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The Determination of Calcium and Magnesium in Blood Serum and Urine

Applications | 2010 | Agilent TechnologiesInstrumentation
AAS
Industries
Clinical Research
Manufacturer
Agilent Technologies

Summary

Significance of the Topic


Determining calcium and magnesium levels in blood serum and urine is critical for clinical diagnostics, nutritional studies, and monitoring metabolic disorders. Atomic absorption spectrophotometry offers a sensitive and specific approach to quantify these electrolytes, enabling rapid, routine analysis in hospital and research laboratories.

Objectives and Study Overview


This application note describes a validated method for simultaneous determination of calcium and magnesium in biological fluids. The protocol aims to suppress interferences, achieve accurate quantification across relevant concentration ranges, and provide guidelines for routine clinical and industrial testing.

Methodology and Instrumentation


Sample Preparation:
  • Blood serum is diluted 1:10 to 1:20 with aqueous solution containing an ionization suppressant (lanthanum or strontium chloride) or EDTA to mitigate protein and phosphate interferences.
  • Urine is diluted 1:50 to 1:100 with deionized water; simple dilution often suffices, though deproteinization with 10% trichloroacetic acid can improve accuracy for serum.

Analytical Conditions:
  • Calcium is measured at the 422.7 nm resonance line using a nitrous oxide–acetylene flame for optimal sensitivity; air–acetylene may be substituted with a threefold sensitivity loss.
  • Magnesium is measured at the 285.2 nm resonance line in an air–acetylene flame; nitrous oxide–acetylene is also applicable if an appropriate ionization suppressant is present.
  • Standard solutions contain 1.0–3.0 ppm Ca and 0.2–0.6 ppm Mg with 5000 ppm lanthanum or strontium as blank and suppressant.

Used Instrumentation


Techtron Model 1000 atomic absorption spectrophotometer with R-213 photomultiplier.
Operational parameters:
  • Lamp current: 4 mA for Ca, 3 mA for Mg
  • Spectral bandpass: 2 Å
  • Burner: AB-50, 5 cm for Ca; AB-51, 10 cm for Mg

Main Results and Discussion


Calibration graphs for Ca and Mg showed high linearity over the tested range. The presence of lanthanum effectively suppressed ionization and phosphate interferences, ensuring accurate readings. Use of burner rotation and higher concentration standards extends the dynamic range when analyte levels exceed routine calibration.

Benefits and Practical Applications of the Method


  • Simple sample preparation and rapid analysis allow high throughput in clinical and QA/QC laboratories.
  • High sensitivity and selectivity yield precise quantification of trace to mid-level concentrations.
  • Flexibility of flame selection and suppressant usage accommodates various matrix interferences.

Future Trends and Applications


Advancements may include automated sample handling, integration with chromatography for speciation studies, miniaturized flame cells, and coupling with inductively coupled plasma spectrometry for broader elemental coverage. Improved matrix correction algorithms and novel suppressants will further enhance accuracy in complex biological samples.

Conclusion


Atomic absorption spectrophotometry provides a robust, accurate, and efficient method for routine determination of calcium and magnesium in serum and urine. With minimal sample handling and reliable interference control, the described protocol meets clinical and research demands for electrolyte analysis.

References


  1. Willis JB. Nature. 1960;186:249.
  2. Willis JB. Spectrochim Acta. 1960;16:259.
  3. Willis JB. Spectrochim Acta. 1960;16:273.
  4. Willis JB. Spectrochim Acta. 1960;16:551.
  5. Dawson JB, Heaton FW. Biochem J. 1961;80:99.
  6. Willis JB. Anal Chem. 1961;33:556.
  7. Stewart WK, Hutchinson F, Fleming LW. J Lab Clin Med. 1963;61:858.
  8. Zettner A, Seligson D. Clin Res. 1963;11:406.
  9. Zettner A, Seligson D. Clin Chem. 1964;10:869.
  10. Gimblet EG, Marney AF, Bonsnes RW. Clin Chem. 1967;13:204.
  11. Iida C, Fuwa K, Wacker W. Anal Biochem. 1967;18:18.
  12. Pybus J. Clin Chim Acta. 1969;23:309.
  13. Hunt BJ. Clin Chem. 1969;15:979.

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