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MLAB 2401: Clinical Chemistry Keri Brophy -Martinez

MLAB 2401: Clinical Chemistry Keri Brophy -Martinez. Methods of Measurement: Mineral and Bone Metabolism. Determination of Total Calcium. Colorimetric O-cresolphthalein Calcium in alkaline solution reacts with o-cresolphthalein complexone to form a purple complex

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MLAB 2401: Clinical Chemistry Keri Brophy -Martinez

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  1. MLAB 2401: Clinical ChemistryKeri Brophy-Martinez Methods of Measurement: Mineral and Bone Metabolism

  2. Determination of Total Calcium • Colorimetric • O-cresolphthalein • Calcium in alkaline solution reacts with o-cresolphthalein complexone to form a purple complex • Proportional to the calcium concentration • Uses 8-quinolinol or 8-hydroxyquioline to remove Mg ions • Arsenzo III dye • Atomic Absorption Spectrophotometry • Reference method

  3. Specimen Requirements:Calcium( total) • Serum • Lithium heparin plasma • Avoid the use of anticoagulants with EDTA or oxalate

  4. Determination of Free Calcium • Ion selective electrodes used • pH dependent • Selectively and reversibly bind calcium ions • As calcium binds to the membrane a potential develops which is proportional to the concentration

  5. Specimen Requirements:Calcium (Ionized) • Samples must be collected anaerobically • Heparinized whole blood preferred • Plasma • Serum

  6. Reference Ranges • Calcium, total • 8.60-10.0 mg/dL • Ionized Calcium • 4.60-5.08 mg/dL

  7. Determination of Phosphorus • Formation of ammonium phosphomolybdate complex • Complex measured by UV absorption at 340 nm or reduced to molybdenum blue and read between 600-700 nm

  8. Specimen Requirements: Phosphorus • Serum • Lithium heparin plasma • Avoid hemolysis and certain anticoagulants • Diural variation a factor

  9. Reference Range: Phosphorus • 2.5-4.5 mg/dL

  10. Determination of Magnesium • Colorimetric • Calmagite • Mg2+ binds with calmagite to for a reddish-violet complex, read at 532 nm • Formazen dye • Mg2+ binds with the dye to form a colored product read at 660 nm • Methylthymol blue • Mg2+ binds with the chromogen to form a colored complex • Atomic Absorption Spectrophotometry • Reference method

  11. Specimen Requirements: Magnesium • Serum • Heparinized plasma • Avoid hemolysis • Remove serum from cells ASAP

  12. Reference Range: Magnesium • 1.6-2.6 mg/dL

  13. References • Bishop, M., Fody, E., & Schoeff, l. (2010). Clinical Chemistry: Techniques, principles, Correlations. Baltimore: Wolters Kluwer Lippincott Williams & Wilkins. • Sunheimer, R., & Graves, L. (2010). Clinical Laboratory Chemistry. Upper Saddle River: Pearson .

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