natgillin/translations-raw huggingface dataset:
Prior to routine use of a validated method that adheres to this Test Method, laboratories may wish to demonstrate technical proficiency, using the ten substances recommended in Table 1.
Pred rutinsko uporabo validirane metode, pri kateri se upošteva ta testna metoda, je možno, da želijo laboratoriji z uporabo desetih snovi, ki so priporočene v tabeli 1, dokazati tehnično strokovnost.
Under this Test Method, the UN GHS optional category 3 is considered as no category.
V okviru te testne metode ni neobvezne kategorije 3 GHS ZN.
For novel similar (me-too) test methods developed under this Test Method that are structurally and functionally similar to the validated reference methods or for modifications of validated methods, the performance standards described in the Appendix to this Test Method should be used to demonstrate comparable reliability and accuracy of the new test method prior to its use for regulatory testing.
Za podobne nove testne metode, ki so razvite v okviru te testne metode ter so strukturno in funkcionalno podobne validiranim referenčnim metodam, ali za spremembe validiranih metod, se morajo pred začetkom uporabe novih testnih metod kot predpisanega testiranja za dokazovanje njihove primerljive zanesljivosti in natančnosti upoštevati standardi učinkovitosti, opisani v Dodatku k tej testni metodi.
Proficiency Substances which are a subset of the Reference Substances listed in the Appendix
Napovedne snovi, ki so podmnožica referenčnih snovi, navedenih v Dodatku
In vivo score
Rezultat testa in vivo
GHS category
Kategorija GHS
naphthalene acetic acid
naftalen ocetna kislina
No Cat.
kat. št.
methyl stearate
metil stearat
heptyl butyrate
heptil butirat
hexyl salicylate
heksil salicilat
1-bromohexane
1-bromoheksan
butyl methacrylate
butil metakrilat
The following is a description of the components and procedures of a reconstructed human epidermis model test for skin irritation assessment.
V nadaljevanju so opisani elementi in postopki testa za ocenjevanje razdraženosti kože z modelom rekonstruirane človeške pokožnice.
A reconstructed human epidermis model can be constructed, prepared or obtained commercially (e.g. EpiSkin™, EpiDerm™ and SkinEthic RHE™).
Model rekonstruirane človeške pokožnice se lahko izdela, pripravi ali pridobi na tržišču (npr. EpiSkin™, EpiDerm™ in SkinEthic RHE™).
Standard test method protocols for EpiSkin™, EpiDerm™ and SkinEthic RHE™ can be obtained at [http://ecvam.jrc.ec.europa.eu](21, 22, 23).
Protokoli standardnih testnih metod za EpiSkin™, EpiDerm™ in SkinEthic RHE™ so na voljo na spletni strani [http://ecvam.jrc.ec.europa.eu](21, 22, 23).
Testing should be performed according to the following:
Testiranje se mora izvajati v skladu z naslednjimi zahtevami:
Reconstructed Human Epidermis Model Components
Elementi modela rekonstruirane človeške pokožnice
General model conditions
Splošni pogoji modela
Normal human keratinocytes should be used to construct the epithelium.
Za izdelavo epitelija se uporabijo običajni človeški keratinociti.
Multiple layers of viable epithelial cells (basal layer, stratum spinosum, stratum granulosum) should be present under a functional stratum corneum.
Pod funkcionalno roženo plastjo je več slojev živih epitelijskih celic (bazalna plast, trnasta plast, zrnata plast).
Stratum corneum should be multilayered containing the essential lipid profile to produce a functional barrier with robustness to resist rapid penetration of cytotoxic marker substances, e.g. sodium dodecyl sulphate (SDS) or Triton X-100.
Rožena plast mora biti razdeljena na več plasti in vsebovati lipidni profil za močno funkcionalno pregrado za zaustavitev hitre penetracije označevalcev citotoksičnosti, npr. natrijevega dodecil sulfata (SDS) ali Tritona X-100.
The barrier function may be assessed either by determination of the concentration at which a marker substance reduces the viability of the tissues by 50 % (IC50) after a fixed exposure time, or by determination of the exposure time required to reduce cell viability by 50 % (ET50) upon application of the marker substance at a specified, fixed concentration.
