Procedure:

ISE
Electrolytes: Na , K+, Cl-
+

Thisprocedureisvalidforthefollowingchemistryanalyzers:

AU400/AU400e AU640/AU640e
AU480 AU680
AU600 AU2700
AU5400 AU5800

Prepared By Date Adopted Supersedes Procedure #

Review Date Revision Date Signature

# of # of
Distributed to Copies Distributed to Copies

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

PRINCIPLE:
Thedeterminationofelectrolytes(sodium,potassium,andchloride)isoneof
themostimportantfunctionsintheclinicallaboratory.

Electrolytesaffectmostmetabolicprocesses.Theyservetomaintainosmotic
pressureandhydrationofvariousbodyfluidcompartments,properbodypH,
andregulationofappropriateheartandmusclefunctions. Electrolytesare
also involved in oxidationreduction reactions and participate as essential
partsorcofactorsinenzymereactions.1

INTENDED USE:
SystemreagentforthequantitativedeterminationofSodium,Potassium,
andChlorideconcentrationsinhumanserum,plasma,orurineonBeckman
CoulterAUClinicalChemistryanalyzers.

METHODOLOGY:
TheBeckmanCoulterAUSystemISEmoduleforNa+,K+,andClemploys
crownethermembraneelectrodesforsodiumandpotassium;andamolecular
orientedPVCmembraneforchloridethatarespecificforeachionofinterest
inthesample.AnelectricalpotentialisdevelopedaccordingtotheNernst
Equationforaspecificion.WhencomparedtotheInternalReference
Solution,thiselectricalpotentialistranslatedintovoltageandthenintothe
ionconcentrationofthesample.1

Methodsofdeterminingelectrolytesincludeemissionsspectrophotometry,
flamespectrophotometry,neutronactivationanalysis,atomicabsorption
spectroscopy,andionselectiveelectrodes.

SPECIMEN:
PATIENT PREPARATION:
Nonerequired.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Additionalinstructionsforpatientpreparationasdesignatedbythislaboratory:

TYPE:
Serumorplasma,freefromhemolysis,istherecommendedspecimen.
Separateserumfromredbloodcellsassoonaspossible.Ifplasmamustbe
used,therecommendedanticoagulantsarelithiumheparinand
ammoniumheparin.Avoidhemolysis,whichmayfalselyelevate
potassiumvalues.

Urinesamplesshouldbecollectedinaclean,leakproofcontainerand
shouldnotbeacidified.Iftransportisdelayed,specimensshouldbekept
refrigeratedat28C.

A24hourcollection2istherecommendedspecimentodeterminetheurine
sodium,potassiumorchloride.
Additionaltypeconditionsasdesignatedbythislaboratory:

HANDLING CONDITIONS:
Usefreshsampleforanalysiswhenpossible.Sodiumandpotassiumare
stableinserumforatleastoneweekwhenstoredat28C.Chlorideis
stableinserumforoneweekwhenstored230C.Storethesamplesina
stopperedtubeifanalysisisdelayed.2

Urinespecimensshouldbestoredat28C.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Additionalhandlingconditionsasdesignatedbythislaboratory:

EQUIPMENT AND MATERIALS:

EQUIPMENT:

BeckmanCoulterAU400/AU400e,AU480,AU600,AU640/AU640e,
AU680,AU2700,AU5400andAU5800analyzers.

MATERIALS:
Na+Electrode: CatNo.MU9194

K+Electrode: CatNo.MU9195

ClElectrode: CatNo.MU9196

ReferenceElectrode: CatNo.MU9197

ReactiveComponents:
ISE Buffer (AUH1011) ISE Reference (AUH1013)
Triethanolamine 0.1 mol/L Potassium Chloride 1.0 mol/L
Preservatives Preservatives

ISE Mid Standard Internal Reference

(AUH1012) Solution (AUH1017)
Sodium 4.3 mmol/L Potassium Chloride 3.3 mol/L
Potassium 0.13 mmol/L Silver Chloride Saturated
Chloride 3.1 mmol/L Preservatives
Preservatives

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

ISE Low Serum Standard ISE High Serum Standard

(AUH1014) (AUH1015)
Sodium 130 mmol/L Sodium 160 mmol/L
Potassium 3.5 mmol/L Potassium 6.0 mmol/L
Chloride 85 mmol/L Chloride 120 mmol/L
Preservatives Preservatives

Note: Urine standards are packages as a set (AU1016)

ISE Low Urine Standard ISE High Urine Standard
Sodium 50 mmol/L Sodium 200 mmol/L
Potassium 10 mmol/L Potassium 100 mmol/L
Chloride 50 mmol/L Chloride 180 mmol/L
Preservatives Preservatives

