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«Marine Potable Water Test Kit Instruction Booklet Page | 1 9th Sept. 2013 Marine Potable Water Test Kit Instruction Booklet Page 3. Safety and ...»

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Marine Potable Water Test Kit

Instruction Booklet

Page | 1 9th Sept. 2013

Marine Potable Water Test Kit

Instruction Booklet

Page

3. Safety and Hygiene

5. Notes page

6. Chlorine (free, combined and total)

7. pH

8. Chlorine (high range)

9. Coliforms and E.Coli bacteria

11. Total Bacteria Count

13. Iron

14. Copper

15. Colour

15. Turbidity

16. Enterococci bacteria 17 - 24. Portable Incubator User Instructions 25. Spares and consumables Page | 2 9th Sept. 2013 Safety and Hygiene Workplace and Cleanliness Simple to follow, step by step instructions are provided for the operation of all items and performance of all tests within this kit. It is however important to find a suitably clean and uncluttered area in which to organise and perform each test. Working in this environment will prevent mistakes and avoid contamination of the tests being performed. Ensure that all used items are suitably disinfected and disposed of correctly. Work surfaces should be cleaned with detergent and water before and after testing.

Personal Protective Equipment (PPE) Due to the nature of the test chemicals contained within this test kit and the potential hazard to health of the samples being tested, it is important to wear gloves and safety glasses when carrying out all tests contained within the kit.

Operational hazards Colitag and Bacteria Plates (unused) Unused Colitag (for the measurement of Coliforms/E.Coli) and Bacteria Plates present no hazard to health.

Colitag and Bacteria Plates (used) Bags or plates containing used/incubated samples for the analysis of E.Coli/Coliforms (thermotolerant coliforms) or TVC bacteria should be treated as potentially hazardous to health until they have been suitably disinfected (Disinfection Procedure below) and washed/rinsed. These samples could contain cultured bacteria and care should be taken NOT to allow the contents to come into contact with skin or eyes. The health risk is primarily from the ingestion of bacteria after transfer to clothing or hands, not from actual skin contact. If contact is made, remove contaminated clothing and wash thoroughly with a strong detergent. Wash contaminated skin with soap and water.

Samples Being Tested Again samples being tested could contain Coliform/E.Coli or TVC bacteria and, as such, present a similar risk to used Colitag or Bacteria Plate samples. As such, take the same precautions as detailed above.

Page | 3 9th Sept. 2013 Disinfection Procedure It is necessary to disinfect spent containers used in the determination of E.Coli/Coliforms & TVC’s prior to disposal. It is essential to wear appropriate protective equipment prior to performing the disinfection procedure. This includes gloves, safety glasses and appropriate overalls.

1) Fill a 5 litre bucket with at least 1 litre of strong bleach solution (5% activity).

2) Transfer all spent sampling bags, syringes and Bacteria Plates to the bucket.

3) Leave to sterilise for at least 2 hours.

(Ensure that the bleach solution is in contact will all sides of the containers, inside and out, leaving no air pockets).

4) Pour the bleach solution to waste and dispose of the equipment as any normal waste.

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Free chlorine (DPD No. 1 (R)-tablet)

1) Fill both cells to the 10 ml mark and

2) Place one cell in the left-hand compartment of the comparator as a blank.





3) Add one DPD No. 1 (R)-tablet in the other cell, close it with a lid and

4) Swirl it until the tablet has dissolved.

5) Place this second cell in the right-hand compartment of the comparator

6) Match the two colour fields against good light source and read off the result in mg/l free chlorine.

Total and combined chlorine (with the addition of DPD tablet No. 3 (R) Having completed the test for free chlorine,

7) Take the cell from the right-hand compartment of the comparator, remove the lid and add one DPD No. 3 (R) - tablet.

8) Replace the lid and swirl the cell until the tablet is dissolved.

9) Place the cell in the right-hand compartment and

10) Allow for 2 minutes reaction time. Then compare the two colour fields against a good light source and read off the result as total chlorine in mg/l.

Combined chlorine Deduct the reading for free chlorine from the figure obtained for total chlorine.

The difference represents the result for combined chlorine.

–  –  –

Important: Place the cell with the mark (point) to the viewer in the compartment. It is essential to rinse the cells thoroughly after each test. To obtain maximum accuracy always view and match colour against a good light source.

Store CHECKIT® Disc in the dark.

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Rapid dissolving PHENOL RED tablets (green printed foil = Type Rapid (R) pH (PHENOL RED (R)-tablet)

1) Fill both cells to the 10 ml mark.

2) Place one cell in the left-hand compartment of the comparator as a blank.

3) Add one pH (PHENOL RED (R))-tablet in the other cell, close it with a lid.

Swirl it until the tablet has dissolved.

4) Place this second cell in the right-hand compartment of the comparator.

5) Match the two colour fields against good light source and read off the result as pH.

Note:

pH values below 6.5 always produce a yellow colouration. pH values above 8.4 always produce a red colouration. Water samples with low values of Total Alkalinity may give wrong pH readings.

Important: Place the cell with the mark (point) to the viewer in the compartment. It is essential to rinse the cells thoroughly after each test. To obtain maximum accuracy always view and match colour against a good light source.

Store CHECKIT® Disc in the dark.

–  –  –

Chlorine High Range (CHLORINE HR / ACIDIFYING GP-tablet)

1) Fill both cells to the 10 ml mark.

2) Place one cell in the left-hand compartment of the comparator as a blank.

