Siemens Healthcare Diagnostics Inc.

Instructions for Use

2016-06

Calcium_2 (CA_2)
System New Information

ADVIA® 1200 Updates: Expected Values; References

ADVIA 1650/1800 Updates: Expected Values; References

ADVIA 2400 Updates: Expected Values; References

Method Summary
Item Description
Method Principle Arsenazo III
Specimen Type Human serum, plasma (lithium heparin), and urine
On-board Stability ADVIA 1200: 30 days
ADVIA 1650/1800: 30 days
ADVIA 2400: 30 days
Reagent Storage 15–25°C
Temperature
Calibration Frequency ADVIA 1200: 24 days
ADVIA 1650/1800: 24 days
ADVIA 2400: 24 days
Reagent Blank (RBL) At time of method calibration
Frequency
Reaction Type Endpoint (EPA)
Measurement Wavelength 658/694 nm
Standardization Siemens internal reference method (ICP atomic emission)
Analytical Range System Serum/Plasma Urine
ADVIA 1200: 1.0–16.0 mg/dL 1.0–32.0 mg/dL
(0.25–4.00 mmol/L) (0.25–8.00 mmol/L)
ADVIA 1650/1800: 1.0–16.0 mg/dL 1.0–32.0 mg/dL
(0.25–4.00 mmol/L) (0.25–8.00 mmol/L)
ADVIA 2400: 1.0–16.0 mg/dL 1.0–32.0 mg/dL
(0.25–4.00 mmol/L) (0.25–8.00 mmol/L)

© 2009 Siemens Healthcare Diagnostics. All rights reserved.

Siemens Healthcare Diagnostics Inc., Tarrytown, NY 10591-5097 USA
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Item Description
Expected Values Serum/Plasma Urine
8.7–10.4 mg/dL 100–300 mg/day
(2.18–2.60 mmol/L) (2.5–7.5 mmol/day)
Reagent Code 74076
Calibrator Siemens Chemistry Calibrator:
REF 09784096 (T03-1291-62)

Intended Use
For in vitro diagnostic use in the quantitative determination of calcium in human serum,
plasma, and urine on the ADVIA Chemistry systems. Such measurements are used in
the diagnosis and treatment of parathyroid disease, a variety of bone diseases, chronic
renal failure, and tetany.

Summary and Explanation 1

The Calcium_2 (CA_2) method is based on the work of Michaylova and Illkova, who
found that Arsenazo III could form a stable complex with calcium with high selectivity at
low pH.

Principles of the Procedure

Calcium ions form a colored complex with Arsenazo III, which is measured at 658/694
nm. The amount of calcium present in the sample is directly proportional to the intensity
of the colored complex formed.

Reaction Equation

Ca2+ + Arsenazo III Ca-Arsenazo III Complex (purple)

Reagents
The reagents are packaged as listed below. Components of the package are available
as a kit only.

REF
Container Size Symbol Contents Amount No. of Tests
02189915 Calcium 7 x 290
Reagents (ADVIA 1650/1800/2400)
40-mL Reagent 7 x 38 mL 7 x 230 (ADVIA 1200)

02189699 Calcium 8 x 550

Reagents (ADVIA 1650/1800/2400)
70-mL Reagent 8 x 68 mL 8 x 430 (ADVIA 1200)

Safety data sheets (MSDS/SDS) available on siemens.com/healthcare.

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Components and Concentrations

Reagent Component Concentration

Reagent 1 Sodium acetate, pH 5.9 54.2 mmol/L
Arsenazo III 188 µmol/L
Non-reactive stabilizers

For in vitro diagnostic use.

Reagent Preparation and Use

Reagents are ready to use. Before use, gently swirl the reagent to disrupt bubbles and
assure homogeneity. If bubbles still exist or foam is present, using a clean transfer
pipette, aspirate them from the reagent container prior to use.

The ADVIA 1200 Chemistry system automatically predilutes urine samples. Place a
70-mL reagent container (REF 06397121; 073-0373-02) filled with saline (0.9% NaCl)
into RTT 1, position 1. Refill this reagent container once per shift, or as needed.
Remove the container from the system weekly and clean or replace it with a new
container.

On-board Reagent Stability (OBS)

System Stability
ADVIA 1200 30 days
ADVIA 1650/1800 30 days
ADVIA 2400 30 days

For all systems, unopened reagents are stable until the expiration date printed on the
product label when stored at 15° to 25°C. Do not freeze reagents.

Sample Handling
CAUTION! Do not use sodium oxalate, EDTA, or sodium fluoride as anticoagulants, as
they have been found to interfere.

Siemens Healthcare Diagnostics recommends using serum, plasma (lithium heparin),

or urine for this method.

