68 TESTING


■ Lack of somite formation ■ Lack of detachment of the tail bud from the yolk sac ■ Lack of heartbeat A criticism of the FET test has been a lack of


quantification which adequately addresses the metabolic profile of fish embryos.10


There is also


concern that embryos which are not free-feeding are, by definition, in a more protected state than a free-feeding organism would be owing to the vitelline membrane which somewhat protects the embryo from the external environment. Another limitation of this test is the volume of media. Upscale of the study design to allow accurate quantification of achieved concentration, and indeed metabolite production, would be technically challenging and may even increase the number of embryos used for testing. The FGC test has all of the restrictions that


an in vitro test intrinsically has. It is not a whole organism, and this test works on a discrete cell line derived from one organ (the gills). However, this test comprises three endpoints which do offer insight into modes of toxicity which can prove far more valuable in a weight of evidence approach. Furthermore, if support for a read- across for chronic studies is required, the FGC test offers greater insight into probable modes of action. Metabolic activity, the integrity of the cell membrane and the integrity of the lysosomal membrane are quantified in this test, and though not formally part of the validated study, it should be possible to identify key metabolites within culture media if the size of the culture allows sufficient sample volume and analytical methods exist. Undertaking the FGC in preference to the


FET not only reduces the number of organisms used, it also generates far more data of interest which can be used to bolster weight-of- evidence and read-across approaches. The study protocol is dynamic and, in theory, can be adapted for application on multiple cell lines such as epidermal, liver and the holy grail of cell culture, neurones. As understanding of key parameter analysis grows, these can also be applied to the FGC. The possibility of using cell line testing to predict neurotoxicity or even endocrine disruption potential in the environment is very real but only if the FGC approach itself is given the same support as in vitro skin sensitising testing was given.


From legacy to the future The cosmetics industry did not lead the way in the development of the environmental assessment pathway. It walks a path well- trodden by the agrochemical industry, closely followed by the biocide industry, and maybe lacks confidence in a discipline which seemingly has a language all of its own. A lack of confidence in traditional environmental testing should not, however, stop the cosmetics industry leading the way in the journey to truly animal-free testing and nor should they wait for agrochemical-focused environmental experts to recognise the gaps in the current environmental assessment approach. The future of environmental testing for


cosmetics may be predicted by understanding the agrochemical regulatory process and by following the biocides regulatory process. By


PERSONAL CARE January 2021


Figure 5: KJ Muddiman- (Coccinella sp.) Ladybird Larvae


understanding that biocides follow agrochemicals and that cosmetics follow biocides, the cosmetics industry can stay ahead of the game and by early involvement, or through industry lead proactive consideration, can influence how regulations are applied to cosmetics. In December 2019, ECHA announced the start of the development of guidance to assess the risk to bees and other arthropod pollinators, such as ladybirds, from the use of biocides. Somewhere in the regulatory future, perhaps cosmetic impact on pollinators may also be assessed. How will cosmetic consumers react to safety testing on honeybees? How can current protocols be improved to reduce, refine and replace even invertebrates used in testing? The cosmetics industry is exemplary for


recognising market trends and consumer demands and acting upon them, it has effected legislative change before and it can do it again. By championing animal alternatives, the cosmetics industry can and should lead the way in critically assessing the use of all animals in environmental assessments.


References 1 ECHA. (2014). Clarity on interface between REACH and the Cosmetics Regulation. ECHA/NA/14/46. Retrieved from https://echa.europa.eu/view-article/-/journal_ content/title/clarity-on-interface-between- reach-and-the-cosmetics-regulation


2 European Commission. (2019). Animals used for scientific purposes- Replacement, Reduction and Refinement – the “Three Rs”. Retrieved from https://ec.europa.eu/ environment/chemicals/lab_animals/3r/ alternative_en.htm


3 European Council. (2010). DIRECTIVE 2010/63/ EU OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 22 September 2010 on the protection of animals used for scientific purposes. Official Journal of the European Union. Retrieved from https:// eur-lex.europa.eu/legal content/EN/TXT/


PDF/?uri=CELEX:32010L0063&from=EN


4 Market Research Future. (2020). Cruelty-Free Cosmetics Market Research Report - Global Forecast till 2024. Retrieved May 28, 2020, from https://www.marketresearchfuture.com/ reports/cruelty-free-cosmetics-market-3825


5 North A. (2015). Flower power! Eco-friendly products on the rise. The Grocer. Retrieved from https://www.thegrocer.co.uk/household- report-2015/flower-power-eco-friendly- products-on-the-rise/518437.article


6 OECD. (2019). TG 203 Fish, Acute Toxicity Test ing. Retrieved from https://www.oecd-ilibrary. org/docserver/9789264069961-en.pdf?ex pires=1561980123&id=id&accname=guest& checksum=DB5FD00C3120C0535D63B 28D914DB68E


PC


7 OECD. (2013). TG 236, Fish Embryo Acute Tox icity (FET) Test. Retrieved from https://www. oecd-ilibrary.org/doc server/9789264203709-en.pdf?ex pires=1561984017&id=id&accname=guest& checksum=34546ABFEBC41DAC 4C041010275F1490


8 Umwelt Bundesamt. (2018). Fish Embryo Acute Toxicity (FET) test: workshop report available. Retrieved May 28, 2020, from https://www. umweltbundesamt.de/en/fish-embryo-acute- toxicity-fet-test-workshop-report.


9 ISO 21115. (2019). Water Quality- Determination of acute toxicity of water samples and chemi cals to a fish gill cell line (RTgill-W1).


10 ECHA. (2017). Expert Workshop on the poten tial regulatory application of the Fish Embryo Acute Toxicity (FET) Test under REACH, CLP and the BPR.


11 New Ideas in Marketing. (2019). Ethically-con scious consumers are creating a demand for vegan beauty products. New Ideas in Mar keting. Retrieved May 28, 2020, from https:// yougov.co.uk/topics/resources/articles-re ports/2019/05/29/ethically-conscious-con sumers-are-creating-demand-


12 OECD. (2008). Test No. 425: Acute Oral Toxicity: Up-and-Down Procedure.


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