AP® Investigation #13
Enzyme Activity – Teacher’s Guide
Kit # 36-7413
Table of Contents
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Abstract. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
General Overview . . . . . . . . . . . . . . . . . . . . . . . . 2
Recording Data. . . . . . . . . . . . . . . . . . . . . . . . . . 3
Materials Checklist. . . . . . . . . . . . . . . . . . . . . . . . 5
curriculum alignment . . . . . . . . . . . . . . . . . . . . . . 6
Learning Objectives. . . . . . . . . . . . . . . . . . . . . . . . 7
Time Requirements . . . . . . . . . . . . . . . . . . . . . . . . 7
Safety Precautions. . . . . . . . . . . . . . . . . . . . . . . . 8
Pre-Lab Preparation . . . . . . . . . . . . . . . . . . . . . . . 9
Student guide contents
Background. . . . . . . . . . . . . . . . . . . . . . . . . . 12
Part 1: Structured inquiry – establishing a baselline. . . 19
Part 2: Guided Inquiry – variables that affect the
rate of enzyme reaction. . . . . . . . . . . . . . . . . . . 22
Part 3: OPEN Inquiry – design an experiment . . . . . . . . . 25
MATERIAL SAFETY DATA SHEETS. . . . . . . . . . . . . . . . . . . . 27
**AP® and the Advanced Placement Program are registered trademarks
of the College Entrance Examination Board. The activity and materials
in this kit were developed and prepared by Ward’s Natural Science
Establishment, which bears sole responsibility for their contents.
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Enzyme Activity: Teacher’s Guide
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abstract
Chemical reactions underlie metabolism. Organisms have evolved catalytic proteins, called enzymes,
that can make the reactions more efficient by lowering the activation energy of a chemical reaction.
Catalytic efficiency of enzymes is dependent upon the precise shape of the active site in the protein
that interacts with substrates and products. The abiotic conditions of the reaction affect the rate of
enzyme-mediated conversion of substrate to product by affecting the conformation of this active site.
In this set of investigations, students will use an extract of turnips containing the enzyme peroxidase
to react with its substrate (hydrogen peroxide) and a color indicator (guaiacol) for formation of
product (O2), to estimate the rate of peroxidase activity under a variety of experimental conditions.
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general Overview
The College Board has revised the AP Biology curriculum to begin implementation in the fall of
2012. Advanced Placement (AP) is a registered trademark of the College Entrance Examination
Board. The revisions were designed to reduce the range of topics covered, to allow more depth of
study and increased conceptual understanding for students. There is a shift in laboratory emphasis
from instructor-designed demonstrations to student-designed investigations. While students may be
introduced to concepts and methods as before, it is expected that they will develop more independent
inquiry skills. Lab investigations now incorporate more student-questioning and experimental
design. To accomplish this, the College Board has decreased the minimum number of required
labs from 12 to 8 while keeping the same time requirement (25% of instruction time devoted to
laboratory study). The College Board has defined seven science practices that students must learn to
apply over the course of laboratory study. In brief, students must:
1. Use models
2. Use mathematics (quantitative skills)
3. Formulate questions
4. Plan and execute data collection strategies
5. Analyze and evaluate data
6. Explain results
7. Generalize data across domains
The College Board published 13 recommended laboratories in the spring of 2011. They can be found
at: http://advancesinap.collegeboard.org/science/biology/lab
Many of these laboratories are extensions of those described in the 12 classic labs that the College
Board has used in the past. The materials provided in this lab activity have been prepared by
Ward’s to adapt to the specifications outlined in AP Biology Investigative Labs: An Inquiry-Based
Approach (2012, The College Board). Ward’s has provided instructions and materials in the AP
Biology Investigation series that complement the descriptions in this College Board publication.
We recommend that all teachers review the College Board material as well as the instructions here
to get the best understanding of what the learning goals are. Ward’s has structured each new AP
investigation to have at least three parts: Structured, Guided, and Open Inquiry. Depending on a
teacher’s syllabus, they may choose to do all or only parts of the investigations in scheduled lab
periods.
The College Board requires that a syllabus describe how students communicate their experimental
designs and results. It is up to the teacher to define how this requirement will be met. Having
students keep a laboratory notebook is one straightforward way to do this.
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Recording Data in a Laboratory Notebook
All of the Ward’s Investigations assume that students will keep a laboratory notebook for studentdirected investigations. A brief outline of recommended practices to set up a notebook, and one
possible format, are provided below.
1. A composition book with bound pages is highly recommended. These can be found in most
stationary stores. Ward’s offers several options with pre-numbered pages (for instance, item
numbers 32-8040 and 15-8332). This prevents pages from being lost or mixed up over the
course of an experiment.
2. The title page should contain, at the minimum, the student’s name. Pages should be numbered in
succession.
3. After the title page, two to six pages should be reserved for a table of contents to be updated as
experiments are done so they are easily found.
4. All entries should be made in permanent ink. Mistakes should be crossed out with a single line
and should be initialed and dated. This clearly documents the actual sequence of events and
methods of calculation. When in doubt, over-explain. In research labs, clear documentation may
be required to audit and repeat results or obtain a patent.
5. It is good practice to permanently adhere a laboratory safety contract to the front cover of the
notebook as a constant reminder to be safe.
6. It is professional lab practice to sign and date the bottom of every page. The instructor or lab
partner can also sign and date as a witness to the veracity of the recording.
7. Any photos, data print-outs, or other type of documentation should be firmly adhered in the
notebook. It is professional practice to draw a line from the notebook page over the inserted
material to indicate that there has been no tampering with the records.
For student-directed investigations, it is expected that the student will provide an experimental plan
for the teacher to approve before beginning any experiment. The general plan format follows that of
writing a grant to fund a research project.
1. Define the question or testable hypothesis.
2. Describe the background information. Include previous experiments.
3. Describe the experimental design with controls, variables, and observations.
4. Describe the possible results and how they would be interpreted.
5. List the materials and methods to be used.
6. Note potential safety issues.
(continued on next page)
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Recording Data in a Laboratory Notebook (continued)
After the plan is approved:
7. The step-by-step procedure should be documented in the lab notebook. This includes recording
the calculations of concentrations, etc., as well as the weights and volumes used.
