1 CHE-554 Biological Chemistry Lab - University of Kentucky€¦ · CHE-554 Biological Chemistry Lab ... You respect the safety of others in every laboratory operation that ... –

1

© A.-F. Miller 2014

CHE-554 Biological Chemistry Lab

Tues and Thurs, 12:30 pm CP 222 & 1:30 or 3:30 in CP 236

Bring lab coat, safety glasses, USB memory stick, notebook.

Wear sensible clothing: closed-toe shoes, no flammable

synthetics or loose items.

Be prepared in advance for each day.

COME

Watch for these techniques in the literature, and the news.

Why did you choose this course ?

Syllabus, and more, will be available at:

www/chem.uky.edu/courses/che554

Before the site is ready, handouts will be distributed through

email.

2


SAFETY in the Biological Chemistry Lab

Prof. Wei

January 2015

3


SAFETY in the Biological Chemistry Lab

Don’t work alone unless the task is genuinely routine for you.

– Tired, bored, stressed, distracted

– Don’t have the correct tools

– Other ?

When do accidents happen ?

– Attitude

– Alert

– Prepared

– Skill and experience

– Correct Equipment

– Equipment is in good condition

– Good environment (lighting, distractions).

Your safety equipment:

4


General Laboratory Safety

The biological chemistry laboratory can be a

dangerous place if:

You are not being careful and not paying attention to

what you are doing.

You do not understand what you are doing.

You do not use equipment or techniques properly.

https://www.youtube.com/watch?v=s6lOQ5_Vlok

5


Authorized Activities and Proper Conduct

It is very important that:

You follow the operating procedures suggested in the handouts,

the textbook, and in equipment manuals.

You only perform the assigned experiments.

You pay attention to the direction and warnings of your supervisor

or instructor.

You respect the safety of others in every laboratory operation that

you perform.

Establish a buddy system, and use it.

6


Most common reasons experiments fail

Errors following the protocol

– Write and diagram out an action plan for the entire experiment,

compiling a list of required reagents and equipment, IN

ADVANCE.

– Check steps off as they are completed, and WRITE DOWN

observations regarding the condition of the samples at each

step (write down the time as well).

– Keep an eye on total volumes, these will reveal when a reagent

has not been added, or been added in the wrong quantity.

– Check all solutions, before using them (eg. confirm pH).

This will be done for you.

7


Dress Code Safety glasses must be worn at all times. It is recommended that you not

wear contact lenses in the laboratory.

Minimize skin exposure. Legs and feet should be completely covered.

Long pants, skirts, dresses, etc. must cover legs and ankles. Shorts, capri

pants, tank tops, midriff shirts, muscle shirts, open-toed shoes, and

sandals are not permitted.

Wear a lab coat at all times.

Wear exam gloves at all times.

Tie back or confine long hair and loose items of clothing and jewelry.

Choose non-flammable clothing.

Turn off any distractions: cell phones etc. NEVER wear head phones in a

lab.

Do not even enter the room if you are under the influence of any disruptive

substance: alcohol, sedatives, drugs, etc.

8


Flammable Materials

Keep flames away from the area where you use

ether, acetone, methanol, ethanol or other organic

solvents.

Handle flammable solvents in a fume hood.

Do not inhale chemical vapours. They can be

deadly.

Wear non-synthetic clothing and shoes that provide

a barrier to liquid penetration (athletic shoes w.

mesh top and sides fail this test).

9


Biohazards

Your resident bacterial flora can contaminate your

experiment.

Dyes such as ethidium bromide are carcinogens

with delayed but serious toxicity.

Gloved hands are research hands. Research hands

do research, ONLY: no hair adjustments, wiping

face, use of public facilities ... (avoid using your

keyboard or cell phone).

10


Spills

All spills must be cleaned immediately (biological or

chemical).

Biologicals or chemicals spilled on the skin should

be washed off immediately by flushing with water for

15 minutes. Notify T.A. immediately.

In case of a major accident, seek medical treatment

immediately. (Also notify T.A. and instructor.)

11


Waste Disposal Dispose of all waste properly. Waste containers will be provided. If

unsure, ask your supervisor or instructor.

Fully read the labels on chemical and waste containers before

disposal.

Do not forget to close the container lid when done. For volatile

substances, do not tighten the lid.

