RIBBONS OF BLUE/ WATERWATCH WA

WATER CHEMISTRY SERIES

The chemistry behind water quality testing

Prepared by Stephanie Degens

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Editor/compiler Stephanie Degens. Community Monitoring and Environmental Officer Ribbons of Blue/Waterwatch WA Laboratory exercises author Allan Knight Acknowledgments: Ribbons of Blue/Waterwatch WA Regional Coordinators and their TEE chemistry teachers: For reading draft documents and trialing them with students. Staff from the Water and Rivers Commission. For comments on the draft documents. Water and Rivers Commission, Swan-Canning Cleanup Program and Swan River Trust: For support of the Ribbons of Blue/Waterwatch WA program.

For more information contact: State Facilitator Ribbons of Blue/Waterwatch WA on 9278 0300 or visit our web site www.wrc.wa.gov.au/ribbons ISBN: 0-7309-7593-2

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CONTENTS
Electrical Conductivity of Aqueous Solutions
Electrical Conductivity in Aquatic Systems Measuring Conductivity Background Experimental Processing of results, and questions 1 3 5 5

Dissolved Oxygen
Oxygen in Aquatic Systems The analysis of Dissolved Oxygen in Water Background Experimental Preparation of primary potassium bi-iodate solution Preparation and standardisation of a sodium thiosulfate solution Processing of results, and questions Analysis of dissolved oxygen in a natural water sample Processing of results, and questions Biological Oxygen Demand Considerations 7 9 10 11 12 13 14 14 15

pH and Aquatic Systems

pH Buffering of Aquatic Systems Factors Affecting pH of Aqueous Systems pH Measurement Background Experimental Processing of results, and questions 17 18 19 20 20

Plant Nutrients - Nitrogen

Nitrogen in water Nitrogen cycle Analysis of nitrate/nitrite in a natural water sample Background Experimental Processing of results, and questions 23 23 26 28 28

Plant Nutrients Phosphorus

Phosphorus in water Analysis of phosphate in a natural water sample Background Experimental Processing of results, and questions 30 32 34 34

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ELECTRICAL CONDUCTIVITY OF AQUEOUS SOLUTIONS

TEE Year
11

12

Subject Objectives from Chemistry Syllabus (2000-2001) 1.38, 1.49, 1.L.6, 7.15 1.2, 2.L.1,

Electrical Conductivity in Aquatic Systems

The electrical conductivity of aqueous solutions is dependent upon the ion concentration of the solution. Ions are free to move within the body of water and thus are capable of carrying a current. The greater the ion concentration the higher the conductivity. Note: Conductivity also increases with temperature. Conductivity is used as a measure of salinity, the salt content of the water. Sodium chloride is the main contributor to water salinity but other salts present may include calcium carbonate (limestone) and other calcium and magnesium salts. The conductivity/salinity can fluctuate naturally, especially in estuarine systems, however it can be abnormally increased by human activity in a range of ways. Overuse of fertilisers, causing leaching from the soil, can lead to an increase in the concentration of phosphate, nitrate and ammonium ions (and their associated cations/anions) thus leading to increased conductivity. Excessive removal of native deep-rooted vegetation is of major concern in Western Australia (as well as other parts of Australia). Removal of vegetation has caused a rise in the level of underground water tables in many areas. As the water table rises it dissolves salt that was previously held in the soil and brings it closer to the surface. If the watertable rises to such an extent that it comes into contact with surface water the salt can eventually enter rivers and streams and increase their salinity. That is reduce the freshness of the water. This is termed as dryland salinity. This can have adverse effects on the organisms depending upon this water for their well-being. The rising water table can also cause an increase in bore water salinity. The salinity of the Swan-Canning River system varies from fresh-to-brackish conditions in the winter and spring to salty during the summer and autumn. When river flow declines at the end of winter, sea water moves progressively up the estuary reaching the upper Swan Maylands area and the Kent Street Weir in spring to early summer. (In low rainfall years significant quantities of salty water often remains in the system over winter.) The salt water flows in as a wedge beneath the less dense fresh water (see figure 1). This layering process is called stratification. The difference in densities reduces mixing of surface and bottom waters thus preventing oxygen replenishment of the bottom waters. The oxygen concentration in the bottom waters is further lowered as decomposition of organic matter consumes remaining oxygen. (This then causes the release of nutrients, such as nitrate and

phosphate, stored in the sediments. This occurs because the solubilities of these parameters tend to be higher in water with low oxygen concentration. These dissolved nutrients accumulate in the stagnant salty waters at the bottom.) Figure 1. Salt wedge in the Swan River (Swan-Canning Cleanup Program Action Plan, 1999 11)

Every spring a dense wedge of salty water moves upstream while lighter fresh water flows over the top as the summer progresses. The wedge moves upstream according to tidal and barometric influences, and freshwater flows reduce. The leading edge of the salt wedge often has very low oxygen and high nutrient levels. (Swan-Canning Cleanup Program Action Plan, 1999 11)

Measuring Conductivity
Background Conductivity can be measured relatively accurately with a conductivity meter. The instruments measure the amount of electrical charge passing between two metal electrodes 1 cm apart. The units of salinity/conductivity can be expressed as microsiemens per centimetre (S/cm), millisiemens per centimetre (mS/cm) or millisiemens per metre (mS/m). Be aware of the units in which your instrument displays its readings. The total soluble salts, expressed in milligrams per litre (mg/L), can be calculated from the following standard conversion between different units: Electrical conductivity (S/cm) 0.55 = total soluble salts (mg/L) Electrical conductivity (mS/cm) 550 = total soluble salts (mg/L) Electrical conductivity (mS/m) 5.5 = total soluble salts (mg/L) As conductivity increases with temperature, it is necessary to compensate for temperature variations between measurements. Table 1 shows ranges of water electrical conductivity (EC) and restrictions that EC places on water use.

