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University of Malta Department of Chemistry CHE1 700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 1:

MELTING POINT DETERMINATION

Introduction

The determination of melting points in capillary tubes is a very simple, effective and rapid way to confirm the identity and purity of organic solid substances.

The process of ‘melting’ is usually characterized by:

(a)

Pronounced softening or shrinking.

(b)

Most of the substance suddenly running down and forming a meniscus.

(c)

All the substance becomes liquid.

Although in some cases (a)-(c) occur within a range of 1 o C, melting usually occurs (especially impure samples) over a temperature range of c. 5-10 o C. In such cases, this range should be recorded (rather than estimating an 'average' reading). Note that even in such case we still refer to the ‘range’ as the melting ‘point’.

Extensive databases now exist listing melting points of pure substances including:

o

The data collection of the National Institute of Standards and Technology (NIST) which is available online on http://webbook.nist.gov/;

o

The Organic Compounds Database maintained by Colby College available online on http://www.colby.edu/chemistry/cmp/cmp.html;

o

The Sigma-Aldrich catalogue available online on http://www.sigma-aldrich.com/

o

ChemFinder

from

Cambridge

Software,

available

online

from

 

o

The CRC Chemistry and Physics databook (available in the library)

 

o The Materials Safety Data Sheets (MSDS, may be accessed from manufacturers websites, see website for details.). Published melting points are usually within a 0 - 360 o C temperature range and refer to

values taken in capillary tubes sealed at one end * .

It is interesting to note that melting point values for the same material quoted from different data sources may be slightly different. One should also bear in mind that the melting point ranges listed in the company catalogues (e.g. on the Sigma-Aldrich website) are the melting points of the compounds as they are sold, i.e. if the compounds are sold slightly impure, then the company catalogue melting point will deviate from those listed in the CRC data book or on the MSDS which always refer to the melting point of the pure compound.

* Your laboratory demonstartor should show you how to prepare these capillary tubes sealed from one end from a normal capillary tube.

The recommended procedure for melting point determination is to first place a small amount of the sample on a clean dry watch-glass or glass slide and then powder it by means of a spatula or the rounded end of a glass rod. You should then fill four capillary tubes each to a depth of 2-4 millimetres with the powdered sample. (This is best performed by tapping the open end of the capillary tube into the powdered sample and then tapping the sealed end on the bench to move the powder down the capillary tube. This filling/tapping may be repeated if necessary until you obtain a loose but continuous column of c. 3mm in depth at the sealed end of the capillary tube.) One of these tubes is then placed in the melting point apparatus and the temperature is raised at a relatively fast rate (about 10 °C/minute) until the compound first begins to melt. This temperature is probably an over-estimation of the true melting point and should only be treated as a rough estimate. The melting point apparatus should then be allowed to cool to about 20 °C below that estimated temperature, and the process is repeated with another sample, this time using a much slower heating rate (less than 1 o C per 30 seconds for the last 3 or 4 degrees). This reading should provide you with the first accurate melting point result. At least one other accurate reading should then taken using the third sample. If these two accurate readings are inconsistent, then a third, and possibly more determinations have to be performed until the results become consistent. The melting point should then be taken as the average range. For example, if the three accurate readings were made, and these are a 1 -b 1 , a 2 -b 2 and a 3 -b 3 then the average range is from (a 1 +a 2 +a 3 )/3 to (b 1 +b 2 +b 3 )/3 .

NOTE: You should always discard melting point tubes once they have been used. If the melting point obtained is doubtful repeat using a fresh tube.

Procedure:

NOTE: Before you commence the practical work, please ‘read’ carefully the Safety Data for A –E and M1 provided in the table below.

You have been provided with six samples A – E and M1, the molecular formula and melting points for some of which are given below.

Sample

Molecular

Melting pt. ( o C) *

Risks / Safety data

Formula

A

C

7 H 7 NO 2

54

Toxic, R23/24/25, R36/37/38, S45, S26, S22, S36/37/39

B

C

7 H 6 O 2

 

Harmful, R22, R41, R37/38, R42/43, S26, S36, S22

C

C

7 H 6 O 3

159

Toxic, R61, R22, R41, R37/38, S45, S26, S36/37/39, S22

D

C

7 H 5 NO 4

239

Harmful, R22, R41, S26, S39

E

C

9 H 8 O 2

131-134

Irritant, R36/37/38, S26, S36

M1

not available

 

similar to one / a combination of A-E

(*) Melting points quoted directly from the information supplied by manufacturer of the respective chemicals on the labels of the original containers.

Please refer to the RISK and SAFETY terms and symbols on the course website,

http://staff.um.edu.mt/jgri1/teaching/che1302

You should first determine the meting point of substances B and M1 individually using the procedure outlined above. Enter these results in your laboratory notebook. (After the practical, you should the search the online Organic Compounds Database maintained by Colby College to try and identify substances A-E. Report the name and structural formula of these compounds in your lab reports. You may also wish to consult other databases to check whether there are any differences in the reported values, etc.)

You should find that substance M1 has a melting point that is very similar to one of the other five samples. This similarity in melting points may indicate that substance M1 is of the same chemical composition as one of the samples A-E (henceforth referred to M2).

You can test this hypothesis by (1) Preparing a ‘new’ sample (=M3) by grinding mixture of equal amounts of M1 and M2, (2) Filling three separate tubes with M1, M2 and M3 respectively, and (3) placing them together in the melting point apparatus with the mixture in the middle.

If M1 and M2 are identical, then M1, M2 and M3 should all melt simultaneously. However, if this is not the case, M3 will melt over a range of temperature lower than M1 and M2. In such case, you should determine the melting point of M3 more accurately using the procedure outlined above and enter the value in your laboratory notebook.

Once you have convinced yourself that your results are consistent you should fill in the R-CHE1700-01 form attached (University Copy) and hand it to the laboratory demonstrator. The demonstrator should also check your laboratory notebook and fill in the Chemistry Practical Front Matter sheet. Note that this procedure should be repeated for every practical.

University of Malta Department of Chemistry CHE1 700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 2:

RECRYSTALLISATION AS A TECHNIQUE FOR PURIFICATION

Introduction

The purification of organic compounds is one of the commonest procedures encountered in experimental organic chemistry. This is because it is very rare that a product from a chemical reaction can be extracted in a pure form. The common practice to purify a product which is crystalline in its pure form is through a process of one / a series of crystallisation / recrystallisation process/es,

In its simplest form, the recommended procedure for recrystallisation is as follows * :

1. An inert solvent is chosen such that the substance to be crystallised is only slightly soluble in cold solvent but very soluble in hot solvent. (This is discussed at a later stage.)

2. The impure substance to be re-crystallized is dissolved in the minimum amount of hot solvent (at or near its boiling point). Note: (i) Please refer to page 3 for a

discussion on how to choose a solvent. (ii) The only case when it is justified (and recommended) to use more solvent than the minimum amount is when there is a very steep solubility curve, i.e. when slight cooling causes considerable crystallisation.

3. The hot solution is filtered rapidly (to remove any insoluble particles e.g. dust) using a stemless funnel and a fluted filter paper (see Fig. 1) into a conical flask. Note that both the funnel and the flask should be warmed just a little before filtration by passing them over a flame so as to prevent premature crystallisation. (If this occurs, the crystals have to be re-dissolved.)

4. The conical flask containing the hot filtered solution is then corked lightly and then left undisturbed to cool down very slowly hence allowing the dissolved substance to crystallise out. Note that slow cooling favours formation of large crystals, as opposed to quenching which will result in the formation of a powder. If an ‘oil’ emerges rather than crystals, one should induce crystallisation by scratching the inside of the conical flask with a glass rod. If this is unsuccessful, one may (a) inoculate (seed) the solution with some of the solid materials, or, (b) cool the solution to lower temperature (e.g. by immersing in an acetone/dry ice bath.

* This procedure has to be slighly modified if the amount of sample to be purified is less than 1g and expensive to obtain (e.g. the final product of a multi-step reaction). In such cases, the filtartion in step (2) should be avoided unless absolutely necessary whilst the filtration in step (5) is carried out using a Willstatter ‘filtration nail’. For further details on such techniques, or on other modifications in special cases, please refer to specialist textbooks, such as Vogel’s Textbook of Practical Organic Chemistry, 5 th Edn. (Longman, 1988).

5.

Once the hot solution has cooled down to room temperature, the conical flask with the solution is immersed in an ice bath to maximise the extraction of the substance from the supernatant solution.

6. The crystals are then separated from the supernatant solution by filtration. This filtration is usually carried out under suction (see fig. 2a) using through a Buchner funnel fitted with a filter paper wetted with the solvent (for large quantities of crystals) or a Hirsch funnel (for medium quantities see Fig. 2b).

7. The crystals are then washed from any remaining impure supernatant solution using minimum amount of ice-cold pure solvent (taking care that the crystals do not dissolve). To do this one must (i) discontinue the suction, (ii) pour small amounts of the chilled solvent, (iii) stir cautiously the solvent/crystals to ensure that the solvent washes all the crystals, and (iv) re-apply suction to ‘dry’ the crystals. Note that this ‘drying’ may be aided by pressing the crystals on the sieve using a clean spatula/flattened glass rod, and by leaving the ‘suction process’ running for an extra few minutes.

8. The solid crystals are then transferred to a watch glass and dried to constant weight in a desiccator under vacuum. Other drying methods include the use of suitably heated ovens or even by placing on a water bath if the substance melts over 100 o C (although this technique may result in contamination of the crystals by steam).

