Determination of Total Calcium and Magnesium Ion Concentration

Safety

Lab coats, safety glasses and enclosed footwear must be worn at all times in the laboratory.
Concentrated ammonia solution used in preparing the buffer and indicator solutions is highly
corrosive − wear rubber gloves and take care when handling. Both the buffer and indicator
(and thus also the titration solution) will liberate ammonia gas to some extent. This gas may
be harmful if inhaled in large quantities. Work in a fume hood or well-ventilated area.

Introduction

This method, called a complexometric titration, is used to find the total calcium and
magnesium content of milk, sea water and various solid materials. It can also be used to
determine the total hardness of fresh water provided the solutions used are diluted. The
combined concentration of calcium and magnesium ions is considered to be the measure of
water hardness. The method uses a very large molecule called EDTA which forms a
complex with calcium and magnesium ions. EDTA is short for ethylenediaminetetraacetic
acid. A blue dye called Eriochrome Black T (ErioT) is used as the indicator. This blue dye
also forms a complex with the calcium and magnesium ions, changing colour from blue to
pink in the process. The dye–metal ion complex is less stable than the EDTA–metal ion
complex. For the titration, the sample solution containing the calcium and magnesium ions is
reacted with an excess of EDTA. The indicator is added and remains blue as all the Ca2+
and Mg2+ ions present are complexed with the EDTA. A back titration is carried out using a
solution of magnesium chloride. This forms a complex with the excess EDTA molecules until
the end-point, when all the excess EDTA has been complexed. The remaining magnesium
ions of the magnesium chloride solution then start to complex with ErioT indicator,
immediately changing its colour from blue to pink. The main reaction is

Ca2+ + EDTA4- → [Ca-EDTA]2-

Back titration

EDTA4- + Mg2+ → [Mg-EDTA]2-

Indicator reaction: note, ErioT is blue and ErioT-Mg is pink

ErioT + Mg2+ → ErioT-Mg

1
Equipment Needed

Burette

20 mL pipette

250 mL conical flasks

100 mL volumetric cylinder

Solutions Needed

EDTA: (ethylenediaminetetraaceticacid) 500 mL of a 0.05 molL-1 solution. Weigh 9.31 g of

the EDTA salt and dissolve it in 500 mL of distilled water in a volumetric flask.

Buffer: Dissolve 7.0 g of ammonium chloride in 57 mL concentrated ammonia (see safety

notes). Dilute to 100mL with distilled water in a volumetric flask. The pH should be 10.5.

MgCl2 .6H2O: 0.025 molL-1 solution. Weigh 2.54 g of magnesium chloride hexahydrate and
dilute to 500 mL with distilled water in a volumetric flask.

ErioT indicator: Dissolve 0.2 g of Eriochrome Black T indicator in 15 mL of concentrated

ammonia solution (or 15 mL of triethanolamine) (see safety notes) and 5mL absolute
ethanol.

Method

Sample Preparation

For samples that are already in solution, such as freshwater, seawater and milk, no further
preparation is needed. For solid samples such as eggshells and limestone, the samples
must first be dissolved in acid. Accurately weigh about 0.5 g of the solid into a small beaker
or conical flask, add about 20 mL dilute hydrochloric acid and allow the solid to completely
dissolve (this may take several minutes). Neutralise the unreacted acid with dilute sodium
hydroxide solution until the pH of the solution is almost 7 (according to pH indicator paper).
For eggshells, the inner membrane will remain undissolved and may be carefully removed
from the solution. Transfer the solution to a 100 mL volumetric flask and make up to the
mark with distilled water.

Standardisation of the EDTA Solution

1. Pipette a 10 mL sample of the EDTA solution into a conical flask.

2. Add 10 mL of ammonia buffer solution and 1 mL of Eriochrome Black T indicator solution.

2
3. Titrate the EDTA with the magnesium chloride solution until the endpoint is reached – a
permanent colour change from blue to pink.

4. Having determined the average titre of the magnesium chloride solution, determine the
number of moles used. 5. Given the Mg2+ : EDTA ratio of 1:1, calculate the concentration of
your EDTA solution.

5. Given the Mg2+ : EDTA ratio of 1:1, calculate the concentration of your EDTA solution.

Titration Method for Milk

1. Pipette 10 mL of the sample solution into a conical flask.

2. Add 20 mL of 0.05 mol L−1 EDTA solution.

3. Add 10 mL of ammonia buffer, 50 mL of distilled water and 1 mL of Eriochrome Black T

indicator solution.

4. Titrate the sample with the standard 0.025 molL−1 magnesium chloride solution until a
permanent pink colour appears

Titration Method for Fresh or Tap Water Samples

1. Add a 100 mL of the sample solution into a 250 mL conical flask.

2. Prepare a 0.005 mol L−1 EDTA solution by diluting the 0.05 mol L−1 EDTA solution by a
factor of 1/10. Add 20mL of this diluted EDTA to the sample solution.

3. Add 10 mL of the ammonia buffer and 1 mL of Eriochrome Black T indicator solution.

3
 Figure 2 Same colour changes for magnesium chloride
back-titration as in Figure 1, but for cloudy (opaque)
sample solution, eg milk. Left flask: blue colour well
before endpoint. Centre flask: last trace of blue/purple.
Right flask: pink/red colour at endpoint

Figure 1 Colour changes for magnesium chloride

back-titration in clear solution using Eriochrome Black
T indicator. Left flask: blue colour well before endpoint
(all Ca2+/Mg2+ ions complexed by excess EDTA, all
indicator molecules uncomplexed). Centre flask: last
trace of blue/purple colour just before endpoint
(excess EDTA almost totally complexed by added
Mg2+). Right flask: pink/red colour at endpoint (all
EDTA complexed by added Mg2+, indicator also
complexed).
4. Prepare a 0.0025 mol L−1 magnesium chloride solution by diluting the 0.025 mol L−1
magnesium chloride solution by a factor of 1/10.

5. Titrate the sample solution with this 0.0025 molL−1 magnesium chloride solution until a
permanent pink colour appears. Repeat the titration with further samples until concordant
results (titres agreeing within 0.1 mL) are obtained.

Result Calculations

1. Calculate the total moles of EDTA added to the sample solution.

2. Calculate the moles of the magnesium chloride solution used in the back titration from
your concordant results. From the equation of the titration below, the moles of Mg2+ will
be equivalent to the moles of excess EDTA.

EDTA4- + Mg2+ → [Mg-EDTA]2-

3. Given the ratio of Ca2+ + Mg2+ : EDTA = 1 : 1, calculate the moles of Ca2+ and Mg2+ that
must have been complexed with EDTA by subtracting the excess EDTA from the total moles
of EDTA added to the sample. These results are the moles of Ca2+ and Mg2+ in the
sample solution.

4
5