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Cleapss Recipe Book
Cleapss Recipe Book
CLEAPSS
Recipe Book
Risk Assessment
Where a risk assessment includes control measures these have been incorporated into the instructions,
as describe in our Guidance Leaflet G90, Making and Recording Risk Assessments in School Science.
Each recipe therefore includes model risk assessments but does not include factors that you have to
thinks about, such as technician and teacher experience, and prep room conditions.
As in the previous edition, there is no mention of general bench solutions. In the past such solutions
were often much more concentrated than required and posed unnecessary hazards and risks. The
principle we follow (as in COSHH) is that the concentration of any reagent should be the lowest at
which the procedure works satisfactorily to give the intended result. Sometimes, therefore, we suggest
solutions which may appear to be an odd concentration but this nevertheless is the most suitable. For
example, 0.4 M sodium hydroxide solution is IRRITANT, whereas 0.5 M is CORROSIVE.
Making solutions
The recipes also make use of the laboratory jug as a measuring tool. Although apparently not
particularly accurate, we routinely achieve concentrations of between 1.95 and 2.05 M when using a jug
to prepare 2M sulfuric(VI) acid from concentrated sulfuric(VI) acid during the Practical Techniques in
Chemistry course. Clearly, using a jug can produce solutions with concentrations that are sufficiently
accurate for many laboratory purposes.
You will find more detail about making solutions in section 7.6 of the Handbook. If you run into
difficulties not covered in either the Recipe Book or the Handbook, phone CLEAPSS on 01895 251496
but do try the index first.
Contents
Recipe sheet
Number
Agar
Alcohol/water and
propanone/water solutions
Alginate beads
Recipe sheet
Number
30
Copper(II) solutions
31
32
Aluminium solutions
33
Ammonia gas
2,4-Dinitrophenylhydrazine
solution
3,5-Dinitrosalicylic acid
34
35
Ammonium chloride
Electroplating solutions
36
Enzymes
37
Azo dyes
Etching solutions
38
Barium solutions
10
Ethanoic acid
39
11
Fehlings solutions
40
12
41
13
14
42
Biuret reagent
15
43
Brodies fluid
16
Hydrogen gas
44
Bromine water
17
Hydrogen peroxide
45
Buffer solutions
18
Indicators (acid-base)
46
19
Indicator (universal)
47
20
48
Carbon dioxide
21
49
Cerium(IV) solutions
22
Iodine solution
50
Chemiluminesence reactions
23
Iron(II) solutions
51
Chlorine gas
24
Iron(III) solutions
52
Chlorine water
25
Lead(II) nitrate(V)
53
26
Lithium chloride
54
27
Magnesium sulfate(VI)
55
Manganese(II) sulfate(VI)
56
Citric acid
28
Mercury solutions
57
Clock reactions
29
Methanal solution
58
Methanoic acid
59
Recipe sheet
Number
Recipe sheet
Number
Nickel sulfate(VI)
60
Sodium hydrogencarbonate
84
Nitric(V) acid
61
Sodium hydroxide
85
62
86
Oscillating reactions
63
Oxygen gas
64
Phosphoric(V) acid
65
Sodium thiosulfate
87
66
88
89
67
90
91
Potassium chloride
68
92
Potassium chromate(VI)
69
93
Potassium dichromate(VI)
70
94
Potassium hydroxide
71
Strontium chloride
95
Potassium iodide
72
Sulfur dioxide
96
Potassium manganate(VII)
73
97
74
Sulfuric(VI) acid
98
99
75
100
Sandells solution
76
101
Silver nitrate(V)
77
102
Slime
78
Tin(II) chloride
103
79
104
Sodium carbonate
80
105
81
Sodium chloride
82
Zinc sulfate(VI)
106
Sodium ethanoate
83
Agar
For microbiological activities using purchased media, follow the instructions on the bottle.
The recipes are grouped into agars for microbiology and agars for other activities. All agars for
microbiological work need to be sterilised before and after use.
General Hazards
Control
measures
Procedure for
preparing
technical agar
(also called
agar-agar)
Usual sterilising
conditions
Agar inhaled as a fine powder may cause an allergic reaction or other respiratory
problems. The use of agar that could isolate human pathogens (eg, blood agar) should
be avoided.
Use a balance in a non-working fume cupboard, ie, not switched on, with the sash
down, to weigh out agar. Use heatproof gloves to protect from scalding when handling
freshly-sterilised molten agar.
Mix 1.5 g of agar with 10 ml of water into a paste. Slowly add more water with stirring
until the volume is 100 ml. Heat the mixture with stirring on a boiling water bath to
95 C in the required container. This preliminary heating can be omitted if the agar is
going to be sterilised immediately, unless it is necessary to decant the agar into
smaller containers. In acid media, the amount of agar should be increased from 1.5 to
2 g. If the solidified agar in any recipe is too sloppy or too firm, repeat the procedure
using slightly more or less agar.
The agar gel is not stable in strongly alkaline solutions.
(If required) Autoclave the container(s) with the made-up suspension(s) for 15 minutes
at 15 psi (121 C).
Use 3.6 g of China blue lactose agar powder in 100 ml of distilled water. If this mix
proves too thin for rough handling by students, then thicken by adding 0.5 g of agaragar (just thickener, no nutrients).
Crystal violet
agar. A selective
(against Gram
positive) medium
for soil bacteria
Glucose nutrient
agar
Malt agar for fungi
In a fume cupboard which is not switched on, add 0.005 g of crystal violet (HARMFUL,
DANGEROUS FOR THE ENVIRONMENT) to 1 litre of liquid nutrient agar solution. The
resultant solution is low hazard.
Mannitol yeast
extract agar for
growing root
nodule bacteria
Nitrogen-free
mineral salts agar
for growing
nitrogen-fixing
bacteria
Mix 2 g of Bovril, 0.5 g of sodium chloride and 1.5 g of agar with 10 ml of water into
a paste. Slowly add more water with stirring until the volume is 100 ml. Heat/sterilise
the suspension as in Usual sterilising conditions above.
Mix 3 g of light malt powder (from home-brewing shops), 0.5 g of peptone (to
promote growth) in 20 ml of water. Also make a paste containing 1 g of soluble
starch in 10 ml of hot water. Add these two solutions to 1.5 g of agar with stirring and
slowly add more water with stirring until the volume is 100 ml. Stir before decanting
into smaller containers (if required) and sterilising. Autoclave the suspension at
10 psi (115 C) for 10 minutes.
CLEAPSS 2011
Mayonnaise agar
for lipase activity
(1)
Dilute 4 g salad cream or mayonnaise with 5 ml water and add 1 ml 0.1 M sodium
hydroxide solution (IRRITANT). Add about 1 ml of this alkaline mixture to a solution of
bromocresol green dye (about 0.003 g in 100 ml water) until the mixture just turns
blue-green and stir to ensure even distribution. Boil the resultant mixture with 2 g
agar, cool to 50-60 C then pour thin layers in Petri dishes. The plates will need to be
incubated at 30 C for 24 hours before being examined for orange-yellow areas
produced by lipases.
Alternatively, mayonnaise agar can be made up without the dye and, after
incubation, the plates can be flooded with 0.4 M copper(II) sulfate solution and left for
30 minutes before being examined. Clear areas in the blue-green matrix indicate
where lipases have broken down the fatty acids in the mayonnaise.
Stir together 2 g low-fat milk powder (Marvel is recommended as it contains very little
fat), 1 g agar and 100 ml water. Heat as for technical agar and pour into Petri dishes
in very thin layers. Proteases should produce clear patches by breaking down
proteins in the milk within 30 minutes or so.
Use 2 g agar in 100 ml boiling distilled (or deionised) water in a beaker. Add 10 ml of
0.2 M sodium carbonate solution and 5 ml of phenolphthalein into the beaker and stir
well.
Carbon dioxide in the atmosphere causes the colour to fade on storage, so the agar
is better prepared shortly before it is required.
Mayonnaise agar
for lipase activity
(2)
Milk agar for
protease activity
Phenolphthalein
indicator agar
Mix a paste containing 1 g of soluble starch in 10 ml of cold water. Add 1.5 g of agar,
stir well and slowly add more water with stirring until the volume is 100 ml. Heat as
for technical agar above.
CLEAPSS 2011
% ethanol
Flash point (C)
Density (g cm-3) at 20 C
Hazard
0
1.00
-
10
49
0.98
20
36
0.97
30
40
29
26
0.96
0.95
FLAMMABLE
50
24
0.93
60
22
0.91
70
21
0.89
80
20
0.86
90
17
0.83
HIGHLY FLAMMABLE
Weigh out 116 g of propanone in a tared container or measure out 147 ml of propanone in a
measuring cylinder.
Add this to a 1000 ml measuring cylinder or 1 litre measuring jug.
Add water up to the 1 litre mark.
The solution is low hazard.
CLEAPSS 2011
100
13
0.78
Alginate beads
In studies of enzymes or the physiology of yeast cells, a valuable technique is to immobilise the enzyme or
cells inside beads of sodium alginate. The beads containing the enzyme/cells can then be used as usual or
packed into a column (eg, a syringe barrel) and a suitable substrate passed over them. The products are
collected at the bottom of the column and the immobilised enzymes or cells can be used again.
When making up the alginate and enzyme solutions it is essential to use purified water; otherwise calcium
ions in the water will cause the alginate to set prematurely.
Alginate beads can usually be stored overnight, covered and refrigerated but are unlikely to keep longer than
their non-immobilised components.
Always trial practicals to confirm activity of organisms or enzymes.
Preparing immobilised enzymes/cells in alginate beads
Make up a solution of the enzyme to be studied (see Recipe sheet 37 for enzymes), or a suspension
of yeast cells using purified water.
Sprinkle 2 g of sodium alginate in 100 ml of warm, purified water and mix using a mechanical stirrer.
Allow the solution to cool. Initially the mixture will form glutinous lumps, but it becomes smooth over
time.
Dissolve 3 g of calcium chloride-6-water in 200 ml of purified water in a 250 ml beaker.
Mix 2 ml of the enzyme/suspension with 8 ml of the 2% sodium alginate solution. Variations on these
proportions may be used.
Draw this up into a 10 ml syringe.
Add the sodium alginate/enzyme (or cell) mixture one drop at a time to the calcium chloride solution
making sure the tip of the syringe is held above the solution in the beaker.
Allow the beads to harden for a few minutes before straining them out of the beaker.
CLEAPSS 2011
Aluminium solutions
Hydrated aluminium salts such as the chloride (AlCl6.6H2O, M = 241.5 g mol-1), sulfate(VI) (Al2(SO4)3.16H2O,
M = 630 g mol-1) and nitrate(V) (Al(NO3)3.9H2O, M = 375 g mol-1) absorb water (ie, they are hygroscopic) and
become damp on storage. Do check these chemicals before use.
Aluminium potassium sulfate(VI), also known as potassium aluminium sulfate, alum and potash alum is
easily stored and suitable for all activities where aluminium ions are required for testing. However, it is not all
that soluble in water, although it does make large octagonal crystals. Aluminium solutions are acidic.
Aluminium potassium sulfate(VI)
Molar mass: 474.39 g mol-1
Formula: AlK(SO4)2.12H2O
General Hazards
1000
Hazard warning
label
47.44
94.88
114
Formula: AlCl3.6H2O
General Hazards
Hazard warning
label
IRRITANT
CLEAPSS 2011
Ammonia gas
Ammonia is less dense than air and very soluble in water so it has to be collected by the downward
displacement of air (upward delivery).
Theoretically, 2.1 ml of fresh concentrated (880) ammonia solution produces 1 litre of gas, although this is
never achieved in practice, so always use an excess. Older stocks of concentrated ammonia will be less
concentrated.
If the fountain experiment is to be carried out, a round bottom flask is substituted for the gas jar in the
diagram below. Many standard text books use a calcium oxide drying tube. There is no real need for this.
General Hazards
See Hazcards 5 & 6. Ammonia begins to bubble off from 880 ammonia at about
55 C so heat gently and use anti-bumping granules to allow the ammonia to boil
off gradually.
Anti-bumping
granules
10 ml of concentrated
ammonia solution
Heat
gently
CLEAPSS 2011
The concentration of 35% (w/v) ammonia solution, (also known as 880 ammonia), is 18.1 mol dm-3.
Educational suppliers commonly supply this concentration although other concentrated solutions, eg,
25% (w/v) are available.
If kept for long periods, the concentration of ammonia solutions decreases because of leakage of gas from
the container.
If the concentrated ammonia is several years old it would be wise to test the concentration of the solution
before diluting further.
It is sensible not to store diluted ammonia solutions for long periods. It is better to prepare diluted solutions
only when required.
Solutions less than 1 M should be made by further dilution of 1 M ammonia solution and are best made fresh
before use.
If you have other concentrations of concentrated ammonia, then contact CLEAPSS for more advice. Some of
the values below are different from previous Recipe Cards as more precise information is now available.
Molar mass: 17.03 g mol-1
Formula: NH3
General Hazards
Solubility: infinite
Hazard warning
label
IRRITANT
Use a fume cupboard if concentrated ammonia or solutions more concentrated than 5 M ammonia are
used. Wear goggles (a face shield is preferable when handling large volumes) and chemical resistant
gloves.
Measure the indicated quantity of ammonia solution in an appropriate measuring cylinder.
Add the liquid to about two thirds of the final volume of water in an appropriate beaker or laboratory
jug.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
Ammonium chloride
Molar mass: 53.5 g mol-1
Formula: NH4Cl
General Hazards
2.68
5.34
40
Hazard warning
label
-
CLEAPSS 2011
Wear goggles.
Dissolve 1.17 g of ammonium vanadate(V) in 20 ml of 2 M sodium hydroxide in a beaker. (The odour
of ammonia may be detected but it will cause no harm.)
Transfer the solution to a 100 ml measuring cylinder.
