natgillin/translations-raw huggingface dataset:
However, test substances should be compatible with one another such that they do not interact or change their chemical identity upon spiking into fish food.
Selle arvutamiseks vees soovitatakse järgida katsemeetodi C.20 (2) 6. liites sätestatud korda.
The aim is that measured results for each substance tested together should not differ greatly from the results that would be given if individual tests had been run on each test substance.
Tuleks tähele panna, et vees täheldatava kontsentratsiooni naturaallogaritmimine on sobiv juhul, kui uuendamiste vahelisel perioodil toimub eeldatavalt kontsentratsiooni eksponentsiaalne vähenemine, näiteks poolstaatilise katserežiimi puhul.
Preliminary analytical work should establish that each substance can be recovered from a multiply-spiked food and fish tissue sample with i) high recoveries (e.g. > 85 % of nominal) and ii) the necessary sensitivity for testing.
Läbivoolusüsteemi puhul ei pruugi kokkupuutekontsentratsiooni naturaallogaritmimine olla vajalik.
The total dose of substances tested together should be below the combined concentration that might cause toxic effects (cf. paragraph 51).
Kui leitakse ajaga kaalutud keskmine kontsentratsioon vees, tuleks esitada see katseprotokollis ja kasutada seda järgnevates arvutustes.
Furthermore, possible adverse effects in fish and the potential for interactive effects (e.g. metabolic effects) associated with testing multiple substances simultaneously should be taken into consideration in the experimental design.
Standardses kaladega tehtavas BCFi leidmise katses võib omastamist ja puhastumist kirjeldada kui kahte esimese järgu reaktsiooni kineetikat järgivat protsessi.
Simultaneous testing of ionisable substances should be avoided.
Omastamiskiirus = k1 × Cw
In terms of exposure, the method is also suitable for complex mixtures (cf. paragraph 13, although the same limitations in analysis as for any other method will apply).
[võrrand A5.2]
For a test to be valid the following conditions apply (cf. paragraph 24):
Üldine puhastumiskiirus = (k2 + kg + km + ke) × Cf
Water temperature variation is less than ± 2 °C in treatment or control groups
[võrrand A5.3]
Concentration of dissolved oxygen does not fall below 60 % of the air saturation value
Statsionaarses olekus on omastamiskiirus eeldusel, et kasvu ja metabolismi mõju on tühine (st kg ja km väärtust ei saa eristada nullist), võrdne puhastumiskiirusega ning seega tekib võrrandite A5.2 ja A5.3 kombineerimisel järgmine seos:
The concentration of the test substance in fish food before and at the end of the uptake phase is within a range of ± 20 % (based on at least three samples at both time points)
[võrrand A5.4],
A high degree of homogeneity of substance in food should be demonstrated in preliminary analytical work on the spiked diet; at least three sample concentrations for the substance taken at test start should not vary more than ± 15 % from the mean
on kontsentratsioon kalas statsionaarses olekus (mg/kg märgmassi alusel) ja
Concentrations of test substance are not detected, or are present only at typical trace levels, in un-spiked food or control fish tissues relative to treated samples
on kontsentratsioon vees statsionaarses olekus (mg/l).
Mortality or other adverse effects/disease in both control and test group fish should be ≤ 10 % at the end of the test; if the test is extended for any reason, adverse effects in both groups are ≤ 5 % per month, and ≤ 30 % cumulatively.
Suhtarvu k1/k2 näol on tegemist kineetilise BCFga (BCFK), mis peaks olema võrdne statsionaarset olekut iseloomustava BCFga (BCFSS), mis väljendab kalades ja vees täheldatavate statsionaarsele olekule vastavate kontsentratsioonide suhet, kuid kõrvalekaldeid võib esineda juhul, kui statsionaarse oleku saavutamises ei saa olla kindel või kui kineetilist BCFi on korrigeeritud kasvu suhtes.
If a laboratory has not performed the assay before or substantial changes (e.g. change of fish strain or supplier, different fish species, significant change of fish size, fish food or spiking method, etc.) have been made, it is advisable that a technical proficiency study is conducted, using a reference substance.
