How do you measurement calcium and phosphate using a spectrophotometer?

Answer 1

The appearance of kits for the determination of micro amounts of calcium stimulated

our interest in the use of Eriochrome Blue SE for this analysis. A study of spectrophotometric

curves indicates that, at pH values above 13.7, calcium will complex

and cause a change of dye absorbance while magnesium does not complex with the

dye. A differential spectrophotometric technic is described in this paper in which

the spectrophotometer is set at zero absorbance with a dye-calcium standard solution

or a dye-calcium sample solution as reference and the absorbance of the dye

solution then measured. For a set of standards the absorbance-calcium relationship is

linear. With the Beckman spectrophotometer, Model Dli, it has been found that 1

part of serum to 100 parts of alkaline dye solution (100 A of serum to 10 ml. alkaline

dye solution) can be used. The technic shows greater sensitivity and accuracy than

do previous methods using Eriochrome Blue SE.

Answer 2

To measure calcium and phosphate using a spectrophotometer, you would need to use specific reagents that can form colored complexes with these ions. For calcium, you can use a chromophore such as o-cresolphthalein complexone, which forms a purple complex in the presence of calcium ions. For phosphate, you can use reagents such as ammonium molybdate and ascorbic acid, which form a blue complex with phosphate ions. By measuring the absorbance of these colored complexes at specific wavelengths using a spectrophotometer, you can quantify the concentrations of calcium and phosphate in your sample.