Table 1. Electrical conductivity ranges

Salty

Over 10 000 S/cm or over 5 500 mg/L

Very Brackish To Salty

2 500 10 000 S/cm or 1 375 5 500 mg/L

Moderate Brackish

800 2 500 S/cm or 440 1 375 mg/L

Fresh

Electrical conductivity (EC) 0 - 800 S/cm or 0 - 440 mg/L

Characteristics of the water EC ranges, restrictions for usage and specific ECs.
Good drinking water for humans. Generally good for irrigation, though above 300 S/cm some care must be taken if using overhead spraying of salt sensitive plants. Suitable for all livestock. Distilled water 0 S/cm. Rain water 66 S/cm. Can be consumed by humans, though most would prefer water in the lower half of this range. When used for irrigation it requires special management including suitable soils, good drainage and consideration of salt tolerance of plants. Suitable for all livestock. Tap water 400 S/cm. Maximum for hot water systems 1 600 S/cm. Maximum for human drinking water 2 500 S/cm. Not recommended for human consumption although water of up to 3 000 S/cm could be drunk if nothing else was available. Not normally suitable for irrigation, though water of up to 6 000 S/cm can be used on very salt tolerant crops with special management. Over 6 000 S/cm, occasional emergency irrigation may be possible with care. When used for drinking water by poultry and pigs, the electrical conductivity should be limited to about 6 000 S/cm. Most other livestock can drink up to 10 000 S/cm. Not suitable for human consumption or irrigation. Not suitable for poultry, pigs or lactating animals, but beef cattle can use water up to 17 000 S/cm and adult sheep on dry feed can tolerate 23 000 S/cm. However chemical analysis should be considered before using high electrical conductivity water for stock. Water up to 50 000 S/cm (the salinity of the sea) can be used (i) to flush toilets provided corrosion in the cistern can be controlled and (ii) for making concrete, provided the reinforcement is well covered. Pacific Ocean 58 000 S/cm. Dead Sea 550 000 S/cm.

Experimental
Equipment Water sample(s) Thermometer (or temperature probe) Conductivity meter Procedure Note: The conductivity can be measured in the field or water samples collected and measurements made in the laboratory. 1. Your Ribbons of Blue/Waterwatch WA Regional Coordinator will support you with information on water sampling techniques and help you to use the method appropriate for your water-body to collect a sample(s). 2. Calibrate your conductivity meter as described in the instructions supplied with your meter or by your Regional Coordinator. 3. Test and record the EC of the Mystery Solution supplied by Ribbons of Blue, to ensure the equipment is working accurately. 4. Measure and record the conductivity of the water sample(s). Processing of results, and questions 1. Calculate the total soluble salts (mg/L) using the conversion factors provided on page 3. 2. Assess the water quality based on salinity using the information provided in Table 1. 3. Suggest the ion species most likely to be responsible for the electrical conductivity of your water sample. 4. At what time of year would you expect the salinity of your waterbody to be lowest? Why?

DISSOLVED OXYGEN
TEE Year 11 12 Subject Objectives from Chemistry Syllabus (2000-2001) 1.44, 2.48, 2.60, 2.61, 2.62, 2.63, 2.72, 2.73, 5.2, 5.5, 5.6, 5.7, 5.35, 5.36, 5.37, 5.39,

Oxygen in Aquatic Systems

The solubility of gases in liquids can be studied in the context of oxygen in natural and man made waterbodies (lakes, rivers, streams, drains etc.). Factors affecting the solubility of oxygen in water include: Temperature increasing solubility with decreasing temperature Atmospheric pressure (altitude) the greater the pressure the higher the solubility Salt concentration the lower the salt concentration the higher the oxygen concentration The sources and consumption of oxygen in water bodies also influence the amount of dissolved oxygen. The sources are: Absorption There is continuous exchange of oxygen between water and the surrounding air. The greater the contact between the water and the air the more oxygen that can dissolve, thus a turbulent stream will tend to have a higher oxygen concentration than a still body of water. Photosynthesis This redox process carried out by aquatic (and land) plants results in oxygen directly entering the water. Those things that reduce the amount of sunlight able to penetrate the water (e.g. suspended solids) will lower the rate of photosynthesis and hence lower oxygen concentration. As photosynthesis takes place only during the day, the concentration of dissolved oxygen will vary over the 24-hour daily cycle. Levels peak early afternoon and are lowest just before sunrise. Oxygen is consumed by: Respiration all organisms (aquatic or terrestrial) consume oxygen during respiration. Decomposition the decomposition of plant and animal waste (whether from living or dead organisms) is carried out by bacteria and other micro-organisms that use oxygen to oxidise the organic matter. The level of organic wastes present in a water sample can be estimated by measuring oxygen consumption in the water. The biological oxygen demand (BOD) provides a measure of the amount of oxygen consumed by a sample of water under standard conditions. The oxygen concentration in the water is measured immediately on collection and again after the sample has been incubated at 20C for 5 days. The difference in oxygen concentrations is given in units of milligrams per litre (mg/L) or parts per million (ppm) and is an indication of the quantity of organic wastes in the sample.