9. The DRY crystals are tested for purity (e.g. by a melting point verification, or by thin layer chromatography). If the substance is still found to be impure, then the whole process is repeated from step 2 until purity can be ascertained or successive recrystallisations result in very little change in the purity (indicated for example by consistency in the melting point values).

for example by consistency in the melting point values). Fig. 1: To prepare a fluted filter

Fig. 1: To prepare a fluted filter paper you must first fold a normal filter paper in half and again in quarters, and then open it up as shown in (a). The edge 2,1 is then folded on to 2,4 and edge 2,3 on to 2,4, hence producing new folds at 2,5 and 2,6 when the paper is opened. The folding is continued, 2,1 to 2,6 and 2,3 to 2,5, thus producing folds at 2,7 and 2,8 respectively (b); further 2,3 to 2,6 giving 2,9, and 2,1 to 2,5 giving 2,10 (c). The final operation consists in making a fold in each of the eight segments - between 2,3 and 2,9, between 2,9 and 2,6, etc. - in a direction opposite to the first series of folds, i.e. the folds are made outwards rather than inwards as first. The result is a fan arrangement (d), and upon opening, the finished fluted filter paper. (e). (Taken from Vogel’s Textbook of Practical Organic Chemistry, 5 th edn.).

Fig. 2: (a) The set-up to be used for filtration under suction using a Buchner

Fig. 2: (a) The set-up to be used for filtration under suction using a Buchner funnel, (b) An illustration of a Hirsh funnel which should be used for crystal quantities in the range of 1-2g. Note that the smaller versions of Hirsch funnels will accommodate filter papers 3-4mm in diameter. Note also that apart from the porcelain versions of the Buchner and Hirsch funnels where the diameter of the sieve pores are in the millimetre region, one may also find sintered glass funnels available in a number of porosities (coarse, medium and fine).

The success behind purification by recrystallisation relies on the fact that the impurities are present in much lower quantities that the substance to be purified, and not necessarily that the impurities are more soluble than the substance to be purified. To illustrate this we present here idealised calculations for two particular (simplified) examples of recrystallisation where we want to extract A from two 10g mixtures of A/B containing (w/w):

(1) 95% A and 5% B (the impurity), see Table 1, and, (2) 90% A and 10% B (the impurity), see Table 2. For each of these two examples we shall consider two scenarios:

(i)

When A is more soluble in cold solvent than the impurity B, and,

(ii)

When the impurity B is more soluble in cold solvent than A.

In the discussion, we shall assume that both A and B are freely soluble in hot solvent and

that 100mL of solvent is used in each recrystallisation.

The calculations presented in Tables 1 and 2 illustrate two important ideas:

1. In all cases, it is possible to obtain A in a pure form, even when irrespective on whether A is more soluble than B in cold solvent, or not.

2. The small change of a 5% increase in the percentage of the impurity B will result in a substantial increase in (a) effort required to obtain 100% pure A in terms of the number of recrystallisations required, and, (b) loss of percentage recovery of A (and we are assuming that there are no losses due to experimental errors/limitations!).

Scenario:

(i)

 

(ii)

 

Substance:

A

 

B

A

 

B

Solubility in cold solvent (g / 100mL)

0.6

0.3

0.3

0.6

1 st Recrystallisation:

       

Initial percentage composition (%):

95

5

95

5

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition: (%)

9.5

0.5

9.5

0.5

0.6

0.3

0.3

0.5

(all)

8.9

0.2

9.2

0

97.2

2.8

100

0

2 ns Recrystallisation:

       

Initial percentage composition (%):

97.2

2.8

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition (%):

8.9

0.2

not

not

0.6

0.2

(all)

required

required

8.3

0

100

0

Total % of A recovered:

87.4%

 

96.8%

 

Table 1: Example 1 - Initial composition is 95%A, 5%B

Scenario:

(i)

 

(ii)

 

Substance:

A

 

B

A

 

B

Solubility in cold solvent (g / 100ml)

0.6

0.3

0.3

0.6

1 st Recrystallisation:

       

Initial percentage composition (%):

90

10

90

10

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition: (%)

9.0

1.0

9.0

1.0

0.6

0.3

0.3

0.6

8.4

0.7

8.7

0.4

92.3

7.7

95.6

4.4

2 ns Recrystallisation:

       

Initial percentage composition (%):

92.3

7.7

95.6

4.4

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition (%):

8.4

0.7

8.9

0.4

0.6

0.3

0.6

0.4

(all)

7.8

0.4

8.3

0

95.1

4.9

100

0

3 rd Recrystallisation:

       

Initial percentage composition (%):

95.1

4.9

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition (%):

7.8

0.4

not

not

0.6

0.3

required

required

7.2

0.1

98.4

1.6

4 th Recrystallisation:

       

Initial percentage composition (%):

98.4

1.6

Initial amount present (g) Amount dissolved in solution at room temperature (g) Amount recovered (g) Final percentage composition (%):

7.2

0.1

not

not

0.6

0.1

(all)

required

required

6.6

0

100

0

Total % of A recovered:

73.3%

 

92.2%

 

Table 2: Example 2 - Initial composition is 90%A, 10%B

Choice of Solvent

The most desirable characteristics of a solvent for recrystallisation are as follows:

1. The solvent must be inert, i.e. it must not react chemically with the substance to be

purified;

2. The substance to be purified must be highly soluble in the solvent at elevated temperatures and ‘insoluble’ at room temperature;

3. The solvent should dissolve the impurities readily or to only a very small extent (so that the impurity may be removed in the first filtration using the fluted filter paper);

4. The solvent should allow the formation of well-formed crystals of the purified compound;

5. The solvent must have a relatively low boiling point so that it may be easily removed from the crystals of the purified compound;

6. Ideally, the solvent should not be toxic, flammable and expensive.

Some common solvents used in recrystallisation are listed in Table 3 below (broadly in the order of decreasing polarity).

Solvent

b.p. (°C)

Comments:

Water (distilled)

100

To be used whenever suitable

Methanol*

64.5

Flammable, toxic

Ethanol

78

Flammable

Industrial spirit

77-82

Flammable

Rectified spirit

78

Flammable

Acetone

56

Flammable

Ethyl acetate

78

Flammable

Acetic acid (glacial)

118

Not very flammable, pungent vapours

Dichloromethane (methylene chloride)*

41

Non-flammable; toxic

Chloroform*

61

Non-flammable; vapour toxic

Diethyl ether

35

Flammable, avoid whenever possible

Benzene*+

80

Flammable, vapour highly toxic

Toluene*+

110

Flammable, vapour toxic

Carbon tetrachloride*

77

Non-flammable, vapour toxic

Light petroleum (petroleum ether, see note)

40-60**

Flammable

Cyclohexane

81

Flammable

Table 3: Common solvents for recrystallisation (Taken from from Vogel’s Textbook of Practical Organic Chemistry, 5 th edn.). Note: (*) To be used in a fume hood, (+) Benzene is now known to be extremely toxic. Use toluene instead whenever possible. (**) See note on petroleum ether on page 6.

It should be noted that:

The use of ether (diethyl ether) as a solvent for recrystallisation should be avoided wherever possible, partly owing to its great flammability (and its potential to form explosive peroxides with prolonged exposure to air, an important factor to consider when handling ‘aged’ ether) and partly owing to its tendency to creep up

walls of the containing vessel, thus depositing solid matter by complete evaporation instead of preferential crystallisation.

The vapours of methanol, dichloromethane, benzene, toluene and carbon tetrachloride are toxic and therefore recrystallisations involving their use must he conducted in an efficient fume cupboard: excessive inhalation of any vapour should be avoided.

Toluene is much less toxic than benzene and should be used in place of benzene whenever possible.

Note on petroleum ether: Petroleum ether (or light ether) is a mixture of alkanes and is available in different compositions labelled according to the boiling points. Available fractions include b.p. 40-60, 60-80, 80-100 and 100-200 o C. When the boiling point exceeds 120 o C the fraction is usually called 'ligroin'. Pure pentane (b.p. 36 o C), hexane (b.p. 69 o C), and heptane (b.p. 98 o C), are also frequently used as recrystallisation solvents, although these are much more expensive than the mixtures.

There are no hard and fast rules on the choice of solvents. In general, it should be noted that a substance is likely to be most soluble in a solvent to which it is most closely related in chemical and physical characteristics. In particular, a polar substance is more soluble in polar solvents and less soluble in non-polar solvents (“like dissolves like”). It is also common practice to use a mixture of two or more solvents when one cannot find a single solvent with the right characteristics.

In practice the choice of a solvent for recrystallisation must be determined experimentally if no information is already available. The recommended procedure to do this is as follows:

1. About 0.lg of the powdered substance is placed in a small test tube

2. The cold solvent is added a drop at a time with continuous shaking of the test tube. If the sample dissolves easily in 1 mL of cold solvent the solvent is unsuitable.

3. If after about 1 mL of the solvent has been added, the substance is still undissolved, then the mixture is heated to boiling to check the solubility in hot solvent. If the sample dissolves easily upon gentle warming, the solvent is un- suitable.

4. If all the solid does not dissolve, more solvent is added in 0.5mL portions, and again heated to boiling after each addition. If 3 mL of solvent is added and the substance does not dissolve on heating, the substance is regarded as sparingly soluble in that solvent, and hence unsuitable.

