1. Titlow Beach Marine Preserve An Introduction to Water Analysis Methods

2. Types of Tests Several types of tests that use scientific instruments Several types of tests that use scientific instruments Allow us to “see” chemicals in the water we can’t detect and other factors that affect the marine environment & its inhabitants Allow us to “see” chemicals in the water we can’t detect and other factors that affect the marine environment & its inhabitants

3. Direct Instrument Measurement Electrometric Method Electrometric Method pH Meter - pH pH Meter - pH Salinometer Salinometer Temperature Temperature Salinity Salinity Turbidimeter – Turbidity Turbidimeter – Turbidity

4. Titration Analysis Method Dissolved Oxygen (O 2 ) Dissolved Oxygen (O 2 ) Carbon Dioxide (CO 2 ) Carbon Dioxide (CO 2 ) Titrating solution added to treated sample until color change Titrating solution added to treated sample until color change Volume of titrant (chemical) proportional to concentration of chemical being test Volume of titrant (chemical) proportional to concentration of chemical being test

5. Colorimetric Analysis Nitrates (NO 3 - ) Nitrates (NO 3 - ) Phosphates (PO 4 -3 ) Phosphates (PO 4 -3 ) Sample treated with reagent Produces color reaction in proportion to amount of chemical

6. Titlow Beach Marine Preserve About 1 mile of shoreline About 1 mile of shoreline Managed by Park District of Tacoma Managed by Park District of Tacoma Sampling site = Park beach Sampling site = Park beach

7. Organizing Data Organization is key to any good study of science Organization is key to any good study of science Each monthly water collection entry in your lab books should have at least the following info: Each monthly water collection entry in your lab books should have at least the following info: Date & Time of sample collected Date & Time of sample collected Types of samples collected Types of samples collected Weather conditions & tides Weather conditions & tides Both individual & group average data Both individual & group average data

8. How to Label Lab Book Label the top of each NEW page with the following labels: Label the top of each NEW page with the following labels:

9. How to Label Lab Book (cont.) Water Sampling Collection Water Sampling Collection pH pH Temperature & Salinity Temperature & Salinity Dissolved Oxygen (D.O.) Dissolved Oxygen (D.O.) Carbon Dioxide (CO 2 ) Carbon Dioxide (CO 2 ) Nitrates (NO 3 - ) Nitrates (NO 3 - ) Plankton Plankton Turbidity Turbidity Phosphates ( PO 4 -3 ) Phosphates ( PO 4 -3 )

10. Organizing Data cont. Data should be organized so that others can understand & is easily used Data should be organized so that others can understand & is easily used Group Temp (°C) Salinity (‰) pH 1 2 3 4 5 6 7 Range Avg.

11. RULE OF THREE Rinse container and discard with sample water twice Rinse container and discard with sample water twice Keep the third sample Keep the third sample Water Sampling & Analysis

12. Water Sampling Collection

13. Water Sampling Collection cont. Plankton Net Plankton Net Plankton

14. pH

15. Salinity ( ‰) & Temperature (°C) YSI 85 Salinometer

16. Dissolved Oxygen Test Winkler Titration Method Two Parts Two Parts 1. Fixation Procedure (“Pickling”) 1. Fixation Procedure (“Pickling”) 2. Titration 2. Titration

17. 1. Add 2 mL of manganeese sulfate (MnSO 4 ) about 1 cm below the surface of the water. Mix and let precipitate settle. 1. Add 2 mL of manganeese sulfate (MnSO 4 ) about 1 cm below the surface of the water. Mix and let precipitate settle. 2. Add 2 mL of alkaline iodide (NaI) reagent about 1 cm below the surface of the water. Re-stopper BOD bottle and rock back and forth to mix thoroughly and let precipitate settle. 2. Add 2 mL of alkaline iodide (NaI) reagent about 1 cm below the surface of the water. Re-stopper BOD bottle and rock back and forth to mix thoroughly and let precipitate settle. Fixation Procedure (“Pickling”)

18. Fixation cont. 3. Add 2 mL of concentrated sulfuric acid (H 2 SO 4 ) allowing acid to run down the inside of the neck of bottle. Re- stopper and rock back and forth to mix. Precipitate will disappear and an orangish-brown solution will remain. 3. Add 2 mL of concentrated sulfuric acid (H 2 SO 4 ) allowing acid to run down the inside of the neck of bottle. Re- stopper and rock back and forth to mix. Precipitate will disappear and an orangish-brown solution will remain.

