0005061482190c501V5.

CA2
Calcium Gen.2
Order information
Analyzer(s) on which cobas c pack(s) can be used
05061482 190 Calcium Gen.2 (300 tests) System‑ID 07 7476 6 Roche/Hitachi cobas c 311, cobas c 501/502
10759350 190 Calibrator f.a.s. (12 × 3 mL) Code 401
10759350 360 Calibrator f.a.s. (12 × 3 mL, for USA) Code 401
12149435 122 Precinorm U plus (10 × 3 mL) Code 300
12149435 160 Precinorm U plus (10 × 3 mL, for USA) Code 300
12149443 122 Precipath U plus (10 × 3 mL) Code 301
12149443 160 Precipath U plus (10 × 3 mL, for USA) Code 301
05117003 190 PreciControl ClinChem Multi 1 (20 × 5 mL) Code 391
05947626 190 PreciControl ClinChem Multi 1 (4 × 5 mL) Code 391
05947626 160 PreciControl ClinChem Multi 1 (4 × 5 mL, for USA) Code 391
05117216 190 PreciControl ClinChem Multi 2 (20 × 5 mL) Code 392
05947774 190 PreciControl ClinChem Multi 2 (4 × 5 mL) Code 392
05947774 160 PreciControl ClinChem Multi 2 (4 × 5 mL, for USA) Code 392
04489357 190 Diluent NaCl 9 % (50 mL) System‑ID 07 6869 3

English a) 3‑[cyclohexylamino]‑2‑hydroxy‑1‑propanesulfonic acid

System information R1 is in position B and R2 is in position C.

For cobas c 311/501 analyzers: Precautions and warnings
CA2: ACN 698 For in vitro diagnostic use.
S‑CA2: ACN 699 (STAT, reaction time: 3) Exercise the normal precautions required for handling all laboratory
reagents.
For cobas c 502 analyzer: Disposal of all waste material should be in accordance with local guidelines.
CA2: ACN 8698 Safety data sheet available for professional user on request.
S‑CA2: ACN 8699 (STAT, reaction time: 3) For USA: Caution: Federal law restricts this device to sale by or on the
order of a physician.
Intended use
In vitro test for the quantitative determination of calcium in human serum, Reagent handling
plasma and urine on Roche/Hitachi cobas c systems. Ready for use
Summary1 Storage and stability
Calcium is the most abundant mineral element in the body with about
99 percent in the bones primarily as hydroxyapatite. The remaining calcium CA2
is distributed between the various tissues and the extracellular fluids where Shelf life at 2‑8 °C: See expiration date
it performs a vital role for many life sustaining processes. Among the extra on cobas c pack
skeletal functions of calcium are involvement in blood coagulation,
neuromuscular conduction, excitability of skeletal and cardiac muscle, label.
enzyme activation, and the preservation of cell membrane integrity and On‑board in use and refrigerated on the analyzer: 6 weeks
permeability.
Diluent NaCl 9 %
Serum calcium levels and hence the body content are controlled by
parathyroid hormone (PTH), calcitonin, and vitamin D. An imbalance in any Shelf life at 2‑8 °C: See expiration date
of these modulators leads to alterations of the body and serum calcium on cobas c pack
levels. Increases in serum PTH or vitamin D are usually associated with label.
hypercalcemia. Increased serum calcium levels may also be observed in
multiple myeloma and other neoplastic diseases. Hypocalcemia may be On‑board in use and refrigerated on the analyzer: 12 weeks
observed e.g. in hypoparathyroidism, nephrosis, and pancreatitis.
Specimen collection and preparation
Test principle For specimen collection and preparation only use suitable tubes or
Calcium ions react with 5‑nitro‑5’‑methyl‑BAPTA (NM-BAPTA) under collection containers.
alkaline conditions to form a complex. This complex reacts in the second Only the specimens listed below were tested and found acceptable.
step with EDTA. Serum: Fresh serum collected in the fasting state is the preferred specimen.
Plasma: Li‑heparin plasma.
alkaline pH
Serum or plasma should be separated from blood cells as soon as possible,
Ca2+ + NM‑BAPTA calcium‑NM‑BAPTA complex because prolonged contact with the clot may cause lower calcium values.2
Sera from patients receiving EDTA (treatment of hypercalcemia) are
calcium‑NM‑BAPTA complex NM‑BAPTA unsuitable for analysis, since EDTA will chelate the calcium and render it
+ EDTA + calcium EDTA complex unavailable for reaction with NM‑BAPTA. Co-precipitation of calcium with
fibrin (i.e. heparin plasma), lipids, or denatured protein has been reported
The change in absorbance is directly proportional to the calcium with storage or freezing.1,3
concentration and is measured photometrically.
The sample types listed were tested with a selection of sample collection
Reagents - working solutions tubes that were commercially available at the time of testing, i.e. not all
available tubes of all manufacturers were tested. Sample collection systems
R1 CAPSO:a 557 mmol/L; NM‑BAPTA: 2 mmol/L; pH 10.0; from various manufacturers may contain differing materials which could
non‑reactive surfactant; preservative affect the test results in some cases. When processing samples in primary
R2 EDTA: 7.5 mmol/L; pH 7.3; non‑reactive surfactant, preservative tubes (sample collection systems), follow the instructions of the tube
manufacturer.