Note: Selectivity solutions are packages as a set (AUH1018)

Na+ Selectivity Check K+ Selectivity Check
Solution Solution
Sodium 150 mmol/L Potassium 5.0 mmol/L
Preservatives Preservatives

Reagentstoragelocationinthislaboratory:

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Testtubes1216mmindiameterorsamplecups (CatNo.AU1063).
Storagelocationoftesttubesorsamplecupsinthislaboratory:

ISE Low Serum Standard (Cat No. AUH1014)

ISE High Serum Standard (Cat No. AUH1015)
ISE Low/High Urine Standard (Cat No. AUH1016)

StoragelocationoftheChemistryCalibratorinthislaboratory:

Precautions:

1. TheISEReagentsandStandardsareforinvitrodiagnosticuse.

2. Donotingest.Harmfulifswallowed.

PREPARATION:
TheBeckmanCoulterAUSystemISEreagentsareliquid,readyforuse.
Nopreparationisneeded.ISEelectrodescomepreparedandreadyto
installonboardtheanalyzer.

Storage Requirements:
1. Theunopenedreagentsandstandardsarestableuntilthe
expirationdateprintedonthelabelwhenstoredat225Cwith
theexceptionoftheInternalReferenceSolution(CatNo.
AUH1017),whichisstored1525C.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

3. ISEBuffer(CatNo.AUH1011),ISEMidStandard(CatNo.
AUH1012),andISEReferenceSolution(CatNo.AUH1013)are
stablefor90dayswhenopenedandstoredintheISEreagent
compartmentoftheanalyzer.

4. Afteropening,ISELowandHighSerumStandards(CatNo.
AUH1014andAUH1015),ISEUrineLow/HighStandard(CatNo.
AUH1016),andNa/KSelectivityCheckSolutions(CatNo.
AUH1018)maybestoredat225Cforupto90daysprovidedthe
capisreplacedimmediatelyafteruse.ISEInternalReference
Solution(CatNo.AUH1017)maybestoredat1525Cforupto90
daysprovidedthecapisreplacedimmediatelyafteruse.

Indications of Deterioration:
Discolorationofthereagent,visiblesignsofmicrobialgrowth,
turbidityorprecipitationinreagentmayindicatedegradationand
warrantdiscontinuanceofuse.
Additionalstoragerequirementsasdesignatedbythislaboratory:

PERFORMANCE PARAMETERS:
ThefollowingdatawasobtainedusingtheISEReagentsBeckmanCoulter
AUanalyzersaccordingtoestablishedprocedures.Resultsobtainedat
individualfacilitiesmaydiffer.

PRECISION5:
Estimatesofprecision,basedonCLSIrecommendations4,areconsistent
withtypicalperformance.Thewithinrunprecisionislessthan3%CVand
totalprecisionislessthan5%CV.Assaysofcontrolseraandpooledurine
sampleswerecarriedoutanddatareducedfollowingCLSIguidelines.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

SERUM, N=100 Within run Total

Mean, mEq/L SD CV% SD CV%
Na+ Level 1 124.4 0.5 0.4 0.8 0.6
+
Na Level 2 143.0 0.5 0.3 0.8 0.5
K+ Level 1 2.9 0.2 0.6 0.02 0.7
K+ Level 2 5.7 0.04 0.8 0.05 0.9
Cl- Level 1 85.9 0.4 0.5 0.6 0.6
Cl- Level 2 112.3 0.5 0.5 0.6 0.6

URINE, N=100 Within run Total

Mean, mEq/L SD CV% SD CV%
+
Na Level 1 88 0.6 0.7 0.8 0.9
+
Na Level 2 182.3 0.7 0.4 1.5 0.8
K+ Level 1 32.1 0.3 0.8 0.3 1.0
K+ Level 2 85.8 0.5 0.6 0.9 1.1
Cl- Level 1 111.3 0.7 0.6 0.9 0.8
Cl- Level 2 238.8 0.9 0.4 1.4 0.6

METHOD COMPARISON5:
PatientsampleswereusedtocomparetheBeckmanCoulterAU
SystemISE.RepresentativeperformancedataonAUanalyzersis
showninthenexttable.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Serum:

Y Method AU640 / AU640e

X Method AU600
Na+ K+ Cl-
Slope 1.01 0.99 1.01
Intercept 0.3 0.12 0.7
Corr. Coeff. (r) 0.998 0.999 0.999
No. of Samples (n) 230 230 230

Urine:

Y Method AU640 / AU640e

X Method AU600
Na+ K+ Cl-
Slope 1.01 0.99 0.99
Intercept 1.5 4.7 -0.4
Corr. Coeff. (r) 0.999 0.999 0.999
No. of Samples (n) 184 184 184

CALIBRATION:
STANDARD PREPARATION:
PerformamultipointcalibrationbyusingtheautomatedISEcalibration
withtheappropriatestandardsplacedinthelabeledpositionsonthe
STATtableordesignatedISEStandardSolutionarea.Thefrequencyof
calibrationisdaily.CalibrationoftheISEmethodsisaccomplishedbythe
useoftheBeckmanCoulterISEstandardsforserumorurine.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

RefertotheISEsectionoftheappropriateAUUserGuideforacomplete
listingofcalibrationprocedures.