Kuna k1 ja k2 on konstandid, ei ole statsionaarse oleku saavutamine BCFK leidmiseks siiski vajalik.
The reference substance is primarily used to establish whether the food spiking technique is adequate to ensure maximum homogeneity and bioavailability of test substances.
Kirjeldatud esimest järku reaktsiooni võrranditest lähtuvalt on käesolevas 5. liites esitatud üldised arvutused, mida läheb vaja nii veekaudset kui ka toidukaudset kokkupuudet käsitlevate bioakumuleerumise hindamise meetodite puhul.
One example that has been used in the case of non-polar hydrophobic substances is hexachlorobenzene (HCB), but other substances with existing reliable data on uptake and biomagnification should be considered due to the hazardous property of HCB [47].
Järjestikuse analüüsi meetod k2 määramiseks (jaotis 4) on oluline toidukaudse kokkupuute meetodi puhul, kuna saadud väärtust on vaja nii imendumistõhususe kui ka BMFi arvutamiseks.
If used, basic information on the reference substance should be presented in the test report, including name, purity, CAS number, structure, toxicity data (if available) as for test substances (cf. paragraphs 3 and 22).
Üksnes toidukaudse kokkupuute meetodi puhul kasutatavaid arvutusi on üksikasjalikult käsitletud 7. liites.
DESCRIPTION OF THE METHOD
OMASTAMISETAPI KESTUSE HINDAMINE
Materials and apparatus should be used as described in the aqueous exposure method (cf. paragraph 26).
Enne katse tegemist saab k2 ja seeläbi aega, mis kulub statsionaarsele olekule vastavast sisaldusest teatava protsentuaalse osakaalu saavutamiseks, hinnata lähtuvalt empiirilisest seosest k2 ja süsteemi n-oktanool/vesi iseloomustava jaotuskoefitsiendi (KOW) vahel või k2 ning k1 ja BCFi vahel.
A flow-through or static renewal test system that provides a sufficient volume of dilution water to the test tanks should be used.
Tuleks siiski meeles pidada, et käesolevas jaotises esitatud võrrandid kehtivad üksnes juhul, kui omastamine ja puhastumine toimuvad vastavalt esimest järku reaktsiooni kineetikale.
The flow rates should be recorded.
Kui see ei ole ilmselgelt nii, soovitatakse omastamisetapi kestuse hindamise vajaduse korral kasutada biostatistiku ja/või farmakokineetika spetsialisti abi.
Test water should be used as described in the aqueous exposure method (cf. paragraphs 27-29).
Hinnangulist k2 väärtust (päeva kohta) saab arvutada mitme meetodiga.
The test medium should be characterised as described and its quality should remain constant during the test.
Näiteks võib esmalt kasutada järgmisi empiirilisi seoseid [1]:
The natural particle content and total organic carbon should be as low as possible (≤ 5 mg/l particulate matter; ≤ 2 mg/l total organic carbon) before test start.
(r2 = 0,95) [(3); võrrand A5.5]
TOC need only be measured before the test as part of the test water characterisation (cf. paragraph 53).
[võrrand A5.6],
Diet
kus k1 = 520 × W– 0,32 (ainete puhul, mille KOW > 3)
A commercially available fish food (floating and/or slow sinking pelletised diet) that is characterised in terms of at least protein and fat content is recommended.
(r2 = 0,85) [(4); võrrand A5.7]
The food should have a uniform pellet size to increase the efficiency of the feed exposure, i.e. the fish will eat more of the food instead of eating the larger pieces and missing the smaller ones.
jaBCF
The pellets should be appropriately sized for the size of the fish at the start of the test (e.g. pellet diameters roughly 0,6-0,85 mm for fish between 3 and 7 cm total length, and 0,85-1,2 mm for fish between 6 and 12 cm total length may be used).
(r2 = 0,90) [(5); võrrand A5.8];
Pellet size may be adjusted depending on fish growth at the start of the depuration phase.
W on uuritava ainega kokku puutuvate kalade keskmine mass (märgmassi grammides) omastamisetapi lõpus / puhastumisetapi alguses [2].
An example of a suitable food composition, as commercially supplied, is given in Appendix 7.