5. If the compound dissolves in the hot solvent, the tube is cooled to determine whether crystallisation occurs. (Note: If crystallisation does not take place rapidly, you should check if this is due to the absence of suitable nuclei sites by scratching the tube below the surface of the solution with a glass rod. The fine scratches on the walls and the minute fragments of glass produced may serve as excellent nuclei for crystal growth.) If crystals do not separate (even after scratching for several minutes and cooling in an ice-salt mixture), the solvent is considered as unsuitable.

6. If crystals separate, the amount of these should be noted.

7. This process may be repeated with other solvents (using a fresh test tube for each experiment) until the best solvent is found. For each experiment, the approximate proportions of the solute and solvent giving the most satisfactory results should be

recorded. Note that if no solvent is found to be satisfactory, one should attempt using mixed solvents (see note below).

Note on the use of mixed solvents:

If the substance is found to be far too soluble in one solvent and much too insoluble in another solvent to allow recrystallisation, mixed solvents or solvent pairs may be used. The two solvents must, of course, be completely miscible. (Miscible solvents must not have very different polarities.) The procedure for use in such cases is as follows:

o

The compound is dissolved in the solvent in which it is very soluble.

o

The hot solvent, in which the substance is only sparingly soluble, is added cautiously until a slight turbidity is produced.

o The turbidity is then just cleared by the addition of a small quantity of the first solvent and the mixture is allowed to cool to room temperature. This should result in crystal formation. Pairs of liquids which may be used include: alcohols and water; alcohols and toluene; toluene and light petroleum; acetone and light petroleum; diethyl ether and pentane; glacial acetic acid and water; dimethylformamide with either water or toluene.

Procedure

In this practical you shall first check the suitability of the following eight solvents

o

water

o

ethanol

o

petroleum ether 60-80 o C

o

hexane

o

dichloromethane

o

toluene

o

acetone

o

ethyl acetate

for recrystallisation of:

(A)

Benzoic acid

(B)

Acetanilide, and

(C)

Naphthalene

and then purify benzoic acid (m.pt. 122 o C) from a mixture of benzoic acid and 5% naphthalene (w/w).

In particular:

(a)

Before the lab session, you should look up:

I.

The safety information for all the chemicals to be used from the MSDS datasheets and fill in the special assessment forms where appropriate.

II. The boiling points of the solvents and the meting points of A-C.

(b)

Test the suitability of the different solvents for recrystallisation of A, B and C by

following a modified (because of time constraints) version of the procedure on page 6, i.e.:

i. Place about 0.2g of the powdered substance in a small test tube;

ii. Add 1mL of cold solvent & check for solubility;

iii. Bring to the boil using the water baths provided and check for solubility in hot solvent;

iv. Allow the mixtures to cool (slowly) and check for the formation of

(c)

crystals.

Having identified the best solvent to use for the purification of benzoic acid though recrystallisation from a mixture of benzoic acid and 5% naphthalene (w/w), you should place about 1g (weighed) into a flat-bottomed flask and carry out one recrystallisation procedure as outlined on pages 1-2. The only modification (because of time) is that you should dry your crystals (step 9) first between two filter papers, and then in a flameless oven set at a temperature in

between the boiling point of the solvent and the melting point of the benzoic acid. Having dried the solvent you should:

i. Measure the percentage extraction of benzoic acid.

ii. Check the purity of your crystals through a melting point determination. (Recall that pure benzoic acid melts at 122 o C.)

Because all (apart from water and dichloromethane) of the solvents are flammable, you should not heat the solvents on a naked flame. Note that all the organic solvents you shall be using apart from toluene (b.pt. 110 o C) have boiling points less than 100 o C.

University of Malta Department of Chemistry CHE1 700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 3: INVESTIGATING THE BROMINATION OF

ACETANILIDE – An Example of an Electrophilic Aromatic Substitution

Introduction

Bromine reacts with the aromatic amide acetanilide (N-phenylethanamide, 1) in a substitution mode in which one or more hydrogen atoms bonded to the benzene ring are replaced by bromine atoms.

CH 3 H N O
CH 3
H
N
O

(1)

The reaction can be described as follows:

CH CONHC H

3

65

+ n

Br

2

CH CONHC H

3

65- n

Br

n

+ n

HBr

Although at a first glance it may appear that a large number of products can result from this reaction, in practice only one major product forms when this reaction is carried out at room temperature * . For example, meta monosubstituted products can be excluded since the–NHCOCH 3 group is ortho / para directing. Table 1 lists the melting points of the possible products. It may be observed that these different potential products have very different melting points, and hence, the identity of the reaction product should be determined without difficulty even if the crude compound is used in the melting point determination .

2-Bromoacetanilide

99

4-Bromoacetanilide

165.4

2,4-Dibromoacetanilide

145.4

2,6-Dibromoacetanilide

208-209

2,4,6-Tribromoacetanilide

232

Table 1: The Melting Points ( o C) of possible bromination products (from Beilstein).

* The Risks/Safety phrases for this product (taken from the Fisher Scientific (Canada) website, http://www.fishersci.ca/ ) are listed in Appendix 1. The presence of impurities, (e.g. any unreacted acetanilide, or other minor reaction products) will result in a lowering of the melting point. Thus, unless properly purified, the product of bromination will most probably not melt at the exact temperature listed in the table.

Experimental Procedure

(a)

Before your practical, you should look up the safety information for acetanilide, glacial acetic acid, bromine liquid (used to prepare bromine solution) and ethanol from the MSDS datasheets and fill in the special assessment forms where appropriate (see http://staff.um.edu.mt/jgri1/safety/download ).

(b)

Carry out the reaction using the following procedure:

i. Place 0.5g of acetanilide (accurately weighed) and 5mL of glacial acetic acid in a boiling tube which is secured using a clamp and placed in a beaker containing water at room temperature.

ii. Stir carefully but effectively the acetanilide/glacial acetic acid mixture until all the solid acetanilide dissolves.

iii. Add with stirring, 6mL bromine solution and continue stirring for another 20 minutes.

iv. At the end of this period, quench the reaction by adding 30mL of water and 6mL of saturated sodium hydrogen sulfite solution, again keeping the mixture well stirred. Continue stirring until the red colour (indicating an excess of the bromine) disappears.

v. Cool the tube in an ice-water bath for 15 minutes.

vi. Collect the product by vacuum filtration, wash with cold distilled water and

air-dry. Determine the weight of crude product, and hence determine the percentage yield.

(c)

Product characterization: Determine the melting point of the product and hence identify it using the data provided in Table 1.

(d)

If you have time, recrystallise your product from ethanol and repeat the melting point determination.

(e)

In your discussion, use the information given in Appendix 2, and other information you may regard are important to explain the outcome of this experiment.

Appendix 1:

The Risks/Safety information for the main product of this reaction (taken from the Fisher Scientific (Canada) website, http://www.fishersci.ca/

European Labeling in Accordance with EC Directives Hazard Symbols: Not available. Risk Phrases:

Safety Phrases:

S 24/25 Avoid contact with skin and eyes.

S 28A After contact with skin, wash immediately with plenty of water.

S

37 Wear suitable gloves.

S

45

In case of accident or if you feel unwell, seek

medical advice immediately (show the label where

possible).

Appendix 2: Orientation of Nitration in Substituted Benzenes (at 25 o C)

X X HNO 3 / H 2 SO 4
X
X
HNO 3 / H 2 SO 4

Meta-directing deactivators

NO 2

 

X

 

% Ortho

% Meta

% Para

+

2

89

11

N CH

(

3

)

3

NO

2

7

91

2

CN

 

22

77

2

COOH

 

17

81

2

COOCH

2

CH

3

28

66

6

COCH

3

26

72

2

CHO

 

19

72

9

Taken from Jojn McMurry, ‘Organic Chemistry’, 3rd ed., Brooks/Cole Publsihing Company, USA (1992), pg. 579.

Ortho/para-directing deactivators

X

% Ortho

% Meta

% Para

F

13

1

86

Cl

735

1

64

Br

43

1

56

I

45

1

54

Ortho/para-directing activators

X

% Ortho

% Meta

% Para

CH

3

63

3

34

OH

50

0

50

NHCOCH

3

19

2

79

University of Malta Department of Chemistry CHE 1700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 4: SEPARATION BY EXTRACTION

Introduction

Separation by extraction is a technique which allows the separation of compounds based on their difference in solubility in two immiscible solvents.

If a solute X is allowed to distribute itself between two immiscible solvents, A and B, then the following equilibrium will be established:

XA()

!!"

XB()

#!! The equilibrium constant for this process is known as the distribution coefficient or the partition coefficient, K D , or B D A and is given by:

K

D

()

X

==D

B

A

()

X

the concentration of X in phase B

=

the concentration of X in phase A

[

X

]

B

[ X

] A

Similarly, for solutes X and Y distributed into two immiscible solvents, A and B, we may define the following equilibria:

XA()

YA()

#!! !!"

!!"

XB()

YB()

#!! and for effective separation by extraction of X and Y using solvents A and B, we require that:

K

D

()X

>> K

D

()Y

in which case most of X will find itself in solvent B and most of Y will find itself in solvent A, or,

K

D

()X

<< K

D

()Y

in which case most of X will find itself in solvent A and most of Y will find itself in solvent B.

Solvents A and B are usually water (the aqueous phase) and a water-immiscible organic solvent, such as diethyl ether respectively (henceforth referred to as ether), di-isopropyl ether, toluene, dichloromethane and light petroleum.

The selection of the organic solvent will depend upon the solubility of the substance to be extracted in that solvent and upon the ease with which the solvent can be separated from the solute. Diethyl ether is very frequently used since:

1. It is very immiscible with water;

2. It has very powerful solvent properties for non-polar solutes;

3. It has a very low boiling point (35 °C) hence it can be evaporated easily after the extraction

However, special care must be taken when handling this solvent since it is extremely volatile and flammable * . In particular, all naked flames in the immediate vicinity should be extinguished.