Add 1 M sulfuric(VI) acid to bring the total volume to 100 ml.
The yellow solution is irritant because of the presence of sulfuric(VI) acid.
CLEAPSS 2011
Azo dyes
It is a myth that all azo dyes are carcinogens. The dyes can be prepared in schools as long as sensible
laboratory procedures are observed. On the whole, the more soluble a dye and the presence of sulfonic acid
groups on the benzene ring, then the safer the dye (eg, Orange II and methyl orange).
See Guide G195 for preparation of azo dyes from ethyl 4-aminobenzoate.
The recipes below can be used safely in schools.
General Hazards
* Other phenols, benzene diols, cresols and naphthols can be used with the diazonium salt of sulfanilic acid to produce other dyes.
10
CLEAPSS 2011
10
Barium solutions
Purified water should be used to avoid cloudiness caused by the precipitation of barium sulfate(VI).
Barium chloride is classified as TOXIC if swallowed and barium nitrate(V) as HARMFUL if swallowed.
All solutions of barium nitrate(V) are LOW HAZARD.
Barium chloride is more soluble in water than barium nitrate(V).
General Hazards
Barium chloride; see Hazcard 10A. Barium nitrate(V); see Hazcard 11.
Barium chloride
Molar mass: 244.26 g mol-1
Formula: BaCl2.2H2O
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
0.5 M
Saturated (20 C)
Hazard warning
label
HARMFUL
HARMFUL
TOXIC
Barium nitrate
Molar mass: 261.37 g mol-1
Formula: Ba(NO3)2
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
Saturated (20 C)
2.61
9
Hazard warning
label
-
Wear eye protection. Wear disposable nitrile gloves when weighing and preparing the solution.
Measure out the indicated quantity of the solid barium salt.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
11
11
Benedicts solution or DNSA (see Recipe sheet 34) should be used in place of Fehlings solution to test for
reducing sugars because it is less hazardous.
Glucose, lactose and maltose are reducing sugars and give a positive test. Sucrose is a non-reducing sugar
and does not give a positive result.
Benedicts solution is less satisfactory in testing for non-reducing sugars and aldehydes in organic chemistry
in which case Fehlings or Sandells solutions will be needed.
No hazard warning symbol is required on the bottle as the concentrations of each of the constituents are low.
This solution is not suitable for colorimetric work. See Recipe sheet 12 for Benedicts quantitative reagent or
Recipe sheet 34 for DNSA.
To differentiate between reducing sugars, enzyme tests are required.
General Hazards
Sodium carbonate; see Hazcard 95A. Copper sulfate(VI); see Hazcard 27C.
The material under test is mixed with about 1 ml of water in a test tube or vial, and about 3 ml of
Benedict's reagent is added.
Place the test tube in a boiling water bath for about 5 minutes.
The colour should progress from blue (with no glucose present) to green, yellow, orange, red, and
then brick red or brown as glucose concentration increases.
12
CLEAPSS 2011
12
The procedure detects the loss of blue colour as the sugar reduces the copper(II) ions to copper(I).
It can be used either in volumetric or colorimetric methods.
The addition of thiocyanate forms a complex and prevents the precipitation of copper(I) oxide.
See also Recipe sheet 34 for DNSA which is an alternative test for reducing sugars.
General Hazards
Sodium carbonate; see Hazcard 95A. Copper sulfate(VI); see Hazcard 27C.
Potassium thiocyanate; Hazcard 95C. Potassium hexacyanoferrate(II) see
Hazcard 79. The volumetric method uses hot liquids.
CLEAPSS 2011
13
13
Test for
soluble
proteins
Marquiss
reagent
Test for
alkaloids
Ninhydrin
For amine
groups
Sakaguchi
test
Test for
proteins
containing
arginine
See Hazcards
62, 93, 98A.
Solution A and
the combined
solution are
TOXIC.
See Hazcards
63 and 98.
Label the
solution
CORROSIVE and
TOXIC.
See Hazcard
66. No hazard
warning is
required on the
solution.
See Hazcards
70, 89 & 91.
Carbohydrate tests
See also 3,5-dinitrosalacylic acid (DNSA), Benedicts solution, Sandells solution and Fehlings solution.
For all
Wear goggles. Dissolve 5 g of napthalen-1-ol in 100 ml of
See Hazcards
Molischs
carboethanol.
40A, 70 & 98.
solution
hydrates
Label the
The solution containing a possible carbohydrate is
combined with a small amount of Molisch's reagent in a test solution HIGHLY
FLAMMABLE &
tube. After mixing, a small amount of concentrated
HARMFUL
.
sulfuric(VI) acid is slowly added down the sides of the
sloping test tube, without mixing, to form a lower layer. Look
for a purple ring at the interface of the two layers.
For
Wear goggles and chemical-resistant gloves. Dissolve 1 g
Iodic(VII) acid is
Periodic
polyCORROSIVE &
of iodic(VII) acid (periodic acid) in 100 ml of water. Used in
acid Schiff
saccharides
OXIDISING. No
conjunction with Schiffs reagent. Changes from colourless
(PAS)
hazard warning
to purple.
reaction
is required on
the solution.
14
CLEAPSS 2011
Cellular respiration
Janus
green B
Methylene
blue
TTC
Wear eye protection and disposable nitrile gloves when making up the
solution. Dissolve 0.3 g of the dye in 100 ml of purified water. Dilute this
solution ten times with water before use. Colour changes are from blue to
salmon pink.
See Guidance leaflet PS88 for more details including a practical activity.
Wear eye protection, and gloves to avoid staining the skin.
Dissolve 1 g solid in 100 ml water and add 0.6 g sodium chloride. The blue
indicator turns colourless as the dye is reduced.
No hazard
label is
required on
the solution.
See Hazcards
32 and 40. No
hazard label is
required on
the solution.
Low hazard.
The solid is
VERY TOXIC;
see Hazcard
35. The
solution is low
hazard.
Vitamin C
DCPIP
solution
CLEAPSS 2011
15
14
The solutions need to be kept acidic to avoid the formation of insoluble basic salts.
Molar mass: 395 g mol-1
Formula: Bi(NO3)3.5H2O
General Hazards
16
CLEAPSS 2011
15
Biuret reagent
Sodium hydroxide (solid) and 2 M solution. See Hazcards 91. Copper sulphate, see
Hazcard 27C.
Wear goggles.
Weigh out 0.75 g of copper(II) sulfate(VI)-5 -water.
Prepare 1 litre of 2 M potassium or sodium hydroxide solution.
Dissolve the copper(II) sulfate(VI) in the alkali and label the solution CORROSIVE.
Sodium hydroxide (solid) and a 2 M solution see Hazcard 91. Copper(II) sulfate(VI)
see Hazcard 27C.
Wear goggles.
Dissolve 1.5 g of copper(II) sulfate(VI)-5-water crystals and 6 g of potassium sodium tartrate-4-water in
500 g of purified water.
Add 375 ml of 2 M sodium hydroxide with stirring.
If a precipitate occurs, add 1 g of potassium iodide.
Pour this mixture into a 1000 ml volumetric flask and dilute to 1 litre. Mix well. Label this solution
CORROSIVE as it is a 0.75 M sodium hydroxide solution.
For quantitative analysis, a series of standards can be produced with solutions of varying % dilutions of a
protein such as albumen, using a colorimeter with a 540 nm (green) filter. Unknown proteins solutions can
then be compared against these standards.
CLEAPSS 2011
17
16
Brodies fluid
A.
Sodium azide is VERY TOXIC and contact with acids liberates a very toxic gas. (See
Hazcard 95B). For eosin, see Hazcard 32. Once in solution, the solution is LOW
HAZARD. Sodium azide may be omitted but the fluid will not keep as well and there
will be mould growth.
B.
C.
44 g of sodium bromide.
1 g of liquid detergent (eg washing-up liquid).
0.3 g of Evans blue.
Simple version for short-term use
Use water with a food dye such as cochineal and a few drops of detergent.
18
CLEAPSS 2011
17
Bromine water
The solubility of bromine in water is 4 g, (ie, 1.25 cm3 ) in 100 g of water at room temperature. This would be
a 4% (w/v) solution. And its concentration would be 0.25 M.
Solutions equal to or greater than 0.06 mol dm-3 (ie, a 0.3% v/v solution) are TOXIC.
Solutions equal to or more concentrated than 0.006 mol dm-3 (0.1% w/v or 0.03% v/v) but more dilute than
0.06 mol dm-3 should be labelled HARMFUL.
Aqueous solutions of bromine should be prepared just before use. If stored for long periods, especially
though the summer, the solution becomes paler as bromine vapour is lost.
Do not make this solution for the first time without seeking practical advice from a more-experienced
colleague.
General Hazards
Bromine is VERY TOXIC and CORROSIVE (see Hazcard 15A). Hazcard 15B deals with
bromine water. 0.025 M bromine water has a considerable vapour of bromine gas
above it. It should be dispensed from a fume cupboard. More-dilute solutions can
be used in a well-ventilated room but staff should discourage any direct inhalation
of the vapour.
Sodium chlorate(I) is CORROSIVE (see Hazcard 89), 2 M hydrochloric acid; see
Hazcard 47A, Potassium bromate(V) is TOXIC, (see Hazcard 80).
CLEAPSS 2011
19
18
Buffer solutions
Buffer solutions retain their pH on addition of small amounts of acid, alkali or on dilution.
Unless stated otherwise, buffers are low hazard but eye protection should be worn during preparation.
The pH value of a buffer is slightly altered by temperature but this is not usually significant. The values used
in the Recipe Sheet are for 20 C.
A definition for pH suitable for schools is minus the logarithm (to the base 10, ie log10) of the hydrogen ion
concentration in an aqueous solution. For a more advanced explanation, consult
http://en.wikipedia.org/wiki/PH.
There is also a pOH scale and pH + pOH =14.
A pH meter should be calibrated with standard buffers before it is used. These may be prepared from tablets
or commercial solutions.
Use distilled or deionised water to make up buffers.
Single-component buffers are very quick to make up but the values may not always be convenient.
The majority of buffers involve two components mixed in certain proportions. For accurate work, it is wise to
check their pH with a calibrated pH meter.
A universal buffer mixture can be used to obtain an array of buffer solutions from pH 2 to 11. The stock
solution can, usefully, be stored for several months.
Buffers (especially those between pH 4 and pH 7) do not keep well. Moulds develop which affect pH
readings and block the junctions on pH probes.
Commercial buffer solutions contain a mould inhibitor which allows longer storage.
To save time buffers can be stored as pre-weighed dry components or as frozen solutions. Before use, make
sure frozen solutions are returned to room temperature, that all components are fully dissolved, and check
the pH.
Special biological buffers (eg, TRIS) are available that do not use phosphate(V) or ethanoate ions which
might interfere with some biochemical processes. They are named Good buffers after their developer,
Norman Good. Different enzymes are inhibited by different reagents, so check the protocol and choose the
correct buffer system.
pH 7 buffer
pH 9.2
buffer
pH 12.6
buffer
Low hazard
See Hazcard 36A.
Low hazard
See Hazcard 13B.
Low hazard
See Hazcard 9B.
Low hazard
See Hazcard 14.
Low hazard but wear
eye protection.
See Hazcard 18.
5
4.01
9.39
10
4
9.33
15
4
9.27
20
4
9.23
25
4.01
9.18
20
30
4.01
9.14
40
4.03
9.07
50
4.06
9.02
60
4.09
8.97
70
4.12
8.93
80
4.16
8.89
90
4.2
8.85
CLEAPSS 2011
pH7 buffer
pH10 buffer
NB
See Hazcards 14
and 91.
Although masses are given to 3 decimal places, it would be acceptable to work to 2 decimal places.
3
4
5
6
7
8
9
10
11
CLEAPSS 2011
21
General
Hazards
A (ml)
B (ml)
pH
A (ml)
B (ml)
pH
A (ml)
B (ml)
2.2
2.4
2.6
2.8
3.0
3.2
3.4
3.6
3.8
4.0
2.00
6.20
10.90
15.85
20.55
24.70
28.50
32.20
35.50
38.55
98.00
93.80
89.10
84.15
79.45
75.30
71.50
67.80
64.50
61.45
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
41.40
44.10
46.75
49.30
51.50
53.60
55.75
58.00
60.45
63.15
58.60
55.90
53.25
50.70
48.50
46.40
44.25
42.00
39.55
36.85
6.2
6.4
6.6
6.8
7.0
7.2
7.4
7.6
7.8
8.0
66.10
69.25
72.75
77.25
82.35
86.95
90.85
93.65
95.75
97.25
33.90
30.75
27.25
22.75
17.65
13.05
9.15
6.35
4.25
2.75
pH
9.6
9.8
10.0
10.2
10.4
10.6
10.8
11.0
A (ml)
5.00
6.20
10.70
13.80
16.50
19.10
21.20
22.70
pH
6.0
6.5
7.0
7.5
8.0
22
A (ml)
87.7
68.5
39.0
16.0
5.3
B (ml)
12.3
31.5
61.0
84.0
94.7
CLEAPSS 2011
TRIS buffers
pH
7.0
7.5
8.0
8.5
9.0
Volume of 0.1 M
TRIS solution
100
100
100
100
100
Volume of 0.1 M
hydrochloric acid
93.2
80.6
58.4
29.4
11.4
CLEAPSS 2011
23
19
Hydrated calcium chloride and nitrate(V) absorb water from the atmosphere. On occasions, the solid
completely dissolves to leave a clear solution.
Distilled or deionised water should be used to make solutions. In hard water areas, solutions may be cloudy
if tap water is used.
Do not use anhydrous calcium chloride to make solutions.
General Hazards
Formula: CaCl2.6H2O
Formula: Ca(NO3)2.4H2O
24
CLEAPSS 2011
20
Saturated calcium hydroxide solution, commonly called lime water, is a 0.02 M aqueous solution of calcium
hydroxide with a pH of 12.4.