8. The results should be recorded (including drawings, photos, data print outs, etc.).
9. The analysis of results should be recorded.
10. Draw conclusions based on how the results compared to the predictions.
11. Limitations of the conclusions should be discussed, including thoughts about improving the
experimental design, statistical significance, and uncontrolled variables.
12. Further study direction should be considered.
The College Board encourages peer review of student investigations through both formal and
informal presentation with feedback and discussion. Assessment questions similar to those on the AP
exam might resemble the following questions, which also might arise in peer review:
•
Explain the purpose of a procedural step.
•
Identify the independent variables and the dependent variables in an experiment.
•
What results would you expect to see in the control group? The experimental group?
•
How does XXXX concept account for YYYY findings?
•
Describe a method to determine XXXX.
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Materials checklist
MATERIALS PROVIDED IN KIT
MATERIALS NEEDED BUT NOT PROVIDED
Units
per kit/
group????
Description
Test tube racks\
1
pH paper, range 1-14, Vial/100
Ice
1 pkg./12
Glass stirring rod, 6 mm x 150 mm
Beakers, 1000 mL, 150 mL, 50 mL
48??
Tubes, 15 x 125 mm
Parafilm
1
Guaiacol, 250 mL
Distilled water
24
Syringe, 2.5 mL
Timer
16
Syringe, 10 mL
Brown bottle, 500 mL or bottle wrapped in foil
1
Hydrogen peroxide, 3%, lab grade
Beakers or flasks, 500 mL
1 pkg./15
Pipet, 6”
Graduated cylinders, 20-500 mL and 1L
1
Scholar Chemistry MSDS CD-Rom
Refrigerator
3
Cheesecloth, 9 x 12 piece
Blender
1 pkg./20
Disposable pipet. 9”
Other materials as determined by students’
experimental design
1
Buffer set:
Includes envelopes of pH 2-11
(one each, for a total of 10 envelopes),
500 mL buffer
Personal protective equipment (gloves, apron,
safety glasses)
1
Instructions (this booklet)
OPTIONAL MATERIALS (NOT PROVIDED)
Light probe or spectrophotometer
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This lab activity is aligned with the 2012 AP Biology Curriculum (registered trademark of the College Board).
Listed below are the aligned Content Areas (Big Ideas and Enduring Understandings), the Science Practices, and the
Learning Objectives of the lab as described in AP Biology Investigative Labs: An Inquiry Approach (2012). This is a
publication of the College Board that can be found at http://advancesinap.collegeboard.org/science/biology/lab.
Curriculum alignment
Big Ideas
‹ Big Idea 4: Biological systems interact, and these interactions possess complex properties
Also connects to:
‹ Big Idea 2: Biological systems utilize energy and molecular building blocks to grow, to
reproduce, and to maintain homeostasis.
Enduring Understandings
‹ 2.D.1: All biological systems from cells and organisms to populations, communities and
ecosystems are affected by complex biotic and abiotic interactions involving exchange of matter
and free energy.
‹ 4A1: The subcomponents of biological molecules and their sequence determine the properties of
that molecule.
‹ 4B1: Interactions between molecules affect their structure and function.
Science Practices
‹
‹
‹
‹
don’t see any science practices listed in
word doc.
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learning objectives
‹ The student is able to design a plan for collecting data to show that all biological systems are
affected by complex biotic and abiotic interactions (2D1 & SP 4.2, SP 7.2).
‹ The student is able to use models to predict and justify that changes in the subcomponents of a
biological polymer affect the functionality of the molecule (4A1 & SP 6.1, SP 6.4).
‹ The student is able to analyze data to identify how molecular interactions affect structure and
function (4B1 & SP 5.1).
Time Requirements
Structured Inquiry
Guided Inquiry
Open Inquiry
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10 minutes
45 minutes
45 minutes
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Safety Precautions
Lab-Specific Safety
‹ Guaiacol is a poison and flammable. Review the Material Safety Data Sheet (MSDS) for this
chemical.
‹ Remind students to use proper pipeting techniques. Never mouth pipet
General Safety
‹ The teacher should be familiar with safety practices and regulations in their school (district and
state). The teacher should know what needs to be treated as hazardous waste and how to properly
dispose of non-hazardous chemicals or biological material.
‹ Consider establishing a safety contract that students and their parents must read and sign off on.
This is a good way to identify students with allergies to things like latex so that you (and they)
will be reminded of what particular things may be risks to individuals. A good practice is to
include a copy of this contract in the student lab book (glued to the inside cover).
‹ Students should know where all emergency equipment (safety shower, eyewash station, fire
extinguisher, fire blanket, first aid kit etc.) is located.
‹ Make sure students remove all dangling jewelry and tie back long hair before they begin.
‹ Remind students to read all instructions, Material Safety Data Sheets (MSDSs) and live care
sheets before starting the lab activities and to ask questions about safety and safe laboratory
procedures. Appropriate MSDSs and live care sheets can be found on the last pages of this
booklet. Additionally, the most updated versions of these resources can be found at www.wardsci.
com, under Living Materials
http://wardsci.com/article.asp?ai=1346. (Note that in this particular lab, there are no live
materials that require a live care sheet.
‹ In student directed investigations, make sure that collecting safety information (like MSDSs) is
part of the experimental proposal.
‹ As general laboratory practice, it is recommended that students wear proper protective
equipment, such as gloves, safety goggles, and a lab apron.
At end of lab:
‹ All laboratory bench tops should be wiped down with a 20% bleach solution or disinfectant to
ensure cleanliness.
‹ Remind students to wash their hands thoroughly with soap and water before leaving the
laboratory.
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Kit # 36-7413
Pre-Laboratory Preparation
Notes
1. Make copies of Student Guide.
Copy pages __ to __ of the student copymaster prior to starting
class.
