AP® INVESTIGATION #5 - Wikispaceshinton.harvardmedk12.wikispaces.net/file/view/Ward's... · Objectives of the lab as described in AP Biology Investigative Labs: An Inquiry-Based

AP® INVESTIGATION #5 CELL PROCESSES: PHOTOSYNTHESIS ! TEACHER’S GUIDE Kit # 36W7405

©2012, Ward’s Natural Science

All Rights Reserved, Printed in the U.S.A.

US: www.wardsci.com

Canada: www.wardsci.ca

250-7455 v.7/12

ABSTRACT . . . . . . . . . . . . . . . . . . . . . . . . . . . 1GENERAL OVERVIEW . . . . . . . . . . . . . . . . . . . . . 2RECORDING DATA . . . . . . . . . . . . . . . . . . . . . . . 3MATERIALS CHECKLIST . . . . . . . . . . . . . . . . . . . . . 5CURRICULUM ALIGNMENT . . . . . . . . . . . . . . . . . . . 6LEARNING OBJECTIVES . . . . . . . . . . . . . . . . . . . . . 7TIME REQUIREMENTS . . . . . . . . . . . . . . . . . . . . . 7SAFETY PRECAUTIONS . . . . . . . . . . . . . . . . . . . . . 8PRE"LAB PREPARATION . . . . . . . . . . . . . . . . . . . . 9COPY OF STUDENT GUIDE CONTENTS #WITH TEACHER ANSWER KEY$

BACKGROUND . . . . . . . . . . . . . . . . . . . . . . . 10

PART 1: PLANT PIGMENTS AND CHROMATOGRAPHY #STRUCTURED INQUIRY$ . . . . . . . . . . . . . . . . . . 13PART 2: FLOATING DISC ASSAY #GUIDED INQUIRY$ . . . . . . . 15PART 3: DESIGN AN EXPERIMENT #OPEN INQUIRY$ . . . . . . 18MATERIAL SAFETY DATA SHEETS . . . . . . . . . . . . . . . 20

Table of Contents

**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.

AP® INVESTIGATION #5: CELL PROCESSES: PHOTOSYNTHESIS ! TEACHER’S GUIDE Kit # 36W7405



US: www.wardsci.com


250-7455 v.7/12

Page 1

ABSTRACTThis lab illustrates the ability of plants to capture, store, and use light energy for growth and

reproduction. Part 1 is a structured inquiry activity in which students visualize the pigments present

in various plant leaves through the use of basic paper chromatography techniques. Part 2 is a guided

inquiry� activity� in� which� the� students� indirectly� measure� the� rate� of� photosynthesis� under� specific� environmental� conditions� using� the� floating� disc� assay.� Students� also� observe� how� interactions� of� the� cells� with� the� environment� enable� both� uptake� of� specific� components� necessary� for� photosynthesis� and expulsion of waste products that result from cellular metabolism. Part 3 is an open inquiry

activity, in which students design an experiment that allows them to further explore the process of

photosynthesis.




US: www.wardsci.com


250-7455 v.7/12

Page 2

GENERAL OVERVIEWThe 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 experiment 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, s/he 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 experiment

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.




US: www.wardsci.com


250-7455 v.7/12

Page 3

RECORDING DATA IN A LABORATORY NOTEBOOK

All of the Ward’s Investigations assume that students will keep a laboratory notebook for student-

directed 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 experiment 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)




US: www.wardsci.com


250-7455 v.7/12

Page 4

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

experiment� 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� a� specific� concept� (XXXX)� account� for� the� described� findings� (YYYY)?

•� Describe� a� method� that� could� be� used� to� determine� a� given� concept/observation� (XXXX).




US: www.wardsci.com


250-7455 v.7/12

Page 5

MATERIALS INCLUDED IN KITUnits per kit Description

1 Aluminum foil, 12” x 25’ roll

20 Syringe, NON-sterile, 10 mL

1 Sodium bicarbonate, 50 g

15 Graduated pipets, 6’’

1 ScholAR Chemistry New MSDS CD

8 One-hole paper punch

4 Film, Rainbow: 8-1/2 x 11

� 1� Buffer� set:� � Includes� envelopes� of� pH� 2-11� (one� each,� for� a� total� of� 10� envelopes),

500 mL buffer

1 pkg. Chromatography paper strips

8 Glass vials with caps

1 pkg./4 Chromatography solvent, 30 mL

50 Clear plastic cups

1 Instructions (this booklet)

MATERIALS NEEDED BUT NOT PROVIDED OPTIONAL MATERIALS !NOT PROVIDED"Lab Notebook

Timers

Light source

Baby spinach leaves

Forceps

Rulers

Wax pencil

Coins (quarters or dimes)

Scissors

Fume hood

Liquid soap (dishwashing liquid)

MATERIALS CHECKLIST

For a list of replacement items, visit: www.wardsci.com, and click on the AP Biology tab for this kit/item #.

Light sensor ( item 9200003 or other)

Thermometer

Variety of collected plant leaves

Ring stand with ring

Light bulbs, variety (40 watt, 100 watt, 150 watt suggested )

Balance or scale

PAR (photosynthetic active radiation) meter




US: www.wardsci.com


250-7455 v.7/12

Page 6

CURRICULUM ALIGNMENTBig Ideas

Big Idea 2: Biological systems utilize energy and molecular building blocks to grow, to

reproduce, and to maintain homeostasis.

Also connects to:

Big Idea 1: The process of evolution drives the diversity and unity of life.

Big Idea 4: Biological systems interact, and these interactions possess complex properties

Enduring Understandings

1B1: Organisms share many conserved core processes and features that evolved and are widely

distributed among organisms today.

2A1: All living systems require constant input of free energy.

2A2: Organisms capture and store free energy for use in biological processes.

2B3: Eukaryotic cells maintain internal membranes that partition the cell into specialized regions

(e.g., chloroplasts).

4A2: The structure and function of subcellular components, and their interactions, provide

essential cellular processes.