Extractions are carried out using a separatory funnel (conical, as in Fig. 1, or pear-shaped) which should have a short stem and fitted with a ground glass interchangeable stopper. The size of the separatory funnel must be about twice the volume to be extracted. It is usually convenient to have this separatory funnel mounted in a ring on a stand with a firm base.

It is also important to check that the stopcock of the funnel is not jammed. For best performance, the barrel and plug of the stopcock are dried with a linen cloth and in the case of glass stopcocks lightly treated with a suitable lubricant such as silicon grease or petroleum jelly.

lubricant such as silicon grease or petroleum jelly. Fig. 1: A conical separating funnel The solution

Fig. 1: A conical separating funnel

The solution and the extraction solvent (in this case exemplified by ether/water) are introduced into the funnel, and with the stopper firmly held in place the funnel is shaken gently so as to avoid a quick build-up of the internal pressure. Furthermore, it is essential to periodically invert the funnel and to open the stopcock to relieve the excess pressure. This gentle shaking / relieving of pressure should be continued until the atmosphere in- side the funnel is saturated with ether vapour. At this stage, further shaking develops little or no additional pressure. The separating funnel is then shaken vigorously for 2-3 minutes to ensure the maximum possible transfer of the solutes to the appropriate phase and

* The fire hazard is reduced also by employing di-isopropyl ether (b.p. 67.5 °C), but this solvent is much more expensive than diethyl ether.

returned to the stand to allow the mixture to settle. When two sharply defined layers have formed, the lower aqueous layer is run off and separated as completely as possible. However, care must be taken not to run off drops of the ethereal phase. In fact, it would be appropriate to run down the last few drops of the aqueous phase and the first few drops of the ethereal phase into a small beaker so as to avoid mutual contamination of the two phases.

The separated solutes are then removed from solution. This can either be done by evaporation of the solvent (especially for the organic phases where the solvents are usually very volatile), or by precipitation. The solutes are then dried, purified by recrystallisation, and tested for purity using a melting point determination or a chromatographic method.

Procedure

For this practical, you have been provided with a mixture containing benzoic acid (50% by mass) and 4-nitrotoluene. You are required to:

!"Isolate these two components through extractions; !"Purify the separated samples through recrystallisations; !"Determine the purity of your separated samples through a melting point determination. The procedure to be used is as follows:

1. Before your practical, you should fill you should look up the safety information for the chemicals to be used, i.e.: benzoic acid, 4-nitrotoluene, diethyl ether, sodium bicarbonate solution, concentrated hydrochloric acid and ethanol from the MSDS datasheets and fill in the special assessment forms where appropriate. (see

http://staff.um.edu.mt/jgri1/safety/download).

2. To separate the benzoic acid and 4-nitrotoluene you should:

a. Place 2g benzoic acid / 4-nitrotoluene mixture in a 100mL conical flask, add 20 mL diethyl ether (no flames!) and shake. Transfer this mixture to a separating funnel and add 20mL sodium bicarbonate solution (5% by mass). This will convert the benzoic acid to the water soluble sodium benzoate.

b. Stopper the separating funnel and shake for a short while; then invert the

funnel and open the tap gently to release pressure. Close the tap and continue shaking, frequently releasing the pressure until further shaking develops little or no additional pressure. Then shake vigorously (but carefully) for 2-3 minutes to ensure the maximum possible transfer of the solutes to the appropriate phase.

c. Allow the layers to separate and run off the bottom layer into a 100mL conical flask. Then add 10mL water to the funnel, shake and run the aqueous bottom layer into the same conical flask.

3. To recover benzoic acid and to test its purity:

a. Add a few drops of concentrated hydrochloric acid to the aqueous phase until the pH is less than 2. Allow to solution to stand until crystallisation of the benzoic acid is complete.

b. Filter the solution under suction and collect the crude benzoic acid crystals in a Hirsch funnel. Wash the benzoic acid crystals twice with water and allow to drain.

c. Recrystallise from water and dry in an oven set at 105 o C.

d. Determine the mass, percentage recovery and melting point of the recovered/purified benzoic acid.

4. To recover 4-nitrotoluene and to test its purity:

a. Evaporate the ether solution to dryness over a steam bath in a fume hood, (no flames!).

b. Collect the crystals so formed and recrystallise from ethanol

c. Determine the mass, percentage recovery and melting point of the recovered/purified 4-nitrotoluene.

5. In your discussion:

a. Explain carefully the chemistry involved in the separation.

b. Discuss how would you modify the above protocol to deal with the separation of a mixture of benzoic acid and 4-nitrophenol.

University of Malta Department of Chemistry CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 5: SEPARATION INLOVING THE FORMATION OF A DERIVATIVE OF ONE OF THE COMPONENTS

Introduction

The separation of a binary mixture of substances with very similar physical properties (e.g. two similar hydrocarbons) cannot be accomplished by simple crystallisation or solvent extraction. In such cases, one needs to:

1. Chemically modify one of the components to render it significantly different from the other constituents;

2. Separate the derivative from the mixture;

3. Transform the derivative back to the original form.

This practical involves the separation of naphthalene, 1, (m.pt. 80 o C) and biphenyl, 2, (m.pt. 69 o C).

, (m.pt. 80 o C) and biphenyl, 2 , (m.pt. 69 o C). 1 2 Although

1

2
2

Although these two chemicals are extremely similar, 1 (a Lewis base) reacts with picric acid to form a picrate, 3, which 2 does not. The separation is made possible since 3 has different solubility properties from 1 and 2.

Procedure:

1.

Before your practical, you should fill you should look up the safety information for the chemicals to be used, i.e.: naphthalene, biphenyl, picric acid and concentrated ammonia and fill in the special assessment forms where appropriate. (see http://staff.um.edu.mt/jgri1/safety/download ).

2.

To form the naphthalene derivative (naphthalene picrate) and isolate it from biphenyl:

a. Place 1g of the naphthalene (50% w/w) and biphenyl mixture in a boiling tube, add 2mL of methanol and heat on a steam bath until the mixture is completely dissolved.

b. In another boiling tube dissolve 1.3g of picric acid in 10mL methanol by warming on a steam bath until a yellow solution is obtained.

c. c.

After Transfer putting the aside solution a few of drops the mixture of the picric slowly acid to solution the picric for step acid 2e, solution. to the

rest Warm of the for picric 1 minute acid on solution a steam add bath slowly and the allow solution to cool. of the This mixture will result from in

step the formation 2a. Warm of for solid 1 minute naphthalene on a steam picrate. bath and allow to cool. This will

d.

result Collect in the the crystals formation formed of solid in naphthalene a Hirsch funnel, picrate. wash with methanol, dry

d.

Collect and determine the crystals the melting formed point. in a Hirsch funnel, wash with methanol, dry and determine the melting point.

e. Check for the presence of naphthalene in the filtrate by adding a few drops of fresh picric acid solution. If no picrate forms then separation has been complete.

3. To re-extract biphenyl, purify it and test its purity:

a. Add 1mL concentrated ammonia (using a measuring cylinder) and 30mL water to the filtrate. This will result in the precipitation of biphenyl.

b. Collect the biphenyl by filtration and recrystallise from hot water.

c. Record its melting point.

4. To re-convert the picrate to naphthalene, purify it and test its purity:

a. Place the picrate crystals in a 100mL conical flask, and dissolve them in a little methanol by warming on a steam bath.

b. Add 1mL concentrated ammonia and 30mL water. Filter the naphthalene and recrystallise it from methanol.

c. Record its melting point.

University of Malta Department of Chemistry CHE 1700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 6: FRACTIONAL DISTILLATION

Introduction

Distillation is a very useful method of purifying liquids. It is used routinely for purifying solvents and reagents and, with care and the correct apparatus it can be used to separate liquids whose boiling points are only a few degrees apart.

The conventional apparatus for use in simple distillation is shown in Fig. 1a. This apparatus is suitable for distilling compounds from involatile residues, or for separating liquids whose boiling points differ by at least 50°C. The separation of liquids whose boiling points are between 5°C and 50°C apart requires the use of a fractionating column (see Fig. 1b). In its most basic form, a fractionating column may be regarded as a long vertical tube with a temperature gradient (hottest at the bottom). In practice, the use of a very long vertical tube is very inconvenient and hence it is normally replaced with a shorter ‘column’ designed to offer the same properties (e.g. the Vigreux column in Fig.

1b).

the same properties (e.g. the Vigreux column in Fig. 1b ). (a) Taken from M. Casey,

(a) Taken from M. Casey, J. Leonard, B. Lygo and G. Procter,

Advanced Practical Organic Chemistry, 1990, Blackie & Professional (UK)

Organic Chemistry , 1990, Blackie & Professional (UK) (b) Taken from A.I. Vogel, Vogel’s Textbook of

(b) Taken from A.I. Vogel, Vogel’s Textbook of Practical Organic

Chemistry, 5 th Edn., 1988, Longman (UK)

Fig. 1: (a) The setup for a simple distillation, and (b) The setup for a fractional distillation using a Vigreux column.

Fig. 2: The temperature-composition diagram, corresponding to an ideal mixture A - B where A

Fig. 2: The temperature-composition diagram, corresponding to an ideal mixture A-B where A is more volatile than component B. In fractional distillation, successive boilings and condensations of a liquid originally of composition a 1 lead to a condensate that is pure A.

