Mannitol

Choline bicarbonate

L-proline

Sodium chlorideHazardous Substances Policy - Assessment

CHEMICAL HAZARD AND RISK ASSESSMENT

School of Biosciences

Name of supervisor / Professor Pat Butler / Assessment Number* / 2003
Signature
Assessor / Emma J. Watkins / Date of Assessment / 08/20/2002

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 / W119 and N101
2 / PERSONS WHO MAY BE AT RISK
List names where possible / Emma J. Watkins
3 / ACTIVITY ASSESSED / Preparation of solutions for the measurement of nutrient uptake by intestinal tissue; uptake of radiochemicals by intestinal tissue; aminopeptidase-N enzyme assay.
4 / MATERIALS INVOLVED / Attach copies of data sheet(s)
NAME / AMOUNT
max/stored / HAZARD / RISK PHRASES / HAZDAT NO*** / BIOSCIENCESNO***
Mannitol / 500g / Not hazardous by CHIP / 4159
Choline bicarbonate / 100ml / No known hazards / Supplied by SIGMA
L-proline / 25g / Not hazardous by CHIP / 2320
Sodium chloride / 500g / irritant / R: 36/37/38
Irritating to eyes, respiratory system and skin / 1303
Magnesium sulphate 7-hydrate / 500g / Not hazardous by CHIP / 1211
Choline chloride / 500g / irritant / R: 36/37/38
Irritating to eyes, respiratory system and skin / Suppliedby SIGMA
Formaldehyde
38% aqueous solution
Used as 10% neutral buffered formalin
Scintillation cocktail
Biodegradable counting scintillant
Glacial acetic acid
Used as 2M solution
Sodium phosphate (monobasic)
Sodium phosphate (dibasic)
Hepes
Calcium chloride-2-
hydrate
L-alanine-p-nitroanilide / 2.5 litres
2.5 litres
16 litres
2.5 litres
100ml
250g
250g
25g
500g
100mg / Toxic
Toxic
Irritant
Corrosive
Flammable
Irritant
Irritant
Not hazardous by CHIP
Irritant / SIGMA: R23/24/25, 34, 40 43;
Toxic by inhalation; in contact with skin and if swallowed;causes burns; possible risk of irreversible effects; may cause sensitization by skin contact;
R23/24/25, 34, 40, 43:
Toxic by inhalation, in contact with skin and if swallowed; causes burns; possible risk of irreversible effects; may cause sensitization by skin contact.
R: 36 and 38
Irritating to the eyes and irritating to the skin
R: 10, 35
Flammable and causes severe burns
R: 36/37/38
Irritating to eyes, respiratory system and skin
R: 36/37/38
Irritating to eyes, respiratory system and skin
R: 36 Irritating to the eyes
c / 102
Supplied by AMERS-HAM
2
1285
641
693
1188
1312
NCS-II Tissue solubiliser
Potassium hydroxide
L-alanine-p-nitroanilide hydrochloride
p-Nitroaniline / 500ml
500g
100mg
50mg / Irritant and Flammable
Corrosive
‘Hazards have not been thoroughly investigated’
Toxic
Irritant / R: 11 and 20
Highly flammable and harmful by inhalation
R: 35; 22
Causes severe burns and harmful if swallowed
May be harmful by inhalation, ingestion or skin absorption; may cause irritation.
R: 23/24/25 and 36/37/38 and 63 and 33
Toxic by inhalation, in contact with skin and if swallowed; irritating to eyes, respiratory system and skin; possible risk of harm to the unborn child; danger of cumulative effects / Supplied by AMER-SHAM
308
5 / INTENDED USE**
Give brief details and attach protocol/instructions