4A6: Interactions among living systems and their environment result in the movement of matter

and energy.

Science Practices

1.4 The student can use representations and models to analyze situations or solve problems

qualitatively and quantitatively.

2.2 The student can apply mathematical routines to quantities that describe natural phenomena.

3.1� The� student� can� pose� scientific� questions.

6.1 The student can justify claims with evidence.

6.2 The student can construct explanations of phenomena based on evidence produced through

scientific� practices.

7.2 The student can connect concepts in and across domain(s) to generalize or extrapolate in and/

or across enduring understandings and/or big ideas.

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

Kit # 36W7405

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-Based Approach (2012). This is a publication of the College Board that can be found at http://advancesinap.collegeboard.org/science/biology/lab.




US: www.wardsci.com


250-7455 v.7/12

Page 7

LEARNING OBJECTIVESThe� student� is� able� to� describe� specific� examples� of� conserved� core� biological� processes� and� features shared by all domains or within one domain of life, and how these shared, conserved core

processes and features support the concept of common ancestry for all organisms (1B1 & SP 7.2.)

The� student� is� able� to� justify� the� scientific� claim� that� organisms� share� many� conserved� core� processes and features that evolved and are widely distributed among organisms today (1B1 & SP

6.1).

The� student� is� able� to� justify� the� scientific� claim� that� free� energy� is� required� for� living� systems� to� maintain organization, to grow, or to reproduce, but that multiple strategies exist in different living

systems (2A1 & SP 6.1).

The� student� is� able� to� use� representations� to� pose� scientific� questions� about� what� mechanisms� and� structural features allow organisms to capture, store, and use free energy (2A2 & SP 1.4, SP 3.1).

The student is able to use representations and models to describe differences in prokaryotic and

eukaryotic cells (2B3 & SP 1.4).

The� student� is� able� to� construct� explanations� based� on� scientific� evidence� as� to� how� interactions� of� subcellular structures provide essential functions (4A2 & SP 6.2).

The student is able to apply mathematical routines to quantities that describe interactions among

living systems and their environment, which result in the movement of matter and energy (4A6 &

SP 2.2).

TIME REQUIREMENTS

Part 1: Plant Pigments and Chromatography

(Structured Inquiry)

45 minutes.

Optional:

Start Part 2 during solvent migration.

Part 2: Floating Disc Assay

(Guided Inquiry)45 minutes

Part 3: Open InquiryVaries, depending on students’

experiment designs

‹

‹

‹

‹

‹

‹

‹




US: www.wardsci.com


250-7455 v.7/12

Page 8

SAFETY PRECAUTIONS Lab-Specific� Safety

Chromatography solution (used in paper chromatography) is an irritant to the skin and eyes, use

with precaution.

Chromatography� solvent� is� extremely� flammable,� a� serious� health� hazard,� and� moderately� reactive. Use this chemical in an approved fume hood.

General� Safety

The teacher should 1) be familiar with safety practices and regulations in his/her school (district

and state) and 2) 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. This is

a good way to identify students with allergies (e.g., latex) so that you (and they) will be reminded

of� specific� lab� materials� that� may� pose� 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.

Require students to remove all dangling jewelry and tie back long hair before they begin.

Remind students to read all instructions, Material Data Safety 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. The most updated version of most MSDSs can usually be found on the chemical

manufacturer’s website. (Note that in this particular lab, there are no materials that require live care sheets.)

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� the� end� of� the� lab:

All laboratory bench tops should be wiped down with a 10% bleach solution or disinfectant to

ensure cleanliness.

Remind students to wash their hands thoroughly with soap and water before leaving the

laboratory.

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹

‹




US: www.wardsci.com


250-7455 v.7/12

Page 9

PRE#LABORATORY PREPARATIONFor� all� parts� of� the� lab:

1. Make� copies� of� Student� Guide.

Copy all pages of the student copymaster prior to starting class.

For Part 1: Structured� Inquiry� –� Plant� Pigments� &� Chromatography

1.� Obtain� (and� cut,� if� necessary)� chromatography� paper� strips� to� fit� the chromatography vial.

2. Obtain fresh baby spinach leaves, and coins.

3. A fume hood or well ventilated area is necessary for working

with the chromatography solvent.

For� Part� 2:� Guided� Inquiry� –� Floating� Disc� Assay

1. Soak� the� baby� spinach� leaves.

Prepare the fresh spinach leaves by soaking them in water

overnight under a light source. This initiates the process

of� photosynthesis,� and� the� intercellular� spaces� fill� with� air� (increasing buoyancy).

2. Prepare� 0.2%� sodium� bicarbonate� solution�

Add 2 grams of sodium bicarbonate to 1 L of distilled water and

mix. (1 g into 100 mL = 1%)

3. Dilute� the� soap� solution.

To prepare the diluted soap solution, add 1.5 mL of dishwashing

liquid to 250 mL 0.2% sodium bicarbonate solution. Mix gently

and allow the solution to sit, in order to dissipate the soap suds.

NOTES




US: www.wardsci.com


250-7455 v.7/12

Page 10

Growth and reproduction are heavily energy-dependent processes that

have driven organisms to evolve strategies, structures, and processes

that enable them to capture, utilize, and store free energy. Free

energy is available in the environment in a multitude of forms, and

autotrophs and heterotrophs employ different approaches to harvest the

energy they need to live. Photosynthesis and chemosynthesis enable

autotrophs (or primary producers) to obtain free energy directly from

their surroundings, whereas heterotrophs employ cellular respiration to

produce energy. They must seek sources of food, and utilize the energy

stored in carbon compounds produced by other organisms.

Autotrophs are “self-feeders”. This name is derived from the Greek

words auto (meaning “self”) and trophos (meaning “feeder”).

Multicellular plants are examples of photoautotrophs, organisms that

produce organic molecules from light energy. Photosynthesis is the

name of the process whereby photoautotrophs capture light energy

present in the environment for use in growth, reproduction, and

maintaining homeostasis.

