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English version: SUBSTANTIATION OF FLUOPICOLIDE MAXIMUM RESIDUE LEVELS IN VEGETABLES GROWN WITH INFINITO SC 61,9 FUNGICIDE APPLICATIONS*
PeioI.M., BlagaiaA.V.
Institute of hygiene and ecology of Bohomolets National medical university, Kyiv
Fluopicoiide and Infinito 61,9 SC fungicide are pertained to the third class of hazard (moderately hazardous) by the parameters of acute toxicity via different routes of exposure of laboratory animals in accordance with the safety classification of pesticides. Fluopicoiide is not irritating the skin and mucous membranes, Infinito 61,9 SC is a light irritant of mucous membranes. Allergic properties of the compounds have not been established. Fluopicolide is a polytropic action compound. No-observed effect levels (NOEL) were set in the study of complex parameters (physiological, hematological, biochemical, histological) and were: in subchronic experiment in mice - 34.8 mg/kg, rats - 7.4 mg/kg, dogs 1000 mg/kg; in chronic experiment in mice - 7.9 mg/kg, rats - 8.4 mg/kg. The fluopicoiide acceptable dally intake (ADI) was substantiated at the level of 0.07 mg/kg of body weight, based on the lowest NOEL - 7.4 mg/kg and the safety factor 100. The study of fluopicoiide residues dynamics in vegetables grown with the appiication of Infinito 61,9 SC fungicide and taking into account toxicoiogicai properties of fluopicoiide, we substantiated the values of the maximum residue levels: in cucumbers and tomatoes - 0.04 mg/kg; in tomato juice - fluopicoiide content is not allowed. Pre-harvest intervals for cucumbers -10 days, tomatoes -14 days.
Modern agro-industry requires comprehensive protection of crops including the use of fungicides in plant chemical protection systems [1,3]. According to [2,3,4] for the cultivation of vegetables under equal conditions, excepting the use of fungicides, yield losses may account for 45-80 %.
Considering the fact that most vegetables are consumed fresh, as well as taking into account the application of chemical plant protection, fungicides should be safe for humans and for the environment [1-5]. Regarded this light we conducted sanitary and hygienic studies of Infinito 61,9 SC fungicide produced by Bayer Crop Science GmbH (Germany). Active substances of the Infinito 61,9 SC preparation are propamocarb hydrochloride (62.5 g/l) and fluopicolide (6.25 g/l). Propamocarb hydro-chloride sufficiently studied in toxicological and hygienic aspects. Propamocarb hydrochloride according to [6] refers to the third class of hazard by acute toxicity parameters via different routes of exposure. Acceptable daily intake (ADI) for humans equal to 0.02 mg/kg of body weight and required hygienic standards, in particular the maximum levels (MRLs) in cucumbers, tomatoes and tomato juice equal to 0.05 mg/kg are approved in Ukraine [5].
Fluopicolide-based preparations previously were not used in Ukraine. Hygienic regulation for fluopicolide is not performed also. Thus, Infinito 61,9 SC was tested on tomatoes and cucumbers with 1.2-1.6 l/ha (twice) application rates, and this caused the aim of our study.
Objective: hygienic evaluation of fluopicolide and propamocarb hydrochloride residues dynamics in vegetable crops grown with the use of Infinito 61,9 SC fungicide; substantiation of fluopicolide MRls in cucumbers tomatoes and tomato juice.
Objects and methods
The substance: Infinito 61,9 SC fungicide and its active ingredients fluopicolide and propamocarb hydrochlo-ride.
Active ingredient common name: propamocarb hydrochloride.
Chemical Name: Propyl -3-(dimethylamino)- propicar-bamate hydrochloride (IUPAC).
CAS registration number: 256006-41-1. Structural formula:
O
CH3
CH3
■ (CH2)3
NH3-
-C3H7
HCl
Empirical formula: CghhiClIN^C^-
Molecular weight: 224.73.
Propamocarb hydrochloride is a crystalline solid absorbent white with a specific odor, melting at 45-55°C, a density of 1.1 g/cm3. The vapor pressure is 8-10"5 Pa. Solubility in water (25°C): > 700 g/dm3. Solubility in organic solvents (g/dm3) at 25°C: methanol - > 500; methylene chloride - > 430, isopropanol - > 300; ethyl acetate - 23.0, toluene and hexane - < 0.1. Partition coefficient n-octanol/water - 2,5-10"3 [7,8].
Active ingredient common name: fluopicolide, AES638206.
Chemical name: 2,6-dichloro-N-{[3-chloro-5-(trifluoromethyl)-2- pyridinyl]methyl}benzamide (IUPAC).
CAS registration number: 239110-15-7, purity - 99.3
%.
Structural formula:
F3C
O
Cl
Empirical formula: C14H8ChF3N2O.
Molecular weight: 383.59.