The low solubility of oxygen in water can be explained by the nature of the solvent and solute. Water has a polar molecule whilst the oxygen molecule is non-polar. The predominant type of molecular interactions between water molecules is hydrogen bonding and between oxygen molecules are dispersion forces. Thus there will be little interaction between water molecules and oxygen molecules, and so the solubility of oxygen in water is low. Oxygen is essential for the survival of aquatic organisms, and a shortage of dissolved oxygen is not only a sign of pollution, it is harmful to fish. The sensitivity of aquatic species to oxygen depletion varies, but some general guidelines to consider when analysing test results are: 5 6 ppm Sufficient for most species < 4 ppm Stressful to most aquatic species < 2 ppm Fatal to most species Work by the Waters and Rivers Commission to increase the oxygen concentration in the Canning River has included an oxygenation technique involving the injection of oxygen rich water into the colder deeper water (see Figure 2). Figure 2. Oxygenation plant on the Canning River (Swan Canning Cleanup Program Action Plan, 1999 53)

The purpose of oxygenation is to pump low-oxygen water from the bottom of the river to a mixing plant to increase oxygen content and then return the treated water. Oxygenation thus specifically targets the low-oxygen layer of the river and does not interrupt natural layering caused by salinity and temperature differences. BOC gases donated material and expertise for a trial conducted on the Canning River in 199798. In the Thames River (London), authorities utilise a barge that injects oxygen directly to the bottom of the river to solve the problem of poor water quality following storm and pollution events. (Swan Canning Cleanup Program Action Plan, 1999 53)

The analysis of dissolved oxygen in water

Background The analysis of natural water samples for dissolved oxygen can be done using a redox titration known as the Winkler method. In this procedure, the water sample is first treated with an excess of manganese II, potassium iodide and sodium hydroxide. The white manganese II hydroxide that initially precipitates is oxidised readily by dissolved oxygen to give the brown manganese III hydroxide. The reactions are Mn2+(aq) + 2OH-(aq) Mn(OH)2(S) 4Mn(OH)2(S) + O2 + 2H2O 4Mn(OH)3(S) When acidified, the manganese III hydroxide dissolves and the freed manganese III ion oxidises iodide to iodine. Mn(OH)3(S) + 3H+(aq) Mn3+(aq) + 3H2O(l) 2Mn3+(aq) + 2I- I2 + 2Mn2+(aq) The liberated iodine can then be titrated with thiosulfate (S2O32-). 2 S2O32-(aq) + I2(aq) S4O62-(aq) + 2I-(aq) The sequence of laboratory activities involved in the Winkler analysis is as follows: I. Preparation of potassium bi-iodate solution as a primary standard II. Preparation and standardisation of a sodium thiosulfate solution III. Analysis of the dissolved oxygen in a natural water sample Table 2 gives an indication of what your dissolved oxygen results mean for the waterbody sampled. Table 2. Rating of dissolved oxygen ranges in natural waterbodies DISSOLVED OXYGEN (percent saturation) in standing or flowing water Normal Some High Pollution Pollution 80 120 120 135 > 135 or or 55 80 < 55

Experimental
Preparation of primary potassium bi-iodate solution Equipment Drying oven Beaker (250 mL) Desiccator Balance Volumetric flask (1.00 L) Wash bottle Storage bottle (1.00 L, dark glass) Deionised water Primary standard potassium bi-iodate [KH(IO3)2] Procedure 1. Calculate the mass of anhydrous KH(IO3)2 needed to make up 1.00 L of approximately 8.35 10-4 mol/L solution. 2. Place a little more than this calculated mass in an oven at 105 C for 1 hour. After drying place the KH(IO3)2 in a desiccator to cool. 3. Accurately weigh out into a beaker a mass of KH(IO3)2 approximately equal to that calculated in step 1. 4. Dissolve the solid in about 100 mL of water and then transfer the solution to the volumetric flask and make up to the graduated mark. Stopper the flask and mix the solution thoroughly. 5. Transfer the solution to the clean dark glass storage bottle and store in a dark cupboard until required.

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Preparation and standardisation of a sodium thiosulfate solution Equipment Bottle (250 mL) Beaker (250 mL) Volumetric flask (1.00 L) Conical flask (250 mL) Graduated cylinders (10 mL and 100 mL) Burette and stand Funnel Pipette (10 mL) Pipette filler Deionized water Alkaline-iodide-azide reagent (1.0 mL) Concentrated sulfuric acid [H2SO4] (1.0 mL) Starch indicator solution (usually VITEX in deionized water) Sodium thiosulfate pentahydrate [Na2S2O3.5H2O] (~5 g) Sodium carbonate [Na2CO3] (0.1 g) Manganese II sulfate solution (1.0 mL) Alkaline-iodide-azide Reagent The alkaline-iodide-azide reagent is prepared as follows (probably best done by the laboratory technician prior to the laboratory activity): 1. Dissolve, in small increments, 400 g of sodium hydroxide pellets in 500 mL of freshly boiled deionized water. 2. Add 900 g of sodium iodide while the solution is still hot. 3. Dissolve 10 g of sodium azide, NaN3, in 40 mL of deionized water. 4. Add the sodium azide solution to the first solution (step 2) and make up to 1.0 L. Manganese II sulfate solution The manganese II sulfate solution is prepared as follows (probably best done by the laboratory technician prior to the laboratory activity): 1. Dissolve 365 g of manganese II sulfate monohydrate, MnSO4.H2O, in freshly boiled deionized water and dilute to 1.00 L. Note: Manganese II sulfate dihydrate (400 g) or tetrahydrate (480 g) can be used. Procedure 1. Dissolve approximately 4.8 g of Na2S2O3.5H2O and 0.1 g of Na2CO3 in about 100 mL of deionized water in a beaker. 2. Transfer to a 1.00 L volumetric flask and make up to the graduated mark. Mix the solution thoroughly.

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3. Fill a 250 mL bottle with deionized water (NOT chlorinated tap water). To this add, mixing after each addition, 1.0 mL alkaline-iodide-azide reagent, 1.0 mL concentrated sulfuric acid, and 1.0 mL manganese II sulfate solution. 4. Transfer 100 mL of this solution to a conical flask and pipette a 10 mL aliquot of the potassium bi-iodate solution into the flask. 5. Allow the mixture to stand for 2 minutes in the dark and then titrate the liberated iodine with thiosulfate solution until a very pale straw colour develops in the solution. Add four drops of starch indicator solution and continue titrating until the blue colour just disappears. Processing of results, and questions 1. In the preparations of the alkaline-iodide-azide reagent and manganese II sulfate solution why is it necessary to use boiled deionized water? 2. From the volume of thiosulfate solution used, calculate its concentration. 3. Why is it not acceptable to use the sodium thiosulfate as a primary standard?

