College of Science
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 fumehood 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 dyemetal
ion complex is less stable than the EDTAmetal
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
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. Do not store more than one to
two days before use. You may be able to get the ErioT
indicator from the University of Canterbury - see the
contact details at the end.
Method Titration Method for Seawater, Milk and Solid
Samples
Sample Preparation 1. Pipette 10 mL of the sample solution into a conical
For samples that are already in solution, such as flask.
freshwater, seawater and milk, no further preparation 2. Add 20 mL of 0.05 mol L1 EDTA solution.
is needed.
3. Add 10 mL of ammonia buffer, 50 mL of distilled
For solid samples such as eggshells and limestone, water and 1 mL of Eriochrome Black T indicator
the samples must first be dissolved in acid. Accurately solution.
weigh about 0.5 g of the solid into a small beaker or
conical flask, add about 20 mL dilute hydrochloric acid 4. Titrate the sample with the standard 0.025 molL1
and allow the solid to completely dissolve (this may magnesium chloride solution until a permanent pink
take several minutes). Neutralise the unreacted acid colour appears.
with dilute sodium hydroxide solution until the pH Titration Method for Fresh or Tap Water Samples
Eriochrome Black T indicator solution.
Eriochrome Black T indicator solution. 4. Prepare a 0.0025 mol L 4. Preparechloride
magnesium a 0.0025
1 mol L magnesium chloride
1

of3. the solution is almost 7 (according to pH

solution indicator
by diluting the 0.025 solution
mol L by diluting
magnesium the
1 0.025 mol L magnesium 1
Titrate the EDTA with the3.magnesium
Titrate the EDTA with the magnesium chloride
chloride 1. 1/10.Add a 100 by amL ofofthe
1/10. sample solution into a 250 mL
paper). Fortheeggshells,
solution until the inner
endpoint issolution
reached until themembrane will
chloride
endpoint is reached
a permanent remain
solution
a permanentby a factor ofchloride solution factor
colour change from blue may colour change from blue to pink.
to pink. 5. Titrate 5. with conical
Titrate
thisthe flask.
samplemolLsolution with this 0.0025 molL 1
fromthe sample solution 0.0025 1
undissolved and be carefully removed magnesium
the
chloride solution magnesium
until a chloride
permanent solution until a permanent
pink pink
4. Having determined the average
solution. Transfer the 4. Having
solution determined
titre of the the average titre of the
to asolution,
100 mL volumetric 2. Prepare
colour appears. asamples
Repeat0.005 mol with
the titration L1 EDTA solution by diluting the
further samples
magnesium chloride solution,magnesium
determine the chloride
number of colour appears.
determine Repeat the
the number of titration with further
flask and make up tomoles
moles used. theused. mark with distilled until concordant
water. results until
(titres 0.05 mol L EDTA solution by a factor of 1/10. Add
concordant
agreeing within 0.1 1 (titres agreeing within 0.1 mL)
results
mL)
are obtained. are obtained.
5. Given the Mg : EDTA ratio 5. of Given
2+ the Mg the
1 : 1, calculate : EDTA ratio of 1 : 1, calculate the
2+ 20mL of this diluted EDTA to the sample solution.
Standardisation
concentration of your EDTA ofsolution.
the EDTAofSolution
concentration your EDTA solution.
Result Calculations Result 3. AddCalculations
10 mL of the ammonia buffer and 1 mL of
1. Titration
Pipette a 10formL
Method sample
Seawater, Milkof
Titration andthe
Method SolidEDTA solution
for Seawater, Milk into
and Solida
1. Calculate the total moles1. of EDTA Eriochrome
Calculate
addedthe the Black
to total moles ofTEDTA indicator solution.
added to the
Samples Samples sample solution.
conical flask. sample solution.
1. Pipette 10 mL of the sample 1. solution
Pipette 10 mLa of
into the sample
conical 2. solution
Calculateintothe
a conical
moles of the2.4. Prepare
Calculate the
magnesium amoles
0.0025
chloride of themol L1 magnesium
magnesium chloride chloride
2. Add 10 mL of ammonia
flask. flask. buffer solution and 1 mL of solution
solution used in the back titrationsolution used in the
from your concordant
by back titration
diluting from
the your concordant 1
0.025 mol L magnesium
2. Eriochrome 0.05 mol L results. From the equation of results. Frombelow,
the titration the equation
the of the titration below, the
Add 20 mL of 0.05Black
mol L2.TEDTA
indicator
Add 20 mL ofsolution.
solution.
1 EDTA solution. 1

