LB (Luria Bertani):TryptoneYeast ExtractNaCLLB made up

Isosensitist MediaIsosensitist made up

MOPS20mM MOPS pH 7.0Hazardous Substances Policy - Assessment

CHEMICAL HAZARD AND RISK ASSESSMENT

School of Biosciences

Name of supervisor / Prof N Brown / Assessment Number* / 1987
Signature
Assessor / Rita Godfrey / Date of Assessment / 20/6/02

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 / T101
2 / PERSONS WHO MAY BE AT RISK
List names where possible / Post Docs, Post Graduates and Technicians in T101
3 / ACTIVITY ASSESSED / Protein Isolation Protocol 2
4 / MATERIALS INVOLVED / Attach copies of data sheet(s)
NAME / AMOUNT
max/stored / HAZARD / RISK PHRASES / HAZDAT NO*** / BIOSCIENCESNO***
LB (Luria Bertani):
Tryptone
Yeast Extract
NaCL
LB made up / 100g
100g
1kg / Irritant
Irritant
NSH
NSH / R37/38 Irritating to the respiratory system and skin
R37/38 Irritating to the respiratory system and skin / 1303
Isosensitist Media
Isosensitist made up / 500g / Irritant
NSH / R37 Irritating to the respiratory system
MOPS
20mM MOPS pH 7.0 / 50g / Irritant
NSH / R36/37/38 Irritating to eyes, respiratory system and skin
Tris
40mM Tris-HCl pH 9.5 / 1kg / Irritant
Harmful
NSH / R36/37/38 Irritating to the eyes, respiratory system and skin / 911
DNase / 0.1ml / Hazard unknown / Avoid contact and inhalation
RNase / 0.1ml / Hazard unknown / Avoid contact and inhalation
Solubilization Solution 2:
Urea
CHAPS
Tris
DTT
IPG buffer
DNAse
RNAse
Solubilization solution 2 / 500g
500g
1kg
25g
1ml
0.1ml
0.1ml
100ml / NSH
Irritant
See above
Irritant
NSH
See above
See above
NSH / R36/37/38 Irritating to the eyes, respiratory system and skin
See above
R36/38 Irritating to the eyes and respiratory system
See above
See above / 906
1272
1340
Solubilization Solution 3:
Urea
Thiourea
Chaps
SB3-10
Tris
TBP(tributyl phosphine)
IPG buffer
DNase
RNase
Solubilzation solution 3 / 500g
500g
500g
100g
1kg
20ml
1ml
0.1ml
0.1ml
20ml / See Above
Harmful
Category 3
Carcinogen
Dangerous for the Environment
See Above
Irritant
See Above
Corrosive
Highly Flammable
Harmful
NSH
See Above
See Above
Category 3
Carcinogen
Dangerous for the Environment / See Above
R22 Harmful if swallowed
R40 Possible risk of irreversible effects. R63 Possible risk of harm to the unborn child
R50/53 Very toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment
See above
R36/37/38 Irritating to the eyes, respiratory system and skin.
See Above
R34 Causes burns
R17 Spontaneously flammable in air
R21/22 Harmful in contact with skin and if swallowed.
See Above
See Above
R40 Possible risk of irreversible effects
R63 Possible risk of harm to the unborn child
R51/53 Toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment. / 867
Solubilization Solution 4:
SDS
Glycerol
Tris
DTT
Solubilization solution 4
Rehydration Solution A:
Urea
Chaps
Tris
TBP
IPG
Rehydration solution A
Rehydration Solution B:
Urea
Chaps
Tris
DTT
IPG
Rehydration solution B
Rehydration Solution C:
Urea
Thiourea
Chaps
SB3-10
Tris
TBP
IPG
Rehydration solution C
Rehydration Solution D:
contains rehydration solution C plus per 1.2ml
IPG buffer
TBP / 1kg
2.5litre
1kg
25g
10ml
500g
500g
1kg
20ml
1ml
10ml
500g
500g
1kg
25g
1ml
10ml
500g
500g
500g
100g
1kg
20ml
1ml
20ml
20ml
6ul
o.6ul / Harmful
Irritant
See Above
See Above
Irritant
See Above
See Above
See Above
See Above
NSH
NSH
See Above
See Above
See Above
See Above
NSH
NSH
See Above
See Above
See Above
See Above
See Above
See Above
NSH
Category 3 Carcinogen
Dangerous for the Environment
Category 3 Carcinogen
Dangerous for the Environment
See Above
See Above / R22 Harmful if swallowed
R36/38 Irritating to eyes and skin
R36 Irritating to the eyes
See Above
See Above
R36 Irritating to the eyes
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
See Above
R40 Possible risk of irreversible effects
R63 Possible risk of harm to the unborn child
R51/53 Toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment.
R40 Possible risk of irreversible effects
R63 Possible risk of harm to the unborn child
R51/53 Toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment.
See Above
See Above / 966
354
5 / INTENDED USE**
Give brief details and attach protocol/instructions
Isolation of Membrane Proteins from E.Coli following attached protocol.
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?
TBP is extremely dangerous if exposed to oxygen.
