ECP 419 PRACTICALS

1.0 EFFECT OF TEMPERATURE DURING YEAST FERMENTATION 1.1 Introduction Fermentation is defined as an energy yielding process where yeast converts sugar into energy, carbon dioxide or/and an alcohol depending on the respiration pathway. Yeast can respire in anaerobic ally and aerobically. However, yeast gets more energy from aerobic respiration, but in the absence of oxygen it can continue to respire anaerobic ally, though it does not get as much energy from the substrate. Yeast produces an alcohol when it respires anaerobic ally and ultimately the alcohol will kill the yeast. However the yeast fermentation process is affected by various process conditions and this determines the amount alcohol produced.

1.2 Purpose The purpose of this activity is to examine the influence of temperature on yeast fermentation

1.3 Objective Determine the influence of temperature on the yeast fermentation

1.4 Materials Ice water (0.0C) Bakers yeast Table sugar (sucrose) Thermometer Two 100 mL graduated cylinders Water (40C)

1.5 Procedure 1. Add 3.0 grams of table sugar to each 100 mL-graduated cylinder. 2. Add 2.0 grams of active dry yeast to each 100 mL-graduated cylinder.
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3. Add 50 mL of ice water to one graduated cylinder. Label the cylinder as ice temperature. 4. Add 50 mL of 40C water to the other graduated cylinder. Label the cylinder as 40C temperature. 5. Cap each cylinder with the palm of your hand and shake the cylinder vigorously. 6. Note the time. 7. Make observations for temperature change every five minutes for 20 minutes.

1.6 Questions 1. Calculate the amount of alcohol produced 2. Calculate the moles of CO2 produced 3. Explain if different sugars give equivalent number of CO2 moles 4. Formulate a conclusion based on your observations. 5. Outline other factors can affect the yeast fermentation process [3 marks] [3 marks] [3 marks] [5 marks] [11 marks]

2.0 DISSOLVED OXYGEN MEASUREMENT 2.1 Introduction The DO determination measures the amount of dissolved (or free) oxygen present in wastewater. Aerobic bacteria and aquatic life such as fish must have DO to survive. Aerobic wastewater treatment processes use aerobic bacteria to break down the organic compounds found in wastewater into more stable products that will not harm the receiving waters. Wastewater treatment facilities such as lagoons or ponds, trickling filters and activated sludge plants depend on these aerobic bacteria to treat sewage. The same type of aerobic wastewater treatment process occurs naturally in streams and ponds if organic matter is present, turning these bodies of water into aerobic wastewater treatment plants. If sufficient oxygen is not naturally supplied through wind and turbulence to replace the depleted oxygen, the body of water will develop a low DO and become anaerobic (or septic). The results of septic water bodies include fish kills and anaerobic odors. If the amount of free or DO present in the wastewater process becomes too low, the aerobic bacteria that normally treat the sewage will die. The process will not operate efficiently and septic conditions will occur. The DO test is used to monitor the process to

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ensure that there is enough dissolved oxygen present to keep the process from becoming septic. 2.2 Purpose The purpose of this practical is to find out the amount of dissolved oxygen in water.

2.3 Objective To determine the amount of dissolved oxygen in water. 2.4 Materials Manganous sulfate solution Alkaline potassium iodide-sodium azide solution Sulfuric acid (H2SO4), concentrated Starch indicator solution Sodium thiosulfate (Na2S2O3 5H2O), 0.025 N Phenylarsine oxide (PAO), 0.025 N Potassium bi-iodate (KH(IO3)2), 0.025 N Distilled or deionized water

These reagents are poisonous and should be handled with extreme caution. These reagents are corrosive and should be handled with extreme caution. 2.5 Equipment Burette, graduated to 0.1 mL Burette stand 300 mL glass stoppered BOD bottles 500 mL wide-mouthed Erlenmeyer flasks Pipettes with elongated tips and minimum volume of 1.0 mL (+/- 0.1 mL) Pipette bulb 250 mL graduated cylinders Distilled water rinse bottle

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2.6 Procedure 1. Collect the sample to be tested in a 300 mL BOD bottle taking special care to avoid adding air to the liquid being collected. Fill bottle completely and add stopper. Remove bottle stopper and add 1 mL of the manganous sulfate solution at the surface of the liquid. Add 1 mL of the alkaline-potassium iodide-sodium azide solution at the surface of the liquid. Replace the stopper, avoid trapping air bubbles and shake well by inverting the bottle several times. Repeat shaking after floc has settled halfway. Allow floc to settle a second time. Add 1 mL of concentrated sulfuric acid by allowing the acid to run down the neck of the bottle above the surface of the liquid. Restopper, rinse the top of the bottle to remove any acid and shake well until the precipitate has dissolved. Titrate a volume of treated sample which corresponds to 200 mL of the original sample. This corrects for the loss of some sample during the addition of reagents. This volume calculated using the formula: mL of sample to titrate = 200 x [300/(300-2)] = 201 mL Pour 201 mL of sample from the BOD bottle into an Erlenmeyer flask.

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NOTE: Since variations occur in the actual volume of each BOD bottle, do not pour 99 mL of sample out of the BOD bottle and assume that 201 mL will be left. If the solution is reddish-brown in colour, titrate with 0.0250N sodium thiosulfate or 0.0250 N PAO until the solution is a pale yellow (straw) colour. Record the amount of titrant used. Add a small quantity of starch indicator. If the solution has no reddish-brown colour, or is only slightly coloured, add a small quantity (approximately 1 mL) of starch indicator. If no blue colour develops, there is zero dissolved oxygen. If a blue colour develops. Titrate with 0.0250N sodium thiosulfate or 0.0250N PAO to the first disappearance of the blue colour. Record the total number of mL of sodium thiosulfate or PAO used.

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2.7 Questions 1. Calculate the amount of dissolved oxygen obtained from the sample [3 marks] 2. Calculate the concentration of the dissolved oxygen in the sample [3 marks] 3. Explain the reaction involved in the determination of DO in water using the Winkler method [3 marks]. 4. Explain how a change in the amount DO affect flora and fauna in the water body [2 marks] 5. Explain how warming or cooling of the atmosphere affect the amount of dissolved oxygen in your water [3 marks] 6. Explain how atmospheric pressure affect your DO readings [3 marks] 7. Explain how the amount of dissolved oxygen you measured agree with the amount you calculated. In addition, explain any deviations [3 marks].

REFERENCES Dunnivant FM, Hoboken, NJ Environmental Laboratory Exercises for Instrumental Analysis and Environmental, 2004. Common Operations and Wet Chemical Methods in Environmental Laboratories (this book). APHA-AWWA-WEF (1998) Effect of temperature on fermentation, evaluation copy of the vernier student lab, http://www.vernier.com/cmat/bwv.html.

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