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Analysis of dissolved oxygen in a natural water sample

Equipment Biological Oxygen Demand Bottle or ordinary glass stoppered bottle (250 mL) Conical flask (250 mL) Graduated cylinder (10 mL) Burette and stand Funnel Pipette (20 mL) Pipette filler Deionized water Manganese II sulfate solution (1.0 mL) Alkaline-iodide-azide reagent (1.0 mL) Concentrated sulfuric acid [H2SO4] (1.0 mL) Sodium thiosulfate pentahydrate [Na2S2O3.5H2O] (~5 g) Starch indicator solution (usually VITEX in deionized water)

Note: If you wish to carry out a BOD measurement collect sufficient water to obtain two sets of results 5 days apart. Procedure Note: When adding solutions to the water sample place all pipettes below the water surface to avoid agitation. 1. Collect your water sample(s). To avoid contamination, thoroughly rinse the collection bottle with sample water. It is important to avoid trapping air bubbles in the bottle. Tightly cap the bottle and submerge to the desired depth. Remove the cap to fill the bottle to the top and then stopper or seal it whilst still submerged. Once the sample is collected the bottle should only be opened when the sample is to be analysed. 2. To the collected natural water sample, add 1.0 mL of manganese II sulfate solution followed immediately by 1.0 mL of alkaline-iodide-azide reagent. (Some overflow of water will occur.) Re-stopper the bottle at once, ensure no air is trapped, and shake vigorously for at least 20 seconds or until the precipitated manganese II and manganese III hydroxide is evenly distributed. 3. Let the bottle stand for 2 3 minutes and then shake again. 4. Allow the precipitate in the bottle to settle by at least 1/3 (10 20 minutes). 5. Add 1.0 mL of concentrated, reagent grade H2SO4 well below the surface. (The overflow will be alkaline so avoid contact with your skin.) Re-stopper the flask and shake gently until dissolution of the precipitate. 6. Using a pipette, transfer 100 mL of the solution to a conical flask.

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7. Titrate at once with the previously standardised thiosulfate solution until a very pale straw colour appears. 8. Add 4 drops of starch indicator solution, and continue the titration until the blue colour just disappears. Processing of results, and questions 1. When collecting the water sample why is it important to ensure that no air is trapped in the bottle? 2. Which ion species in this process reacts directly with any oxygen dissolved in the water? 3. Suggest a reason why it is necessary to shake the bottle in step 1 of the procedure and then allow it to stand for 2 3 minutes as stated in step 2? 4. Calculate the number of moles of thiosulfate used in the titration? 5. What is the mole relationship between the thiosulfate and the oxygen? 6. How many moles of oxygen are present in 100 mL of your water sample? 7. What is the concentration of your dissolved oxygen in mg/L and ppm? 8. What volume of oxygen would this be at S.T.P.? 9. Rate the health of the water source from which your sample was taken. 10. Why would the accuracy of this technique for measuring dissolved oxygen be reduced by the presence of iron II (Fe2+) in the water sample?

Biological Oxygen Demand (BOD)

To obtain an indication of the amount of organic waste in your water sample by the BOD measurement, store your water sample at 20C for 5 days and then analyse for oxygen as described above. The difference between your two measurements is the 5 day BOD of your water. The oxygen consumed during this 5 day period has been used by bacteria and other micro-organisms in the oxidation of organic wastes in the water. Note: If you are unable to store the water sample at 20C, it can be kept at ambient temperature and the BOD measured for this temperature.

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Considerations This method is not applicable under the following conditions: 1. Samples containing more than 1 mg/L of iron II. This can be overcome by the addition of potassium fluoride solution prior to acidification. Potassium fluoride solution is made by dissolving 40g of KF.2H2O in distilled water and diluting to 100 mL. 2. Samples containing sulfite, thiosulfite, free chlorine or hypochlorite. 3. Samples with high concentrations of suspended solids. Filtration can remedy this. 4. Samples containing other oxidising or reducing ion species.

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pH AND AQUATIC SYSTEMS

TEE Year 11 12 Subject Objectives from Chemistry Syllabus (2000-2001) 4.1, 4.3, 4.4, 4.5, 4.12, 4.17, 3.17, 3.18, 3.21, 3.L.2, 4.4, 4.5, 4.6, 4.9, 4.13, 4.19.

pH Buffering of Aquatic Systems

The pH of natural fresh waters is usually about 7 but values may vary from ~5.0 8.5 depending upon factors such as soil type, geology and vegetation. Marine waters have a pH close to 8.2. Control of pH in many waters is achieved by the carbonate hydrogencarbonate buffer system. A buffer solution is one that maintains the approximate pH of a solution when a small amount of an acid or a base is added to the solution. A buffer solutions function is an application of the equilibrium principles as expressed in Le Chateliers Principle. The buffering of natural waters by the carbonate hydrogencarbonate system initially involves the dissolution of atmospheric carbon dioxide (or CO2 released by aquatic organisms into the water) to give carbonic acid. The reaction for this can be represented as H2O(l) + CO2(aq) H2CO3(aq)

The carbonic acid will ionize to a small extent to give hydronium and hydrogencarbonate ions: H2O(l) + H2CO3(aq) H3O+(aq) + HCO3-(aq)

The hydrogencarbonate ion will then hydrolyse as represented below: H2O(l) + HCO3-(aq) H3O+(aq) + CO32-(aq)

Addition of acid to the water results in an increase in hydronium ion concentration and as predicted by Le Chateliers principle the reaction favours the reverse direction. Conversely, the addition of a base would result in a reduction of hydronium ion concentration and so the reaction will shift to the products.