Biohazard materials must be disposed off in appropriate bags.

Infectious Material must be disposed of in red bags.

Sharp materials such as gel cutters, razor blades, and syringe

needles much be disposed off in “sharps” container.

12


Concentrated Acids and Bases

When handling acids or bases:

Be careful to avoid splashing them on skin or clothes.

Dilute by adding the reagent to water (never add

water to the reagent).

13


Toxic Chemicals Toxic chemicals such as acrylamide, caustic solutions, cyanides,

oxalates, mercury salts, are hazardous to humans.

Never handle such chemicals such that your skin is in direct contact with them.

Never mouth-pipette anything.

Use impermeable gloves and nose mask while weighing or using these chemicals.

Wash any chemical spills off your body promptly.

Wash your hands with soap after EVERY session in lab, regardless of whether any mishap occurred.

Familiarize yourself will all reagents beforehand.

14


Safety of reagents

MSDS

Merck Manual

Web sites

http://cartwright.chem.ox.ac.uk/hsci/chemicals/aspiri

n.html

provides a simplified version. Authentic ones for

aspirin and glucose are available via the course web

site.

Also go through www.sigmaaldrich.com/united-

states

15


Safety of reagents

MSDS

– often prepared with the intension of providing legal

protection to the manufacturer, rather than practical

information for the user.

– Look at the LD50, IC50

– Know these values as they apply to a human (you !)

For dangerous materials

– Know exposure routes

– Know protective devices and protocols

– Know antidotes and first-response actions.

16


Safety Equipment

Know the location of all safety equipment

– Showers

– Eye washes

– Fire extinguishers

– First aid kit

– Evacuation plan

17


Handling Glass

Glass cuts are among the most common lab

accidents. Even the most experienced chemists are

susceptible. Please be careful.

Do not insert glass tubing into rubber stoppers or

corks.

Discard cracked and chipped glassware in glass

disposal box.

18


Reporting Accidents

Report an accident or an injury, no matter how

minor, to the instructor or laboratory supervisor

immediately. An accident report form must be

completed.

A first aid kit for minor cuts or burns is available in

the laboratory. Ask the instructor or laboratory

supervisor for help.

19


Safety Information

In Case of an Emergency

– Administer First Aid

– Stay with the injured person

Call 9-911

Important Phone Numbers:

– UK Police Dept. (7-8573)

– Environmental Health and Safety (7-3241)

– Occupational Health and Safety (7-3827)

– Hazardous Materials Management (3-6280)

Accident Report Form

– www.uky.edu/FiscalAffairs/Environmental/ohs/

Adhere to Building Emergency Evacuation Plan.

– Take note of your locker-mate’s successful evacuation.

– Stay together as a class to facilitate accounting for all members.

20 Section1 Lockers & Groups

Name Locker Group Bale, Michael Javier L28C A

Bosshammer, Samantha Leigh L28C B

Combs, Travis Allen L28C C

Do, Christy Luc L31C A

Hayden, Christopher Stephen L31C B

Hempel, Karl Willis L31C C

Holdeman, Kyle Robert L34C A

Hong, Calvin Ying L34C B

Howell, Nicholas Randolph L34C C

Huynh, Bobby L L35C A

Kelly, Thomas Francis L35C B

Lammers, Erin Elizabeth L35C C

Mattingly, Jacob Michael L38C A

Nguyen, Kim D L38C B

Phillips, Jared Allen L38C C

Sandell, Katelyn L41C A

Tran, Angela L41C B

Yu, Samuel Shung-Bon L41C C

L44C A

L44C B

L44C C

21


Section2 Lockers & Groups Name Locker Group

Lee, Sunyoung L28C A

L28C B

L28C C

Lim, Leang L31C A

L31C B

L31C C

Nagy, Tamas L L34C A

L34C B

L34C C

Voss, Gareth James L35C A

L35C B

L35C C

Fortenbery, Zachary Ryan L38C A

L38C B

L38C C

L41C A

L41C B

L41C C

L44C A

L44C B

L44C C

Documents

1 CHE-554 Biological Chemistry Lab - University of Kentucky€¦ · CHE-554 Biological Chemistry Lab ... You respect the safety of others in every laboratory operation that ... –