Chemically pure fluopicolide is a crystalline white powder with a slight phenolic odor, melting point - 150°C, density 1.62 g/cm3 (20°C). Vapor pressure: 3,03-10"7 Pa (20°C), 8,03-10-7 Pa (25°C). Solubility in water (25°C) -2.86 mg/l (pH 1.04). Solubility in organic solvents (20°C)
O
N
C
* To cite this English version: Pelo I.M., Blagaia A.V.. Substantiation of fluopicolide maximum residue levels in vegetables grown with Infinito SC 61,9 fungicide applications/ /Problemy ekologii ta medytsyny. - 2013. - Vol 17, № 5-6. - P. 67-70.
g/m3: hexane - 0.2; ethanol - 19.2; toluene - 20.5, ethyl acetate - 37.7; acetone - 120, dimethyl sulfoxide - 180. Partition coefficient n-octanol/water - 2,9 (Pow - 79,4) [7,8].
Physical form of Infinito 61,9 SC is a liquid with a beige color and with the smell of ether, pH 7 ( 23°C), boiling point - 100°C, density 1.13 g/cm3 at 200C. Vapor pressure: 520-800 mPa at 20°C. Active substances and preparation synthesized by Bayer Crop Science GmbH (Germany). Toxicological evaluation of the preparation and its active ingredients performed according to [7,8].
Sanitary and hygienic studies are performed by us in accordance with the "Methodological guidelines for the hygienic evaluation of new pesticides" [9].
Studies of the fluopicolide and propamocarb hydrochloride content and residues were conducted in field full-scale conditions in the Kiev region, Fastiv district, Borove settl. at the Agrarian Sciences Academy Research Station during the Infinito SC application on tomatoes and cucumbers. Information on the conditions of hygienic research and fluopicolide and propamocarb hydrochloride content and residues dynamics in soil, plants and crops are presented in Tables 2, 3.
Sampling for the studies was carried out according to the "Uniform Rules of agriculture products, food and environmental objects sampling for determination of pesticides residues" [10]. Determination of fluopicolide resi-
Based on these data, Infinito 61, 9 SC and fluopi-colide according to [6] by the parameters of acute toxicity are pertained to the third class of hazard (limiting criterion - inhalation toxicity).
Fluopicolide does not irritate the skin and mucous membranes of the eyes, Infinito 61,9 SC is a weak irritant of mucous membranes according to the Draize's scale [16]. Allergenic activity of substances is not exposed in Magnusson and Kligman test.
The toxic effects of fluopicolide in subchronic (90 days) experiment were determined in mice (C57BL/6jCO strain) at 50, 200, 800 and 3200 ppm; Sprague Dawley rats at doses of 100, 1400 and 20000 ppm; Beagle dogs at doses of 5, 70 and 1000 mg/kg bw. Body weight, hematology parameters, the activity of alkaline phosphatase and cholinesterase, the content of serum albumin, cholesterol, creatinine, bilirubin, potassium and calcium in serum, and histological parameters were studied.
Taking into account the changes in these parameters the No-observed effect levels (NOEL) in subchronic experiments were set: mice - 34.8 mg/kg; rats - 7.4 mg/kg, dogs - 1,000 mg/kg.
dues was performed by gas-liquid chromatography [11,12]. The limit of detection in tomatoes, cucumbers -0.02 mg/kg soil - 0.04 mg/kg.
Determination of propamocarb hydrochloride residues has been performed by gas-liquid and high performance liquid chromatography [13,14]. The limit of detection in soil - 0.025 mg/kg, cucumbers and tomatoes - 0.025 mg/kg.
Results
Substantiation of fluopicolide MRLs in cucumbers, tomatoes and tomato juice was carried out according to the principles of integrated hygienic regulation [9,15], the main ones are:
- The daily amount of a substance that can get into the body with food, water and air shall not exceed the ADI for humans;
- Toxicological properties of the pesticide and changes in the flavor of the product must be limiting parameters for MRLs establishment [9].
The above mentioned indicates that toxicological study of active ingredient and ADI determination should be the first step in the MRL establishment.
Actually this sequence is the subject of research results presentation.
Toxic effects of the compounds are listed in Table 1.
Table 1.
Toxic effects of Infinito SC and fluopicolide.
NOELs in chronic (24 months) experiments are: mice - 7.9 mg/kg; rats - 8.4 mg/kg.
Long-term effects of fluopicolide (carcinogenic, mutagenic, embryotoxic and teratogenic effects, effects on reproductive function) are not the limiting criteria in the evaluation and substantiation of ADI to humans.
Based on the NOEL limiting magnitude - 7,4 mg/kg (rats; subchronic experiment) and applying a safety factor of 100, a substantiated value of fluopicolide ADI for humans is 0.07 mg/kg of body weight.
Fluopicolide refers to the third class of hazard (moderately dangerous) by the criterion of "persistency in soil" [6]. Tentatively allowable concentration (TAC) in soil is 0.1 mg/kg.
In line with our goal the studies of propamocarb hy-drochloride and fluopicolide residue dynamics in cucumbers and tomatoes grown with Infinito 61,9 SC application were carried out.
The results of these studies are presented in Tables 2, 3.