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Factors Affecting pH of Aqueous Systems

Many manufacturing and chemical industries release waste water into ground or surface waters in the environment at some stage of their process. The pH of this wastewater should, ideally, be the same as the pH of the natural water system into which it is being released. The presence of certain salts can influence the pH of aqueous systems. Some of these are present naturally in water but others are pollutants. Salts that are naturally present can have an unwanted effect on pH if their concentrations are increased by the actions of people. Hydrolysis of ionic compounds to give either hydronium ions or hydroxide ions can alter the normal pH of a natural aquatic system. People using inorganic phosphate fertilisers or the release of phosphate containing detergents can increase phosphate concentrations in waterbodies near to farming or residential areas. The hydrolysis of phosphate ion can increase the pH of the water as illustrated below: H2O(l) + PO43-(aq) HPO42-(aq) + OH-(aq)

The pH can also be increased by the presence of ammonia. H2O(l) + NH3(aq) NH4+(aq) + OH-(aq)

The application of ammonium based fertilisers can lead to a decrease in pH due to hydrolysis of the ammonium ion. H2O(l) + NH4+(aq) NH3(aq) + H3O+(aq)

Table 3 gives an indication of what your pH results mean for the waterbody sampled. Table 3. Rating of pH ranges for natural waterbodies pH in standing or flowing water Normal May be Polluted 5.0 7.0 (no 8.5 9.0 limestone) or 4.0 5.0 7.0-8.5 (limestone)

Pollution Problem <4 or >9

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pH Measurement
Background The pH of an aqueous solution can be measured with the use of pH paper, acid-base indicators, such as universal indicator, or pH meters or scans. In this activity, the pH of a natural water sample will be measured using a pH meter. When using a pH meter it is first necessary to calibrate the meter. Calibration involves placing the pH probe into a buffer solution of known pH to ensure the meter is reading values accurately. Conventionally two buffer solutions are chosen such that their pH values span the pH range of interest. For example, if the test sample were thought to have a pH of about 6, typical buffer solutions used for calibration would have values of, say, pH 4 and 9. The pH meter would be placed in the pH 4 buffer solution and, if necessary, the reading on the meter would be adjusted to 4. The pH probe would then be similarly placed in pH 9 buffer solution and the reading adjusted if required. A typical pH 4 buffer is a 0.05 mol/L solution of potassium hydrogenphthalate [KHO2CC6H4CO2 ] (A salt of an aromatic carboxylic acid. See Figure 3 below.) A typical pH 9 buffer is sodium tetraborate (borax) [Na2B4O7.10H20]. O C OH C OK O Figure 3: Potassium hydrogenphthalate

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Experimental
Equipment pH meter pH buffer solutions (pH 4 and 9) 2 beakers (100 mL) bottle (250 mL) water samples from your local environment Procedure Note: Remember to rinse the pH electrodes with distilled water before each measurement. 1. Calibrate your pH meter as follows: I. Rinse a clean beaker with a small quantity of the pH 4 buffer solution. II. Pour sufficient pH 4 buffer solution into the beaker to cover the glass electrode of the meter to the correct depth. (Your teacher will show you this.) III. Rinse the pH probe with pH 4 buffer solution and then place the electrode into the buffer solution and check the reading on the meter. If necessary adjust the reading to a value of 4. IV. Repeat steps I III for the pH 9 buffer solution. 2. Rinse a clean beaker with a small quantity of your water sample and then pour in sufficient quantity of your water sample to cover the glass electrode to the correct depth. Rinse the pH probe with sample water and then place it into the water, measure and record the pH value. 3. Repeat step 2 until you have measured the pH for all your water samples. Processing of results, and questions 1. Write the hydrolysis equation for the phthalate ion, HO2CC6H4CO2-, in the pH 4 buffer solution (Hint: Remember the solution is acidic.) Using the appropriate chemical principles, explain why the addition of a small quantity of acid or base to this solution should not greatly alter its pH. 2. Write the hydrolysis equation for the tetraborate ion, B4O72-, from the pH 9 solution. Using the appropriate chemical principles, explain why the addition of a small quantity of acid or base to this solution should not greatly alter its pH. Note: The following questions can probably only be answered if students have access to any long term results of pH measurements made of waterbodies in their local environment or they have made measurements of phosphate, ammonia or ammonium levels in the water samples.

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3. Try to account for any variations in the pH of your water samples. Use appropriate equations to explain any of your results. 4. Assess the quality of your water sample based on pH using the information provided in Table 3.

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PLANT NUTRIENTS - NITROGEN

TEE Year 11 12 Subject Objectives from Chemistry Syllabus (2000-2001) 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.1, 5.2, 5.3, 5.5, 5.6, 5.7, 6.5, 6.14

Nitrogen in water
Nitrogen is a part of living organisms in their protein, DNA and other components. It is released naturally into water with the decay of dead organisms and their wastes and through the fixation of atmospheric nitrogen. Humans introduce nitrogen into waterbodies through a number of sources including: fertilisers plant and animal wastes rural and urban run-off sewage effluent industrial discharges The total nitrogen present in the water at any given time is composed of organic nitrogen and inorganic nitrogen. Organic nitrogen is often associated with biological material and is not soluble and consequently is less mobile that inorganic nitrogen. Inorganic nitrogen is made up of dissolved or particulate nitrate, nitrite and ammonium. The majority of nitrate occurs in dissolved form due to the high solubility of nitrate compounds. This contrasts to the phosphates that have low solubility and so often occur in a particulate form.

