (Al, As, Bi, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Sb, Si, Sr, V and Zn) ranged from 0.08 ng m⁻³ (Bi) at suburban site to 1.11 µg m⁻³ (Fe) at industrial site. Average concentrations for major ions (including Na⁺, K⁺, Ca²⁺, NH₄⁺, Mg²⁺, Cl⁻, NO₃⁻ and SO₄²⁻) ranged from 200 ng m⁻³ (K⁺) to 5332 ng m⁻³ (SO₄²⁻) at urban site, 166 ng m⁻³ (Mg²⁺) to 4425 ng m⁻³ (SO₄²⁻) at suburban site and 592 ng m⁻³ (K⁺) to 5853 ng m⁻³ (Cl⁻) at industrial site. Results of univariate analysis and principal component analysis (PCA) suggested crustal, marine and anthropogenic sources of PHg in PM₁₀ at several sites studied. Toxicity prediction of PHg, by using hazard quotient, suggested no non-carcinogenic risk for adults.This work was supported by Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas ref: ED431C 2017/28-2017-2020) FEDER-MINECO (UNLC15-DE-3097, financed together (80/20%) with Xunta de Galicia and Ministerio de Ciencia, Innovación y Universidades (Programa Estatal de I+D+i Orientada a los Retos de la Sociedad ref: RTI2018-101116-B-I00 (MCIU/AEI/FEDER, UE). We are grateful to Alicia Cantarero-Roldán (SAI-University of A Coruña) for ICP-MS technical support. M. Fernández-Amado appreciates the Ministerio de Ciencia, Innovación y Universidades support (PTA2017-13607-I). The authors would like to thank P. Esperón (PTA2013-8375-I) for her support.Xunta de Galicia; ED431C 2017/28-2017-202

Exploiting dynamic reaction cell technology for removal of spectral interferences in the assessment of Ag, Cu, Ti, and Zn by inductively coupled plasma mass spectrometry

Analytical methods based on dynamic-reaction cell (DRC) technology using ammonia as a reaction gas have been developed for the determination of ultra-trace Ti, Zn, Cu and Ag by inductively coupled plasma mass spectrometry (ICP-MS). Challenging spectral interferences from complex matrices were demonstrated to be overcome by DRC, and several DRC approaches (on-mass and mass-shift) using ammonium (NH3) as a reaction gas were assessed and compared to the standard or “vented” mode analysis. Ammonium cluster ions were generated for Ti, Cu, Zn, and Ag (mass shift approach). The on-mass approach was also explored to take advantage of collisional focusing phenomena. In addition, DRC operating conditions were optimised by modifying NH3 gas flow rate and rejection parameter q (RPq). The optimised conditions were applied to show the usefulness of either on-mass or mass-shift approaches when removing Ca and P interferences. Finally, the sensitivity of all measurement modes was studied and excellent limits of detection (at few ng L−1 levels) were assessedThe authors wish to acknowledge the financial support of the Ministerio de Economía y Competitividad, Gobierno de España (project INNOVANANO, reference RT2018-099222-B-100), and the Xunta de Galicia (Grupo de Referencia Competitiva, grant number ED431C2018/19)S

Single-particle inductively coupled plasma mass spectrometry using ammonia reaction gas as a reliable and free-interference determination of metallic nanoparticles

Intensive production of nanomaterials, especially metallic nanoparticles (MNPs), and their release into the environment pose several risks for humans and ecosystem health. Consequently, high-efficiency analytical methodologies are required for control and characterization of these emerging pollutants. Single-particle inductively coupled plasma – mass spectrometry (SP-ICP-MS) is a promising technique which allows the determination and characterization of MNPs. However, several elements or isotopes are hampered by spectral interferences, and dynamic-reaction cell (DRC) technology is becoming a useful tool for free interference determination by ICP-MS. DRC-based SP-ICP-MS methods using ammonia as a reaction gas (either on-mass approach or mass-shift approaches) have been developed for determining titanium dioxide nanoparticles (TiO2 NPs), copper oxide nanoparticles (CuO NPs), copper nanoparticles (Cu NPs), and zinc oxide nanoparticles (ZnO NPs). The effects of parameters such as ammonia flow rate and dwell time on the peak width (NP transient signal in SP-ICP-MS) were comprehensively studied. Influence of NP size and nature were also investigateThe authors wish to acknowledge the financial support of the Ministerio de Economía y Competitividad (INNOVANANO projects, reference RT2018-099222-B-100), and the Xunta de Galicia (Grupo de Referencia Competitiva, grant number ED431C2018/19)S

Evolution of Gaseous and Particulate Pollutants in the Air: What Changed after Five Lockdown Weeks at a Southwest Atlantic European Region (Northwest of Spain) Due to the SARS-CoV-2 Pandemic?