Criterion Animals Infinito Fluopicolide
LD50 per os rats >2500 mg/kg Wistar >5000 mg/kg n.Sprague Dawley
LD50 dermal rats >4000 mg/kg Wistar >5000 mg/kg Sprague Dawley
LC50, inhalation rats >3195 mg/m3 Wistar >5160 mg/m3 Sprague Dawley
Irritation/corrosion: -dermal -eyes New Zealand rabbits no irritation light irritation no irritation no irritation
Sensitization Guinea pigs not a sensitizer not a sensitizer
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Table 2.
Propamocarb hydrochloride and fluopicolide content in tomatoes
Day (after the last application) Content, mg/kg **
fluopicolide propamocarb hydrochloride
fruits tops fruits tops
3 0.05 ± 0.01 0.06 ± 0.01 0,12 ± 0.03 0,80 ± 0,20
7 0.03 ± 0.006 0.04 ± 0.01 0.07 ± 0.01 0,60 ± 0,15
14 0.02 ± 0.005 0.02 ± 0.005 0.02 ± 0.005 0,21 ± 0.05
20 n.d.* n.d. n.d. 0,12 ± 0.03
33 (harvest) n.d. - tomatoes n.d. - juice n.d. n.d. - tomatoes n.d. - juice 0.04 ± 0.006
Footnotes: * - n.d. - not detected,
** - propamocarb hydrochloride and fluopicolide are not detected in check samples.
Analysis of the data in Table 2 showed that fluopicolide and propamocarb hydrochloride residues in tomatoes decreased gradually. Propamocarb hydrochloride residues were not exceeding the MRL (0.05 mg/kg) on the day 14. Fluopicolide residues were 0.21 mg/kg on the day 14. Fluopicolide and propamocarb hydrochloride residues in tomatoes were not detected on the day 20. Fluopicolide and propamocarb hydrochloride residues
were not detected also at the harvest time in tomatoes and in tomato juice made from treated fruits.
We found that the organoleptic properties of tomatoes, cucumbers and tomato juice (smell, color, taste, appearance) were not affected and did not differ from control samples.
The results allow us to substantiate fluopicolide MRL in cucumbers and tomatoes at the level of 0.04 mg/kg (limit of detection of the method - 0.02 mg/kg).
Table 3.
Propamocarb hydrochloride and fluopicolide content in cucumbers and soil
Day (after the last application) Content, mg/kg **
fluopicolide propamocarb hydrochloride
fruits soil fruits soil
1 0.03 ± 0.006 0.053 ± 0.01 0,10 ± 0.03 0,62 ± 0,15
3 0.05 ± 0.01 0.05 ± 0.01 0.08 ± 0.02 0,53 ± 0,13
7 0.04 ± 0.01 0.04 ± 0.01 0.06 ± 0.01 0,20 ± 0.05
10 0.02 ± 0.005 <0.04 0.04 ± 0.006 0,10 ± 0.03
20 (harvest) n.d.* n.d. n.d. 0.06 ± 0.01
Footnotes: * - n.d. - not detected,
** - propamocarb hydrochloride and fluopicolide are not detected in check samples.
We recommend "not allowed" MRL in tomato juice as baby food (limit of detection of the method - 0.03 mg/kg). Pre-harvest intervals equal to 10 days for cucumbers and 14 days for tomatoes are substantiated.
The next stage of the study was to test the safety of our substantiated hygienic standards.
Based on the principles of integrated hygienic regulation, we have determined the total income of fluopicolide to the human body with food (tomatoes, potatoes, cucumbers, and tomato juice), water and atmospheric air, which amounted to 0.0316 mg (0.75 % of ADI).
This indicates to the public safety of vegetable products grown using Infinito 61,9 SC fungicide if the application rates and ways follow the rules (subject to the hygienic regulations, hygienic standards and pre-harvest intervals).
Conclusions
1. Fluopicolide and Infinito 61,9 SC fungicide are pertained to the third class of hazard (moderately hazardous) by the parameters of acute toxicity via different routes of exposure of laboratory animals in accordance with the safety classification of pesticides [15] .
2. Fluopicolide is not irritating the skin and mucous membranes, Infinito 61,9 SC is a light irritant of mucous membranes. Allergic properties of the compounds have not been established.
3. Fluopicolide is a polytropic action compound. No-observed effect levels (NOEL) were set in the study of complex parameters (physiological, hematological, biochemical, histological) and were: in subchronic experiment in mice - 34.8 mg/kg, rats - 7.4 mg/kg, dogs 1000 mg/kg; in chronic experiment in mice - 7.9 mg/kg, rats -8.4 mg/kg.
4. The fluopicolide acceptable daily intake (ADI) was substantiated at the level of 0.07 mg/kg of body weight, based on the lowest NOEL - 7.4 mg/kg and the safety factor 100.
5. The study of fluopicolide residues dynamics in vegetables grown with the application of Infinito 61,9 SC fungicide and taking into account toxicological properties of fluopicolide, we substantiated the values of the maximum residue levels: in cucumbers and tomatoes - 0.04 mg/kg; in tomato juice - fluopicolide content is not allowed. Pre-harvest intervals for cucumbers - 10 days, tomatoes - 14 days.
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