commiss direct ec april adapt technic progress time council direct eec approxim law regul administr provis relat classif packag label danger substanc text eea relev commiss european commun regard treati establish european commun regard council direct eec june approxim law regul administr provis relat classif packag label danger substanc amend european parliament council direct ec articl thereof annex direct eec lai method determin physico chemic properti toxic ecotox substanc prepar adapt annex technic progress articl council direct eec novemb approxim law regul administr provis member state protect anim experiment scientif purpos experi entail anim perform scientif satisfactori method obtain result sought practic commiss intend introduc annex direct eec altern test method entail anim order make test chemic articl direct eec measur provid direct accord opinion committe adapt technic progress direct elimin technic barrier trade danger substanc prepar adopt direct articl text annex ii direct ad part annex direct eec articl member state bring forc law regul administr provis compli direct octob latest forthwith inform commiss thereof member state adopt provis refer direct accompani refer occas offici public member state determin refer made member state commun commiss main provis nation law adopt field cover direct correl tabl direct nation provis adopt articl direct enter forc dai public offici journal european commun articl direct address member state brussel april commiss margot wallström member commiss adopt adapt oj oj oj annex quot skin corros method introduct vitro test skin corros rat skin transcutan electr resist ter assai test emploi human skin model endors scientif valid european centr valid altern method ecvam joint research centr european commiss ecvam valid studi demonstr test reliabl discrimin skin corros corros test protocol base human skin model enabl correct distinct degre corros effect sever skin corros skin corros descript procedur test choic test depend specif requir prefer user gener introduct part definit skin corros product irrevers tissu damag skin applic test materi refer substanc point principl test method rat skin ter assai test materi appli hour epiderm surfac skin disc pelt human kill young rat corros materi identifi abil produc loss normal stratum corneum integr barrier function measur reduct inher ter threshold level kω irrit irrit materi reduc ter threshold level dye bind step incorpor test procedur surfact neutral organ definit refer reduc number fals posit result obtain specif chemic type descript test method rat skin ter assai anim young dai rat wistar compar strain requir prepar skin disc dorsal flank hair carefulli remov small anim clipper anim wash care wipe whilst submerg area antibiot solut streptomycin penicillin chloramphenicol amphotericin concentr effect inhibit bacteri growth anim wash antibiot fourth dai wash dai anim older dai pelt prepar prepar skin disc anim human kill dorsal skin anim remov strip excess fat carefulli peel skin pelt end polytetrafluoroethylen ptfe tube ensur epiderm surfac contact tube rubber ring press fit end tube hold skin place excess tissu trim tube ring dimens shown figur rubber ring carefulli seal end ptfe tube petroleum jelli tube support spring clip insid receptor chamber contian magnesium sulphat solut mm figur test procedur applic test materi liquid test substanc µl appli epiderm surfac insid tube figur test solid materi suffici amount solid appli disc ensur surfac epidermi cover deionis water µl ad top solid tube gentli agit test substanc maximum contact skin solid achiev warm melt test substanc grind produc granular materi powder skin disc test substanc test substanc appli hour test substanc remov wash jet tap water materi remov remov test substanc solidifi tube facilit jet wash warm water approxim ter measur ter measur low voltag altern current databridg aim equival prior measur electr resist surfac tension skin reduc ad suffici volum ethanol cover epidermi second ethanol remov invert tube tissu hydrat addit ml magnesium sulphat solut mm databridg electrod side skin disc resist measur kω skin disc figur electrod dimens length electrod expos crocodil clip shown figur thick electrod clip rest top ptfe tube resist measur ensur consist length electrod submerg magnesium sulphat solut outer thin electrod posit insid receptor chamber rest bottom chamber distanc bottom spring clip bottom ptfe tube maintain constant figur distanc affect resist obtain note measur resist greater kω due test substanc coat epiderm surfac sking disc remov coat attempt seal ptfe tube glove thumb shake approxim second magnesium sulphat solut discard resist measur repeat fresh magnesium sulphat ter result accept condit concurr posit neg control valu fall accept rang method suggest control substanc accept resist rang methodolog apparatu gt tabl