Nitrogen cycle
A large amount of nitrogen enters the Swan-Canning system in dissolved form. This provides nutrients for spring algal blooms that take up the nitrate and convert it to organic nitrogen as algal cells. After spring these algae die and fall to the sediment on the river floor. In the sediment, decomposition by microbes converts the nitrogen to ammonia and then nitrate. This ammonia and nitrate is now available for uptake by more algae in the summer, which subsequently die, fall to the bottom and decompose. This cycle continues throughout summer.

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Figure 4. Nitrogen cycle in a freshwater system

Biodegradation of organic nitrogen compounds produces ammonium ion as the first inorganic species. Nitrification is a two-stage redox process in which microorganisms convert ammonium ion to nitrate ion. In the first stage the ammonium is oxidised to nitrite ion. 2NH4+(aq) + 3O2(g) 2NO2-(aq) + 4H+(aq) + 2H2O(l) The nitrite ion is then oxidised to nitrate: 2NO2-(aq) + O2(g) 2NO3-(aq) This process uses considerable quantities of oxygen and contributes significantly to reducing the amount of oxygen available to other aquatic organisms. Nitrate concentration can also increase in summer. The incoming salt wedge reduces the amount of dissolved oxygen in bottom waters. Microbes release nitrogen compounds under these conditions.

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Nitrogen can only be lost from a water system in the following ways: denitrification, - where nitrate is reduced to N2 which then escapes into the atmosphere uptake by living organisms burial in sediments export to the ocean Denitrification is when bacterial enzymes catalyse the reduction of nitrate to nitrogen gas by a reducing agent such as carbohydrate (Sugars, such as glucose [C6H12O6], are an example of carbohydrates.): 24NO3-(aq) + 5 C6H12O6(aq) + 24H+(aq) 12N2(g) + 30CO2(g) + 42H2O(l) The nitrogen gas can then escape to the atmosphere. The processes of nitrification and denitrification as well as movement of nitrogen through a freshwater system are illustrated in Figure 4. Nitrogen is also found in groundwater. It percolates through the soil and collects in aquifers, most commonly as nitrate. Some stream systems and wetlands are groundwater fed. Groundwater nitrogen can therefore enter the surface water system. Table 4 gives an indication of what your nitrate results mean for the waterbody sampled.

Table 4. Rating of nitrate content in waterbodies Water Type

STANDING FLOWING TANNIN

Nitrate Content in milligrams per litre (mg/L) Low Medium High 0 0.025 0.025 0.25 > 0.25 0 0.05 0 0.25 0.05 0.4 0.25 1.5 > 0.4 > 1.5

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Analysis of nitrate/nitrite in a natural water sample

Background The free (dissolved) nitrate tests used in water testing kits today are based on a method developed in the late 1800s. The concentration of free nitrate in a water sample can be determined via a colorimetric technique. This technique involves converting colourless nitrate to a pink coloured compound by reacting a series of chemicals with a filtered water sample. The intensity of colour produced is directly proportional to the amount of free nitrate in the water sample. The colour of the tested water sample can be compared to a set of known colour standards and thus the concentration of the nitrate estimated. Alternatively, the amount of nitrate in a sample can be determine using a photometer and comparing light transmittance to a calibration chart. Free nitrate tests should be carried out on the filtrate from water samples passed through 0.45 m filter paper. However, due to costs, Ribbons of Blue uses GFC filter papers which filter out particles greater than 1.2 m Note: The test described below actually measures both the nitrate and nitrite in the sample but the nitrite is usually at such a low concentration that the results are fairly accurate. The combination of the nitrate and nitrite is known as the total oxidised nitrogen (TON). The first step in the process is the reduction of nitrate ion to nitrite by the addition of cadmium or zinc to the water sample. Below is the reaction for cadmium with nitrate. NO3-(aq) + Cd(s) + 2H+(aq) NO2-(aq) + Cd2+(aq) + H2O(l)
Palintest Kit uses Nitratest Powder containing 70% zinc as the reducing agent. The LaMotte Kit uses Nitrate Reducing Agent that contains 7% Cadmium, which is added after the Mixed Acid Reagent.

The nitrite produced thus is determined by a diazotisation reaction. In acid solution, nitrite ions and aromatic amines react to form diazonium salts. HO3S NH2 + NO2 + 2H+ HO3S N2+ + 2H2O

aromatic amine

diazonium salt

In the Palintest Kit, sulfanilic acid, contained in the Nitricol Tablets acts as the aromatic amine. The acid solution is provided by the ammonium chloride, from the Nitratest Tablets, which dissociates to form a weak acid, and the sulfanilic acid. The LaMotte Kit method has already acidified the water sample by adding the Mixed Acid Reagent containing Citric and Acetic Acid. The aromatic amine is sulfanilamide contained in the Nitrate Reducing Powder.

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The diazonium salts can then in turn react with diaromatic amines to form intensely coloured azo compounds. (Aromatic compounds are those that contain a benzene ring and amines are characterised by the functional group NH2.) For example: HO3S N2++ NH2R HO3S N2 NH2R + H+

diaromatic amine

aromatic azo compound (highly colouredred in this case)

The Palintest Kit uses the diaromatic amine N-1-napthlethylene diamine dihydrochloride contained in the Nitricol Tablet to form the azo-compound. The LaMotte Kit also uses the diaromatic amine N-1-napthlethylene diamine dihydrochloride contained in the Nitrate Reducing Agent to form the azo-compound.

