0003289923190COINV9.

CO2-L
Bicarbonate liquid Substrates
Order information
Analyzer(s) on which cobas c pack(s) can be used
03289923 190 Bicarbonate liquid (250 tests) System-ID 07 6725 5 COBAS INTEGRA 400 plus
20751995 190 Ammonia/Ethanol/CO2 Calibrator (2 × 4 mL) System-ID 07 5199 5
20752401 190 Ammonia/Ethanol/CO2 Control Normal (5 × 4 mL) System-ID 07 5240 1
20753009 190 Ammonia/Ethanol/CO2 Control Abnormal (5 × 4 mL) System-ID 07 5300 9
12149435 122 Precinorm U plus (10 × 3 mL) System-ID 07 7999 7
12149435 160 Precinorm U plus (10 × 3 mL, for USA) System-ID 07 7999 7
12149443 122 Precipath U plus (10 × 3 mL) System-ID 07 8000 6
12149443 160 Precipath U plus (10 × 3 mL, for USA) System-ID 07 8000 6

English Reagent handling

System information Ready for use
Test CO2‑L, test ID 0‑625 Storage and stability
Intended use Shelf life at 2‑8 °C See expiration date on cobas c pack label
In vitro test for the quantitative determination of the bicarbonate (HCO3-)
concentration in human serum and plasma on COBAS INTEGRA systems. On-board in use at 10‑15 °C 6 weeks
Summary Specimen collection and preparation
Bicarbonate is the second largest fraction of the anions in plasma. Included For specimen collection and preparation only use suitable tubes or
in this fraction are the bicarbonate (HCO3-) and carbonate (CO32-) ions, as collection containers.
well as the carbamino compounds. At the physiological pH of blood, the Only the specimens listed below were tested and found acceptable.
concentration of carbonate is 1/1000 that of bicarbonate. The carbamino Serum
compounds are also present in such low quantities that they are generally Plasma: Heparin (Li-, Na-, NH4+-) plasma
not mentioned specifically. The preferred specimen is from venous blood collected anaerobically in the
Several different methods for the determination of bicarbonate in serum and usual manner for bicarbonate analysis. Bicarbonate content in uncapped
plasma have been reported. Most of these procedures utilize acidification of tubes decreases approximately 4 mmol/L after one hour.6 It has been
the sample and conversion of all carbon dioxide forms to CO2 gas.1 The reported that alkalinized serum stored in open cups is stable for up to
amount of gas formed is measured by manometric or volumetric devices, 4 hours.6
ion selective electrodes, or spectrophotometric techniques.2,3 These The sample types listed were tested with a selection of sample collection
methods are either cumbersome, time‑consuming, technique‑oriented, tubes that were commercially available at the time of testing, i.e. not all
and/or require special equipment. available tubes of all manufacturers were tested. Sample collection systems
Enzymatic procedures using phosphoenolpyruvate carboxylase (PEPC) from various manufacturers may contain differing materials which could
have been described.4,5 affect the test results in some cases. When processing samples in primary
The bicarbonate content of serum or plasma is a significant indicator of tubes (sample collection systems), follow the instructions of the tube
electrolyte dispersion and anion deficit. Together with pH determination, manufacturer.
bicarbonate measurements are used in the diagnosis and treatment of Centrifuge samples containing precipitates before performing the assay.
numerous potentially serious disorders associated with acid‑base Separate from erythrocytes and store tightly stoppered.
imbalance in the respiratory and metabolic systems.
See the limitations and interferences section for details about possible
Test principle sample interferences.
Bicarbonate reacts with phosphoenolpyruvate (PEP) in the presence of
PEPC to produce oxaloacetate and phosphate: Stability: 7 days at 4‑8 °C7
40 hours at 15‑25 °C8,9
PEPC

