Eclox Manual

Section 4 Chemiluminescence Toxicity

Test

The Chemiluminescence Toxicity Test uses the luminometer. Before doing the

Chemiluminescence Toxicity Test, read

section 3.1, Overview on page 13

and do the procedures in

section 3.2, Prepare the luminometer for use on page 14

.

4.1 Overview

The Chemiluminescence Toxicity Test and Luminescent Bacteria Toxicity Test both show the inhibitory effects of a sample. However, the Chemiluminescence Toxicity

Test reagents are more rugged than the Luminescent Bacteria Toxicity Tests reagent and can be used under conditions where the Luminescent Bacteria Toxicity

Tests reagent can not be used.

The Chemiluminescence Toxicity Test reagents are stable for months even if stored

under higher ambient temperatures up to 40 °C ( Table 1

). The Luminescent

Bacteria Toxicity Tests reagent can not be stored under those conditions.

4.2 Prepare the reagents for luminometer calibration and sample testing

Prepare the chemiluminescence test (CT) Reagents 2 and 3 at the beginning of deployment.

The chemiluminescence test (CT) Reagents 2 and 3 are temperature sensitive and degrade at high temperatures. For long-term storage, store the reagents in their stable forms. On the first day of testing, prepare the reagents for routine use.

Diluted reagents are stable for 72 hours. The life of the reagents is longer if the reagents are kept cool (e.g., in a refrigerator) and in a dark place. Before use, let the reagents get to ambient temperature.

Table 1 Chemiluminescence test reagent stability

Reagent

Reagent 1

Reagent 2 (stable form)

Reagent 2 (diluted form)

Reagent 3 (stable form)

Reagent 3 (diluted form)

Refrigerated in a dark place

12 to 18 months

12 to 18 months

12 to 18 months

12 to 18 months

12 to 18 months

Raised Temperatures

(+40 °C)

1 year

6 months

72 hours

4 months

72 hours

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4.2.1 Prepare CT Reagent 2

1. Remove the CT

Reagent 2 buffer and CT

Reagent 2 caps.

Note: Do not open the bottles in heavy winds. The reagent in the CT Reagent 2 is small.

Note: Do not touch the reagent.

2. Carefully put all of the

CT Reagent 2 buffer in to the CT Reagent 2 bottle.

3. Put the caps back on the bottles. Shake the CT

Reagent 2 bottle for 30 seconds. Let dissolve for

10 minutes before use.

4.2.2 Prepare CT Reagent 3

1. Remove the CT

Reagent 3 concentrate and

CT Reagent 3 caps.

Note: Make sure that the batch number for CT Reagent

3 is the same as the batch number used for CT Reagent

2.

2. Push the end of the pipet in to a clean 100 µL yellow pipet tip and remove from the box.

3. Push in the operating button on the top of the pipet to the stop.

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4. Put the tip in the CT

Reagent 3 concentrate

1 cm below the surface.

Slowly release the operating button to pull in the concentrate.

5. Put the tip in to CT

Reagent 3 and dispense the liquid by gently pushing in the operating button.

Put the pipet in the storage case.

Put the tip in to the liquid and then remove from the liquid.

6. Remove the tip from the pipet and put in the waste bag.

7. Put the cap on CT

Reagent 3.

Turn over the CT Reagent

3 bottle several times to mix the solution.

Note: An ice chest can be used in the field to extend the life of the reagent.

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4.3 Calibrate the luminometer

Calibrate the luminometer before doing the Chemiluminescence Toxicity Test and after preparing the reagents.

The luminometer needs to be calibrated with every new batch of chemiluminescence reagents.

To calibrate the luminometer:

1.

Push ON (green button) to apply power to the luminometer.

The luminometer does built-in tests that make sure that the electronics and software are operating correctly.

2.

When the built-in tests are done, push PROCEED.

The Main Menu is shown.

3.

Select ECLOX and push ENTER.

The ECLOX Main Menu is shown.

4.

Push ENTER to select Measure.

The Measure Menu is shown.

5.

Select Measure Reference and push ENTER.

6.

Open the luminometer lid and make sure a sample is not in the cell. Then close the lid.

7.

Push PROCEED.

The test status is shown. The test may go a few minutes before it is done.

8.

When all the cell tests are done, push PROCEED.

9.

Open the lid of the Cuvette and 1000 µL Pipette Tip Set.

10. Put one cuvette in the black cuvette holder.

11. Place a blue pipette tip on the blue pipette.

12. Completely push in the operating button on the pipette to the stop and put the pipette tip in deionized water about 1 cm below the surface.

13. Release the operating button slowly to pull in the deionized water in to the pipette.

14. Touch the pipette tip against the side of the deionized water bottle to remove any drops from the outside of the tip.

15. Place the pipette tip in to the cuvette and dispense the liquid in to the cuvette by gently pushing in the operating button to the stop.

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16. Put the pipette tip in to the cuvette and remove the pipette from the cuvette to remove any drops from the outside of the tip.

17. Remove the lids from the CT Reagents 1, 2 and 3.

18. Put a yellow pipette tip on to the yellow pipette.

19. Put 100 µL of CT Reagent 1, 2 and 3 in to the cuvette using the pipette. Use a new pipette tip for each reagent.

20. Open the lid of the luminometer.

21. Lift the cuvette from the holder and gently tap the cuvette two times to mix the solution.

22. Immediately put the cuvette in to the luminometer cell and close the lid.

23. Push PROCEED.

The luminometer automatically starts measuring. After four minutes, the screen count down timer displays DONE.