Funkcija pregrade se lahko oceni z določitvijo koncentracije, pri kateri označevalec po določenem času izpostavljenosti zmanjša viabilnost tkiv za 50 % (IC50), ali z določitvijo časa izpostavljenosti, ki je potreben za zmanjšanje viabilnosti celic za 50 % (ET50) po nanosu določene stalne koncentracije označevalca.
The containment properties of the model should prevent the passage of material around the stratum corneum to the viable tissue, which would lead to poor modelling of skin exposure.
Zadrževalne značilnosti modela morajo preprečiti prehod materiala okoli rožene plasti v živo tkivo, kar bi preprečilo učinkovitost modelov za raziskovanje v zvezi z izpostavljenostjo kože.
The skin model should be free of contamination by bacteria, viruses, mycoplasma, or fungi.
V modelu kože ne sme biti bakterij, virusov, mikoplazme ali plesni.
Functional model conditions
Pogoji za funkcionalen model
Viability
Viabilnost
The preferred assay for determining the magnitude of viability is the MTT (20).
Zaželena analiza za določanje stopnje viabilnosti celic je z MTT (20).
The optical density (OD) of the extracted (solubilised) dye from the tissue treated with the negative control (NC) should be at least 20 fold greater than the OD of the extraction solvent alone.
Optična gostota izvlečka (topnega) barvila iz negativnega kontrolnega tkiva mora biti vsaj dvajsetkrat večja od optične gostote samega ekstrakcijskega topila.
It should be documented that the tissue treated with NC is stable in culture (provide similar viability measurements) for the duration of the test exposure period.
Zabeležiti je treba, da je negativno kontrolno tkivo, kar zadeva celične kulture, med testiranjem stabilno (podobne meritve viabilnosti celic).
Barrier function
Pregradna funkcija
The stratum corneum and its lipid composition should be sufficient to resist the rapid penetration of cytotoxic marker substances, e.g. SDS or Triton X-100, as estimated by IC50 or ET50.
Rožena plast in njena maščobna sestava morata zadostovati za zaustavitev hitre penetracije citotoksičnih označevalcev, npr. SDS ali Triton X-100, kot se ocenjuje z IC50 ali ET50.
Morphology
Morfologija
Histological examination of the reconstructed skin/epidermis should be performed by appropriately qualified personnel demonstrating human skin/epidermis-like structure (including multilayered stratum corneum).
Histološki pregled rekonstruirane kože/pokožnice za prikaz strukture človeške kože/pokožnici podobne strukture (vključno z večplastno roženo plastjo) mora opraviti ustrezno usposobljeno osebje.
The results of the method using a specific model should demonstrate reproducibility over time, preferably by an appropriate batch control (benchmark) substance (see Appendix).
Rezultati metode s posebnim modelom morajo dokazati, da se koža sčasoma obnovi, najbolje z ustrezno (primerjalno) snovjo kontrolne serije (glej Dodatek).
Quality controls (QC) of the model
Nadzor kakovosti modela
Each batch of the epidermal model used should meet defined production release criteria, among which those for viability (paragraph 1.6.1.2.1) and for barrier function (paragraph 1.6.1.2.2) are the most relevant.
Vsaka uporabljena serija modela pokožnice mora izpolnjevati opredeljena merila za sprostitev proizvodnje, med katerimi so najpomembnejša merila v zvezi z viabilnostjo celic (odstavek 1.6.1.2.1) in pregradno funkcijo (odstavek 1.6.1.2.2).
An acceptability range (upper and lower limit) for the IC50 or the ET50 should be established by the skin model supplier (or investigator when using an in-house model).
Razpon sprejemljivosti (zgornja in spodnja meja) za IC50 ali ET50 mora določiti dobavitelj modela kože (ali raziskovalec ob uporabi lastnega modela).
The barrier properties of the tissues should be verified by the laboratory after receipt of the tissues.
Značilnosti pregrade tkiv se morajo preveriti v laboratoriju po prejetju tkiv.
