CALCIUM
For BECKMAN COULTER® AU Series Analyzers
1418-0207 Packaging: 4 x 30 mL
INTENDED USE
Reagent kit for In Vitro quantitative automated measurement by trained laboratory professionals of
the concentration of Calcium in samples of human serum, plasma, or urine, from patients from the
general population. Measurements of Calcium are intended to be used by licensed physicians, along
with other in vitro and in vivo tests and physical examination, as an aid in the clinical assessment of
calcium homeostasis.
This product has been packed for use with BECKMAN COULTER AU series chemistry analyzers
(AU600, AU600IVD, AU400, AU640, AU2700, AU5400, AU480, and AU680).
CLINICAL SIGNIFICANCE
Calcium is an indispensable element for a variety of physiological processes, including, formation,
strength and maintenance of bones and teeth, nerve impulse transmission, muscle contraction,
hormone secretion, cell adhesion and blood coagulation. It is required for the maintenance of cell
membrane permeability, the folding and stabilization of proteins and the regulation of the activity of a
variety of enzymes. It is also a key intracellular messenger, and the most important messenger in
excitable cell signaling. Approximately 99% of total body calcium is stored in bones; the remaining
1% is stored in soft tissues and distributed in the extracellular fluid. Circulating calcium (total serum
calcium) consists of three distinct physicochemical states: 1) metabolically active free calcium ion
~50%; 2) protein bound calcium ~40%; 3) calcium complexed to small anions. Calcium levels are
neither diagnostic nor specific for any particular condition. Hypercalcemia may be caused by several
factors and medical conditions, including hyperparathyroidism, bone and kidney disorders or
increased renal retention, vitamin D intoxication, hereditary conditions, malignancies, endocrine
disorders, granulomatous diseases, milk-alkali syndrome, phosphate and magnesium imbalances,
digestive calcium hyperabsorption, severe dehydration, hemoconcentration, hyperglobulinemia,
medications, overdose of calcium supplements, or may be due to immobility-induced calcium release
from bone tissue. Hypocalcemia may be caused by a variety of conditions, including but not limited
to vitamin-D deficiency or resistance, hypoparathyroidism or pseudoparathyroidism, nutritional
deficiency, malabsorption syndromes, hungry bone syndrome, bone and kidney disorders, acute
inflammation of the pancreas, crash injury, rhabdomyolysis, surgery, phosphate and magnesium
imbalances, medications, and may be due to hypoalbuminemia of any cause. Urinary calcium levels
may be affected by the same conditions that affect blood levels.
METHOD PRINCIPLE
The Arsenazo III colorimetric method is used. In this method, Calcium ions present in the specimen
react with the Arsenazo III dye in slightly acidic solution, to form a stable chelate violet compound,
which absorbs light at wavelengths close to 650 nm. The increase in absorbance is directly
proportional to the concentration of Calcium in the sample.
METHOD LIMITATIONS
Refer to the book “Effects of Preanalytical Variables on Clinical Laboratory Tests” for possible
interference of other pharmaceutical agents in this particular test. Interference of other agents is
described in the “Clinical Guide to Laboratory Tests”.
The reagent is designed especially for use with BECKMAN COULTER AU series chemistry analyzers
(AU600, AU600IVD, AU400, AU640, AU2700, AU5400, AU480, and AU680) analyzers. For chemistry
protocols and further information contact the customer support unit at MEDICON.
REAGENT COMPOSITION
MES buffer (pH 6.5): 100 mM
Arsenazo III: 1.5 mM
Non-reactive ingredients, preservative
WARNINGS – PRECAUTIONS
 This device measures total serum calcium, and does not provide information
regarding the levels of the biologically active free (ionized) calcium.
 In individuals with hypoalbuminemia, low calcium levels may appear due to the
decrease in albumin levels, rather than changes in calcium homeostasis, while
normal calcium levels may be actually pathological.
 Calcium results should be interpreted with caution in critically ill patients, individuals
with end-stage renal disease, neonates, patients receiving transfusions and subjects
with alterations in protein levels, blood pH and temperature. Free (ionized) calcium
is recommended to be measured in these situations.
 This reagent is designed for in vitro diagnostic use. In vitro diagnostic reagents can
be hazardous. They should be handled according to good laboratory techniques.
Avoid inhalation and contact with eyes and skin.
 Samples should be considered as potentially infectious. Handle with special caution.
 The reagent contains Arsenazo III. Avoid swallowing and contact of the reagent with
skin and mucous membranes.
 Never refill empty or half-empty vials; this will cause fast deterioration of freshly
added reagent to the old one.
 Dispose all waste according to national laws.
 SDS is available by MEDICON.
Any serious incident that may occur in relation to this device must be reported by
the user to the manufacturer and the competent authority of the country in which
the user and/or the patient is established.
PREPARATION
Reagents are liquid and ready for use. Vials bear barcode for recognition by BECKMAN
COULTER AU series chemistry analyzers (AU600, AU600IVD, AU400, AU640, AU2700,
AU5400, AU480, and AU680).