2. Dilute hydrogen peroxide solution
‹
NOTE: This can be made a day ahead and stored overnight
in the refrigerator protected from light in either a brown
bottle or foil wrapped bottle.
Prepare 2 liters of the dilute solution by adding 66 mL of the
stock 3% hydrogen peroxide solution to 2 liters of distilled
water. Dispense into 8 bottles and label this solution 0.1% H2O2.
‹
NOTE: Be extremely precise in your measurements of all
reagents to avoid erroneous results.
3. Prepare pH buffer solutions.
‹
NOTE: This can be prepared a day ahead and stored covered
overnight at room temperature.
a) Label each of 6 large beakers so that there is one beaker per
pH buffer envelope.
b) Measure 500 mL of distilled water and pour 500 mL of
water into each beaker.
c) Empty one entire powder-filled envelope over into the filled
beaker, and label the beaker that matches the pH indicated
on the envelope. Mix by stirring the powder into solution.
d) Measure the pH of the buffer solution to confirm that the
desired pH is achieved.
‹
OPTIONAL: Dispense buffer into 8 bottles, one for each lab
group.
4. Prepare stock indicator solution.
‹
NOTE: This can be prepared the day before the experiment
and stored at 4 °C, protected from light.
‹
NOTE: Keep the guaiacol solution on ice in a designated
area throughout the course of the lab to maintain stability.
Dilute 1.5 mL guaiacol in 500 mL water in either a brown bottle
or a bottle covered with aluminum foil.
(continued on next page)
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Pre-Laboratory Preparation (Continued)
Notes
5. Prepare stock enzyme solution.
‹
NOTE: The turnip peroxidase solution can be prepared the
day before the experiment. The solution is stable for up to
one week if stored at 4 °C and protected from light.
The enzyme solution is an extract made from a fresh turnip
(purple topped or other available at the grocery store). Weigh
out 25 g of a peeled turnip. Place into a blender and cover
with 400 mL of cold distilled water. Blend on high speed for
1 minute. Filter the puree through three layers of cheesecloth.
Discard the cheesecloth with the solids. Pour the solution into
either in a brown bottle or a bottle covered with aluminum foil,
and store in refrigerator, protected from light. Let any starch
precipitate settle to bottom of bottle.
‹
TIP: It is always best to use distilled water, but tap water
may be used as well.
‹
NOTE: Keep the turnip peroxidase solution on ice in a
designated area throughout the course of the lab to keep the
enzyme activity stable.
6. Prepare the color palette.
‹
NOTE: Prepare the standardized color palette the day before
the lab activity to ensure the reactions have maximized
the production of the enzyme-substrate reaction product.
This will represent your “brownest” sample (10). This
color palette will be used by the students to approximately
quantify the reaction rate of the enzyme-substrate reaction.
Store in refrigerator overnight.
a. Prepare Enzyme-substrate mixture
(enough to make one color palette)
In a 150 mL beaker add each of the following:
15 mL stock turnip peroxidase solution
30 mL 0.1% H2O2 stock
and
15 mL stock indicator – guaiacol solution
‹
NOTE: Be sure to change the disposable pipets in between
solutions so there is no carry over.
(continued on next page)
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Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Pre-Laboratory Preparation (Continued)
Notes
6. Prepare the color palette (continued)
b. Label 11 test tubes with the numbers 0-10 respectively.
Using the chart below, fill the respective tube with the
specific amounts of each reagent:
Test
Tube
Number
Final Solution
Quantity Desired
(mL)
Concentration
of
Final Solution
(%)
mL of
EnzymeSubstrate
Mixture
mL of
Distilled Water
10
9
8
7
6
5
4
3
2
1
0
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7.5
100
90
80
70
60
50
40
30
20
10
0
7.50
6.75
6.00
5.25
4.50
3.75
3.00
2.25
1.50
0.75
0.00
0.00
0.75
1.50.
2.25
3.00
3.75
4.50
5.25
1.50
6.75
7.50
‹
OPTIONAL: The color palette can be quantified using a
spectrophotometer to measure absorbance at a wavelength
of 436 nm.
(continued on next page)
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Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Background
OBJEcTIVES
‹ Design a plan for collecting
data to show that all biological
systems are affected by complex
biotic and abiotic interactions.
‹ Use models to predict and
justify that changes in the
subcomponents of a biological
polymer affect the functionality
of the molecule.
‹ Analyze data to identify how
molecular interactions affect
structure and function.
Enzymes, Substrates and Products
Chemical reactions underlie metabolism. Organisms have evolved
catalytic proteins, called enzymes, that can make the reactions more
efficient by lowering the activation energy of a chemical reaction.
Catalytic efficiency of enzymes is dependent upon the precise shape
(conformation) of the active site in the protein that interacts with
substrates and products.
When the enzyme is reacting with the substrate, a complex is formed.
This interaction can be expressed as:
Figure 1
In the induced fit model of enzymatic activity, enzymes change shape
after binding to a substrate, improving the “fit” between the enzyme
and the substrate.
That fit between the enzyme and the substrate is responsible for
lowering the activation energy required to transform substrates into
products. The graph below illustrates the differences between the
amounts of activation energy (Ea) of an uncatalyzed reaction and the
lower activation of an enzyme-catalyzed reaction. Note that there is no
difference in the final amount of free energy (Δ G).
Figure 2: Course of Reaction
(continued on next page)
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Enzyme Activity: Teacher’s Guide
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Background (COntinued)
Notes
The efficiency of an enzyme in facilitating the chemical reaction
that transforms substrates to products is changed by conditions that
affect the shape of the active site. Optimal enzyme efficiency requires
optimal environmental conditions, such as pH and temperature.
