LS319 Lactose in Dairy Products

Faculty of Health and Life Sciences

COSHH RISK ASSESSMENT FORM

Activity Title: Type of activity: Location of activity: Lactose practical 1 and 2 Undergraduate practical 1H4/5 or 2KH1

Assessed by (+date): Endorsed by (+date): Assessment review date: A. Ainger D Lush August 2015 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 Quantity and Classification Hazard Statements Exposure (chemical, concentration (corrosive, toxic, to include exposure routes & potential Time (duration radiological or Amount in mL flammable, Biohazard health effects. / biological agents. or g then Group 1-4) From Label elements 2.2 – Hazard frequency of Including those concentration From pictogram 2.2 - Statements (H-numerical and use) created by in Mg/Ml, Please use words to description only). activity)Include %w/w, w/v, describe the pictogram. the common Molarity. See end of this form for chemical name description. from 1.1 0.1M TRIS/HCl 100mL bottle Not hazardous Not Hazardous Up to 2hrs pH 7 per 3 groups per session. (Tris(hydroxy 2 sessions methyl)amino methane) / hydrocholoric acid containing 20mM magnesium chloride

of 3 April 2013 Lactase (β 1:20 dilution Not a hazardous This substance is not classified as Up to 2hrs galactosidase) 2mL bottle substance or mixture dangerous according to Directive per session. in per group according to 67/548/EEC. 2 sessions 0.1M Tris/HCl Regulation (EC) No. buffer pH 7 + 1272/2008. 20mM magnesium chloride o-Nitrophenyl 45 mM Not hazardous Not hazardous Up to 2hrs β-D- 20mL per per session. galactopyrano group 2 sessions side (ONPG) in 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride 0.666M 0.666M 10mL Not hazardous Not hazardous Up to 2hrs in Galactose in per group one session 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride Unknown 10mL per Not hazardous Not hazardous Up to 2hrs in concentration group one session. of Galactose in 0.1M Tris/HCl buffer pH 7 containing 20mM magnesium chloride 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.

1. Induction of students into basic Health and Safety 2. Previous training of students in basic laboratory skills 3. 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. 4. Use of personal protective equipment (PPE) – lab coat and disposable gloves. 5. 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

of 3 April 2013 Indicate any specialist information relating to any of the substances in use e.g. storage, transportation, disposal, spillage, first aid Storage N/A Spillage If any of the solutions are spilled, clean up with a tissue and place in the normal waste bins. Disposal All liquids can be poured down the sink with plenty of water. First aid; N/A

Are any of the substances that you are using flammable, oxidising or explosive (name them)? No

Have you included suitable control measures above? Yes / No (delete as appropriate)

of 3 April 2013