April 2006- September 2006. Britta Lende Hansen. Specialist. Nova 8. Leveret af: Triolab AS. YaUensbækvej 35. 2605 Brøndby. 15. Oktober.

Gentofte Hospital Klinisk-biokemisk afdeling Validitetserklæring for NPU01446 P- Calcinm- ion(frit):stofk. og NPU04144 P- Calcium- ion(frit);stofk.(ph=7,4) målt på NOVA 8 Rapport V-046/02 2 eksemplarer Valideringsperiode: Arbejdsgruppe: Udstyr: Ibrugtagningsdato April 2006- September 2006 Britta Lende Hansen. Specialist Nova 8. Leveret af: Triolab AS. YaUensbækvej 35. 2605 Brøndby. 15. Oktober. 2006 Godkendelse af validitet: (lln<!crskrilt slempel og dalo) Ledn. Overlæge dr. med. Steen Stander, Klinisk-biokemisk afd. opg. 38 k. Gentofte Hospital ~:.,,,.. 2900 Hellerup C'~'-.<.. Tlf.: 39 77 31 20 Resume Vi har i afdelingen vurderet vigtigheden af at validere den intel111ediære impræcision, samt udført præsisionsprofil, som gør det muligt at estimere variationskoefficienter (CV) for prøver med varierende koncentration. Vi har også undersøgt den relative standard måleusikkerhed og lineariteten, samt lavet apparatursammenligning med ABL 725. Resultaterne af ovenstående dokumenterer, at laboratoriet anvender en analysemetode med en kvalitet, som opfylder kravene til analysen som er fastlagt, så de ud fra en lægelig vurdering vil opfylde rekvirentens behov. Med hensyn til metrologisk sporbarhed, analytisk specificitet, robusthed, afsmitning og måleinterval har vi valgt at anvende producentens oplysninger, da disse er CE- mærkede. V O~6IoI1Cll Side l af4

Gentofte Hospital Klinisk-biokemisk afdeling Validitetserklæring for NPUOt446 P- Calcium- ion(frit);stotl<. og NPU04144 P- Calcium- ion(frit);stofk.(ph~7,4) målt på NOVA 8 Rapport V-046/02 2 eksemplarer Apparatursammenligning Korrekthed Konklusion Ved metodesammenligning foretaget overfor ABL 725, P- Calcium- ion(frit); stofk. findes resultater. der er meget ens. Apparaturet måler forskellig ph på ABL725 og Nova 8, hvilket giver en forskel på P- Calciurnion(frit),stofk(pH=7A). Efter kørsel afprøver i forskelligt niveau udregnes differensen mellem de 2 målinger og en gennemsnitlig afvigelse udregnes og lægges ind i Nova 8 som en værdi, der trækkes fra alle ph målinger på Nova 8. Herefter passer de to apparater med hinanden på ph og derfor vil den ph- korrigerede Ioniseret Calcium også passe med ABL725. Korrekthed er kort i 3 niveauer Niveau 1: 1,71 ± 0,08 rnmolll Niveau 2: 1,37±0,07rnmol/L Niveau 3: 1,09 ± 0.05 mmolll Alle målinger er indenfor de opgivne grænser. jf. Bilag 15-18. Krav Resultat Måleinterval p- Calcium- ion(frit)j stofk. 0.5-1.80 mmol/l 0,1-5,5 mmol/l Impræeisioncll er beregnet i 2 niveauer: p-caic ium-ion(fr it)stofk: 1,34 % (0,82 mmoill). 1.03 % (1,45 mmol/l). 1m præcision 1,5% P-Calciumion(frit);stofk.(pH=7,4): 0,93 % (0.97 mmol/l). 1,[5%( 1,66mmoI/L). Begge niveauer opfylder de stillede krav. Bilag 4-7 V-Q46 JonCa Side 2 af4

Gentofte Hospital Klinisk-biokemisk afdeling Validitetserklæring for NPUOl446 P- Calcium- ion(frit);stofk. og NPU04144 P- Calcium- ion(frit);stofk.(ph=7,4) målt på NOVA 8 Rapport V-U46/02 Ekspanderet relativ kombineret standard måleusikker- 5% hed 2 eksemplarer Ekspanderet relativ kombineret standard måleusikkerhed beregnes til: P-Calcium-ion(frit),stofk: 2,4 % (0,82 mmol/l) 2,0 % (1,45 mmol/l) P-Calciumion(fri t);slofk(ph~7,4): 1,9 % (0,97 mmol/l) 2,2 % Cl,66 mmol/l) Bilag 20-23 V-().l61on("a Side 3 af4

Gentofte Hospital Klinisk-biokemisk afdeling Validitetserklæring for NPU01446 P- Calcium- ion(frit);stofk. og NPU04144 P- Calcium- ion(frit);stofk.(ph=7,4) målt på NOVA 8 Rapport V-046!02 2 eksemplarer Registreringsoversigt Egenskab ~~ -". Resultaterne fremkommet ved ' o '" -'" - ~ ~.~.~ = - = ~ -~ Bilags- =. Dokumentation opbevares " ' ~ ~' i:~ nummer - Il = =;; ~= ~< li.~ - - ;r < = =~. =,. ~ < _. = _. Ol. f = 7 ", -~ Metodesammenligning x 1-3 Metrologisk sporbarhed x 8 Måleinterval x 12 Korrekthed x 15-18 Detektionsgrænse x Analytisk specificitet x 13 Linearitet x Il Intermediær impræcision x 4-7 Præcisionsprofil x 9-10 Robusthed Er ikke oplyst. Afsmitning x 14 Præanalytiske forhold x Metodeblad KBA er ikke udfærdiget end- 19 nu. Ekspanderet relativ kombine- ret standard måleusikkerhed x 20-23 Ekstern kvalitetssikring x Rum 117 (Weqas blodgas) Andre forhold Referenceinterval x 24... Dokumenthistorik --------1 Overført til ny skabelon V-046IonCa Side 4 af 4