Raising the temperature of any substance will cause a rise in its
average kinetic energy, because the heat in any given system is
expressed as molecular motion. The more heat that is added, the more
molecular motion will occur. As a general rule, an increase of 10 °C
doubles the rate of most chemical reactions. Enzymes are proteins,
however, and so are subject to heat-induced alteration of their shape or
tertiary structure. The protein has become denatured when the tertiary
structure is altered so much that it becomes permanently inactive. The relative acidity or alkalinity of an enzyme’s surroundings also
influences its activity through a similar mechanism as temperature
– by altering the enzyme’s interaction with substrates through
conformational changes in the active site. The pH range that
supports an enzyme’s optimal activity is associated with the natural
environment in which it evolved. The ionization state of the R groups
of the amino acids that make up the protein are affected by the pH of
the environment. Therefore, the charge associated with an R group
in an active site will affect how efficiently the enzyme interacts with
substrates and products. Some enzymes will not be optimally active
until negative charges are neutralized or, conversely, until R groups
become charged. Two examples are illustrated in the graph below.
Pepsin is a hydrolytic enzyme that is required to be active in the acidic
environment of the stomach. Trypsin is hydrolytic intestinal enzyme
with an optimal enzymatic activity in the slightly basic range that is
produced in the pancreas.
Figure 3
(continued on next page)
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Enzyme Activity: Teacher’s Guide
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Background (COntinued)
Notes
Enzyme-facilitated reactions follow the Law of Mass Action: the
direction taken by an enzyme-catalyzed reaction is directly dependent
on the relative concentration of enzyme, substrate, and product.
When there is a great deal of substrate and little product, the reaction
will form more products. Conversely, when there is a great deal of
product, the reaction may form more substrate. However, here are
two situations where the Law of Mass Action is circumvented. When
the product is immediately metabolized or transported away from the
enzyme, the product concentration does not rise. In highly exergonic
reactions, the product has little free energy and the opposite reaction
requires a large, if not unachievable, amount of energy to reverse the
process.
Figure 4
Note that in the above graph, the initial reaction is very steep. As more
product forms, the rate of formation levels off as an equilibrated ratio
of substrate and product is produced.
Enzymes are often tightly bound to a prosthetic group (cofactor),
which is either a metal ion, an organic molecule/metal ion complex,
or a small organic molecule (coenzyme). Metal ions such as Fe+3
and Zn+2 are generally involved in reactions which require electron
removal from a substrate or which can electrically bond an enzyme
to a substrate. Coenzymes have a much more varied role. Some are
not tightly bound and can move from enzyme to enzyme, transferring
electrons or protons. Some alter substrates to better fit with the
enzyme. Still others, bound into membranes, are essential to the
energy conversion reactions of photosynthesis and respiration. Many
coenzymes must be taken in by animals and are not synthesized; these
are collectively referred to as vitamins.
(continued on next page)
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Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Background (COntinued)
Notes
Competitive Inhibitors
Molecules that bind to the active site and compete with the substrate
are called competitive inhibitors. Note that the inhibitor is similar
enough to the enzyme that it can fit in the active site, but it is not
the same as the substrate. The inhibitor binds the enzyme, therefore
blocking substrates from binding, and no product is produced.
Competitive inhibition is reversible and behaves the same as an
enzyme–substrate complex, with constant binding and unbinding of
the inhibitor due to the Law of Mass Action. If the concentration of the
inhibitor is large enough, the reaction with substrate will slow down;
otherwise, the inhibitor has little effect. Increasing the concentration
of the substrate can overcome the effect of a competitive inhibitor.
The figure below illustrates the effect of a competitive inhibitor on
an enzyme. The reaction without an inhibitor proceeds very rapidly,
whereas the reaction that is competitively inhibited proceeds more
slowly.
Figure 5: Substrate Concentration
Unlike competitive inhibitors, noncompetitive inhibitors bind to a
region of the enzyme other than the active site, causing a shape change
in the enzyme that will impair the function of the active site. Since
there is no competition for the active site, the Law of Mass Action will
not come into play in this situation, and a buildup of substrate will not
make a difference in accelerating the reaction.
(continued on next page)
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Enzyme Activity: Teacher’s Guide
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Background (COntinued)
Notes
Figure 6
More complex enzymes that have quaternary structures contain sites
in addition to the active sites which are called effector bonds. The
effector renders the enzyme inactive only while it is on the enzyme.
This is a common mechanism of control in metabolic pathways. In
the pathways, enzymes exist in two forms: active and passive. The
active form is rendered inactive by an effector, often a product of a
later enzyme reaction, as illustrated in the diagram below. When the
products of a metabolic pathway inhibit an earlier step in the pathway,
it is referred to as feedback inhibition, or negative feedback.
Figure 7???
Not all effectors inhibit an enzyme. In allosteric promotion, the effector
activates an inactive enzyme. Both of these regulatory mechanisms are
extremely effective and can work together, allowing the cell to store
enzymes in both their active and inactive forms. In allosteric inhibition,
if a product down the metabolic line begins to build up, it is not to the
cell’s advantage to continue to make it. The product itself will “turn
down” the reaction until most of the product has been metabolized. In
allosteric promotion, that same product (or a different one) will activate
an enzyme to begin reaction with a substrate.
(continued on next page)
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Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Is this number reference
correct?
Background (COntinued)
Notes
The Role of Turnip Peroxidase
Hydrogen peroxide is a highly reactive oxidizing agent that is
produced in most cells, formed spontaneously as metabolic waste.
The enzyme turnip peroxidase is found in a cell organelle called
the peroxisome that serves to sequester and break down hydrogen
peroxide into relatively inert water and oxygen gas, preventing
chemical damage that would be caused by free, reactive peroxide. Turnip cells use turnip peroxidase to break down hydrogen peroxide
into water and oxygen (Figure 6). How efficiently the enzymes are
able perform this reaction is influenced by abiotic and biotic factors.
The rate of enzymatic activity is determined by how fast 1 unit of
enzyme can convert 1µM of hydrogen peroxide to oxygen. In this lab
activity, you will perform an enzymatic activity assay in which you
will assess the amount of oxygen produced by comparing the color
development of the reaction to a color palette.
turnip peroxidase
2 H2O2
Substrate
Enzyme
2 H 2O + O 2
Products
Figure 7?? 8??
