Glycerol
10% Glycerol
Hazardous Substances Policy - Assessment
CHEMICAL HAZARD AND RISK ASSESSMENT
School of Biosciences
Name of supervisor / Dr S. Minchin / Assessment Number* / 1817Signature
Assessor / Rita Godfrey / Date of Assessment / 29 / 5 / 01
Signature
NotesA 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 G102 / PERSONS WHO MAY BE AT RISK
List names where possible / All occupants of the lab (Laura Tate, Jo Field, Dong Ling Zeng, Rita Godfrey)
3 / ACTIVITY ASSESSED / Preparing electrocompetent bacterial cells and their use in electroporations.
4 / MATERIALS INVOLVED / Attach copies of data sheet(s)
NAME / AMOUNT
max/stored / HAZARD / RISK PHRASES / HAZDAT NO*** / BIOSCIENCESNO***
Glycerol / 2.5 litres / IRRITANT / R36 Irritating to the eyes / 354
10% Glycerol / 800ml / NSH
Bacterial strain (E. coli) / 1colony / Hazard Group 1 under GMAG rules
Bacterial culture grown overnight / 5ml / Hazard Group 1 under
GMAG rules
Exponential phase bacterial culture / 400ml / Hazard Group 1 under GMAG rules
Plasmid DNA / mg-g qnty / Hazard Group 1 under GMAG rules
SOC media i.e
2% Bactotryptone
0.5% Yeast extract
10mM NaCl
2.5mM KCl
10mM MgCl2
10mM MgSO4
20mM Glucose / 200ml / NSH
100mg/ml Ampicillin / 5ml / HARMFUL / R42/43 May cause sensitization by inhalation and skin contact
LB amp plates i.e
1% Bactotryptone
0.5% yeast extract
0.5% NaCl
1.2% Agar
100ug/ml Ampicillin or other antibiotic / 1-2 litres worth are stored / NSH / None but avoid making dusts which may irritate the mucous membranes and the respiratory system
5 / INTENDED USE**
Give brief details and attach protocol/instructions / A culture of E.coli is grown up and then spun down resuspending
the pellet in a gradually smaller volume of 10% glycerol. See attached sheet. A small amount of cells are
then placed in a special electroporator cuvette along with the plasmid DNA of interest and a large voltage is
passed through them allowing the cells to take up the DNA. After incubating for a short while the cells are
plated onto selective media.
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 may be a risk of irritation of the mucous membranes and respiratory system from the fine powder
used to make SOC and the LB plates. There may be an electrocution risk from the electroporator machine.
There is a risk of irritation of the eyes from glycerol.
7 / CONCLUSIONS ABOUT RISKS
Is level of risk acceptable? Can risk be prevented or reduced by change of substance/procedure? Are control measures necessary?
Good Chemical Practice should cover the risks, although rough handling when weighing out the LB agar
components could produce a health risk. When unloading the autoclave there may a risk of pressurised
containers exploding or forcing the tops off. The electrocution risk should be considered.
8 / CONTROL MEASURES
Additional to Good Chemical Practice
When weighing out the media care should be taken so that a dust is not generated. Gentle handling should
avoid this. Make sure the tops of bottles of media are loose when they go into the autoclave and do not
open until the pressure has reached zero. Check the connections on the electroporator before beginning and
make sure that it is electrically tested regularly. Avoid touching the eyes when handling glycerol.
9 / INSTRUCTION/TRAINING
Specify course(s) and/or special arrangements.
10 / MONITORING
Performance of control measures,
Personal exposure / Health Surveillance
11 / WASTE DISPOSAL PROCEDURE
See School Server for Approved Procedure Document on specific Chemical Waste Disposal.
The supernatants produced after spinning the cells down should be treated with Presept disinfectant before
being poured down the sink. Waste agar plates that had E. coli grown on them have to be autoclaved
before they can be disposed of.
12 / REVIEW
Enter the date or circumstances for review of assessment (maximum review interval 5 years)
May 2006
13 / EMERGENCY ACTION
TO CONTROL HAZARDS / To stabilize situation eg spread absorbant on liquid spill; eliminate sources of ignition, etc.
Spills of glycerol should be mopped up with paper towels. Spills of bacterial cultures should be treated by
covering with Presept granules. Small spills can be treated by applying 70% ethanol to the area although
care must be taken to shut off all sources of ignition.
TO PROTECT PERSONNEL / Evacuation, protection for personnel involved in clean-up, Special First Aid
Protective clothing and gloves should be worn during the clean up procedure.
TO RENDER SITE OF EMERGENCY SAFE / Clean-up/decontamination
After the site of a bacterial spillage has been disinfected the area can be mopped up and washed with
water. Any other spill sites can also be washed with water.
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.