The set of chemical reactions involved in photosynthesis transforms

the substrates carbon dioxide and water, into glucose (a simple

carbohydrate) and oxygen. The chemical bonds of the glucose molecule

serve to store the transformed light energy until it is harvested in the

process of respiration. Measuring the oxygen produced in this reaction

is one way to measure the rate of photosynthesis.

In plants, chloroplasts are required to capture the energy that drives

the reaction. Chloroplasts are membrane-bound organelles that

contain a variety of photoreactive pigments, including the primary

photosynthetic pigments - chlorophylls. Chlorophyll molecules absorb

light� energy� in� the� red� and� blue� portions� of� the� spectrum,� and� reflect� green wavelengths, making the chlorophyll, and thus plants, appear

green to us. Accessory pigments in chloroplasts and leaves have

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.

‹

‹

‹


BACKGROUND




US: www.wardsci.com


250-7455 v.7/12

Page 11

BACKGROUND !CONTINUED"

other roles in regulating light energy. For example, yellow/orange/red

carotenoids absorb high frequency ultraviolet light that can damage

DNA. The pigments in chloroplasts are embedded in stacks of

membrane and are hydrophobic. Some of the common pigments found

in leaves are listed in order from most polar (hydrophilic) to least polar

(hydrophobic) as follows: chlorophyll b, chlorophyll a, phaeophytin b,

phaeophytin a, xanthophylls, carotene.

The terrestrial plant cells that are specialized for photosynthesis (and

thus contain most of the chloroplasts and chlorophyll) are in the leaves.

The structure of the leaf has evolved to regulate the photosynthetic

reactions by regulating how the substrates of carbon dioxide and water

are brought together with light to form carbohydrates and release

oxygen. Guard cells in the lower epidermis of the leaf form pores

called stomata that can be opened or closed to regulate gas exchange in

the leaf under different environmental situations. One of the substrates

for the photosynthetic reaction, CO2, enters the leaf through the open

stomata.� The� other� substrate,� H2O, enters the leaf mostly through the

vascular system of the plant, the xylem tubules. The palisade cell

layer of a leaf has access to both substrates and consists of cylindrical

cells that contain large numbers of chloroplasts. These are the cells

primarily responsible for photosynthesis and thus energy capture in the

plant. Carbohydrates (products of photosynthesis) can be transported

out to other parts of the plant through the phloem of the vascular

system. Oxygen (another product of photosynthesis) passes into the

spongy mesophyll layer. This layer contains air chambers that expand

as the oxygen is produced and then released through the stomata into

the environment. Alternatively, the carbohydrates and oxygen can be

used as substrates for respiration in the leaf cells.

NOTES




US: www.wardsci.com


250-7455 v.7/12

Page 12

SAFETY PRECAUTIONS When� working� with� the� chromatography� solvent,� use� a�

chemical� fume� hood� or� proper� ventilation.�

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 for the material is the vendor. 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 10%

bleach solution or disinfectant to ensure cleanliness.

Wash your hands thoroughly with soap and water before leaving

the laboratory.

‹

‹

‹

‹

‹

NOTES




US: www.wardsci.com


250-7455 v.7/12

Page 13

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.

‹

‹

PART 1 $ STRUCTURED INQUIRY: PLANT PIGMENTS AND CHROMATOGRAPHY

MATERIALS NEEDED PER LAB GROUP1 Chromatography vial, with cap (provided)

1 Wax pencil

1 Disposable transfer pipet (provided)

1 mL Chromatography solvent

(acetone:ethyl alcohol::1:1. (provided)

1 Chromatography paper strip (provided)

1 Sharp pencil

1 Ruler (metric)

1 Pair of scissors

1 Piece of fresh (pre-soaked) spinach

1 Coin (a quarter works well)

1 Pair of forceps

PART 1 $ PROCEDURE: STRUCTURED INQUIRY1. Obtain a chromatography paper strip.

NOTE: Be sure to handle the chromatography paper by

the edges. Do not touch the surface of the strip with your

fingertips� as� the� oils� from� your� fingers� will� interfere� with� the� chromatogram.

2. Measure 1.5 cm from one end of the chromatography strip and

lightly draw a pencil line across the width of the strip. This is

the point of application (see Step 4).

3. Use a pair of scissors to cut off two small pieces below the

pencil line to form a pointed end (see Figure 1). The pointed end

will be referred to as the bottom end of the chromatogram.

4. Obtain a well-hydrated leaf of spinach which, has been pre-

soaked to jump-start the process of photosynthesis. Place it

over the point of application on the chromatography strip. Rub

or roll the ribbed edge of a coin over the spinach leaf to extract

the pigments. Repeat 5-10 times with different portions of the

spinach leaf, making sure you are rubbing the coin over the

pencil line (see Figure 2).

qqqq

qqqqqqq

‹


Figure 1

Figure 2




US: www.wardsci.com


250-7455 v.7/12

Page 14

5. Obtain a chromatography vial from your teacher and label it

with your initials using a permanent marker or wax pencil.

CAUTION:� Steps� 6-11� should� be� performed� under� a�

fume� hood� or� in� a� well� ventilated� area.

6. Go to a fume hood or a well ventilated area and remove the cap

from the chromatography vial. Using a disposable pipet, add

1 mL of chromatography solvent to the vial. The solvent is a

volatile organic compound (hydrophobic) and a fume hood is

required to capture the volatile fumes.

7. Carefully place the chromatography paper strip into the vial so

that the pointed end is barely immersed in the solvent. Do not

immerse the pigments into the solvent.

8. Cap the vial and leave it undisturbed. Observe as the solvent is

drawn up the chromatography paper strip by capillary action.

9. Record the different colors you observe as they separate along

the strip.

10. When the solvent reaches a level approximately 1 cm from the

top of the strip, remove the cap from the vial. Using forceps,

remove the strip from the vial. This is your chromatogram.

11. The solvent will evaporate quickly; immediately use a pencil

to mark the location of the solvent at the top end of the

chromatography paper strip. This is the solvent front.