gt modifi procedur surfact neutral organ ter valu test substanc surfact neutral organ equal kω assess dye penetr carri tissu procedur determin result fals posit sulforhodamin dye applic remov initi treatment test substanc µl dilut sulforhodamin dye distil water appli epiderm surfac skin disc hour skin disc jet wash tap water room temperatur approxim second remov excess unbound dye skin disc carefulli remov ptfe tube vial ml glass scintil vial deionis water ml vial agit gentli minut remov excess unbound dye rins procedur repeat skin disc remov vial ml sodium dodecyl sulphat sd distil water incub overnight incub skin disc remov discard remain solut centrifug minut rel centrifug forc sim ml sampl supernat dilut ml ml sd distil water optic densiti od solut measur approxim nm calcul dye content sulforhodamin dye content disc calcul od valu sulforhodamin dye molar extinct coeffici nm molecular weight sulforhodamin dye content determin skin disc dye content calcul replic dye bind result accept condit concurr control valu fall accept rang method suggest accept dye content rang control substanc methodolog apparatu gt tabl gt addit inform test substanc appli skin disc shorter period hour identifi materi sever corros valid studi ter assai found overestim corros potenti test chemic applic skin disc hour enabl correct identif corros corros hour applic properti dimens test apparatu experiment procedur influenc ter valu obtain kω corros threshold develop data obtain specif apparatu procedur method threshold control valu appli test condit alter significantli recommend methodolog resist threshold calibr test seri refer standard chosen chemic valid studi principl test method human skin model assai test materi appli topic hour dimension human skin model compris reconstruct epidermi function stratum corneum corros materi identifi abil produc decreas cell viabil determin mtt reduct assai defin threshold level exposur period principl essai accord hypothesi chemic corros penetr stratum corneum diffus eros suffici cytotox cell death underli cell layer descript test method human skin model assai human skin model human skin model sourc meet criteria model function stratum corneum underli layer live cell barrier function stratum corneum adequ shown demonstr model resist cytotox applic substanc cytotox cell pass stratum corneum model shown give reproduc result defin experiment condit viabil live cell model suffici high discrimin posit neg control substanc cell viabil measur amount mtt reduct od exposur neg control substanc fall accept limit model similarli cell viabil valu posit control substanc rel neg control fall limit importantli predict model shown meet intern valid standard test procedur applic test materi liquid materi suffici test substanc appli cover skin surfac minimum µl cm solid materi suffici test substanc appli cover skin moisten ensur good contact skin solid ground powder applic applic method shown adequ wide rang chemic type end exposur period test materi carefulli wash skin surfac salin solut cell viabil measur quantit valid method measur cell viabil frequent assai mtt reduct shown give accur reproduc result laboratori skin disc mtt solut mg ml hour precipit blue formazan product extract solvent extract concentr formazan measur determin od wavelenght nm addit inform skin model exact protocol exposur time wash procedur major impact cell viabil result recommend methodolog predict model calibr test seri refer standard chosen chemic ecvam valid studi critic method shown reproduc laboratori wide rang chemic accord intern standard minimum method meet criteria scientif valid defin previous result valid studi publish peer review scientif journal data treatment result rat skin ter assai resist valu kω test materi posit neg control standard refer chemic report tabular form includ data replic repeat experi valu classif deriv human skin model essai od valu calcul percentag cell viabil data test materi posit neg control standard refer chemic report tabular form includ data replic repeat experi valu classif deriv evalu interpret result rat skin ter assai ter obtain test substanc greater kω corros ter equal kω test substanc surfact neutral organ corros surfact neutral organ give ter valu equal kω dye penetr carri disc dye content greater equal disc dye content hcl posit control obtain concurr test substanc true posit corros disc dye content disc content hcl posit control obtain concurr test substanc fals posit corros human skin model assai neg control od repres cell viabil od valu obtain test sampl calcul percentag viabil rel neg control cut percentag cell viabil distinguish corros corros test materi discrimin corros class defin predict model method valid subsequ valid studi show cut report test report test report includ inform test substanc identif data physic natur relev physicochem properti similar inform provid