It does not keep for long periods in large bottles because it reacts with carbon dioxide from the atmosphere.
It would be wise to start afresh each year.
Lime water, in small bottles designed for class use, will quickly cease to work. Class sets need to be tested
before handing out to the class.
1 M hydrochloric acid can be used to clean bottles that previously contained limewater.
Molar mass: 74.09 g mol-1
Formula: Ca(OH)2
General Hazards
Calcium hydroxide solid; see Hazcard 18. Splashed droplets of limewater in the eye
have caused quite severe irritation, so the solution should also be labelled and treated
as an IRRITANT even though strictly its dilution is such that it is not formally classed as
hazardous.
If a large stock is required, keep an excess of calcium hydroxide in an aspirator protected by a sodalime tube, as shown below, and top up with distilled water as necessary.
Use about 100 g calcium hydroxide for a 10 litre aspirator. It might take a week to fully settle.
Absorption tube
filled with soda lime
Lime
water
Calcium
hydroxide
CLEAPSS 2011
25
21 Carbon dioxide
Carbon dioxide is prepared by the action of dilute hydrochloric or nitric acid on calcium carbonate, usually as
marble chips. If powdered calcium carbonate is used, the rate of gas production may be too rapid to be
easily controlled.
The gas is collected over water or by downward delivery. (Downward delivery uses a tube into the bottom of
an upright gas jar. It is also known as upward displacement of air and relies on the fact that carbon dioxide is
more dense than air.) Although carbon dioxide is slightly soluble in water (at room temperature, the solubility
of carbon dioxide is about 6.4 cm3 of carbon dioxide per 100 ml of water), the rate at which it dissolves is
slow. However, if necessary, the gas can be collected over warm water in which it is less soluble.
Collection by downward delivery is quicker but it is difficult to ascertain exactly when the gas jar is full.
However, downward delivery is necessary for burning magnesium in carbon dioxide, in order to avoid water
interfering with the reaction.
Soda water is a saturated solution of carbon dioxide in water (carbonic acid).
General Hazards
Warm water
26
CLEAPSS 2011
22
Cerium(IV) solutions
Cerium(IV) compounds have a variable water content, which makes preparing accurate concentration
solutions very difficult. This is why the mass given in the recipe is approximate.
Cerium(IV) solutions, which have an intense yellow colour, are used in redox titrations.
Cerium(IV) solutions are more stable in solution than potassium manganate(VII) solutions.
Cerium(IV) solutions need to be prepared in dilute sulfuric(VI) acid.
[Hydrochloric acid must be avoided as cerium(IV) ions oxidise the chloride ions slowly to chlorine.]
Cerium(IV) solutions should be standardised against sodium ethanedioate solution before use.
General Hazards
The solids are irritating to the eyes, respiratory system and skin.
CLEAPSS 2011
27
23
Chemiluminesence reactions
See Hazcards
4B, 89, 91.
Wear goggles
to prepare the
solution.
See Hazcards
4B, 50, 79, 91.
Wear goggles
to prepare the
solution.
See Hazcards
4B, 27C, 50,
95A.
See Hazcards
4B, 9A, 25,
27C, 50, 89, 93,
95A.
28
CLEAPSS 2011
24
Chlorine gas
See Hazcards 22A, 20 & 81. Double check that you are using concentrated
hydrochloric acid and not concentrated sulfuric(VI) acid. Explosions have resulted from
using the wrong acid.
Bchner flask
At least 5 g of
Potassium
Manganate(VII)
Gas jar
Wooden block
Delivery tube as far
down as possible
CLEAPSS 2011
29
See Hazcards 22A, 20 & 89. Sodium chlorate(I) is a solution provided by the
supplier often as sodium hypochlorite. Sodium chlorate(V) is a solid. Do not get
them mixed up.
Separating funnel
Rubber tubing
5 M hydrochloric acid
Sodium
chlorate(i)
solution
Mineral wool
Magnetic stirrer
Gas jar
Delivery tube as far
down as possible
30
CLEAPSS 2011
25
Chlorine water
The solubility of chlorine in water is about 0.6 g in 100 ml of water at room temperature. This would be a
0.6% (w/v) solution. Its concentration would be 0.085 M.
An aqueous solution of chlorine should be prepared just before use. It does not keep for more than just a few
days and should not be stored.
It is very difficult to make up solutions of known concentration. All that is required for displacement reactions
is that the solution works. It should, therefore, be trialled before use.
Do not make this solution for the first time without seeking practical advice from a more experienced
colleague.
Three methods are described below.
General Hazards
Chlorine gas is TOXIC and CORROSIVE (see Hazcard 22A). Hazcard 22B deals with
chlorine water. Freshly-made chlorine water is formally LOW HAZARD but it has a
considerable vapour of chlorine gas above it. It is better dispensed from a fume
cupboard. More dilute solutions can be used in a well-ventilated room but staff
should discourage direct inhalation of the vapour.
Method 1
Use a fume cupboard. Wear goggles or a face shield and chemical-resistant gloves.
Bubble chlorine gas into 250 ml water in a gas jar until the solution goes light green. Use the
equipment for preparing chlorine gas but fill the gas jar half-full with water.
Method 2
Sodium chlorate(I) solution is an aqueous solution. Do not become confused with sodium chlorate(V), which
is a solid.
10% w/v available chlorine sodium chlorate(I) solution does not store well. Over two years it may become
completely useless.
Use a fume cupboard. Wear goggles or a face shield and chemical-resistant gloves.
Place 10 ml of 10% w/v available chlorine sodium chlorate(I) solution (CORROSIVE) in 1 litre beaker.
Add about 80 ml of water and 10 ml of 2 M hydrochloric acid. Stir well.
Dilute to a suitable volume with more water.
Method 3
Sodium dichloroisocyanurate is used for purifying water.
Use a fume cupboard. Wear eye protection.
3
Add 3 g of sodium dichloroisocyanurate (OXIDISING; HARMFUL), to 100 cm of water. When the solution
3
-3
is clear, add 100 cm of 1 mol dm hydrochloric acid.
CLEAPSS 2011
31
26
A mixture of compounds is placed on a stationary phase such as paper, or silica on thin layer plates.
Chromatography involves passing a solvent through the stationary phase which separates the components
of the mixture due to subtle differences in a compound's partition coefficient between the mobile solvent and
the stationary phase.
A locating agent is used to emphasise where the components of the mixture end up after the
chromatography has finished. This is particularly important for colourless components of mixtures.
Control measures
for all of the
following solutions
Substances
separated
Amino acids
Analgesics,
eg, aspirin,
paracetamol
Anthocyanins
(plant
pigments)
Inks from
Biro pens
Chlorophyll
Lipstick
Metal ions
Nitration of
methyl
benzoate
Sugars
32
HIGHLY FLAMMABLE
CLEAPSS 2011
Formula: CrCl3.6H2O
Formula: CrK(SO4)2.12H2O
CLEAPSS 2011
33
28
Citric acid
Formula: C6H8O7
Formula: C6H8O7.H2O
Hazard warning
label
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of the solid citric acid.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
34
CLEAPSS 2011
29
Clock reactions
The thiosulfate/acid
reaction
(disappearing
cross)
The potassium
iodate/sodium
metabisulfite
reaction
(The Landolt iodine
clock)
The reaction
between iron(III)
ions and iodide
ions
The potassium
iodide/potassium
persulfate reaction
CLEAPSS 2011
35
30
General Hazards
Cobalt salts are TOXIC (See Hazcard 25). Reported carcinogenic and sensitisation
warnings about cobalt compounds have become more severe in recent years.
CLEAPSS has not received any reports from educational establishments of cobalt
salts causing harm. The concerns for health are in mining and metallurgical
applications used on a daily basis and in large doses.
Formula: CoCl2.6H2O
Wear eye protection. Wear disposable gloves and consider weighing the solid in non-working fume
cupboard with the sash window low enough to stop any particles being inhaled.
Dissolve 2.38 g of cobalt(II) chloride-6-water in 70 ml of water.
Make up to 100 ml with pure water.
The solution should be labelled TOXIC.
Formula: Co(NO3)2.6H2O
Use the method above but dissolve 2.91 g of cobalt(II) nitrate-6-water in 70 ml of water.
Formula: CoSO4.7H2O
Use the method above but dissolve 2.81 g of cobalt(II) sulfate(VI)-7-water in 70 ml of water.
Thermochromic liquid
Used in models of hot-water systems.
36
CLEAPSS 2011
31
Copper(II) solutions
Formula: CuSO4.5H2O
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
0.5 M
1.0 M
Saturated (20 C)
Formula: CuCl2.2H2O
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
0.5 M
1.0 M
Saturated (20 C)
Formula: Cu(NO3)2.3H2O
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
0.5 M
1.0 M
Saturated (20 C)
Procedure
CLEAPSS 2011
37
32
Real crude oil (and petrol) contain benzene in concentrations greater than 0.1% and must not be used in
school science (COSHH Regulations).
A synthetic mixture can be prepared using mainly aliphatic hydrocarbons to illustrate the principle of
fractional distillation of crude oil in industry.
An alternative name for petroleum spirit is petroleum ether.
On storing for several months, the lighter fractions evaporate away so the procedure should be tested to see
that enough low boiling point fractions can be distilled off.
Further details on the procedure can be found in L195, Safer Chemicals, Safer Reactions and section 13 of
the Handbook.
Standard fractional distillation equipment using a fractionating column is unsuitable as very high
temperatures are required. Use apparatus similar to the diagram below.
General Hazards
Wear eye protection. Do not prepare the mixture near sources of ignition.
Mix together 55 ml of liquid paraffin (medicinal), 20 ml of paraffin oil (kerosene), 11 ml of white spirit,
4 ml of petroleum ether (100-120 C), 4 ml of petroleum ether (80-100 C) and 6 ml of petroleum ether
(60-80 C).
Add a squeeze of black oil paint (eg, Winsor and Newtons Ivory Black) from a tube and stir well.
After adding to a labelled bottle, shake the mixture well. Label the container HIGHLY FLAMMABLE &
HARMFUL.
Always shake the mixture well before use. The paint will separate out if the bottle is left still for some
time.
Note
The recipe is not written in stone. Another possibility, if you have an even lower fraction of petroleum spirit,
is:
Mix together 50 ml of liquid paraffin (medicinal), 20 ml of paraffin oil (kerosene), 10 ml of white spirit,
5 ml of petroleum ether (100-120 C), 5 ml of petroleum ether (80-100 C), 5 ml of petroleum spirit
(60-80 C) and 5 ml of petroleum spirit (40-60 C).
0 360 C thermometer
75 x 10 mm test tubes
to collect the fractions
1 ml of water in a
test tube to compare volumes
(optional)
Block of wood to
hold the test tubes
Heat
38
CLEAPSS 2011
33
2,4-Dinitrophenylhydrazine solution
The solid is also known as 2,4-DNP and the solution is called Bradys reagent.
It is used to identify organic compounds with carbonyl groups by producing orange or yellow insoluble
derivatives which, if purified by recrystallisation, give sharp melting points.
Solid 2,4-DNP is supplied moist as there may be an explosion risk if very dry solid is handled. Unopened
bottles of 2,4-DNP already contain 20% to 33% water.
Two recipes for preparing Bradys reagent are given below. The new recipe using phosphoric acid has been
shown to work and is safer.
General Hazards
Formula: (NO2)2C6H3NHNH2
Procedure for making about 100 ml of Bradys reagent
Traditional method
New method
Wear eye protection and gloves.
Wear eye protection, gloves and work in a fume
cupboard.
Stir 2 g of 2,4-DNP with 50 ml of 85%
Stir 2.7 g of 2,4-DNP with 96 ml of methanol.
phosphoric(V) acid.
Cautiously and slowly add 4 ml of concentrated
Add 50 ml of ethanol.
sulfuric(VI) acid.
Usually the solid dissolves to form a clear
solution with a little stirring. If cloudy, filter.
Label the solution TOXIC and HIGHLY FLAMMABLE.
CLEAPSS 2011
39
34
3,5-Dinitrosalicylic acid
3,5-Dinitrosalicylic acid is HARMFUL by ingestion and irritating to the eyes and skin.
Wear goggles.
Add 1.0 g of 3,5-dinitrosalicylic acid to 50 ml of water and warm gently to dissolve.
Slowly add 30 g sodium potassium tartrate tetrahydrate, (KNaC4H4O64H2O). The mixture thickens
(like custard).
Add 20 ml of 2 M sodium hydroxide solution and dilute the mixture with water to a final volume of
100 ml.
Label the solution IRRITANT.
Experimental procedure
40
CLEAPSS 2011
35
The food-base should pour easily when hot but set firmly when cold. It should also remain firm when the
adult flies emerge.
Preparing the food base
CLEAPSS 2011
41
36 Electroplating solutions
There are many formulations for plating solutions. Some are hazardous and where they are not covered by a
model (general) risk assessment, either here or elsewhere, a special risk assessment will be needed before
use by teachers / technicians or students. Help can be obtained from CLEAPSS.
Use
Copper
electroplating
Nickel
electroplating
Silver
electroplating
Zinc electroplating
42
Notes
See Hazcard 27C. Label the
solution HARMFUL.
See Hazcard 65B. Label the
solution HARMFUL.
See Hazcards 87 and 98A. Label
the solution IRRITANT.
CLEAPSS 2011
37
Enzymes
See Hazcard 33. All enzyme powders are HARMFUL and may cause sensitisation.
Any solutions greater than 1% should be considered as IRRITANT.
Wear eye protection and disposable nitrile gloves when making up solutions from the powders.
Avoid inhaling the powder. It may be wise to carry out the weighing and preparation in a fume
cupboard (with the fan switched off to avoid the draught affecting the balance).
Measure out 1 g of the enzyme.