Materials Required but not Provided

The following list contains the materials required, but not provided, to perform this
method:
• sample containers
• system solutions
• calibrator (refer to the Method Summary section for the REFs)
• control materials (refer to the Quality Control section)
• 70-mL reagent container (REF 06397121; 073-0373-02)
• 40-mL adapter (REF 08163594; 073-0788-01) for 70-mL slot (ADVIA 1650/2400)

For storage and stability information, refer to the package insert.

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Calibration
Refer to the package insert supplied with the Siemens Chemistry Calibrator (REF
09784096; T03-1291-62) for handling instructions and values. For setup and use
instructions, refer to the Calibration Overview section of the system-specific Operator’s
Guide.

Calibration Frequency
Perform a calibration when this method is implemented on the system. You must
recalibrate after the following events:
• when the reagent lot number changes
• after replacing critical optical or hydraulic components
• when indicated by quality control procedures

Siemens has validated the calibration stability for this method as shown in the following
table:

System Minimum Calibration Stability*

ADVIA 1200 24 days
ADVIA 1650/1800 24 days
ADVIA 2400 24 days
*or whenever indicated by quality control data

Siemens recommends calibrating new reagent packs if the previous reagent pack was
calibrated any time during its on-board stability, other than as a fresh pack.

Reagent Blank (RBL) Frequency

The RBL is measured at the time of method calibration.

Quality Control
Follow government regulations or accreditation requirements for quality control
frequency.

Siemens recommends the use of commercially available quality control materials with
at least 2 levels (low and high). A satisfactory level of performance is achieved when
the analyte values obtained are within the Acceptable Control Range for the system or
within your range, as determined by an appropriate internal laboratory quality control
scheme.

The actual frequency of control in a laboratory is based on many factors, such as

workflow, system experience, and government regulation. Each laboratory should
evaluate the controls based on the frequency established by their laboratory
guidelines. When the method is performed, analyze at least 2 levels of controls daily.

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Also, assay controls under the following conditions:

• whenever you use a new reagent lot
• following the performance of any system maintenance, cleaning, or troubleshooting
procedure
• after performing a new calibration

For more information, refer to the Quality Control Overview section of the
system-specific Operator’s Guide.

Limitations of the Procedure 2

A number of substances cause physiological changes in serum, plasma, or urine
analyte concentrations. A comprehensive discussion of possible interfering
substances, their serum, plasma, or urine concentrations, and their possible
physiological involvements is beyond the scope of this document. Consult listed
reference for specific details on known potential interfering substances.2

As with any chemical reaction, you must be alert to the possible effect on results of
unknown interferences from medications or endogenous substances. The laboratory
and physician must evaluate all patient results in light of the total clinical status of the
patient.

Interferences
Siemens tested the following potential interferents and found the results shown below:

ADVIA 1200
Ca Sample
Interferent Interferent Level Concentration Interference*
Bilirubin 50 mg/dL 5.9 mg/dL NSI
(855 µmol/L) (1.48 mmol/L)
Hemolysis 1000 mg/dL 5.9 mg/dL NSI
(hemoglobin) (10 g/L) (1.48 mmol/L)
Lipemia (from Intralipid) 1000 mg/dL 5.9 mg/dL NSI
(11.3 mmol/L)** (1.48 mmol/L)
Omniscan† 1.5 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
OptiMARK† 1.0 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
*NSI = No Significant Interference. A percentage effect10% is considered significant
interference.
**as triolein
†magnetic resonance contrast agents

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ADVIA 1650/1800
Ca Sample
Interferent Interferent Level Concentration Interference*
Bilirubin 50 mg/dL 5.9 mg/dL NSI
(855 µmol/L) (1.48 mmol/L)
Hemolysis 1000 mg/dL 5.9 mg/dL NSI
(hemoglobin) (10 g/L) (1.48 mmol/L)
Lipemia (from Intralipid) 1000 mg/dL 5.9 mg/dL NSI
(11.3 mmol/L)** (1.48 mmol/L)
Omniscan† 1.5 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
OptiMARK† 1.0 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
*NSI = No Significant Interference. A percentage effect 10% is considered significant
interference.
**as triolein
†magnetic resonance contrast agents

ADVIA 2400
Ca Sample
Interferent Interferent Level Concentration Interference*
Bilirubin 50 mg/dL 5.9 mg/dL NSI
(855 µmol/L) (1.48 mmol/L)
Hemolysis (hemoglobin) 1000 mg/dL 5.9 mg/dL NSI
(10 g/L) (1.48 mmol/L)
Lipemia (from Intralipid) 1000 mg/dL 5.9 mg/dL NSI
(11.3 mmol/L)** (1.48 mmol/L)
Omniscan† 1.5 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
OptiMARK† 1.0 mmol/L 5.9 mg/dL NSI
(1.48 mmol/L)
*NSI = No Significant Interference. A percentage effect 10% is considered significant
interference.
**as triolein
†magnetic resonance contrast agents

NOTE: Siemens has determined that there is a possibility for certain ADVIA Chemistry
reagents to interact with the CA_2 assay when run on the same system. To mitigate
these carryover events, the ADVIA Chemistry systems software provides a
Contamination Avoidance process. For further information and instructions to establish
this process on your systems, refer to the Customer Bulletin entitled: Consolidated
Directory of Contamination Avoidance Settings for ADVIA Chemistry Systems
(10813375, latest revision).