Only results produced with qualified tissues can be accepted for reliable prediction of irritation effects.
Le rezultati, doseženi z ustreznimi tkivi, se štejejo kot zanesljivi za napovedi v zvezi z vplivi za draženje kože.
As an example, the acceptability ranges for the validated reference methods are given below.
Kot primer so spodaj navedeni razponi za validirane referenčne metode.
Examples of QC batch release criteria
Primeri meril za nadzor kakovosti serije
Lower acceptance limit
Spodnja meja sprejemljivosti
Mean of acceptance range
Vrednost razpona sprejemljivosti
Upper acceptance limit
Zgornja meja sprejemljivosti
Validated reference method 1 (18 hours treatment with SDS)
Validirana referenčna metoda 1 (18-urno testiranje z SDS)
Validated reference method 2 (1 % Triton X-100)
Validirana referenčna metoda 2 (1-odstotni Triton X100)
Application of the Test and Control Substances
Nanos testnih in kontrolnih snovi
A sufficient number of tissue replicates should be used for each treatment and for controls (at least three replicates per run).
Za vsako testiranje in kontrole je treba uporabiti zadostno število dvojnikov tkiv (vsaj tri dvojnike na vsako ponovitev testa).
For liquid as well as solid substances, sufficient amount of test substance should be applied to uniformly cover the skin surface while avoiding an infinite dose (see 1.2 Definitions), i.e. a minimum of 25 μL/cm2 or 25 mg/cm2 should be used.
Za tekoče in trdne snovi je treba uporabiti zadostno količino testne snovi, da se enakomerno prekrije površina kože, pri čemer mora biti odmerek natančno določen (glej 1.2 Opredelitve), tj. najmanj 25 μL/cm2 ali 25 mg/cm2.
For solid substances, the epidermis surface should be moistened with deionised or distilled water before application, to ensure good contact with the skin.
Da se zagotovi dober stik s kožo, je treba v primeru trdnih snovi površino pokožnice pred nanosom navlažiti z deionizirano ali destilirano vodo.
At the end of the exposure period, the test substance should be carefully washed from the skin surface with aqueous buffer, or 0,9 % NaCl.
Ob koncu časa izpostavljenosti je treba testno snov skrbno sprati s površine kože z vodnim puferjem ali 0,9-odstotnim NaCl.
Depending on the reconstructed human epidermis model used, the exposure period may vary between 15 to 60 minutes, and the incubation temperature between 20 and 37 °C.
Glede na uporabljeni model rekonstruirane človeške pokožnice lahko čas izpostavljenosti traja od 15 do 60 minut, inkubacijska temperatura pa meri od 20 do 37 °C.
For details, see the Standard Operating Procedures for the three methods (21, 22, 23).
Za podrobnejše informacije glej Standardne operativne postopke za te tri metode (21, 22, 23).
Concurrent NC and positive controls (PC) should be used for each study to demonstrate that viability (NC), barrier function and resulting tissue sensitivity (PC) of the tissues are within a defined historical acceptance range.
Negativna kontrola (NC) in pozitivna kontrola (PC) se morata sočasno uporabiti za vsako študijo, da se dokaže, da so viabilnost celic (negativna kontrola), pregradna funkcija in posledična občutljivost tkiva (pozitivna kontrola) v okviru v preteklosti opredeljenega razpona sprejemljivosti.
The suggested PC substance is 5 % aqueous SDS.
Predlagana snov za pozitivno kontrolo je 5-odstotni vodni SDS.
The suggested NC substances are water or phosphate buffered saline (PBS).
Predlagani snovi za negativno kontrolo sta voda ali fosfatni pufer s soljo (PBS).
Cell Viability Measurements
Meritve viabilnosti celic
The most important element of the test procedure is that viability measurements are not performed immediately after the exposure to the test substances, but after a sufficiently long post-treatment incubation period of the rinsed tissues in fresh medium.
Pri postopku testiranja je zelo pomembno, da se meritve viabilnosti celic ne opravljajo takoj po izpostavljenosti testnim snovem, ampak v svežih gojiščih po dovolj dolgi inkubacijski dobi po testiranju izpranih tkiv.