Add the solid to about 70 ml of water or buffer (if needed for the activity) in a beaker.
Stir to dissolve (do not warm the solution).
Pour the solution into an appropriate measuring cylinder/volumetric flask.
Dilute to the final volume with pure water.
Pour into a labelled bottle and mix well. Store in the refrigerator or on ice during use.
Carry out the procedure that the students will undertake and consult with the teacher to confirm that
the results are satisfactory. It may be necessary either to dilute the solution further with more water or
add more of the enzyme.
See Hazcard 95C. Although low hazard, bile salts have an unpleasant odour and
may be better prepared in the fume cupboard.
CLEAPSS 2011
43
38
Etching solutions
There are many formulations for etchants. Some are hazardous and where they are not covered by a model
(general) risk assessment, here or elsewhere, a special risk assessment will be needed before use by
teachers / technicians or students. Help can be obtained from CLEAPSS.
Use
Aluminium
etching
Copper etching
Iron etching 1
(Frys reagent)
Iron etching 2
(Nital solution)
Lead etching
Notes
See Hazcard 91, Wear goggles.
44
CLEAPSS 2011
39
Ethanoic acid
Formula: CH3COOH
General Hazards
See Hazcard 38A. Ethanoic acid is CORROSIVE. It has a very sharp odour.
500
14
29
57
2500
71
143
286
Hazard warning
label
IRRITANT
Procedure
Wear goggles (a face shield is preferable for large volumes) and chemical resistant gloves.
Use the fume cupboard.
Measure out the indicated volume of ethanoic acid in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir the mixture well.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
45
40
Fehlings solutions
Benedicts solution or DNSA are safer alternatives when testing for sugars.
Fehling's solution can only be used to test for aliphatic aldehydes, whereas Tollen's reagent (see Recipe
Sheet 102) can be used to test for both aliphatic and aromatic aldehydes.
Fehling's solutions A & B need to be made and stored separately. They are mixed just prior to use.
Sandells solution (which is little known and not cited in exam school texts; see Recipe Sheet 76) can be
made and stored for long periods. The concentration of sodium hydroxide is only 0.4 M in the solution
(Fehlings solution B is close to 4M).
General Hazards
Fehlings solution A
Fehlings solution B
Wear goggles.
Measure out 15.4 g of sodium hydroxide and 35 g of potassium sodium tartrate.
Add the solids to 60 ml of water on a magnetic stirrer. Make up to 100 ml with water. Label the bottle
and add a CORROSIVE warning.
Solutions A and B should be mixed in equal volumes, just prior to use. During the test, do not heat directly
over a Bunsen burner: use a boiling water bath.
46
CLEAPSS 2011
41
Formalin-alcohol
fixative for plant
tissue
Formalin-acetoalcohol (FAA)
fixative for plant
tissue
Acetic alcohol
(Clarkes fluid) (a
cytological fixative
especially for
chromosomes)
CLEAPSS 2011
47
42
Carbon
monoxide
Dinitrogen
monoxide
Hazard
information
See Hazcard 21
See Hazcard 68
Hydrogen
chloride
See Hazcard 49
Hydrogen
sulfide
Nitrogen
See Hazcard 51
Nitrogen
dioxide
See Hazcard 68
Nitrogen
monoxide
See Hazcard 68
See Hazcard 93
48
CLEAPSS 2011
43
Hydrochloric acid
Hydrochloric acid is available from all major educational suppliers as 35 - 38% (w/w) solution with a specific
gravity of 1.18. If you wish to know the dilution factors for other commercial concentrations (eg, 32%), then
contact CLEAPSS.
For accurate work, the concentration should always be found by titrating against a standard solution of
sodium carbonate.
The concentration of concentrated hydrochloric aid is about 11.7 mol dm-3.
If kept for long periods, the concentration of 35 - 38% (w/w) hydrochloric acid decreases as the gas diffuses
into the atmosphere.
Do not make dilute solutions for the first time without seeking practical advice from a more experienced
colleague.
Density: 1.18 g cm-3
Formula: HCl
General Hazards
Concentration
required
0.01 M
0.1 M
0.4 M
0.5 M
1M
2M
See Hazcard 47A. Hydrochloric acid is CORROSIVE. It has a very sharp odour.
500
17
21
42
84
2500
84
105
209
417
Hazard warning
label
IRRITANT
Procedure
Wear goggles (a face shield is preferable when handling large volumes) and chemical resistant
gloves.
Use a fume cupboard. Take care opening a bottle on a hot day.
Measure out the indicated quantity of concentrated hydrochloric acid in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir the mixture well.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
49
44 Hydrogen gas
General Hazards
See Hazcards 27C, 48 & 98. Hydrogen is extremely flammable. There must be no
Bunsen burner being used within 1 m of the apparatus.
Note that the first gas collected consists of air and hydrogen. This is an explosive
mixture and should be discarded.
Wear goggles.
Place several granules of zinc in the
generator.
The reaction needs to be speeded up at the
start, to flush out air quickly.
Add about 5 ml of 1 M copper sulfate(VI)
(HARMFUL) down the thistle funnel. This reacts
with zinc, coating it with copper which acts as
a catalyst.
Add 2 M sulfuric(VI) acid (CORROSIVE) down
the thistle funnel.
Collect the gas in the 250 ml measuring
cylinder. Once the measuring cylinder is full,
its contents (largely air) must be discarded.
Now gas jars, boiling tubes, syringes and soap
bubbles of pure hydrogen can be prepared.
50
1 to 5 ml of 1M
copper(II) sulfate(VI) plus
2M sulfuric acid
250 ml measuring
cylinder
250 ml
plastic bottle
Hydrogen
generator
Zinc
500ml measuring
cylinder;
to ensure
all air is flushed
through
U-tube
Calcium chloride
CLEAPSS 2011
45
Hydrogen peroxide
In the UK, the concentration of hydrogen peroxide is given in terms of the volume of oxygen it can produce;
thus 1 ml of 10 vol hydrogen peroxide is capable of producing 10 ml of oxygen gas at 0 C and 1atm.
The maximum concentration of purchased hydrogen peroxide is 100 vol and even this solution will become
more dilute with time.
The concentration of 100 vol solution is 8.3 mol dm-3.
Commercial hydrogen peroxide solutions contain an inhibitor to slow down decomposition. During the
preparation of diluted solutions, the inhibitor is diluted so solutions will deteriorate quite quickly. They should
be prepared when required and not stored for long periods.
For accurate work you will need to standardise the solution against a known solution of potassium
manganate(VII).
For activities with catalase, start with a 1 M solution and adjust subsequent concentrations depending on
observed results.
Molar mass: 34.02 g mol-1
Formula: H2O2
General Hazards
Concentration
required
0.08 M
0.1 M
0.34 M
1M
1.7 M
3.4 M
1 vol
1.2 vol
4 vol
12 vol
20 vol
40 vol
0.29%
0.34%
1.14%
3.3%
5.7%
11.4%
250
Hazard
warning
label
IRRITANT
IRRITANT
Procedure
Wear eye protection and gloves.
Measure out the indicated quantity of 100 vol hydrogen peroxide in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a beaker or laboratory jug and stir.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
51
46
Indicators (acid-base)
General Hazards
See Hazcard 32. The hazards of many dyes and indicators are not well known.
Many are made up in ethanol which is HIGHLY FLAMMABLE & HARMFUL (see
Hazcard 40A).
Indicator
Quantity
(g)
Volume of
IDA
Acid
Alkali
pH range
Bromophenol blue
Methyl orange
Bromocresol green
Methyl red
Bromothymol blue
Phenol red
Neutral Red
Cresol red
Thymol blue
Phenolphthalein
Thymolphthalein
0.4
0.4
0.4
0.2
0.4
0.4
0.4
0.2
0.4
1.0
2.0
200
200
200
300
200
200
200
200
200
600
1000
Yellow
Red
Yellow
Red
Yellow
Yellow
Red
Yellow
Yellow
Colourless
Colourless
Blue
Yellow
Blue
Yellow
Blue
Red
Yellow
Purple
Violet
Mauve
Blue
3.0 - 4.5
3.0 - 4.5
4.0 - 5.4
4.0 - 6.0
6.0 - 7.5
6.6 - 8.0
6.8 - 8.0
7.2 - 8.8
8.0 - 9.6
8.0 - 10.0
9.3 - 10.5
Litmus solution
Dissolve 1 g of litmus in 1000 ml of water. Other recipes suggest up to 10 g of litmus but this may
depend upon the age of the sample. Filter if necessary.
52
CLEAPSS 2011
47
Indicator (universal)
General Hazards
See Hazcard 32. The hazards of many dyes and indicators are not well known.
Ethanol is HIGHLY FLAMMABLE & HARMFUL (see Hazcard 40A).
3.5
CLEAPSS 2011
4.5
5.5
6.5
7.5
53
8.5
9.5
10
10.5 11
48
General Hazards
See Hazcard 32. The hazards of many dyes and indicators are not well known.
Many are made up in ethanol which is HIGHLY FLAMMABLE & HARMFUL (see
Hazcard 40A).
For use:
Dilute the stock solution ten times with freshly-boiled distilled water.
Bubble air through the diluted solution to equilibrate it with atmospheric carbon dioxide.
When ready for use, the solution should be a deep cherry red colour in a bottle (in a test tube the
colour of the solution is less intense). If difficulties are experienced (eg, if distilled water is too acidic),
try adding a pinch more sodium hydrogencarbonate. Avoid breathing over open vessels of the diluted
indicator; the carbon dioxide exhaled may alter its pH.
54
CLEAPSS 2011
49
Cerium(IV) titrations
Dissolve 1.49 g of 1:10-phenanthroline in 100 ml of water and add
0.70 g of iron(II) sulfate(VI). (Ferroin indicator).
Dichromate(VI) titrations
Dissolve 0.1 g of N-phenylanthranilic acid in 5 ml of
0.1 M sodium hydroxide. Dilute to 100 ml with water.
EDTA titrations
Dissolve 1 g of the Eriochrome black T (Solochrome black) in
100 ml of ethanol or grind 1 g of the solid dye with 100 g of
sodium chloride. Use about 0.2 g of this solid mixture for each
titration.
CLEAPSS 2011
55
50
Iodine solution
Formula: I2
General Hazards
See Hazcard 54. The organic solvents used are also hazardous and the relevant
Hazcard should be consulted.
1000
Hazard
warning
label
-
Procedure
Wear eye protection and work in a well-ventilated room. Wear disposable nitrile gloves.
Measure out the indicated quantity of potassium iodide (KI) into an appropriate beaker.
Moisten the potassium iodide with a few drops of water.
Measure out the indicated quantity of iodine and add it to the moistened potassium iodide.
Add a small volume of water and stir. When no more iodine appears to dissolve, add some more water
and stir. Keep repeating this procedure until all the iodine has dissolved.
Pour the solution into a measuring cylinder and dilute to the final volume. Make sure there are no bits
of iodine remaining. If there are, return the solution to the beaker and leave it on a magnetic stirrer for
several minutes.
Add the solution to a labelled bottle and mix well.
56
CLEAPSS 2011
51
Iron(II) solutions
See Hazcards 55B and 98A. Label the final solution IRRITANT because of the acid
present.
Iron(II) sulfate(VI)-7-water
Also known as ferrous sulfate heptahydrate.
Formula: FeSO4.7H2O
Molar mass: 278.01 g mol-1
Volume (ml) of solution required
Concentration
required
0.01 M
0.1 M
1M
Saturated (20 C)
100
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
Iron(II) chloride-4-water
Do not use the anhydrous salt.
Formula: FeCl2.4H2O
Concentration
required
0.01 M
0.1 M
1M
Saturated (20 C)
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of iron(II) salt.
Add the solid to about two thirds of the final volume of 1 M sulfuric(VI) acid for the sulfate(VI) or 1 M
hydrochloric acid for the chloride in a beaker or laboratory jug.
Stir to dissolve (do not warm the solution).
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
either 1 M sulfuric(VI) acid for the sulfate(VI) or 1 M hydrochloric acid for the chloride to the required
level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
57
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of diammonium iron(II) sulfate(VI).
Add the solid to about two thirds of the final volume of 1 M sulfuric(VI) acid in a beaker or laboratory
jug.
Stir to dissolve (do not warm the solution).
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
1 M sulfuric(VI) acid to the required level.
Pour into a labelled bottle and mix well. Add hazard warning.
58
CLEAPSS 2011
52
Aqueous solutions of iron(III) salts do not keep if made up solely in water. If required for a longer period, an
acid should be added. Even in acid solution, there is gradual deterioration and a solid often separates out on
storage.
Iron(III) chloride (also known as ferric chloride) is available in two forms, hydrated iron(III) chloride and
anhydrous iron(III) chloride. The anhydrous solid should not be used to make iron(III) solutions. It does not
dissolve but reacts exothermically with water, to form iron(III) hydroxide and hydrogen chloride gas.
Iron(III) chloride-6-water does not store well as it absorbs water (hygroscopic).
Iron(III) sulfate(VI) has a variable amount of water of crystallisation (9-water is just one of the varieties on
sale). It does not dissolve completely and the solution will need to be filtered (or the solid allowed to settle)
before use.
Ammonium iron(III) sulfate(VI) is much easier to use and is suitable for most applications. The lilac salt
stores quite well but the surface sometimes becomes brown/yellow if the solid has become damp. The
brown/yellow solid should be removed before weighing. The solid also deteriorates if exposed to heat and
light. The solution is brown, Solutions should not be heated to accelerate dissolving as colloidal solutions can
form.
See the etching solution on Recipe sheet 38 if the iron(III) chloride is to be used for this purpose.
Formula: FeCl3.6H2O
General Hazards
Concentration
required
0.01 M
0.1 M
1M
Saturated (20 C)
See Hazcard 55C. Do make sure you are using then hydrated salt and not the
anhydrous salt.