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Performance Characteristics
Precision 3
Each sample was assayed 2 times per run, 2 runs per day, for at least 10 days.
Precision estimates were computed according to CLSI document EP05-A2, Evaluation
of Precision Performance of Quantitative Measurement Methods; Approved Guideline.3

Data contained in this section represents typical performance for ADVIA Chemistry
systems. Your laboratory data may differ from these values.

Conversion factor: mg/dL x 0.25 = mmol/L

ADVIA 1200
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)

Common Units (mg/dL)

Serum 6.0 0.11 1.8 0.12 2.1
Serum 9.9 0.09 0.9 0.24 2.4
Serum 14.1 0.09 0.6 0.16 1.2
Urine 6.0 0.13 2.1 0.23 3.8
Urine 11.9 0.25 2.1 0.29 2.4
SI Units (mmol/L)
Serum 1.49 0.027 1.8 0.031 2.1
Serum 2.48 0.022 0.9 0.060 2.4
Serum 3.53 0.021 0.6 0.042 1.2
Urine 1.50 0.032 2.1 0.057 3.8
Urine 2.97 0.062 2.1 0.071 2.4

ADVIA 1650/1800
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)
Common Units (mg/dL)
Serum 5.8 0.08 1.4 0.12 2.1
Serum 9.8 0.12 1.3 0.13 1.3
Serum 13.8 0.07 0.5 0.11 0.8
Urine 6.0 0.07 1.2 0.18 3.0
Urine 11.8 0.11 0.9 0.18 1.6
SI Units (mmol/L)
Serum 1.46 0.020 1.4 0.031 2.1
Serum 2.45 0.032 1.3 0.032 1.3
Serum 3.46 0.017 0.5 0.028 0.8
Urine 1.49 0.018 1.2 0.045 3.0
Urine 2.95 0.027 0.9 0.047 1.6

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ADVIA 2400
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)

Common Units (mg/dL)

Serum 6.0 0.12 2.0 0.13 2.2
Serum 9.9 0.19 1.9 0.19 2.0
Serum 14.0 0.11 0.8 0.17 1.2
Urine 6.2 0.14 2.2 0.23 3.8
Urine 11.8 0.13 1.1 0.26 2.2
SI Units (mmol/L)
Serum 1.49 0.030 2.0 0.033 2.2
Serum 2.47 0.047 1.9 0.049 2.0
Serum 3.49 0.028 0.8 0.042 1.2
Urine 1.54 0.034 2.2 0.059 3.8
Urine 2.95 0.032 1.1 0.065 2.2

Analytical Range
This method is linear from 1.0–16.0 mg/dL (0.25–4.00 mmol/L) for serum and plasma,
and 1.0–32.0 mg/dL (0.25–8.00 mmol/L) for urine.

Siemens has validated an automatic rerun condition for this method that extends the
reportable range up to 32.0 mg/dL (8.00 mmol/L) for serum and plasma, and up to
160.0 mg/dL (40.00 mmol/L) for urine.

Expected Values 4,5

The following table lists the reference ranges for this method:

Sample Type Reference Range

Serum/Plasma 4 8.7–10.4 mg/dL
(2.18–2.60 mmol/L)*
Urine5 100–300 mg/day
(2.50–7.50 mmol/day)
Random Urine5 Male 0.9–37.9 mg/dL
(0.225–9.47 mmol/L)
Female 0.5–35.7 mg/dL
(0.125–8.92 mmol/L)

* This range (2.5 and 97.5 percentile values) is based on the determination of serum
calcium in samples from 148 apparently healthy adult volunteers.

Siemens provides this information for reference. Each laboratory should establish its
own normal range. You can enter normal range values and abnormal range values at
the Analytical Parameters (Chemistry) window.

