PBS
Anti-thrombin-agarose
Hazardous Substances Policy - Assessment
CHEMICAL HAZARD AND RISK ASSESSMENT
School of Biosciences
Name of
supervisor
Dr.J.E.Turnbull
Assessor
Laine Wallace
Assessment Number*
2059
JETLW11
Date of Assessment
2/7/02
Signature
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
2
PERSONS WHO MAY BE AT RISK
List names where possible
5th Floor Biosciences Tower
Laine Wallace, Dr.J.E.Turnbull,K.Drummond,Z.Schofield,Dr.E.Yates, Dr,S Guimond,
Dr.Z.Huang, Dr.A.Powell, J.Henstock, Dr. T. Kinnunen
3
ACTIVITY ASSESSED
4
MATERIALS INVOLVED
NAME
Preparation and use of anti-thrombin affinity column.
AMOUNT
max/stored
Attach copies of data sheet(s)
HAZARD
RISK PHRASES
PBS
150ml
Irritant
R36/37/38: Irritating to eyes,
respiratory system and skin
Anti-thrombin-agarose
0.5g
N/A
N/A
NaCl
100g
Irritant
R36 : Irritating to the eyes
Heparin saccharide
0.4mg
Irritant
R36/37/38: Irritating to eyes,
respiratory system and skin
5
HAZDAT NO***
BIOSCIENCESNO***
INTENDED USE**
Give brief details and attach protocol/instructions
PBS – wash buffer
Anti-thrombin-agarose –binding medium
NaCl – to elute bound heparin
Heprin saccharide – reagent to be affinity purified
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?
PBS – risk of splashes to unprotected skin and eyes.
NaCl - risk of splashes to unprotected skin and eyes
Heparin saccharide - risk of splashes to unprotected skin and eyes
7
CONCLUSIONS ABOUT RISKS
Is level of risk acceptable? Can risk be prevented or reduced by change of substance/procedure? Are control measures necessary?
Due to the nature of some of the chemicals listed in this assessment there is a small risk . But due to the small
amounts used and as long as measures laid out in section 8 are followed then the level of risk is acceptable. Non
of the chemicals used can be replaced with other “ less hazardous” reagents.
8
CONTROL MEASURES
Additional to Good Chemical Practice
Wearing of lab coat, chemical resistant gloves and eye protection.
9
INSTRUCTION/TRAINING
Specify course(s) and/or special arrangements.
One to one initial training
10
MONITORING
Performance of control measures,
N/A
Personal exposure
Health Surveillance
N/A
N/A
11
WASTE DISPOSAL PROCEDURE
See School Server for Approved Procedure Document on specific Chemical Waste Disposal.
All reagents can be disposed of down the sink with copious amounts of water.
12
REVIEW
Enter the date or circumstances for review of assessment (maximum review interval 5 years)
2/7/02
13
EMERGENCY ACTION
TO CONTROL HAZARDS To stabilize situation eg spread absorbant on liquid spill; eliminate sources of ignition, etc.
Spills of listed chemicals can be mopped up with tissue and disposed of into contaminated waste bag.
Large spills can be covered in a absorbant material and swept up, placed into a contaminated waste bag for
normal incineration.
TO PROTECT PERSONNEL
Evacuation, protection for personnel involved in clean-up, Special First Aid
Contact with reagents within this protocol require immediate washing in copious amounts of water. If discomfort
persists then contact a physician.
Clean-up/decontamination
TO RENDER SITE OF EMERGENCY SAFE
Clean area with a dilute decon solution and dry thoroughly.
CONTACT
Dr.J.E.Turnbull
PHONE
47527
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.
Preparationof Anti-thrombin affinity column
Solutions : PBS
Equipment : 10mlplastic column.
1.
2.
3.
4.
5.
6.
Weigh out 0.5g of anti-thrombin-agarose ( Sigma A8293)
Dissolve in 10ml of PBS.
Leave for 20minutes to rehydrate.
Load into plastic column ( eg. Biorad Econcolumn ).
Wash with 50ml PBS
Store at 4C .
The column should have a capacity to bind approximately 4mg of heparin.
Affinity chromotography of heparin on anti-thrombin column.
Materials :
PBS
PBS + 0.5M NaCl
PBS + 0.75M NaCl
PBS + 1M NaCl
PBS + 1.5M Nacl
Heparin Saccharides
Equipment : AT-affinity column ( 1ml capacity approx0.8mg)
1.
2.
3.
4.
Take 0.4mg of heparin saccharide ( 20ul of 20mg.ml solution ) and add 1ml PBS.
Load onto AT-affinity column and collect eluant into a plastic tube
Re-load the eluant twice as in step 2.
Wash with 4ml PBS and collect eluant – combine with loading to give 5ml oflow affinity (PBS)
fraction.
5. Elute column with 5mlPBS + 0.5M NaCl and collect in a fresh plastic tuibe ( 0.5M fraction )
6. Repeat step 5 with 0.75M, 1M and 1.5M NaCl respectively
7. Restore column by washing with 20mlPBS. It is now ready for a further run. Store at 4 if not need
immediately.
Desalting heparin samples.
Desalt by dialysis for 16hours against distilled water using Spectrapor 1000 cut off dialysis tubing.