3) Add one CHLORINE HR-tablet to the other cell and crush to dissolve the tablet.

4) Add one ACIDIFYING GP-tablet to the same cell and crush to dissolve the tablet.

Close the cell with a lid. Swirl it until the tablet is dissolved.

5) Place this second cell in the right-hand compartment of the comparator.

6) Match the colour fields against good light source and read off the result as mg/l chlorine.

–  –  –

Coliforms and E.Coli Bacteria (Presence/Absence) - Order Code: 777932 This test procedure uses the Colitag sachets, 100ml sample bags and the handheld UV lamp. The test will take 24 hours to perform. It is important to store the samples at 35˚C during this period. This test is very sensitive and, as such, it is important to prevent contamination. Ensure that the person performing the sampling and testing has washed hands thoroughly with soap and water. Do not allow the bag to remain open for any period of time, other than to take the sample and add the Colitag sachet. Do not let the sample point touch the bag.

1) Take one of the 100ml sample bags and collect 100ml of water to be tested.

Use the pictorial guide for proper sampling procedure with the thio bags.

Note: It is important that the sample point is free from potential contamination. If necessary clean the sample point thoroughly prior to sampling.

The sample bag contains a tablet to neutralise any chlorine that may be present in the sample. Allow this tablet to dissolve before continuing to step 2.

2) Carefully add the contents of one Colitag Sachet to the 100ml sample.

Agitate gently to aid dissolution.

3) Place the sample inside one of the two plastic incubation pots provided and screw on the cap.

4) Incubate the sample at 35.0˚C +/-0.5˚C for 24 +/- 2 hours.

Note: Liquid samples MUST be incubated inside one of the incubation pots provided to prevent water entering the incubator should the liquid leak. Make sure the incubation pot is placed inside the incubator (with the lid off) during incubator warm-up period. Two samples can be placed inside one pot.

5) Sample interpretation a. Visually check the sample for yellow colour. If sample is yellow, then Coliform bacteria are present.

b. Place the sample in a dark area and expose to long wave UV light by shining the UV lamp supplied on the sample. If the sample fluoresces (glows a blue colour), E.Coli bacteria are present.

c. If no yellow colour in the test sample is observed after the 24 +/- 2 hour incubation period, the sample should be recorded as negative for Coliform bacteria and E.Coli (zero CFU/100ml) d. Product Storage: Store at 4˚C to 30˚C, (preferably 4˚C to 7˚C) away from the light. Observe the expiry dates on the packaging.

–  –  –

This test is used for the quantification of total aerobic bacteria in water.

The sample is added to dehydrated culture media and incubated for 48 hours at 35oC +/- 2oC (AOAC certified method). Aerobic bacteria show as red spots on the culture media and are counted to obtain the colony count as Colony Forming Units/ml (CFU/ml).

Storage Store bacteria plates between 1oC and 30oC and observe the expiry dates on the packaging. Store dilution pots between 1oC and 30oC and observe the expiry dates on the packaging.

Sample Preparation The collection bags supplied should be used to collect samples for total bacteria plate testing. The same sample can be used for this and the Coliform/E.Coli test.

The sample bag contains a sodium thiosulphate tablet to remove any chlorine present in the sample. However, a clean sterile collection vessel of any type may be used as the total bacteria plate contains a chlorine neutralising agent.

Test Range: 0- 2000 CFU/ml Procedure

1) Take one of the sterile,100ml sample bags and collect 100ml of water to be tested.

Use the pictorial guide for proper sampling procedure with the thio bags. The same sample can be used for the E.Coli/Coliform test.

Note: It is important that the sample point is free from potential contamination. If necessary clean the sample point thoroughly prior to sampling. The sample bag contains a tablet to neutralise any chlorine that may be present in the sample. Allow this tablet to dissolve before continuing to step 2.

2) Have the following items ready for use:

1 x 100ml sample bag containing the sample to be tested 2 x new, sealed 1ml syringes 1 x sterile alcohol wipe 1 x 9ml dilution pot 1 x TVC bacteria plate The incubator should be switched on and allowed to reach 35oC.

3) Wipe the foil surface of the dilution pot with the alcohol wipe and set the pot to one side (discard the wipe afterwards).

4) Using a new sterile 1ml syringe, draw up exactly 1ml of sample water from the sample bag.

–  –  –

5) Break the foil seal of the dilution pot with the base of the 1ml syringe and transfer the 1ml of sample into the pot. Remove the syringe and discard.

Swirl the pot gently to mix.

6) Take a new sterile 1ml syringe and fill with exactly 1ml of sample water from the dilution pot.

7) Open the lid of the TVC bacteria plate and slowly transfer the sample to the centre of the plate.

8) Cap the plate and transfer to the incubator.

9) Incubate at 35oC for 48 hours.

10) After 48 hours, place the plate on a white surface (or piece of paper) and count the number of red spot bacteria colonies on the surface of the plate.

11) The Colony Count (as CFU/ml) = Number of red spot colonies counted x10.

e.g. If 25 colonies are counted the final colony count for reporting purposes is 250 CFU/ml. Ideally, total bacteria levels in potable water should be less than 100 CFU/ml.

Guidance Notes for Plate Counting If the water sample is highly contaminated ( 200 growing colonies), the plate will not be able to show single colonies. The amount of chromogenic substrate in the media is not enough to colour every colony. The plate will normally turn a complete pale pink colour, not showing any individual colonies. In this case, report the result as 2000 CFU/ml.



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