Muid seoseid on kirjeldatud viites 6.
Test diets with total lipid content between 15 and 20 % (w/w) have commonly been used in the development of this method.
Konstandi k2 hindamiseks võib olla kasulik rakendada keerukamat mudelit juhul, kui on näiteks tõenäoline, et aine metaboliseerumine on märkimisväärne (7, 8).
Fish food with such a high lipid concentration may not be available in some regions.
Mudeli keerukuse suurenedes tuleks aga olla saadud hinnangute tõlgendamisel ettevaatlikum.
In such cases studies could be run with a lower lipid concentration in the food, and if necessary the feeding rate adjusted appropriately to maintain fish health (based on preliminary testing).
Näiteks võib nitrorühmade esinemine viidata kiirele metaboliseerumisele, kuid see ei ole alati nii.
The total lipid content of the test group and control group diets needs to be measured and recorded before the start of the test and at the end of the uptake phase.
Seepärast peaks hindamismeetodi kasutaja katse ajakava koostamisel kaaluma saadud tulemusi keemilise struktuuri ja kõikide muude asjakohaste andmete (näiteks eelkatsete tulemuste) valguses.
Details provided by the commercial feed supplier of analysis for nutrients, moisture, fibre and ash, and if possible minerals and pesticide residues (e.g. “standard” priority pollutants), should be presented in the study report.
Statsionaarsele olekule vastavast sisaldusest teatava protsentuaalse osakaalu saavutamiseks kuluva aja võib k2 hinnangulise väärtuse põhjal leida omastamise ja puhastumise kineetikat (esimest järku reaktsiooni) kirjeldava üldvõrrandi abil, eeldusel, et kasvu ja metaboliseerumise määr on tühine.
When spiking the food with test substance, all possible efforts should be made to ensure homogeneity throughout the test food.
Kui katse jooksul täheldatakse märkimisväärset kasvu, ei ole allpool kirjeldatud hindamismeetodid usaldusväärsed.
The concentration of test substance in the food for the test group should be selected taking into account the sensitivity of the analytical technique, the test substance's toxicity (NOEC if known) and relevant physicochemical data.
Sellisel juhul on parem kasutada kasvu suhtes korrigeeritud konstanti k2g, nagu on kirjeldatud mujal (vt käesoleva liite jaotis 7).
If used, the reference substance should preferably be incorporated at a concentration around 10 % of that of the test substance (or in any case as low as is practicable), subject to analysis sensitivity (e.g. for hexachlorobenzene a concentration in the food of 1-100 μg/g has been found to be acceptable; cf. (47) for more information on assimilation efficiencies of HCB).
[võrrand A5.9]
The test substance can be spiked to the fish food in several ways depending on its physical characteristics and solubility (see Appendix 7 for more details on spiking methods):
või püsiva Cw korral
If the substance is soluble and stable in triglycerides, the substance should be dissolved in a small amount of fish oil or edible vegetable oil before mixing with fish food.
[võrrand A5.10]
In this instance, care should be taken to avoid producing a ration that is too high in lipid, taking into account the natural lipid content of the spiked feed, by adding the minimum known quantity of oil required to achieve distribution and homogeneity of the test substance in the food, or;
Statsionaarsele olekule lähenemisel (t → ∞) võib võrrandi A5.10 taandada (vt viited 9 ja 10) kujule
The food should be spiked using a suitable organic solvent, so long as homogeneity and bioavailability are not compromised (it is possible that (micro)crystals of the test substance may form in the food as a consequence of solvent evaporation and there is no easy way to prove this has not occurred; cf.
[võrrand A5.11]
(49)), or;
[võrrand A5.12].
Non-viscous liquids should be added directly to fish food but they should be well mixed to promote homogeneity and facilitate good assimilation.
Sellest lähtuvalt väljendab BCF × Cw ligikaudselt aine sisaldust kalas statsionaarses olekus (Cf-SS).
The technique for mixing should ensure homogeneity of the spiked feed.
k 2
In few cases, e.g. less hydrophobic test substances more likely to desorb from the food, it may be necessary to coat prepared food pellets with a small quantity of corn/fish oil (see paragraph 142).