refer substanc test condit detail test procedur descript justif modif result tabul resist valu ter assai percentag cell viabil valu human skin model assai test materi posit neg control standard refer chemic includ data replic repeat experi valu descript effect observ discuss result conclus refer ecvam ecvam new amp view atla pp fentem archer ball botham curren earl esdail holzhutt amp liebsch ecvam intern valid studi vitro test skin corros result evalu manag team toxicolog vitro pp barratt brantom fentem gerner walker amp worth ecvam intern valid studi vitro test skin corros select distribut test chemic toxicolog vitro pp oliv pemberton amp rhode vitro skin corros test modif valid food amp chemic toxicolog pp botham hall dennett mccall baskett whittl cheeseman esdail amp gardner skin corros test vitro result interlaboratori trial toxicolog vitro pp worth fentem ball botham curren earl esdail amp liebsch evalu propos oecd test strategi skin corros atla pp botham chamberlain barratt curren esdail gardner gordon hildebrand lewi liebsch logemann osborn ponec regnier steil walker amp ball prevalid studi vitro skin corros test report recommend ecvam workshop atla pp figur gt pic file quot en tif quot gt figur gt pic file quot en tif quot gt quot annex ii quot phototox vitro nru phototox test method introduct phototox defin toxic respons elicit exposur skin chemic subsequ exposur light induc similarli skin irradi system administr chemic inform deriv vitro nru phototox test serv identifi phototox potenti test substanc exist absenc hazard aris test substanc associ exposur uv visibl light toxicolog endpoint vitro test determin photocytotox induc combin action chemic light compound phototox vivo system applic distribut skin compound act photoirrit topic applic skin identifi test vitro nru phototox test develop valid joint eu colipa project establish valid vitro altern vivo test oecd workshop recommend vitro tier test approach phototox assess result vitro nru phototox test compar acut phototox photoirrit effect vivo anim human test shown give excel predict effect test design predict advers effect aris combin action chemic light photogenotox photoallergi photocarcinogen chemic show specif properti react posit vitro nru phototox test addit test design permit assess phototox potenc sequenti approach phototox test chemic set appendix definit irradi intens ultraviolet uv visibl light incid surfac measur mw cm dose light quantiti intens time ultraviolet uv visibl radiat incid surfac express joul surfac area cm uv light waveband design recommend cie commiss international de eclairag uva nm uvb nm uvc nm design divis uvb uva nm uva divid uv uv divis made nm cell viabil paramet measur total activ cell popul uptak vital dye neutral red cellular lysosom depend endpoint measur test design correl total number vital cell rel cell viabil cell viabil express relat neg solvent control test procedur uv uv treat test chemic predict model algorithm transform result toxic test predict toxic potenti present test guidelin pif mpe transform result vitro nru phototox test predict phototox potenti pif photo irrit factor factor gener compar equal effect cytotox concentr ec test chemic obtain absenc uv presenc uv cytotox irradi uva vi light mpe photo effect measur deriv mathemat analysi complet shape concentr respons curv obtain absenc uv presenc uv noncytotox irradi uva vi light phototox acut toxic respons elicit exposur skin chemic subsequ exposur light induc similarli skin irradi system administr chemic photoirrit speci term phototox describ phototox reaction produc skin exposur chemic topic oral photox reaction lead specif cell damag sunburn reaction photoallergi acquir immunolog reactiv occur treatment chemic light induct period week skin reactiv demonstr photogenotox genotox respons observ genet endpoint elicit exposur cell genotox dose uv visibl light genotox chemic photocarcinogen carcinogen induc repeat applic light chemic term photo carcinogenesi uv induc tumorigenesi enhanc chemic refer substanc posit control chemic chlorpromazin concurr test assai newli establish nru phototox test recommend refer chemic subset chemic interlaboratori trial present test initi consider type chemic report induc phototox effect common featur abil absorb light energi sunlight region law photochemistri grotthau draper law photoreact requir suffici absorpt light quanta biolog test present test guidelin consid uv vi absorpt spectrum test chemic determin oecd test guidelin molar extinct absorpt coeffici litr mol cm chemic photoreact potenti test vitro nru phototox test biolog test advers photochem effect appendix principl test method mechan identifi absorpt light chemic chromophor result phototox respons result cell damag vitro nru phototox test base comparison cytotox chemic test presenc absenc exposur cytotox dose uva vi light cytotox