PEP + HCO3- oxaloacetate + H2PO4- Storage of serum at ‑20 °C or ‑80 °C for up to 6 months had no significant
effect.10
The above reaction is coupled with one involving the transfer of a hydrogen Sample stability claims were established by experimental data by the
ion from NADH analog to oxaloacetate using MDH. manufacturer or based on reference literature and only for the
temperatures/time frames as stated in the method sheet. It is the
MDH responsibility of the individual laboratory to use all available references
Oxaloacetate + NADH analog + H+ malate + NAD+ analog and/or its own studies to determine specific stability criteria for its
laboratory.
The resultant consumption of NADH analog causes a decrease in
absorbance at 409 nm, which is proportional to the concentration of Materials provided
bicarbonate in the sample being assayed. See “Reagents – working solutions” section for reagents.
Reagents - working solutions Assay
For optimum performance of the assay follow the directions given in this
R Phosphoenolpyruvate: ≥ 40 mmol/L; NADH analog: ≥ 2 mmol/L; document for the analyzer concerned. Refer to the appropriate operator’s
MDH (porcine): ≥ 314.3 µkat/L; PEPC (microbial): ≥ 30.8 µkat/L manual for analyzer‑specific assay instructions.
R is in position B. Application for serum and plasma
Precautions and warnings Test definition
Pay attention to all precautions and warnings listed in
Section 1 / Introduction of this Method Manual. Measuring mode Absorbance
For USA: Caution: Federal law restricts this device to sale by or on the Abs. calculation mode Endpoint
order of a physician.
Reaction mode R-S

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Bicarbonate liquid Substrates

Reaction direction Decrease Immunoglobulins: No significant interference from immunoglobulins up to a

concentration of 35 g/L (233.5 µmol/L) (simulated by human
Wavelength A/B 409/512 nm immunoglobulin G)
Calc. first/last 21/45 In very rare cases, gammopathy, in particular type IgM (Waldenström’s
Unit mmol/L macroglobulinemia), may cause unreliable results.15
An abnormally elevated concentration of ambient carbon dioxide (CO2) may
Pipetting parameters occur under certain environmental conditions in the laboratory. The
fluctuating ambient CO2 concentration may interfere with the CO2‑L assay
Diluent (H2O) leading to higher CO2 results. Under these circumstances, the reduction of
R 50 µL 120 µL the re-calibration interval may become necessary if the laboratory is unable
to keep the ambient CO2 concentration at a normal level by appropriate
Sample 2 µL 10 µL countermeasures.
Total volume 182 µL For diagnostic purposes, the results should always be assessed in
conjunction with the patient’s medical history, clinical examination and other
Calibration findings.
Calibrator Roche Ammonia/Ethanol/CO2 Calibrator ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory
Use deionized water as zero calibrator. when certain test combinations are run together on COBAS INTEGRA
Calibration mode Linear regression analyzers. Refer to the CLEAN Method Sheet for further instructions and for
the latest version of the Extra wash cycle list.
Calibration replicate Duplicate recommended Where required, special wash/carry-over evasion programming must
be implemented prior to reporting results with this test.
Calibration interval Each lot and as required following
quality control procedures Limits and ranges
Calibration interval may be extended based on acceptable verification of Measuring range
calibration by the laboratory. 2.0‑50 mmol/L (2.0‑50 mEq/L)
Traceability: This method has been standardized against a primary Lower limits of measurement
standard. Lower detection limit of the test:
2.0 mmol/L (2.0 mEq/L)
Quality control
The lower detection limit represents the lowest measurable analyte level
Reference range Roche Ammonia/Ethanol/CO2 Control that can be distinguished from zero. It is calculated as the value lying
Normal or Precinorm U plus 3 standard deviations above that of the lowest standard (lowest
standard + 3 SD, repeatability, n = 21).
Pathological range Roche Ammonia/Ethanol/CO2 Control
Abnormal or Precipath U plus Expected values
22-29 mmol/L (22-29 mEq/L)1
Control interval 24 hours recommended
Each laboratory should investigate the transferability of the expected values
Control sequence User defined to its own patient population and if necessary determine its own reference
ranges.
Control after calibration Recommended
Specific performance data
For quality control, use control materials as listed in the “Order information”
section. In addition, other suitable control material can be used. Representative performance data on the COBAS INTEGRA analyzers are
given below. Results obtained in individual laboratories may differ.
The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined Precision
limits. Each laboratory should establish corrective measures to be taken if Precision was determined using human samples and controls in an internal
values fall outside the defined limits. protocol with repeatability (n = 21) and intermediate precision (3 aliquots
Follow the applicable government regulations and local guidelines for per run, 1 run per day, 21 days). The following results were obtained:
quality control.
Level 1 Level 2
Calculation
Mean 19.7 mmol/L 35.4 mmol/L
The COBAS INTEGRA 400 plus analyzer automatically calculates the
analyte concentration of each sample. For more details, please refer to (19.7 mEq/L) (35.4 mEq/L)
Data Analysis in the Online Help. CV repeatability 0.6 % 0.5 %
Conversion factor: mmol/L × 1 = mEq/L11
Limitations - interference Level 1 Level 2
Criterion: Recovery within ± 10 % of initial value. Mean 16.8 mmol/L 28.8 mmol/L
Icterus:12 No significant interference up to an I index of 60 for conjugated (16.8 mEq/L) (28.8 mEq/L)
and unconjugated bilirubin (approximate conjugated and unconjugated CV intermediate precision 3.5 % 3.8 %
bilirubin concentration: 1026 µmol/L or 60 mg/dL).
Hemolysis:12 No significant interference up to an H index of 1000 Method comparison
(approximate hemoglobin concentration: 621 µmol/L or 1000 mg/dL). Bicarbonate values for human serum and plasma samples obtained on a
Lipemia (Intralipid):12 No significant interference up to an L index of 2000. COBAS INTEGRA 800 analyzer using the COBAS INTEGRA Bicarbonate
There is poor correlation between the L index (corresponds to turbidity) and liquid reagent (y) were compared with those determined using the
triglycerides concentration. COBAS INTEGRA Carbon Dioxide reagent (CO2‑S) on a
COBAS INTEGRA 800 analyzer (x) and using the Bicarbonate liquid assay
Drugs: No interference was found at therapeutic concentrations using on a Roche/Hitachi 917 analyzer (x).
common drug panels.13,14
Criterion: Recovery within ± 10 % of initial value at a bicarbonate COBAS INTEGRA 800 analyzer Sample size (n) = 57
concentration of 22 mmol/L.
Passing/Bablok16 Linear regression
y = 0.981x + 0.176 mmol/L y = 0.973x + 0.355 mmol/L