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Calcium Gen.2

Urine Reagent pipetting Diluent (H2O)

Urine specimens should be collected in acid‑washed bottles. 24‑hour R1 20 µL 160 µL
specimens should be collected in containers containing 20‑30 mL of 6 mol/L
HCl to prevent calcium salt precipitation. Precipitated calcium salts may not R2 20 µL - 
be completely dissolved by the addition of HCl following urine collection.4
Stability in serum/plasma:5 7 days at 15‑25 °C Sample volumes Sample Sample dilution
3 weeks at 2‑8 °C Sample Diluent (NaCl)
8 months at (-15)‑(-25) °C Normal 3 µL – –
Decreased 3 µL – –
Stability in urine:5 2 days at 15‑25 °C
Increased 3 µL – –
4 days at 2‑8 °C
Application for urine
3 weeks at (-15)‑(-25) °C
Stored serum or urine specimens must be mixed well prior to analysis. cobas c 311 test definition
Centrifuge samples containing precipitates before performing the assay. Assay type 2‑Point End
See the limitations and interferences section for details about possible Reaction time / Assay points 10 / 6‑8 (STAT 3 / 6‑8)
sample interferences.
Wavelength (sub/main) 376/340 nm
Sample stability claims were established by experimental data by the
manufacturer or based on reference literature and only for the Reaction direction Decrease
temperatures/time frames as stated in the method sheet. It is the
responsibility of the individual laboratory to use all available references Units mmol/L (mg/dL)
and/or its own studies to determine specific stability criteria for its Reagent pipetting Diluent (H2O)
laboratory.
R1 20 µL 160 µL
Materials provided
R2 20 µL - 
See “Reagents – working solutions” section for reagents.
Materials required (but not provided)
▪ See “Order information” section Sample volumes Sample Sample dilution
▪ General laboratory equipment Sample Diluent (NaCl)
Assay Normal 2 µL – –
For optimum performance of the assay follow the directions given in this Decreased 4 µL 15 µL 135 µL
document for the analyzer concerned. Refer to the appropriate operator’s
manual for analyzer‑specific assay instructions. Increased 2 µL – –
The performance of applications not validated by Roche is not warranted cobas c 501/502 test definition
and must be defined by the user.
Assay type 2‑Point End
Application for serum and plasma
Reaction time / Assay points 10 / 10‑13 (STAT 3 / 10‑13)
cobas c 311 test definition
Wavelength (sub/main) 376/340 nm
Assay type 2‑Point End
Reaction direction Decrease
Reaction time / Assay points 10 / 6‑8 (STAT 3 / 6‑8)
Units mmol/L (mg/dL)
Wavelength (sub/main) 376/340 nm
Reaction direction Decrease
Reagent pipetting Diluent (H2O)
Units mmol/L (mg/dL)
R1 20 µL 160 µL
Reagent pipetting Diluent (H2O)
R2 20 µL - 
R1 20 µL 160 µL
R2 20 µL - 
Sample volumes Sample Sample dilution
Sample Diluent (NaCl)
Sample volumes Sample Sample dilution
Normal 2 µL – –
Sample Diluent (NaCl)
Decreased 4 µL 15  135 
Normal 3 µL – –
Increased 2 µL – –
Decreased 3 µL – –
Calibration
Increased 3 µL – –
Calibrators S1: H2O
cobas c 501/502 test definition
S2: C.f.a.s.
Assay type 2‑Point End
Calibration mode Linear
Reaction time / Assay points 10 / 10‑13 (STAT 3 / 10‑13)
Calibration frequency 2‑point calibration
Wavelength (sub/main) 376/340 nm • after reagent lot change
Reaction direction Decrease • as required following quality control
Units mmol/L (mg/dL) procedures