Let us illustrate the process of distillation with reference to an A-B ideal * liquid mixture where A is more volatile than B (i.e. A has a lower boiling point than B, see Fig. 2). We shall assume that the initial liquid mixture has a composition a 1 and boils at a temperature of T 2 . At T 2 , an equilibrium between the liquid and the vapour phases is established, but whilst the liquid phase still has a composition of a 1 (=a 2 ), the vapour phase has composition a 2 ` which is richer in the more volatile component A. In a simple distillation, the vapour of composition a 2 ` is withdrawn and condensed. Thus, in this example, the first drop of the condensate would have given a liquid of composition

(the same as a 2 ` , i.e. a mixture which is richer in the more volatile component, A, than the

original liquid mixture of composition a 1 ).

a

3

In fractional distillation, the vapour of composition a 2 ` is not collected, but it is allowed to cool slowly as it ascends the fractionating column, until eventually it returns to the condensed phase of composition a 3 (the same composition as the vapour a 2 `). However, as this condensate descends the column, it exchanges heat with the ascending hot vapour and the vapour is enriched with the more volatile component at the expense of the liquid, in an attempt to reach equilibrium within the liquid-vapour system. These boiling- condensation cycles are repeated successively until ‘almost pure’ A is obtained at the top of the column.

* In reality, some substances do not form ideal solutions, and non-ideality will lead to the formation of azeotropic mixtures, which results in discrepancies from the theory described here. For a discussion on distillation of non-ideal solutions, please refer to standard physical chemistry textbooks, such as P.W. Atkins, Physical Chemistry, 6 th Edn., 1998, OUP (UK), Chapter 8. At a given pressure, the boiling point is the temperature when the liquid is in equilibrium with the vapour.

All this suggests that the conditions necessary for a good separation are:

o

Comparatively large amounts of liquid continually returning through the column;

o

Thorough mixing of liquid and vapour;

o

A large active surface of contact between liquid and vapour.

This means that cooling in the fractionating column must occur very slowly. Excessive cooling (e.g. because of drafts) should be avoided by, for example, insulating or lagging the outer surface of the column or, if possible, by surrounding it with a vacuum jacket or an electrically heated jacket. This difficulty is particularly important when separating liquids of high boiling points.

It is important to ensure that the procedure is carried out safely. In particular:

1. Never heat a closed system. If you are worried about moisture entering your system, you can attach a drying tube to the vents.

2. Take special care when distilling organic solvents to avoid fires and explosions. Therefore:

a. Ensure that the vapour does not come into contact with flames, sources of sparks (electrical motors), or very hot surfaces (hot plates).

b. Do not heat (especially round bottomed flask) using a Bunsen burner – Always use an electrically heated oil/water bath or a heating mantle.

c. Never allow a distillation pot to boil dry. The residues may ignite or explode.

d. Beware of the possibility that ethers and hydrocarbons may be

contaminated with peroxides that form on exposure with air.

e. Always check whether any of the liquids you plan to distil are thermally unstable.

3. Always add some boiling chips (or if possible, stir the liquid using a magnetic stirrer), in order to prevent bumping.

Procedure:

In this practical, you have shall be separating the two organic liquids, acetone and toluene from a 1:1 mixture of the two, determine their boiling point, and determine which of the two is the carbonyl compound (acetone) through the 2,4-dinitrophenylhydrazine (2,4- DNPH) and the iodoform tests.

1. Before your practical, you should fill you should look up the safety information for the chemicals to be used and fill in the special assessment forms where appropriate. (see http://staff.um.edu.mt/jgri1/safety/download). You should also look up the boiling points of pure acetone and toluene.

2. Separate about 50mL of an acetone/toluene mixture and measure the boiling points of acetone and toluene:

a. Set up the apparatus for fractional distillation as in Fig. 1b using a 100mL round-bottomed flask and the column provided (insulated with cotton wool). Note that (1) the bulb of the thermometer should be just below the

level of the side arm, (2) a very small piece of glass wool (not cotton wool) should be used in the fractionating column to support the glass beads.

b. Place about 50mL of the acetone/toluene mixture in the 100mL round- bottomed flask together with a few boiling chips (porous pot etc.).

c. Heat the flask slowly until the reading on the thermometer reaches a steady state (the boiling point of the first fraction) and drops are observed to condense out of the Leibig condenser . Record this steady state temperature and collect the distillate in a conical flask. Allow the distillation to proceed until no more liquid gets out of the condenser/measuring cylinder. Record the volume of the first fraction.

d. When all of the first fraction has been distilled, the temperature at the top of the column will be observed to increase and then reach a second steady state (the boiling point of the second fraction) and whist drops of the second fraction will start to condense out of the leibig condenser . Record this second steady state temperature (the boiling point of the second fraction) and collect the distillate in a clean conical flask/measuring cylinder § . You should collect as much of the second fraction as practically possible making sure that you so not allow the distillation pot to boil dry. Record the volume of the second distillate.

3. Identify which of the two liquid distillates has a carbonyl group (acetone) using the 2,4-DNPH and iodoform tests:

a. For the 2,4-DNPH test: Add 2-3 drops of the liquid to be tested to 3mL of 2,4-dinitrophenylhydrazine and shake. If no precipitate forms immediately, allow to stand for 5-10 minutes. Record the observations and inferences in your laboratory notebook.

b. For the iodoform test: Dissolve 4 micro drops of the liquid to be tested in water (2mL). Add 10% sodium hydroxide (2mL), and then slowly add 2mL of the iodine solution. If the substance is insoluble in water, dissolve it in dioxane (2mL). Proceed as above and at the end dilute with water (10mL). Record the observations and inferences in your laboratory notebook.

You should collect in a separate container (and then discard it), any liquid collected (i) before the boiling point of the first distillate, and (ii) as the temperature is rising between the boiling point of the first fraction and that of the second fraction. § Since you have a binary A-B mixture, the second main distillate can be obtained without the use of a fractionating column.

University of Malta Department of Chemistry CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 7/8: THE OXIDATION OF CYCLOHEXANOL TO CYCLOHEXANONE & ITS EXTRACTION THROUGH STEAM DISTILLATION.

Introduction

This practical involves:

i. The oxidation of cyclohexanol to cyclohexanone using acidified sodium dichromate;

ii. The extraction of the cyclohexanone so formed from the rest of the reaction mixture though steam distillation (the distillate is a water/cyclohexanone mixture);

iii. The extraction of cyclohexanone from the water/cyclohexanone mixture in ether*;

iv. Simple distillation to purify the cyclohexanone (the boiling points of ether and cyclohexanone are very different)*.

(* To be performed in the second week)

Steam distillation is a technique where a water immiscible substance A with a high boiling point can be ‘distilled’ with the help of steam at a temperature that is lower than 100 o C.

A substance A is immiscible with water (substance B) when the mutual solubilities of A and B are very small. At equilibrium, an A/B system will consist of two liquid phases and a single gaseous phase, i.e.:

(1) A liquid phase containing mainly A which is saturated with B (the low solubility of B in A means that this phase is almost pure A); (2) A liquid phase containing mainly B which is saturated with A (the low solubility of A in B means that this phase is almost pure B); and (3) A vapour phase where A and B coexist (A and B are miscible in the gaseous phase). The total vapour pressure, p, for this A/B system is approximately given by:

pp=

*

A

+ p

*

B

are the vapour pressures of pure A and pure B. These vapour pressures

i * increase as T increases (volatility increase

with an increase in temperature). The system will start boiling i (i.e. distilling) at the point when the total vapour pressure becomes equal to the atmospheric pressure, i.e. p = p atm .

, the boiling

are functions of temperature, T, such that

where

p

*

A

and p

*

B

p

This occurs at a temperature

boil

T

A-B

that is less than either of

boil

T

A

or

boil

T

B

points of pure A and B respectively ii .

i Boiling can be considered as the point when the dissolved substances are purged out of their solution. ii For boiling to progress sucessfully at a single temperature, there must be continuous presence of the saturated solutions ‘A in B’ and ‘B in A’. This ‘continuous presence’ is ensured at all times since these very dilute solutions can be very quickly replenished to their saturatiuon composoition aided by the vigorous agitation induced by boiling.

For

temperature reaches

example,

assuming

boil

T

B

that

boil

T

B

boil

< T

A

,

boiling

(=100 o C if B is water) since

***

pT

boil

+ pT

boil

()

AB

()

BB

= pT

AB

()

boil

+>p

atm

p

atm

of

A/B

pT

B

B

*

(

boil

)

must

= p

atm

occur

before

i.e.:

the

There is however one important snag to this technique, namely that the composition of the vapour (and hence of the distillate) is in proportion to the vapour pressures of the components, i.e.:

n

A

p

*

A

(

boil

T

A

)

=

n

B

p

*

B

(

boil

T

B

)

This means that substance of a high boiling point with a low volatility will distil in low abundance.

To illustrate this we shall look at a nitrobenzene (=A) and water (=B) mixture. These

substances boil (i.e. their respective vapour pressures reaches

211 o C (i.e.

= 372.4 K ), as illustrated in Fig. 1. This

results in a reduction of the boiling point of approximately 111K. However, at 99.4 o C,

mixture of the two boils at 99.4 o C (i.e.

