®
ACCUCARE
CALCIUM
Arsenazo III
Quantitative determination of Calcium in serum / plasma / Urine Stability
Only for In Vitro Diagnostic use in Serum/Plasma: 3 weeks at 4 – 8°C
8 months at –20°C
ORDER INFORMATION in Urine: 1 day at 20 – 25°C
REF Cont. 4 days at 4 – 8°C
CARZM 50 50 X 1 mL 3 weeks at –20°C
CARZ 100 2 X 50 mL Add 10 mL of concentrated HCl to 24 h urine and heat the
specimen to dissolve calcium oxalate.
Discard contaminated specimens. Freeze only once!
CLINICAL SIGNIFICANCE
Calcium is the most abundant and one of the most important minerals in ASSAY PROCEDURE
the human body. Approximately 99% of body calcium is found in bones.
Operating Instructions
A decrease in albumin level causes a decrease in serum calcium. Low
levels of calcium are found in hypoparathyroidism, Check reagent inventories at least daily to ensure that quantities
pseudohypoparathyroidism, vitamin D deficiency, malnutrition and are sufficient for the planned work load.
intestinal malabsorption. Among causes of hypercalcemia are cancers, Bring all reagents, standard and samples to room temperature 18
large intake of vitamin D, enhanced renal retention, osteoporosis, - 28ºC, prior to analysis.
sarcosidosis, thyrotoxicosis, hyperparathyroidism, multiple myeloma, AUTOMATED PARAMETERS
idiopathic hypercalcemia of infancy, and carcinoma metastasic to bone. Wavelength 650 nm (620 – 650 nm)
Elevated calcium concentration in urine is found in nephrolithiasis and Measurement Against Reagent blank
metabolic acidosis. Reaction Temperature RT (22-30ºC)
Reaction Type End Point
METHOD Reaction Direction Increasing
Photometric test using arsenazo III Incubation 5 Min.
Sample Volume 25 µL
PRINCIPLE Reagent I Volume 1000 µL
Blank Absorbance Limit < 0.80
Calcium with Arsenazo III (1, 8-Dihydroxy-3,6-disulpho-2,7- Units mg/dL
naphthalene-bis (azo)-dibenzenearsonic acid), at neutral pH, yields a
blue colored complex. The intensity of the colour formed is proportional MANUAL ASSAY PROCEDURE
to the calcium concentration in the sample. Pipette into Test Tubes
Blank Standard Test
REAGENT Reagent 1 1000µL 1000µL 1000µL
Reagent 1 : Arsenazo III Reagent
Standard -- 25µL --
Calcium Std : 10 mg/dL
Sample -- -- 25µL
Mix Well & Incubate it for 5 minutes at RT (22-30ºC)
REAGENT PREPARATION
All Reagents are ready to use Read and record absorbance of the reagent blank, standard,
Control and each unknown sample immediately.
CARZ 50 are specially treated monovials with 1ml pre-dispensed
REAGENT STORAGE AND STABILITY
reagent. Just add 25 μl sample / std., Incubate at R. T. for 5 min. &
When stored at recommended storage temperature stated on label,
aspirate. Use the same programme as above.
reagent is stable until the expiration date stated on the bottle and kit box
label
SAMPLE DILUTIONS
This method is linear upto a concentration of 16 mg/dL.
WARNING AND PRECAUTIONS Dilute samples above this concentration 1:1 with DI Water and
For in vitro diagnostic use. Repeat assay. Multiply the result by 2.
Do not use components beyond the expiration date.
Do not mix materials from different kit lot numbers. CALCULATION
Exercise the normal precautions required for handling all
laboratory reagents.
(Ac) Sample
The reagent contains preservative. Do not swallow. Avoid contact
Serum/Plasma = -------------------x 10 (Standard concentration.)
with skin and mucous membranes.
(As) Standard
For detailed information refer Material Safety Data Sheet.
Proceed carefully with this product because due to its nature it can (Ac) Sample
get contaminated easily. Urine 24Hr. = -------------------x 10 x vol. (dL) urine/24 h x 2
Most of the detergents and water softening products used in the (As) Standard
laboratories contain chelating agents. A defective rinsing will
invalidate the procedure. CALIBRATORS AND CONTROLS
For the calibration of automated photometric systems the commercially
WASTE MANAGEMENT available suitable multi-calibrator is recommended.
This method has been standardized against the reference method
Please refer to local legal requirements.