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System Correlation
The performance of the applicable method (y) was compared with the performance of
the same method on the comparison system (x). Clinical specimens and contrived
patient samples were used in the method comparison studies. The studies were
performed at internal and external sites. The number of observations included in
calculations and results are presented in the tables below:

ADVIA 1200
Regression
Specimen Type Comparison System (x) N Equation Sy.x r Sample Range
Serum ADVIA 1650 CA_2 64 y = 0.97x + 0.16 0.16 0.997 3.33–9.9 mg/dL
y = 0.97x + 0.04 0.04 0.997 0.83–2.48 mmol/L
Plasma* ADVIA 1200 (serum) 47 y = 1.02x - 0.19 0.15 0.999 2.28–16.0 mg/dL
y = 1.02x - 0.05 0.04 0.999 0.57–4.00 mmol/L
Urine ADVIA 1650 CA_2 86 y = 0.97x + 0.55 0.41 0.997 1.34–20.5 mg/dL
y = 0.97x + 0.14 0.10 0.997 0.34–5.11 mmol/L
Serum Reference Method 61 y = 0.99x + 0.06 0.26 0.990 4.03–10.3 mg/dL
y = 0.99x + 0.02 0.07 0.990 1.01–2.58 mmol/L
*lithium heparin (matrix comparison)

ADVIA 1650/1800
Regression
Specimen Type Comparison System (x) N Equation Sy.x r Sample Range
Serum ADVIA 1650 CA 170 y = 0.96x + 0.70 0.27 0.996 1.10–15.55 mg/dL
y = 0.96x + 0.18 0.07 0.993 0.28–3.88 mmol/L
Plasma* ADVIA 1650 (serum) 48 y = 0.99x - 0.13 0.34 0.995 2.28–15.3 mg/dL
y = 0.99x - 0.03 0.08 0.995 0.57–3.82 mmol/L
Urine ADVIA 1650 CA 50 y = 0.96x + 0.29 0.34 0.998 1.15–26.75 mg/dL
y = 0.96x + 0.07 0.09 0.998 0.29–6.69 mmol/L
Serum Reference Method 61 y = 1.02x - 0.13 0.20 0.995 4.03–10.3 mg/dL
y = 1.02x - 0.03 0.05 0.995 1.01–2.58 mmol/L
*lithium heparin (matrix comparison)

ADVIA 2400
Regression
Specimen Type Comparison System (x) N Equation Sy.x r Sample Range
Serum ADVIA 1650 CA_2 64 y = 1.00x - 0.01 0.08 0.999 3.33–9.9 mg/dL
y = 1.00x - 0.00 0.02 0.999 0.83–2.48 mmol/L
Plasma* ADVIA 2400 (serum) 47 y = 0.99x - 0.08 0.59 0.984 2.12–16.0 mg/dL
y = 0.99x - 0.02 0.15 0.984 0.53–4.00 mmol/L
Urine ADVIA 1650 CA_2 86 y = 0.98x - 0.10 0.13 0.999 1.34–20.5 mg/dL
y = 0.98x - 0.03 0.03 0.999 0.34–5.11 mmol/L
Serum Reference Method 61 y = 1.02x - 0.15 0.22 0.994 4.03–10.3 mg/dL
y = 1.02x - 0.04 0.06 0.994 1.01–2.58 mmol/L
*lithium heparin (matrix comparison)

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Standardization
The ADVIA CA_2 method is traceable to an Internal Siemens reference method
(Inductively Coupled Plasma Atomic Emission), which uses reference materials from
the National Institute of Standards and Technology (NIST), via patient sample
correlation. See the correlation data in System Correlation for the relationship.
Assigned values of Siemens Chemistry Calibrators are traceable to this
standardization.

References
1. Michaylova V, Illkova P. Anal Chem Acta. 1971;53:194.
2. Young DS. Effects of Drugs on Clinical Laboratory Tests. 3rd ed. Washington:
AACC Press (1990).
3. Clinical and Laboratory Standards Institute (formerly NCCLS). Evaluation of
Precision Performance of Quantitative Measurement Methods; Approved Guideline
- Second Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2004.
NCCLS Document EP05-A2.
4. Data on file.
5. Tietz NW. Clinical Guide to Laboratory Tests. Fourth Edition. St. Louis, MO:
WB Saunders Company; 2006:204.

Technical Assistance
For customer support, please contact your local technical support provider or
distributor.

siemens.com/healthcare

Trademarks
ADVIA is a trademark of Siemens Healthcare Diagnostics.
Intralipid is a trademark of Fresenius Kabi AB.
Omniscan is a trademark of GE HealthCare.
OptiMARK is a trademark of Mallinckrodt Inc.

Made in: UK

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Definition of Symbols
The following symbols may appear on the product labeling:

Symbol Definition Symbol Definition

In vitro diagnostic medical device Catalog number

Authorized Representative in the

Legal manufacturer
European Community

CE Mark with identification number

CE Mark
of notified body
0088

Consult instructions for use Biological risk

Keep away from sunlight and heat Temperature limitation

Lower limit of temperature Upper limit of temperature

Do not freeze (> 0°C) Up

Use by Contains sufficient for (n) tests

Recycle Printed with soy ink

Rev. Revision YYYY-MM-DD Date format (year-month-day)

Batch code

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