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 17
Enzyme Activity: Teacher’s Guide
Notes
Kit # 36-7413
Safety Precautions
‹ As general safe laboratory practice, it is recommended that you
wear proper protective equipment, such as gloves, safety goggles,
and a lab apron.
‹ As general lab practice, read the lab through completely before
starting, including any Material Safety Data Sheets (MSDSs) and
live materials care sheets at the end of this booklet as well as any
appropriate MSDSs for any additional substances you would like
to test. One of the best sources is the vendor for the material. For
example, when purchased at Wards, searching for the chemical on
the Ward’s website will direct you to a link for the MSDS. (Note:
There are no live materials care sheets included in this particular
lab.)
At the end of the lab:
‹ All laboratory bench tops should be wiped down with a 20%
bleach solution or disinfectant to ensure cleanliness.
‹ Wash your hands thoroughly with soap and water before leaving
the laboratory.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 18
Enzyme Activity: Teacher’s Guide
Procedure
Tips
‹ When performing this lab
activity, all data should be
recorded in a lab notebook. You
will need to construct your own
data tables, where appropriate,
in order to accurately capture
the data from the investigation.
‹ If directed to do so by your
teacher, this part of the lab may
be done at the same time as Part
2 of the lab.
Kit # 36-7413
Part 1 – Structured INQUIRY:
establishing a baseline
MATERIALS needed PER LAB GROUP
q
q
q
q
q
q
q
q
q
q
2
1
1
5
1
1 mL
1 mL
2 mL
4 mL
1 mL syringes
2.5 mL syringe
10 mL syringe
15 x 125 mm tubes
Disposable 9” transfer pipet
Guaiacol
Turnip peroxidase solution
Dilute hydrogen peroxide
pH 7 buffer solution
Timer
Shared Materials
q 1 box of Parafilm
Part 1 – PROCEDURE: Structured inquiry
1. Label the test tubes and syringes, as follows:
2.5 mL syringe labeled ‘E’ for enzyme – turnip peroxidase
solution.
2.5 mL syringe labeled ‘P’ for product as represented by
indicator . Guaiacol reacts with free O2 (product) to form brown
color.
10 mL syringe labeled ‘NB’ for neutral buffer – pH 7.
2.5 mL syringe labeled ‘S’ for substrate – 0.1% H2O2
Test tube labeled ‘SPNB’ for mixture A
Test tube labeled ‘ENB’ for mixture B
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 19
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
PROCEDURE – Part 1: Structured Inquiry (continued)
Notes
2. Fill and prepare the labeled syringes and tubes with the
appropriate solutions (provided).
a. Dispense the following reagents in “Tube SPNB”:
2 mL of ‘S’ substrate
1 mL of ‘P’ indicator for product
1 mL of ‘NB’ neutral buffer
b. Cap tube ‘SPNB’ with parafilm and gently invert two times
to mix.
c. Dispense the following reagents in “Tube ENB”:
1 mL ‘E’ enzyme (turnip peroxidase)
3 mL‘NB’ neutral buffer
d. Cap tube ‘ENB’ with parafilm and gently invert two times
to mix.
e. Using a disposable transfer pipet, transfer the mixture from
Tube ‘SPNB’ into Tube ‘ENB’.
f.
Cap with parafilm and invert two times to mix.
3. Make observations and record data.
a. Using the color palette provided by your instructor,
immediately observe and compare the color of your
reaction to the color palette and record the tube # (color)
of the mixture over time. This data should be recorded
in 1-minute intervals for a total incubation period of five
minutes.
‹
OPTIONAL: If the class has access to a probe to measure
the transmittance of light or to a spectrophotometer, more
quantitative results with better resolving power can be
generated by measuring the changes in the transmittance or
absorbance of light.
b. Plot the increase in color intensity (product formation)
relative to your color palette over the 5-minute interval and
calculate the rate of enzymatic reaction under the baseline
conditions of this experiment.
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 20
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
PROCEDURE – Part 1: Structured Inquiry (continued)
Notes
3. Make observations and record data (continued)
‹
Note: The color palette represents a range of indicator color
that increases 10% between tubes 1-10. Tube 0 represents
no indicator, no color.
‹
NOTE: The color intensity is used as way to quantify
the amount of oxygen that is produced in the enzymatic
reaction. The brown color is produced when the guaiacol
reacts with oxygen (product of the enzyme substrate
reaction). Therefore, the more intense the color, the more
oxygen is produced in the reaction.
4. Set your labeled syringes aside for use in the following parts of
this investigation.
‹
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
NOTE: Do not throw away your labeled syringes.
You will need them for the remaining parts of this
investigation.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 21
Enzyme Activity: Teacher’s Guide
Procedure
TipS
‹ When performing this lab
activity, all data should be
recorded in a lab notebook. You
will need to construct your own
data tables, where appropriate,
in order to accurately capture
the data from the investigation.
‹ If directed to do so by your
teacher, this part of the lab may
be done at the same time as Part
1 of the lab.
Kit # 36-7413
Part 2 – GUIDED INQUIRY: Variables that affect
the rate of enzyme reaction
MATERIALS needed PER LAB GROUP
q
q
q
q
q
q
12
3
1
6 mL 6 mL 12 mL 15 x 125 mm tubes
2.5 mL syringes (from Part 1) Jana, please note...
10 mL syringe (from Part 1)
add p, S, etc.
‘P’ product guaiacol
‘E’ enzyme turnip peroxidase solution
‘S’ substrate dilute hydrogen peroxide (0.1% H2O2)
Shared Materials
q
q
q
q
q
q
q
1
‘NB’ 10 mL Syringe to dispense all buffer solutions
500 mL pH 3 Buffer Solution
500 mL pH 5 Buffer Solution
500 mL pH 6 Buffer Solution
500 mL pH 7 Buffer Solution
500 mL pH 8 Buffer Solution
500 mL pH 10 Buffer Solution
Part 2 – PROCEDURE: guided inquiry
1. In your laboratory notebook or sheet, record the baseline rate
(from Part 1???).