= 373 K ) respectively, but a

760 mmHg ) at 210-

p

atm

=

boil

T

A

=

483

484 K )

and 100 o C (i.e.

boil

T

A-B

boil

T

B

p

*

A 20 mmHg and p

*

B

740 mmHg , i.e.:

n

A

p

*

A

(

boil

T

A

)

=

n

B

p

*

B

(

boil

T

B

)

≈= 20

740

1

37

i.e.

w

A

n

A

(

× RMM A

)

=

w

B

n

B

×

(

RMM B

)

1

×

123

≈=

×

37

18

123

666

i.e. the wt% of nitrobenzene in the distillate will only be:

wt% =

w

A

w

A

+

w

B

×≈ 100%

15%

Nevertheless, the low wt% of nitrobenzene in the distillate is by far overweighed by the fact that the distillation could be carried out at 99.4 o C rather than at the boiling point of nitrobenzene, which is 210-211 o C.

Note that the technique of steam distillation is very important in natural product chemistry where it is employed on a industrial scale to extract essential oils from plants iii .

iii See, for example: ‘A Guide to Aromatherapy’ web-site, http://www.fragrant.demon.co.uk/, for a list of oils used in aromatherapy and their method of production, ‘The English Chamomile Company’ web-site, http://www.chamomile.co.uk/ for a discussion of steam distillation in the extraction of chamomile, or http://www.distillation.co.uk/, a web-site for a company which specialises in the production of distillation equipment for use in the production of essential oils.)

Fig. 1: The vapour pressure v s . temperature curve for a nitrobenzene-water system. Note

Fig. 1: The vapour pressure vs. temperature curve for a nitrobenzene-water system. Note that whilst pure nitrobenzene boils at 483-484K, the when it is mixed with steam, it will boil at the much lower temperature of 372.4K when the vapour pressure of the nitrobenzene-water system reaches 760 mmHg (atmospheric pressure).

Procedure:

(a) Before your practical, you should look up the risks/safety information for the known chemicals used in this practical. (b) In the first session of your practical you will be oxidising the cyclohexanol to cyclohexanone and extracting the cyclohexanone by steam distillation. 1) At the beginning of the practical, fill the steam generator (to be used for the steam distillation) with water (3/4 full) and heat it with a Bunsen burner. (Boiling of such a quantity of water requires a plenty amount of time!)

2)

To oxidise the cyclohexanol to cyclohexanone:

i. Dissolve 15g sodium dichromate dihydrate in 25mL 2M H 2 SO 4 by heating the mixture in a beaker. Once all the sodium dichromate dihydrate dissolves, cool the mixture to 15 o C by immersing the beaker in ice.

ii. Prepare a mixture of 15g cyclohexanol in 10mL 2M H 2 SO 4 (10mL).

Cool in ice to below 15 o C. iii. Add the dichromate onto the cyclohexanol and stir. Note that the reaction is exothermic, and you will observe that the temperature starts to rise very rapidly. This increase in temperature is beneficial

as it increases the rate of the oxidation reaction. However, you should not allow the temperature to exceed 60 o C.

iv. Keep the reaction going until the temperature begins to drop and the solution turns green, indicting that the reaction is ‘complete’. (This should take about 15 minutes from the addition of the start of the reaction). Allow the reaction mixture to stand for another 10 minutes.

3)

To extract the cyclohexanone so formed by steam distillation:

i. Pour the solution into a 250mL two-necked round-bottomed flask. Rinse the reaction flask with water (100mL, split between three rinses) and add the washings to the distillation flask.

ii. Set up the steam distillation apparatus according to the instructions given by your demonstrator.

iii. In theory, the steam generated from the steam generator should be enough to heat the reaction mixture. In practice, this would take a long time, and hence you are advised to accelerate the process by heating up the round bottomed flask directly until the reaction mixture starts boiling. The steam distillation should then be allowed to proceed on its own.

iv. When about 50mL of distillate is collected, add another 50mL of distilled water to the round bottomed flask and continue distilling until no more oil passes over with the water. This is an indication that the steam distillation is ‘complete’.

v. Cover the distillate, label it, and store it in your laboratory cupboard for the next session.

(c) In the second session of your practical you will be extracting the cyclohexanone from the cyclohexanone-water distillate, and further purifying it through simple distillation.

1)

To extract the cyclohexanone from the distillate:,

i. Salt out the distillate by adding sodium chloride (approx.

25mL 20mL

of

0.2g/mL solution) and extract with ether (25mL).

ii. Wash the ether layer with 10% sodium hydroxide (25mL) and then re-wash with the salt solution (approx. 25mL of 0.2g/mL solution).

iii. Dry the ethereal extract over anhydrous sodium sulphate and filter into a distillation flask.

2)

To purify the extracted cyclohexanone:,

i. Set up the simple distillation apparatus according to the instructions given by your demonstrator.

ii. Distil the ether over a steam bath into a Buchner flask having a side- arm fitted with a rubber tubing leading well below the level of the bench. This precaution is essential when ether is being distilled because of the risks of fire and explosion (Air and ether vapour form an explosive mixture when the latter reaches a flame.).

iii. Once all the ether has been distilled off, heat the remaining mixture with a heating mantle to distil the cyclohexanone.

iv. Record the temperature of collection and the yield in grams and in moles. Calculate the percentage yield.

University of Malta Department of Chemistry CH E1700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 9: DETERMINATION OF THE COMPOSITION OF A ‘BORAX / BORIC ACID' AQUEOUS SOLUTION.

Introduction

In this practical you shall be measuring quantitatively the concentration of boric acid and borax in a boric acid / borax solution through acid/base titrations.

Boric acid, 1, is too weak an acid to be titrated quantitatively in an aqueous medium using a visual indicator. However, it can be titrated with standard alkali (NaOH) in the presence of certain polyols (e.g. propane-1,2,3-triol, 2, commonly known as glycerol), using phenolphthalein as indicator.

This is because 1 and two molecules of 2 can form a complex, (borospiranic acid, 3, which may be regarded as a diester of glycerol) in which two rings are attached to the boron atom, in mutually perpendicular planes.

to the boron atom, in mutually perpendicular planes. (eqn. 1) Borospiranic acid is a strong enough

(eqn. 1)

Borospiranic acid is a strong enough acid to give a satisfactory end-point with strong aqueous alkali (e.g. NaOH) using phenolphthalein as indicator (see Appendix 1). Note that the overall stoichiometry of the reaction can be represented as follows (eqn. 2):

(

B OH

)

3

+ NaOH

→ NaBa OH

glycerine

(

)

4

(eqn. 2)

(Question: Given that the borospiranic acid is a monobasic acid, which is the labile proton in the structure? Give a reason for your choice.)

The propable 3D structure of the proposed borospiranic acid complex, 3 . The sodium salt

The propable 3D structure of the proposed borospiranic acid complex, 3.

The sodium salt of boric acid, borax (4, Na 2 B 4 O 7 .10H 2 O, also known as sodium tetraborate decahydrate) is a weak base. It can be quantitatively titrated directly with standard acid (e.g. HCl) using methyl red as an indicator (see Appendix 1) as in eqn. 3:

Na B O .10H O + 2HCl 4H BO

247

2

3

(4)

(1)

3

+ 2NaCl + 5H O

2

(eqn. 3)

One should note that these two determinations are independent of each other, and hence, if performed carefully, they can be used to quantitatively measure the amounts of boric acid and borax present in a mixture of the two.

Procedure:

In this practical you shall be applying the theory described above to measure quantitatively the concentration of boric acid and borax in a aqueous solution containing both boric acid and borax solution. However, you shall first be practicing the titrations on the two components present on their own, and at the same time, use these ‘practice titrations’ to determine quantitatively the mass percentage (i.e. percentage purity) of the boric acid and borax as supplied.

(a) Before your practical, you should look up the risks/safety information for the known chemicals used in this practical. (b) To determinate of the mass percentage of boric acid: Accurately weigh about 1g of sample and transfer it to a clean conical flask together with 50mL water. Carefully boil the solution for about 1 minute so that the boric acid dissolves completely and any carbon dioxide in the distilled water is completely exsolved. Be careful not to loose any solute during boiling. Cool, add about 15mL glycerol, and titrate with (about) 1M NaOH using phenolphthalein as indicator. (The actual molar concentration will be provided during the practical.) Check whether the titre values that you obtain conform to

the amount of boric acid used. Repeat this procedure two more times. Use the results of the titration to calculate the mass percentage (i.e. purity by weight) of the boric acid as supplied. (c) To determine the mass percentage of borax: Dissolve about 1.2g of borax (accurately weighed) in about 10mL water, and heat to help dissolution. Cool and titrate with (about) 0.25M HCl using methyl red as indicator. Check whether the titre values that you obtain conform to the amount of borax used. Repeat this procedure two more times. Use the results of the titration to calculate the mass percentage (i.e. purity by weight) of the borax as supplied. (d) To determine the composition of an aqueous solution labeled M containing borax and boric acid: Pipette 25mL of M into a conical flask and titrate with (about) 0.25M HCl using methyl red solution as indicator. Record the volume of acid required. Reserve the titration liquid and repeat this portion of the assay to obtain concordant results. This should give you a measure of the amount of borax present. To determine the amount of boric acid, boil the titration liquid to expel carbon dioxide. Cool the solution, add glycerol (15mL), and titrate with (about) 1M NaOH using phenolphthalein as indicator, ensuring that you obtain concordant results. Use these measurements to calculate the concentration of borax and boric acid in the solution mixture M.

Note: You are expected to provide a quantitative estimate of the errors involved in these determinations. A discussion on ‘Errors in Quantitative Analysis’ is provided on http://staff.um.edu.mt/jgri1/teaching/ch130/errors.pdf .