19. Dissolved Oxygen (O 2 ) “Pickling” Summary Add 2 mL of manganesse solution Add 2 mL of manganesse solution Add 2 mL of iodine solution Add 2 mL of iodine solution Add 2 mL of conc. Sulfuric acid Add 2 mL of conc. Sulfuric acid

20. Dissolved Oxygen (O 2 ) Titration setup Titration setup Buret with Phenylarsine Oxide (PAO) solution Buret with Phenylarsine Oxide (PAO) solution Add 201 ml of “pickled “ sample in 250 mL flask Add 201 ml of “pickled “ sample in 250 mL flask Add magnetic “bean” Add magnetic “bean”

21. Titration Procedure Record initial PAO level Record initial PAO level Turn on magnetic stirrer and slowly titrate sample with PAO solution. Turn on magnetic stirrer and slowly titrate sample with PAO solution. Titrate until solution turns a pale straw color. Titrate until solution turns a pale straw color. Add a dropper (2mL) of starch solution. Add a dropper (2mL) of starch solution.

22. O 2 Titration Procedure (cont.) Titrate drop by drop until the blue indicator first disappears (end point). Titrate drop by drop until the blue indicator first disappears (end point). Record the final level of PAO solution. Record the final level of PAO solution. The amount of dissolved oxygen is equal to the amount of PAO used and measured in mg/L or ppm. The amount of dissolved oxygen is equal to the amount of PAO used and measured in mg/L or ppm.

23. PRACTCIE TITRATION PRACTCIE TITRATION

24. Determining the % Oxygen Saturation Locate your temperature and salinity values on the nomogram Locate your temperature and salinity values on the nomogram Use a straight edge to connect the two values Use a straight edge to connect the two values Record the O 2 solubility to the nearest tenth Record the O 2 solubility to the nearest tenth Find the quotient of D.O./O 2 solubility Find the quotient of D.O./O 2 solubility Multiply by 100 to put into % Multiply by 100 to put into %

25. Carbon Dioxide (CO 2 )

26. Carbon Dioxide (CO 2 ) Methods 1. Fill titration bottle with 20 mL of water sample 1. Fill titration bottle with 20 mL of water sample 2. Add 2 drops of Phenolthalein. If sample turns red, no free CO 2 is present. If colorless, go to step 3. 2. Add 2 drops of Phenolthalein. If sample turns red, no free CO 2 is present. If colorless, go to step 3. 3. Fill Direct Reading Titrator with CO 2 Reagent. Insert Titrator in hole of titration tube cap. 3. Fill Direct Reading Titrator with CO 2 Reagent. Insert Titrator in hole of titration tube cap.

27. Carbon Dioxide Methods (cont) 4. While gently swirling, add reagent drop by drop until a faint pink color develops (must persist at least 30 seconds) 4. While gently swirling, add reagent drop by drop until a faint pink color develops (must persist at least 30 seconds) 5. Read where plunger tip meets Titrator scale. Record as ppm. 5. Read where plunger tip meets Titrator scale. Record as ppm.

28. Plankton

29. Plankton “Pickling” 1. Draw 3 mL of sample water into graduated test tube using pipette 1. Draw 3 mL of sample water into graduated test tube using pipette 2. Add 2 drops of iodine solution 2. Add 2 drops of iodine solution 3. Let stand overnight or centrifuge for 3:00 minutes. 3. Let stand overnight or centrifuge for 3:00 minutes.