~ E ~ 1,4 Gentofte Hospital Klinisk-biokemisk afdeling Bilag 1 P-Calcium-ion(frit};stofk Metodesammenligning ABL725 VS. Nova 8 1.8,-----------------------------="1 ~ y = 1,0213x - 0,0238 1,6 ~ zz 1.2 I n 32 I 1.0 """'------~------_-------_--------J '" ABL 72S 1m01ol/l. x-y - - - Lineær (y) I Differensplot (y.x som funktion af x) 0.2 0,15 0.1.................,...,............................. 0.05 O S 0.05 0.1 0.15 0.2...I n 32 +...~ +... + "'!. I l... -'" '-'" - -. -......... -..... -.... 50/... 113: IOo/...lIæ -50/...lIæ 100/...113: I Konklusion: De to apparater måler ens. Nova 8 kan umiddelbart erstatte AEL 725. Bilag IIGrafer 02 03 2007 Side l af l

Gentofte Hospital Klinisk biokemisk afdeling Bilag 2 P-CaLcium-ion(frit};stofk. (ph~7,4) Metodesammenligning ABL725 vs. Nova 8 Før ph-korrektion 1.8, 1.7, Y~ I,1392x - 0,0556,,, 1,6,, 1,5,..., ~ 1,4,, I--/~ 1,3 -.. ~ ',,r I > o 1,2, z,,. -. -. -. -. _I 1.1,,,,,,,," 0,9,, 0,8 ;;} ~ - " N, ~. ~.., ABL 72S I mmol/l I n 25 1--x;;y -. -. Referenceinterval - - - Lineær (y) I y '", ~. " Differensplot (y-x som funktion af x)... '. 0,2,------------------------------,... 1 I,...1...~ " ' '1 0,1......... i*... j,, I I o o -0,1 1 n-25 I f i I j" o" o ~o I I o, 0 0 00. o..0.2 "- I o o..j 5o/o-væ... IO"/o-væ.5o/o-væ ' o. o. -IO"/o-væ. Referenceinterval! Konklusion; Før ph korrektion ses der for stor forskel på de apparaturer. Bilag 2JGrafcr 02 03 2007 Side [ af l

Gentofte Hospital Klinisk-biokemisk afdeling Bilag 3 P-Calcium-ion(frit);stojk. (ph 7,4) Metodesammenligning ABL725 vs. Nova 8 Efter ph-korrektion 1,7 1,6 y ~ 1,0213. - 0,0249,, -~ 1,5 5 o 1.4 1,3 > z 1.2 1,1 I.._. _. _. _. _.I n 26 I '. ABL 725 f mmol/l 1--x=y - -. Referenceinterval - - - Lineær (y)i Differensplot (y-x som funktion af x) 0'2,----------------------------,.--.. -... I... -... -... -...-. ~... -.... -.. -... -..'.-..-... -..,-... -.. 0.1 - ' -.' -.' o +-----_~>+_--'.'+<...~.'.!...--,...,...,.,.'------~-----'.~--- < ~.-t ~ _ ~l --.. _~-.-.. --.. _ In- 26 1!, I -. '.- -.. -'" -'. '. --..-.... --.. -... -. 5%-væ. loo/o-væ 5o/o-væ. -10%-væ -. - - Refercnceinlerval Konklusion: Nova 8 er ph korrigeret med en værdi på -0,16. De to metoder måler ens - det vil sige Nova kan erstatte ABL 725. Bilag 3/Grafer 19 09 2007 Side l af l

Klinisk-biokemisk afdeling Gentofte Hospital 02-03-2007 Total impræcision bestemt v.hj.a. Patient prøve. Analyse: P-Calclum-Ion{frlt); sjojk. Reagens lot nr.: 1SJ96 Kalibrator 101 nr.: 15196 Målt på: Nova 8 1mmo/A. N., Dato Værdi I 17-05-2006 0,82 2 17-05-2006 0,S2 3 17-05-2006 0,82 4 17-05-2006 0,81 5 17-05-2006 0,81 6 17-05-2006 0,82 7 18-05-2006 0,8\ 8 18-05-2006 0,8\ 18-05-2006 0,8\ 10 18-05-2006 0,82 Il 18-05-2006 0,82 12 18-05-2006 0,82 13 19-05-2006 0,83 14 19-05-2006 0,84 15 19-05-2006 0,84 16 19-05-2006 0,83 '7 19-05-2006 0,83 18 19-05-2006 0,84 " 22-05-2006 O,Sl 20 22-05-2006 0,82 21 22-05-2006 0,84 22 22-05-2066 0,S4 Middel 0,82 sd 0,011 CV/% 1,34 Bilag 4

Klinisk biokemisk afdeling Gentofte Hospital 02 03 2007 Total impræcision bestemt v.bj.a. Patient prøve. Analyse: Reagens lot nr.: Kalibrator lot nr.: MAlt på: p- Clllc/um-lon(fr/t); $loj1c. 15196 15196 NON 6 JmmoJA. N., Dato Værdi I 17-05-2006 1,47 2 17-05-2006 1,48 3 17-05-2006 1,47 4 17-05-2006 1,46 5 17-05-2006 1,46 6 17-05-2006 1,48 7 18-05-2006 1,46 8 18-05-2006 1,46 9 18-05-2006 1,46 \O 18-05-2006 1,46 \I 18-05-2006 1,47 12 18-05-2006 1,46 13 19-056-06 1,45 14 19-05-2006 1,44 15 19-05-2006 1,44 16 19-05-2006 1,44 17 19-05-2006 1,45 18 19-05-2006 1,44 19 22-05-2006 1,43 20 22-05-2006 1,43 21 22-05-2006 1,43 22 22-05-2006 1,43 23 23-05-2006 1,44 24 23-05-2006 1,44 25 23-05-2006 1,44 26 23-05-2006 1,46 27 23-05 2006 1,46 28 23-05-2006 1,46 29 23-05-2006 1,44 Middel 1,45 sd 0,015 CV/% 1,03 Bilag 5

