The Creative Approach to Bioscience
Calcium SYMBOLS IN PRODUCT LABELLING
Arsenazo III (Single Reagent) EC REP Authorised Representative Use by/Expiration Date
IVD For in-vitro diagnostic use CAUTION. Consult instructions
REF: 227 001 (2 x 30 ml) 60 test LOT Batch Code/Lot number for use
REF: 227 002 (4 x30 ml) 120 test REF Catalogue Number Manufactured by
REF: 227 003 (2 x 100 ml)200 test Consult instructions for use
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Intended Use Specimen Collection and Preservation
Serum and plasma
Spectrum Diagnostics calcium reagent is intended for the in-vitro
quantitative, diagnostic determination of calcium in human serum Use nonhemolyzed serum.Heparin is the only acceptable antico-
on both automated and manual systems. agulant. No other anticoagulant can be used. Fresh serum collected
in the fasting state is the perferred specimen. Serum or plasma
Background should be separated from cells as soon as possible, because
prolonged contact with the clot may cause lower calcium values.
Calcium is the fifth most common element in the body, most of which Sera from patients receiving EDTA (treatment of hypercalcemia) are
(98 %) is present in the skeleton. One half of the remaining calcium unsuitable for analysis, since EDTA will chelate the calcium and
is found in extracellular fluid and the rest in tissues. Calcium has a render it unavailable for reaction with O-cresolphthalein complexone.
crucial role in bone mineralization and is also vital for basic physio- The biological half-life of calcium in blood is few hours.
logical processes such as blood coagulation, neuromuscular
conduction, and normal muscle tone. Calcium is constantly lost from Urine
the body through excretion in feces, urine and to a small extent in
sweat. The determination of serum calcium is useful for monitoring Specimens should be collected in acid washed bottles. 24 hour
myeloma, renal failure, acid base balance,and cirrhosis. Both serum Specimens should be collected in containers containing 5 ml of
and tissue calcium in the body are controlled by parathyroid hormone, 6 mol/L HCl. If the specimen is collected without acid, the pH should
calcitonin and vitamin D. Hypocalcemia may be observed in be adjusted < 3 with 6 mol/L HCl. Dilute urine specimen 2 times with
hypoparathyrodism, steatorrhea, pancreatitis and nephrosis. Increased bidistilled water (1volume urine + 1volume distilled water) before
levels may be associated with multiple myeloma and other neoplastic assay.
diseases.
Stability (serum): 7 days at 15 – 25 oC; 3 weeks at 4 – 8 oC;
Method 8 months at -20 oC
colorimetric. Arsenazo III. Stability (urine): 2 days at 15 – 25 oC; 4 days at 4 – 8 oC;
3 weeks at -20 oC
Assay Principle
Stored serum or urine specimens must be mixed well prior to analysis.
At a neutral pH, the Ca2+ form with Arsenazo III a complex, the
color intensity of which is directly proportional to the concentration System Parameters
of calcium in the sample.
Wavelength 650 nm (600 nm)
Reagents Optical path 1 cm
Assay type End-point
Standard Calcium (ST) Direction Increase
10 mg/dL 2.5 mmol/L Sample : Reagent Ratio 1 : 100
e.g.: Reagent volume 1 ml
Reagent (R) Sample volume 10 µl
MES, pH 6.40 100 mmol/L Temperature 15 - 25 oC
Arsenazo III 200 µmol/L Zero adjustment Reagent Blank
Sensitivity 1 mg/dL (0.25 mmol/L)
Linearity 20 mg/dL (5 mmol/L)
Precautions and Warnings
Do not ingest or inhalate. In case of contact with eyes or skin; rinse
immediately with plenty of soap and water. In case of severe injuries; Procedure
seek medical advice immediately.
Blank Standard Specimen
Reagent Preparation, Storage and Stability Standard …… 10 µl ……
Spectrum Calcium reagents are supplied ready-to-use and stable Specimen …… …… 10 µl
up to the expiry date labeled on the bottles when stored sealed at Reagent 1 ml 1 ml 1 ml
15 – 25 oC.
Mix and incubate for 3 minutes at 20 - 25 oC. Measure absorbance
Deterioration of specimen (Aspecimen) and standard (Astandard) against reagent
blank.
