WINKLER METHOD FOR DISSOLVED OXYGEN ANALYSIS

DARAMAN, NOLI CARLO P., DAYSA, RAECHELLE S., DELOS SANTOS, JONALYN D., FACUN, ANGELI ELEANOR B. Department of Biology, College of Science, University of the Philippines Baguio August 3, 2012 ABSTRACT The main objective of this experiment is for the students to understand and apply the Winkler method of measuring the dissolved oxygen. This experiment also aims to undertake an iodometric or indirect method of analysis. Three trials of standardization of 0.05 N sodium thiosulfate and six trials of Winkler method were the main procedures done in the experiment. Overall, it can be concluded that this experiment was successful and all the objectives were satisfied.

INTRODUCTION Water plays a very significant role in the lives of every living thing on earth. Its value cannot be overlooked since it has been an important ingredient for the evolution of Earth. Water has been a source of oxygen for millions of living species. Oxygen is a very essential element since it aids in the aerobic respiration of all organisms, with a few exceptions, from the smallest bacteria up to the largest existing blue whales and giant red wood. The amount of dissolved oxygen present in water determines the diversity of organisms in a particular ecosystem especially those that are aquatic. It also determines water quality. Dissolved oxygen analysis measures the amount of gaseous oxygen (O2) dissolved in an aqueous solution. Oxygen gets into water by diffusion from the surrounding air, by aeration (rapid movement), and as a waste product of photosynthesis. Total dissolved gas concentrations in water should not exceed 110 percent. Concentrations above this level can be harmful to aquatic life. Fish in waters containing

excessive dissolved gases may suffer from "gas bubble disease"; however, this is a very rare occurrence. The bubbles or emboli block the flow of blood through blood vessels causing death. External bubbles (emphysema) can also occur and be seen on fins, on skin and on other tissue. (KY Water Watch, N.D.) This experiment utilizes the Winkler method for dissolved oxygen analysis. The Winkler Method is a technique used to measure dissolved oxygen in freshwater systems. Dissolved oxygen is used as an indicator of the health of a water body, where higher dissolved oxygen concentrations are correlated with high productivity and little pollution. This test is performed on-site, as delays between sample collections and testing may result in an alteration in oxygen content. (Bruckner, N.D.) The following reactions take place during the Winkler Method: 1. Manganese (II) ions liberated from the manganese sulfate are loosely bound with excess hydroxide. 2+ Mn + 2OH---> Mn(OH)2 2. Manganese (II) is oxidized to Manganese

(III) in the presence of a strong base and binds the dissolved oxygen. 2Mn(OH)2 + O2 +H2O ---> 2Mn(OH)3 3. Free iodine is produced upon acidification of the sample at a rate of one I2 molecule for each atom of oxygen. 2Mn(OH)3 + 2I- + 6H+ ---> 2Mn2+ + I2 + 6H2O 4. Free iodine complexes with excess iodide ions. I2 + I- ---> I35. The iodine/iodide complex is reduced to iodide with thiosulfate. I3- + 2S2O32- ---> 3I + S4O6 2- (Grasshoff et al., 1983) RESULTS Table 1.0 Standardization of KMnO4 Solution Trial 1 2 3 Weight of 0.1194 0.1087 0.1382 K2Cr2O7 used (g) Weight of 0.2501 0.2536 0.2508 Na2CO3 used (g) Weight of KI used 1.0155 1.0049 1.0024 (g) Thiosulfate 44.20 46.90 61.40 volume used (ml) Na2S2O3 molarity 0.05510 0.04727 0.04591 Na2S2O3 average molarity 0.04943

Dissolved Oxygen (Reference): 11.5 mg/L, 11.9 mg/L, 12.0 mg/L Average Dissolved Oxygen (Reference): 11.8 mg/L Table 3.0 Volume of Thiosulfate Used (mL) Trial 1 2 3 Bottle 1 (g/ml) 0.20 ml 0.20 ml 0.25 ml Bottle 2 (g/ml) 0.15 ml 0.15 ml 0.20 ml *Bottle 1 is 300 ml and Bottle 2 is 290 ml Table 4.0 Dissolved Oxygen Concentration Trial 1 2 3 Bottle 1 (g/ml) 15.81 15.81 19.76 Bottle 2 (g/ml) 11.86 11.86 15.81 Average Dissolved Oxygen: 15.15 mg/L DISCUSSION For the first part of the experiment which is the standardization of the thiosulfate, the balanced equation K2Cr2O7 + 6NaS2O3 + 14H+ ---> 2Cr3+ + 2K+ + 12Na+ + 3S4O6 + 7H2O was used for the calculation of the molarity of the thiosulfate. Potassium dichromate was used to standardize the sodium thiosulfate solution because it is a useful primary standard for redox reactions. It can be obtained in high purity, it has a high equivalent weight, it is nonhygroscopic, and the solid and its solutions are quite stable. Sodium carbonate was added to the solution which serves as the preservative. Starch was also added to determine precisely the endpoint and to determine the endpoint more easily. The mass of K2Cr2O7 that were used are 0.1194 g for trial 1, 0.1087 for trial 2 and 0.1382 for trial 3. The volume of the thiosulfate that was used are 44.20 mL, 46.90 mL and 61.40 mL, respectively. With these results, the molarity of