Klinisk-biokemisk afdeling 08 0] 2007 Amtssygehuset i Gentofte Total impræcision bestemt v.bj.a. Patient prøve Analyse: ReageM lot nr.: Kalibrator 101 nr.: Milt pi: P-CtIldllm-lolf(frlt); Slojl.(pH-1.4) 1JJ96 JjI96.\'(WtI lo I trurtdia N,. Dato Værdi I 17 05-2006 0,97 2 17 05-2006 0,97 3 17 05-2006 0,97 4 17 05-2006 0.97 5 11-05-2006 0,97 6 17-05-2006 0,98 7 18--05-2006 0,96 8 18--05-2006 0,96 9 18-05-2006 0,96 IO 18-05-2006 0,98 II 18-05-2006 0,98 12 18--05-2006 0,98 13 19 05-2006 0,97 14 19 05-2006 0,98 15 19 05-2006 0,98 16 19 05-2006 0,97 17 19-05-2006 0,97 18 19 05-2006 1,00 19 22 05-2006 0,97 20 22 05-2006 0,98 21 22 05-2006 0,98 22 22-05-2006 0,98 Middel 0,97 sd 0,0091 CV/% 0,932 Bilag 6

Klinisk-biokemisk afdeling 08-03-2007 Gentofte Hospital Analyse: Total impræcision bestemt v.bj.a. Patient prøve. P-Calcium- lon(frit); stofk.(ph:s.7.4) Reagens lot nr.: Kalibrator lot nr.: Målt på: 15196 15196 Nova l. i mmola N" Dato Værdi I 17-05-2006 1,64 2 17-05-2006 1,64 3 17-05-2006 1,64 17-05-2006 1,64 5 17-05-2006 1,65 6 17-05-2006 1,66 7 18 05 2006 1,63 8 18-05-2006 1,63 9 18-05-2006 1,63 IO 18-05-2006 1,65 II 18-05-2006 1,65 12 18-05-2006 1,65 13 19-05-2006 1,67 l' 19 05 2006 1,66 15 19-05-2006 1,66 16 19-05-2006 1,66 17 19 05 2006 1,65 18 19-05-2006 1,66 19 22-05-2006 1,67 20 22-05-2006 1,67 21 22-05-2006 1,70 22 22 05 2006 1,70 23 23 05-2006 1,66 2. 23-05-2006 1,66 25 23-05-2006 1,69 26 23 05-2006 1,66 27 23 05-2006 1,66 28 23 05-2006 1,67 29 23 05-2006 1,69 30 Middel 1,66 sd 0,019 CV/% 1,15 Bilag 7

~==:.o-..-, =::t~_::::::. IREFI15196 r6i ORIGINAL Nova 8 Calibrator Pack Kallbralorpackung. nakita Bo9IJovotJl11T). Paquela del cahbrador. Coffrel elaioll, Con/ezlone callbratore. f \" 'J, (.,-9 J\ 'l 'J, Pacole de cahbrador. Kahbralorpakel ICAL[jU >l000ml ICALIID >660ml ICALlc I ~60mL ISOLNl[] "'25OmL [jjj l~t~ffinerlwl ph 7.384 @ 37.0 C ph 6.84 @ 37.0 C ph 7.384 @ 37.0 C KCI 2 mol/l Mg" 0.50 mmol/l Mg" 0.50 mmol/l Mg" 1.50 mmolll Na' 140.0 mmolll Na' 75.0 mmol/l Na' 140.0 mmolll K' 4.00 mmol/l K' 10.0 mmol/l K' 4.00 mmoul Ca" 1.00 mmol/l Ca" 2,00 mmol/l Ca" 1.00 mmol/l Tho..... _... _...,...- -_... _'"", = 0).........::.."=.::.;0:-... :~t.::-.,~'t';... _.._._..._'.. UOOM... _...--. c ~... M, _.. _.. _ C'~'''''_ ~, 7Fi.!f'~~-.'"m'.._-_.. -._-...... <:0'...'... _.,- _..._,....-.... co.-... _ "'-----_...... ;:...:::-~I--...'q.-..., --... ".Co' ---~-_.~ ~'~_..._..~_......_...,.-..._.._.._...... 0_... -_ ="':.:':''::,':=.::':.:.O:=w~.''T'o~~ _. -~ -.,...-..._-"_.~.._..._-,~-, =--=''';:::'~-:==:=:"'~:-''.:;;:o...._._----.... -_.- _~,,,,-_. wo..:..._...._.._... _..---_...-..._._...- ~._.._----~- ~.,...-..._.. _.._-,--_.._,'- c.._.. _.._._...' _.. _..._-..._...-. -~_ '". _...~_T._. _ ~... ""...... ø --.."",.,-_... --' _...:_t:.. -...-.. ---- :=r-.=.~'u..=.~-"'-----... =r::=.;,....._"' '_.....,'-=----- ~..:_--"' ----.:.-:::::..:;;=.:.._-------_... n' t.. _.. ::~~"-:-~~:- ==::..."'"="::.~=...':'::-":r ø --... _0.-... ~..."".' '.. co...,,""ø ~_~.._ ~ _.- - _.--...'-_. -..._----~--<---- =--='--:':-:":=~~~':: _,, --- _..._~_.....--~_ --~--_.. ----,,_._-"'--- -"-I...--... ' -~...","_._---~-,....._ --~---_.. _.- r _ --~- ~=-==-_..._...._..., 1--"--'-'.-.-..,,, -1"...'''--,-_ -......_,.. "'..._-_..'..._--.,...,...~,....._-' _...,_ ---,...". """.m "'''':<:...'..T''_..-...-._,-- -,""'-' ~;r.:~::."':-"'='=~=::.:o,:~o;:.'":=...< =::=:'::'---,..._,..._..., -- - c.o-, _r~_,..._.._..._-_..._---_., ~_...-...-- -.-...- -,_.....ø... -_._.- ------_ -'"'=:..'"=='~~'L':"'~~~._-~... ø_,....._..... =--.:==:':. ''. ---..._~_._~-- ----._--- :.::r-'...;-_...- _ -.-...-, -e-.-."..."..._._~.... _._ r... --'-,... I EC IREP INova Biomedicai UK C3-5, Evans Business Centre. Deeside Induslrial Deeside. Flintshire. UK. CH5 2JZ Park, Waltham, MA 02454-9141 U.StA. LPN 38.1'" R..., wc