Volume (ml) of solution required
100
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of iron(III) chloride-6-water.
Add the solid to about two thirds of the final volume of 1 M hydrochloric acid in a beaker or laboratory
jug.
Stir to dissolve (do not warm the solution).
Pour the solution from the beaker into an appropriate measuring cylinder or a laboratory jug and add
1 M hydrochloric acid to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
59
Iron(III) sulfate(VI)
Molar mass: 562 g mol-1
Formula: Fe2(SO4)2.9.H2O
General Hazards
See Hazcards 55C and 98. 1 M sulfuric(VI) acid. Label the final solution IRRITANT
because of the acid present.
Volume (ml) of solution required
Concentration
required
0.01 M
0.1 M
0.5 M
1M
100
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
IRRITANT
Formula: NH4Fe(SO4)2.12.H2O
General Hazards
Concentration
required
0.01 M
0.1 M
0.5 M
Saturated (20 C)
See Hazcards 55C and 98. 1 M sulfuric(VI) acid. Label the final solution IRRITANT
because of the acid present.
Volume (ml) of solution required
100
250
1000
Hazard warning
label
IRRITANT
IRRITANT
IRRITANT
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of the iron(III) salt.
Add the solid to about two thirds of the final volume of 1 M sulfuric(VI) acid in a beaker or laboratory
jug.
Stir to dissolve (do not warm the solution).
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
1 M sulfuric(VI) acid to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
Procedure
The solution is painted onto paper in dull indoor light (preferably in a cupboard) and hung to dry over paper
towels or blotting paper. An opaque object (eg, scissors) is placed on the dry paper and left in the light for a
day or two although, on a sunny day, 20 minutes may well be enough for light to convert soluble iron(III) salts
to insoluble iron(II) salts.
Wearing gloves, rinse the paper in a bowl of water to remove the solution leaving a colourless image on a
Prussian Blue background. This can be messy and may stain clothing and unprotected hands. The paper is
again hung to dry over absorbent paper.
60
CLEAPSS 2011
53
Lead(II) nitrate(V)
Tap water contains sulfate and carbonate ions which will precipitate out as insoluble lead compounds if lead
ions are added. Lead salts should, thereore, be prepared in distilled or deionised water.
Lead(II) nitrate(V) is used in preference to lead(II) ethanoate (acetate) as it appears to be slightly less
hazardous.
Disposal issues: see section 7.5 of the Handbook.
Molar mass: 331.21 g mol-1
Formula: Pb(NO3)2
General Hazards
Concentration
required
0.005 M
0.01 M
0.05 M
0.1 M
1M
Saturated (20 C)
250
1000
Hazard warning
label
TOXIC
TOXIC
TOXIC
TOXIC
TOXIC
Procedure
Wear goggles and disposable nitrile gloves. If necessary, weigh materials on a balance placed in a
fume cupboard which is not switched on and the sash pulled down.
Measure out the indicated quantity of lead(II) nitrate(V).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
61
54
Lithium chloride
Molar mass: 42.39 g mol-1
Formula: LiCl
General Hazards
Lithium salts are HARMFUL if swallowed and irritants. See Hazcards 47A and 58.
250
2.12
4.24
90
1000
21.20
42.39
900
Hazard warning
label
HARMFUL
Procedure
Measure out the indicated quantity of lithium chloride.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add a hazard warning if appropriate.
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CLEAPSS 2011
55
Magnesium sulfate(VI)
Magnesium sulfate(VI)-7-water loses water of crystallisation slowly on standing to the air and becomes
powdery (efflorescent). However, it is more convenient to make solutions from the hydrated salt than the
anhydrous salt which is also available from suppliers.
The anhydrous salt readily absorbs water from the atmosphere during storage.
Molar mass: 246.47 g mol-1
Formula: MgSO4.7H2O
General Hazards
100
1000
Hazard warning
label
2.47
12.32
24.65
75
6.16
30.81
61.62
190
24.65
123.24
246.47
750
Procedure
Measure out the indicated quantity of hydrated magnesium sulfate(VI).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
CLEAPSS 2011
63
56
Manganese(II) sulfate(VI)
Manganese(II) sulfate(VI) forms many hydrates. The 4 water and 1 water varieties appear to be the most
readily available.
The values in brackets in the table below are for the monohydrate.
Molar mass: 223.07 g mol-1
Molar mass: 169.02 g mol-1
Formula: MnSO4.4H2O
Formula: MnSO4.H2O
General Hazards
250
1000
2.23 (1.69)
11.15 (8.45)
22.31 (16.90)
100 (45)
5.58 (4.23)
27.89 (21.13)
55.77 (42.26)
250 (115)
22.31 (16.90)
111.54 (84.51)
223.07 (169.02)
1000 (450)
Hazard warning
label
HARMFUL
HARMFUL
Procedure
Measure out the indicated quantity of hydrated manganese(II) sulfate(VI).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add a hazard label if appropriate.
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CLEAPSS 2011
57
Mercury solutions
Only make what is needed. Diluting to 0.04 M reduces the level of hazard to TOXIC.
The main issue with mercury salts is that they should not be put down the drain. The waste should be
collected, stored as toxic waste and removed to sealed landfill by a registered hazardous waste collector.
Although labelled toxic and having a notorious history, using an inorganic salt will not kill you. However, the
risk assessment for the activity must establish that the use of the mercury compound is really necessary and
that no alternative is possible.
General Hazards
Procedure for preparing 100 ml of 0.02 M of dimercury(I) nitrate(V)-2-water solution [which is 0.04 M
with respect to the mercury(I) ion]
Wear goggles.
Dissolve 1.12 g of the solid in 70 ml of 1 M nitric(V) acid.
Transfer the solution to a 100 ml measuring cylinder.
Make up to 100 ml with 1 M nitric(V) acid.
Label the solution CORROSIVE and TOXIC.
Mercury(II) chloride
Formula: HgCl2
CLEAPSS 2011
65
58
Methanal solution
Formula: HCHO
General Hazards
See Hazcard 63. It can cause burns, conjunctivitis or sensitisation by skin contact.
0.1 M
0.3 M
1M
1.3 M
Hazard warning
label
IRRITANT
HARMFUL
HARMFUL
HARMFUL
Procedure
Use a fume cupboard to handle the concentrated solution; wear goggles (a face shield is preferable
when handling large volumes) and wear disposable nitrile gloves.
Measure out the indicated volume of 40% methanal solution in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a suitable beaker or laboratory jug and
stir.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add the appropriate hazard warning.
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CLEAPSS 2011
59
Methanoic acid
Formula: HCOOH
General Hazards
Concentration
required
0.01 M
0.1 M
1M
2M
250
10.5
21
2500
105
210
Hazard
warning label
IRRITANT
Procedure
Wear goggles (a face shield is preferable for large volumes) and chemical-resistant gloves.
Work in a fume cupboard.
Measure out the indicated quantity of methanoic acid in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir the mixture well.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
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60
Nickel sulfate(VI)
Nickel sulfate(VI)-6-water is the most common nickel sulphate salt sold now, but do check the label on the
bottle because the 7-water hydrate is also available.
Formula: NiSO4.6H2O
General Hazards
The hazards of nickel salts are being constantly reviewed. Sensitisation may be a
problem for some people. See Hazcard 65B.
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61
Nitric(V) acid
The commercial solution is about 70% (w/v) and has a specific gravity of 1.42. Its concentration is
15.8 mol dm-3.
Concentrated nitric(V) acid should be a colourless solution and stored in dark bottles. When exposed to light,
the solution goes brown and TOXIC nitrogen dioxide gas is produced (see Hazcard 68). Nitrogen dioxide will
leak from bottles and the solution will become less concentrated.
Also available from some suppliers is 95-100% (fuming) nitric(V) acid, which is even more hazardous as it is
unstable. It is unsuitable for schools to store this for long periods. Contact CLEAPSS if you find some.
Do not make dilute solutions for the first time without seeking practical advice from a more-experienced
colleague.
Molar mass: 63.01 g mol-1
Formula: HNO3
General Hazards
Concentration
required
0.01 M
0.1 M
0.4 M
1M
5M
See Hazcard 67. If in contact with the skin, this becomes yellow and layers of skin
peel off.
Volume (ml) of solution required
500
1000
2500
Ten-fold dilution of a 0.1 M solution with water
Ten-fold dilution of a 1 M solution with water
13
25
63
32
63
158
158
317
792
Hazard warning
label
IRRITANT
IRRITANT
CORROSIVE
CORROSIVE
Procedure
Wear goggles (a face shield is preferable when handling large volumes) and chemical-resistant
gloves.
Work in a fume cupboard.
Measure out the indicated volume of concentrated nitric(V) acid in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in suitable beaker or laboratory jug.
Stir the solution (which will become warm).
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour the solution into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
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62
Although used in the past, tetrachloromethane (carbon tetrachloride) (see Hazcard 100) and
1,1,1-trichloroethane (see Hazcard 103), should no longer be chosen as the solvent. An alternative is
cyclohexane.
Hexanedioyl chloride (adipoyl chloride) may be used but is more unstable in storage, so it is normally better
to buy decanedioyl (sebacoyl) chloride.
The acid chlorides are liquids at room temperature. They can be weighed out in a suitable container.
Suppliers also provide the prepared solutions. These are used directly. Do check the labels.
General Hazards
The acid chlorides and hexane-1,6-diamine are CORROSIVE (see Hazcards 3B & 41)
and cyclohexane is HIGHLY FLAMMABLE & HARMFUL (see Hazcard 45B).
70
CLEAPSS 2011
63 Oscillating reactions
More information about these reactions can be found in Chemical Demonstrations (Vol 2), Shakashiri, 1985,
University of Wisconsin Press, ISBN 0299101304. The entire four volume series is a good source of
information.
CLEAPSS 2011
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64
Oxygen gas
Oxygen is prepared by adding hydrogen peroxide solution onto a catalyst, manganese(IV) oxide.
The reaction is very vigorous and exothermic and chemicals have been known to shoot up the thistle funnel
or the bung has been forced out of the flask. The rate of the reaction depends on the surface area of the
catalyst. It is better to use granules rather than a fine powder. If only powder is available, then only a small
amount is required.
Potassium iodide (1 g) produces oxygen at a steadier rate. The solution first saturates itself with oxygen and
then the gas is released.
Hydrogen peroxide decomposes slowly during storage. Suppliers add an inhibitor to slow the process, but
once this is used up, decomposition accelerates. If more hydrogen peroxide than expected is required, then
this is a sure sign that the hydrogen peroxide is old and new stock is required.
5 ml of 100 vol solution of hydrogen peroxide produces 500 ml of oxygen, ie, 5 x 100 ml.
General Hazards
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CLEAPSS 2011
65
Phosphoric(V) acid
Formula: H3PO4
General Hazards
Concentration
required
0.01 M
0.1 M
0.5 M
1M
1000
2500
Ten-fold dilution of the 0.01 M solution
3.5
7.0
17.0
17
34
85
34
68
170
Hazard warning
label
IRRITANT
Procedure
Wear goggles and chemical-resistant gloves.
Measure out the indicated volume of concentrated phosphoric(V) acid in a measuring cylinder.
Add the liquid to about two thirds of the final volume of water in a suitable beaker or laboratory jug.
Stir the solution.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour the solution into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
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66
Deficient in calcium:
Deficient in iron:
Deficient in magnesium:
Deficient in nitrogen:
Deficient in phosphorus:
Deficient in potassium:
Deficient in sulfur:
[CaH4(PO4)2.2H2O]
Solution 1
Dissolve the following salts in 1000 ml of pure water:
1 g of magnesium sulfate(VI)
1 g of potassium nitrate(V)
1 g of dipotassium hydrogenphosphate(V) [K2HPO4]
Solution 2
Dissolve 4 g of calcium nitrate(V) in 1000 ml of pure water.
Add solution 1 to solution 2 before use. This recipe is suitable for algae. For flowering plants, further dilution
by 2, 3 or 4 times may produce better results. This should be determined by experiment.
74
CLEAPSS 2011
67
These solutions are used to make buffer solutions (see Recipe Sheet 18 for Buffers). Masses are given to 3
decimal places but for many purposes, such accuracy is not required. Adjust to 2 decimal places.
Make sure you are using the correct salt. It is best to check with the formula (see below).
Sodium and potassium dihydrogen phosphate(V) solutions are best prepared just before use. During storage
there may be mould growth.
Potassium dihydrogen
phosphate(V)
Sodium dihydrogen
phosphate(V)
Sodium dihydrogen
phosphate(V)-1-water
KH2PO4
0.2 M
NaH2PO4
0.2 M
NaH2PO4.H2O
0.2 M
Dipotassium hydrogen
phosphate(V)
Dipotassium hydrogen
phosphate(V)-3-water
Disodium hydrogen
phosphate(V)
Disodium hydrogen
phosphate(V)-7-water
K2HPO4
0.2 M
K2HPO4.3H2O
0.2 M
Na2HPO4
0.2 M
Na2HPO4.7H20
0.2 M
Potassium
phosphate(V)
Sodium phosphate(V)
K3PO4
0.2 M
Na3PO4
0.2 M
Sodium phosphate(V)12-water
Na3PO4.12H2O
0.2 M
CLEAPSS 2011
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68
Potassium chloride
Molar mass: 74.55 g mol-1
Formula: KCl
100
400
Hazard warning
label
-
Procedure
Measure out the indicated quantity of potassium chloride.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
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CLEAPSS 2011
69
Potassium chromate(VI)
Molar mass: 194.19 g mol-1
Formula: K2CrO4
General Hazards
Hazard warning
label
TOXIC
TOXIC
VERY TOXIC
VERY TOXIC
Procedure
Wear chemical-resistant gloves and goggles. Use a fume cupboard if the solid is a fine powder.