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Experimental
Equipment Water sample(s) Palintest Kit or other Nitrate testing kit Procedure Note: If you wish to measure the nitrite and nitrate concentrations separately you can do so by omitting the addition of the nitrate reducing reagent to a sample of water. Add only the second reagent to give the colour. An estimate of the nitrite concentration can then be made by obtaining a percent transmission reading and reading from the table of values. The nitrate concentration can then be obtained by carrying out the full test as outlined in the test kit and subtracting the nitrite concentration from this second value. 1. Collect your water sample(s). Be sure to rinse your bottles with the water to be collected before obtaining your sample. Record the site(s) from which your sample is taken. If possible also record the depth and temperature from which you obtained your water. 2. Read the instructions in the test kit to determine the nitrate content of your water sample. If possible test a number of samples at different sites of the waterbody (this may be done as a class) to ascertain a profile of the waterbody. Processing of results, and questions 1. Using the outlined procedure in your kit, state the concentration of the nitrate in your water sample(s). Rate the concentration(s) using information from Table 4. 2. Suggest likely sources of nitrate for the waterbody you have tested. 3. What is the oxidation state of the nitrogen before and after reaction with the zinc (or cadmium)? (The Palintest uses zinc as the nitrate reducing agent.) 4. Write a balanced redox equation for the reaction between zinc and nitrate ion. 5. One of the ingredients used in the reagent tablets for this test is an acid. Suggest a reason for its inclusion? 6. Based on the value for the nitrate concentration you determined, calculate the mass of zinc (or cadmium, as appropriate to the test kit used) needed to react with this quantity of nitrate. Is the quantity of zinc (or cadmium) added to your water sample likely to be more or less than your calculated value? Why? 7. In the nitrification process, ammonium ion is converted to nitrite ion, which in turn is converted to nitrate. What are the oxidation states of the nitrogen in these three species? Explain why it is possible for nitrogen to have this range of oxidation states. (Hint: Consider the electron configuration of nitrogen.)

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8. In the denitrification process, nitrate is converted to nitrogen gas. Write a balanced redox equation for this conversion where sucrose, C12H22O11, is the reducing agent. (Assume the sucrose is oxidised to carbon dioxide.) 9. Why does most of the nitrogen gas produced in denitrification escape to the atmosphere rather than dissolve in the water?

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PLANT NUTRIENTS - PHOSPHORUS

TEE Year 11 12 Subject Objectives from Chemistry Syllabus (2000-2001) 1.36, 1.49, 5.2, 5.3, 5.4, 5.5, 5.6 3.19, 3.20, 3.21, 3.L.2, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 7.5, 7.6, 7.7, 7.14

Phosphorus in water
The total phosphorus content of a waterbody consists of three components. Phosphorus can occur in the form of phosphate bound up in the organic matter (organic phosphate), as dissolved phosphate free in the water (known as reactive phosphate) or as particulate phosphate bound to clay or insoluble metal phosphate compounds (particularly iron and aluminium oxides). Phosphorus is one of the principal plant nutrients entering the Swan-Canning river system. In most natural systems phosphorus is present as either organic or inorganic phosphate. As no simple method exists for measuring phosphorus content, and it is rare in its pure form, the following method is for analysis of dissolved phosphate. This procedure will identify the amount of phosphorus in the water sample that exists as phosphate ions (PO43-). Compounds such as calcium phosphate (Ca3(PO4)2) will have dissociated in the sample after the addition of acid so this phosphate will also be detected. We commonly work out phosphorus content by working out how much phosphate occurs in a sample and then calculating how much of this is phosphorus (since the phosphorus content of phosphate is always constant). Natural sources of phosphate in an undisturbed system include weathering of rocks and decomposition of organic matter. These natural sources lead to a low phosphate concentration in the water. Human activity significantly increases the concentration of phosphate. Human activities that cause phosphate to enter waterbodies include: application of inorganic fertilisers, both on home gardens and agricultural lands use of detergents organic wastes (ie. from garden, agricultural wastes and animal effluent) sewage effluent industrial discharges

The major problem that can arise from an excess of phosphates (and other plant nutrients) in water is eutrophication. This is the process whereby the excess plant nutrients cause an explosion in the algal population in a waterbody. When the algae die it is decomposed by micro-organisms which consume much of the oxygen from the water. This reduces the oxygen concentration to a level where fish and other aquatic organisms have insufficient for their survival.

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(Some of the algae species present can have adverse health effects on humans. The extent of these species is not a major threat in the Swan-Canning system as yet but there have been occasions during the summer when health authorities have advised against swimming in parts of the river where blooms have been present.) Phosphate applied as fertiliser is usually soluble and it is as a result of fertiliser use that most phosphate enters the Swan River. In most soils the phosphate is immobilised by reacting with iron, manganese, calcium and aluminium ions to form less soluble compounds (refer to Solubility Rules) or by being adsorbed onto the surfaces of clay and silt particles. It is then said to be bound and so much less mobile. Bound phosphate only enters waterbodies as part of soil carried into the waterbodies. However, the sandy soils of the Swan Coastal Plain contain few binding metals and little clay or silt. This results in a high concentration of dissolved phosphate entering ground and surface waters. Phosphate concentration is also influenced by the quantity of dissolved oxygen. Under oxygenated conditions, phosphorus can be retained in sediment by the formation of iron, manganese, aluminium and calcium compounds. The lower oxygen concentrations during summer caused by the salt wedge moving up the Swan River (and generally higher temperatures and reduced flow rates) can result in release of phosphate into the water column.