Klinisk biokemisk afdeling Amtssygehuset i Gentofte 02-03-2007 PræcisioDsprofil Analyse: P- Calcillm-ion(frit)jstoJk. Målt på: Nova Il i mmolll. Prøve I Prøve 2 Prøve 3 Prøve 4 Prøve 5 l. hest 0,59 1,20 2, Il 2,44 1,52 2. best 0.59 1,19 2,10 2,44 1,52 3. hest 0,59 1,19 2,11 2,44 1,52 4. best 0.59 1,20 2,10 2,43 1,52 5. hest 0,58 1,20 2,11 2,43 1.52 6. hest 0.59 1,20 2,08 2,44 1,54 7. hest 0.59 1,20 2.09 2,43 1,54 8. hest 0,59 1,20 2,08 2,44 1.52 9. hest 0,59 1,20 2,08 2,45 1,54 IO. hest 0.59 1,19 2,08 2,44 1,54 Middel 0.59 1,20 2,09 2,44 1,54 CV/% 0.54 0.40 0.64 0.26 0.67 Referenceinterval Nedre grænse 1,15 Øvre grænse 1.35 0,80,------------------------------., 0,70 +----------------- I 0,60 t--------=:::;:;~--j_-...;;;;;;:::::::--------- j x~ ~ 0,50 +------c //"--------+--+---------'...~:c-------1 "$ ""'- 80,40 +----------1-><---1----------------'...---1 0,30 +------ 0,20 +----------+--+-------------------1 0,10 +---------+------1----1------------------j x x x 0,00 +- ~---~--'---~---.J---.J-_--~--_---~--~-----' 0,50 0,75 1,00 1,25 1,50 1,75 2,00 mmolfl I--Refcrenccintcrval -Poly. (Profil) I 2,25 2,50 Bilag 9

Klinisk biokemisk afdeling Gentofte Hospital 21-06-2007 Præcisionsprofil Analyse: P- ioniseret Calcium (PH7,4) Målt på: Nova 8. ; mmol/l Prøve 1 Prøve 2 Prøve 3 Prøve 4 Prøve 5 I. best 0,53 1,07 2,11 2,54 1,78 2. hest 0,53 1,07 2,11 2,54 1,78 3. hest 0.53 1,07 2,12 2,55 1,79 4. hest 0,53 1,07 2,12 2,54 1,81 5. best 0,52 1,08 2,12 2,57 1,76 6. bes! 0,53 1,08 2, [3 2,55 1,76 7. bes! 0,53 1,08 2.11 2,55 1,78 8. bcst 0,53 1,09 2,13 2,54 1,76 9. best 0,53 1,09 2.13 2.55 1,78 IO. best 0,53 1,09 2,13 2,55 1,78 Middel 0.53 1,08 2,121 2,548 1,78 CV/% 0.60 0.81 0,41 0.36 0.87 Referenceinterval Nedre gr.rnst: 1,15 Øvre grænse 1.35 1,00 0,80 ~O,60./ x ~ - > U 0,40 ~ '" 0,20 0,00,, 0,50 0,75 1,00 1,25 1,50 1,75 Koncentration 1-- Referenceinterval - Poly. (Profil)I 2,00 2,25 2,50 2,75 Bilag 10

Klin,s1< b,ot;em,s1< afdeling Gentofte Hospital. IS-06-1007 Linearitet nalyu: P-Cøkillm-iotl(fril);stoft. Milt pi: Nova8 Reagen$lot nr.: 15196 Kalibrator lot nr.: 15196 AccelIteret bias"!. :4 4 koncentrationsniveauer Lineariteu Linearites Målt kontrol kontrol koncentration middelværdi inten-al (y.akse) (x akse) mmovl mmou1 Pool I Lln3nlt1s 1<ont.ol l 0,39-0,69 0,54 0.56 Pool 2 Ilname151<ontroI2 0,96-1.26 l. 1l 1.17 Pool 3 hnamcts 1<ontrol 3 1,79-2,19 1.99 2.08 Pool 4 hnantets kontrol 4 2,07.2.57 2.32 2.42 3.00 Opgivet middelværdi koncentration! Målt koncentration (mmol/i) ~ 2,50 E E 2.00 --.0.. 1.50 1.00 ~ "~ 0,50 ~ ~ ~ ~ 0.00 o 0.5 I 1.5 2 2.S 3 Opgi\'et middelværdi koncentration mmolll. Bilag II.

