ACUTE TOXICITY ESTIMATION OF MULTICOMPONENT PLANT PROTECTION PRODUCTS USING CALCULATIONS, IN SILICO AND IN VIVO METHODS. PERSPECTIVES FOR UPDATING APPROACHES TO CLASSIFICATION AND RISK ASSESSMENT

Kolesnyk S. D., Ryabukha O. S., Vasetska O. P., Bubalo N. M., Zhminko P. G., Prodanchuk M. G

BIOLOGY

Scentific article

Aim of study is to assess different alternative approaches to acute toxicity assessment of PPP, including calculation based on the assumption of additivity and in silico methods. Acute toxicity of eight PPP, containing from 2 to 5 active ingredients (AI) pesticides and number of co-formulants was assessed. In vivo studies of acute toxicity were conducted according to OECD 425. To derive in silico prediction of LD50 of active ingredients EPA Toxicity Evaluation Software Tool (T.E.S.T.) was used. Calculation of Acute Toxicity Estimate for mixture was done using GHS additivity formula, taking into account in vivo LD50 of only active ingredients of PPP, of all ingredients of formulation and LD50 of active ingredients predicted by T.E.S.T. On the basis of in vivo results, calculations and in silico predictions PPP were classified according to GHS and Ukrainian Hygienic classification. Additivity coefficients were calculated to assess extent of interaction. Results of mentioned in vivo studies, in silico modelling and calculations are presented. Mean deviation of predicted by T.E.S.T LD50 values from experimental was 62,95%. Mean deviation of calculated ATE from in vivo LD50 for mixture was 74,5% (based on in vivo LD50 data for AI only), 21,1% (based on based on in vivo LD50 data for all ingredients) and 56,4% for ATE calculated using T.E.S.T predictions. Correlation coefficients for mentioned calculations were 0,69;0,84 and 0,60 respectively. Differences of calculated and tested values of acute toxicity estimates for eight multicomponent PPP did not lead to their misclassification in up to 75% of cases according to GHS when based on in vivo data. Differences in calculated values of acute toxicity estimates based on in silico predicted results lead to misclassification of the half of the formulations, however it may be lower if account to variability of experimental results and small number of mixtures tested here. Underestimation of the hazard according to GHS classification happened only in 12,5% of the mixtures studied here. Further studies will include assessment in the similar way as presented here of larger sample of multicomponent plant protection products and other mixtures representing wider range of acute toxicity categories and development of the list of mixture type-specific LD50 values for active ingredients (e.g. depending on solvents) and their application in the PPPs classification and risk assessment.

acute toxicity, LD50, plant protection products, structure-activity relationship, classification, mixtures.