There is a risk to the user from Thiourea, both from the solid form and to a lesser extent from the solutions that contain it through spillage. Pregnant women are especially at risk
There is a risk of inhaling dust from tryptone, yeast extract, isosensitist media, Tris, Chaps, DTT, and SB3-10
7 / CONCLUSIONS ABOUT RISKS
Is level of risk acceptable? Can risk be prevented or reduced by change of substance/procedure? Are control measures necessary?
The risk from the stock solution of TBP is considerable and control measures are necessary. The risk from the solutions containing TBP is negligible as the concentration and amounts used are so low.
As the risk from the DNase and RNase is unknown it is wise to handle them with caution.
Certain control measures are also required when using the other chemicals that classified as hazardous.
The use and disposal of thiourea needs to be controlled. Pregnant women may choose not to work with this
8 / CONTROL MEASURES
Additional to Good Chemical Practice
Aliquoting of TBP needs to be carried out in a nitrogen chamber – a chamber that is flooded with nitrogen gas to eliminate oxygen. Once the TBP is in the solution it is safe. Only very small volumes are used (0.5ul) It is added to the solution after the solution has been sonicated. User of the sonicator must wear ear protectors and alert other members of the lab that it is being used.
The following compounds should be weighed out in a fume hood: Tryptone, yeast extract, isosensitist media, thiourea and SDS. Tris, CHAPS,DTT and SB3-10 can be weighed out on the open bench as long as care is taken not to generate dust.
Gloves should be worn when handling any of the chemicals mentioned in this section as well as with DNase and RNase.
9 / INSTRUCTION/TRAINING
Specify course(s) and/or special arrangements.
Instruction in the use of the Nitrogen gas chamber and the sonicator must be carried out before being used for the first time
10 / MONITORING
Performance of control measures,
Check fume hood is working before using to weigh out chemicals.
Check the nitrogen chamber is functioning properly
Check integrity of gloves
Personal exposure / Health Surveillance
Occupational Health must be notified before commencing this work due to the presence of a carcinogen. Tel:42695
11 / WASTE DISPOSAL PROCEDURE
See School Server for Approved Procedure Document on specific Chemical Waste Disposal.
Thiourea and all solutions containing it MUST NOT go down the sink. It should be stored in the non-halogenated waste solvent bottle until full and then taken down to Stores for specialist disposal.
Solutions containing TBP can be washed down the sink with plenty of water as the concentration of TBP and the volumes used are small. Any waste concentrated TBP would have to be disposed of as specialist waste. Bryan Facer in the Stores should be contacted regarding its disposal.
All the other compounds can be washed down the sink with copious amounts of water.
12 / REVIEW
Enter the date or circumstances for review of assessment (maximum review interval 5 years)
Every 12 months
13 / EMERGENCY ACTION
TO CONTROL HAZARDS / To stabilize situation eg spread absorbant on liquid spill; eliminate sources of ignition, etc.
Thiourea – evacuate the area. Wearing gloves, protective clothing and face mask sweep up and collect into a suitable labelled container. Take to Stores for specialist disposal.
TBP – evacuate area. Wearing gloves, protective clothing and face mask cover with absorbant powder from spill kit. Collect into a suitable leakproof, labelled container and take to Stores for specialist disposal.
All other chemicals used can be carefully swept or mopped up and placed in a suitable container for disposal.
TO PROTECT PERSONNEL / Evacuation, protection for personnel involved in clean-up, Special First Aid
Evacuation may be necessary for spillages of thiourea or TBP
Protective clothing, gloves ( rubber if the chemical is corrosive) and a face mask or goggles should be worn to clean up a spillage.
TO RENDER SITE OF EMERGENCY SAFE / Clean-up/decontamination
The site should be washed with water and detergent and mopped up with paper towels
CONTACT PHONE

10.10.00

* Prefix T is used for Teaching Assessment Number.

** Please include amount of chemicals used and how.

*** 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.