REAGENT DETERIORATION
The reagent should not be used:
 When it does not exhibit the specified linearity or control values lie outside the
acceptable range after recalibration.
 When it appears turbid or decolorized.
 After prolonged exposure to sunlight or high temperature.
BECKMAN COULTER® AU-series: Registered Trademark of Beckman Coulter, Inc., 250 South Kraemer Boulevard, Brea CA 92821-6232, USA.
MEDICON HELLAS S.A. - Melitona 5-7, 153 44 Gerakas, Greece. Tel: +302106606000 - Fax: +302106612666 – www.mediconsa.com
General Electronic Commercial Registry (G.E.MI.) No. 414401000
MEDICON HELLAS S.A. reserves the right to change the information contained in the insert without prior notice.
Ν-DOC-RA-001A-ver01-rev09-26.05.2023-en
2000
SHELF LIFE
Unopened, reagents are stable up to the stated expiry date when stored at 2 – 8°C. Do
not freeze. Freezing may destroy this device. Reagents loaded on the cooled analyzer tray
are stable for 2 months.
SAMPLE
Use established Good Laboratory Practices for sampling, transport, and separation from
blood cells. Do not use hemolyzed, contaminated or turbid sample specimens. Calcium is
measured in serum, urine, and heparinized plasma. Calcium is stable in serum or plasma
for 7 days at room temperature, for 3 weeks at 4 – 8°C, and for 8 months at -20°C. Calcium
is stable in urine for 2 days at room temperature, 4 days at 4-8°C, and more than 3 weeks
at -20°C (Guder et al, 2001)
Note: Changes in posture may cause fluid shifts that affect total calcium levels. Standing
and upright-posture may cause increases of the total calcium concentration, whilst
recumbency has the opposite effect. Incorrect or prolonged application of tourniquet and
intense exercise can affect the results. When a tourniquet must be used, it should be
applied only just before sampling and released the earlier possible.
Urine samples should be acidified with 10 ml of 6N HCl, added to the collection tube before
collection begins. (Tietz, 1995)
CALIBRATION MEDICON provides MEDI-CAL (1578-0891) traceable to NIST reference material
SRM 956b and SRM909b and to the Atomic Absorption Reference Method for serum calibration.
Calibrate the assay upon first use. Recalibrate every 2 months. Calibration should be repeated for
every new reagent lot or when a significant shift in control values occurs, after major maintenance is
performed on the analyzer or after a critical part is replaced. Urine application protocols are pre-
programmed on the analyzer to automatically acquire a calibration factor after a successful serum
calibration.
QUALITY CONTROL MEDICON provides the MEDICON Clinical Chemistry Control Level 1 & 2
(1578-0901-12 & 1578-0902-12 respectively) for serum quality control. Any commercially available
quality control can be used for other types of samples. Control recovery should lie within the
acceptable range. Results outside the acceptable range even after recalibration could be due to
reagent deterioration, unsuitable storage conditions of control material or control deterioration,
instrument malfunction, or error during test procedure.
MATERIALS REQUIRED BUT NOT PROVIDED WITH THE KIT
BECKMAN COULTER AU series chemistry analyzers (AU600, AU600IVD, AU400, AU640, AU2700,
AU5400, AU480, and AU680) analyzer
Calcium Calibrator
Quality control material
Common laboratory equipment
REFERENCE INTERVALS
Serum: 8.4 – 10.3 mg/dL
Urine 24 hours: 100 – 300 mg/24 hours
In a low calcium diet, reference intervals for urine are: 50 – 150 mg/ 24 hours
In a no calcium diet, reference intervals for urine are: 5 – 40 mg/ 24 hours (Tietz, 1995)
Expected values may vary with age, sex, sample type, diet, and geographical location. Each
laboratory should determine its own expected values as dictated by good laboratory practices.
Calcium results are intended to be used by licensed physicians, along with other in vitro and in vivo
tests and physical examination, as an aid in the clinical assessment of calcium homeostasis. In this
sense, calcium results cannot be used alone for diagnosis, and an affected population cannot be
defined, based on calcium values. All results outside the reference interval need to be evaluated in
view of the other factors mentioned for an informed medical decision.
WASTE DISPOSAL
Dispose of contents/container in accordance with licensed collector’s sorting instructions.
SPECIFIC PERFORMANCE CHARACTERISTICS
The following values are representative of the reagent performance achieved on BECKMAN
COULTER AU series chemistry analyzers (AU600, AU600IVD, AU400, AU640, AU2700, AU5400,
AU480, and AU680) in our own laboratory. Values largely depend on the working conditions and
service level of the analyzer, so they may be different to those that can be obtained in each individual
lab.
Measuring Range:
Serum: 0.01 – 15 mg/dL
Urine: 0.08 – 35 mg/dL
The measuring range is the concentration interval between LoQ and upper limit of linearity. When
values exceed this range samples should be diluted accordingly.