This period allows both for recovery from weakly irritant effects and for appearance of clear cytotoxic effects.
Ta čas omogoča izboljšanje stanja, kar zadeva šibke dražilne vplive, in pojav jasnih citotoksičnih vplivov.
During the test optimisation phase (9, 10, 11, 12, 13), a 42 hours post-treatment incubation period proved to be optimal and was therefore used in the validation of the reference test methods.
V fazi optimizacije testa (9, 10, 11, 12, 13) se je kot najugodnejša izkazala 42-urna inkubacijska doba po testiranju, zato se je uporabljala za validacijo referenčnih testnih metod.
The MTT conversion assay is a quantitative validated method which should be used to measure cell viability.
Analiza konverzije MTT je kvantitativna validirana metoda, ki jo je treba uporabljati za merjenje viabilnosti celic.
It is compatible with use in a three-dimensional tissue construct.
Ustreza uporabi tridimenzionalne konstrukcije tkiva.
The skin sample is placed in MTT solution of appropriate concentration (e.g. 0,3-1 mg/mL) for 3 hours.
Vzorec kože se za 3 ure potopi v ustrezno koncentracijo raztopine MTT (npr. 0,3–1 mg/mL).
The precipitated blue formazan product is then extracted from the tissue using a solvent (e.g. isopropanol, acidic isopropanol), and the concentration of formazan is measured by determining the OD at 570 nm using a bandpass of maximum ±30 nm.
S topilom (npr. izopropanolom, kislim izopropanolom) se izvleče oborina modrega formazana, potem pa se koncentracija formazana izmeri tako, da se določi OD pri valovni dolžini 570 nm ±30 nm.
Optical properties of the test substance or its chemical action on the MTT may interfere with the assay leading to a false estimate of viability (because the test substance may prevent or reverse the colour generation as well as cause it).
Optične značilnosti testne snovi ali njenega kemijskega delovanja na MTT lahko zato ovirajo analizo, zaradi česar je lahko ocena viabilnosti celic lažna (ker lahko testna snov prepreči obarvanje ali povzroči, da se razlikuje od običajnega).
This may occur when a specific test substance is not completely removed from the skin by rinsing or when it penetrates the epidermis.
To se lahko zgodi, če se določena testna snov s splakovanjem ne odstrani s kože v celoti ali če prodre v pokožnico.
If the test substance acts directly on the MTT, is naturally coloured, or becomes coloured during tissue treatment, additional controls should be used to detect and correct for test substance interference with the viability measurement technique.
Če testna snov deluje neposredno na MTT, je običajne barve ali se obarva med testiranjem tkiva, je treba izvesti dodatne kontrole, da se ugotovijo in odpravijo interference testne snovi z merilno tehniko viabilnosti.
For detailed description of how to test direct MTT reduction, please consult the test method protocol for the validated reference methods (21, 22, 23).
Za podrobnejši opis načina neposrednega testiranja zmanjšanja MTT glej protokol testne metode za validirane referenčne metode (21, 22, 23).
Non-specific colour (NSC) due to these interferences should not exceed 30 % of NC (for corrections).
Nespecifična barva zaradi teh interferenc ne sme preseči 30-odstotne negativne kontrole (za popravke).
If NSC > 30 %, the test substance is considered as incompatible with the test.
Če je NSC > 30 %, se šteje, da je testna snov neskladna s testom.
Assay Acceptability Criteria
Merila za sprejemljivost analize
For each assay using valid batches (see paragraph 1.6.1.2.5), tissues treated with the NC should exhibit OD reflecting the quality of the tissues that followed all shipment and receipt steps and all the irritation protocol process.
Za vsako analizo, pri kateri se uporabljajo ustrezne/veljavne serije (glej odstavek 1.6.1.2.5), morajo negativna kontrolna tkiva izražati optično gostoto, iz katere je razvidna kakovost tkiv, pri katerih so se upoštevale vse zahtevane stopnje in vsi postopki protokola v zvezi z razdraženostjo kože.
The OD values of controls should not be below historical established lower boundaries.