moles of Mg will be equivalent2+ moles tochloride

of Mg
the moles willofsolution
be equivalentby
excess 2+ a moles
to the factor of 1/10.
of excess
3. Add 10 mL of ammonia buffer, 3. Add 50 mL10 mL of ammonia buffer, 50 mL of distilled
of distilled
EDTA. EDTA.
3. water
Titrate the EDTA with
and 1 mL of Eriochromewater the
Black and magnesium
1 mL ofsolution.
T indicator chloride
Eriochrome Black T indicator solution. 5. Titrate the+ sample solution with this 0.0025 molL1
4. solution untilwith
Titrate the sample thethe
4.endpoint
Titrate the
standard ismolL
0.025 reached
sample with the EDTA
a permanent
standard -1
4
0.025 molL+ Mg2+ [Mg-EDTA] -1 EDTA2 4
Mg2+ [Mg-EDTA]2
magnesium chloride solutionmagnesium chloride
until a permanent pinksolution until
3. aGivenpermanent
the ratiopink
of Ca + Mg
magnesium
3. :Given EDTA the
2+
= 1 : ratio
chloride solution until a permanent
of Ca + Mg : EDTA = 1 : 1, calculate
1, calculate
2+ 2+ 2+
colour
colour appears.
change fromcolour
blue
appears.
to pink. the moles of Ca and Mg that 2+themust pink
moleshave
2+ofcolour
beenand Mg
Ca appears.
that mustRepeat
2+
have been the titration with further
2+

complexed with EDTA by subtracting complexed the with

excess EDTA
EDTA by subtracting the excess EDTA
4. Having determined
Titration Method for FreshTitration the
or Tap Water average
Method titre
for Fresh or
Samples of the
Tapthe
from Water
total Samples
moles of EDTA fromaddedthe
samples
to total
until concordant
moles of EDTA added to the sample.
the sample.
results (titres agreeing
magnesium chloride
1. Add a 100 mL of the sample solution,
1. solution
Add a 100 mLa of
into determine
250the mLsample the
Thissolution number
result isinto
the amoles
250 mLof Ca This within
andresult
Mg in is the0.1
2+ moles mL)
sample areandobtained.
of Ca
2+ Mg in the sample
2+ 2+
conical flask. used. conical flask.
of moles solution. solution.
2. Prepare a 0.005 mol L EDTA 2. solution
Prepare aby0.005
1 mol L EDTA solution by diluting
diluting 1

5. theGiven
0.05 molthe Mgsolution
L EDTA
1 2+
: EDTA
the
by 0.05 ratio
a factor of
EDTA
molofL 1/10. 1Add:solution
1
1,20calculate
by a factortheof 1/10. Add 20
mL of this diluted EDTA to themL of thissolution.
sample diluted EDTA to the sample solution.
concentration of your EDTA solution.
3. Add 10 mL of the ammonia 3. buffer
Add 10
andmL1 mL
of the
of ammonia buffer and 1 mL of
Eriochrome Black T indicator solution.
Eriochrome Black T indicator solution.