Atomic Absorption Spectrometry (AAS).
It is recommended to run a normal and a pathological control serum
MATERIALS REQUIRED BUT NOT PROVIDED which is commercially available to verify the performance of the
NaCl solution 9 g/L measured procedure. The value of controls should fall within the
General laboratory equipment established limit.
Each laboratory should establish corrective action in case of deviations
SAMPLE COLLECTION AND PRESERVATION in control recovery.
Serum, heparin plasma or urine
Do not use EDTA plasma.
Art No.: IFU/BC/CAL
Effective Date: 20/09/2019
Revision No.: 01
®
ACCUCARE
CALCIUM
Arsenazo III
PERFORMANCE CHARACTERISTICS GLOSSARY OF SYMBOL
WITHIN RUN Consult Instruction for Use Lot Number
Mean
Sample SD CV % Catalog Number Date of Manufacturing
Concentration
Norm 11.25 0.37 3.29% Store between Use By or Expiration Date
Path 16.12 0.51 3.20%
Manufacturer For in vitro Diagnostic use only
RUN TO RUN
Mean Keep away from sunlight Content of the kit
Sample SD CV %
Concentration
Norm 10.50 0.21 2.01% LAB-CARE DIAGNOSTICS (INDIA) PVT. LTD.
Path 16.01 0.44 2.76% C1 Type, Shed No.: 3225, Chemical Zone,
GIDC Sarigam – 396155, Dist. Valsad, Gujarat, India.
Tel.: +91 22 2554 2109 /1558
LINEARITY Email: accucarediagnostics.com; Website: www.labcarediagnostics.com
This method is linear upto a concentration of 16 mg/dL.
Dilute samples above this concentration 1:1 with DI Water and
Repeat assay. Multiply the result by 2.
Limit of detection: The limit of detection for Calcium is 0.04 mg/dl.
METHOD COMPARISON
A comparison of Accucare Calcium with a commercially available assay
(x) using 59 samples gave following results: R2 = 0.9500
REFERENCE VALUES
Serum: 8.8 - 10.2 mg/dl = 2.2 - 2.55 mmol/L
Urine : 100 - 300 mg/24 h = 2.5 - 7.5 mmol/24 h
The reference values are to be considered as indicative only. Every
laboratory should establish its own normal range.
LIMITATION OF THE PROCEDURE
For diagnostic purposes, the results should always be assessed in
conjunction with the patient’s medical history, clinical examination
and other findings.
INTERFERENCE
Bilirubin: No interference found upto 50mg/dl.
Hemoglobin: : No interference found upto 450 mg/dL.
Lipemia: Lipids interferences are possible at 660 nm single
wavelength, try using bi-chromatic wavelength 660/700 nm to
avoid interferences.
These characteristics have been obtained using an automatic
analyzer. Results may vary if a different instrument or a manual
procedure is used.
BIBLIOGRAPHY
1. Bishop, M.L.Dubeb-Von Laufen, J.L., Burtis, Carl Aa and
Ashwood, Titz 110, 61.
2. Burtis, CA and Ashwood, ER (ed.), Tietz Fundamentals of Clinical
Chemistry, 5th edition, W B Saunders Company, Philadelphia,
2001, pp. 797 - 799, 968.
3. 2) Burtis, CA, Ashwood, ER and Bruns, DE (ed.), Tietz Textbook of
Clinical Chemistry and Molecular Diagnostics, 4th edition, Elsevier
Inc., St. Louis, 2006, pp. 1904.
4. 3) Janssen, JW and Helbing, AR, Arsenazo III: An Improvement of
the Routine Calcium Determination in Serum, Eur. J. Clin. Chem.
Clin. Biochem., 29 (3), pp. 197 - 201, 1991.
5. 4) Guder WG, Narayanan S, Wisser H, Zawta B. List of Analytes;
Preanalytical variables. Brochure in: Samples: From Patient to the
Laboratory. GIT Verlag GmbH, Darmstadt, 1996.
6. 5) Young, D.S., Effects of Drugs on Clinical Laboratory Tests, Fifth
Edition, AACC Press, Washington, D.C., 3-149 - 3-158, 2000.
7. 6) Bakker A.J. et. al. Gammopathy interference in clinical
chemistry assays: mechanisms, detection and prevention. Clin
Chem Lab Med 2007; 45(9), 1240 - 1243.
Art No.: IFU/BC/CAL
Effective Date: 20/09/2019
Revision No.: 01