24. When the measurement is complete, remove the cuvette from the luminometer.

25. Put the solution in the cuvette in to the waste bottle.

26. Put the cuvette in to the waste bag.

27. If the reference is between 300 and 900, the calibration is complete.

28. If the reference is not between 300 and 900, push PROCEED and do the calibration procedure again.

Note: New reagents may give a signal over 900. If the signal is 900 or over, change the measurement range to the 0–2000 range. Do not throw away the reagent set. If the signal is below 300, the reagents are probably unusable due to temperature sensitivity and new ones are required.

Note: If the reagent baseline is reading over 1000 or the luminometer shows Detector

Overload, change the measurement range to the 0–2000 range and continue. There is no need to throw away the chemiluminescent reagents.

29. If the signal is below 300 again, add another 100 µL of CT Reagent 3 to the cuvette and do the calibration procedure again.

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4.4 Measure pollutants in the water sample

Start with fresh reference everyday and with each new reagent set.

If the measured light units for the reagents are over 900, change the measurement range to the 0–2000 range and continue (refer to

section 3.2.3, Set the measurement range on page 16

).

1. Fill the beaker with

50 mL of sample water.

2. If more than 0.4 mg/L chlorine is present, neutralize the sample by adding two drops of pre-conditioner reagent to the sample beaker.

Note: Two drops of pre-conditioner reagent can neutralize up to 15 mg/L of chlorine.

3. Push

ON

(green button) for several seconds to apply power to the luminometer.

When the built-in tests are done, push PROCEED.

The Main Menu is shown.

4. Select ECLOX and push

ENTER.

5. Open the luminometer lid and make sure that a sample is not in the cell.

Close the lid.

Select Measure and push

ENTER.

Select Measure Sample and push ENTER.

6. Push PROCEED to show the test status.

When the cell tests are done, push PROCEED again.

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7. Put one cuvette from the

Cuvettes and 1000 µL

Pipet Tip Set in to the black cuvette holder.

8. Put a blue pipette tip on the blue pipet.

9. Push in the operating button on the pipet to the stop.

10. Put the tip in the sample water 1 cm below the surface. Slowly release the operating button to pull in the sample.

11. Put the tip in to the cuvette and dispense the liquid by gently pushing in the operating button.

12. Remove the tip from the pipet and put in the waste bag. Put the pipet in the storage case.

13. Put a yellow pipet tip on the yellow pipet.

Use a new pipet tip for each reagent.

14. Do steps 9 to 12 to put

100 µL of CT Reagents 1, 2 and 3 in to the cuvette.

15. Open the luminometer lid. Remove the cuvette from the cuvette holder.

Gently tap the cuvette twice to mix the solution. Put the cuvette in the luminometer cell.

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16. Close the lid. Push

PROCEED.

The luminometer automatically starts measuring. After four minutes, the screen timer shows DONE.

17. The Inhib% is shown on the screen. Record the

Inhib% value and graph on the Test Record Sheet.

Note: For sea water samples, the graph may appear higher than the reference and the

Inhib% may be negative.

18. Push PROCEED to go back to the Measure Menu.

19. Remove the cuvette from the luminometer cell.

Put the solution in to the waste bottle. Put the cuvette in to the waste bag.

20. Sign the Test Record

Sheet.

Put the sample from the beaker in the wastewater drain using local operating procedures.

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4.5 Show the previous results

Up to 60 previous results (samples plus references) and graphs can be saved on the luminometer and then shown later on the luminometer.

To save previous results to a computer, refer to

Send previous results to a computer on page 31

.

To show previous results saved on the luminometer:

1.

Push ON (green button) for several seconds to apply power to the luminometer.

The luminometer does built-in tests that make sure that the electronics and software are operating correctly.

2.

When the built-in tests are done, push PROCEED.

The Main Menu is shown.

3.

Select ECLOX and push ENTER.

4.

Select Previous Results and push ENTER.

The Previous Results Menu is shown.

5.

To show previous results:

a.

Select Recall Results and push ENTER.

b. Push MORE to show more results.

c.

Push QUIT to go back to the Previous Results Menu.

6.

To show previous graphs:

a.

Select Recall Graphs and push ENTER.

b. Push UP to move through the saved samples.

c.

Push SELECT when the required graph number is shown to select a graph.

d. Push SELECT again on the last selected graph number to select multiple graphs.

4.6 Send previous results to a computer

To send previous results to a computer:

1.

Do the steps in

Connect the luminometer to a computer on page 21

.

2.

Start LUMISsoft.

3.

In LUMISsoft, select Transfer, Options, Interface Protocol, Connect.

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4.

Push ON (green button) for several seconds to apply power to the luminometer.

The luminometer does built-in tests that make sure that the electronics and software are operating correctly.

5.

When the built-in tests are done, push PROCEED.

The Main Menu is shown.

6.

Select ECLOX and push ENTER.

7.

Select Previous Results and push ENTER.

The Previous Results Menu is shown.

8.

Select Download to PC and push ENTER.

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