Note: Confirmatory experiments in our laboratory yielded linearity up to 17 mg/dL for serum and 38
mg/dL for urine.
Limit of Detection:
Serum: 0.01 mg/dL
Urine: 0.08 mg/dL
The limit of detection (LoD) is defined as the lowest concentration which can be measured with
acceptable precision. The LoD was determined using a method in accordance with the CLSI-EP17A
procedure.
This is an observational study only, with no specific acceptance criteria. However, it is reasonable to
expect that the LoD will be similar to the lowest linearity limit for each assay method and sample type
tested.
Precision: Within-run studies are performed at 3 different concentration levels (low, mid, and high
levels) using 20 repeats per level. The reproducibility of the reagent is assessed as total precision
following CLSI EP5-A3 guidelines on serum and urine specimens. Within-run %CV should be lower
than 5% in all levels tested for all lots. Total precision %CV should be lower than 8% in all levels
tested for all lots.
Serum Urine
Level (mg/dL) %CV Level (mg/dL) %CV
7.0 3.45 5.9 4.15
9.4 1.21 10.9 1.02
15.6 0.94 19.2 0.92
Level (mg/dL) Total CV Level (mg/dL) Total CV
7.5 5.92 6.2 5.84
9.5 5.28 10.3 2.95
14.8 5.78 16.8 2.48
Analytical Specificity
Interferences - Criterion: recovery within ±10% from target value
Serum (Insignificant up to) Urine (Insignificant up to)
Bilirubin 20 mg/dL Conj. Bilirubin 40 mg/dL
Hemoglobin 500 mg/dL Hemoglobin 500 mg/dL
Triglycerides 2700 mg/dL Magnesium 4 mmol/L
Magnesium 4 mmol/L Ascorbic acid 50 mg/dL
Ascorbic acid 3 mg/dL Glucose 2 g/dL
Albumin 500 mg/dL
IgG 500 mg/dL
Boric acid 500 mg/dL
pH 2–6
Specific Gravity 1.010 – 1.030
Refer to Young DS. (1997) for further information on interfering substances
BIBLIOGRAPHY
1. Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt,
Germany: GIT Verlag; 2001:30–1]
2. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. 2nd. ed. Washington,
DC: The American Association for Clinical Chemistry Press, 1997.
3. Tietz, N.W. (Ed.), Clinical Guide to Laboratory Tests, 3rd Edition, W.B. Saunders Co.,
Philadelphia, PA (1995)
4. Victor W. Rodwell; David Bender; Kathleen M. Botham (22 June 2018). Harper's Illustrated
Biochemistry Thirty-First Edition. McGraw-Hill Education.
5. Nelson David L.; Cox Michael M. (2005). Lehninger Principles of Biochemistry. W. H. Freeman.
6. Carl A. Burtis; Edward R. Ashwood; David E. Bruns. Tietz Textbook of Clinical Chemistry and
Molecular Diagnostics. Elsevier Health Sciences.
7. Richard A. McPherson; Matthew R. Pincus (5 April 2017). Henry's Clinical Diagnosis and
Management by Laboratory Methods. Elsevier Health Sciences.
8. Peacock M. Calcium Metabolism in Health and Disease. CJASN January 2010, 5 (Supplement 1)
S23-S30;
9. Blaine J, et al. Renal control of calcium, phosphate, and magnesium homeostasis. Clin J Am Soc
Nephrol. 2015 Jul 7;10(7):1257-72.
10. Labtest online | AACC. Calcium. Available at: https://labtestsonline.org/tests/calcium Assessed
[29 July 2020]
11. Geusens, P., Sambrook, P. N., & Lindsay, R. (2004). Osteoporosis in Clinical Practice.
doi:10.1007/978-0-85729-402-9
12. AACC. To Correct or Not to Correct: pH-Adjusted Free Calcium Available at:
https://www.aacc.org/community/aacc-academy/publications/scientific-shorts/2012/to-correct-or-
not-to-correct-ph-adjusted-free-calcium. Assessed [29 July 2020]
13. Association of clinical biochemistry and laboratory medicine. Calcium monograph. Available at:
http://www.acb.org.uk/Nat%20Lab%20Med%20Hbk/Calcium.pdf Assessed [29 July 2020]
SYMBOLS

BECKMAN COULTER® AU-series: Registered Trademark of Beckman Coulter, Inc., 250 South Kraemer Boulevard, Brea CA 92821-6232, USA.

MEDICON HELLAS S.A. - Melitona 5-7, 153 44 Gerakas, Greece. Tel: +302106606000 - Fax: +302106612666 – www.mediconsa.com
General Electronic Commercial Registry (G.E.MI.) No. 414401000
MEDICON HELLAS S.A. reserves the right to change the information contained in the insert without prior notice.
Ν-DOC-RA-001A-ver01-rev09-26.05.2023-en