Vrednosti kontrol optične gostote ne smejo biti nižje kot v preteklosti postavljene nižje meje.
Similarly, tissues treated with the PC, i.e. 5 % aqueous SDS, should reflect the sensitivity retained by tissues and their ability to respond to an irritant substance in the conditions of each individual assay (e.g. viability ≤ 40 % for the validated reference method 1, and ≤ 20 % for the validated reference method 2).
Podobno morajo tkiva s PC, tj. 5-odstotnim vodnim SDS, kazati občutljivost tkiv in njihovo sposobnost odzivanja na dražilne snovi v pogojih vsake posamezne analize (na primer viabilnost celic ≤ 40 % za validirano referenčno metodo 1 in ≤ 20 % za validirano referenčno metodo 2).
Associated and appropriate measures of variability between tissue replicates should be defined (e.g. if standard deviations are used they should be ≤ 18 %).
S tem povezane in ustrezne meritve spremenljivosti med dvojniki tkiv je treba opredeliti (npr. v primeru uporabe standardnih odmikov mora biti njihova vrednost ≤ 18 %).
For each treatment, data from individual replicate test samples (e.g. OD values and calculated percentage cell viability data for each test substance, including classification) should be reported in tabular form, including data from repeat experiments as appropriate.
Za vsako testiranje je treba podatke o posameznih vzorcih dvojnikov testov (npr. vrednosti optične gostote in podatke o izračunanem deležu viabilnosti celic za vsako testno snov, vključno z razvrščanjem) sporočiti v tabeli, po potrebi vključno s podatki o ponovljenih poskusih.
In addition means ± standard deviation for each trial should be reported.
To pomeni, da je treba sporočiti standardni odmik ± za vsak poskus.
Observed interactions with MTT reagent and coloured test substances should be reported for each tested substance.
Za vsako testirano kemikalijo je treba sporočiti opaženo medsebojno vplivanje z reagentom MTT in obarvanimi testnimi snovmi.
The OD values obtained with each test sample can be used to calculate the percentage of viability compared to NC, which is set at 100 %.
Vrednosti OD, pridobljene z vsakim testnim vzorcem, se lahko uporabijo za izračun deleža viabilnosti celic NC, ki je določena na 100 %.
The cut-off value of percentage cell viability distinguishing irritant from non-classified test substances and the statistical procedure(s) used to evaluate the results and identify irritant substances, should be clearly defined and documented, and proven to be appropriate.
Mejna vrednost deleža viabilnosti celic, ki dražilne testne snovi loči od nerazvrščenih, ter statistični postopek(-ki), ki se uporablja(-jo) za ocenjevanje rezultatov ter določanje dražilnih snovi, morata(-jo) biti natančno opredeljena(-i) in zabeležena(-i) ter dokazana(-i) kot ustrezna(-i).
The cut-off values for the prediction of irritation associated with the validated reference methods is given below:
Mejne vrednosti za napovedovanje rezultatov v zvezi z razdraženostjo kože, ki zadevajo validirane referenčne metode, so navedene spodaj.
The test substance is considered to be irritant to skin in accordance with UN GHS category 2:
Testna snov se v skladu s kategorijo 2 GHS ZN šteje kot dražilna za kožo:
if the tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50 %.
če je viabilnost tkiva po izpostavljenosti in inkubaciji po testiranju manjša kot ali enaka (≤) 50 %.
The test substance is considered to have no category:
Testna snov se šteje kot snov brez kategorije:
if the tissue viability after exposure and post-treatment incubation is more than (>) 50 %.
če je viabilnost tkiva po izpostavljenosti in inkubaciji večja (>) kot 50 %.