NOTE:� You� will� need� this� location� to� identify� the� distance� the chromatography solvent traveled.

12. List and record the pigment colors or names. Once the strip has

completely dried, mark the middle of each pigment band on the

chromatography paper strip with a pencil.

13. Using a metric ruler, measure the distance from the original

pencil line with the spinach extract to the solvent front and each

mark you made on the pigment bands (see Figure 3). Record

these distances in millimeters (mm).

14. Calculate the Rf value for each pigment on your chromatogram.

Rf =

Distance traveled by component from point of application

Distance traveled by solvent from point of application

‹

‹

PROCEDURE $ PART 1: STRUCTURED INQUIRY !CONTINUED"

NOTES

Figure 3




US: www.wardsci.com


250-7455 v.7/12

Page 15

PART 2 $ GUIDED INQUIRY: FLOATING DISC ASSAY

MATERIALS NEEDED PER LAB GROUP10 mL 0.2% sodium bicarbonate solution with dish soap added

100 mL 0.2% sodium bicarbonate solution

1 Disposable beaker 100 mL (or plastic cup)

1 One-hole punch

1 Timer

1 Light source

1 10 mL syringe

1 Piece of fresh (pre-soaked) spinach

PART 2 $ PROCEDURE: GUIDED INQUIRY1. Obtain a leaf of spinach that is well hydrated, and has been pre-

soaked to jump-start the process of photosynthesis.

2. Use a one-hole punch to cut discs from the leaf (at least 10 discs

per trial).

Each trial requires at least 10 leaf discs. When cutting discs,

avoid major veins, aberrant tissue, and leaf edges. Make an

effort to obtain consistent discs.

3. Remove the plunger from the barrel of a 10 mL syringe. Place

the leaf discs inside the barrel, and tap to get them down to the

bottom of the syringe barrel. Replace the plunger at the top of

the syringe barrel and depress it. Push the plunger in enough to

expel most of the air from the syringe.

4. Fill the barrel of the plunger half full with the sodium-

bicarbonate/dish soap mixture. Invert the syringe (air bubble

floats� to� syringe� tip)� and� depress� the� plunger� farther� to� remove� air from the syringe.

NOTE:� The liquid soap in the sodium bicarbonate aids in

wetting the normally hydrophobic surface of the leaf. The

surface tension of the water is broken by the addition of the

soap, and the leaf surface can become wet with the solution

in� order� to� allow� it� to� infiltrate� the� pores� on� the� leaf� surface,� and� fill� the� intercellular� spaces� in� the� spongy� mesophyll.

qqqqqqqq

‹

‹

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.

‹

‹





US: www.wardsci.com


250-7455 v.7/12

Page 16

5.� � Hold� your� finger� over� the� hole� at� the� end� of� the� syringe� and� draw back on the plunger to form a vacuum within the chamber

of� the� syringe.� Hold� this� for� 10� seconds� while� swirling� the� syringe to wet the leaf discs. Repeat until all of the leaf discs

sink under normal pressure. This means that the solution has

infiltrated� the� spongy� layer� of� the� leaf.�

Do not use leaf discs that do not sink.

Additional soap (up to double the concentration) may be

added if leaves will not sink.

A negative control may be set up with water and soap, but

no bicarbonate.

6. Remove the plunger from the syringe and pour the discs and

solution into a clear plastic cup containing 0.2% sodium

bicarbonate (or plain water for a negative control) at a depth

of about 3 cm. Be sure to place all of the discs in the bottom of

the cup. The sodium bicarbonate serves as the source of CO2

necessary for photosynthesis to occur. Keep the depth of the

solution in the cup consistent throughout the trials.

7. Place the reaction vessels under a bright light source. Start the

timer immediately.

Hint:� � Put� the� light� source� as� close� to� the� experimental� beakers as possible.

8.� Record� the� number� of� discs� that� are� floating� at� 30� second� intervals.

9. Graph your results over time for bicarbonate and water-only

conditions.

10. Explain your results.

‹

‹

‹

‹

PROCEDURE $ PART 2: GUIDED INQUIRY !CONTINUED"

NOTES




US: www.wardsci.com


250-7455 v.7/12

Page 17

ASSESSMENT QUESTIONS !FOR PARTS 1 AND 2"

1. Which pigment migrated the farthest on the chromatogram? Explain how this migration occurred.

Answers should indicate that the same number and color pigments migrated in the same order up the chromatography strip. Explanations should include references to the relative solubility of the pigments (based on their relative polarities) in the chromatography solvent. Students must use logical reasoning and data from their experiment as evidence to support their conclusions.

2. What does the Rf value represent? If you were to perform your experiment on a chromatography

paper twice the length of the one used, would your Rf values still be the same?

The Rf value indicates the distance the pigment moved (from the point of sample application) relative

to the distance the solvent moved (from the point of sample application). The length of the paper does not affect the R

f values since it is proportional to the distance moved.

3.� How� do� plant� pigments� and� the� absorption� spectrum� relate� to� photosynthesis?

Students should discuss the roles of various pigments in photosynthesis, the pigments present in various plants, and how light of different wavelengths is available to/utilized by plants with different pigments to optimize photosynthesis for that organism.

4. Name at least four parameters that will affect the rate of photosynthesis as measured by this

investigation.� How� does� each� parameter� have� bearing� on� the� reactions� of� photosynthesis?�

Some parameters are: temperature, pH, light intensity, wavelength, availability of CO2, availability

of water. There will be an optimal or peak range for each of these parameters, with the rate of photosynthesis� tapering� off� as� the� optimal� conditions� are� not� satisfied.

5. What might happen if you were to remove all light from the setup after the discs have all become

buoyant? Describe what you would see. Explain why this would occur with relation to cellular

processes like respiration.