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Calcium Gen.2

Calibration interval may be extended based on acceptable verification of For diagnostic purposes, the results should always be assessed in
calibration by the laboratory. conjunction with the patient’s medical history, clinical examination and other
Traceability: This method has been standardized against the findings.
SRM 956 c Level 2 reference material. ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory
Quality control when certain test combinations are run together on Roche/Hitachi
Serum/plasma cobas c systems. The latest version of the carry‑over evasion list can be
For quality control, use control materials as listed in the "Order information" found with the NaOHD-SMS-SmpCln1+2-SCCS Method Sheets. For further
section. instructions refer to the operator’s manual. cobas c 502 analyzer: All
In addition, other suitable control material can be used. special wash programming necessary for avoiding carry‑over is available
via the cobas link, manual input is required in certain cases.
Urine
Where required, special wash/carry‑over evasion programming must
Quantitative urine controls are recommended for routine quality control. be implemented prior to reporting results with this test.
The control intervals and limits should be adapted to each laboratory’s Limits and ranges
individual requirements. Values obtained should fall within the defined
limits. Each laboratory should establish corrective measures to be taken if Measuring range
values fall outside the defined limits. Serum/plasma
Follow the applicable government regulations and local guidelines for 0.20‑5.0 mmol/L (0.8‑20.1 mg/dL)
quality control. Urine
Calculation 0.20‑7.5 mmol/L (0.8‑30.1 mg/dL)
Roche/Hitachi cobas c systems automatically calculate the analyte Determine urine samples having higher concentrations via the rerun
concentration of each sample. function. Dilution of samples via the rerun function is a 1:5 dilution. Results
from samples diluted using the rerun function are automatically multiplied
Conversion factors: mmol/L × 4.01 = mg/dL by a factor of 5.
In studies with 24‑hour urine, multiply the value obtained by the 24‑hour Lower limits of measurement
volume in order to obtain a measurement in mg/24 h or mmol/24 h. Limit of Blank, Limit of Detection and Limit of Quantitation
Limitations - interference Serum/plasma and urine
Criterion: Recovery within ± 0.22 mmol/L (0.9 mg/dL) of initial value of
samples ≤ 2.2 mmol/L (8.8 mg/dL) and within ± 10 % for samples Serum/plasma
> 2.2 mmol/L. Limit of Blank: = 0.10 mmol/L (0.4 mg/dL)
Serum/plasma
Limit of Detection: = 0.20 mmol/L (0.8 mg/dL)
Icterus:6 No significant interference up to an I index of 60 for conjugated
and unconjugated bilirubin (approximate conjugated and unconjugated Limit of Quantitation = 0.20 mmol/L (0.8 mg/dL)
bilirubin concentration: 1026 µmol/L or 60 mg/dL). The Limit of Blank, Limit of Detection and Limit of Quantitation were
Hemolysis:6 No significant interference up to an H index of 1000 determined in accordance with the CLSI (Clinical and Laboratory Standards
(approximate hemoglobin concentration: 621 µmol/L or 1000 mg/dL). Institute) EP17‑A2 requirements.
Lipemia (Intralipid):6 No significant interference up to an L index of 1000. The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of
There is a poor correlation between the L index (corresponds to turbidity) analyte‑free samples over several independent series. The Limit of Blank
and triglycerides concentration. corresponds to the concentration below which analyte‑free samples are
Magnesium: No significant interference from magnesium up to a found with a probability of 95 %.
concentration of 15 mmol/L (36.5 mg/dL). The Limit of Detection is determined based on the Limit of Blank and the
Drugs: No interference was found at therapeutic concentrations using standard deviation of low concentration samples.
common drug panels.7,8 The Limit of Detection corresponds to the lowest analyte concentration
The interference of intravenously administered gadolinium containing MRI which can be detected (value above the Limit of Blank with a probability of
(magnetic resonance imaging) contrast media was tested (Omniscan®, 95 %).
Optimark®) but no interference was found at the therapeutic concentration. The Limit of Quantitation is the lowest analyte concentration that can be
Interferences at higher concentrations were observed. reproducibly measured with a total error of 30 %. It has been determined
In very rare cases, gammopathy, in particular type IgM (Waldenström’s using low concentration calcium samples.
macroglobulinemia), may cause unreliable results.9 Expected values10
Urine
Serum/plasma
Icterus: No significant interference up to a conjugated bilirubin
concentration of 1026 μmol/L or 60 mg/dL. Children (0‑10 days): 1.90‑2.60 mmol/L (7.6‑10.4 mg/dL)
Hemolysis: No significant interference up to a hemoglobin concentration of Children (10 days‑2 years): 2.25‑2.75 mmol/L (9.0‑11.0 mg/dL)
621 μmol/L or 1000 mg/dL.
Children (2‑12 years): 2.20‑2.70 mmol/L (8.8‑10.8 mg/dL)
Magnesium: No significant interference from magnesium up to a
concentration of 60 mmol/L (145.8 mg/dL). Children (12‑18 years): 2.10‑2.55 mmol/L (8.4‑10.2 mg/dL)
Urea: No significant interference from urea up to a concentration of Adults (18‑60 years): 2.15‑2.50 mmol/L (8.6‑10.0 mg/dL)
1600 mmol/L (9610 mg/dL).
Adults (60‑90 years): 2.20‑2.55 mmol/L (8.8‑10.2 mg/dL)
Drugs: No interference was found at therapeutic concentrations using
common drug panels.8 Adults ( > 90 years): 2.05‑2.40 mmol/L (8.2‑9.6 mg/dL)
The interference of intravenously administered gadolinium containing MRI Urine
(magnetic resonance imaging) contrast media was tested (Omniscan®, 2.5‑7.5 mmol/24 h (100‑300 mg/24 h) with normal food intake.
Optimark®). For Omniscan® no interference was observed at the therapeutic
concentration, but there was interference at higher concentrations. For Roche has not evaluated reference ranges in a pediatric population.
Optimark® interference was observed at therapeutic and higher Each laboratory should investigate the transferability of the expected values
concentrations. to its own patient population and if necessary determine its own reference
ranges.