Võrrandist A5.14 saab leida statsionaarsele olekule vastavast sisaldusest teatava protsentuaalse osakaalu saavutamiseks kuluva hinnangulise aja, kui võrrandite A5.5 ja A5.6 põhjal on eelnevalt arvutatud k2 hinnanguline väärtus.
In such cases, control food should be treated similarly and the final prepared feed used for lipid measurement.
Omastamisetapi statistiliselt optimaalne kestus statistiliselt vastuvõetavate andmete (BCFK) saamiseks vastab üldiselt ajavahemikule, mille jooksul kõver, mis väljendab kalas sisalduva uuritava aine kontsentratsiooni logaritmi muutust lineaarsel skaalal esitatud ajas, saavutab väärtuse, mis on vähemalt 50 % statsionaarsele olekule vastavast sisaldusest (st 0,69/k2), kuid mitte rohkem kui 95 % sellisest sisaldusest (st 3,0/k2) (11).
If used, the results of the reference substance should be comparable with literature study data carried out under similar conditions with a comparable feeding rate (cf. paragraph 45) and reference substance-specific parameters should meet the relevant criteria in paragraph 113 (3rd, 4th and 5th points).
Kui akumuleerumise määr ületab 95 % statsionaarsele olekule vastavast sisaldusest, on võimalik arvutada BCFSS.
If an oil or carrier solvent is used as a vehicle for the test substance, an equivalent amount of the same vehicle (excluding test substance) should be mixed with the control diet in order to maintain equivalency with the spiked diet.
t 95
The spiked diet should be stored under conditions that maintain stability of the test substance within the feed mix (e.g. refrigeration) and these conditions reported.
k 2
Selection of fish species
k2 = 0,652 päeva kohta
Fish species as specified for the aqueous exposure may be used (cf. paragraph 32 and Appendix 3).
Teise võimalusena võib kasutada võrrandit: teSS = 6,54 · 10 – 3 · KOW + 55,31 (tundi)
Rainbow trout (Oncorhynchus mykiss), carp (Cyprinus carpio) and fathead minnow (Pimephales promelas) have been commonly used in dietary bioaccumulation studies with organic substances before the publication of this TM.
[võrrand A5.18], et arvutada tegeliku statsionaarse oleku saavutamiseks vajalik aeg (teSS) (12).
The test species should have a feeding behaviour that results in rapid consumption of the administered food ration to ensure that any factor influencing the concentration of the test substance in food (e.g. leaching into the water and the possibility of aqueous exposure) is kept to a minimum.
Uuritava aine puhul, mille log KOW = 4, on see järgmine:
Fish within the recommended size/weight range (cf. Appendix 3) should be used.
teSS = 6,54 · 10 – 3 · 104 + 55,31 = 121 tundi.
Fish should not be so small as to hamper ease of analyses on an individual basis.
3.
Species tested during a life-stage with rapid growth can complicate data interpretation, and high growth rates can influence the calculation of assimilation efficiency [48].
PUHASTUMISETAPI KESTUSE HINDAMINE
Holding of fish
Ajavahemikku, mille jooksul aine sisaldus organismis väheneb teatava protsendini algsisaldusest, võib samuti hinnata omastamist ja puhastumist kirjeldava üldvõrrandi abil (eeldusel, et tegemist on esimest järku reaktsiooni kineetikaga; vt võrrand A5.9) (1, 13).
Acclimatisation, mortality and disease acceptance criteria are the same as for the aqueous exposure method prior to test conductance (cf. paragraphs 33-35).
Puhastumisetapi puhul eeldatakse, et Cw (toidukaudse kokkupuute katses Csööt) väärtus on null.
Pre-study work and range-finding test
kus Cf,0 on kontsentratsioon puhastumisperioodi alguses.
Pre-study analytical work is necessary to demonstrate recovery of the substance from spiked food/spiked fish tissue.
Sellest lähtuvalt arvutatakse 50 % ulatuses puhastumiseks kuluv aeg (t50) järgmiselt: Cf Cf,0 1 2 e– k2 t50 ehk t50 – ln0,50 k2 0,693 k2 Samamoodi leitakse 95 % ulatuses puhastumiseks kuluv aeg: t95 – ln0,05 k2 3,0 k2
A range-finding test to select a suitable concentration in the food is not always necessary.