2. Label twelve 13 x 100 mm tubes 1 through 12, respectively.
To simplify the process, pair the tubes according to the chart
below:
Tube
Tube/pH
‹
1
3
2
5
4
6
9
7
11
8
12
10
NOTE: All of the pH tubes will contain the respective pH
buffer solution, in case the tubes get mixed up.
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 22
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
PROCEDURE – Part 2: GUIDed Inquiry (continued)
Notes
3.
In each of tubes 1, 2, 4, 9, 11, and 12, dispense:
2 mL ‘S’ substrate dilute hydrogen peroxide using ‘S’ syringe
1 mL‘P’ product indicator guaiacol using ‘P’ syringe
1 mL ‘NB’ neutral buffer pH 7
‹
NOTE: All of these tubes contain the substrate.
4. In tube 3 dispense the following volumes of reagents:
1 mL of turnip peroxidase solution, using the syringe labeled ‘E’
3 mL of pH 3 solution, using the rinsed ‘NB’ syringe
5. Repeat step 4 for tubes 5, 6, 7, 8, and 10 with their respective
pH buffers.
6. Using a disposable 9” transfer pipet, combine the reagents from
tube 1 with the reagents in tube/pH 3.
7. Observe the enzyme reaction mixtures every minute for
5 minutes by comparing to the color palette (or optional
measurement of absorbance/transmittance). Record your
observations in your laboratory notebook.
8. Refer back to the tube pairing chart and mix the remaining pairs
of tubes. Repeat Steps 6 and 7 for the remaining pairs of tubes.
9. Calculate the rate of reaction for each tube as described in Part
1. In your laboratory notebook, graph your rate results relative
to pH.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 23
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Part 2 assessment questions
1. Based on the graph and the overall slope of the line, what can you conclude about the effect of pH on
reaction rate? Why do you think that occurs?
The turnip peroxidase enzyme is most effective at the pH of 7. As the pH moves away from neutral,
the conformation of the protein impairs its enzymatic activity.
2. What happens to turnip peroxidase during and after the reaction?
Turnip peroxidase remains the same throughout the whole reaction and is present once the reaction
stops. It is not considered a product of the reaction as it never changes composition, although it may
lose activity due to conformational changes.
3. What would the reaction look like if you left parts of the reaction out of the mix?
If the enzyme was left out, the reaction would be very slow, perhaps speeded with heating.
If the substrate was left out, you might expect no reaction, however, there might be some substrate
present in the turnip extract, but at a much lower concentration (would slow the reaction).
If the indicator was left out, the reaction would occur but we would not be able to detect the
formation of product. Since the product reacts with the indicator, the product may build up faster
without the indicator, possibly reaching a lower equilibrium rate.
4. What other factors may influence enzyme activity (rate of reaction)?
Temperature, starting substrate concentration, cofactors in other enzymes, inhibitor concentrations,
etc.
5. Peroxidase breaks down hydrogen peroxide. What other types of enzymes might be needed in an
organism?
Enzymes that build or break chemical bonds of biological molecules like proteins, lipids,
carbohydrates, and nucleic acids. Proteases, lipases, glycolytic enzymes, DNA polymerase, fatty acid
synthase, etc.
6. You investigated peroxidase from a turnip. How might the activity of peroxidase from a mammal be
different?
Optimal temperature would be closer to body temperature, pH optima may be different. The
sequence of the protein would also be likely to be different – but the peptides forming the active site
might be conserved during evolution.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 24
Enzyme Activity: Teacher’s Guide
EXPERIMENT
DESIGN Tips
The College Board encourages peer
review of student investigations
through both formal and informal
presentation with feedback and
discussion. Assessment questions
similar to those on the AP exam
might resemble the following questions, which also might arise in peer
review:
‹ Explain the purpose of a
procedural step.
‹ Identify the independent
variables and the dependent
variables in an experiment.
‹ What results would you expect
to see in the control group? The
experimental group?
‹ How does XXXX concept
account for YYYY findings?
‹ Describe a method to determine
XXXX.
Kit # 36-7413
Part 3: Enzyme Activity
open inquiry: design an experiment
What questions occurred to you as you completed your studies of
peroxidase activity? Now that you are familiar with enzymatic activity
and ways to assess that activity, design an experiment to investigate
one of your questions. Questions may involve assessing a range of
abiotic factor effects on peroxidase activity, modeling the kinetics of
enzyme activity, comparing protein sequence or optimal conditions
for peroxidases from different sources, testing potential chemical
inhibitors of enzyme activity, comparing sensitivities of different kinds
of enzymes to the same abiotic factors, or identifying indicators for
different kinds of enzyme activity.
Before starting your experiment, plan your investigation in your lab
notebook. Have your teacher check over and initial your experiment
design. Once your design is approved, investigate your hypothesis.
Be sure to record all observations and data in your laboratory sheet or
notebook.
Use the following steps when designing your experiment.
1. Define the question or testable hypothesis.
2. Describe the background information. Include previous
experiments.
3. Describe the experimental design with controls, variables, and
observations.
4. Describe the possible results and how they would be interpreted.
5. List the materials and methods to be used.
6. Note potential safety issues.
After the plan is approved by your teacher:
7. The step by step procedure should be documented in the
lab notebook. This includes recording the calculations of
concentrations, etc. as well as the actual weights and volumes
used.
8. The results should be recorded (including drawings, photos, data
print outs).
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 25
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Part 3: open inquiry (continued)
Notes
9. The analysis of results should be recorded.
10. Draw conclusions based on how the results compared to the
predictions.
11. Limitations of the conclusions should be discussed, including
thoughts about improving the experimental design, statistical
significance and uncontrolled variables.