1. WHO. Chemical safety: Pesticides [Internet]. 2020. Available from: https://www.who.int/news-room/q-a-detail/chemical-safety-pesticides
2. Balan GM, Kharchenko OA, Bubalo NM. Prychyny, struktura ta klinichni syndromy hostrykh otruyen pestycydamy u pracivnykiv silskoho hospodarstva v umovakh joho reformuvannya. Suchasni problemy toksykolohiyi, harchovoyi ta himichnoyi bezpeky [Internet]. 2013;4:22- 9. Dostupno: http://protox.medved.kiev.ua/index.php/ua/issues/2013/4/item/download/428_fc18ae46392722094742062ad03b874d [in Ukrainian].
3. Ministerstvo zahystu dovkilla ta pryrodnyh resursiv Ukrainy. Derzhavniy reestr pestycidiv ta agrochimicativ, dozvolenich do vykoristnnia v Ukrainy [Internet]. 2020. Dostupno: https://mepr.gov.ua/files/docs/pesticide/2020/pesticides2008_2019+2020_32.xlsx [in Ukrainian].
4. Verchovna Rada Ukrainy. Zakon Ukrainy “Pro pestycydy ta agrochimicaty” [Internet]. Vidomosti Verchovnoyi Rady Ukrainy Verchovna Rada Ukrainy; 1995 p/91. Dostupno: https://zakon.rada.gov.ua/laws/show/86/95-вр#Text [in Ukrainian].
5. EC. Regulation (EC) No 1107/2009 of the European Parliament and of the Council of 21 October 2009 concerning the placing of plant protection products on the market and repealing Council Directives 79/117/EEC and 91/414/EEC. Off J Eur Union [Internet]. 2009;309:50. Available from: https://eur-lex.europa.eu/legal-content/EN/ALL/?uri=celex:32009R1107
6. EU. Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) [Internet]. Official Journal of the European Union 2008. p. 1-1355. Available from: https://eur-lex.europa.eu/legal-content/EN/ TXT/?uri=celex:32008R1272
7. Kolesnyk S, Bubalo N, Prodanchuk M, Zhminko P. Differences in classification for skin corrosion/irritation in EU and Ukraine: Case study of alternative (in vitro and in silico)methods application for classification of pesticide active ingredient imazamox. Toxicol Vitr. 2019;60:71-5. Available from:https://www.sciencedirect.com/science/article/abs/pii/S0887233318307835
8. Ministerstvo ohorony zdorovia Ukrainy. Gigienichna klassificacia pectycidiv za stupenem nebezpechnosti – DsanPiN 8.8.1.002-98 [Internet]. Kyiv: Ministerstvo ohorony zdorovia Ukrainy; 1998. Dostupno: https://zakon.rada.gov.ua/rada/show/va002282-98#Text [in Ukrainian].
9. Bloch D, Marx-Stoelting P, Martin S. Towards a tiered test strategy for plant protection products to address mixture toxicity by alternative approaches in human health assessment. Pest Manag Sci [Internet]. 2020 Oct 1;76(10):3326-32. Available from: https://doi.org/10.1002/ ps.6034
10. Kurth D, Wend K, Adler-Flindt S, Martin S. A comparative assessment of the CLP calculation method and in vivo testing for the classification of plant protection products. Regul Toxicol Pharmacol [Internet]. 2019;101:79-90. Available from: http://www.sciencedirect.com/science/ article/pii/S0273230018303040
11. Van Cott A, Hastings CE, Landsiedel R, Kolle S, Stinchcombe S. GHS additivity formula: can it predict the acute systemic toxicity of agrochemical formulations that contain acutely toxic ingredients? Regul Toxicol Pharmacol [Internet]. 2018;92:407-19. Available from: http://www.sciencedirect.com/science/article/pii/S0273230017304117
12. Corvaro M, Gehen S, Andrews K, Chatfield R, Arasti C, Mehta J. GHS additivity formula: A true replacement method for acute systemic toxicity testing of agrochemical formulations. Regul Toxicol Pharmacol [Internet]. 2016;82:99-110. Available from: http://www.sciencedirect. com/science/article/pii/S0273230016302963
13. Clippinger AJ, Allen D, Jarabek AM, Corvaro M, Gaça M, Gehen S, et al. Alternative approaches for acute inhalation toxicity testing to address global regulatory and non-regulatory data requirements: An international workshop report. Toxicol Vitr [Internet]. 2018;48:53-70. Available from: http://www.sciencedirect.com/science/article/pii/S0887233317303764
14. Chushak Y, Gearhart JM, Ott D. In Silico Assessment of Acute Oral Toxicity for Mixtures. Chem Res Toxicol [Internet]. 2020 Nov 18. Available from: https://doi.org/10.1021/acs.chemrestox.0c00256
