3.3.

Phosphorus and Magnesium

Analysis
 Acknowledgements
• Addisa Ababa University
• Jimma University
• Hawassa University
• Haramaya University
• University of Gondar
• American Society for Clinical Pathology
• Center for Disease Control and Prevention-
Ethiopia
 Learning Objectives
Upon completion of this lesson, the student will be able
to:
• Discuss expected phosphorus and magnesium levels
in body fluids based on pathophysiological
responses.

• Describe the principle of analysis of phosphorus and

magnesium in terms of reagents and endpoint
detection.
 Learning Objectives
Upon completion of the lesson the student will be able to:
• Describe specimen requirements for phosphorus and
magnesium analysis.

• Explain the expected reference ranges of phosphorus

and magnesium based on normal physiologic
responses.

• Discuss interpretation of phosphorus and magnesium

results
 Outline
• Introduction
• Source and Clinical Significance
• Methods of Analysis
• Specimen
• Quality Control
• Sources of Error
• Interpretation
• Documentation and Reporting
• Summary
 Terminology
• Hyperphosphatemia: increased phosphate and other
forms of phosphorus in the blood plasma.
• Hypophosphatemia: decreased phosphate and other
forms of phosphorus in the blood plasma.
• Hypermagnesemia: increased magnesium in the
blood plasma.
• Hypermagnesemia: increased magnesium in the
blood plasma.
Introduction to Phosphorus and
Magnesium
Minerals
similar to calcium
Inorganic forms in bone, teeth
Phosphates are intracellular anions
Magnesium is intracellular cation
 Source of Phosphorus and
Magnesium
Diet
Bones: inorganic
Teeth
Intracelluar
Plasma:
protein bound
ionized
• Regulated by kidneys and parathyroid gland
– PTH
– Activated vitamin D (calcitriol)
 Physiologic Functions of P and Mg

Phosphorus/ phosphates
• pH buffering
• Electrolyte balance: intracellular anion
• Shifts internally with Insulin release
Magnesium
• Minor electrolyte: intracellular cation
• Enzyme activator or cofactor
• Minor role in blood hemostasis
 Clinical Significance of Phosphorus
Hyperphosphatemia
Renal failure
Hypophosphatemia:
Primary hyperparathyroidism
Renal tubular acidosis
Fanconi’s syndrome
 Methods of Phosphate Analysis
• Photometric
– Ammonia molybdate method
– Dye-Binding method
 Principles of Phosphate Analysis
Ammonium molybdate Dye-binding
• P + (NH4)6Mo7O24 . 4 H2O • P + Ammonium
 phophomyolybdate molybdate 
complex phophomyolybdate
• UV light absorption complex –(reduction)
• Molybdenum blue
• Absorbs 600 nm
Clinical Significance of Magnesium
• Hypermagnesemia:
– Renal failure
– Increased intake
– Lithium drug excess
– Rate genetic calcium disorders
• Hypomagnesemia:
– Gastrointestinal malabsorption
– Renal diseases such as tubular or glomerular
nephritis
– Drugs
– Hypophosphatemia
 Methods of Magnesium Analysis
• General Principles
– Spectrophotometric Dye-binding
– Atomic Absorption spectroscopy
 Magnesium Analysis
• Spectrophotometric: Metallochromic method
• Mg + Calmagite  colored complex
• Absorbance measured 540 nm
 Magnesium Analysis
Atomic Absorption Spectroscopy
• Reference Method
• Magnesium in sample is excited with unique
wavelength from hollow cathode lamp
supplying radiant energy
• Absorption of light is proportional to
concentration
 Specimens
• Serum (P and Mg)
• Heparinized plasma (magesium)
• Urine (magnesium)
– acidified
 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.
 Sources of Error
• Hemolysis (P and Mg are intracellular)
• Anticoagulated
– EDTA, citrate, heparin falsely decrease for P
– EDTA, citrate and some heparin interferes for Mg
• Not fresh or exposed to heat
• Icterus
• Lipemia
• Detergent in glassware or water (contains P)
 Interpretation of Phosphorus and
Magnesium Results
Phosphorus Reference Ranges:
Adult: 2.5-4.5 mg/dL
Magnesium Reference Ranges:
Adult serum: 1.6-2.6 mg/dL
Urine: 3.0-5.0 mmol/24 hr
Compare patient results with reference ranges
to determine if any results are outside of
normal limits.
 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
 Summary of Phosphorus and
Magnesium Analysis
This lesson emphasized on:
• Source and Clinical Significance of P and Mg
• Methods of Analysis, Specimen, Interpretation
compared to reference ranges
• Quality Control, Sources of Error, and
Documentation and Reporting
 Review Questions
• What is the principle of magnesium analysis
by the colorimetric method?
• What are the specimen requirements for
phosphorus analysis?
• Why should hemolyzed specimens be
avoided?
• Why must urine have an additive for
magnesium analysis?
 Review Question
• What is a common pathological cause of
increased serum phosphorus, magnesium and
potassium in a patient?
 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