[Abstract] Due to the exponential growth of the SARS-CoV-2 pandemic in Spain (2020), the Spanish Government adopted lockdown measures as mitigating strategies to reduce the spread of the pandemic from 14 March. In this paper, we report the results of the change in air quality at two Atlantic Coastal European cities (Northwest Spain) during five lockdown weeks. The temporal evolution of gaseous (nitrogen oxides, comprising NOₓ, NO, and NO₂; sulfur dioxide, SO₂; carbon monoxide, CO; and ozone, O₃) and particulate matter (PM₁₀; PM₂․₅; and equivalent black carbon, eBC) pollutants were recorded before (7 February to 13 March 2020) and during the first five lockdown weeks (14 March to 20 April 2020) at seven air quality monitoring stations (urban background, traffic, and industrial) in the cities of A Coruña and Vigo. The influences of the backward trajectories and meteorological parameters on air pollutant concentrations were considered during the studied period. The temporal trends indicate that the concentrations of almost all species steadily decreased during the lockdown period with statistical significance, with respect to the pre-lockdown period. In this context, great reductions were observed for pollutants related mainly to fossil fuel combustion, road traffic, and shipping emissions (−38 to −78% for NO, −22 to −69% for NO₂, −26 to −75% for NOₓ, −3 to −77% for SO₂, −21% for CO, −25 to −49% for PM₁₀, −10 to −38% for PM₂․₅, and −29 to −51% for eBC). Conversely, O₃ concentrations increased from +5 to +16%. Finally, pollutant concentration data for 14 March to 20 April of 2020 were compared with those of the previous two years. The results show that the overall air pollutants levels were higher during 2018–2019 than during the lockdown period.This work was supported by Ministerio de Ciencia, Innovación y Universidades (MCIU), Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) (Programa Estatal de I+D+i Orientada a los Retos de la Sociedad, ref: RTI 2018-101116-B-I00), Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas ref: ED431C 2017/28-2017-2020) and FEDER-MINECO (UNLC15-DE-3097, financed together (80/20%) with Xunta de Galicia). Joel Sánchez-Piñero acknowledges the Xunta de Galicia and the European Union (European Social Fund-ESF) for a predoctoral grant (ED481A-2018/164). María Fernández-Amado acknowledges the Ministerio de Ciencia, Innovación y Universidades (PTA2017-13607-I)

Polycyclic Aromatic Hydrocarbons Analysis in Tea Infusions and Tea Beverages Using Membrane Assisted Solvent Extraction

Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG[Abstract] Membrane Assisted Solvent Extraction (MASE) was applied as an extraction and enrichment technique of polycyclic aromatic hydrocarbons (PAHs) from tea infusions and fruit/herbal-tea beverages. PAHs have been separated and detected by high performance liquid chromatography coupled to a fluorescence detector (HPLC-FLD). Variables affecting MASE comprising extraction temperature and time, stirring rate, acceptor solvent (hexane) volume, organic modifier in the donor phase (methanol) volume, aqueous donor phase pH and ionic strength were simultaneously studied by applying a Plackett–Burman design (PBD) as screening method. Results showed statistical significance for acceptor solvent volume, extraction time and stirring rate, which were optimised by an orthogonal 23 + star central composite design (CCD). Quantitative recoveries for all PAHs (within 78–116%) were obtained by using the optimized extraction conditions: 350 µL of hexane, extraction time of 70 min and stirring rate of 175 rpm. Extraction temperature, ionic strength and donor phase pH were statistically non-significant, which simplify the procedure. The MASE method has been found sensitive (LOQs < 43 ng L−1) and precise (RSDs of < 13%). Finally, the method has been applied to assess PAHs levels in several tea infusions and fruit/herbal-tea beverages in the presence of surrogate standards. The total mean Σ16PAHs in tea infusions were from 1.2 ng L−1 (white tea) to 151.7 ng L−1 (black tea), while total mean Σ16PAHs was lower than 11.5 ng L−1 regarding tea fruit/herbal beverages. Furthermore, benzo(a)pyrene (BaP) concentrations were from < 1.5 ng L−1 (white tea) to 4.6 ng L−1 (green tea). Nevertheless, BaP concentrations obtained as well as the summation of BaA, Chry, BbF and BaP concentrations (4.6 ng L−1 for tea beverages to 7.5 ng L−1 for green tea infusions) did not exceeded the maximum levels according with European Union (EU) standards. Finally, BaP carcinogenic equivalent concentration (BaPeq) and BaP mutagenic equivalent concentration (BaPMeq) were estimated, ranging from 0.01 ng L−1 to 19.8 ng L−1 and 0.23 ng L−1 to 6.9 ng L−1, respectively.This work was supported by Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas ref: ED431C 2017/28-2017-2020) and FEDER-MINECO (UNLC15-DE-3097, financed together (80/20%) with Xunta de Galicia. Joel Sánchez-Piñero acknowledges the Xunta de Galicia and the European Union (European Social Fund - ESF) for a predoctoral grant (ED481A-2018/164)Xunta de Galicia; ED431C 2017/28-2017-2020Xunta de Galicia; ED481A-2018/16