DISPLAY RANGES FOR ANAlYTES Display Ranges for Analytes The foliowing display ranges are used by the Nova anajyzer to check for result range errors (low ar high). lithe cancentration of an analyte falls outside the range for a specimen sample type, the speci.men cannot be tested for that analyte. Table 1 5. Display Range by Sample Type Analytes SerumlPlasmalWhole Blood Somum Potassium lonized Calcium Ionized Magnesium ph Hct '" 50 220 mmolll 0.8-11.0 mmoljl 0.1-5.5 mmolll 0.1-3.0 mmolll 6.5-8.0 12-70% '" Whole Blood oniy. When the Unconditional Reporting feature is On (See Introduction to Major Software Functions, Diagnostics), the instrument will display all test results, regardless af error conditions. The accuracy of test results which exceed the display ranges of the instrument cannot be assured. 1. Tietz, Norbert W., ed. 1986. Textbook ofclinical Chemistry. W.B. Saunders Co_ 2. Slatland, Bernard. 1987. Clinical Decision levels for lab Tests. Medicai Economics Books. 3. This is an average reference range thai was delermined from reference ranges established in a dozen or so reporting institutions that have been working with Nova analyzers. The exlremes of lhe reference ranges in these institutions were 0.43-0.57 to 0.46-0.62 with most being 0.45 Q.60 mmovl. Updale to Rev. H 7/98 1 35

Bi \Clj \3 _ NOVA 8 REFERENCE MANUAL Acceptable Anticoagulants Sodium and lithium heparin are the recommended anticoagulants for plasma specimens. Zinc heparin, EDTA, citrate, oxalate, ar sodium fluoride are not recommended for use as anticoagulants. Depending an the amount afbeparin used in the collection container and whether it is filled to capacity witb blood, heparin concentr-atiods of20 LU. per ml Dr more may result. 80dium beparin may elevate sodium results, and lithium heparin will elevate lithium results. Liquid heparin when present in excess may cause &rors by dilution. Our experience suggests that lyophilized sodium or lithium beparin giving a final concentration in blood ofnotmore than 30 LU. per ml is acceptable in the critical care laboratory. Nova Analyzer users should take careful note afthese considerations when establishing reference intervals and interpreting results. Interfering Substances At a levelof 1 romalil Mg+'", a level af5 mmolll Phosphate decreased the observed M~ by 20%. Reference Intervals Each laboratory should establish and maintain its own reference intervals based on its patient population. Table 1-4. Reference Intervals Sodium l PotaBBium l IonizedCalcium 2 Ioni2edM.gnesium 3 Ret (Male)). CFema1e) Seru.n 136-146 mdloljl 3.6-5.0 mdloljl 1.13-1.32 mmol/l 0.45.{).60 mmol/l WholeBlood 1.13-1.32 mmolll 0.45-0.60 mmolll 43 51% 38-46% 1-34 Update to Rev. H 7/98

t':>i \c:9 J'I _ ONE-POINT CAlIBRATION 7. The electrodes take Standard A millivolt readings for Na~, K', ph, ica, img, and Hct. 8. Using the millivolt readings taken in steps 1-7, the anajyzer's microprocessor calculates the slopes for Na~, K", ph, ica, img, and Hct. One-point Calibration Electrode drift is a gradual change in electrode potential that can adversely affect electrode performance. The Nova 8 Analyzer performs a l-point calibration during every analysis to determine ifthe drift for any electtode falls autside its acceptable range. Detection af excessive drift generates the appropriate error eades, depending OD the analysis mode. The l-point calibration sequence has the following steps: 1. Standard A is pumped through the sensor block during analysis. The electrode takes Standard A millivolt readings for Na', K', ph, ica, and img. 2. For a stat analysis, the drift for each electrode is determined by comparing the Standard A readings taken in step 1 to the Standard A readings taken during the last 2-point calibration, 3. For a tray analysis, the drift for each electrode is determined by comparing the Standard A readings taken in step 1 to the Standard A readings taken during the last 2~point calibration and the last l-pomt calibration (the preceding tray analysis). One-Point Calibration Direct vs. Indirect Methodologies TIDs section compares direct and indirect methodologies for the measurement of analytes and explains how the Nova Belectrode's offset values were determined. Plasma Volume Displacemenl Error The small differences in sodium, potassium, ionized magnesium, and ionized calcium results obtained by direct potentiometry and indirect methods in norinal samples are caused by insoluble lipids and plasma 7 13

Gentofte Hospital Klinisk-biokemisk afdeling Bilag 15 Korrekthed bestemt v.bj.a. certificeret reference materiale!analyse: Reagens lot nr.: Målt på: Enhed: P-CøJiamHon(fri1), stojk. 15196 NoVII, mmovl Certificeret værdi (CCR.\I): 1,71 Ekspanderet usikkerhed på CRM 0,06 k-værdi 1 U ClI;\1 0.04 N,. Dato Værdi I 1,76 2 1,77 3 1,77 4 1.77 26-06-2006 5 1,77 6 1,77 7 1.76 8 1,76 CmiliD~ - Middelværdi af de 8 målinger 1,766 U milio2 - sd 0,01 2 * kombineret usikkerhed (u kombin...j = 2 * ~U~åling + U~RM 0,08\ Numerisk forskel på målt og oplyst værdi (.åmatorialo) = IC CRM - CMåling 0,056 Hvis 2 w U kombinmt > åmatrriale er der ingen forskel Resultatet er indenfor den certificerede værdi ± oplyst usikkerhed Bilag IS-17/BIAS(I,71)