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Bicarbonate liquid Substrates

τ = 0.984 r = 1.000 Symbols

SD (md 95) = 0.400 Sy.x = 0.195 Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard (for USA: see dialog.roche.com for
The sample concentrations were between 1.13 and 46.2 mmol/L (1.13 and definition of symbols used):
46.2 mEq/L)
Contents of kit
Roche/Hitachi 917 analyzer Sample size (n) = 57
Volume after reconstitution or mixing
Passing/Bablok16 Linear regression
GTIN Global Trade Item Number
y = 1.010x + 0.128 mmol/L y = 1.004x + 0.467 mmol/L
τ = 0.969 r = 0.998 FOR US CUSTOMERS ONLY: LIMITED WARRANTY
SD (md 95) = 1.06 Sy.x = 0.434 Roche Diagnostics warrants that this product will meet the specifications
stated in the labeling when used in accordance with such labeling and will
The sample concentrations were between 1.1 and 44.3 mmol/L (1.1 and be free from defects in material and workmanship until the expiration date
44.3 mEq/L).
printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
References OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
1 Scott MG, Heusel JW, LeGrys VA, et al. Electrolytes and blood gases, WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
in Tietz NW. Textbook of Clinical Chemistry. 3rd ed. Philadelphia, PA:
WB Saunders Co 1999;1065-1066. PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE
FOR INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL
2 Natelson S. Microtechniques of Clinical Chemistry. Springfield, Il: DAMAGES.
Charles C Thomas 1975;147.
COBAS, COBAS C, COBAS INTEGRA, PRECINORM and PRECIPATH are trademarks of Roche.
3 Segal MA. A rapid electrotitrimetric method for determining CO2 All other product names and trademarks are the property of their respective owners.
combining power in plasma or serum. Am J Clin Pathol Additions, deletions or changes are indicated by a change bar in the margin.
1955;25:1212-1216. © 2020, Roche Diagnostics
4 Wilson W, Jesyk P, Rand R, et al. Use of Vickers discrete analyzer for
enzymatic determination of the bicarbonate content of serum. Clin
Chem 1973;19(6):640.
5 Norris KA, Atkinson AR, Smith WG, et al. Colorimetric enzymatic Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
determination of serum total carbon dioxide, as applied to the Vickers www.roche.com
Multichannel 300 discrete analyzer. Clin Chem 1975;21:1093-1101. Distribution in USA by:
Roche Diagnostics, Indianapolis, IN
6 Gambino SR, Schreiber H. The measurement of CO2 content with the US Customer Technical Support 1-800-428-2336
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.
Clin Chem 1986;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.
15 Bakker AJ, Mücke M. Gammopathy interference in clinical chemistry
assays: mechanisms, detection and prevention.
Clin Chem Lab Med 2007;45(9):1240-1243.
16 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
for method comparison studies in clinical chemistry, Part III. J Clin
Chem Clin Biochem 1988 Nov;26(11):783-790.
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
a decimal numeral. Separators for thousands are not used.

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