0003289923190COINV7.

CO2-L
Bicarbonate liquid Substrates
Order information
Analyzer(s) on which cobasc pack(s) can be used
COBASINTEGRA 400 plus
03289923 190 Bicarbonate liquid (250 tests) System-ID 0767255
COBAS INTEGRA 800
20751995 190 Ammonia/Ethanol/CO2 Calibrator (2 4 mL) System-ID 0751995
20752401 190 Ammonia/Ethanol/CO2 Control Normal (5 4 mL) System-ID 0752401
20753009 190 Ammonia/Ethanol/CO2 Control Abnormal (5 4 mL) System-ID 0753009
12149435 122 Precinorm U plus (10 3 mL) System-ID 0779997
12149435 160 Precinorm U plus (10 3 mL, for USA) System-ID 0779997
12149443 122 Precipath U plus (10 3 mL) System-ID 0780006
12149443 160 Precipath U plus (10 3 mL, for USA) System-ID 0780006

English Reagent handling

System information Ready for use
Test CO2L, testID 0625 Storage and stability
Intended use Shelf life at 28C See expiration date on
In vitro test for the quantitative determination of the bicarbonate (HCO3-) cobasc pack label
concentration in human serum and plasma on COBASINTEGRA systems.
COBASINTEGRA400 plus system
Summary
Bicarbonate is the second largest fraction of the anions in plasma. Included On-board in use at 1015C 6weeks
in this fraction are the bicarbonate (HCO3-) and carbonate (CO32-) ions, as COBASINTEGRA800 system
well as the carbamino compounds. At the physiological pH of blood, the
concentration of carbonate is 1/1000 that of bicarbonate. The carbamino On-board in use at 8C 6weeks
compounds are also present in such low quantities that they are generally Specimen collection and preparation
not mentioned specifically.
For specimen collection and preparation only use suitable tubes or
Several different methods for the determination of bicarbonate in serum and collection containers.
plasma have been reported. Most of these procedures utilize acidification of
the sample and conversion of all carbon dioxide forms to CO2 gas.1 The Only the specimens listed below were tested and found acceptable.
amount of gas formed is measured by manometric or volumetric devices, Serum
ion selective electrodes, or spectrophotometric techniques.2,3 These Plasma: Heparin (Li-, Na-, NH4+-) plasma
methods are either cumbersome, timeconsuming, techniqueoriented, The preferred specimen is from venous blood collected anaerobically in the
and/or require special equipment. usual manner for bicarbonate analysis. Bicarbonate content in uncapped
Enzymatic procedures using phosphoenolpyruvate carboxylase (PEPC) tubes decreases approximately 4mmol/L after one hour.6 It has been
have been described.4,5 reported that alkalinized serum stored in open cups is stable for up to
4hours.6
The bicarbonate content of serum or plasma is a significant indicator of
electrolyte dispersion and anion deficit. Together with pH determination, The sample types listed were tested with a selection of sample collection
bicarbonate measurements are used in the diagnosis and treatment of tubes that were commercially available at the time of testing, i.e. not all
numerous potentially serious disorders associated with acidbase available tubes of all manufacturers were tested. Sample collection systems
imbalance in the respiratory and metabolic systems. from various manufacturers may contain differing materials which could
affect the test results in some cases. When processing samples in primary
Test principle tubes (sample collection systems), follow the instructions of the tube
Bicarbonate reacts with phosphoenolpyruvate (PEP) in the presence of manufacturer.
PEPC to produce oxaloacetate and phosphate: Centrifuge samples containing precipitates before performing the assay.
Separate from erythrocytes and store tightly stoppered.
PEPC

PEP + HCO3- oxaloacetate + H2PO4- Stability: 7days at 48C7

The above reaction is coupled with one involving the transfer of a hydrogen 40hours at 1525C8,9
ion from NADH analog to oxaloacetate using MDH.
Storage of serum at 20C or 80C for up to 6months had no significant
effect.10
MDH

Oxaloacetate + NADH analog + H+ malate + NAD+ analog Materials provided

See Reagents working solutions section for reagents.
The resultant consumption of NADH analog causes a decrease in
absorbance at 409nm, which is proportional to the concentration of Assay
bicarbonate in the sample being assayed. For optimum performance of the assay follow the directions given in this
document for the analyzer concerned. Refer to the appropriate operators
Reagents - working solutions manual for analyzerspecific assay instructions.
R Phosphoenolpyruvate: 40mmol/L; NADH analog: 2mmol/L; Application for serum and plasma
MDH(porcine): 314.3kat/L; PEPC (microbial): 30.8kat/L
COBASINTEGRA400plus test definition
R is in position B.
Measuring mode Absorbance
Precautions and warnings
Pay attention to all precautions and warnings listed in Abs. calculation mode Endpoint
Section1/Introduction of this Method Manual. Reaction mode R-S
For USA: For prescription use only. Reaction direction Decrease