test express concentr depend reduct uptak vital dye neutral red nr hour treatment test chemic irradi balb cell maintain cultur format monolay plate test chemic preincub concentr chemic plate expos cytotox uva vi light dose cm uva uv experi plate dark uv experi plate treatment medium replac cultur medium incub cell viabil determin neutral red uptak nru rel cell viabil express percentag untreat neg control calcul test concentr predict phototox potenti concentr respons obtain presenc uv absenc uv irradi compar ec level concentr inhibit cell viabil cf untreat control qualiti criteria uva sensit cell histor data cell regularli check sensit uva cell seed densiti vitro nru phototox test irradi dai uva dose cm cell viabil determin dai nru assai cell meet qualiti criteria viabil irradi cm uva viabil dark control highest uva dose cm viabil dark control check repeat passag cell uva sensit neg control cell current test test meet qualiti criteria neg control cell earl balanc salt solut ebss dimethylsulfoxid dmso ethanol etoh uva experi show viabil irradi cell solvent concurr dark experi uva viabil neg control absolut optic densiti od nru measur nr extract neg control cell seed grown normal doubl time dai assai test meet accept criteria od nru untreat control amp gt posit control phototox chemic test concurr vitro nru phototox test chlorpromazin cpz posit control eu colipa valid studi recommend cpz test standard protocol vitro nru phototox test test accept criteria defin cpz irradi uva ec µg ml cpz irradi uva ec µg ml photo irrit factor pif shift ec phototox chemic suitabl chemic class solubl characterist test chemic evalu concurr posit control place cpz case base histor data rang ec valu pif mpe photo effect adequ defin accept criteria test descript test method prepar cell perman mous fibroblast cell line balb clone atcc ecacc valid studi recommend cell cell line successfulli test protocol cultur condit adapt specif cell equival demonstr cell check regularli absenc mycoplasma contamin result check satisfactori uva sensit cell increas number passag balb cell lowest obtain passag number prefer import uva sensit balb cell regularli check qualiti control procedur guidelin media cultur condit cultur media incub condit routin cell passag test procedur balb cell dmem supplement born calf serum mm glutamin penicillin streptomycin humidifi incub import cell cultur condit ensur cell cycl time normal histor rang cell cell line prepar cultur cell frozen stock cultur seed cultur medium densiti subcultur vitro nru phototox test phototox test cell seed cultur medium densiti cultur reach confluenc end test cell viabil determin seed cell balb cell grown plate cell recommend cell densiti test chemic cell seed ident separ plate concurr test procedur ident cultur condit time period plate irradi uva vi dark uva vi metabol activ metabolis system gener requir vitro test predict genotox carcinogen potenti case phototoxicolog chemic metabol transform need chemic act phototoxin vivo vitro consid scientif justifi present test perform metabol activ system test chemic prepar test chemic freshli prepar immedi prior stabil data demonstr accept storag prepar red light requir rapid photodegrad occur test chemic dissolv buffer salt solut earl balanc salt solut ebss phosphat buffer salin pb avoid interfer irradi free protein compon light absorb ph indic colour test chemic limit solubl water dissolv solvent fold desir final concentr dilut buffer salt solut solvent present constant volum cultur neg control concentr test chemic dimethylsulphoxid dmso ethanol etoh recommend solvent solvent low cytotox aceton carefulli assess specif properti reaction test chemic quench phototox effect radic catch properti vortex mix sonic warm aid solubilis uv irradi prepar light sourc choic light sourc filter crucial factor phototox test uva visibl region photosensitis uvb relev directli highli cytotox increas cytotox fold nm criteria choic light sourc includ essenti requir light sourc emit wavelength absorb test chemic dose light achiev reason time suffici detect photosensitis wavelength dose emploi unduli deleteri test system includ emiss heat infra red region simul sunlight solar simul consid optim light sourc xenon arc dope mercuri metal halid arc solar simul advantag emit heat cheaper match sunlight perfect solar simul emit signific quantiti uvb suitabl filter attenu highli cytotox uvb wavelength vitro nru phototox test irradi spectrum practic devoid uvb uva uvb sim spectral irradi distribut filter solar simul valid studi vitro nru phototox test publish dosimetri intens light irradi regularli check phototox test suitabl broadband uv meter uv meter calibr sourc perform uv meter check purpos refer uv meter type ident calibr recommend ideal greater interv spectroradiomet measur spectral irradi filter light sourc check calibr broadband uv meter instrument