Appendix 1:

pH Transition ranges and colour of some common indicators *

pH Transition ranges and colour of some common indicators * * Taken from G.D. Christain, Analytical

* Taken from G.D. Christain, Analytical Chemistry, 4th Ed., John Wiley & Sons, USA,

1996.

University of Malta Department of Chemistry CH E1700 - Chemistry Practical I - B.Sc. (Hons.)

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 10: PERMANGANATE METHODS

Introduction

Redox (reduction/oxidation) titrations are extremely important in analytical chemistry. In these reactions, one reactant (the oxidising agent or oxidant) will have a stronger affinity for electrons than a second species (the reducing agent or reductant) and will tend to extract electrons from it.

It is convenient to analyse a redox reaction in terms of two half-reactions, one which describes the oxidation, and one which describes the reduction. For example, the equation for the oxidation of iron(II) ions by permanganate ions in an acidic solution (eqn. 1) may

Mn + (eqn.

2), and (ii) the oxidation of Fe 2+ to Fe 3+ (eqn. 3). These two half reactions show very clearly that the permanganate ion is the electron acceptor (i.e the oxidising agent) whilst the iron(II) ion is the electron donor (i.e. the reducing agent):

be viewed in terms of the two half-reactions: (i) the reduction of

MnO to

4

2

The full reaction:

5Fe

2+

+ MnO + 8H

-

4

+

5Fe

3+

+ Mn

2+

+ 4H O

2

(eqn. 1)

The corresponding half reactions:

MnO

5Fe

4-

+ 8H +5e

5Fe

3

+

2

+

+−

+

5e

Mn

2+

+4H O

2

(eqn. 2)

(eqn. 3)

The ability (potential) of a substance to act as an electron acceptor (i.e. as oxidising agent) can be measured in terms of the magnitudes of the standard reduction potentials, E 0 , (sometimes referred to as standard electrode potentials) of the half reactions. The higher the standard reduction potential of a species, the better is its ability to act as an oxidising agent (i.e. be reduced).

Standard reduction potentials are always quoted relative to the half reaction where two hydrogen ions accept two electrons to form a hydrogen molecule which is arbitrarily given a standard reduction potential of 0V (see Appendix 1). Tables of standard potentials can be found in reference books or physical /analytical chemistry textbooks.

One should note that the standard reduction potential measures the relative intensity of the driving force for the reduction half reaction. This means that, for example, the half

reactions

reduction potential, E 0 = 0.771V. Thus referring to Appendix 1 and by re-writing eqns. 2 & 3 as reductions with their associated standard reduction potentials, we may observe that:

both have the same standard

Fe

3

+−

+

e

Fe

2

+

and

5Fe

3

+−

+

5e

5Fe

2

+

MnO + 8H +5e

5Fe

-

4

+

5Fe

2

+

3

+−

+

5e

Mn

2+

+4H O

2

E 0 = 1.51V E 0 = 0.771V

E 0 = 1.51V E 0 = 0.771V

(eqn. 4)

(eqn. 5)

that is, the standard reduction potential of the permanganate ion is higher (more positive) than that of the iron(II) ions, and hence, the forward of eqn. 4 and the reverse of reaction eqn. 5 will take place, as indicted in eqns. 1-3. Note that for a redox reaction to go to completion there must be a difference in the standard reduction potentials of the two half reactions of at least 0.1V. This is an important requirement if a redox reaction is to be used in quantitative analysis.

This practical involves a number of titrations against potassium permanganate, KMnO 4 , namely:

i.

Oxalic acid against KMnO 4 to standardize the KMnO 4 solution;

ii.

Iron(II)sulphate against KMnO 4 to determine the concentration of Fe ++ ;

iii.

Hydrogen peroxide against KMnO 4 to determine the volume strength of the peroxide solution.

The standard potentials for the half reactions involved in these process (taken from Appendix 1) are as follows:

(1)

(2)

(3)

(4)

E

θ

, V

MnO + 8H +5e

2CO

Fe

O

+

2

-

4

+

C O

24

2

+

Mn

2+

+4H O

2

1.51

3

2

+

2e

+−

+

e

2H

Fe

2- -0.440

0.771

0.682

+

+→ 2e

H O

22

Since the standard reduction potential of the half reactions (2)-(4) are lower than that of (1) (i.e. the permanganate has the greatest tendency to be reduced), the reverse of reactions (2)-(4) occur in these titrations. This means that in all of these reactions, the potassium permanganate is acting as the oxidising agent. Note that in all of these reactions, there is no need of an indicator since a 0.02M solution of potassium permanganate is deep purple whilst the product of its reduction, Mn 2+ , is nearly colourless. Thus, before the endpoint, the purple colour of the permanganate ion is removed practically as soon as it is added (see below) whilst as soon as the titration is complete, a fraction of a drop of excess permanganate will impart a definite pink colour to the solution.

The mechanism of the reactions between oxalate and permanganate ions is extremely complicated. The first few drops of permanganate react slowly with the oxalic acid but after a small amount of manganous salt has been formed, the reaction occurs almost instantaneously in hot solution. It appears that the Mn 2+ ions catalyze the reaction between permanganate and oxalic acid. This can be verified by adding a small amount of manganese(II) salt at the very start of the titration. The permanganate is then reduced rapidly from the start.

Although the titration of oxalic acid (obtained by dissolving sodium oxalate (Na 2 C 2 O 4 ), previously dried at 100-110 o C) against potassium permanganate is a standard technique to standardise permanganate solutions, special care must be taken since hot solutions of oxalic acid decompose slowly:

H C O

224

CO

2

+ CO + H O

2

This decomposition reaction is catalysed by manganous salt and hence the titration should not be carried out too slowly as this could lead to an error (a too low titre value).

However, if the titration is too rapid, or there is insufficient stirring, the added permanganate will tend to spend a longer time than necessary in the presence of hot acid. This will result in the auto-decomposition of the still unreacted permanganate with the evolution of oxygen, and this will be reflected in a too high titre value. (Note that if the titration is carried out according to the standard procedure, these two side reactions will only occur to a negligibly small extent.)

Hydrogen peroxide solution is an aqueous solution of hydrogen peroxide containing not less than 5% and not more than 7% w/v of H 2 O 2 , corresponding to about 20 times its volume of available oxygen. In the titration of hydrogen peroxide with permanganate, we observe a similar phenomenon as with oxalic acid, that is, the first few drops of permanganate are decolourised slowly but again the reaction is catalysed by adding a trace of a manganous salt at the very beginning of the titration.

Procedure:

(a)

Before your practical, you should look up the risks/safety information for the known chemicals used in this practical.

(b)

Preparation of approximately 0.02M KMnO 4 :

Dissolve 0.8g KMnO 4 in distilled water and make up to 250mL. Transfer to a beaker and heat to about 90 o C for 15 minutes. Use after cooling to room temperature.

(c)

Standardisation of potassium permanganate:

i. Prepare an approximately 0.04M sodium oxalate solution by dissolving about 0.54g of sodium oxalate (Na 2 C 2 O 4 , previously dried at 100-110 o C) in distilled water and making up to 100.0mL (using a 100mL volumetric flask). Calculate the accurate concentration of the sodium oxalate solution.

ii. Pipette 25mL of the 0.04M sodium oxalate solution to a 250mL conical flask and add 50mL 2M sulphuric acid.

iii. Heat to about 80 o C and titrate slowly with permanganate whilst shaking the flask well. If the solution remains pink after adding the first drop or two, wait until it clears before continuing the titration.

iv. Continue the titration slowly until a faint permanent pink tinge is obtained. The temperature should not be less than 60 o C at the end-point. If it falls below this during the titration, the solution must be reheated.

v. Repeat this process twice more, or until you obtain concordant results to obtain a mean value for the molar concentration of the permanganate. Calculate the molar concentration of the potassium permanganate.

(d)

Determination of the percentage of iron(II) in iron(II)sulphate crystals using standardised potassium permanganate solution:

i. Weigh out accurately about 3.5g of iron(II)sulphate crystals. Dissolve, and make up to 100.0mL using a 100mL volumetric flask using 2M sulphuric acid.

ii. Pipette 25.0mL of iron(II)sulphate solution into a 250mL conical flask, add 25mL 2M H 2 SO 4 and titrate with KMnO 4 solution to the first permanent pink end-point. Repeat the titration two or three times.

iii. Calculate the percentage of ferrous iron in the crystals.

(e) Determination of the concentration and volume strength of a peroxide solution using standardised potassium permanganate solution

i. Dilute 10.0mL of the hydrogen peroxide solution (measured using 10mL pipette or a burette) to 250mL with water in a volumetric flask. ii. Pipette 25.0mL of the diluted solution to a conical flask, add 5mL of sulphuric acid (50% w/v) and titrate with the potassium permanganate solution to the first permanent pink end-point. Repeat the titration two or three times until you obtain concordant results. iii. Calculate the molar concentration and the ‘volume strength’ of the original peroxide solution.

Appendix 1:

Standard potentials at 298K in electrochemical order *

1: Standard potentials at 298K in electrochemical order * Others † : 2CO 2 + 2e

Others :

2CO

2

+

2e

C O

24

2-

E 0 = -0.440 V

* Taken from P.W. Atkins, Physical Chemistry, 6th Ed., OUP, UK, 1998.

Taken from G.D. Christain, Analytical Chemistry, 4th Ed., J. Wiley & Sons, Inc., USA, 1986.