30. Plankton Analysis 1. Pipette off the top 2 mL of pickled sample and discard 1. Pipette off the top 2 mL of pickled sample and discard 2. Place one drop of sample onto depression slide and examine 2. Place one drop of sample onto depression slide and examine 3. Examine drop by drop to completion 3. Examine drop by drop to completion

31. Plankton Analysis: The Microscope The MicroscopeThe Microscope

33. Microscope Care Always carry with 2 hands Always carry with 2 hands Only use lens paper for cleaning Only use lens paper for cleaning Do not force knobs Do not force knobs Always store covered Always store covered Keep objects clear of desk and cords Keep objects clear of desk and cords

34. Microscope Use Begin with stage up on low power objective Begin with stage up on low power objective Use course adjust for initial focus Use course adjust for initial focus Adjust diaphragm Adjust diaphragm Use fine adjust for final focus Use fine adjust for final focus Change to higher objective and focus with fine adjust Change to higher objective and focus with fine adjust

35. Plankton Labeled drawings of each organism (determine type if possible) Labeled drawings of each organism (determine type if possible) Total magnification Total magnification Diaphragm setting Diaphragm setting Plankton Shape Plankton Shape Occurrence (count # of times seen) Occurrence (count # of times seen)

36. Plankton Shapes Circles Circles Rectangles/boxes Rectangles/boxes Spirals Spirals Chains (circle or box) Chains (circle or box) Spheres (maybe it’s spears!) Spheres (maybe it’s spears!) Animal Animal Other Other

37. Nitrate (NO 3 - ) Test Methods 1. Fill tube to 2.5 mL mark with sample. 1. Fill tube to 2.5 mL mark with sample. 2. Add 2.5 mL of Mixed Acid Reagent, cap and mix. Wait two minutes. 2. Add 2.5 mL of Mixed Acid Reagent, cap and mix. Wait two minutes. 3. Add 0.1 grams (1 level spoonful) of Nitrate Reducing Reagent. Cap and invert 50-60 times in a minute. Wait ten minutes. 3. Add 0.1 grams (1 level spoonful) of Nitrate Reducing Reagent. Cap and invert 50-60 times in a minute. Wait ten minutes.

38. Nitrate Test Methods (cont.) 4. Use Octa-Slide Viewer and match sample color to color standard. 4. Use Octa-Slide Viewer and match sample color to color standard.

39. Turbidity Rinse the inside of a clean cuvette three times with the sample to be tested keeping the third rinse. Rinse the inside of a clean cuvette three times with the sample to be tested keeping the third rinse. Clean outside of cuvette with Kimex. Clean outside of cuvette with Kimex. Insert cuvette into the opitical well and index to the lowest reading. Insert cuvette into the opitical well and index to the lowest reading. Record the NTU value. Record the NTU value.

40. Turbidity

41. Phosphates ( PO 4 -3 ) – Solution Prep Fill sample cell with 10 mL of sample Fill sample cell with 10 mL of sample Add the contents of one PhosVer 3 phosphate Powder Pillow to the cell. Stopper and shake vigorously for 30 seconds. Add the contents of one PhosVer 3 phosphate Powder Pillow to the cell. Stopper and shake vigorously for 30 seconds. Start the instrument timer and let sample react for 2 minutes Start the instrument timer and let sample react for 2 minutes Meanwhile, prepare the blank by filling a 2 nd sample cell with 10 mL of sample. Meanwhile, prepare the blank by filling a 2 nd sample cell with 10 mL of sample.

42. Phosphate Test When the timer expires, wipe the blank with a Kimex and insert into the cell holder When the timer expires, wipe the blank with a Kimex and insert into the cell holder ZERO the instrument (0.00 mg/L PO 4 3- will show) ZERO the instrument (0.00 mg/L PO 4 3- will show) Wipe the prepared sample and insert Wipe the prepared sample and insert Read the results in mg/L PO 4 3- (or ppm) Read the results in mg/L PO 4 3- (or ppm)