Do not use the Spectrum Calcium reagents if turbid. Failure to recover
control values within the assigned range may be an indication of
reagent deterioration.
Calculation Aspecimen Expected values
Serum calcium concentration (mg/dL) = x 10 Serum, plasma
Astandard
Aspecimen Adults
Urine calcium (mg/24 hrs)= x10 x10*x 2**x V*** 20 - 50 years 8.8-10.2 mg/dl (2.20-2.55 mmol/L)
Astandard >50 years 8.4- 9.7 mg/dl (2.09-2.42 mmol/L)
* The factor “10” converts mg/dl to mg/litre
** The factor “2” represents the dilution factor Children
*** “V” represents the 24-hour urine volume in litres 4 -18years 9.2-11.0 mg/dl (2.30-2.75 mmol/L)
>4 weeks 7.2-11.2 mg/dl (1.80-2.8 mmol/L)
Quality Control
Normal & abnormal control serum of known concentrations should Urine (24 h)
be analyzed with each run. Females <250 mg/day (<6.25 mmol/day)
Males <300 mg/day (<7.5 mmol/day)
Performance Characterstics Childern <6 mg/Kg/day (<0.15 mmol/day)
Precision
Within run (Repeatiblity) Spectrum Diagnostics does not interpret the results of a clinical
laboratory procedure; interpretation of the results is considered
Level 1 Level 2 the responsibility of qualified medical personnel. All indications
of clinical significance are supported by literature references.
n 20 20
Mean (mg/dL) 9.58 13.97 Analytical Range
SD 0.12 0.207 2 – 20 mg/dl (0.5-5 mmol/L).
CV% 1.33 1.48 Waste Disposal
This product is made to be used in professional laboratories.
Run to run (Reproducibility) Please consult local regulations for a correct waste disposal.
S56: dispose of this material and its container at hazardous or
Level 1 Level 2 special waste collection point.
S57: use appropriate container to avoid environmental contamination.
n 20 20 S61: avoid release in environment. refer to special instructions/safety
Mean (mg/dL) 9.62 14.15 data sheets.
SD 0.23 0.221 References
CV% 1.42 1.53 1. Barnett RN: A scheme for the comparison of quantitative methods.
AM J Clin Pathol 43: 562, 1965.
Methods Comparison 2. Fiereck EA: Appendix. Normal values. in:Fundamentals of clinical
A comparison between Spectrum Diagnostics Calcium reagent and chemistry. NW Tietz, editor,Saunders, Philadelphia, p1208,1976.
a commercial reagent of the same methodology was performed on 3. Kessler G, wolfman M: An automated procedure for the
20 human sera. A correlation of 0.979 was obtained. simultaneous determination of calcium and phosphorus.Clin Chem
10:686, 1964.
Sensitivity 4. Peters JP, Van Slyke, DD: Quantitative clinical chemistry, vol
2,williams and wilkins, Baltimor (MD),1932, p 760.
When run as recommended, the minimum detection limit of this 5. Tietz NW: Blood gases and electrolytes. In:Fundamentals of clinical
assay is 2.0 mg/dL. chemistry, NW tietz, editor,Saunders, Philadelphia, 176,pp 903,
908.
Linearity 6. Young DS, Effects of drugs on clinical laboratory tests. AACC
press, Washington, D.C. 1990.
The reaction is linear up to calcium concentration of 20 mg/dl.
Specimens showing higher concentration should be diluted 1+1
using physiological saline and repeat the assay (result • 2). ORDERING INFORMATION
Interfering Substances: CATALOG NO. QUANTITY
Haemolysis 227 001 2 x 30 ml
avoid haemolysis. 227 002 4 x 30 ml
227 003 2 x 100 ml
Icterus
No significant interference.
Lipemia
No significant interference.
Anticoagulants
complexing Anticoagulants such as citrate, oxalate and EDTA must
be avoided.
Egyptian Company for Biotechnology (S.A.E)
Obour city industrial area. block 20008 piece 19 A. Cairo. Egypt.
Tel: +202 4665 1848 - Fax: +202 4665 1847
www.spectrum-diagnostics.com
E-mail:info@spectrum-diagnostics.com
MDSS GmbH
EC REP Schiffgraben 41
30175 Hannover, Germany
IFUFCC56 Rev.(2), 1/1/2007