Multi-Class Organic Pollutants in Atmospheric Particulate Matter (PM2.5) From a Southwestern Europe Industrial Area: Levels, Sources and Human Health Risk

Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG[Abstract] The occurrence of 50 multi-class pollutants comprising 18 polycyclic aromatic hydrocarbons (PAHs), 12 phthalate esters (PAEs), 12 organophosphorus flame retardants (OPFRs), 6 synthetic musk compounds (SMCs) and 2 bisphenols was studied in atmospheric particulate matter (PM2.5) samples collected at an industrial area focused on automotive manufacturing located at the Southwestern Atlantic European region (Vigo city, Spain) during 1-year period. Among all quantitated pollutants in PM2.5 samples, bisphenol A (BPA) was the most predominant with an average concentration of 6180 pg m−3, followed by PAHs comprising benzo(b+j)fluoranthene (BbF + BjF) and benzo(g,h,i)perylene (BghiP), accounting for 546 pg m−3 and 413 pg m−3 respectively. In addition, two OPFRs concerning tris(chloropropyl) phosphate (TCPP) and triphenyl phosphine oxide (TPPO) were the next following the concentration order, accounting for 411 pg m−3 and 367 pg m−3 respectively; being butyl benzyl phthalate (BBP) the most profuse PAE (56.1 pg m−3 by average). High relative standard deviations (RSDs) were observed during the whole sampling period, while statistically significant differences were only observed for PAHs concentrations during cold and warm seasons. Furthermore, some water-soluble ions and metal(oid)s were analysed in PM2.5 samples to be used as PM source tracers, whose concentrations were quite below the target levels set in the current legislation. Data obtained from principal component analysis (PCA) and PAHs molecular indices suggested a pyrogenic and petrogenic origin for PAHs, whereas occurrence of the remaining compounds seems to be attributed to resources used in the automotive industrial activity settled in the sampling area. Moreover, although a substantial anthropogenic source to PM2.5 in the area was observed, marine and soil resuspension contributions were also accounted. Finally, carcinogenic and non-carcinogenic risks posed by PM2.5-bound pollutants inhalation were assessed, being both averages within the safe level considering the whole period.This work was supported by the Ministerio de Ciencia, Innovación y Universidades (MCIU), the Agencia Estatal de Investigación (AEI) and the European Regional Development Fund (ERDF) (Programa Estatal de I + D + i Orientada a los Retos de la Sociedad, ref: RTI 2018-101116-B-I00), the Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas, refs: ED431C 2017/28 (2017–2020) and ED431C 2021/56 (2021–2024)) and ERDF-Ministerio de Economía y Empresa (MINECO) (UNLC15-DE-3097, financed together (80/20%) with the Xunta de Galicia). Joel Sánchez-Piñero acknowledges the Xunta de Galicia and the European Union (European Social Fund - ESF) for a predoctoral grant (ED481A-2018/164). Natalia Novo-Quiza acknowledges the Ministerio de Ciencia e Innovación and the European Union (ESF) for a predoctoral grant (PRE2019-088744). The Laboratorio de Medio Ambiente de Galicia (LMAG) of the Subdirección Xeral de Meteoroloxía e Cambio Climático (Xunta de Galicia) is also acknowledged for providing the samples used in the present research work. Funding for open access charge: Universidade da Coruña/CISUGXunta de Galicia; ED431C 2017/28Xunta de Galicia; ED431C 2021/56Xunta de Galicia; ED481A-2018/16

