Faculty of Health and Life Sciences COSHH RISK ASSESSMENT FORM Activity Title: Lactose practical 1 and 2 Type of activity: Undergraduate practical Location of activity: 1H4/5 or 2KH1 Assessed by (+date): A. Ainger Endorsed by (+date): D Lush August 2015 Assessment review date: August 2016 Description of experimental procedure. (Give sufficient detail so that it is clear as to the procedure being undertaken, to include substances to be used and importantly created) Ortho-nitrophenyl-beta-D-Galactopyranoside (ONPG) is used instead of lactose as a substrate for the enzyme lactase. A yellow colour is produced which can be followed at 420nm in a spectrophotometer and record the data using a laptop. Students prepare various concentrations of ONPG for session 1 to record initial rates of the reaction with a laptop attached to a spectrophotometer. In session 2 students pipette various concentrations of galactose, which is used as an inhibitor of lactose. Students record initial rates of reaction with a laptop attached to a spectrophotometer Students have an unknown galactose solution which they have to work out the concentration of. Have you used the least hazardous substances possible in this procedure in order to eliminate or minimise the hazards to health? (✓) Yes No (If No please justify ) Substance details Substance (chemical, radiological or biological agents. Including those created by activity)Include the common chemical name from 1.1 Quantity and concentration Amount in mL or g then concentration in Mg/Ml, %w/w, w/v, Molarity. Classification (corrosive, toxic, flammable, Biohazard Group 1-4) From pictogram 2.2 - Please use words to describe the pictogram. See end of this form for description. Hazard Statements to include exposure routes & potential health effects. From Label elements 2.2 – Hazard Statements (H-numerical and description only). Exposure Time (duration / frequency of use) 0.1M TRIS/HCl pH 7 / hydrocholoric acid containing 20mM magnesium chloride 100mL bottle per 3 groups Not hazardous Not Hazardous Up to 2hrs per session. 2 sessions
Lactase (β galactosidase) in 0.1M Tris/HCl buffer pH 7 + 20mM magnesium chloride 1:20 dilution 2mL bottle per group Not a hazardous substance or mixture according to Regulation (EC) No. 1272/2008. This substance is not classified as dangerous according to Directive 67/548/EEC. Up to 2hrs per session. 2 sessions o-Nitrophenyl (ONPG) in 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride 45 mM 20mL per group Not hazardous Not hazardous Up to 2hrs per session. 2 sessions 0.666M Galactose in 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride 0.666M 10mL per group Not hazardous Not hazardous Up to 2hrs in one session Unknown concentration of Galactose in 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride 10mL per group Not hazardous Not hazardous Up to 2hrs in one session. Identify those persons at increased risk of harm (disabled, pregnancy, immuno-compromised etc) None Control measures for the activity - from Precautionary Statements 2.2 i.e.: Engineering controls e.g. use of fume cupboard etc., PPE/RPE (detail type), restricting quantity of substance used, prohibiting lone working, level of supervision, exposure monitoring, relevant SOPs. Induction of students into basic Health and Safety Previous training of students in basic laboratory skills A verbal introduction to the practical session will be delivered by a competent member of staff (academic and/or technical), during which specific risks and control measures will be highlighted to the students, including introduction of specific handling and dispensing techniques. Use of personal protective equipment (PPE) – lab coat and disposable gloves. Laboratory coat and nitrile gloves to be worn at all times. Additional special measures of control i.e. particular training in handling the substances listed or for the experimental procedure, relevant health surveillance etc None