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0003289923190COINV7.0

CO2-L
Bicarbonate liquid Substrates

Wavelength A/B 409/512 nm Conversion factor: mmol/L 1 = mEq/L11

Calc. first/last 21/45 Limitations - interference
Unit mmol/L Criterion: Recovery within 10% of initial value.
Icterus:12 No significant interference up to an Iindex of 60 for conjugated
Pipetting parameters and unconjugated bilirubin (approximate conjugated and unconjugated
bilirubin concentration: 1026mol/L or 60mg/dL).
Diluent (H2O)
Hemolysis:12 No significant interference up to an Hindex of 1000
R 50 L 120 L (approximate hemoglobin concentration: 621mol/L or 1000mg/dL).
Sample 2 L 10 L Lipemia (Intralipid):12 No significant interference up to an Lindex of 2000.
There is poor correlation between the Lindex (corresponds to turbidity) and
Total volume 182 L triglycerides concentration.
Drugs: No interference was found at therapeutic concentrations using
common drug panels.13,14
COBASINTEGRA800 test definition
In very rare cases, gammopathy, in particular type IgM (Waldenstrms
Measuring mode Absorbance macroglobulinemia), may cause unreliable results.15
Abs. calculation mode Endpoint An abnormally elevated concentration of ambient carbon dioxide (CO2) may
occur under certain environmental conditions in the laboratory. The
Reaction mode R-S fluctuating ambient CO2 concentration may interfere with the CO2L assay
Reaction direction Decrease leading to higher CO2 results. Under these circumstances, the reduction of
the re-calibration interval may become necessary if the laboratory is unable
Wavelength A/B 409/512 nm to keep the ambient CO2 concentration at a normal level by appropriate
Calc. first/last 24/67 countermeasures.
For diagnostic purposes, the results should always be assessed in
Postdilution factor No conjunction with the patients medical history, clinical examination and other
Unit mmol/L findings.
ACTION REQUIRED
Pipetting parameters Special Wash Programming: The use of special wash steps is mandatory
Diluent (H2O) when certain test combinations are run together on COBASINTEGRA
analyzers. Refer to the CLEAN Method Sheet for further instructions and for
R 50 L 120 L the latest version of the Extra wash cycle list.
Sample 2 L 10 L Where required, special wash/carry-over evasion programming must
be implemented prior to reporting results with this test.
Total volume 182 L
Limits and ranges
Calibration Measuring range
2.050mmol/L (2.050mEq/L)
Calibrator Roche Ammonia/Ethanol/CO2 Calibrator
Lower limits of measurement
Use deionized water as zero calibrator. Lower detection limit of the test:
Calibration mode Linear regression 2.0mmol/L (2.0mEq/L)
Calibration replicate Duplicate recommended The lower detection limit represents the lowest measurable analyte level
that can be distinguished from zero. It is calculated as the value lying
Calibration interval Each lot and as required following 3standard deviations above that of the lowest standard (lowest
quality control procedures standard+3SD, repeatability, n=21).
Traceability: This method has been standardized against a primary Expected values
standard. 22-29 mmol/L (22-29 mEq/L)1
Quality control Each laboratory should investigate the transferability of the expected values
to its own patient population and if necessary determine its own reference
Reference range Roche Ammonia/Ethanol/CO2 Control ranges.
Normal or Precinorm U plus Specific performance data
Pathological range Roche Ammonia/Ethanol/CO2 Control Representative performance data on the COBASINTEGRA analyzers are
Abnormal or Precipath U plus given below. Results obtained in individual laboratories may differ.
Control interval 24hours recommended Precision
Precision was determined using human samples and controls in an internal
Control sequence User defined protocol with repeatability (n=21) and intermediate precision (3aliquots
Control after calibration Recommended per run, 1run per day, 21days). The following results were obtained:
For quality control, use control materials as listed in the Order information Level 1 Level 2
section. In addition, other suitable control material can be used.
Mean 19.7 mmol/L 35.4 mmol/L
The control intervals and limits should be adapted to each laboratorys
individual requirements. Values obtained should fall within the defined (19.7 mEq/L) (35.4 mEq/L)
limits. Each laboratory should establish corrective measures to be taken if CV repeatability 0.6 % 0.5 %
values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for Level 1 Level 2
quality control.
Mean 16.8 mmol/L 28.8 mmol/L
Calculation (16.8 mEq/L) (28.8 mEq/L)
COBASINTEGRAanalyzers automatically calculate the analyte
concentration of each sample. For more details, please refer to Data CV intermediate precision 3.5 % 3.8 %
Analysis in the Online Help (COBASINTEGRA400plus/800 analyzers).