CALIBRATION PROCEDURE:
Recalibrationofthistestisrequiredwhenanyoftheseconditionsexist:

1. Areagentlotnumberhaschangedorthereisanobservedshiftin
controlvalues.

2. Majorpreventativemaintenancewasperformedontheanalyzer.

3. Acriticalpartwasreplaced.

QUALITY CONTROL:
DuringoperationoftheBeckmanCoulterAUanalyzeratleasttwolevelsof
anappropriatequalitycontrolmaterialshouldbetestedaminimumofoncea
day.Inaddition,controlsshouldbeperformedaftercalibration,witheach
newlotofreagents,andafterspecificmaintenanceortroubleshootingsteps
describedintheappropriateAUUsersGuide.Qualitycontroltestingshould
beperformedinaccordancewithregulatoryrequirementsandeach
laboratorysstandardprocedure.
Locationofcontrolsusedatthislaboratory.

ANALYZER PARAMETERS:
Acompletelistoftestparametersandoperatingprocedurescanbefoundin
theappropriateUsersGuideandatwww.beckmancoulter.com.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

CALCULATIONS:
ForSIUnitsmmol/LisequivalenttomEq/L.

REPORTING RESULTS:
REFERENCE RANGES:
Serum1: Urine1:

Na+:136145mEq/L Na+:40220mEq/day

K+:3.55.1mEq/L K+:25125mEq/day

Cl:98107mEq/L Cl:110250mEq/day

Expectedvaluesmayvarywithage,sex,dietandgeographicallocation.
Eachlaboratoryshoulddetermineitsownexpectedvaluesasdictatedby
goodlaboratorypractice.

Expectedreferencerangesinthislaboratory:

PROCEDURES FOR ABNORMAL RESULTS:

Abnormalresultsareflaggedbythelistedanalyzersaccordingtothe
normalvaluesenteredbytheuserintotheinstrumentparameters.

REPORTING FORMAT:
ResultsareautomaticallyprintedforeachsampleinmEq/Lat37C.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Additionalreportinginformationasdesignatedbythislaboratory:

LIMITATIONS:
TheBeckmanCoulterAUSystemISEproceduresarelinearasfollows:

Serum: Na+:50200mEq/L Urine: Na+:10400mEq/L

K+:1.010.0mEq/L K+:2.0200mEq/L

Cl:50200mEq/L Cl:15400mEq/L

Samplesexceedingthedynamicrangeoftheassayshouldbedilutedwith
deionizedwaterandreassayed.Theresultsobtainedmustbemultipliedby
thedilutionfactortoobtainthecorrectconcentrationfortheundiluted
sample.

INTERFERING SUBSTANCES:
Certainanticoagulants,preservatives,drugsandorganophiliccompounds
mayaffectelectrolytedeterminations.Forfurtherinformationon
interferingsubstances,refertoYoung3foracompilationofreported
interferenceswiththesetests.Visuallyturbidurinesamplesshouldbe
centrifugedpriortoanalysis2.

Grosslylipemicsamplesmayshowaninappropriatedecreaseinsodium,
potassium,andchlorideresultsduetovolumedisplacement.Such
samplesshouldbeultracentrifugedadtheanalysisperformedonthe
infranatant.(middleclearlayer).

TheinformationpresentedisbasedonresultsfromBeckmanCoulter
studiesandiscurrentatthedateofpublication.BeckmanCoulterInc.,
makesnorepresentationaboutthecompletenessoraccuracyofresults
generatedbyfuturestudies.

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 Procedure:
ISE
Electrolytes: Na , K+, Cl-
+

Laboratoryspecificprocedurenotes:

REFERENCES:
1. Tietz,N.W.editor.FundamentalsofClinicalChemistry.3 rdEdition,W.B.
Saunders,1987.

2. Pesce, A.J. Kaplan, L.A. editors, Methods in Clinical Chemistry. C.V.

MosbyCompany.1996.

3. Young, D.S.,EffectsofDrugs onClinicalLaboratory Tests,5 th Edition,

AACCPress.2000.

4. CLSI/NCCLSEvaluationProtocolEP5T2,1992.

5. DataonfileforspecificAUanalyzers.

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