Genlofte Hospital Klinisk biokemisk afdeling Bilag 16 Korrekthed bestemt v.bj.a. certificeret reference materiale IADal)'se:!ReageDs lot nr.: Milt pi: 1'-C~(friJ).u.jt. lsi", tilbed: Certifken"1 værdi (CCR-\4): IJ7 Ekspanden"t usikkerhed pi CRM 0.07. ~ ærd; 1 II ca.\! O.ØJ! N,. Dato Værdi l 1.41 2 1,42 3 1,42 4 1,42 26-06 2006 5 1,42 6 1,40 7 1,39 8 1,39 emilia!: - Middelværdi af de 8 malinger 1,409 "miliø -sd 0,01 2 * kombineret USikkerhed (u...mbinml) - 2'" 'u 2. + U 2 0,075 'V Måhng CRM Numerisk forskel pa malt og oplyst værdi (å"",lfrialj -IC CRM - CMaling 0,039 Hvis 2 "kombintnl > A...atrrialt er der ingen forskel Resultatet er indenfor den certificerede værdi ± oplyst usikkerhed Bilag 15 1718IAS(1.37)

Gentofte Hospital Klinisk-biokemisk afdeling Bilag 17 Korrekthed bestemt v.hj.a. certificeret reference materiale lanalyse:!reagens 101 nr.: Milt pi: Eohed: P-CiIJlCfl1rt-ilHr{frll), støft. 15116 N.H' Crrtifi('e~t værdi (CaL\I): 1,09 Ekspandent usikkerhed pi CRM 0,05 "'I'i~,di ] u CJl.\1 0,02.5 N,. Dato Værdi l \,13 2 1,13 ) 1,13 4 1,13 26-06-2006 5 1,12 6 1,12 7 1,11 8 1,10 C m1 1U:: = Middelværdi af de 8 målinger 1,121 u miliø - sd 0,01 2 * kom bine ret usikkerhed (u ko"'bin~..ei)::: 2 * lu 2. + U 2 0,055 -V Måling CRM Numerisk fol"$kel på målt og oplyst værdi (A...J::: IC CRM - CMålWlg 0,031 Hvis 2 U kombillt~t > Amattnalt er der ingen forskel Resultatet er indenfor den certificerede værdi ± oplyst usikkerhed Bilag 15 171DIAS (1,09)

Table 2. Combined Standard Uncertainties (mmolll) and Effective Oegrees of Freedom for Electrolytes ElecO'olyte Level 1 Level 2 Level 3 u, 'ou u, 'ou u, 'ou Total Calcium 0.0096 51 0.0073 158 0.0066 58 Lithium 0.014 13570 0.0086 261200 0.0035 333250 Magnesium O.OID 3678 0.0065 2713 0.0030 28512 Potassium 0.022 191 0.014 4118 0.0072 2534 Sodium 0.69 716 0.80 840 0.93 561 Table 3. Reference Concentrations (mmolll and mg/dl) for lonized Calcium in SRM 956b Level l Level 2 Level 3 tonized Calcium I mmolll mg/dl 1.71 ± 0.08 6.87 ± 0.33 1.37 ± 0.07 5.48 ± 0.28 1.09 4.38 ± 0.05 ± 0.21 The resulls are cxp!'cssed as Ihc reference valuc ± thc expandcd uncenainty. Tlte expanded unceltainty is expres.scd as a 95 % confidcncc interval. INSTRUCTIONS FOR USE FOR ALL ELECTROLYTES EXCEPT IONIZED CALCIUM Place the ampoule to be used inside another container. such as a plastic beaker. to ensure containmenl af the serum in case the ampoule cracks. Each ampoule shoujd be inspected carefully for circular cracks at the base. If the ampoulc is cracked. ar has visible deposits of serum materiaion the outside, it should not be used. The serum in intaet ampoules should be thawed to room tempcrature. and mixed by inverting gemly at least five times befare sampling. When opcning ampoules, wear appropriate eye proteetion. gloves and protective c1othing. Check. that all of the Iiquid has drained out of the neck of the ampoule. lf needed, gently tap the neck to facilitate drainage. Opcn the ampoulc by snapping off the top at the narrowest segment of the neck. Ampoules should not be resealed. Once opened. the comems of the ampouje should be used as saon as possible. Transfer the solution from the ampoule using a suitable transfer pipctte. DO NOT PIPElTE BY MOUTI-I. Pouring solution out af the ampoule is not recommended as the narrow cross seclion of the neck does not facilitate easy exchange of liquid and air. SPECIAL SAMPLE HANDLING INSTRUCTIONS FOR MEASUREMENT OF IONIZED CALCIUM Because of the influence af ph an ionized calcium, it is important that the samples arc thawed and re-equilibrated with the gas in the ampoule headspace using the spccified conditions given below [6]. 1. Remove samples from freezer and thaw at ambient temperature for 1 hour and 40 minutes. NOTE: ambienl temperature must be between 20 C to 24 Ge. 2. During the first few minutes of thawing, inspcct ampoules carefully for cracks or breaks. Arnpoules that are cracked ar broken shauld be discarded. 3. Aner the l hour and 40 minutes thawing period. shake each ampoule vigorously with an up and down motion along the cylindricai axis for 10 secands to create foam. 4. Wait an additional 30 minutes atter shaking. then begin analyzing the samples. 5. Open the ampoule and aspirate the sample from as dose as possibie to the bottom of the ampoule. The sample must be introduced inta the anajyzer within ane minute af opening af the ampoule. 6. If it is not possibie to aspirate sample direct1y from the ampoule into the analyzer for the particular system being used. the sample may be aspirated inta a syringe while minimizing contact with air. NOTE: The sample should be analyzed wilhin ane minute af opening the ampoule. SRM 956b Page 4 af 5