If all light were removed after the discs become buoyant then the rate of photosynthesis would drop off sharply, and the competing process of cellular respiration would take over, utilizing the available O

2 and glucose in the leaf discs (that were produced by photosynthesis), producing CO

2,

and eventually causing the discs to sink. Cellular respiration occurs in both the light and dark. Photosynthesis is the dominant reaction in the presence of light, but cannot occur without light, so cellular respiration takes over in the dark. What is measured with O

2 production in the light is the

net rate of photosynthesis and respiration.




US: www.wardsci.com


250-7455 v.7/12

Page 18

PART 3 $ OPEN INQUIRY: DESIGN AN EXPERIMENTWhat questions occurred to you as you performed the investigations?

Now that you are familiar with photosynthetic pigments,

chromatography, and a photosynthesis rate assay, design an experiment

to investigate one of your questions.

Questions may include:

How� is� the� rate� of� photosynthesis� in� leaves� related� to� pigments� in� the� leaf?� How� does� the� amount� (intensity)� or� wavelength� of� light� affect� the� rate� of� photosynthesis?� Does� temperature� or� pH� affect� the� rate of photosynthesis? Is there a variation in chloroplast density in

different leaves that affects photosynthesis? Does the age of the leaf

or plant that the leaf came from affect the rate of photosynthesis?

Does the habitat in which the plant evolved affect pigments or rates of

photosynthesis? Does the time of year that the leaf was collected affect

pigments or photosynthetic rate? What other aspects of the leaf or

plant environment might affect photosynthetic rates?

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 experiment 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 weights and volumes used.

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 ques-

tions, 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� a� specific� concept� (XXXX)� account� for� described� findings� (YYYY)?

Describe a method that could

be used to determine a given

concept/observation� (XXXX).

‹

‹

‹

‹

‹





US: www.wardsci.com


250-7455 v.7/12

Page 19

8. The results should be recorded (including drawings, photos, data

print-outs).

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 experiment design, statistical

significance� and� uncontrolled� variables.

12. Further study direction should be considered.

NOTES

PART 3: OPEN INQUIRY !CONTINUED"

MATERIAL SAFETY DATA SHEET

US: www.wardsci.com Canada: www.wardsci.ca

250-7455 v.7/12Page 20

MSDS Reprinted for Customer Convenience



Material Safety Data Sheet Page 1 of 2

© 2009, Scholar Chemistry. All Rights Reserved. 1/20/2012

Buffer Capsule, pH 2.00 MSDS # 126.30

Section 1: Product and Company Identification Buffer Capsule, pH 2.00

Synonyms/General Names: pH 2 Buffer Capsule Product Use: For educational use onlyManufacturer: 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: Hazards Identification Capsule containing white powder; no odor. HMIS (0 to 4)

CAUTION! Powder is a body tissue irritant. Target organs: None known

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 Sulphamic Acid (5329-14-6) 30 to 40% Potassium Biphthalate ( 877-24-7) 60 to 70%

Section 4: First Aid Measures Always seek professional medical attention after first aid measures are provided.

Eyes: Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.Skin: Immediately flush skin with excess water for 15 minutes while removing contaminated clothing. Ingestion: Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.

Induce vomiting immediately. Inhalation: Remove to fresh air. If not breathing, give artificial respiration.

Section 5: Fire Fighting Measures Noncombustible solution. When heated to decomposition, emits acrid fumes. 0 Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire. 0 0

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 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.

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 ProtectionUse 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.

Health 1 Fire Hazard 0 Reactivity 0

AP® INVESTIGATION #5: CELL PROCESSES: PHOTOSYNTHESIS ! TEACHER’S GUIDE


Kit # 36W7405


250-7455 v.7/12Page 21




MSDS # 126.30 Buffer Capsule, pH 2.00 Scholar Chemistry Section 9: Physical and Chemical Properties Molecular formula N/A. Appearance Capsule with white solid.Molecular weight N/A. Odor No odor.Specific Gravity N/A Odor Threshold N/A. Vapor Density (air=1) N/A Solubility Complete. Melting Point N/A Evaporation rate N/A (Butyl acetate = 1). Boiling Point/Range N/A. Partition Coefficient N/A (log POW). Vapor Pressure (20°C) N/A. pH 2.0, acidic. Flash Point: N/A. LEL N/A. Autoignition Temp.: N/A. UEL N/A.

N/A = Not available or applicable

Section 10: Stability and Reactivity Avoid heat and moisture.Stability: Stable under normal conditions of use and storage. Incompatibility: Acids, alkalis, Shelf life: Indefinite if stored properly.

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: 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: Transport Information DOT Shipping Name: Not regulated by DOT. Canada TDG: Not regulated by TDG. DOT Hazard Class: Hazard Class: Identification Number: UN Number:

Section 15: Regulatory Information EINECS: Not listed . WHMIS Canada: Not WHMIS Controlled. TSCA: All components are listed or are exempt. 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: January 20, 2012 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.



250-7455 v.7/12Page 22












CAUTION! Powder is a body tissue irritant. Target organs: None known


Section 3: Composition / Information on Ingredients Sulphamic Acid (5329-14-6) 20 to 30% Potassium Biphthalate ( 877-24-7) 70 to 80%










Section 8: Exposure Controls / Personal ProtectionUse 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.




Kit # 36W7405


250-7455 v.7/12Page 23







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: 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.









250-7455 v.7/12Page 24











CAUTION! Powder may be a slight body tissue irritant. Target organs: None known


Section 3: Composition / Information on Ingredients Potassium Biphthalate ( 877-24-7) 100%










Section 8: Exposure Controls / Personal ProtectionUse 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.





Kit # 36W7405


250-7455 v.7/12Page 25







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: 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.









250-7455 v.7/12Page 26













Section 3: Composition / Information on Ingredients Potassium Biphthalate ( 877-24-7), 75 to 85% Sodium Phosphate Dibasic, (7558-79-4), 15 to 25%










Section 8: Exposure Controls / Personal ProtectionUse 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.





Kit # 36W7405


250-7455 v.7/12Page 27







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: 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.