Uptake of glucose and proline

The gastrointestinal tract of a duck is removed and stored in ice cold Avian Ringers solution (prepared using 20 litres distilled water, 161g sodium chloride, 33.6g sodium hydrogen carbonate, 6.9g potassium chloride, 3.2389g potassium phosphate monobasic, calcium chloride-2-hydrate and 5.866g magnesium sulphate-7-hydrate).
Sections of the intestine are mounted on stainless steel rods and everted ready for 2 minutes incubation in either:
Solution (a): 8ml 50mM L-proline + 1.4µCi 14C PEG + 6µCi 3-H L-proline or
Solution (b): 8ml 50mM D-glucose + 1.4µCi 14C D-glucose + 6µCi 3-H L-glucose
Each of the above solutions contain radioisotopes (carbon-14 and tritium). The uptake of these isotopes is measured by the addition of tissue solubiliser, followed by liquid scintillation counting. Small segments of the intestine will also be removed and stored in 10% neutral buffered formalin for later analysis.

Aminopeptidase-N assay Tissue samples are diluted 8:1 with mannitol buffer (0.0596g hepes + 0.3M potassium hydroxide + 15.925g mannitol) and homogenized. 10 microlitres of the homogenized sample are diluted 1:1, again with mannitol buffer, and placed in a test tube. To this tube is added, 1ml of the assay mix (0.5mg/ml L-alanine-p-nitroanilide + 0.2M phosphate buffer. After 20 minutes incubation at 40oC, the reaction is stopped by the addition of 3ml 2M glacial acetic acid. The absorbance of each of the resulting solutions is read at 384nm.

In order to construct a calibration curve, 0.25g p-nitroalinine are dissolved in 1litre distilled water. Four dilutions (1:1 with distilled water) are performed. From each of these dilutions, 1ml of the solution is mixed with 3ml glacial acetic acid. The absorbance of each of the resulting solutions is read at 384nm.