Figure 1 Colour changes for magnesiumFigure 1 Colour

chloride changes in
back-titration forclear
magnesium chloride
Figure 2 back-titration Figure 2chloride
in clear for magnesium
Same colour changes Same colour changes as
back-titration forinmagnesium chloride back-titration as in
Figure 1 Colour
before endpoint changes
(all Ca /Mg
2+
solution
solution using Eriochrome Black T indicator.
for
before
ions complexed
2+
Leftusing
flask:Eriochrome
endpoint
blue colour Black
bymagnesium
excess(all
2+
Ca all
EDTA, /Mg
2+
well T indicator.
chloride
ions
indicator complexed
Left1, flask:
Figure
colourby
but forblue
back-titration
wellexcess
colour
cloudy
EDTA,
before
(opaque)
all indicator
endpoint. Figure
well sampleFigure 1, but
solution,
colour
Centre flask: last trace of2blue/purple
egfor cloudy
milk.
well before
Left (opaque)
Same flask: bluesample solution, eg milk. Left flask: blue
colour
Centre flask:changes
endpoint.colour for magnesium
last trace of blue/purple colour chloride back-
inmolecules
clear solution
uncomplexed). using
Centre Eriochrome
flask: molecules
last trace ofuncomplexed).
beforecomplexed
Black
blue/purple
endpoint (excess
Centre
colour
EDTA
T
just
Mgalmost
indicator.
flask: last
justtrace
totally
before Left
of blue/purple
endpoint. flask:
colour
Right flask: titration
just
justpink/red as
beforeatendpoint.
colour endpoint. in
Right Figure 1,
flask: pink/red but for
colour at cloudy
endpoint. (opaque) sample solution,
2+ complexed by added Mg ).
2+
before endpoint (excess EDTA almost totally by added 2+
).
blue colour
Right flask: pink/redwell before
colour at endpoint
endpointRight
(all flask:complexed
EDTA (all
pink/red colourCaat2+endpoint
by added Mg/Mg ionscomplexed
, (all EDTA
2+ complexed by added Mg ,
2+
eg milk. Left flask: blue colour well before endpoint. Centre
indicator also complexed).
by excess EDTA, all indicator
indicator also complexed).
molecules uncomplexed). Centre flask: last trace of blue/purple. Right flask: pink/red colour at
2flask: last trace of blue/purple 2 colour just before endpoint 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).
 Result Calculations nature and source of the sample it may be necessary to
vary the concentration of the EDTA( if the titre volume is
1. Calculate the total moles of EDTA added to the too low) or to dilute your solutions (if the titre volume
sample solution. is too high). The average titre volume should be in the
2. Calculate the moles of the magnesium chloride range of 10 30 mL.
solution used in the back titration from your
concordant results. From the equation of the titration Contact Us
below, the moles of Mg2+ will be equivalent to the
Safety moles of excess EDTA. If you have
Contact Usany questions or comments relating to this
experiment, please contact us. Please note that this
The concentrated EDTAammonia
4
+ Mg2+solution used in preparing
[Mg-EDTA] 2 If you haveisany
service questions
for senior or comments
school chemistryrelating
studentstointhis
the buffer and indicator solutions is highly corrosive and experiment, please contact us:
New Zealand only. We regret we are unable to respond
3. Given
should be the ratio with
handled of Cacare.
2+
+ Mg 2+
Wear : EDTA
safety =glasses
1 : 1, calculate
and to queries from overseas.
Outreach
the moles
rubber of Ca2+ and Mg2+ that must have been
gloves.
complexed with EDTA by subtracting the excess EDTA College Outreach
of Science
Both the buffer and indicator (and thus also
from the total moles of EDTA added to the sample. the titration College
University of
of Science
Canterbury
solution) will liberate ammonia2+gas to some University
extent. This Private Bag 4800 of Canterbury
This result is the moles of Ca and Mg2+ in the sample
gas may be harmful if inhaled in large quantities. Work Christchurch 4800
Private Bag
solution. Christchurch
in a fumehood or well ventilated area. New Zealand
New Zealand
Phone:
Phone:+64+643 364 2178
3 364 2178
AdditionalNotes
Additional Notes Fax: +64
Fax: +643 3364
3642490
2490
Email:outreach@canterbury.ac.nz
Email: outreach@canterbury.ac.nz
1. 1. Ethylenediaminetetraacetic
Ethylenediaminetetraaceticacid, acid,EDTA
EDTAisisaalarge
large
molecule which creates a complex with a metal ion, www.outreach.canterbury.ac.nz
molecule which creates a complex with a metal ion, www.outreach.canterbury.ac.nz
bondingthrough
bonding throughsixsix coordination
coordination sites.
sites.
2-
O

O O CH2
C CH2
O N
CH2
Ca2+
O N CH2
C CH2
CH2
O O
C

O
Complex formed by EDTA and calcium ions
Complex formed by EDTA and calcium ions
2. The ammonia buffer (pH ~ 10.5) used here is needed
as Eriochrome Black T only changes colour in the pH
2. The7 ammonia
range 11. buffer (pH ~ 10.5) used here is needed
as Eriochrome Black T only changes colour in the pH
3.range
The7 presence
11. of some other metal ions eg
copper, iron, cobalt, nickel, zinc, manganese in
3. The presence of some other metal ions
high concentrations may introduce error to the
eg copper, iron, cobalt, nickel, zinc, manganese
determination of calcium and magnesium ions using
in high concentrations may introduce error to the
this method, although this is unlikely.
determination of calcium and magnesium ions using
4.thisAs the concentration
method, of Ca
although this
2+
and Mg2+ in the sample
is unlikely.
solution may vary considerably2+depending on the nature
4. As the concentration of Ca and Mg2+ in the sample
and source of the sample it may be necessary vary the
solution may vary considerably depending on the
concentration of the EDTA( if the titre volume is too
low) or to dilute your solutions (if the titre volume is too
high). The average titre volume should be in the range of
10 30 mL.