ISE REAGENTS

AUH1011 Buffer 4 x 2000 mL

AUH1012 Mid-Standard 4 x 2000 mL
AUH1013 Reference 4 x 1000 mL
AUH1014 Low Serum Std. 4 x 100 mL
AUH1015 High Serum Std. 4 x 100 mL
AUH1016 High/Low Urine Std. 4 x 100 mL
AUH1017 Internal Ref. Sol. 2 x 25 mL
AUH1018 Na+/K+ Selectivity Check Sol. 2 x 25 mL
Intended Use
Reagent for the quantitative determination of Sodium, Potassium and Chloride concentrations in human serum and urine on the Beckman Coulter
ISE modules.

Summary
Electrolytes affect most metabolic processes. They serve to maintain osmotic pressure and hydration of various body fluid compartments, proper
body pH and regulation of appropriate heart and muscle functions. Electrolytes are also involved in oxidation-reduction reactions and participate as
1
essential parts or, cofactors, in enzyme reactions.

Methodology
The determination of electrolytes is one of the most important functions in the clinical laboratory. Methods of determining electrolytes include
emission spectrophotometry, flame spectrophotometry, neutron activation analysis, atomic absorption spectroscopy and ion selective electrodes.
+ + -
The ISE module for Na , K , and Cl employs crown ether membrane electrodes for sodium and potassium and a molecular oriented PVC membrane
for chloride that are specific for each ion of interest in the sample. An electrical potential is developed according to the Nernst Equation for a specific
ion. When compared to the Internal Reference Solution, this electrical potential is translated into voltage and then into the ion concentration of the
1
sample.

System Information
e e
For AU400/400 /480, AU600/640/640 /680 and AU2700/5400 Beckman Coulter Analyzers.

Contents, Reagents / Standards

Concentration of active ingredients:
ISE Low Serum Standard ISE Mid-Standard ISE High Serum Standard
+ + +
Na 130 mmol/L Na 4.3 mmol/L Na 160 mmol/L
+ + +
K 3.5 mmol/L K 0.13 mmol/L K 6 mmol/L
- - -
Cl 85 mmol/L Cl 3.1 mmol/L Cl 120 mmol/L
Preservatives Preservatives Preservatives
+
ISE Buffer ISE Reference ISE Na Selectivity Check
+
Triethanolamine 0.1 mol/L Potassium Chloride 1.00 mol/L Na 150 mmol/L
Preservatives Preservatives Preservatives
+
ISE Low/High Urine Standard ISE Internal Reference ISE K Selectivity Check
+ +
Na (Low) 50 mmol/L Potassium Chloride 3.3 mol/L K 5 mmol/L
(High) 200 mmol/L Silver Chloride Saturated Preservatives
+
K (Low) 10 mmol/L Preservatives
(High) 100 mmol/L
-
Cl (Low) 50 mmol/L
(High) 180 mmol/L
Preservatives

Precautions
1. For in vitro diagnostic use.
2. Do not ingest. Harmful if swallowed.

Preparation of Reagents
The reagents are ready to use. No preparation is required.

Storage and Stability

1. The unopened reagents are stable until the expiration date printed on the label when stored at 2 – 25°C. AUH1017 is stored at 15 - 25°C.
2. AUH1011, AUH1012, and AUH1013 are stable for 90 days when opened and stored in the ISE reagent compartment of the analyzer.
3. After opening AUH1014, AUH1015, AUH1016, and AUH1018 may be stored at 2 – 25°C for up to 90 days, provided the cap is replaced
immediately after each use. After opening, AUH1017 may be stored at 15 - 25°C for up to 90 days.

BAAUH1011.01 ISE
2009-08
ISE Reagents
Indications of Deterioration
Turbidity or precipitation in the unopened liquids and working reagents may indicate decomposition and warrant discontinuance of use. Refer to the
ISE section of the respective AU analyzer’s User’s Guide for additional information.

Specimen Collection And Preparation

Serum, free from hemolysis, is the recommended specimen.

Sample Stability
+
Separate serum from blood cells as soon as possible. Avoid hemolysis since it can lead to falsely elevated K values. If plasma must be used, the
recommended anticoagulants are lithium heparin and ammonium heparin. Urine samples should be collected in a clean, leak-proof container and
should not be acidified. If transport is delayed, specimens should be kept refrigerated at 2 - 8°C. The recommended collection times for urine
2
specimens, to determine the analyte of interest, are as follows.

Sodium: 24 hour collection

Potassium: 24 hour collection
Chloride: 24 hour collection

Sample Storage
Sodium and potassium are stable in serum for at least one week when stored at 2 - 8°C. Chloride is stable in serum for one week when stored at
2
2 - 30°C. Store the sample in a stoppered tube if analysis is delayed.
Urine specimens should be stored at 2 - 8°C.

Interfering Substances
Certain anticoagulants, preservatives, drugs, and organophilic compounds may affect electrolyte determinations. For further information on
3
interfering substances, refer to Young for a compilation of reported interferences with this test. Visually turbid urine specimens should be centrifuged
2
prior to analysis. Grossly lipemic samples may show an inappropriate decrease in sodium, potassium, and chloride results due to volume
displacement. Such samples should be ultracentrifuged and the analysis performed on the infranatant (middle clear layer).