Measure out the indicated quantity of potassium chromate(VI).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add the hazard warning.
CLEAPSS 2011
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70
Potassium dichromate(VI)
Potassium dichromate solutions are best made up in 0.1 M sulfuric(VI) acid rather than in pure water.
This can be used as acidified potassium dichromate(VI) solution.
The solution is used to test for the presence of alcohols. See Recipe Sheet 102 for Test reagents for organic
functional groups.
Molar mass: 294.18 g mol-1
Formula: K2Cr2O7
General Hazards
Hazard warning
label
TOXIC
TOXIC
VERY TOXIC
Procedure
Wear chemical-resistant gloves and goggles. Use a fume cupboard if the solid is a fine powder.
Measure out the indicated quantity of potassium dichromate(VI).
Add the solid to about two thirds of the final volume of 0.1 M sulfuric(VI) acid in a beaker or laboratory
jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
0.1 M sulfuric(VI) acid to the required level.
Pour into a labelled bottle. Add the hazard warning.
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CLEAPSS 2011
71
Potassium hydroxide
Solutions absorb carbon dioxide on standing. Solutions lower than 0.1 M are most significantly affected and
should, therefore, be freshly made.
Potassium hydroxide solution is best stored in plastic screw-cap bottles. It is known to etch glass when
stored for long periods and polystop caps can allow carbon dioxide to enter.
Equipment with glass joints, eg, burettes and bottles, can seize up over time.
Molar mass: 56.11 g mol-1
Formula: KOH
General Hazards
See Hazcard 91. When added to water so much heat is evolved that boiling could
occur.
Do not make this solution for the first time without seeking practical advice from a
more experienced colleague. A choking mist is often formed as the solid dissolves in
water. While this is not a serious safety risk, it is unpleasant and it is wise to use a
fume cupboard where possible. If saturated potassium hydroxide has to be prepared
after carrying out an exhaustive risk assessment, then start from 5 M potassium
hydroxide solution and add pellets a little at a time. Do not store this solution.
Hazard warning
label
IRRITANT
IRRITANT
CORROSIVE
CORROSIVE
Procedure
Wear chemical-resistant gloves and goggles. Use a fume cupboard if the solid is a fine powder.
Measure out the indicated quantity of potassium hydroxide.
Add the solid in stages to about two thirds of the final volume of water in a beaker or laboratory jug. If
concentrated solutions are being made, ice should be used in place of water.
Stir carefully to dissolve before adding the next group of pellets. It may be necessary to cool the
solution between additions. (Ice could be added.)
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled plastic bottle. Add a hazard warning if appropriate.
Wear chemical-resistant gloves and goggles. Use a fume cupboard if the solid is a fine powder.
Measure out 10 g of potassium hydroxide.
Place 70 ml of the alcohol (ethanol or methanol) in a glass 250 ml beaker.
Insert a magnetic stirrer bar and place the beaker on a magnetic stirrer.
Add the potassium hydroxide solution in 3 parts, allowing the solid to dissolving before adding the next
amount. The process is exothermic but not as extreme as that with water.
Place the solution in a 100 ml measuring cylinder and make up to the 100 ml mark with the alcohol.
Pour into a labelled plastic bottle. Label the solution CORROSIVE and HIGHLY FLAMMABLE.
CLEAPSS 2011
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72
Potassium iodide
Solutions of potassium iodide often go yellow on keeping. If this occurs, add 0.1 M sodium thiosulfate
solution dropwise, or a crystal of the solid with stirring, until the solution becomes colourless.
Formula: KI
Hazard warning
label
-
Procedure
Wear eye protection.
Measure out the indicated quantity of potassium iodide.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
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CLEAPSS 2011
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Potassium manganate(VII)
Formula: KmnO4
General Hazards
0.1 M
0.02 M
0.1 M
Saturated
0.1 M
1M
1M
Hazard
warning label
IRRITANT
IRRITANT
Procedure
Wear eye protection.
It would be advisable to wear gloves as the chemical stains the skin brown.
Measure out the indicated quantity of potassium manganate(VII).
Add the solid to about two thirds of the final volume of the suggested dilute sulfuric(VI) acid in a
beaker or laboratory jug.
Stir to dissolve. It is better not to heat but dissolving can take some time, and it is difficult to see when
the solid has all dissolved.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
dilute sulfuric(VI) acid to the required level.
Pour into a labelled bottle.
CLEAPSS 2011
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74
Preservatives prevent chemical decomposition of the fixed material (see Recipe Sheet 41).
More information about the procedures for fixing and preserving specimens can be found in the CLEAPSS
Handbook, section 15.8.
Preserving animal
tissue (Recipe 1)
Preserving animal
tissue (Recipe 2)
Kaiserlings
preservative
82
CLEAPSS 2011
75
Saline solutions are suitable for temporary microscopial preparations and short experiments.
For more prolonged investigations, a suitable Ringers solution should be used.
Ringers solutions
For frogs
Dissolve 6.50 g of sodium chloride, 0.12 g of calcium chloride-6-water (IRRITANT), 0.14 g of
potassium chloride and 0.20 g of sodium hydrogencarbonate in 1000 ml of water.
For mammals
Dissolve 8 g of sodium chloride, 0.2 g of calcium chloride-6-water (IRRITANT), 0.2 g of
potassium chloride, 0.19 g of magnesium chloride-6-water, 0.05 g of sodium
dihydrogenphosphate and 1 g of sodium hydrogencarbonate in 1000 ml of water.
For locusts
Dissolve 7.60 g of sodium chloride, 0.22 g of calcium chloride-6-water, 0.75 g of potassium
and other
chloride, 0.19 g of magnesium chloride-6-water, 0.48 g of sodium dihydrogen phosphate,
insects
and 0.37 g of sodium hydrogencarbonate in 1000 ml of water.
For
Dissolve 6 g of sodium chloride, 0.2 g of calcium chloride-6-water, 0.12 g of potassium
earthworms
chloride and 0.1 g of sodium hydrogencarbonate in 1000 ml of water.
For marine
Dissolve 31 g of sodium chloride, 1.37 g of calcium chloride-6-water (IRRITANT), 0.99 g of
crustaceans
potassium chloride, 2.35 g of magnesium chloride-6-water, 0.48 g of sodium
dihydrogenphosphate, and 0.22 g of sodium hydrogencarbonate in 1000 ml of water.
Saline solutions
For invertebrate tissues
For amphibian tissues
For mammalian tissues
For mammalian blood
CLEAPSS 2011
83
76
Sandells solution
84
CLEAPSS 2011
77
Silver nitrate(V)
This is probably the most expensive chemical used in schools; use it with care, retrieving unused stock.
Pure water (ie, distilled or deionised) should be used to avoid cloudiness of solutions.
Tollens reagent [ammoniacal silver nitrate(V) solution], (see Recipe sheet 102) should not be stored. It
should be made and disposed of in situ by the students.
Molar mass: 169.87 g mol-1
Formula: AgNO3
General Hazards
See Hazcard 87. The solid and more-concentrated solutions will stain skin and other
organic material black. This stain is difficult to remove. Work surfaces also stain.
Hazard warning
label
IRRITANT
CORROSIVE
CORROSIVE
Procedure
Wear eye protection.
Measure out the indicated quantity of silver nitrate(V).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add a hazard warning if appropriate.
CLEAPSS 2011
85
78
Slime
Different sources of polyvinyl alcohol vary in their average molar mass; some are about 115000 g mol-1,
while others are much less, at around 17000 g mol-1 (and less expensive). The larger the average molar
mass, the better the slime.
PVA wood glue is cheaper still but contains polyvinyl acetate and other chemicals such as binders. This will
affect the nature of the slime and it may become more solid.
Custard powder can also be used with PVA glue to form bouncing custard balls.
Slime is a gel. It is thought that borate ions react with hydroxyl groups in the polymer of vinyl alcohol to form
cross links, with the elimination of water. These cross links probably involve hydrogen bonds which
continually form and break under flow.
The sliminess of the gel can be adjusted by altering the amount of sodium borate used. Viscosity changes
can be detected by timing the passage of a ball bearing through the slime in a measuring cylinder.
If acid is added to the slime, the gel collapses to give a free-flowing solution.
Although the slime can be stored in a labelled sealed bag and placed in a refrigerator, it can develop mould if
kept for long periods.
Disposal: Add enough 1 M sulfuric(VI) acid to just allow the solution to become free-flowing, dilute and pour
down a foul-water drain.
These chemicals, and the made-up slime, should not be taken home.
General Hazards
See Hazcard 14. However, since publication of Hazcards, the toxicity of boron
compounds has been the subject of a considerable review and debate in the
European Union. Some suppliers will now label the bottle TOXIC with R63 (Possible
risk of harm to the unborn child) warning. However only solutions above 8% (w/v)
will be labelled TOXIC.
Sodium tetraborate should be weighed in a fume cupboard to avoid raising dust.
86
CLEAPSS 2011
79
Alcoholic soap solution can be titrated against tap water in a stoppered bottle until a froth, stable for 15
seconds, is obtained. The concentration of soap solution and/or volume of water involved in the titration will
depend on the hardness of the local tap water. Solutions should be tested before being presented to a class
so that minor adjustments to the formulation can be made.
The alcoholic soap solution is called either Clarkes or Wanklyns soap solution.
General Hazards
See Hazcards 37 & 40A. Do not heat soap solutions made up in ethanol with a
naked flame. Use a hot plate or surround the beaker with hot water.
Bubble mixture
Recipes may differ with different makes of liquid detergent.
The recipe below works best using Fairy Liquid. (other detergents will also work but you may need to
experiment with quantities.
Procedure to make a bubble solution
Mix together by volume, 10 parts liquid detergent, 85 parts pure water and 5 parts propane-1,2,3-triol
(glycerol).
CLEAPSS 2011
87
80
Sodium carbonate
Formula: Na2CO3
Formula: Na2CO3.10H2O
General Hazards
Hazard warning
label
IRRITANT
Hazard warning
label
IRRITANT
Procedure
Wear eye protection.
Measure out the indicated quantity of anhydrous or hydrated sodium carbonate.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add a hazard warning if appropriate.
88
CLEAPSS 2011
81
Formula: NaClO
General Hazards
Concentration required
1%
10000 ppm
0.15 M
0.25%
2500 ppm
0.038 M
0.1%
1000 ppm
0.015 M
0.01%
100 ppm
0.0015 M
Preparation
See below for procedure.
No need for further dilution.
Mix, by volume, 1 part of the 1%
solution with 3 parts of water.
Mix by volume, 1 part of the 1%
solution with 9 parts of water.
Dilute the 0.1% solution further
by mixing by volume, 1 part of
the 0.1% solution with 9 parts of
water.
Use
Disinfectant for blood spills or
dirty conditions.
Disinfectant for micro-biological
discard pots.
Disinfectant for general use, eg,
sterilising solution.
Disinfectant for sterilising
mouthpieces, swabbing skin.
CLEAPSS 2011
89
82
Sodium chloride
Molar mass: 58.44 g mol-1
Formula: NaCl
Mass (g) of solid to be used
Concentration
required
100
0.1 M
0.5 M
1.0 M
Saturated (20 C)
2.92
5.84
37
Hazard warning
label
-
Procedure
Measure out the indicated quantity of sodium chloride.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
90
CLEAPSS 2011
83
Sodium ethanoate
Formula: NaCH3COO
Mass (g) of solid to be used
Concentration
required
100
0.2 M
1.0 M
Saturated (20 C)
8.20
50
Hazard warning
label
-
Formula: NaCH3COO.3H2O
Mass (g) of solid to be used
Concentration
required
100
0.2 M
1.0 M
Saturated (20 C)
Hazard warning
label
-
Procedure
Measure out the indicated quantity of sodium ethanoate.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
CLEAPSS 2011
91
84
Sodium hydrogencarbonate
Molar mass: 84.01 g mol-1
Formula: NaHCO3
Mass (g) of solid to be used
Concentration
required
100
0.1 M
0.5
1.0 M
Saturated (20 C)
Hazard warning
label
-
Procedure
Measure out the indicated quantity of sodium hydrogencarbonate.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve. Do not warm as decomposition may take place.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle.
Procedure
Wear eye protection when preparing the solution.
Prepare a saturated solution of sodium hydrogencarbonate using freshly-boiled pure water.
Place a wide-bore glass tube into the water and then pour liquid paraffin onto the waters surface.
Add a few crystals of benzene-1,2,3-triol (pyrogallol) down the glass tube and then slowly withdraw it.
(1 g of benzene-1,2,3-triol is capable of absorbing 190 cm3 of oxygen.)
92
CLEAPSS 2011
85
Sodium hydroxide
Solutions absorb carbon dioxide on standing. Solutions lower than 0.1 M are most significantly affected and
should, therefore, be freshly made.
Sodium hydroxide solution is better stored in plastic screw-cap bottles. It is know to etch glass when stored
for long periods and polystop caps can allow carbon dioxide to enter.
Equipment with glass joints, eg, burettes and bottles, can seize up over time.
Formula: NaOH
General Hazards
See Hazcard 91. When added to water, heat is evolved such that boiling could
occur. Do not make this solution for the first time without seeking practical advice
from a more-experienced colleague. A choking mist is often formed as the solid
dissolves in water. While this is not a serious safety risk, it is unpleasant and it is
wise to use a fume cupboard where possible. If after carrying out an exhaustive risk
assessment, there is no alternative to preparing saturated sodium hydroxide, then
start from 5 M sodium hydroxide solution and add pellets a little at a time. Do not
store this solution.
Hazard warning
label
IRRITANT
IRRITANT
CORROSIVE
CORROSIVE
Procedure
Wear chemical-resistant gloves and goggles. Use a fume cupboard if the solid is a fine powder.
Measure out the indicated quantity of sodium hydroxide.