250-7455 v.7/12Page 28













Section 3: Composition / Information on Ingredients Sodium Phosphate Dibasic, (7558-79-4), 15 to 25% Potassium Phosphate monobasic (7778-77-0), 75 to 85%










Section 8: Exposure Controls / Personal ProtectionUse 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 6 Buffer: OSHA PEL: N/A, ACGIH: TLV: N/A, STEL: N/A.




Kit # 36W7405


250-7455 v.7/12Page 29







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: 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.









250-7455 v.7/12Page 30




























Kit # 36W7405


250-7455 v.7/12Page 31




MSDS # 132.30 Buffer Capsule, pH 7.00 Scholar Chemistry Section 9: Physical and Chemical Properties Molecular formula N/A. Appearance Capsule with white solid.Molecular weight N/A. Odor No odor.Specific Gravity N/A Odor Threshold N/A. Vapor Density (air=1) N/A Solubility Complete. Melting Point N/A Evaporation rate N/A (Butyl acetate = 1). Boiling Point/Range N/A. Partition Coefficient N/A (log POW). Vapor Pressure (20°C) N/A. pH 7.0, neutral. Flash Point: N/A. LEL N/A. Autoignition Temp.: N/A. UEL N/A.












250-7455 v.7/12Page 32




























Kit # 36W7405


250-7455 v.7/12Page 33
















250-7455 v.7/12Page 34













Section 3: Composition / Information on Ingredients Sodium Carbonate, (497-19-8), 20 to 30% Sodium Bicarbonate (144-55-8), 70 to 80%















Kit # 36W7405


250-7455 v.7/12Page 35
















250-7455 v.7/12Page 36













Section 3: Composition / Information on Ingredients Sodium Carbonate, (497-19-8), 50 to 70% Sodium Bicarbonate (144-55-8), 30 to 50%














Kit # 36W7405


250-7455 v.7/12Page 37

















250-7455 v.7/12Page 38













Section 3: Composition / Information on Ingredients Sodium Phosphate, tribasic, (7601-54-9), 20 to 30% Sodium Bicarbonate (144-55-8), 70 to 80%














Kit # 36W7405


250-7455 v.7/12Page 39

















250-7455 v.7/12Page 40






Chromatography Solvent (9:1) MSDS # 185.00 Section 1: Product and Company Identification

Chromatography Solvent (9:1) Synonyms/General Names: Petroleum Ether/Acetone (9:1) Product Use: For educational use only Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.



Section 2: Hazards Identification Clear, colorless liquid, mild sweet odor. HMIS (0 to 4) DANGER! Flammable liquid and moderately toxic by ingestion. Flammable liquid, keep away from all ignition sources. Target organs: Eyes, skin, mucous membranes, respiratory and central nervous system. This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).

Section 3: Composition / Information on Ingredients Petroleum Ether, (8032-32-4), 90% . Acetone, (67-64-1), 10%.


Eyes: Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally. Skin: Immediately flush skin with excess water for 15 minutes while removing contaminated clothing. Ingestion: Call Poison Control immediately. Aspiration hazard. Rinse mouth with cold water. Give victim 1-2 tbsp of

activated charcoal mixed with 8 oz water. Inhalation: Remove to fresh air. If not breathing, give artificial respiration.

Section 5: Fire Fighting Measures Class 1B Flammable Liquid. When heated to decomposition, emits acrid fumes 4 Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire. 1 0 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: 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 Red 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: Petroleum Ether: OSHA PEL: 2000 mg/m3, ACGIH TLV: N/A, STEL: N/A, Acetone: OSHA PEL: 2400 mg/m3, ACGIH TLV: 1188 mg/m3, STEL: 1782 mg/m3.




Kit # 36W7405


250-7455 v.7/12Page 41




MSDS # 185.00 Chromatography Solvent (9:1) Scholar Chemistry Section 9: Physical and Chemical Properties Molecular formula Mixture. Appearance Clear, colorless liquid. Molecular weight N/A. Odor Mild sweet odor . Specific Gravity 0.67 g/mL @ 20°C. Odor Threshold N/A. Vapor Density (air=1) 2.5 (Pet. ether). Solubility Insoluble. Melting Point N/A. Evaporation rate N/A (Butyl acetate = 1). Boiling Point/Range N/A. Partition Coefficient N/A (log POW). Vapor Pressure (20°C) N/A. pH N/A. Flash Point: -45.6°C (-50°F). LEL 1.1%. Autoignition Temp.: 288°C (550.4°F). UEL 5.9 %.


Section 10: Stability and Reactivity Avoid heat and ignition sources. Stability: Stable under normal conditions of use. Incompatibility: Oxidizing materials, plastic, rubber. 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 Petroleum Ether: LD50 [oral, rat]; 7060 mg/kg; LC50 [rat]; 20,000 mg/l (10 hours); LD50 Dermal [rabbit]; 20 mg/24H MOD Acetone: LD50 [oral, rat]; 5800 mg/kg; LC50 [rat]; 50,100 mg/l (8 hours); LD50 Dermal [rabbit]; 500 mg/24H MOD 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. Use a licensed chemical waste disposal firm for proper disposal.

Section 14: Transport Information DOT Shipping Name: Flammable liquid, n.o.s. Canada TDG: Flammable liquid, n.o.s. (Petroleum Ether and Acetone). (Petroleum Ether and Acetone). DOT Hazard Class: 3, pg II. Hazard Class: 3, pg II . Identification Number: UN1993. UN Number: UN1993.

Section 15: Regulatory Information EINECS: Not listed . WHMIS Canada: Not WHMIS controlled. TSCA: All components are listed or are exempt. 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.




250-7455 v.7/12Page 42



Prod

uct

MAT

ERIA

L SA

FETY

DAT

A SH

EET

CH

EMTR

EC24

Hou

r Em

erge

ncy

Pho

ne N

umbe

r (80

0) 4

24-9

300

The

info

rmat

ion

cont

aine

d he

rein

is fu

rnis

hed

with

out w

arra

nty

of a

ny k

ind.