NOVA 8 REFERENCE MANUAL Acceptable Samples The following chart lists acceptable sample types fol" the different parameters measurerl: PARAMETER Sodium Potassium Ionized Calcium Ionized Magnesium pr Ret SPECJMEN MATRIX Serum, plasma, Ol" whole blood Serum, plasma, Ol" whole blood Serum, plasma, Ol" whole blood Serum, plasma, Ol" whole blood Serum, plasma, Ol" whole blood Whole blood only Sample Handling Requirements WARNING: Blood samples and blood products are potential * sources efhepatitis and ether infeetious agents. Handle au blood products and {lewpath eomponents with eare. Gloves and proteetive clothing are recommended. Proper sample handling is critical to ensure that values abtainerl aceu~ rately reflect the in viva state. Ensure that all samples have been abtained and stored fallowing consistent, clinically accepted protocols. The following is a discussion afgeneral guidelines in reference to sample collectionlhandling requirements for Nova B. The NCCLS Approved Guideline OD Procedures for the Handling and Processing afblood Specimens (Document H 1B-A Val. 10 No. 12) mdicates that serum Ol" plasma should be physical1y separated from contaet with cells as saon as possibie to a maximum time limit af2 hours from the time afcollection. It is also recommend that a contact time afless than 2 hours be established for serum Ol" plasma samples intenrled for potassium analysis. Serum Collect blaorl for serum samples in plain vacuum tubes Ol" serum separator tubes. Evacuated collection tubes should be filled completely to mjnjmize the 10ss afcarbon dioxide. Allow the sample to dot for ap proximately ane halfhour. Following clot retraction, centrifuge at 1000 RCF for 10 to 15 minutes, uncap the tube, and use a syringe/bulb pipette to obtain a serum sample. Take the sample from the area as close to the cells as possible. Ifthe sample cannot be analyzed immediately, the separated serum should remain at 22'C (room temperature) for DO longer than 8 hours. IT assays will nat be completed within 8 houts, the

ACCEPTABLE SAMPLES I serum sample should be stored refrigerated at 2 to 8 C. If assays will not he completed within 48 hours or lithe serum sample is to be stored beyond 48 hours, the samples are to be stored frozen at _20 C. Plasma CoHed whole blood for plasma samples with minimal stasis, without exercise of the arm, in vacuum tubes containing either sodium or lithium heparin. Whole blood specimens are not to be chilled unless there are documented recornmendations for doing so. Obtain plasma by centrifuging heparinized whole blood within ane hour afcollection. Following centrliugation at 1000 RCF for 10 to 15 minutes, remove the cap and use a syringe/bulb pipette to obtain a plasma sample. Take the sample from the area dose to the cells to obtain as anaerobic a sample as possible. Plasma samples more than ane hout old should he centri fuged immediately prior to analysis to remove any fibrin clots. Ifassays will not be completed within 8 hours, the plasma sample should be stored refrigerated at 2 to S'C. Irassays wil! not be completed within 48 hours ar lithe plasma sample is to be stored beyond 48 houts, the samples are to be stored frozen at -20 C. Whele Sleed Blood samples should be calleeted with minimal stasis, without exercise ofthe arm. Collect blood for analysis an the Nova 8 in V8cuum tubes containing either sodium or lithium heparin. Whole blood specimens are not to he chilled unless there are documented recorrunendations for doing so. It is particularly important to ensure that samples are well mixed before introduction into the analyzer. The NCeLS Approved Guideline an Procedures for the Handling and Processing afblood Specimens (Docwnent H 18-A Val. 10 No. 12) indicates that serum ar plasma should be physically separated from contact with cells as soon as possibie to a maximum time limit of2 houts from the time of collection. It is also recommended that a contact time ofless than 2 hours be established for serum ar plasma samples intended for potassium analysis. References: NeeLS. Procedures for the Handling and Processing ofblood. Dacument HIB-A ValID No. 12. NCCLS. Procedures for the Collection ofdiagnostic Blood Specimen..~ by Venipuncture. Document H3 A3 VollD No. 12. Jacobs., Kasten, DeMott, and Walfson, ed. 1990. Laboratory Test Handbook. Lexi-Comp Ine. Tietz, N.W., ed. 1986. Textbook ofclinical Chemistry. W.B. Saunders ~. 1~3

Gentofte Hospital Klinisk-biokemisk afdeling Ekspanderet relativ kombineret standard måleusikkerhed j:':,ar NPU 01446 P-Calcium-ion(frit);stofk målt på NOVA 8 Bilag 20 Ingen Biologisk variationi) (CV w ) / % oolvsninl?er, "O Bidrag til måleusikkerhed '" ~.= '" "'~ -"-,, '" Analytisk (CViolennediær imnræcision) I % 1,34 -< = -" o;; Kalibrator 2 ) (CVkalibralor) / % 1,0 = Præanalytisk') (CVpræanalytisk) / % Relativ kombineret standardusikkerhed I 0/0 Ingen oolysninger ~2'" (CV~"tam«b:f,m~lC<...,n + CV :oiotr..'" +CV ~""">lylo.k) 2,4 Dc relative usikkerhedsbidrag - niveau Ca++ = 0,82 0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 ),6 Relativ kombineret standardusikkerhed I % O Kalibrator Analytisk I Referencer: I) Validitetserklæring V-046/02 2) Brev af 15.06.2006 fra Nova 310medieal 02-03-2007 Akt Ca-NOVA8 Side 1 af 1