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Calcium Gen.2

Specific performance data The sample concentrations were between 0.33 and 4.76 mmol/L (1.3 and
Representative performance data on the analyzers are given below. 19.1 mg/dL).
Results obtained in individual laboratories may differ.
Precision Urine
Repeatability and intermediate precision were determined using human Sample size (n) = 65
samples and controls in accordance with the CLSI (Clinical and Laboratory Passing/Bablok11 Linear regression
Standards Institute) EP5 requirements (2 aliquots per run, 2 runs per day,
21 days). The following results were obtained: y = 0.989x + 0.064 mmol/L y = 0.983x + 0.079 mmol/L
Serum/plasma τ = 0.989 r = 1.00
Repeatability Mean SD CV The sample concentrations were between 0.28 and 7.47 mmol/L (1.1 and
30.0 mg/dL).
mmol/L (mg/dL) mmol/L (mg/dL) %
Calcium values for human serum, plasma and urine samples obtained on a
Human serum 1 0.60 (2.4) 0.01 (0.0) 2.0 Roche/Hitachi cobas c 501 analyzer using the Roche Calcium Gen.2
Human serum 2 2.55 (10.2) 0.02 (0.1) 0.8 reagent (y) were compared with those determined using the Roche Calcium
reagent on a Roche/Hitachi MODULAR P analyzer (x).
Human serum 3 4.46 (17.9) 0.04 (0.2) 0.8
Precinorm U 2.25 (9.0) 0.02 (0.1) 0.8 Serum/plasma
Precipath U 3.51 (14.1) 0.03 (0.1) 0.8 Sample size (n) = 69
Passing/Bablok11 Linear regression
Intermediate Mean SD CV
y = 1.018x - 0.027 mmol/L y = 1.023x - 0.036 mmol/L
precision mmol/L (mg/dL) mmol/L (mg/dL) %
τ = 0.976 r = 1.00
Human serum 1 0.60 (2.4) 0.02 (0.1) 2.5
The sample concentrations were between 0.28 and 4.65 mmol/L (1.1 and
Human serum 2 2.55 (10.2) 0.02 (0.1) 0.9 18.6 mg/dL).
Human serum 3 4.46 (17.9) 0.04 (0.2) 0.9
Urine
Precinorm U 2.25 (9.0) 0.02 (0.1) 0.8
Sample size (n) = 65
Precipath U 3.51 (14.1) 0.03 (0.1) 0.9
Passing/Bablok11 Linear regression
Urine
y = 1.024x + 0.018 mmol/L y = 1.020x + 0.029 mmol/L
Repeatability Mean SD CV τ = 0.988 r = 1.00
mmol/L (mg/dL) mmol/L (mg/dL) % The sample concentrations were between 0.30 and 7.25 mmol/L (1.2 and
Human urine 1 0.58 (2.3) 0.02 (0.1) 3.0 29.1 mg/dL).
Human urine 2 3.92 (15.7) 0.04 (0.2) 1.1 References
Human urine 3 5.18 (20.8) 0.05 (0.2) 0.9 1 Endres DB, Rude RK. Mineral and Bone Metabolism. In: Burtis CA,
Ashwood ER, Bruns ED, eds. Tietz Textbook of Clinical Chemistry and
Human urine 4 6.09 (24.4) 0.08 (0.3) 1.3 Molecular Diagnostics, 4th ed. St. Louis (MO): Saunders Elsevier
Control Level 1 1.85 (7.4) 0.02 (0.1) 1.3 2006:1891-1965.
Control Level 2 2.72 (10.9) 0.03 (0.1) 1.1 2 Heins M, Heil W, Withold W. Storage of Serum or Whole Blood
Samples? Effect of Time and Temperature on 22 Serum Analytes. Eur
J Clin Chem Clin Biochem 1995;33:231-238.
Intermediate Mean SD CV
precision 3 Wilding P, Zilva JF, Wilde CE. Transport of specimens for clinical
mmol/L (mg/dL) mmol/L (mg/dL) % chemistry analysis. Ann Clin Biochem 1977;14:301-306.
Human urine 1 0.58 (2.3) 0.02 (0.1) 3.1 4 Burtis CA, Ashwood ER, Bruns DE, eds. Tietz Fundamentals of Clinical