12. Further study direction should be considered.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 26
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 2.00
MSDS # 126.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 2.00
Synonyms/General Names: pH 2 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder is a body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
1
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Section 4:
First Aid Measures
Sulphamic Acid (5329-14-6) 30 to 40%
Eyes:
Skin:
Ingestion:
Inhalation:
Potassium Biphthalate ( 877-24-7) 60 to 70%
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: Buffer pH 2 capsule: OSHA PEL:
N/A, ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 27
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 126.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 2.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
2.0, acidic.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 2 Buffer: LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 28
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 3.00
MSDS # 127.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 3.00
Synonyms/General Names: pH 3 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder is a body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
1
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Section 4:
First Aid Measures
Sulphamic Acid (5329-14-6) 20 to 30%
Eyes:
Skin:
Ingestion:
Inhalation:
Potassium Biphthalate ( 877-24-7) 70 to 80%
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: Buffer pH 3 capsule: OSHA PEL:
N/A, ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
1/20/2012
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 29
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 127.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 3.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
3.0, acidic.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 3 Buffer: LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 30
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 4.00
MSDS # 128.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 4.00
Synonyms/General Names: pH 4 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Potassium Biphthalate ( 877-24-7) 100%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: Potassium Biphthalate: OSHA
PEL: N/A, ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 31
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 128.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 4.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Section 12:
Ecological Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
4.0, acidic.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
Potassium Biphthalate : LD50 [oral, rabbit];3200 mg/kg; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 32
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 130.30
Section 1:
Page 1 of 2
Buffer Capsule, pH 5.00
Product and Company Identification
Buffer Capsule, pH 5.00
Synonyms/General Names: pH 5 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Potassium Biphthalate ( 877-24-7), 75 to 85%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Sodium Phosphate Dibasic, (7558-79-4), 15 to 25%
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: Potassium Biphthalate: OSHA
PEL: N/A, ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 33
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 131.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 6.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Section 12:
Ecological Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
6.0, acidic.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 6 buffer capsule : LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Section 14:
Transport Information
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 34
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 7.00
MSDS # 132.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 7.00
Synonyms/General Names: pH 7 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Sodium Phosphate Dibasic, (7558-79-4), 60 to 70%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Potassium Phosphate monobasic (7778-77-0), 30 to 40%
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: pH 7 Buffer: OSHA PEL: N/A,
ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
1/20/2012
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 35
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 132.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 7.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
7.0, neutral.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 7 buffer capsule : LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 36
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 8.00
MSDS # 134.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 8.00
Synonyms/General Names: pH 8 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Sodium Phosphate Dibasic, (7558-79-4), 80 to 90%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Potassium Phosphate monobasic (7778-77-0), 10 to 20%
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: pH 8 Buffer: OSHA PEL: N/A,
ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 37
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 134.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 8.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Section 12:
Ecological Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
8.0, neutral.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 8 buffer capsule : LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 38
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 9.00
MSDS # 135.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 9.00
Synonyms/General Names: pH 9 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Section 4:
First Aid Measures
Sodium Carbonate, (497-19-8), 20 to 30%
Eyes:
Skin:
Ingestion:
Inhalation:
Sodium Bicarbonate (144-55-8), 70 to 80%
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: pH 9 Buffer: OSHA PEL: N/A,
ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 39
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 135.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 9.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
9.0, neutral.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 9 buffer capsule : LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 40
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Buffer Capsule, pH 10.00
MSDS # 122.30
Section 1:
Page 1 of 2
Product and Company Identification
Buffer Capsule, pH 10.00
Synonyms/General Names: pH 10 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Section 4:
First Aid Measures
Sodium Carbonate, (497-19-8), 50 to 70%
Eyes:
Skin:
Ingestion:
Inhalation:
Sodium Bicarbonate (144-55-8), 30 to 50%
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: pH 10 Buffer: OSHA PEL: N/A,
ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 41
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 42
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 124.30
Section 1:
Page 1 of 2
Buffer Capsule, pH 11.00
Product and Company Identification
Buffer Capsule, pH 11.00
Synonyms/General Names: pH 11 Buffer Capsule
Product Use: For educational use only
Manufacturer: Micro Essential Laboratory Inc., Brooklyn, NY 11210
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Capsule containing white powder; no odor.
Hazards Identification
CAUTION! Powder may be a slight body tissue irritant.
Target organs: None known
HMIS (0 to 4)
Health
0
Fire Hazard
0
Reactivity 0
This material is not considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200) if used properly.
Section 3:
Composition / Information on Ingredients
Sodium Phosphate, tribasic, (7601-54-9), 20 to 30%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Sodium Bicarbonate (144-55-8), 70 to 80%
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
Accidental Release Measures
Section 7:
Handling and Storage
0
0
0
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Sweep up spill and place in sealed bag or container for disposal. Wash spill area after pickup is complete. See Section
13 for disposal information.
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: pH 11 Buffer: OSHA PEL: N/A,
ACGIH: TLV: N/A, STEL: N/A.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
1/20/2012
250-7462 v.5/12
Page 43
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 124.30
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Buffer Capsule, pH 11.00
N/A.
N/A.
N/A
N/A
N/A
N/A.
N/A.
N/A.
N/A.
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Section 10:
Stability and Reactivity
Section 11:
Toxicology Information
Scholar Chemistry
Capsule with white solid.
No odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
11.0, neutral.
N/A.
N/A.
N/A = Not available or applicable
Avoid heat and moisture.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Acids, alkalis,
Shelf life: Indefinite if stored properly.
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
pH 11 buffer capsule : LD50 [oral, rabbit]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Ecological impact has not been determined.
Section 13:
Disposal Considerations
Section 14:
Transport Information
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Current Issue Date: January 20, 2012
Other Information
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2009, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/20/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 44
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Guaiacol
MSDS # 320.00
Section 1:
Page 1 of 2
Product and Company Identification
Guaiacol
Synonyms/General Names: 2-Methoxyphenol
Product Use: For educational use only
Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Hazards Identification
Pale, yellow liquid or solid [melting point is RT], characteristic odor.
CAUTION! Combustible liquid, keep away from all ignition sources. Moderately toxic by
ingestion and skin absorption. Body tissue irritant.
Target organs: Skin
HMIS (0 to 4)
Health
2
Fire Hazard
1
Reactivity 0
This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).