requir skill oper appropri train person dose cm uva determin valid studi cytotox balb cell suffici potent excit weak phototox chemic achiev cm time period min irradi adjust mw cm cell line light sourc uva dose slightli adapt criteria deleteri cell suffici detect standard phototoxin time light exposur calcul gt pic file quot en ep quot gt test condit maximum concentr test chemic exce µg ml phototox chemic detect lower concentr higher concentr incid fals posit overpredict increas ph highest concentr test chemic satisfactori ph rang rang concentr chemic test presenc uva absenc uva light adequ determin preced rang finder experi rang intercept concentr seri adjust concentr respons curv suffici support experiment data geometr concentr seri constant dilut factor test procedur dai prepar cell suspens cell ml cultur medium dispens µl cultur medium peripher well tissu cultur microtit plate blank remain well dispens µl cell suspens cell ml cell test chemic prepar plate determin cytotox uva determin photocytotox uva incub cell form half confluent monolay incub period cell recoveri adher exponenti growth dai incub decant cultur medium cell wash µl ebss pb add µl ebss pb concentr test chemic solvent neg control appli concentr test chemic incub cell test chemic dark minut perform uva part assai irradi cell room temperatur minut lid plate mw cm uva cm ventil fan prevent condens lid duplic plate uva room temperatur dark box min uva exposur time decant test solut wash µl ebss pb replac ebss pb cultur medium incub overnight dai microscop evalu examin cell phase contrast microscop record morpholog cell due cytotox effect test chemic check recommend exclud experiment error record evalu cytotox phototox neutral red uptak test wash cell µl prewarm ebss pb remov wash solut gentl tap add µl nr medium incub humidifi atmospher incub remov nr medium wash cell µl ebss pb decant blot ebss pb total option centrifug revers plate add µl nr desorb solut freshli prepar ethanol acet acid shake microtit plate rapidli microtit plate shaker min nr extract cell form homogen solut measur optic densiti nr extract nm spectrophotomet blank refer save data file format ascii subsequ analysi data qualiti quantiti data data permit meaning analysi concentr respons obtain presenc absenc uva vi irradi cytotox found concentr rang intercept individu concentr set fit curv experiment data due fact test chemic cytotox defin limit concentr µg ml dark experi uva highli cytotox irradi uva concentr rang test part experi differ order magnitud fulfil requir adequ data qualiti cytotox found part experi uva uva test great intercept singl dose highest concentr suffici requir verif clear posit result perform repeat experi addit clear neg result verifi provid test chemic test suffici high concentr case main experi support rang find preliminari experi suffici test borderlin result cut line predict model repeat verif repeat test consid variat experiment condit import achiev clear result kei variabl test prepar solut test chemic variat condit solvent tritur sonic relev repetit test altern variat pre irradi incub time consid shorter time relev water unstabl chemic treatment result concentr test chemic reflect inhibit cellular nru ec determin appli linear regress procedur prefer hill function logist regress concentr respons data fit procedur ec calcul qualiti fit appropri check altern graphic fit method calcul ec case probabl paper recommend scale log scale probit case concentr respons function linear transform evalu result predict model predict model version photo irrit factor pif presenc uva absenc uva light complet concentr respons curv obtain photo irrit factor pif calcul mean formula gt pic file quot en ep quot gt pif amp lt predict phototox potenti pif amp gt predict phototox potenti chemic cytotox uva cytotox test uva pif calcul result phototox potenti case amp gt pif calcul uv cytotox test perform highest test concentr cmax calcul amp gt pif gt pic file quot en ep quot gt amp gt pif obtain amp gt predict phototox potenti ec uv ec uv calcul due fact chemic show cytotox highest test concentr phototox potenti case formal pif characteris result gt pic file quot en ep quot gt pif obtain predict phototox potenti case concentr achiev vitro nru phototox test carefulli consider predict phototox potenti predict model version photo effect mpe altern version model predict phototox potenti appli develop data eu colipa valid studi test blind condit subsequ studi vitro phototox uv filter chemic model overcom limit pif model case ec obtain model photo effect mpe measur base comparison complet concentr respons curv applic mpe model special comput softwar develop humboldt univers berlin obtain free charg interpret result posit result vitro nru phototox test pif amp gt mpe amp gt test substanc phototox potenti result obtain concentr µg ml test chemic act phototoxin