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. 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

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

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

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 Titration Method for Fresh or Tap Water Samples Eriochrome Black T indicator solution. chloride Preparechloride a 0.0025 mol L magnesium Eriochrome Black T indicator solution. magnesium 4. Prepare a 0.0025 mol L 4. of the solution is almost 7 (according to pH indicator magnesium solution by diluting the 0.025 mol L magnesium solution by diluting the 0.025 mol L 3.magnesium Titrate the EDTA with the magnesium chloride 3. Titrate the EDTA with the chloride 1. Add a 100 mL of the sample solution into a 250 mL chloride chloride solution by a factor of 1/10. solution by a factor of 1/10. paper). For eggshells, the inner will remain until themembrane endpoint is reached a permanent solution until the endpoint issolution reached a permanent conical flask. 5. with Titrate sample solution with this 0.0025 molL colour change from blue to pink. 5. Titrate the sample solution thisthe 0.0025 molL colour change from blue may to pink. undissolved and be carefully removed from the magnesium chloride solution until a permanent pink magnesium chloride solution until a permanent pink 4. Having determined the average titre of the 4. Having determined the average titre of the 2. Prepare asamples 0.005 mol with L1 EDTA solution by diluting the solution. Transfer the solution to a 100 mL volumetric colour appears. Repeat the titration further samples appears. Repeat the with further magnesium chloride solution, determine the number of titration magnesium chloride solution, determine the number of colour 1 (titres agreeing within 0.1 mL) until concordant results until concordant results (titres agreeing within 0.1 mL) 0.05 mol L EDTA solution by a factor of 1/10. Add moles used. moles used. flask and make up to the mark with distilled water. are obtained. are obtained. 20mL of this diluted EDTA to the sample solution. : EDTA ratio of 1 : 1, calculate the 5. of Given the Mg the 1 : 1, calculate 5. Given the Mg : EDTA ratio Standardisation of the EDTA concentration ofSolution your EDTA solution. concentration of your EDTA solution. Calculations Result Calculations Result 3. Add 10 mL of the ammonia buffer and 1 mL of Titration Method for Seawater, Milk and Solid 1. Pipette a 10for mL sample of the EDTA solution into a Titration Method Seawater, Milk and Solid 1. of EDTA Calculate the moles ofT EDTA added to the 1. Calculate the total moles added to total the Black Eriochrome indicator solution. Samples Samples sample solution. sample solution. conical flask. 1. solution Pipette 10 mL the sample into a conical 1. Pipette 10 mL of the sample into a of conical 4. Prepare amoles 0.0025 L1 magnesium chloride 2. Calculate the of themol magnesium chloride 2. solution Calculate the moles of the magnesium chloride flask. flask. 2. Add 10 mL of ammonia buffer solution and 1 mL of solution used in the back titration from your concordant 1 solution used in the back titrationsolution from your concordant by diluting the 0.025 mol L magnesium Frombelow, the equation From the equation of results. the titration the of the titration below, the EDTA solution. Add 20 mL ofsolution. 0.05 mol L results. EDTA solution. 2. Eriochrome Add 20 mL of 0.05 mol L2. Black T indicator chloride a moles factor of 1/10. will be equivalentby to the of excess moles of Mg to the moles ofsolution excess moles of Mg will be equivalent 3. Add 10 mL of ammonia buffer, 50 mL of distilled 3. Add 10 mL of ammonia buffer, 50 mL of distilled EDTA. EDTA. 3. Titrate the EDTA with the magnesium chloride 1 mL ofsolution. Eriochrome Black T indicator solution. water and 1 mL of Eriochromewater Black and T indicator 5. Titrate the sample solution with this 0.0025 molL1 4 2 4 EDTA + Mg2+ [Mg-EDTA]2 EDTA + Mg2+ [Mg-EDTA] 4. Titrate the sample with the standard 0.025 molL untilwith the endpoint ismolL reached a permanent 4. solution Titrate the sample the standard 0.025 magnesium chloride solution until a permanent chloride solution until permanent pink 3. :Given of Ca + Mg : EDTA = 1 : 1, calculate magnesium chloride solutionmagnesium until a permanent pink 3. a Given the ratio of Ca + Mg EDTA the = 1 : ratio 1, calculate colour change from blue to pink. colour appears. and Mg that mustRepeat have been the titration with further the moles ofcolour Ca pink appears. colour appears. must have been the moles of Ca and Mg that complexed EDTA by subtracting the excess EDTA complexed with EDTA by subtracting the with excess EDTA 4. Having determined the average titre of the samples until concordant results (titres agreeing Method for Fresh or Tap Water Titration Method for FreshTitration or Tap Water Samples the moles of EDTA added to the sample. from the total Samples moles of EDTA from added to total the sample. magnesium chloride solution, determine the number within 0.1 mL) are obtained. 1. solution Add a 100 mL the sample solution into 250 mL 1. Add a 100 mL of the sample into a of 250 mL and Mg in the sample is the moles of Ca andresult Mg in sample This result is the a moles of Ca This conical flask. used. conical flask. solution. solution. of moles 2. solution Prepare a 0.005 mol L EDTA solution by diluting by diluting 2. Prepare a 0.005 mol L EDTA 2+ EDTA by a factor of 1/10. Add 20 the mol L 1/10. 5. Given Mg : EDTA ratio of 1 :solution 1, calculate the solution by 0.05 a factor of Add 20 the 0.05 molthe L EDTA mL of this diluted EDTA to the sample solution. mL of this diluted EDTA to the sample solution. concentration of your EDTA solution.
1 1 1 1 1 1 2+ 2+ 1 1 2+ 2+ -1 -1 2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+ 1 1 1 1

Titration Method for Seawater, Milk and Solid Samples 1. Pipette 10 mL of the sample solution into a conical flask. 2. Add 20 mL of 0.05 mol L1 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 molL1 magnesium chloride solution until a permanent pink colour appears.