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?
p-nitroalinine poses a threat to the unborn child; there is a danger of cumulative effects. The following substances are irritating to the eyes, skin and respiratory system: sodium phosphate (monobasic and dibasic), sodium chloride, choline chloride and p-nitroalinine
10% neutral buffered formalin is toxic by inhalation and by contact with the skin; harmful if swallowed and can cause burns.
Glacial acetic acid and potassium hydroxide cause severe burns; a 2M solution of glacial acetic acid is irritating to the eyes and skin. Magnesium sulphate-7-hydrate and calcium chloride-2-hydrate may irritate the eyes; care should be taken to avoid generating dust.
7 / CONCLUSIONS ABOUT RISKS
Is level of risk acceptable? Can risk be prevented or reduced by change of substance/procedure? Are control measures necessary?
Yes, the level of risk is acceptable.
Tissue samples are only small (1-2 centimetres in length), thus only small volumes of the named solutions will be required.
8 / CONTROL MEASURES
Additional to Good Chemical Practice
Proper labelling of storage containers which should be appropriately stored and, if necessary, transported in special containers. Glacial acetic acid and tissue solubiliser should be kept well away from any sources of ignition. Protective gloves and clothing should be worn at all times to prevent personal contact with any substances that are toxic/irritant and or cause (severe) burns. The preparation of the p-nitroalinine solution for the enzyme assay,the use of tissue solubiliser, formalin and glacial acetic acid should be carried out in a fume cupboard.
Practice runs (without isotopes) will be performed for each uptake experiment to ensure that the procedure can be carried out quickly and smoothly, thereby minimizing contact time. Soap, water and paper towels should be available. Protective gloves and face mask should be worn when carrying out the homogeniastion.
For radiochemicals, see Approved Scheme No. PB/C14/2901 and PB/TRI/2901
9 / INSTRUCTION/TRAINING
Specify course(s) and/or special arrangements.
Training in the everted sleeve technique given by Dr W. H. Karasov, Wildlife Ecology Department, University of Madison, USA (November 2001).
Training in the assay of aminopeptidase-N given by Enrique Caviedes-Vidal, Wildlife Ecology Department, University of Madison, USA (November 2001).
Training in dissection techniques at Cherry Valley Farms Ltd, March 2001
Attendance on the ‘Working with Raioactivity’ course (October 2001)
10 / MONITORING
Performance of control measures,
Review every 5 years unless list of chemicals or quantities change.
Personal exposure / Health Surveillance
11 / WASTE DISPOSAL PROCEDURE
See School Server for Approved Procedure Document on specific Chemical Waste Disposal.
Small quantities of formailn can be poured down the sink with plenty of water. Formalin should be diluted so that the formaldehyde concentration is less than 1% before it is disposed of via the drains. Solutions containing small quantities of radiochemicals (from this protocol, upto 30 microcuries) can be carefully poured down the designated sink within the isotope lab.
The following substances can be disposed of in small quantities by diluting with water (until any reaction is complete) and then pouring down the sink, with plenty of water: 2M glacial acetic acid (dilute to <5% concentration), sodium phosphate (monobasic) and (dibasic), potassium hydroxide (dilute to <2%), magnesium sulphate-7-hydrate and calcium chloride-2-hydrate.
Contaminated gloves, paper towels e.t.c. (Hazardous waste for collection) can be placed in a sealed bag. This bag can then be placed inside a yellow bin liner labeled ‘clinical waste’ for collection by cleaning staff.
After the clean-up of a large spillage containing ‘flammable-extremely flammable’ substances, or ‘irritant’ (R35≥1% or R35≥5%), waste should be treated as ‘Special Waste’ and advice sought from the Health and Safety Unit.
In the event of a large spillage of aqueous radioactive material, personnel not involved in the clean-up should be kept away from the site. Personnel dealing with the spillage should check for personal injury and or contamination, mop up the spillage, place all waste in a sealed plastic bag, monitor the area for contamination, then contact the Radiation Safety Officer and or the Health and Safety Unit for advice on waste disposal.
12 / REVIEW
Enter the date or circumstances for review of assessment (maximum review interval 5 years)
Review in five years time or in the event of a change to the protocol.
13 / EMERGENCY ACTION
TO CONTROL HAZARDS / To stabilize situation eg spread absorbant on liquid spill; eliminate sources of ignition, etc.
Carry out work in the fume cupboard; eliminate sources of ignition; ensure access to water and appropriate fire extinguishers. In the event of spillage, wear protective clothing, keep personnel not involved in practical work away from the area, mop up spillages with an inert absorbant material e.g. paper towels; scoop up solid spill and place in a labeled container; avoid generating dust; afterwards, thoroughly wash the area with plenty of water; ventilate the area. Prevent materials entering drains or watercourses.
For further details see attached safety data sheets.
N.B. neutralizing a spill of 2M glacial acetic acid with sodium hydrogen carbonate, sodium carbonate or calcium carbonate will produce large amounts of carbon dioxide gas; ensure adequate ventilation.
University emergency service tel: 44444 report accidents and or incidents immediately.
TO PROTECT PERSONNEL / Evacuation, protection for personnel involved in clean-up, Special First Aid
For any of the listed solutions in the event of: SKIN CONTACT: Drench the affected area with clean running water for at least 10 minutes and until no chemical remains in contact with the skin. Remove contaminated clothing, which is not stuck to the skin, as soon as practicable after commencement of washing. If there is any injury or if skin absorption is suspected obtain medical attention.
EYE CONTACT: Flush the eye with clean running water for at least 10 minutes; obtain medical examination of the affected eye.
INGESTION: Do not make the casualty vomit; wash out the mouth with water. If the casualty is conscious, give 0.5 litre water to drink. Seek medical attention.
INHALATION: Remove the casualty from exposure; rest and keep warm. If the casualty has been seriously affected, obtain medical attention.
For full details of first aid measures, see attached safety data sheets.
TO RENDER SITE OF EMERGENCY SAFE / Clean-up/decontamination
Wearing protective clothing, gloves and eyeshield, mop up spillages and place contaminated waste in a sealed ‘hazardous waste’ labelled bag. This can be stored in the fume cupboard until collection. The area should be well ventilated and personnel not involved in cleaning up, should be kept well away from the area.
CONTACT : Norman Day TEL: 45474

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.