Human urine 2 3.92 (15.7) 0.05 (0.2) 1.2 Chemistry, 6th ed.St. Louis (MO): Saunders Elsevier 2008:715.
5 Use of Anticoagulants in Diagnostic Laboratory Investigations. WHO
Human urine 3 5.18 (20.8) 0.06 (0.2) 1.1 Publication WHO/DIL/LAB/99.1 Rev. 2: Jan 2002.
Human urine 4 6.09 (24.4) 0.08 (0.3) 1.3 6 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Control Level 1 1.85 (7.4) 0.03 (0.1) 1.5 Interferences in Clinical Chemistry Instrumentation. Clin Chem
1986;32:470-475.
Control Level 2 2.72 (10.9) 0.04 (0.2) 1.3
7 Breuer J. Report on the Symposium “Drug effects in Clinical Chemistry
Method comparison Methods”. Eur J Clin Chem Clin Biochem 1996;34:385-386.
Calcium values for human serum, plasma and urine samples obtained on a 8 Sonntag O, Scholer A. Drug interference in clinical chemistry:
Roche/Hitachi cobas c 501 analyzer using the Roche Calcium Gen.2 recommendation of drugs and their concentrations to be used in drug
reagent (x) were compared with those determined using the corresponding interference studies. Ann Clin Biochem 2001;38:376-385.
reagent on a Roche/Hitachi MODULAR P analyzer (y).
9 Bakker AJ, Mücke M. Gammopathy interference in clinical chemistry
Serum/plasma assays: mechanisms, detection and prevention.
Clin Chem Lab Med 2007;45(9):1240-1243.
Sample size (n) = 69
10 Wu AHB, ed. Tietz Clinical Guide to Laboratory Tests, 4th ed. St. Louis
Passing/Bablok11 Linear regression (MO): Saunders Elsevier 2006:202-207.
y = 0.982x + 0.061 mmol/L y = 0.982x + 0.059 mmol/L 11 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
τ = 0.979 r = 1.00 for method comparison studies in clinical chemistry, Part III. J Clin
Chem Clin Biochem 1988 Nov;26(11):783-790.

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Calcium Gen.2

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.
Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard (for USA: see
https://usdiagnostics.roche.com for definition of symbols used):
Contents of kit
Volume after reconstitution or mixing
GTIN Global Trade Item Number

FOR US CUSTOMERS ONLY: LIMITED WARRANTY

Roche Diagnostics warrants that this product will meet the specifications
stated in the labeling when used in accordance with such labeling and will
be free from defects in material and workmanship until the expiration date
printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE
FOR INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL
DAMAGES.
COBAS, COBAS C, PRECICONTROL, PRECINORM and PRECIPATH are trademarks of Roche.
All other product names and trademarks are the property of their respective owners.
Additions, deletions or changes are indicated by a change bar in the margin.
© 2018, Roche Diagnostics

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www.roche.com
Distribution in USA by:
Roche Diagnostics, Indianapolis, IN
US Customer Technical Support 1-800-428-2336

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