Em

ploy

ers

shou

ld u

se th

is in

form

atio

n on

ly a

s a

supp

lem

ent

to o

ther

info

rmat

ion

gath

ered

by

them

and

mus

t mak

e in

depe

nden

t det

erm

inat

ions

of s

uita

bilit

y an

d co

mpl

eten

ess

of in

form

atio

n fro

m a

ll so

urce

s to

ass

ure

prop

er u

se o

f the

se m

ater

ials

and

the

safe

ty a

nd h

ealth

of e

mpl

oyee

s. *

Haz

ardo

us M

ater

ials

Indu

stria

l Sta

ndar

ds.

Sect

ion

1C

hem

ical

Pro

duct

and

Com

pany

Info

rmat

ion

Sect

ion

5 Fi

re F

ight

ing

Mea

sure

s

Sect

ion

6 A

ccid

enta

l Rel

ease

Mea

sure

s

Sect

ion

7 H

andl

ing

& S

tora

ge

Sect

ion

8 Ex

posu

re C

ontr

ols

/ Per

sona

l Pro

tect

ion

Sect

ion

12

Ecol

ogic

al In

form

atio

n

Sect

ion

13

Dis

posa

l Con

side

ratio

ns

Sect

ion

14

Tran

spor

t In

form

atio

n

Sect

ion

16

Add

ition

al In

form

atio

n

NFP

A0

=M

inim

al1

=S

light

2=

Mod

erat

e

3=

Ser

ious

4=

Sev

ere

Syno

nym

s

Sect

ion

15

Reg

ulat

ory

Info

rmat

ion

Sect

ion

9 Ph

ysic

al &

Che

mic

al P

rope

rtie

s

Sect

ion

10

Stab

ility

& R

eact

ivity

Sect

ion

11

Toxi

colo

gica

l Inf

orm

atio

nSe

ctio

n 4

Firs

t Aid

Mea

sure

s

Che

mic

al N

ame

CA

S #

%

TLV

Uni

ts (A

CG

IH 2

001)

Sect

ion

3C

ompo

sitio

n / I

nfor

mat

ion

on In

gred

ient

s

Sect

ion

2 H

a]ar

ds Id

enti¿

catio

nE

mer

genc

y O

verv

iew

HM

IS *

0=

Min

imal

1=

Slig

ht2

=M

oder

ate

3

=S

erio

us4

=S

ever

e

Hea

lthFi

reR

eact

ivity

Con

tact

SS

0270

Janu

ary

12, 2

012

Jam

es A

. Ber

tsch

GEN

ERA

L ST

OR

AG

E C

OD

E G

REE

N

SOD

IUM

BIC

AR

BO

NAT

E, A

NH

YDR

OU

S

Sod

ium

bic

arbo

nate

, anh

ydro

us14

4-55

-810

0%N

one

esta

blis

hed.

CA

UTI

ON

!M

AY C

AU

SE

IRR

ITAT

ION

TO

SK

IN A

ND

EY

ES

.Av

oid

cont

act w

ith s

kin

and

eyes

. W

ash

thor

ough

ly a

fter h

andl

ing.

Sto

re in

a c

ool,

dry

plac

e aw

ay fr

om a

cids

and

aci

d fu

mes

. Ta

rget

org

ans:

Non

e kn

own.

Gen

eral

info

rmat

ion:

In

¿re

con

ditio

ns, w

ear a

NIO

SH�M

SH

A-a

ppro

ved

self-

cont

aine

d br

eath

ing

appa

ratu

s an

d fu

ll pr

otec

tive

gear

. Th

is m

ater

ial i

s co

mm

only

use

d to

e[t

ingu

ish ¿r

es.

Use

pro

per p

erso

nal p

rote

ctiv

e eq

uipm

ent a

s in

dica

ted

in S

ectio

n 8.

Rec

over

for u

se if

not

con

tam

inat

ed.

Sw

eep

or v

acuu

m u

p an

d pl

ace

in a

sui

tabl

e co

ntai

ner f

or p

rope

r dis

posa

l. W

ash

spill

are

a w

ith s

oap

and

wat

er.

Avoi

d ru

noff

into

sto

rm s

ewer

s an

d di

tche

s w

hich

lead

to w

ater

way

s.

Engi

neer

ing

cont

rols

: Fac

ilitie

s st

orin

g or

util

izin

g th

is m

ater

ial s

houl

d be

equ

ippe

d w

ith a

n ey

ewas

h fa

cilit

y an

d a

safe

ty s

how

er a

nd ¿

re e[t

ingu

ishi

ng m

ater

ial.

Per

sonn

el s

houl

d w

ear s

afet

y gl

asse

s, g

oggl

es, o

r fac

eshi

eld,

lab

coat

or a

pron

, app

ropr

iate

pro

tect

ive

glov

es.

Use

ade

quat

e ve

ntila

tion

to k

eep

airb

orne

con

cent

ratio

ns lo

w.

Res

pira

tory

pro

tect

ion:

Use

a c

hem

ical

fum

e ho

od a

nd�o

r wea

r a N

IOS

H�M

SH

A-a

ppro

ved

resp

irato

r.

Effe

cts

of o

vere

xpos

ure:

Thi

s pr

oduc

t is

a m

ild ir

ritan

t to

eyes

and

ski

n. S

ympt

oms

incl

ude

irrita

tion

or re

dnes

s of

eye

s or

ski

n. E

ye o

r ski

n di

seas

e an

d br

eath

ing

or re

spira

tory

dis

orde

rs m

ay b

e ag

grav

ated

by

expo

sure

to

dust

s pr

oduc

ed b

y th

is c

hem

ical

. M

ay c

ause

gas

troin

test

inal

dis

turb

ance

if in

gest

ed.

OR

L-R

AT L

D50

: 42

20 m

g�kg

IHL-

RAT

LC

50:

N�A

SK

N-R

BT

LD50

: N�A

Dat

a no

t yet

ava

ilabl

e.