Kui omastamisetapis soovitakse saavutada 80 % statsionaarsele olekule vastavast sisaldusest (kestus 1,6/k2) ja puhastumisetapis kao määr 95 % (kestus 3,0/k2), siis on puhastumisetapp umbes kaks korda pikem kui omastamisetapp.
For the purposes of showing that no adverse effects are observed and evaluating the palatability of spiked diet, sensitivity of analytical method for fish tissue and food, and selection of suitable feeding rate and sampling intervals during depuration phase etc., preliminary feeding experiments may be undertaken but are not obligatory.
Tuleb meeles pidada, et kõnealuste hinnangute puhul lähtutakse eeldusest, et omastamine ja puhastumine toimuvad vastavalt esimest järku reaktsiooni kineetikale.
A preliminary study may be valuable to estimate numbers of fish needed for sampling during the depuration phase.
Need hinnangud ei kehti juhul, kui tegemist on selgelt muu kui esimest järku reaktsiooni kineetikaga.
This can result in significant reduction in the number of fish used, especially for test substances that are particularly susceptible to metabolism.
JÄRJESTIKUSE ANALÜÜSI MEETOD: PUHASTUMISE (KAO) KIIRUSKONSTANDI K2 MÄÄRAMINE Enamiku biokontsentreerumise andmete puhul on eeldatud, et neid saab piisavalt hästi kirjeldada lihtsa kaheosalise/kaheparameetrilise mudeliga, mida väljendab sirgjoon, mis läheneb kalas puhastumisetapi ajal esineva kontsentratsiooni naturaallogaritmitud väärtustele.
Conditions of exposure
In(concentration) Cf2 Cf1 t2 t1 time
Uptake Phase duration
C0,d = ealgordinaat
An uptake phase of 7-14 days is usually sufficient, during which one group of fish are fed the control diet and another group of fish the test diet daily at a fixed ration dependent on the species tested and the experimental conditions, e.g. between 1-2 % of body weight (wet weight) in the case of rainbow trout.
[võrrand A5.21].
If needed the uptake phase may be extended based on practical experience from previous studies or knowledge of the test substance's (or analogue's) uptake/depuration in fish.
Kui on olemas ainult kahel eri ajahetkel võetud proovide andmed (näiteks minimeeritud katseplaani puhul), sisestatakse kõnealuse kahe kontsentratsiooni keskväärtused järgmisse võrrandisse:
The start of the test is defined as the time of first feeding with spiked food.
[võrrand A5.22],
An experimental day runs from the time of feeding to shortly before the time of next feeding (e.g. one hour).
kus ln(Cf1) ja ln(Cf2) on vastavalt ajahetkel t1 ja t2 mõõdetud kontsentratsiooni naturaallogaritmid ning t1 ja t2 tähistavad ajavahemikku puhastumisetapi algusest kuni kõnealuse kahe proovi võtmiseni [4].5.
Thus the first experimental day of uptake runs from the time of first feeding with spiked food and ends shortly before the second feeding with spiked food.
JÄRJESTIKUSE ANALÜÜSI MEETOD: OMASTAMISE KIIRUSKONSTANDI K1 MÄÄRAMINE (ÜKSNES VEEKAUDSE KOKKUPUUTE MEETODI PUHUL)
In practice the uptake phase ends shortly before (e.g. one hour) the first feeding with unspiked test substance as the fish will continue to digest spiked food and absorb the test substance in the intervening 24 hours.
Konstandi k1 arvutamisel järjestikuste kontsentratsiooni ajas muutumist väljendavate omastamisetapi andmete alusel kasutatakse arvutiprogrammi järgmise mudeli lähendamiseks:
It is important to ensure that a sufficiently high (non-toxic) body burden of the test substance is achieved with respect to the analytical method, so that at least an order of magnitude decline can be measured during the depuration phase.
[võrrand A5.23],
In special cases an extended uptake phase (up to 28 days) may be used with additional sampling to gain an insight into uptake kinetics.
kus k2 on teada varasematest arvutustest ning Cf(t) ja Cw(t) on vastavalt kalades ja vees ajahetkel t täheldatud kontsentratsioon.