The test report should include the following information:
Poročilo o testu mora vključevati naslednje informacije:
Test and Control Substances:
Testne in kontrolne snovi:
chemical name(s) such as IUPAC or CAS name and CAS number, if known,
kemijsko(-a) ime(-na), npr. IUPAC ali ime CAS in številko CAS, če je znana,
purity and composition of the substance (in percentage(s) by weight),
čistost in sestavo snovi (v odstotku(-ih) mase),
physical-chemical properties relevant to the conduct of the study (e.g. physical state, stability and volatility, pH, water solubility if known),
fizikalno-kemijske značilnosti, ki so pomembne za izvedbo študije (npr. agregatno stanje, obstojnost in hlapnost, pH, topnost v vodi, če so znane),
treatment of the test/control substances prior to testing, if applicable (e.g. warming, grinding),
obdelava testnih/kontrolnih snovi pred testiranjem, če je to smiselno (npr. segrevanje, mletje),
storage conditions.
pogoji skladiščenja.
Test Conditions:
Testni pogoji:
calibration information for measuring device, and bandpass used for measuring cell viability (e.g. spectrophotometer),
podatki o umerjanju za merilno napravo, s katero je bila izmerjena viabilnost celic (npr. spektrofotometer),
viability;
viabilnost;
barrier function;
pregradno funkcijo;
morphology;
morfologijo;
reproducibility and predictivity;
ponovljivost in napovedljivost;
quality controls (QC) of the model;
nadzor kakovosti modela:
details of the test procedure used,
podrobnejše informacije o uporabljenem postopku testiranja,
test doses used, duration of exposure and post treatment incubation period,
uporabljeni testni odmerki, čas izpostavljenosti in inkubacijska doba po testiranju,
description of any modifications of the test procedure,
opis vseh sprememb postopka testiranja,
acceptability of the QC data with reference to historical batch data;
sprejemljivost podatkov o nadzoru kakovosti s sklicevanjem na podatke o seriji, pridobljene v preteklosti;
acceptability of the positive and negative control values with reference to positive and negative control means and ranges,
sprejemljivost vrednosti pozitivne in negativne kontrole s sklicevanjem na pozitivna ter negativna kontrolna sredstva in razpone,
description of evaluation criteria used including the justification for the selection of the cut-off point(s) for the prediction model.
opis uporabljenih meril za ocenjevanje, vključno z zagovorom izbire mejne(-ih) vrednosti za napovedni model.
tabulation of data from individual test samples,
evidentiranje podatkov posameznih testnih vzorcev,
description of other effects observed.
opis vseh ostalih opaženih vplivov.
Available at: http://guidance.echa.europa.eu/docs/guidance_document/information_requirements_en.htm?time=1232447649.
Na voljo na spletni strani: http://guidance.echa.europa.eu/docs/guidance_document/information_requirements_en.htm?time=1232447649.
Available under Download Study Documents, at: http://ecvam.jrc.ec.europa.eu.
Na voljo pod Download Study Documents na spletni strani: http://ecvam.jrc.ec.europa.eu.
Toxicology in Vitro 15, 57-93.
Results and evaluation by the Management Team, Toxicology in Vitro, 15, 57–93.
ESAC statement on updated EpiDerm and similar SkinEthic assays. 5 November 2008.
ESAC statement on updated EpiDerm and similar SkinEthic assays, 5. november 2008.
Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC.
Uredba (ES) št. 1907/2006 Evropskega parlamenta in Sveta z dne 18. decembra 2006 o registraciji, evalvaciji, avtorizaciji in omejevanju kemikalij (REACH), o ustanovitvi Evropske agencije za kemikalije ter spremembi Direktive 1999/45/ES ter razveljavitvi Uredbe Sveta (EGS) št. 793/93 in Uredbe Komisije (ES) št. 1488/94 ter Direktive Sveta 76/769/EGS in direktiv Komisije 91/155/EGS, 93/67/EGS, 93/105/ES in 2000/21/ES.
Will be available under Download Study Documents, at: http://ecvam.jrc.ec.europa.eu.
Kmalu na voljo pod Download Study Documents na spletni strani: http://ecvam.jrc.ec.europa.eu.
FRESHWATER ALGAE AND CYANOBACTERIA, GROWTH INHIBITION TEST
SLADKOVODNE ALGE IN CIANOBAKTERIJE, PRESKUS ZAVIRANJA RASTI
This method is equivalent to OECD TG 201 (2006) (1).
Ta metoda je enakovredna OECD TG 201 (2006)(1).