Thes

e di

spos

al g

uide

lines

are

inte

nded

for t

he d

ispo

sal o

f cat

alog

-siz

e qu

antit

ies

only.

Fed

eral

regu

latio

ns m

ay a

p-pl

y to

em

pty

cont

aine

r. S

tate

and�o

r loc

al re

gula

tions

may

be

diffe

rent

. D

ispo

se o

f in

acco

rdan

ce w

ith a

ll lo

cal,

stat

e an

d fe

dera

l reg

ulat

ions

or c

ontra

ct w

ith a

lice

nsed

che

mic

al d

ispo

sal a

genc

y.

UN

/NA

num

ber:

N�A

Ship

ping

nam

e:

Not

Reg

ulat

ed.

Haz

ard

clas

s:

N�A

Pack

ing

grou

p:

N�A

Exce

ptio

ns:

N�A

ING

ESTI

ON

: Cal

l phy

sici

an o

r Poi

son

Con

trol C

ente

r im

med

iate

ly. I

nduc

e vo

miti

ng o

nly

if ad

vise

d by

app

ropr

iate

m

edic

al p

erso

nnel

.N

ever

giv

e an

ythi

ng b

y m

outh

to a

n un

cons

ciou

s pe

rson

.

INH

ALA

TIO

N: R

emov

e to

fres

h ai

r. If

not

bre

athi

ng, g

ive

arti¿

cial

resp

iratio

n. I

f bre

athi

ng is

dif¿

cult,

giv

e ox

ygen

. G

et m

edic

al a

ttent

ion.

EYE

CO

NTA

CT:

Che

ck fo

r and

rem

ove

cont

act l

ense

s. F

lush

thor

ough

ly w

ith w

ater

for a

t lea

st 1

5 m

inut

es, l

iftin

g up

per a

nd lo

wer

eye

lids

occa

sion

ally.

Get

imm

edia

te m

edic

al a

ttent

ion.

SKIN

CO

NTA

CT:

Rem

ove

cont

amin

ated

clo

thin

g. F

lush

thor

ough

ly w

ith m

ild s

oap

and

wat

er.

If irr

itatio

n oc

curs

, ge

t med

ical

atte

ntio

n.

Extin

guis

hing

Med

ia:

Use

any

med

ia s

uita

ble

for e

xtin

guis

hing

sup

porti

ng ¿

re.

Flas

h Po

int:

N�A

Aut

oign

ition

tem

pera

ture

: N�A

Expl

osio

n Li

mits

: Lo

wer

: N

�A

Upp

er:

N�A

MS

DS

No.

:R

evis

ion

Dat

e:A

ppro

ved

by:

Bak

ing

Sod

a; S

odiu

m H

ydro

gen

Car

bona

te

TSC

A-li

sted

, EIN

EC

S-li

sted

(205

-633

-8)

0 0 1 1

Phys

ical

sta

te:

Sol

id.

App

eara

nce:

Whi

te, c

ryst

allli

ne p

owde

r.O

dor:

No

odor

.pH

: N�A

Vapo

r pre

ssur

e (m

m H

g):

Neg

ligib

le.

Vapo

r Den

sity

(Air

= 1)

: N�A

Evap

orat

ion

rate

(But

yl a

ceta

te =

1):

N�A

Visc

osity

: N�A

Che

mic

al s

tabi

lity:

Sta

ble

Haz

ardo

us p

olym

eriz

atio

n: W

ill n

ot o

ccur

.C

ondi

tions

to a

void

: H

igh

tem

pera

ture

cau

ses

deco

mpo

sitio

n to

sod

ium

car

bona

te, w

ater

and

car

bon

diox

ide.

Inco

mpa

tibili

ties

with

oth

er m

ater

ials

: R

eact

s w

ith a

cids

to y

ield

aci

d sa

lts, w

ater

and

car

bon

diox

ide.

Haz

ardo

us d

ecom

posi

tion

prod

ucts

: G

aseo

us c

arbo

n di

oxid

e.

Boi

ling

poin

t: D

ecom

pose

s to

rele

ase

CO

2Fr

eezi

ng /

Mel

ting

poin

t: 1

09°C

(228

°F)

Dec

ompo

sitio

n te

mpe

ratu

re:

N�A

So

lubi

lity:

9.6

gra

ms

per 1

00 m

l at 2

0°C

Spec

i¿c

grav

ity (H

2O =

1):

2.1

6 at

20°

CPe

rcen

t vol

atile

(%):

N�A

Mol

ecul

ar fo

rmul

a:

NaH

CO

3M

olec

ular

wei

ght:

84.

01

MS

DS

No.

: SS

0270

Rea

d la

bel o

n co

ntai

ner b

efor

e us

ing.

Do

not w

ear c

onta

ct le

nses

whe

n w

orki

ng w

ith c

hem

ical

s. K

eep

cont

aine

r tig

htly

clo

sed.

For

labo

rato

ry u

se o

nly.

Not

for d

rug,

food

or h

ouse

hold

use

. K

eep

out o

f rea

ch o

f chi

ldre

n.H

andl

ing:

Use

with

ade

quat

e ve

ntila

tion.

Avo

id c

onta

ct w

ith e

yes,

ski

n an

d cl

othi

ng.

Avoi

d in

gest

ion.

Do

not

inha

le d

usts

. W

ash

thor

ough

ly a

fter h

andl

ing.

Rem

ove

and

was

h cl

othi

ng b

efor

e re

use.

St

orag

e: S

tore

in a

coo

l, dr

y, w

ell-v

entil

ated

are

a.

Non

e lis

ted.

Aldon

Corporation

221

Roc

hest

er S

treet

Avon

, NY

1441

4(5

85) 2

26-6

177

Documents

AP® INVESTIGATION #5 - Wikispaceshinton.harvardmedk12.wikispaces.net/file/view/Ward's... · Objectives of the lab as described in AP Biology Investigative Labs: An Inquiry-Based