Procedure
Materials Provided
ISE Buffer, Mid-Standard, Reference, High Serum Standard, Low Serum Standard, High/Low Urine Standard, Internal Reference Solution, and
+ +
Na /K Selectivity Check Solution.

Suggested Analytical Parameters

Refer to the ISE Section located in the respective AU analyzer’s User’s Guide for a complete listing of test parameters and procedure.

Stability of Final Reaction Mixture

The Beckman Coulter AU analyzer automatically computes every determination at the same interval at 37°C.

Calibration
Refer to the ISE section of the respective AU analyzer’s User’s Guide for a complete listing of the recommended calibration procedures. This
procedure should be used to calibrate daily.

Quality Control
During operation of the Beckman Coulter AU analyzer at least two levels of an appropriate control material should be tested a minimum of once a
day. In addition, controls should be performed after calibration, with each new lot of reagent, and after specific maintenance or troubleshooting steps
described in the appropriate Beckman Coulter AU analyzer User’s Guide. Quality control testing should be performed in accordance with regulatory
requirements and each laboratory’s standard procedure.
Appropriate qualified urine controls should be used during urine analysis.

Results
Results are automatically printed out for each sample in mEq/L.

Dynamic Range
The ISE Module procedures are linear in serum, plasma or urine samples as follows:
Serum Urine
+ +
Na 50 – 200 mEq/L Na 10 – 400 mEq/L
+ +
K 1.0 – 10.0 mEq/L K 2.0 – 200.0 mEq/L
- -
Cl 50 – 200 mEq/L Cl 15 – 400 mEq/L

Samples exceeding the dynamic range of the assay should be diluted with deionized water and reassayed. The results obtained must be multiplied
by the dilution factor to obtain the correct concentration for the undiluted sample.

Expected values1
+ + -
Serum: Na 136 - 145 mEq/L K 3.5 - 5.1 mEq/L Cl 98 - 107 mEq/L
+ + -
Urine: Na 40 - 220 mEq/day K 25 - 125 mEq/day Cl 110 - 250 mEq/day

Expected values may vary with age, sex, diet and geographical location. Good laboratory practice dictates that each laboratory determine its own
expected values.

ISE BAAUH1011.01
2009-08
 ISE Reagents
Specific Performance Characteristics
The following data was obtained using the ISE reagents on AU analyzer’s ISE modules according to established procedures. Results obtained in
individual laboratories may differ.

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Precision
4
Estimates of precision, based on CLSI recommendations , are consistent with typical performance. The within run precision is less than 3% CV and
total precision is less than 5% CV. Assays of control sera and pooled urine samples were carried out and the data reduced following the CLSI
guidelines referenced above.

Urine
N = 100 Within run
Mean SD CV%
+ + - + + - + + -
Na K Cl Na K Cl Na K Cl
Level 1 88 32.1 111.3 0.6 0.3 0.7 0.7 0.8 0.6
Level 2 182.3 85.8 238.8 0.7 0.5 0.9 0.4 0.6 0.4

N = 100 Total
Mean SD CV%
+ + - + + - + + -
Na K Cl Na K Cl Na K Cl
Level 1 88 32.1 111.3 0.8 0.3 0.9 0.9 1.0 0.8
Level 2 182.3 85.8 238.8 1.5 0.9 1.4 0.8 1.1 0.6

Serum
N = 100 Within run
Mean SD CV%
+ + - + + - + + -
Na K Cl Na K Cl Na K Cl
Level 1 124.4 2.9 85.9 0.5 0.2 0.4 0.4 0.6 0.5
Level 2 143 5.7 112.3 0.5 0.04 0.5 0.3 0.8 0.5

N = 100 Total
Mean SD CV%
+ + - + + - + + -
Na K Cl Na K Cl Na K Cl
Level 1 124.4 2.9 85.9 0.8 0.02 0.6 0.6 0.7 0.6
Level 2 143 5.7 112.3 0.8 0.05 0.6 0.5 0.9 0.6

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Method Comparison
A comparison of the Beckman Coulter AU Analyzers ISE was conducted with patient serum samples. The table below demonstrates representative
performance on AU analyzers.

Serum Comparison
e
AU640/640 vs AU600
+ + -
Na K Cl
N 230 230 230
r 0.998 0.999 0.999
slope 1.01 0.99 1.01
intercept 0.3 0.12 0.7

Urine Comparison
Beckman Coulter comparisons were conducted utilizing urine patient samples. The table below demonstrates representative performance on AU
analyzers.
e
AU640/640 /680 vs AU600
+ + -
Na K Cl
N 184 184 184
r 0.999 0.999 0.999
Slope 1.01 0.99 0.99
Intercept 1.5 4.7 -0.4

References
1. Tietz, N.W., editor, Fundamentals of Clinical Chemistry, 3rd Edition, W.B.Saunders 1987.
2. Pesce, A.J., Kaplan, L.A., editors, Methods in Clinical Chemistry, C.V. Mosby Company, 1996.
3. Young, D.S., Effects of Drugs on Clinical Laboratory Tests, 5th edition, AACC Press, 2000.
4. CLSI/NCCLS Evaluation Protocol EP5-T2, 1992.
5. Data is on file for specific AU analyzers.

Manufactured by: Beckman Coulter, Inc., 250 S. Kraemer Blvd. Brea, CA 92821, USA

BAAUH1011.01 ISE
2009-08