During uptake the concentration in the fish may not reach steady-state.
Kui on olemas ainult kahel eri ajahetkel võetud proovide andmed (näiteks minimeeritud katseplaani puhul), kasutatakse k1 leidmiseks järgmist võrrandit:
Equations for estimating time to steady-state, as an indication of the likely duration needed to achieve appreciable fish concentrations, can be applied here as for the aqueous exposure test (cf. Appendix 5).
[võrrand A5.24],
In some cases it may be known that uptake of substance in the fish over 7-14 days will be insufficient for the food concentration used to reach a high enough fish concentration to analyse at least an order of magnitude decline during depuration, either due to poor analytical sensitivity or to low assimilation efficiency.
Kui selgub, et järjestikuse analüüsi meetodiga saadud hinnanguline k1 väärtus on ebausaldusväärne, tuleks k1 ja k2 arvutamiseks kasutada üheaegse analüüsi meetodit.
In such cases it may be advantageous to extend the initial feeding phase to longer than 14 days, or, especially for highly metabolisable substances, a higher dietary concentration should be considered.
Seejärel tuleks uuesti visuaalselt kontrollida saadud tõusude sobivust graafikule kantud mõõtmistulemustega.
However, care should be taken to keep the body burden during uptake below the (estimated) chronic no effect concentration (NOEC) in fish tissue (cf. paragraph 138).
Kui sobivus on endiselt väike, võib see viidata asjaolule, et tegemist ei ole esimest järku reaktsiooni kineetikaga ja tuleks kasutada keerukamat mudelit.
Depuration typically lasts for up to 28 days, beginning once the test group fish are fed pure, untreated diet after the uptake phase.
OMASTAMISE JA PUHASTUMISE (KAO) KIIRUSKONSTANTIDE ÜHEAEGSE ARVUTAMISE MEETOD (ÜKSNES VEEKAUDSE KOKKUPUUTE MEETODI PUHUL)
Depuration begins with the first feeding of “unspiked” food rather than straight after the last “spiked” food feeding as the fish will continue to digest the food and absorb the test substance in the intervening 24 hours, as noted in paragraph 126.
Konstantide k1 ja k2 väärtuse arvutamisel järjestikuste kontsentratsiooni ajas muutumist väljendavate andmete alusel võib kasutada arvutiprogramme ja järgmist mudelit:
Hence the first sample in the depuration phase is taken shortly before the second feeding with unspiked diet.
[võrrand A5.25];
This depuration period is designed to capture substances with a potential half-life of up to 14 days, which is consistent with that of bioaccumulative substances [49], so 28 days comprises two half-lives of such substances.
[võrrand A5.26],
In cases of very highly bioaccumulating substances it may be advantageous to extend the depuration phase (if indicated by preliminary testing).If a substance is depurated very slowly such that an exact half-life may not be determined in the depuration phase, the information may still be sufficient for assessment purposes to indicate a high level of bioaccumulation.
on aeg omastamisetapi lõpus.
Conversely, if a substance is depurated so fast that a reliable time zero concentration (concentration at the end of uptake/start of depuration, C0,d) and k2 cannot be derived, a conservative estimate of k2 can be made (cf. Appendix 7).
See lähenemisviis võimaldab vahetult leida hinnangulise k1 ja k2 standardvea.
If analyses of fish at earlier intervals (e.g. 7 or 14 days) show that the substance has depurated below quantification levels before the full 28-day period, then subsequent sampling may be discontinued and the test terminated.
Järjestikuse lähendamise korral soovitatakse kasutada sama andmeteisendust.Mudeli sobivust andmetega saab visuaalselt hinnata, kui kanda leitud tõusudega sirged koos proovivõtupunktide andmetega graafikule.
In few cases no measurable uptake of the test substance may have occurred at the end of the uptake period (or with the second depuration sample).
Kui sobivus on endiselt väike, võib see viidata asjaolule, et tegemist ei ole esimest järku reaktsiooni kineetikaga ja tuleks kasutada keerukamat mudelit.