Section 3:
Composition / Information on Ingredients
Guaiacol (90-05-1), 98%
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Aspiration hazard. Rinse mouth with cold water. Give victim 1-2 tbsp of
activated charcoal mixed with 8 oz water.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Combustible Liquid. When heated to decomposition, emits acrid fumes
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool container
with water spray. Material is not sensitive to mechanical impact. Material is sensitive to static discharge.
Section 6:
2
2
1
Accidental Release Measures
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a
sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.
Section 7:
Handling and Storage
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in Flammable Area [Red Storage] with other flammable materials and away from any strong oxidizers. Store in a
dedicated flammables cabinet. Store in a cool, dry, well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Use NIOSH-approved respirator with an acid/organic
cartridge. Exposure guidelines: Guaiacol: OSHA PEL: N/A, ACGIH TLV: N/A, STEL: N/A.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
12/21/2011
250-7462 v.5/12
Page 45
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 320.00
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Guaiacol
Scholar Chemistry
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
CH3OC6H4OH.
124.14.
1.129 g/mL @ 20°C.
4.27.
28°C.
205°C.
N/A.
82°C (179°F) CC.
385°C (725°F).
Section 10:
Pale, yellow liquid.
Characteristic odor.
N/A.
Alcohol, ether, acetic acid.
N/A . (Butyl acetate = 1).
N/A. (log POW).
N/A.
3.3%.
19 %.
N/A = Not available or applicable
Stability and Reactivity
Avoid heat and ignition sources.
Stability: Stable under normal conditions of use.
Incompatibility: Oxidizers
Shelf life: Indefinite if stored properly.
Section 11:
Toxicology Information
Acute Symptoms/Signs of exposure: Eyes: Stinging pain, watering of eyes, inflammation of eyelids and conjunctivitis. Skin:
Insensitivity to pain, feel of coolness or cold, skin looks white and feels hard and cold. Ingestion: Breath has sweet, organic
odor, mental confusion, drowsiness, nausea, vomiting and headache. Inhalation: Rapid irregular breathing, headache, fatigue,
mental confusion, nausea and vomiting, giddiness and poor judgment, convulsions and death.
Chronic Effects: Repeated/prolonged skin contact may cause dryness or rashes.
Sensitization: none expected
Guaiacol: LD50 [oral, rat]; 520 mg/kg; LC50 [rat]; N/A; LD50 Dermal [rabbit]; 5020 mg/24H Severe
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
EC50 – 25.9mg/l – 48h – Daphnia magna.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Listed (201-964-7).
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not reulated by TDG.
Regulatory Information
WHMIS Canada: Not WHMIS controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Other Information
Current Issue Date: December 21, 2011
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
12/21/2011
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
Page 46
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
Hydrogen Peroxide, 3%
MSDS # 345.00
Section 1:
Page 1 of 2
Product and Company Identification
Hydrogen Peroxide, 3%
Synonyms/General Names: N/A
Product Use: For educational use only
Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Hazards Identification
HMIS (0 to4)
Health
1
Fire Hazard
0
Reactivity 1
Clear, colorless liquid, slight odor
WARNING! Oxidizing agent and body tissue irritant.
Target organs: None known.
This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).
Section 3:
Composition / Information on Ingredients
Hydrogen Peroxide, 35% (7722-84-1), 3%.
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Water (7732-18-5), 97%.
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Do not induce vomiting. Rinse mouth with cold water. Give victim 1-2 cups of
water or milk to drink.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Noncombustible solution. When heated to decomposition, produces oxygen gas.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
1
0
1
Accidental Release Measures
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a
sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.
Section 7:
Handling and Storage
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Use NIOSH-approved respirator with an acid/organic
cartridge. Exposure guidelines Hydrogen Peroxide: OSHA PEL: 1.4 mg/m3; ACGIH TLV: 1.4 mg/m3; STEL:N/A.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
12/21/2011
250-7462 v.5/12
Page 47
Enzyme Activity: Teacher’s Guide
Kit # 36-7413
Material safety data sheets
Material Safety Data Sheet
MSDS # 345.00
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Hydrogen Peroxide, 3%
Scholar Chemistry
Physical and Chemical Properties
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
H2O2.
34.01.
1.01 g/mL @ 20°C.
0.7.
0°C.
100°C.
14.
N/A.
N/A.
Section 10:
Clear, colorless liquid.
Slight odor.
N/A.
Completely soluble in water.
<1
(Butyl acetate = 1).
N/A.
(log POW).
N/A.
N/A.
N/A.
Stability and Reactivity
Avoid heat and ignition sources.
Stability: Instable, many materials will catalyze the decomposition of hydrogen peroxide to produce oxygen, water, and heat.
Incompatibility: Reducing agents, alkalis, organic materials, metals, acids, bases, metal salts, dust and dirt contaminants and
flammable substances.
Shelf life: Fair shelf life, store in a cool, dry environment.
Section 11:
Toxicology Information
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath.
Chronic Effects: Repeated/prolonged skin contact may cause thickening, blackening or cracking. Repeated eye exposure may
cause corneal erosion or loss of vision.
Sensitization: none expected
Hydrogen Peroxide: LD50 [oral, rat]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial): Toxic to beneficial microorganisms (e.g. soil and sewage treatment microorganisms).
Do not release to environment.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Listed (231-765-0) .
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Not regulated by TDG .
Regulatory Information
WHMIS Canada: Not WHMIS controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Other Information
Current Issue Date: December 21, 2011
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all inclusive. The information
relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any particular process. Because safety standards and
regulations are subject to change and because S&C has no continuing control over the material, those handling, storing or using the material should satisfy themselves that they have
current information regarding the particular way the material is handled, stored or used and that the same is done in accordance with federal, state and local law. S&C makes no
warranty, expressed or implied, including (without limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with
respect to fitness for any particular use.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
12/21/2011
US: www.wardsci.com
Canada: www.wardsci.ca
250-7462 v.5/12
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