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

Figure 1, but cloudy sample solution, eg milk. Left flask: blue solution Left blue colour well sample Figure 1, flask: but for cloudy (opaque) solution, egfor milk. Left (opaque) flask: blue solution using Eriochrome Black T indicator. Leftusing flask:Eriochrome blue colour Black well T indicator. Figure 1 Colour changes for magnesium chloride back-titration Figure 2 Same colour changes for magnesium chloride backcolour well before endpoint. Centre flask: last trace of blue/purple colour before endpoint Ca all /Mg ions complexed excess EDTA, all indicator colourby well before endpoint. Centre flask: last trace of blue/purple colour before endpoint (all Ca /Mg ions complexed by excess(all EDTA, indicator just before endpoint. Right flask: pink/red colour at endpoint. uncomplexed). Centre flask: last trace of blue/purple colour just just before endpoint. Right flask: pink/red colour at endpoint. molecules uncomplexed). Centre flask: molecules last trace of blue/purple colour just in clear solution using Eriochrome Black T indicator. Left flask: titration as in Figure 1, but for cloudy (opaque) sample solution, before endpoint (excess EDTA almost totally before endpoint (excess EDTA almost totally complexed by added Mg2+ ). 2+ complexed by added Mg ). blue colour well before endpoint (all Ca /Mg ions complexed eg milk. Left flask: blue colour well before endpoint. Centre flask: pink/red colour at endpoint complexed by added Mg , Right flask: pink/red colour at endpointRight (all EDTA complexed by added Mg , (all EDTA indicator also complexed). indicator also complexed). by excess EDTA, all indicator molecules uncomplexed). Centre flask: last trace of blue/purple. Right flask: pink/red colour at 2 2 flask: last trace of blue/purple 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).
2+ 2+ 2+ 2+ 2+ 2+ 2+ 2+

Figure 2 Same colour changes as forin magnesium chloride back-titration as in Figure 1 Colour changes in for magnesium chloride in clear for magnesium Figure 2 back-titration Same colour changes chloride back-titration Figure 1 Colour changes for magnesium chloride back-titration clear

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 Contact Us below, the moles of Mg2+ will be equivalent to the If you have any questions or comments relating to this Safety Contact Us moles of excess EDTA. experiment, please contact us. Please note that this 4 2 The concentrated ammonia solution used in preparing If you have questions or comments toin this service isany for senior school chemistryrelating students EDTA + Mg2+ [Mg-EDTA] the buffer and indicator solutions is highly corrosive and experiment, please contact us: New Zealand only. We regret we are unable to respond 2+ 2+ 3. Given ratio with of Ca + Mg : EDTA =glasses 1 : 1, calculate should be the handled care. Wear safety and to queries from overseas. Outreach the moles of Ca2+ and Mg2+ that must have been rubber gloves. Outreach of Science complexed with EDTA by subtracting the excess EDTA College College of University of Science Canterbury Both the buffer and indicator (and thus also the titration from the total moles of EDTA added to the sample. University of Canterbury solution) will liberate ammonia2+gas to some extent. This Private Bag 4800 This result is the moles of Ca and Mg2+ in the sample Private Bag gas may be harmful if inhaled in large quantities. Work Christchurch 4800 solution. Christchurch in a fumehood or well ventilated area. New Zealand New Zealand Phone: +64 3 364 2178 Phone: +64 3 364 2178 Additional Notes Fax: +64 364 2490 Fax: +64 33 364 2490 Additional Notes Email:outreach@canterbury.ac.nz outreach@canterbury.ac.nz Email: acid, EDTA is aa large 1. 1. Ethylenediaminetetraacetic Ethylenediaminetetraacetic acid, EDTA is large www.outreach.canterbury.ac.nz molecule which creates a complex with a metal ion, www.outreach.canterbury.ac.nz molecule which creates a complex with a metal ion, bonding through six coordination sites. bonding through six coordination sites.
2-

nature and source of the sample it may be necessary to vary 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.

C
O

O
C
CH2

CH2
N
N

Ca2+
C

CH2
CH2

CH2 O C

CH2

O
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. The ammonia buffer (pH ~ 10.5) used here is needed range 7 11. as Eriochrome Black T only changes colour in the pH 3.range The of some other metal ions eg 7 presence 11. 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 2+ and Mg2+ in the sample 4.thisAs the concentration of Ca method, although this 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.

Complex formed by EDTA and calcium ions