University of Malta Department of Chemistry CH E 1 70 0 - Chemistry Practical I

University of Malta Department of Chemistry

CHE1700 - Chemistry Practical I - B.Sc. (Hons.) I Year

EXPERIMENT 11: DETERMINATION OF WATER HARDNESS

Introduction

This practical involves:

(i)

The determination of the total, permanent and temporary hardness of two different samples of tap water through complexometric titrations;

(ii)

The determination of the total calcium and magnesium content of two different tap water samples through complexometric titrations;

(iii)

The assessment whether the two water samples are significantly different in terms of their hardness.

(a) Water hardness

Water hardness is a measure of the mineral levels (usually calcium and magnesium salts) dissolved in the water. Hard water requires more soap and synthetic detergents i and contributes to scaling in boilers and other industrial and domestic equipment. Beacuse the concentrations of calcium and magnesium ions in natural water are usually much higher than the concentrations of the other ions, it is common practice to measure water hardness by measuring the concentrations of Ca ++ and Mg ++ .

Hardness can either be temporary or permanent, this being determined by the anions associated with the Ca 2+ /Mg 2+ cations. If the anion is a carbonate or bicarbonate, the hardness is temporary and can be removed by boiling the water and allowing it to cool. Sulphates and chlorides result in permanent hardness that cannot be removed by boiling. This type of hardness will not contribute to scaling but will cause waste of soap and detergent. (Temporary hardness that will contribute to scaling and soap precipitation).

Hardness is measured as the equivalent to the amount of calcium carbonate (CaCO 3 ) in water. It is measured in mg L -1 (milligrams per litre) or as ppm (parts-per-million) CaCO 3 (even though all the hardness may not be due to CaCO 3 ). Various international agencies have produced guidelines for water hardness classification. For example, the United States Environmental Protection Agency (EPA) classification for total hardness is as follows:

Equivalent CaCO 3 content (ppm or mg L -1 )

Description

0-17.1

Soft

17.2-59.9

Slightly hard

60-128.2

Moderately hard

128.3-179.6

Hard

i Historically, water hardness was understood to be a measure of the capacity of the water for precipitating soap. Soap precipitaion occurs in the presence of polyvalent metals such as calcuium, magnesium, aluminum, iron, manganese, strontium and zinc, and by hydrogen ions.

179.7+

Very Hard

(b) Complexometric titrations: The quantitative analysis of water hardness

Cations (acting as Lewis acids) can form complexes with substances that have a pair of unshared electrons (known as ligands, i.e. molecules with the atoms such as N, O and S

that have a lone pair and can act as Lewis bases) provided that the Lewis base can satisfy

the coordination number of the Lewis acid cations. The number of ligand molecules that

can

attach to a single cation will depend on (i) the coordination number of the metal ion,

and

(ii) the number of complexing groups on the ligand molecules. An organic molecule

that

has two or more groups capable of complexing with a metal ion is known as a

chelating agent and the resulting complex is known as a chelate.

The formation of metal complexes can form the basis of quantitative analysis through

titrations (complexometric titrations) for a quantitative determination of the amount of

metal present in a sample. Furthermore, these methods and are particularly useful when the concentrations of these metal ions are present at the millimole level ii .

A chelate

ethylenediaminetetraacetic acid, EDTA the structural formula of which is as follows:

that

is

very

commonly

used

in

complexometric

titrations

is

EDTA is a tetraprotic acid and commonly represented as H 4 Y where the four hydrogens

represent the four ionisable hydrogens on the four carboxyl groups. Since H 4 Y has a very

low solubility in water, the disodium salt Na 2 H 2 Y.2H 2 O is more commonly used. The pH

used for the reaction must also be carefully chosen and controlled (see standard analytical chemistry textbooks for details).

The determination of the hardness due to Ca 2+ and Mg 2+ can be carried out at pH 10

(controlled using an ammonium chloride / ammonium hydroxide buffer) using the indicator Eriochrome Black T. Eriochrome Black T is itself a chelating agent having three ionisable protons (hence representable by H 3 In) which can complex with Mg 2+ to

from the red complex MgIn . iii

When EDTA (in the from of H 2 Y 2- ) is added to a sample containing free Ca 2+ and Mg 2+

and MgIn , the EDTA will first complex with all the free Ca 2+ , then with all the free

Mg 2+ , and finally, once all the Ca 2+ and Mg 2+ free ions are used up, the strong EDTA ligand displaces indicator from the magnesium. This gives rise to uncomplexed indicator HIn which has a very distinctive blue colour:

2

MgIn

(red)

+

H Y

2

2

(colourless)

MgY

22

++ HIn

−+

H

(colourless)

(blue)

Note that this colour change from red to blue (titration end-point) can be made sharper if the indicator is kept as dilute as practically possible.

ii Note that similar titrations can be carried out for the quantitative determination of anions, e.g. the determination of chlorides through argenometric methods. iii Note that the calcium-indicator complex is less stable than the magnesium-indictor complex, and if no Mg 2+ is present in solution, then Eirochrome Black T cannot be used to give a sharp end-point. If no Mg 2+ is present, a few drops of Mg-EDTA (Na 2 MgY(aq)) can be added to obtain a detectible end-point.

This titration can be carried out on tap water ‘as supplied’ to determine the total hardness of the water. It may also be carried out on water that has been previously boiled for 20-30 minutes (after it has been cooled, filtered and made to volume, see procedure below) to determine the permanent hardness. (Boiling precipitate the carbonates and bicarbonates as CaCO 3 and MgCO 3 .) Furthermore, it the titration is carried out at higher pH (12), then the Mg 2+ would not contribute to the titre value as at this high pH, Mg 2+ precipitates as Mg(OH) 2 . The end point in such determinations may be carried out using a suitable indicator, e.g. Patton and Reeder’s indicator or calcon.

(c) Statistical testing for differences between means

Chemists are often entrusted with the task of assessing whether the chemical composition (or more simply whether the concentration of some particular component) of two different samples are the same or not. Because of experimental error, such an assessment needs to be done using the appropriate statistical tool.

In this practical you are required to establish whether the total hardness of the two water samples as supplies are statistically different or not. Let us assume that on the first sample (sample A), you have carried out N A titrations against EDTA to give titre values

, and that on the second sample (sample B) you have carried out N B

titrations against EDTA to give titre values

xx ,

A,1

A,2

,

,

x

A, N

A

xx ,

B,1

B,2

,

,

x

B, N

B

.

You may recall that a good estimate of the true mean titre value at 95% confidence limit for sample A (and similarly for sample B) is given by iv :

where

µ

A

x

A

=

t s A A ± x A N A
t
s
A
A
±
x A
N
A

is the arithmetic mean of the titre values

xx ,

A,1

A,2

,

+

xx

++

x

x

=

A,1

A,2

A, N

A

A

N

A

s A is the standard deviation of the titre values

s

A

=

N A 2 ∑ ( x − x ) A, i A i = 1
N
A
2
∑ (
x
− x
)
A,
i
A
i = 1
N
− 1
A

xx ,

A,1

A,2

,

,

x

A, N

A

,

x

A, N

A

given by:

given by:

and t A is a statistical factor (the t-value) which depends on N A and the confidence level desired. This factor may be obtained from standard statistical tables (see appendix 1) where it is usually quoted in terms of ν and α, where ν is the number of degrees of freedom given by ν = N A – 1, and α relates to the confidence limit (The percentage confidence limit is given by (1 – 2α)100%. Thus for example, a 95% confidence limit is equivalent to an α value of 0.025). Fore example if N A = 3 (i.e. ν = 2), then the t-value at a 95% confidence level (i.e. α = 0.025) is 4.303 (see Appendix 1).

iv These equations are valid for N A < 30.

The test to check whether the computed average titre values of two samples are statistically different is based on this equation of µ. A pooled standard deviation, s p , for the N A + N B measurements is calculated from the two standard deviations s A and s B for the two samples. This is given by:

s

p

=

2 2 Ns + Ns AA BB N + N − 2 A B
2
2
Ns
+ Ns
AA
BB
N
+ N
− 2
A
B

We can that say at 95% confidence limit, there is no significant difference between the hardness of the two samples if:

N + N A B x − x < ts A B p N N
N
+ N
A
B
x
− x
< ts
A
B
p N
N
A
B

where t is obtained from statistical tables at α = 0.025 and ν = N A + N B – 2.

For example, if N A = N B = 3, the number of degrees of freedom of the spooled data sample is given by ν = 3+3-2=4, i.e. at 95% confidence limit (α = 0.025), the statistical factor t will have a value of 2.776. Thus, according to the experiments performed, the two samples A and B do not have a significant different ammount of hardeness (at 95% confidence limit) if:

i.e.

3 + 3 xx − < 2.776 s A B p 3.3 xx − <
3
+ 3
xx
<
2.776 s
A
B
p
3.3
xx
<
2.267 s
A
B
p

and a different amount of hardness (i.e. a difference in the hardness which is higher than what can be expected from random errors at 95% confidence limit) if:

xx − A B
xx
A
B

>

2.267 s

p

Note that a higher confidence level (i.e. a smaller value of α) for the same value of ν will result in a higher value of t. For example, at 99% confidence level (α = 0.005), the hypothesis that samples A and B do not have a significant different ammount of hardeness

p , i.e. a considerable

p at 95% confidence limit. This is

relaxation from the requirement that

(at 99% confidence limit) will hold provided that

xx − A B
xx
A
B

< 3.759 s

xx − A B
xx
A
B

< 2.267 s

because the acceptable range of values where we can be sure that at 99% confidence level

is solely due to random errors will be higher than if we only want to be sure at a

95% confidence level.

x − x A B
x
− x
A
B

An alternative approach for this analysis is to compute a value of