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Bicarbonate liquid Substrates

Method comparison 15 Bakker AJ, Mcke M. Gammopathy interference in clinical chemistry

Bicarbonate values for human serum and plasma samples obtained on a assays: mechanisms, detection and prevention.
COBASINTEGRA800 analyzer using the COBASINTEGRA Bicarbonate ClinChemLabMed2007;45(9):1240-1243.
liquid reagent(y) were compared to those determined using the 16 Bablok W, Passing H, Bender R, et al. A general regression procedure
COBASINTEGRA Carbon Dioxide reagent (CO2S) on a for method transformation. Application of linear regression procedures
COBASINTEGRA800 analyzer(x) and using the Bicarbonate liquid assay for method comparison studies in clinical chemistry, Part III. J Clin
on a Roche/Hitachi917 analyzer(x). Chem Clin Biochem 1988 Nov;26(11):783-790.
COBASINTEGRA800 analyzer Sample size (n) = 57 A point (period/stop) is always used in this Method Sheet as the decimal
separator to mark the border between the integral and the fractional parts of
Passing/Bablok16 Linear regression a decimal numeral. Separators for thousands are not used.
y = 0.981x + 0.176 mmol/L y = 0.974x + 0.348 mmol/L Symbols
= 0.984 r = 0.999 Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 152231 standard.
SD (md 95) = 0.400 Sy.x = 0.195
The sample concentrations were between 1.13 and 46.2mmol/L (1.13 and Contents of kit
46.2mEq/L) Volume after reconstitution or mixing
Roche/Hitachi 917 analyzer Sample size (n) = 57 GTIN Global Trade Item Number
Passing/Bablok16 Linear regression
FOR US CUSTOMERS ONLY: LIMITED WARRANTY
y = 1.010x + 0.128 mmol/L y = 1.006x + 0.427 mmol/L Roche Diagnostics warrants that this product will meet the specifications
= 0.969 r = 0.998 stated in the labeling when used in accordance with such labeling and will
SD (md 95) = 1.06 Sy.x = 0.434 be free from defects in material and workmanship until the expiration date
printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
The sample concentrations were between 1.1 and 44.3mmol/L (1.1 and
44.3mEq/L). OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
References PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE
1 Scott MG, Heusel JW, LeGrys VA, et al. Electrolytes and blood gases, FOR INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL
in Tietz NW. Textbook of Clinical Chemistry. 3rd ed. Philadelphia, PA:
WB Saunders Co 1999;1065-1066. DAMAGES.
COBAS, COBASC, COBASINTEGRA, PRECINORM and PRECIPATH are trademarks of Roche.
2 Natelson S. Microtechniques of Clinical Chemistry. Springfield, Il:
All other product names and trademarks are the property of their respective owners.
Charles C Thomas 1975;147.
Significant additions or changes are indicated by a change bar in the margin.
3 Segal MA. A rapid electrotitrimetric method for determining CO2 2014, Roche Diagnostics
combining power in plasma or serum. Am J Clin Pathol
1955;25:1212-1216.
4 Wilson W, Jesyk P, Rand R, et al. Use of Vickers discrete analyzer for
enzymatic determination of the bicarbonate content of serum. Clin Roche Diagnostics GmbH, SandhoferStrasse116, D-68305 Mannheim
Chem 1973;19(6):640. [Link]

5 Norris KA, Atkinson AR, Smith WG, et al. Colorimetric enzymatic Distribution in USA by:
Roche Diagnostics, Indianapolis, IN
determination of serum total carbon dioxide, as applied to the Vickers US Customer Technical Support 1-800-428-2336
Multichannel 300 discrete analyzer. Clin Chem 1975;21:1093-1101.
6 Gambino SR, Schreiber H. The measurement of CO2 content with the
autoanalyzer. A comparison with 3 standard methods and a description
of a new method (alkalinization) for preventing loss of CO2 from open
cups. Am J Clin Path 1966;45:406.
7 Guder WG, da Fonseca-Wollheim F, Heil W, et al. Quality of Diagnostic
Samples, in brochure: Recommendations of the Working Group on
Preanalytical Quality of the German Society for Clinical Chemistry and
Laboratory Medicine, 3rd Ed. 2010.
8 Boyanton BL, Blick KE. Stability studies of Twenty-Four Analytes in
Human Plasma and Serum. Clin Chem 2002;48/12:2242-2247.
9 O'Keane MP, Cunningham SK. Evaluation of three different specimen
types (serum, plasma lithium heparin and serum gel separator) for
analysis of certain analytes: clinical significance of differences in results
and efficiency in use. Clin Chem Lab Med 2006;44(5):662-668.
10 Elfath D, Cooney J, McDaniel R, et al. Effect of frozen storage of serum
on the level of 22 chemistry analytes. Clin Chem 1991;37:931.
11 Tietz NW, ed. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia
PA: WB Saunders Company 1995;84.
12 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation.
ClinChem1986;32:470-475.
13 Breuer J. Report on the Symposium Drug effects in Clinical Chemistry
Methods. Eur J Clin Chem Clin Biochem 1996;34:385-386.
14 Sonntag O, Scholer A. Drug interference in clinical chemistry:
recommendation of drugs and their concentrations to be used in drug
interference studies. Ann Clin Biochem 2001;38:376-385.

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