Inhalation Bioaccessibility of Multi-Class Organic Pollutants Associated to Atmospheric PM2.5: Correlation with PM2.5 Properties and Health Risk Assessment

[Abstract] Inhalation exposure to fine particulate matter (PM2.5) represents a global concern due to the adverse effects in human health. In the last years, scientific community has been adopted the assessment of the PM2.5-bound pollutant fraction that could be released (bioaccessible fraction) in simulated lung fluids (SLFs) to achieve a better understanding of PM risk assessment and toxicological studies. Thus, bioaccessibility of 49 organic pollutants, including 18 polycyclic aromatic hydrocarbons (PAHs), 12 phthalate esters (PAEs), 11 organophosphorus flame retardants (OPFRs), 6 synthetic musk compounds (SMCs) and 2 bisphenols in PM2.5 samples was evaluated. The proposed method consists of a physiologically based extraction test (PBET) by using artificial lysosomal fluid (ALF) to obtain bioaccessible fractions, followed by a vortex-assisted liquid-liquid microextraction (VALLME) and a final analysis by programmed temperature vaporization-gas chromatography-tandem mass spectrometry (PTV-GC-MS/MS). The highest inhalation bioaccessibility ratio was found for bisphenol A (BPA) with an average of 83%, followed by OPFRs, PAEs and PAHs (with average bioaccessibilities of 68%, 41% and 34%, respectively). Correlations between PM2.5 composition (major ions, trace metals, equivalent black carbon (eBC) and UV-absorbing particulate matter (UVPM)) and bioaccessibility ratios were also assessed. Principal Component Analysis (PCA) suggested that PAHs, PAES and OPFRs bioaccessibility ratios could be positively correlated with PM2.5 carbonaceous content. Furthermore, both inverse and positive correlations on PAHs, PAEs and OPFRs bioaccessibilites could be accounted for some major ions and metal (oid)s associated to PM2.5, whereas no correlations comprising considered PM2.5 major ions and metal (oid)s contents and BPA bioaccessibility was observed. In addition, health risk assessment of target PM2.5-associated PAHs via inhalation was assessed in the study area considering both total and bioaccessible concentrations, being averaged human health risks within the safe carcinogenic and non-carcinogenic levels.This work was supported by the Ministerio de Ciencia, Innovación y Universidades (MCIU), the Agencia Estatal de Investigación (AEI) and the Fondo Europeo de Desarrollo Regional (FEDER)501100008530 (Programa Estatal de I + D + i Orientada a los Retos de la Sociedad, ref: RTI 2018-101116-B-I00), the Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas, refs: ED431C 2017/28 (2017–2020) and ED431C 2021/56 (2021–2024)) and FEDER501100008530-MINECO (UNLC15-DE-3097, financed together (80/20%) with the Xunta de Galicia). Joel Sánchez-Piñero acknowledges the Xunta de Galicia and the European Union (European Social Fund - ESF) for a predoctoral grant (ED481A-2018/164). Natalia Novo-Quiza acknowledges the Ministerio de Ciencia e Innovación and the European Union (European Social Fund - ESF) for a predoctoral grant (PRE 2019-088744). The Laboratorio de Medio Ambiente de Galicia (LMAG) of the Subdirección Xeral de Meteoroloxía e Cambio Climático (Xunta de Galicia) is also acknowledged for providing the samples used in the present research workXunta de Galicia; ED431C 2017/28Xunta de Galicia; ED431C 2021/56Xunta de Galicia; ED481A-2018/16

In-Vitro Inhalation Bioavailability Estimation of Metal(Oid)S in Atmospheric Particulate Matter (PM2.5) Using Simulated Alveolar Lysosomal Fluid: A Dialyzability Approach

Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG[Abstract] A novel in-vitro method, by using synthetic body fluids, human physiological conditions and a simulated air-blood barrier (by using a dialysis membrane) has been developed and applied to assess in-vitro inhalation bioavailability of metal(oid)s associated to particulate matter (PM2,5) samples collected from an industrial site of the Northwest of Spain. A validated analytical methodology based on inductively coupled plasma mass spectrometry (ICP-MS) was used to analyse metal(oid)s concentrations in bioavailable fractions. This approach would be a more realistic human health risk assessment since considering processes that occur in human body in contrast the overestimation derived from current models (which consider environmental concentrations). Metal(oid)s such as Cu and Mo seemed to be the most bioavailable (mean in-vitro bioavailability ratios higher than 70%); Ba, Cd, Mn, Pb, Rb, Sb, Sn, V and Zn shown mean ratios between 20 and 60%, while low in-vitro bioavailability ratios (less than 20%) were observed for metal(oid)s such as Al, Co, Cr, Fe, Ni, Ti, and Tl. Health risk assessment via inhalation based on hazard carcinogenic and non-carcinogenic indexes (HIc and HInc, respectively) were performed considering three exposure scenarios using both inhalation bioavailable and total metal(oid)s concentrations in PM2.5 samples, suggesting no risk to human health. The influence of chemical composition on in-vitro bioavailability ratios was obtained, pointing out that inhalation ratios of Al, Ba, Cr, Cu, Fe, Ni, Pb and V seem to be affected by sea salt and/or crustal and/or biogenic and/or anthropogenic content of PM2.5.This work was supported by the Ministerio de Ciencia e Innovación (MCIN), the Agencia Estatal de Investigación (AEI) and the European Regional Development Fund (ERDF) (Proyectos de Generación de Conocimiento, en el marco del Programa Estatal para Impulsar la Investigación Científico-Técnica y su Transferencia, del Plan Estatal de Investigación Científica, Técnica y de Innovación 2021–2023, ref: PID2021-125201OB-I00); the Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas, refs: ED431C 2021/56 (2021–2024) and ERDF-Ministerio de Economía y Empresa (MINECO) (UNLC15-DE-3097, financed together (80/20%) with the Xunta de Galicia). Natalia Novo-Quiza acknowledges the MCIN and the European Union (European Social Fund, ESF) for a predoctoral grant (PRE2019-088744). Joel Sánchez-Piñero acknowledges the Xunta de Galicia (Consellería de Cultura, Educación e Ordenación Universitaria) and the European Union (ESF) for a predoctoral grant (ED481A-2018/164) and the Xunta de Galicia (Consellería de Cultura, Educación e Universidade) for a posdoctoral grant (ED481B-2022-002). The Laboratorio de Medio Ambiente de Galicia (LMAG) of the Subdirección Xeral de Meteoroloxía e Cambio Climático (Xunta de Galicia) is also acknowledged for providing the samples used in the present research work. Funding for open access charge: Universidade da Coruña/CISUGXunta de Galicia; ED431C 2021/56Xunta de Galicia; ED481A-2018/164Xunta de Galicia; ED481B-2022-00

Oral Bioavailability Reveals an Overestimation of the Toxicity of Polycyclic Aromatic Hydrocarbons in Atmospheric Particulate Matter

Financiado para publicación en acceso aberto: Universidade da Coruña/CISUG[Abstract] Polycyclic aromatic hydrocarbons (PAHs) in atmospheric particulate matter have adverse effects on human health, yet total PAH concentrations should overestimate the toxicity compared to the bioavailable amount of PAHs. To explore this hypothesis, we measured PAHs oral bioavailability in vitro in particulate matter with aerodynamic diameter lower than 10 µm (PM₁₀) using a test that mimics the human digestive system. This assay combines the use of simulated gastrointestinal fluids and a dialysis membrane to simulate intestinal absorption. Results show that oral PAH bioavailability was below 5%, with fluorene, anthracene, acenaphthene and phenanthrene as the most bioavailable PAHs. Data suggest no carcinogenic risk of oral bioavailable PM₁₀-bound PAHs following a health risk assessment via inhalation-ingestion by using benzo(a)pyrene-equivalent carcinogenic concentration and hazard indexes. To our best knowledge, this is the first research study of in vitro oral bioavailability estimation of PM₁₀-associated PAHs.This work was supported by Ministerio de Ciencia, Innovación y Universidades (MCIU), Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) (Programa Estatal de I+D+i Orientada a los Retos de la Sociedad, ref: RTI 2018-101116-B-I00) and Xunta de Galicia (Programa de Consolidación y Estructuración de Unidades de Investigación Competitivas ref: ED431C 2017/28-2017-2020). Joel Sánchez-Piñero acknowledges the Xunta de Galicia and the European Union (European Social Fund—ESF) for a predoctoral grant (ED481A-2018/164). The council of A Coruña is acknowledged for its assistance (collaboration agreement between the City of A Coruña and the University Institute of Environment (IUMA) of the University of A Coruña (UDC) for the measurement of PM10 particle levels in the area of Os Castros, A Coruña). The authors would like to thank EXPRELA group (UDC) for their support. Funding for open access charge: Universidade da Coruña/CISUGXunta de Galicia; ED431C 2017/28-2017-2020Xunta de Galicia; ED481A-2018/16