Ekspanderet relativ kombineret standard måleusikkerhed Klinisk-biokemisk afdeling ~,or NPU 01446 P-Calcium-ion(frit);stofk målt på NOVA 8 Gentofte Hospital Bilag 21,." Biologisk variation l ) (CVw) / % Ingen oplysninger Bidrag til måleusikkerhed ~..c " >. ~ - " Analytisk (CVintennediær impra::cision) / % 1,03,,"" ="" <.~ Kalibrator 2 ) (CVkalibralor) / % 1,0 = Præanalytisk 3 Ingen ) (CVpræallal)1isl.J / % oo[vsnin!!er Relativ kombineret.i?*(cv' +C' +C' ) 2,0 standardusikkerhed / % - "'~',mpt~<""". V hl_or V Pfll:on"~t'" De relative usikkerhedsbidrag - niveau Ca++ = 1,45 1,0 1,0 1,0 1,0 1,0 1,0 1,0 1,0 1,0 1,0 1,0 Relativ kombineret standardusikkerhed I % O Kalibrator Analytisk I Referencer: I) Validiletserklæring V-046/02 2) Brev af 15.06.2006 fra Nova Biomedical 02-03-2007 Akt Ca-NOVA8 Side l af l

Ekspanderet relativ kombineret standard måleusikkerhed Klinisk biokemisk afdeling C:,or NPU 04144 P-Calcium-ion(frit);stofk (ph=7,4) målt på NOVA 8 Gentofte Hospital Bilag 22 "O,~ ~ Biologisk variation l ) (CV w ) / % Ingen oplysnin2er Bidrag til måleusikkerhed ~.c ~~ - ~.:< ~ Analytisk (CVintermediær imdrædsion) / % 0,93 C.:<.~ -<: Kalibrator 2 ) (CVkalibmtor) / % 1,0 = Præanalytisk 3 ) (CVpræanalYlisk) I % Relativ kombineret standardusikkerhed I % Ingen oplysninger i2*(cv:,«mc.lo"''''''''''<'''''''" +CV:...", +CV;,,,...Ylol;) 1,9 Dc relative usikkerhedsbidrag - niveau Ca++(pH=7,4) = 0,97 0,9 0,9 0,9 0,9 1,0 1,0 1,0 1,0 R~lativ kombineret standardusikkerhed I % O Kalibrator Analytisk I Referencer: l) Validitetserklæring V-046/02 2) Brev af 15.06.2006 fra Nova Biomedicai 02-03-2007 Ca 7.4 -NOVA8 Side l afl

Gentofte Hospital Klinisk-biokemisk afdeling Ekspanderet relativ kombineret standard måleusikkerhed for NPU 04144 P-Calcium-ion(frit);stofk (ph=7,4) målt på NOVA S Bilag 23, "O Biologisk variation I) (CV w ) / % Ingen oolvsninller Bidrag til måleusikkerhed " ~.c "'~ -,,-"" Analytisk (CVi01ennediær imoræcision) / % 1,15 =-" <r:.- Kalibrator 2 ~ ) = Præanalytisk 3 ) (CVpræanalytisk) / % (CVkalibrator) / % 1,0 Relativ kombineret standardusikkerhed / % '"'enn«i.,"ntr'''c'''''" + V kal,!>'..", + p,.,...:oiy,m Ingen oplysninger l/z'(cv' C' CV' ) 2,2 De relative usikkerhedsbidrag - niveau Ca++ (ph=7,4)= 1,66 0,9 1,0 1,0 I,I I,I 1,2 1,2 Relativ kombineret standardusikkerhed I %:- D Kalibrntor Analytisk I Referencer: l) Validitetserklæring V-046/02 2) Brev af 15.06.2006 fra Nova BiomedicaI 02-03-2007 Ca 7.4 -NOVA8 Side l af l

--, s~"

,. e s- System Komponent, lwantitetsart Enhed Refintervol al-antitrypsin, massek. gjl 0,97-1,68 PP 1.29-2.23 s- al-antitrypsin, stotk. IJmolj! 18-41 (KAS) s- Aspartat-aminotransferase, enzk. U/I k: s 35 (KASGl: s 50) m: s 50.,- Base (H+-bindingsgruppe), stofk.diff. (aktuel- norm) mmoljl -3,0 - +3,0 s- Basisk phosphatase. enzk. U/I 80-275 s- Bilirubiner, stof!<. ljmoljl 4-22 (KASGI; 4-17) s- Bilirubin (konjungeret), stolle IJmoljl s- C-reaktivt protein, massek. mg/l S" 10 [2J,- Calcium, stofk. mmoljl 2,20-2,60,- Calcium-ion (fri), stotk.(ph-7a) mmoljl 1,15-1,35,- Calcium, albumin korrigeret, stofk. mmol/l 2,14-2,52 du- Calcium, stofm. mmol 25-8,1 s- Carbamid, stof\:. mmoljl 2,5 7,5 U- (arbamid. stofle mmoljl 180-600 4 du- Carbamid, stofm. mmol 260-500 System Komponent Alder Enhed Ref.interval (Kilde) S Bilirubiner 1md-14år ~mol/i <2 (konjugeret) B S C-reaktivt protein lmd-14år mg/l <8 5 S C-reaktivt protein 1md-14år nmol/l <78 5 S Calcium 0-12mdr mmolll 2,10-2,62 1-14 år 2,17-2,66 S Calcium-ion (fri) 0-6mdr mmol/l 0,95-1,50 (ph:=7,4) 7mdr.-14år 1,22-1,37 S Carbamid 0-3mdr mmol/l 0,7-5,7 4-12mdr 0,4-5,0 1-14 år 1,8-7,5 n