exposur condit vivo posit result obtain highest test concentr µg ml consider assess hazard phototox potenc includ data penetr absorpt accumul chemic skin test chemic confirmatori altern test human vitro skin model neg result vitro nru phototox test pif amp lt mpe amp lt test substanc phototox cultur mammalian cell condit case chemic test highest concentr µg ml neg result chemic phototox potenti phototox vivo consid case ident concentr toxic respons ec uv ec uv obtain lower concentr interpret data contrast toxic demonstr uv uv aqueou solubl limit concentr valu µg ml compat test substanc assai question confirmatori test consid vitro skin model vivo skin model vivo test report test report test report includ inform test chemic identif data ca physic natur puriti physicochem properti relev conduct studi stabil photost solvent justif choic solvent solubl test chemic solvent percentag solvent present treatment medium ebss pb cell type sourc cell absenc mycoplasma number cell passag uva sensit cell determin irradi equip vitro nru phototox test test condit incub treatment type composit cultur medium incub condit concentr temperatur humid durat incub pre treatment post treatment test condit treatment chemic rational select concentr test chemic presenc absenc uv vi irradi case limit solubl test chemic absenc cytotox rational highest concentr test type composit treatment medium buffer salt solut durat chemic treatment test condit irradi rational select light sourc spectral irradi characterist light sourc transmiss absorpt characterist filter characterist radiomet detail calibr distanc light sourc test system uva irradi distanc express mw cm durat uv vi light exposur uva dose irradi time express cm temperatur emploi cell cultur irradi cell cultur concurr dark test condit nru test composit nr medium durat nr incub incub condit concentr temperatur humid nr extract condit extract durat wavelength spectrophotometr read nr optic densiti wavelength refer content spectrophotomet blank result cell viabil obtain concentr test chemic express percent viabil control concentr respons curv test chemic concentr rel cell viabil obtain concurr uva uva experi data analysi concentr respons curv comput calcul ec uva ec uva comparison concentr respons curv obtain presenc absenc uva vi irradi calcul photo irrit factor pif calcul photo effect mpe classif phototox potenti test accept criteria concurr neg control absolut viabil optic densiti nr extract irradi irradi cell histor data neg control standard deviat test accept criteria concurr posit control ec uva ec uva pif posit control chemic histor data posit control chemic ec uva ec uva pif standard deviat discuss result conclus refer spielmann ball döring holzhütter kalweit klecak eplatteni liebsch lovel maurer moldenhau moor pape pfannbeck potthast de silva steil willshaw eec colipa project vitro phototox test result obtain balb cell phototox assai toxicolog vitro pp anon statement scientif valid nru pt test vitro test phototox european commiss joint research centr ecvam dgxi novemb atla pp spielmann ball dupui pape pechovitch de silva holzhütter clothier desol gerberick liebsch lovel maurer pfannenbeck potthast csato sladowski steil brantom eu colipa vitro phototox valid studi result phase ii blind trial part nru phototox test toxicolog vitro pp oecd test guidelin programm env mc chem tg final report oecd workshop harmonis valid accept criteria altern toxicolog test method oecd public offic pari lovel scheme vitro screen substanc photoallergen potenti toxicolog vitro pp santamaria prino list photodynam substanc research progress organ biolog medicin chemistri vol part north holland publish amsterdam pp xi xxxv spielmann lovel hölzle johnson maurer miranda pape sapora sladowski vitro phototox test report recommend ecvam workshop atla pp spike photosensit scienc photobiolog edit kc smith plenum press york edit pp borenfreund puerner toxic determin vitro morpholog alter neutral red absorpt toxicolog letter pp lambert warner kornhaus anim model phototox test dermatotoxicolog edit fn marzulli maibach publish taylor amp franci washington dc edit pp tyrrel pidoux action spectra human skin cell estim rel cytotox middl ultraviolet ultraviolet violet region sunlight epiderm keratinocyt cancer research pp zebet ecvam colipa standard oper procedur balb nru phototox test draft decemb liebsch approv march manag team eu colipa project vitro photoirrit spielmann ball dupui pape de silva holzhütter gerberick liebsch lovel pfannenbeck studi phototox potenti uv filter chemic annex vii eu direct eec nru vitro phototox test atla pp holzhütter quedenau mathemat model cellular respons extern signal journal biolog system pp holzhütter gener measur vitro phototox deriv pair dose respons curv predict vivo phototox chemic atla pp appendix gt pic file quot en tif quot gt quot addit detail found refer