3.2.

Calcium Analysis

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 Learning Objectives
Upon completion of this lesson, the student will be able
to:
• Discuss expected total calcium and ionized calcium
levels in body fluids based on pathophysiological
responses.

• Describe the principle of analysis of total calcium and

ionized calcium in terms of electronic components,
reagents and endpoint detection.

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 Learning Objectives
Upon completion of the lesson the student will be able
to:

• Describe specimen requirements for calcium

analysis.

• Explain the expected reference ranges of calcium

and ionized calcium based on normal physiologic
responses.

• Discuss interpretation of calcium results

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 Outline
• Introduction
• Source and Clinical Significance
• Methods of Analysis
• Specimen
• Interpretation
• Quality Control
• Sources of Error
• Documentation and Reporting
• Summary

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 Introduction to Calcium
• Minor electrolyte – mineral
• Diet origin
• Extracellular electrolytes
• Bound to protein
• Ionized forms
• Intracellular functions
• Role in bone and kidney control

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 Terminology

Hypercalcemia: increased calcium levels in

plasma
Hypocalcemia: decreased calcium levels in
plasma

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 Source of Calcium
Bones
Teeth
Plasma and Extracellular Fluid
50% plasma calcium is Ca++
50% plasma calcium protein-bound

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 Physiologic Functions of Calcium

• Intracellular Ca++
– Muscle contraction
– Metabolism
– Hormone secretion
– Hemostasis/blood clotting
– Enzyme activation
– Nerve conduction
• Extracellular Ca++
– Electrolyte balance
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 Physiologic Control of Calcium
• Parathyroid hormone (PTH)
– Homeostasis of Calcium and P
• Calcitonin
• Active Vitamin D

• Renal Tubular
– Reabsorb Calcium (into plasma)
– Exchange P (into urine)

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 Clinical Significance of Calcium (total)
• Hypercalcemia
– Primary hyperparathyroidism
– Cancers
– Kidney stones
– Sarcoidosis
• Hypocalcemia
-The most common cause of low total serum calcium
is hypoalbuminemia
– Secondary hyperparathyroidism
– Renal failure
– GI loss

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Clinical Significance of ionized Calcium
• A more specific test for determining active calcium status
compared to total calcium and especially useful for
detecting:
• Hypercalcemia due to:
– Primary hyperparathyroidism
• Hypocalcemia due to:
– Secondary hyperparathyroidism
– Renal failure or GI loss
– Premature infants

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 Methods of Calcium Analysis
• General Principles of Total Calcium
– Spectrophotometric
– Titrimetric
– Atomic Absorption spectroscopy
• General Principles of Ionized Calcium
– Electrochemistry

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 Spectrophotometric Calcium
Analysis
Total Calcium + acid
free calcium
Free Calcium + O-
cresolphthalein
complexone red
chromagen
Measured at 580 nm

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 Calcium Analysis
• Atomic Absorption Spectroscopy
• Reference Method
• Calcium in sample is excited with unique
wavelength from hollow cathode lamp
supplying radiant energy
• Absorption of light is proportional to
concentration

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 Ionized Calcium Analysis
• Ion Selective Electrode:
• Electrode with PVC
membrane selective for Ca+
+

• Ag/ AgCl reference

electrode has constant
voltage potential
•  potential in the
reference solution is
measured due to presence
of Ca++

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 Specimens for Calcium
• Serum
• Heparinized plasma
• Urine, acidified
– Random
– 24 hour urine

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 Interpretation of Calcium Results
Reference Ranges:
• Adult serum total calcium: 8.6-10.2 mg/dL
• Urine calcium: 50-150 mg / 24 hr (dietary dependent)
• Adult plasma Ca++: 1.15 to 1.33 mmol/L
• Compare patient results with reference ranges to
interpret calcium results for hyper- or hypocalcemia.
• Compare patient results with reference ranges to
determine if any results are outside of normal limits.

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 Quality Control
• A normal & abnormal quality control samples
should be analyzed along with patient samples,
using Westgard or other quality control rules for
acceptance or rejection of the analytical run.
– Assayed known samples
– Commercially manufactured

• Validate patient results

• Detects analytical errors.
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 Quality Control Monitoring for
Analytic Errors
• QC performed daily &
following calibration
• Plot on chart
• Follow QC rules
• Determine if current
patient results are
acceptable

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 Sources of Error
• Wrong anticoagulant
• Hemolysis
• Lipemia
• Icterus
• Unpreserved urine
• Reagent deterioration
• Instrument temperature fluctuation
• Nonlinearity of reaction
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 Reporting and Documentation
• To avoid post-analytic errors,
• Report the patient result with :
– Right name and result
– Include reference ranges
– Timely manner

• QC and patient results should be documented

in logbook and retained in lab
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 Summary of Calcium Analysis
This lesson emphasized on:
• Source and Clinical Significance of total and
ionized calcium
• Methods of Analysis, Specimen, Interpretation
compared to reference ranges
• Quality Control, Sources of Error, and
Documentation and Reporting

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 References
• W.Tietz, Phd, et al. Electrolytes and Blood Gases, In
: Fundamental of Clinical Chemistry 5th ed 2001 by
Saunders.
• W Arneson, J Brickell. Clinical Chemistry: A
Laboratory Perspective. 1st Ed. 2007 FA Davis

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