Add the solid in stages to about two thirds of the final volume of water in a beaker or laboratory jug. If
concentrated solutions are being made, ice should be used in place of water.
Stir carefully to dissolve before adding the next group of pellets. It may be necessary to cool the
solution between additions. (Ice could be added.)
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour into a labelled bottle. Add a hazard warning if appropriate.
CLEAPSS 2011
93
86
See Hazcard 95B. The solution is strongly alkaline. Consult the relevant Hazcards
when using metal salts to add to the chemical garden.
Procedure to make 250 ml of sodium silicate solution for a chemical garden from the
commercially-available solution
Wear eye protection.
Add 200 ml of commercial sodium silicate to 600 ml of pure water in a 1-litre beaker.
Pour into a labelled bottle. Although the solution is deemed low hazard, it might be prudent to label the
bottle IRRITANT as the solution is strongly alkaline.
Procedure to make 250 ml of 8% sodium silicate solution from the solid
Wear eye protection.
Add 20 g of sodium silicate to a 600 ml beaker and add 250 ml of pure water.
Add a magnetic stirrer bar and place the mixture on a heater/stirrer.
Maintain a temperature of about 45 C until the solid appears to dissolve.
Filter through Whatman No.1 filter paper.
Pour into a labelled bottle. Although the solution is deemed low hazard, it might be prudent to label the
bottle IRRITANT as the solution is strongly alkaline.
The crystal (chemical) garden
Wear eye protection and consider wearing disposable nitrile gloves.
Pour the prepared sodium silicate into a 600 ml beaker (a labelled coffee jar is also suitable), cover it
with a clock glass or sheet of glass (check edges are not sharp) and allow the liquid to settle. (After
adding crystals, it is better not to move the container so any moving should be done at this stage.)
Now add small (eg, rice-grain size) crystals of manganese(II) sulfate(VI), copper(II) sulfate(VI),
chromium(III) chloride, iron(II) sulfate(VI), iron(III) chloride, cobalt(II) chloride, tin(II) chloride,
aluminium sulfate(VI), nickel(II) sulfate(VI) or any other soluble salt you wish to try.
Leave the container undisturbed for several days. Label the container IRRITANT.
Growing lead(II) iodide crystals in gels
Dilute 16 ml of commercial sodium silicate solution to 100 ml with pure water (solution A).
Place 15 ml of 1 M ethanoic acid (see Recipe Sheet 39) in a beaker and add 0.67 g of potassium
iodide (solution B).
Add 15 ml of solution A to solution B, stirring vigorously. Pour the mixture into a boiling tube and allow
it to set (5 to 60 minutes).
Dissolve 0.66 g of lead(II) nitrate(V) in 2 ml of water and pour this onto the top of the gel. Label the
container IRRITANT.
Observe the spread of crystals of lead(II) iodide over the next few weeks.
94
CLEAPSS 2011
87
Sodium thiosulfate
Formula: Na2S2O3.5H20
Mass (g) of solid to be used
Concentration
required
100
0.01 M
0.1 M
0.5 M
1.0 M
Saturated (20 C)
Hazard warning
label
-
Procedure
Measure out the indicated quantity of sodium thiosulfate-5-water.
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve. Do not warm the solution.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Add about 1 g of sodium hydrogencarbonate or sodium sulfate(IV) (sulfite) (HARMFUL) for every
1000 ml of solution, to help to preserve it.
Pour the solution into a labelled bottle and mix well.
A saturated solution
The saturated solution requires little water. Weigh out the sodium thiosulfate as indicated and add it to hot
(almost boiling) water with a volume in millilitres equal to about half the weight of solid in grams. Stir to
dissolve and leave to cool to room temperature. If there is no solid present in the cooled solution, add one
crystal of solid to it. If this causes further solid to form, then the solution is saturated and may be left in
contact with the solid. If not, add further solid.
CLEAPSS 2011
95
88
Gram Stain
For details on
using this stain
see Guidance
Leaflet GL 95
96
CLEAPSS 2011
89
Aceto carmine
Ehrlichs
haematoxylin
for animal
histology
Eosin for
cytoplasmic
staining
Feulgen's stain
for DNA
Leishman's
stain for blood
cells
Methyl green
pyronin for DNA
and RNA
Methylene blue
for cheek cells
Orcein acetic
(Acetic orcein)
for
chromosomes
Sudan III or IV
solutions for
lipids
CLEAPSS 2011
97
90
Methylene blue is less sensitive than Azure A and not so convenient to use in electrophoresis but it is
cheaper and adequate for some activities.
Azure A for DNA
Colloidal
Coomassie Blue
for proteins
Methylene blue
for DNA
Low hazard.
Low hazard.
www.ncbe.reading.ac.uk/NCBE/SCIENCEYEAR/recipes.html
98
CLEAPSS 2011
91
Lactophenol
Cotton blue
(aniline blue) for
fungi
CLEAPSS 2011
99
92
Janus green B
(Diazine green)
for mitochondria
Methylene blue
for cell contents
Neutral red
Resazurin
solution to test
for milk
freshness
TTC, to show
respiratory
activity
100
Resazurin is IRRITANT.
Low hazard.
CLEAPSS 2011
Iodine stain
Phloroglucinol
for pentoses
and lignin
Schulzes
solution for
cellulose
CLEAPSS 2011
101
For phenylammonium
sulfate(VI): see Hazcard 4.
Wear eye protection and
disposable nitrile gloves when
making up the solution.
Label the stain HIGHLY
FLAMMABLE.
For lactophenol, see
Hazcard 38C; for iodine, see
Hazcard 54.
Wear goggles and chemicalresistant gloves.
Label the stain TOXIC.
See Recipe Sheet 50.
Phloroglucinol is an IRRITANT.
Ethanol is HIGHLY FLAMMABLE
(see Hazcards 12, 40 and 47).
Label solution HIGHLY
FLAMMABLE. Wear eye
protection.
Zinc chloride is CORROSIVE and
iodine is HARMFUL (see
Hazcards 54 and 108). Wear
eye protection, and chemicalresistant gloves, and carry out
the procedure in a fume
cupboard. Label the solution
CORROSIVE.
94
A primary standard solution is one with a concentration that is accurately known. (They do not have to be
0.1 M) They are used to determine the concentration of other reagents undergoing titration.
Solids used should be of high purity (ie, more than 99.5% pure). Hydrated salts should not be used.
The solid should not alter composition during weighing (which is why sodium hydroxide is not used; it
absorbs water and carbon dioxide).
Distilled or deionised water must be used.
If possible, use a balance reading to 3 decimal places.
102
CLEAPSS 2011
CLEAPSS 2011
103
95
Strontium chloride
NB:
If a school has only strontium carbonate, the solution can be made by adding 1.48 g of the carbonate to
100 ml of 0.1 M hydrochloric acid.
104
CLEAPSS 2011
96 Sulfur dioxide
For acid rain see the Recipe sheet 97.
This is a particularly nasty gas to prepare and use. Do make sure that the procedure is trialled with help from
a more-experienced colleague and that the fume cupboard is working satisfactorily.
General Hazards
See Hazcards 47A, 78, 92 & 97. Sulfur dioxide is TOXIC by inhalation. There is no
safe Workplace Exposure Limit for sulfur dioxide. As a guide, the HSE has advised
that levels, must be less than 2.7 mg m-3 (ie, 1 ppm). Any preparation of sulfur
dioxide should be carried out in a fume cupboard.
Acidified potassium
dichromate(VI) paper
Loose mineral
wool plug
Heat
CLEAPSS 2011
105
97
Sulfur dioxide is a particularly nasty gas to prepare and use. Do make sure that the procedure is trialled with
help from a more-experienced colleague and that the fume cupboard is working satisfactorily.
General Hazards
See Hazcards 47A, 78, 92 & 97. Sulfur dioxide is TOXIC by inhalation. There is no
safe Workplace Exposure Limit for sulfur dioxide. As a guide, the HSE has advised
that levels must be less than 2.7 mg m-3 (ie, 1 ppm). Any preparation of sulfur
dioxide should be carried out in the fume cupboard.
Method 1
Use a fume cupboard. Make sure it is working Wear goggles.
3
Place between 10 and 20 g of sodium metabisulfite (HARMFUL) into a 250 cm conical flask.
Set up the equipment as shown below. The inverted filter funnel should be held just below the surface
of the water in the beaker.
Pour enough 2 M hydrochloric acid (IRRITANT) to just cover the bottom of the thistle funnel tube in the
flask.
Heat the conical flask gently to produce the gas, which will dissolve as it meets the water in the
beaker. The inverted funnel prevents suck back.
Add 5 M hydrochloric acid if more sulfur dioxide is required.
The leftover reagents can be poured down the fume cupboard sink with plenty of water.
Add at least 50 cm3 of
2 M hydrochloric acid
Inverted funnel
just below water
surface
Heat
Method 2
In a fume cupboard, dissolve 9.5 g of sodium metabisulfite (HARMFUL) in 100 ml of water. Add 100 ml of
0.5 M sulfuric(VI) acid and make up to 250 ml.
Acid Rain
Pupils should open containers in a fume cupboard and particular attention should be paid to known
asthmatics.
For acid rain, use 0.0001 M sulfuric(VI) acid rather than a sulphur dioxide solution.
For an acidic atmosphere, use a 0.1 M solution of sodium metabisulfite in water. This will generate enough
SO2 gas to provide an acid atmosphere.
106
CLEAPSS 2011
98
Sulfuric(VI) acid
The procedure below uses ice made from distilled or deionised water. Make sure you have a supply of this in
the freezer.
Do not make dilute solutions for the first time without seeking practical advice from a more-experienced
colleague.
Formula: H2SO4
General Hazards
Concentration
required
0.01 M
0.1 M
0.4 M
1M
Battery acid
5M
Hazard warning
label
IRRITANT
CORROSIVE
CORROSIVE
Procedure
Wear goggles (a face shield is preferable when handling large volumes) and chemical-resistant
gloves.
Measure out the indicated volume of concentrated sulfuric(VI) acid in a measuring cylinder.
Fill the beaker or laboratory jug half to two thirds full with ice, add 200 ml of water and a stirrer bar.
Set the stirrer running on a magnetic stirrer and add the concentrated sulfuric(VI) acid slowly onto the
ice.
Keep stirring the solution until the ice melts.
Pour the solution from the beaker into an appropriate measuring cylinder or laboratory jug and add
water to the required level.
Pour the solution into a labelled bottle and mix well. Add a hazard warning if appropriate.
CLEAPSS 2011
107
99
Ammonia
Use damp red litmus
paper
Place concentrated
hydrochloric acid close by
Carbon dioxide
Use lime water
Carbon monoxide
Light it in a fume
cupboard
Chlorine
Use damp blue litmus
paper
Dinitrogen monoxide
Add a lighted or glowing
splint
Hydrogen
Add a lighted splint
Hydrogen chloride
Use damp blue litmus
paper
Place concentrated
ammonia solution close
by
Hydrogen sulfide
Moisten filter paper with
0.05 M lead(II) nitrate(V)
solution
Nitrogen
There is no positive test for nitrogen.
Nitrogen dioxide
Observe the colour
Nitrogen monoxide
Add the gas to air
Oxygen
Add a glowing splint
Sulfur dioxide
Moisten filter paper with
0.05 M acidified
potassium dichromate(VI)
solution
108
CLEAPSS 2011
Carbonate ions
2 M hydrochloric acid
Chloride ions
0.05 M silver nitrate(V)
solution
Iodide ions
0.05 M silver nitrate(V)
solution
Wear goggles.
See Hazcards 55B &
98.
See Hazcards 6, 8, 9A
& 67.
Nitrate(V) ions
Brown ring test
Devadas alloy
Wear goggles.
See Hazcard 1 & 91.
Phosphate ions
0.5 M ammonium
molybdate(VI) solution
(HARMFUL)
Sulfate(VI) ions
0.1 M barium chloride
solution (HARMFUL)
CLEAPSS 2011
109
2 Add 0.4 M
sodium
hydroxide to a
small amount of
the solution
under test
3 0.1 M sodium,
potassium or
ammonium
sulphide
solution
Aluminium ions
Alizarin solution
Ammonium ions
0.1 M sodium
hexanitrocobalt-ate(III)
solution (HARMFUL)
0.4 M sodium
hydroxide solution
(IRRITANT)
Calcium ions
0.25 M ammonium
ethanedioate
110
CLEAPSS 2011
Cobalt(II) ions
Thiooxamide
(rubeanic acid)
(HIGHLY FLAMMABLE,
HARMFUL)
0.1 M ammonium or
potassium thiocyanate
See Hazcard 9B or
95C.
See Hazcards 6.
Prepare in a fume
cupboard.
See Hazcards 35 and
also 40A.
See Hazcard 9B or
95C. The solution is
LOW HAZARD.
0.1 M potassium
hexacyanoferrate(III)
1,10 phenanthroline
solution
Copper(II) ions
2M ammonia solution
Thiooxamide
(rubeanic acid)
(HIGHLY FLAMMABLE,
HARMFUL)
0.1 M ammonium or
potassium thiocyanate
Iron(II) ions
Iron(III) ions
0.1 M potassium
hexacyanoferrate(III)
Ferroxyl indicator
(HIGHLY FLAMMABLE)
0.1 M ammonium or
potassium thiocyanate
Lead(II) ions
Dithizone
Potassium iodide
CLEAPSS 2011
111
Nickel(II) ions
Thiooxamide
(rubeanic acid)
(HIGHLY FLAMMABLE,
HARMFUL)
Butanedione dioxime
(dimethylglyoxime)
(HIGHLY FLAMMABLE &
HARMFUL)
See Hazcards 6.
Prepare in a fume
cupboard. The solution
is LOW HAZARD.
Potassium ions
0.1 M Sodium
hexanitrocobalt-ate(III)
(HARMFUL)
Zinc(II) ions
2M ammonia solution
112
CLEAPSS 2011
Potassium
dichromate(VI) test
Potassium
manganate(VII)
test
See Hazcards
47A & 108A.
