Hoechst 33258 dye

H33258 stock dye solution (1mg/ml)

10XTNE (100mM Tris, 10mM EDTA, 2M NaCl)Hazardous Substances Policy - Assessment

CHEMICAL HAZARD AND RISK ASSESSMENT

School of Biosciences

Name of supervisor / Dr S.D.Minchin / Assessment Number* / 1747
Signature
Assessor / Rita Godfrey / Date of Assessment / 10/11/00

Signature

Notes A School COSHH form in Word is available on the School Server.

Available from the Health and Safety Unit.

Guidance on making an assessment is given in Making a Chemical Hazard and Risk Assessment.

Guidance is also available from Guidance on Completing the Chemical Hazard and Risk Assessment Form.

Use a continuation sheet to expand any section of this form in hard copy version.

1 / LOCATION OF THE WORK ACTIVITY / Lab G10
2 / PERSONS WHO MAY BE AT RISK
List names where possible / All occupants of Lab G10 and users of the Hoefer DyNA Quant 200 Fluorometer
3 / ACTIVITY ASSESSED / Measurement of DNA concentrations using Hoechst 33258 dye in the Hoefer DyNA Quant flourometer.
4 / MATERIALS INVOLVED / Attach copies of data sheet(s)
NAME / AMOUNT** / HAZARD / RISK PHRASES / HAZDAT NO*** / BIOSCIENCESNO***
Hoechst 33258 dye / Irritant
Harmful / R36/37/38 Irritating to eyes, respiratory system and skin
R40 Possible risk of irreversible effects
H33258 stock dye solution (1mg/ml) / 50-100ml
contains
50-100mg / NSH
10XTNE (100mM Tris, 10mM EDTA, 2M NaCl) / 1 litre / NSH
Solids: Tris base / 12.11g / Irritant / R36/37/38 Irritating to eyes, respiratory system and skin / 911
EDTA / 3.72g / Harmful / R22 Harmful if swallowed / 1252
Sodium Chloride / 116.89g / NSH
Conc HCl
Calf thymus DNA, 1mg/ml
Assay solution A
Assay solution B / No more than 20ml
1mg
20ml
contains
2ug H33258
20ml
contains
20ug
H33258 / Corrosive
NSH
NSH
NSH / R34 Causes burns
R37 Irritating to respiratory system / 316
5 / INTENDED USE
Give brief details and attach protocol/instructions / Quantitation of DNA in a sample using Hoechst 33258, a DNA
Binding fluorescent dye in the Hoefer DyNA Quant 200 flourometer. See attached sheet for method.
6 / RISKS to HEALTH and SAFETY from INTENDED USE
From personal exposure or hazardous reactions. Refer to OELs, flash points, etc., as appropriate. Are pregnant women, breast-feeding mothers especially at risk?
There is a risk of skin contact and/or inhalation of the H33258 powder when making up the assay solution .
A There is a risk of splashing conc HCl onto the skin when preparing 10XTNE. There is also a risk of skin
Contact with Tris when weighing out. There is so little of the H33258 dye in the assay solutions to pose
Any great risk to health and safety. There is obviously more dye in the stock solution and the amount is still
Very low so there shouldn’t be a risk but due to the nature of the dye (DNA binding) care should be taken.
The amounts of Tris and EDTA in the 10XTNE are too small to have any risk.
7 / CONCLUSIONS ABOUT RISKS
Is level of risk acceptable? Can risk be prevented or reduced by change of substance/procedure? Are control measures necessary?
With proper control measures (outlined below) and good chemical practice the risks to health and safety are minimum
8 / CONTROL MEASURES
Additional to Good Chemical Practice
The H33258 dye should be weighed out in a fume hood whilst wearing protective clothing and gloves. .
Gloves should also be worn when handling solid Tris and when pH-ing the 10XTNE.
9 / INSTRUCTION/TRAINING
Specify course(s) and/or special arrangements.
None required
10 / MONITORING
Performance of control measures,
Check that the fume hood is working before weighing out the H33258. Check integrity of gloves before and during use.
Personal exposure / N/A / Health Surveillance / N/A
11 / WASTE DISPOSAL PROCEDURE
See School Server for Approved Procedure Document on specific Chemical Waste Disposal.
HCl – small quantities dilute to less than 5% in water. Wash down sink with plenty of water. Large quantities, add a little at a time, with stirring to an excess of water, to give a conc’n of less than 2%. Neutralise with sodium carbonate or 5% NaOH. Wash down sink with plenty of water.
Tris – Place in a seal-able container and dispose of as special waste. See School safety officer.
EDTA – Add a little at a time, with stirring to an excess of water. When reaction if any is complete, pour down sink with plenty of water.
Solid H33258 – as for Tris
H33258 stock solution – waste non-halogenated solvent bottle
Assay solutions – pour down sink with plenty of water.
12 / REVIEW
Enter the date or circumstances for review of assessment (maximum review interval 5 years)
November 2005
13 / EMERGENCY ACTION
TO CONTROL HAZARDS / To stabilize situation eg spread absorbant on liquid spill; eliminate sources of ignition, etc.
Fire extinguishers that may be used in an emergency are water spray, carbon dioxide, dry chemical powder or foam
TO PROTECT PERSONNEL / Evacuation, protection for personnel involved in clean-up, Special First Aid
For spillages of Assay solution, stock dye solution, solid H33258,Tris, EDTA and conc HCl wear protective clothing and gloves.
TO RENDER SITE OF EMERGENCY SAFE / Clean-up/decontamination
Spills of solid H33258 and Tris should be carefully swept up avoiding creating a dust and the powder
Transferred to a seal-able container for disposal after contacting the School Safety Officer. Thoroughly
Wash the contaminated area with lots of water and detergent. The site of an EDTA spillage should be
Washed with lots of water also. For HCl use inert spill absorbent, Na bicarbonate, Na carbonate,
Ca carbonate or lime to soak up the liquid spill. Ensure adequate ventilation as neutralising will produce CO2 gas.
CONTACT PHONE

* Prefix T is used for Teaching Assessment Number.

** List the amount by weight of the substance used. (for liquids eg; 100 mls 1M Sodium Hydroxide =

4g).

*** Hazdat No is the UNICOSHH datasheet report number.

Biosciences No is the Biosciences data sheet number.

UNICOSHH IS A CHEMICAL DATABASE ON THE HEALTH AND SAFETY UNIT SERVER.

BIOSCIENCES DATA SHEETS ARE AVAILABLE IN THE SCHOOL SAFETY OFFICE.