Collection and Analysis of Water Samples Table 2.0 Parameters Observed in the Field Weather condition: Cloudy Water condition Color: Green Mobility: Stagnant Odor: Unpleasant Turbidity: Unclear Location: Shady Temperature and pH of water Temperature: 22.21 C, 23.01C, 22.87C pH: 9.39, 9.40, 9.36 Time of the day: 3:17 pm

the Na2S2O3 for each trial was computed which 0.05510 M, 0.04727 M and 0.04591 M are averaging to 0.4943 M. On the second part of the experiment which is the analysis of the collected water which was analyzed with the use of the Winkler method, two bottles were used each having 3 trials were done. The dissolved oxygen that were computed were 18.51 mg/L for the first trial, 18.51 mg/L for the second trial, 19.76 mg/L for the third trial, 11.86 mg/L for the fourth trial, 11.86 mg/L for the fifth trial and 15.81 mg/L for the sixth trial. The computed average DO for the six trials is 15.15 mg/L. It can be noticed that the computed value for the third trial is far from the values of the other five values but it has been computed that the third trial is not an outlier. The computed average of the DO in the six trials which is 15.15 mg/L when compared to the average DO reference which is 11.8 mg/L do not really have the a significant difference which is due to some factors or parameters. The parameters that were observed in the field where the sample water was collected are cloudy for the weather condition; green, unpleasant, stagnant and unclear for the water condition; shady for the location, 22.21 degrees Celcius, 23.01 degrees Celcius, 22.87 degrees Celcius for the temperature and 9.39, 9.40, 9.36 for the pH of water; and 3:17 pm for the time of the day. These parameters affect the DO in some way which explains why the computed DO is greater than that of the reference DO. Example is for the temperature, wherein the lower the temperature, the higher the amount of dissolved oxygen in a body of water and vice versa. CONCLUSION

The experiment had shown that the dissolved oxygen is 15.15 mg/L was relatively high to the standards which are being prepared. Since the computed DO and the reference DO is higher than 10 mg/L (Scioto SWCD, N.D.) it can be concluded that the water found on the Burnham lake is still healthy which explains why a fish was seen during the collection of water samples.

LITERATURE CITED Bruckner, Monica Z. N.D. http://serc.carleton.edu/microbelife/resear ch_methods/environ_sampling/oxygen.htm l, accessed on 31 July 2012. Grasshoff, K. et al., 1983. Methods of Seawater Analysis: Second, revised and extended edition. New York. Verlag Chemie. KY Water Watch. N.D. http://www.state.ky.us/nrepc/water/wcpd o.htm, accessed on 31 July 2012. Scioto SWCD. N.D. http://www.sciotoswcd.org/testinfo.html, accessed on 31 July 2012.

APPENDIX A. Molarity of of Sodium Thiosulfate

Trial 1:

Trial 2:

Trial 3:

Trial 6:

Average molarity: Average DO:

B. Dissolved Oxygen Mean of the DO:

Trial 1: DO levels Deviation Trial 1 |15.15 15.81| 0.66 Trial 2 |15.15 15.81| 0.66 Trial 3 |15.15 19.76| 4.61 Trial 4 |15.15 11.86| 3.29 Trial 5 |15.15 11.86| 3.29 Trial 6 |15.15 15.81| 0.66 Average of D2 = 13.17/6 = 2.20 Variance = Trial 3: S.D. = POST-LABORATORY REQUIREMENTS Trial 4: 1. Use at least two statistical tools to evaluate the precision of your analysis. Evaluation of precision using variance: Trial 5: Mean of the DO: D2 0.4356 0.4356 21.2521 10.8241 10.8241 0.4356

Trial 2:

DO levels Deviation Trial 1 |15.15 15.81| 0.66 Trial 2 |15.15 15.81| 0.66 Trial 3 |15.15 19.76| 4.61 Trial 4 |15.15 11.86| 3.29 Trial 5 |15.15 11.86| 3.29 Trial 6 |15.15 15.81| 0.66 2 Average of D = 13.17/6 = 2.20 Variance = Evaluation deviation: S.D. = of precision using

D2 0.4356 0.4356 21.2521 10.8241 10.8241 0.4356

standard

parameter is Turbidity which is the condition resulting from suspended solids in the water, including silts, clays, industrial wastes, sewage and plankton. Such particles absorb heat in the sunlight, thus raising water temperature, which in turn lowers dissolved oxygen levels. They also prevent sunlight from reaching plants below the surface. This decreases the rate of photosynthesis, so less oxygen is produced by plants. Third is the Dissolved Oxygen level. It is said that the higher the dissolved oxygen, the healthier and cleaner the body of water.

2. Compare the DO levels obtained from all field sites. Make a generalization regarding the degree of pollution along the stream profile of the body of water that you worked on. The reference dissolved oxygen and the computed dissolved oxygen of the group in a part of the lake or field site has a high DO level as explained in the conclusion. Other field sites which where tested also show that it has a high DO level. The DO level from the different field sites ranges from 11 mg/L 15 mg/L. 3. Report on the degree of pollution of the body of water using at least three parameters including DO level. The following parameters affect the degree of pollution of the body of water. In temperature, the lower the temperature, the higher the amount of dissolved oxygen in a body of water; the opposite is also true, in other words, the higher the water temperature the lower the amount of dissolved oxygen. The amount of DO is highly dependent on the temperature. Next