15. Hamm J, Sullivan K, Clippinger AJ, Strickland J, Bell S, Bhhatarai B, et al. Alternative approaches for identifying acute systemic toxicity: Moving from research to regulatory testing. Toxicol Vitr [Internet]. 2017;41:245-59. Available from: http://www.sciencedirect.com/science/ article/pii/S0887233317300048
16. Burton J, Worth AP, Tsakovska I, Diukendjieva A. In Silico Models for Acute Systemic Toxicity. In: In Silico Methods for Predicting Drug Toxicity [Internet]. New York, NY: Humana Press; 2016. p. 177-200. Available from: http://link.springer.com/10.1007/978-1-4939-3609- 0_10
17. Bureau R. Nontest Methods to Predict Acute Toxicity: State of the Art for Applications of In Silico Methods. In: Computational Toxicology [Internet]. New York, NY: Humana Press; 2018. p. 519-34. Available from: http://link.springer.com/10.1007/978-1-4939-7899-1_24
18. Diaza RG, Manganelli S, Esposito A, Roncaglioni A, Manganaro A, Benfenati E. Comparison of in silico tools for evaluating rat oral acute toxicity. SAR QSAR Environ Res. 2015;26(1):1-27.
19. OECD. Test No. 425: Acute Oral Toxicity: Up-and-Down Procedure [Internet]. OECD; 2008 [cited 2018 Aug 29]. (OECD Guidelines for the Testing of Chemicals, Section 4). Available from: https://www.oecd-ilibrary.org/environment/test-no-425-acute-oral-toxicity-up-and-downprocedure_9789264071049-en
20. OECD. OECD Principles on Good Laboratory Practice [Internet]. Paris; 1997. (OECD Series on Principles of Good Laboratory Practice and Compliance Monitoring, Number 1). Report No.: ENV/MC/CHEM(98)17. Available from: http://www.oecd.org/officialdocuments/public displaydocumentpdf/?cote=env/mc/chem(98)17&doclanguage=en
21. EC. Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on the harmonisation of laws, regulations and administrative provisions relating to the application of the principles of good laboratory practice and the verification o. OJ L [Internet]. 2004;50:44-59. Available from: https://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX%3A32004L0010
22. Council of Europe. European convention for the protection of vertebrate animals used for experimental and other scientific purposes. Strasbourg; 1991.
23. National Research Council. Guide for the Care and Use of Laboratory Animals: Eighth Edition [Internet]. Washington, DC: The National Academies Press; 2011. Available from: https://www.nap.edu/catalog/12910/guide-for-the-care-and-use-of-laboratory-animals-eighth
24. Martin T. User’s Guide for T.E.S.T. (version 4.2). United States Environ Prot Agency [Internet]. 2013;(EPA/600/R-16/058):63. Available from: https://www.epa.gov/chemical-research/toxicity-estimation-software-tool-test
25. United Nations. Globally Harmonized System of Classification and Labelling of Chemicals (GHS) (Rev.8) (2019). 2019.
26. El-Masri HA, Reardon KF, Yang RSH. Integrated Approaches for the Analysis of Toxicologic Interactions of Chemical Mixtures. Crit Rev Toxicol [Internet]. 1997 Jan 1;27(2):175-97. Available from: https://doi.org/10.3109/10408449709021618
27. Tang H, Xu M, Zhou X, Zhang Y, Zhao L, Ye G, et al. Acute toxicity and biodistribution of different sized copper nano-particles in rats after oral administration. Mater Sci Eng C [Internet]. 2018;93:649-63. Available from: http://www.sciencedirect.com/science/article/pii/ S0928493118300869
28. Feron VJ, Groten JP, Jonker D, Cassee FR, van Bladeren PJ. Toxicology of chemical mixtures: challenges for today and the future. Toxicology [Internet]. 1995;105(2):415-27. Available from: http://www.sciencedirect.com/science/article/pii/0300483X9503239C
29. Wu X, Cobbina SJ, Mao G, Xu H, Zhang Z, Yang L. A review of toxicity and mechanisms of individual and mixtures of heavy metals in the environment. Environ Sci Pollut Res [Internet]. 2016;23(9):8244-59. Available from: https://doi.org/10.1007/s11356-016-6333-x

«Bulletin of problems biology and medicine» Issue 4 (158), 2020 year, 54-60 pages, index UDK 615.9:615.015.36:632.95: 544.165

10.29254/2077-4214-2020-4-158-54-60