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Analysis of phosphate in a natural water sample

Background The measurement of total phosphate content in the water requires the conversion of the particulate forms into the dissolved reactive form. The conversion can be carried out by acid hydrolysis at boiling water temperature. Acid hydrolysis involves mixing concentrated acid with the water sample to react with the particulate forms of phosphorus. For example, if there were suspended particles of calcium phosphate in the water, reaction with concentrated acid would produce the soluble calcium hydrogenphosphate. This is an equilibrium process whereby the hydrogen ions initially react with free phosphate to give hydrogenphosphate: H+(aq) + PO43-(aq) HPO42-(aq)

This reaction results in a lowering of the free phosphate ion concentration thus units of the particulate phosphates, such as calcium phosphate, will dissolve: Ca3(PO4)2(s) 3Ca2+(aq) + 2PO43-(aq)

Note: When we say that an ionic compound is insoluble, as in the Solubility Rules, this is not strictly accurate. Those that we say are insoluble should more accurately be described as having very low solubility. For any ionic compound in water some dissociation will take place. An equilibrium is established between the solid and the dissociated ions. Hence by adding excess acid to an insoluble phosphate, the concentration of phosphate ions is reduced and the equilibrium is altered in such a way as to cause the solid to dissolve. It is not practical or safe to measure the total phosphate content in a water sample out in the field. Therefore, Ribbons of Blue/Waterwatch WA only analyses for dissolved phosphate. Acid hydrolysis is carried out at room temperature. The dissolved phosphate tests used to analysis water samples are based on the Ascorbic Acid Method. The concentration of dissolved phosphate in a water sample can be determined via a colorimetric technique. This technique involves converting colourless phosphate to a blue coloured compound by reacting a series of chemicals with a filtered water sample. The intensity of colour produced is directly proportional to the amount of dissolved phosphate in the water sample. The colour of the tested water sample can be compared to a set of known colour standards and thus the concentration of the phosphate estimated. Alternatively, the amount of phosphate in a sample can be determine using a photometer and comparing light transmittance to a calibration chart. Dissolved phosphate tests should be carried out on the filtrate from water samples passed through 0.45 m filter paper. However, due to costs, Ribbons of Blue/Waterwatch WA uses GFC filter papers which filter out particles greater than 1.2 m

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The two acid hydrolysis reactions can be represented in one step as: Ca3(PO4)2(s) + 2H+(aq) 3Ca2+(aq) + 2HPO42-(aq)
The Palintest Kit uses potassium hydrogen sulphate from the Palintest Phosphate No 1 LR tablet to acidify the water sample. The LaMotte Kit uses sulphuric acid from the Phosphate Acid Reagent to acidify the water sample.

The concentration of phosphate in a filtered water sample can be determined via a colorimetric technique. The colorimetric technique involves the reaction of the hydrogenphosphate ion, in the presence of acid, with ammonium molybdate, (NH4)6Mo7O24, to form ammonium molybdophosphate, (NH4)3[P(Mo3O10)4]. This is a complex reaction and may be represented as follows (This is not a balanced equation.) MoO42-(aq) + HPO42-(aq) [P(Mo3O10)4]3-(aq)
The Palintest Kit uses Ammonium Molybdate from the Palintest Phosphate No 2 LR tablet to facilitate the above reaction. The LaMotte Kit uses Ammonium Molybdate from the Phosphate Acid Reagent to facilitate the above reaction.

The molybdophosphate ion, [P(Mo3O10)4]3-, can then be reduced with a species such as acidified tin II or ascorbic acid (also known as vitamin C), molecular formula C6H8O6. This reduction reaction produces the intensely blue-coloured phosphorus molybdenum blue, (MoO2.4MoO3)2. H3PO4. The exhibiting of colour is a property typical of many transition metal compounds. The reduction using tin II is represented by the following equation: [P(Mo3O10)4]3- + 11H+ + 4Sn2+ (MoO2.4MoO3)2. H3PO4 + 2MoO2 + 4Sn4+ + 4H2O The intensity of the blue colour is proportional to the concentration of phosphate in the filtered water sample.
Palintest Kit reduces the molybdophosphate ion using sodium metabisulphate from the Palintest Phosphate No 2 LR tablet. LaMotte Kit reduces the molybdophosphate ion using Ascorbic acid from the Phosphate Reducing Agent.

Table 5 gives an indication of what your phosphate results mean for the waterbody sampled.

Table 5. Rating of phosphate content in waterbodies Water Type

STANDING FLOWING TANNIN

Phosphate Content in milligrams per litre (mg/L) Low Medium High 0 0.005 0.005 0.05 > 0.05 0 0.01 0 0.05 0.01 0.1 0.05 0.2 > 0.1 > 0.2

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Experimental
Equipment Water sample(s) Palintest Kit or other Phosphate testing kit Procedure Note: If you wish to test for reactive phosphate separately from total phosphorus your water sample must first be filtered. Standard chemical methods require a 0.45 m cellulose nitrate filter paper, however due to the expense of these filter papers Ribbons of Blue groups use 1.2 m GFC filter papers. 1. Collect your water sample(s). Be sure to rinse your bottles with the water to be collected before obtaining your sample. Record the site(s) from which your sample is taken. If possible also record the depth and temperature from which you obtained your water. 2. Read the instructions in the test kit on how to determine the phosphate content of your water sample. If possible test a number of samples at different sites of the water-body (this may be done as a class) to ascertain a profile of the waterbody. Processing of results, and questions 1. Using the outlined procedure in your kit, state the concentration of the phosphorus in your water sample(s). Rate the concentration(s) using information from Table 5. 2. Suggest likely sources of phosphate for the waterbody you have tested. 3. The formation of the blue colour depends upon a redox reaction. Give the oxidation states of the molybdenum before and after reaction with the hydrogenphosphate. Write a balanced half equation for the reduction of the molybdophosphate ion to the phosphorus molybdenum blue. 4. Assuming the ascorbic acid is oxidised completely to carbon dioxide write the half equation for its oxidation. Note: Some kits do not use ascorbic acid. 5. Combine the two half equations to give the redox equation for the reaction between the molybdophosphate ion and the ascorbic acid solution (or other reducing agent). 6. Which of the reagents in the reaction between the molybdophosphate ion and the ascorbic acid would you expect to be the limiting reagent? Explain why this is necessary to the success of the test. 7. Explain why most phosphate occurs in particulate form. 8. What effect(s) can the water temperature have on the concentration of phosphate? 9. How might the depth from which the water sample was taken influence the phosphate concentration?

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