Schiffs reagent
Carboxylic acids
Neutral iron(III)
chloride
CLEAPSS 2011
113
See Hazcards 81
and 98.
Phenol
Neutral iron(III)
chloride
114
CLEAPSS 2011
Formula: SnCl2.2H20
General Hazards
Hazard warning
label
-
Procedure
Wear goggles and chemical-resistant gloves. Use a fume cupboard.
Measure out the indicated quantity of tin(II) chloride-2-water.
Add the solid to about 10% of the final volume of concentrated hydrochloric acid in a beaker.
Stir to dissolve. Do not warm the solution.
Pour the solution into a measuring cylinder or laboratory jug and dilute to the final volume with
distilled/deionised water.
Leave to clear.
Pour the solution into a labelled bottle and mix well.
CLEAPSS 2011
115
A.
Sea water
Dissolve the following salts in 250 ml of pure water and dilute to 1000 ml.
B.
C.
Measure 0.45 g of calcium sulfate(VI)-2-water and add it to 1000 ml of water in a bottle and leave it
overnight to dissolve.
Filter 130 ml of freshly-made calcium hydroxide solution (see Recipe Sheet 20).
Pass carbon dioxide though the solution so that calcium carbonate first precipitates and then, with
further carbon dioxide, re-dissolves to form calcium hydrogencarbonate. This may take some time.
Dilute this clear solution to 1000 ml.
116
CLEAPSS 2011
See Hazcards 60, 91 & 98. Great care must be taken in making the 8 M sodium
hydroxide solution. Gloves and eye protection (a face shield is preferable) should be
used. The preparation is very slow and must not be rushed.
The indicator
Prepare 0.01 M sodium thiosulfate solution (see Recipe Sheet 87) which can be
standardised against potassium iodate solution (see Recipe Sheet 94) for
accurate work.
Prepare 1% starch solution. See Recipe Sheet 49.
CLEAPSS 2011
117
Formula: ZnSO4.7H2O
General Hazards
Hazard warning
label
HARMFUL
HARMFUL
Procedure
Measure out the indicated quantity of hydrated zinc sulfate(VI).
Add the solid to about two thirds of the final volume of water in a beaker or laboratory jug.
Stir to dissolve, warming if necessary.
Either pour the solution from the beaker into an appropriate measuring cylinder and add water to the
required level or add water to the required level in the laboratory jug.
Pour into a labelled bottle.
118
CLEAPSS 2011
Index
Name
Name
Recipe
Sheet
Recipe
Sheet
Acetic acid
39
Aniline blue
91
Acetic alcohol
41
Aniline sulfate(VI)
93
Acetic orcein
89
74
Aceto carmine
89
Acid atmosphere
97
Animal tissue
preservative
Anion tests
100
46
Aspirator
20
Acid rain
97
Azo dyes
Agar
Azure A
90
Alcohol tests
102
Alcohol/water solutions
Alcoholic potassium
hydroxide
Alcoholic solutions
71
2
Aldehyde tests
102
Alginate beads
Alizarin solution
101
Alkaline pyrogallol
84
Alum
Aluminium chloride
Aluminium etching
38
Aluminium potassium
sulfate(VI)
Aluminium solutions
Amino acid
chromatography
Amino acid tests
26
Barium chloride
10
49
Barium nitrate(V)
10
Barium solutions
10
Basic agar
Belousov-Zhabotinski
reaction
Benedicts qualitative
reagent
63
Benedicts quantitative
reagent
Bile salts
12
Biological buffers
18
Biro ink
chromatography
Bismuth nitrate(V)
26
11
37
14
Biuret reagent
15
13
Bleach
81
Ammonia gas
Blue-bottle reaction
63
Ammonia solution
Blueprints
52
Ammonia test
99
B-R reaction
63
Ammonium cerium(IV)
sulfate(VI)
22
Bradys reagent
33
Briggs-Rausher reaction
63
Ammonium chloride
Brodies fluid
16
100
Ammonium
ethanedioate
Ammonium hydroxide
101
Ammonium iron(III)
citrate
Ammonium iron(III)
sulfate(VI)
Ammonium sulfide
52
101
Ammonium thiocyanate
101
52
Ammonium vanadate
Amylase
37
Analgesics
chromatography
Andrews arrangement
for gas preparation
26
CLEAPSS 2011
24
119
Bromine water
17
Bromocresol green
46
Bromophenol blue
46
Bubble mixture
79
Buffers
18
Butanedione dioxime
101
B-Z reaction
63
Calcium carbonate
(standard solution)
Calcium chloride
94
19
Calcium hydroxide
20
Calcium nitrate(V)
19
Name
Recipe
Sheet
Recipe
Sheet
Carbohydrate tests
13
Cotton blue
91
Carbon dioxide
99
Cotton dying
21
Cresol red
46
48
32
Carbon monoxide
preparation
Carbon monoxide test
42
Cytological fixative
41
99
DCPIP
13
100
51
33
Diammonium iron(II)
sulfate(VI)
Diazine green
102
Diazonium salts
Cation tests
101
13
Cellular respiration
13
dichlorophenol indophenol
Cerium(IV) solutions
22
49
Cerium(IV) sulfate(VI)
22
Dichromate(VI) titration
indicators
Cheek cells
89
Dimethylglyoxime
101
Chemical garden
86
42
Chemiluminesence
23
100
Dinitrogen monoxide
preparation
Dinitrogen monoxide
test
Dinitrophenylhydrazine
33
Chlorine gas
preparation
Chlorine test
24
Dinitrosalicylic acid
34
Dipotassium hydrogen
phosphate(V)
67
Chlorine water
25
67
Chlorophyll
chromatography
Chromatography
solvents
Chrome alum
26
Dipotassium hydrogen
phosphate(V) -3- water
Disappearing cross
reaction
Disinfectants
29
27
67
Chromium(III) chloride
27
Disodium hydrogen
phosphate(V)
Citric acid
28
Disodium hydrogen
phosphate(V) -7- water
67
2,6-
2,3,53,5-
99
26
92
99
81
Clarkes fluid
41
Dissolved oxygen
105
79
Dithizone
101
Clock reactions
29
DNP
33
Cobalt(II) chloride
30
Cobalt(II) nitrate(V)
30
DNS
34
Cobalt(II) thiocyanate
30
DNSA
34
Coles reagent
13
102
Colloidal Coomassie
blue
Complexometric
indicators
Coomassie blue
90
35
Dyes
Dying cotton
49
90
Ehrlichs haematoxylin
89
Copper electroplating
36
Eiosin indicator
49
Copper etching
38
Electrolytic agar
Copper(II) chloride
31
Enzymes
37
Copper(II) nitrate(V)
31
Enzymes immobilised
Copper(II) solutions
31
Eosin
89
Copper(II) sulfate(VI)
31
Eriochrome black
49
2,4-
49
120
CLEAPSS 2011
Recipe
Sheet
Etching solutions
38
Indicator agar
100
100
Ethanoic acid
39
Iodine solution
50
Ethanol/water mixtures
Iodine stain
93
Ethanolic potassium
hydroxide
71
Iodoform test
102
Iron etching
38
Iron(II) chloride
51
Iron(II) solutions
51
Iron(II) sulfate(VI)
51
FAA
2-
Name
Recipe
Sheet
41, 74
FABIL
93
Fehling's solution
40
Ferroin indicator
49
Ferroxyl agar
Iron(III) chloride
52
29
Ferroxyl indicator
101
Feulgen's stain
89
Iron(III) solutions
52
Fixatives in biology
41
Iron(III) sulfate(VI)
52
Flame tests
101
Janus green B
Fluorescein indicator
49
Formaldehyde
58
41
Kaiserlings
preservative
Ketone tests
Formalin-aceto fixative
41
Formalin-aceto-alcohol
74
Formic acid
59
Glucose quantitative
test
Glucose test
12
11, 34
13, 92
74
102
66
Lactophenol
91
29
Lead etching
38
Lead nitrate(V)
53
Leishman's stain
89
Lime water
20
Gram Stain
88
Lipase
37
Harcourt-Essen reaction
29
26
Hard water
104
Lipstick
chromatography
Lithium chloride
Home-made indicators
46
Litmus
46
Hydrochloric acid
43
Locating agents
26
Hydrogen
99
Lucas reagent
102
Hydrogen carbonate
indicator
Hydrogen chloride
preparation
Hydrogen chloride test
48
Lugols iodine
88
Luminol
23
Magnesium sulfate(VI)
55
99
Malt agar
Hydrogen gas
44
56
Hydrogen peroxide
45
Manganese(II)
sulfate(VI)
Manometric fluid
Hydrogen peroxide/
potassium iodide
reaction
Hydrogen sulfide
preparation
Hydrogen sulfide test
29
Mayonnaise agar
Mercury(I) nitrate(V)
57
42
Mercury(II) chloride
57
Hydrogen-sulfite test
102
Metal ion
chromatography
Methanal solution
26
99
58
hydroxy-1,2,3tricarboxylic acid
27
100
Methanoic acid
59
Immobilised enzymes
Methanol solution
CLEAPSS 2011
42
121
54
16
71
Methyl benzoate
nitration
Methyl green pyronin
26
89
Methyl orange
46
Methyl orange
preparation
Methyl red
Methylene blue
Milk agar
46
13, 89, 90,
92
1
Millons reagent
13
Mohrs salt
51
Molischs solution
13
Murexide
49
Neutral Red
46, 92
102
Nickel electroplating
36
Nickel(II) sulfate(VI)
60
Ninhydrin
13
100
Nitrate(V) test
100
Nitration of Methyl
benzoate
Nitric(V) acid
26
Nitrogen preparation
42
Nitrogen test
99
Nitrogen dioxide
preparation
Nitrogen dioxide test
42
61
99
63
Nitrogen monoxide
preparation
Nitrogen monoxide test
42
99
N-phenylanthranilic acid
indicator
49
1
62
OH group test
102
Orange II preparation
Name
Recipe
Sheet
Orcein acetic
90
Organic functional
group tests
Orthophosphoric acid
102
Oscillating reactions
63
Oxygen preparation
64
Oxygen test
99
65
105
PAS reaction
13
Pepsin
37
13
104
18
pH probe calibration
buffers
Phenanthroline solution
101
Phenol red
46
phenolindodichlorophenol
Phenolphthalein
13
18
46
Phenolphthalein
indicator agar
Phenylammonium
sulfate(VI)
Phenylthiocarbamide
13
Phenylthiourea
13
Phloroglucinol
93
100
Phosphoric(V) acid
65
Phosphorus
pentachloride
Physiological saline
solutions
Plant mineral
requirement solutions
102
Plant tissue
preservative
Polypeptide tests
74
Potassium chloride
68
Potassium chromate
69
Potassium
dichromate(VI)
Potassium dihydrogen
phosphate(V)
70
Potassium
hexacyanoferrate(III)
Potassium hydrogen
phthalate (standard
solution)
Potassium hydroxide
101
Potassium iodate
(standard solution)
Potassium
iodate/sodium
metabisulfite reaction
Potassium iodide
94
Potassium
iodide/potassium
persulfate clock reaction
122
Recipe
Sheet
1
93
75
66
13
67
94
71
29
72, 101
29
CLEAPSS 2011
94
Sodium
hexanitrocobaltate(III)
Sodium
hydrogencarbonate
Sodium hydroxide
101
101
Sodium hypochlorate
81
13
Sodium phosphate(V)
67
13
Sodium phosphate(V)
-12- water
67
101
Potassium thiocyanate
101
Precipitation indicators
49
74
Propanone solution
Protein tests
73
67
PTU
13
PVA
78
Pyrogallol
84
46
Redox indicators
Reducing sugar test
Recipe
Sheet
Sodium ethandioate
(standard solution)
Sodium ethanoate
Potassium
manganate(VII)
Potassium
phosphate(V)
Potassium sulfide
PTC
Name
Recipe
Sheet
83
84
85
Sodium silicate
86
Sodium sulfide
101
37
49
Sodium
tauroglycocholate
Sodium thiosulfate
11, 12
87
Solochrome black
49
Resazurin solution
93
89
Ringers solution
75
101
66
Safranin counterstain
88
Sakaguchi test
13
Saliva
37
Stains for
electrophoresis
Stains for fungal
material
Stains for metabolic
activity
Stains for plant material
90
Rubeanic acid
93
Sandells solution
76
Standard solutions
94
Schiffs reagent
102
Starch agar
Schulzes solution
93
Starch indicator
49
Screened methyl
orange
Sea water
46
104
Silicate gels
86
Silver electroplating
36
Silver nitrate(V)
77
91
92
Sterilising conditions
Strontium choride
95
89
Sugar chromatography
26
100
49
100
78
Soap
79
Sulfur dioxide
preparation
Sulfur dioxide test
96
Slime
Sodium carbonate
80
97
94
Sulfuric(VI) acid
98
Technical agar
Sodium carbonate
(standard solution)
Sodium chlorate(I)
81
Sodium chloride
82
Sodium dihydrogen
phosphate(V)
67
Sodium dihydrogen
phosphate(V) -1- water
67
49
CLEAPSS 2011
123
99
104
99
100
101
Thermochromic liquid
30
Thiooxamide
101
2,3,5-
Name
Recipe
Sheet
Recipe
Sheet
29
Unsaturation in organic
chemistry
102
49
Vitamin C test
13
79
Washing soda
80
Water
104
66
105
Yamada indicator
47
Zinc electroplating
36
Zinc sulphate
106
Thymol blue
46
Thymolphthalein
46
Tin(II) chloride
Tollens reagent
103
102
Triphenyl tetrazolium
chloride
Trypsin
13
TTC
13
Universal buffer
18
Universal indicator
47
37
124
CLEAPSS 2011