Chromatography of Plant Pigments 3

 

 

Chromatography of Plant Pigments

 

 

Introduction:

 

Can chromatography be used to separate mixtures of chemical substances? The purpose of this experiment is to answer this question. In paper chromatography, a liquid sample flows down a vertical strip of absorbent paper, on which the components of a mixture are deposited in specific directions and locations. Chromatography is a tool used to examine and separate mixtures of chemical substances. Chromatography is essential to the separation of pure substances from complex mixtures. Separation results in a chromatographically pure substance. Chromatography allows you to determine the properties of chemical substances.

The relationship between the chromatography paper, mixture, and the solvent is very important in all chromatographic separations. The solvent has to dissolve the mixture that should be separated. The paper must also absorb the components of the mixtures selectively and reversibly. The substances making up the mixture must be evenly dispersed in the water. Chromatography is a simple and inexpensive tool for separating and identifying chemical mixtures if all these things are done.

 

Hypothesis:

 

Paper can be used to separate mixed chemicals.

 

Materials:

 

The materials used in this lab are filter paper, test tube, rubber stopper, paper clip, metric ruler, black felt-tip pen, pencil, calculator, and water.

 

Methods:

 

First, bend a paper clip so that it’s straight with a hook at one end. Push the straight end of the paper clip into the bottom of a cork stopper. Then, hang a thin strip of filter paper on the hooked end of the paper clip and insert the paper strip into the test tube. The paper should not touch the sides and should almost touch the bottom of the test tube. Next, remove the paper strip from the test tube. Now draw a solid 5-mm-wide band about 25 mm from the bottom of the paper, using a black felt tip pen. After this, use a pencil to draw a line across the paper strip 10 cm above the black band. Then, put the filter paper back into the test tube with the bottom of the paper in the water and the black band above the water. Observe what happens as the liquid travels up the paper and record the changes you see. When the solvent has reached the pencil line, remove the paper from the test tube. Let the paper dry on the desk. Finally, with a metric ruler, measure the distances from the starting point to the top edge of each color. Record the data in a data table and calculate a ratio for each color by dividing the distance, the color traveled by the distance the solvent traveled.

 

Results:

 

The results of the chromatography experiment are shown in a chart and a graph.

 

Color of ink (list in order) Distance traveled by each color (mm) Distance solvent traveled (mm) Ratio traveled = distance color moved divided by distance solvent moved
Yellow 70 108 0.65
Orange  

85

 

108

 

0.79

Pink 95  

108

 

0.88

Violet 102  

108

 

0.94

Blue 108  

108

 

1.00

 

 

 

Questions:

1. How many colors separated from the black ink? Five colors separated from the ink: yellow, orange, pink, violet, and blue.

 

2. What served as the solvent for the ink? Water served as the solvent for the ink.

 

3. As the solvent traveled up the paper, which color of ink appeared first? Dark blue appeared first.

 

4. List the colors in order from top to bottom that separated from the black ink? The colors separated in the order of: blue, violet, pink, orange, and yellow.

 

5. In millimeters, how far did the solvent travel? The solvent traveled 108 mm.

 

6. From your results, what can you conclude is true about black ink? Black ink is a mixture of several different colors.

 

7. Why did the inks separate? The inks separated because black ink is a mixture of different pigments that are soluble in water, have different molecular characteristics, and travel different distances.

 

8. Why did some inks move a greater distance? Some inks move a greater distance because molecules in ink have different characteristics, like how readily they are absorbed by paper. This means that the ink least readily absorbed by paper will travel farthest from the starting mark and the ink most readily absorbed by paper will be the closest to the starting mark. All of the different color inks that were separated were different in how readily they are absorbed by paper.

Error Analysis:

 

There are a few errors that could have changed the results. First, there could be inaccurate measurements of how far every color traveled or how far the water traveled up the filter paper. Another error could occur when calculating the ratio traveled, Rf value. Also, a longer test tube could have been used by different groups which would make the filter strip longer. This means that a group could have detected another color because they had more room on their filter paper. This also could have affected the ratios. Finally, the groups could have put different amounts of black ink on the filter paper.

 

Conclusion:

 

The hypothesis that paper can be used to separate mixed chemicals was correct. The different colored inks mixed together give the black its color. The five colors that separated from the black ink were blue, violet, pink, orange, and yellow. Blue appeared first and then was followed by violet, pink, orange, and yellow. The colors separated the way they did because they have different molecular characteristics, like how readily they were absorbed by the paper and their solubility in water. Blue was most readily absorbed by the paper and soluble by water, while yellow was the least.

BACK

 

Charles Robert Darwin

 


Charles Robert Darwin
Darwin and Evolution

 

 

History of Evolution:

  • Plato & Aristotle believed species were fixed & could be arranged according to their complexity
  • In the mid eighteenth century, Carolus Linnaeus developed a system of classification that described the fixed features of species and revealed God’s divine plan
  • George Cuvier, in the eighteenth century to explain changes in the fossil record, proposed that a whole series of catastrophes (extinctions) and re-populations from other regions had occurred giving the appearance of change over time
  • Prior to Darwin, it was thought that the world was young & species did not change
  • In 1831, at the age of 22, Charles Darwin accepted a naturalist position aboard the ship HMS Beagle & began a five-year voyage around the world
  • Darwin’s many observations led him to the idea that species slowly change

C. Late Eighteenth Century Contributions

1.
a. George Cuvier (1769-1832), a distinguished French vertebrate zoologist, was the first to use
comparative anatomy to develop a system of classifying animals.
b. He founded the science of paleontology, the study of fossils, and suggested that a single fossil bone
was all he needed to deduce the entire anatomy of an animal.
c. .
d. Cuvier was also a staunch advocate of special creation and fixity of species; this presented him with
a serious problem when geological evidence of a particular region showed a succession of life forms
in the earth’s strata.
e. Catastrophism is the term applied to Cuvier’s explanation of fossil history, the belief held by Cuvier
that catastrophic extinctions occurred, after which repopulation of surviving species took place,
giving an appearance of change through time.
2. Lamarck’s Theory of Evolution
a. Lamarck (1744-1829) was first to state that descent with modification occurs and that organisms
become adapted to their environments.
b. Lamarck was an invertebrate zoologist and held ideas different from Cuvier.
c. Unfortunately, he saw the drive for perfection as inherent in all living things.
d. Inheritance of acquired characteristics was the Lamarckian belief that organisms become adapted to
their environment during their lifetime and pass on these adaptations to their offspring.
e. He believed the increasing complexity of life forms in strata is the result of a natural tendency toward
perfection inherent in all living things.
f. Experiments fail to uphold Lamarck’s inheritance of acquired characteristics; molecular mechanism of
inheritance show phenotypic changes do not result in genetic changes that can be passed on.

18.2. Darwin’s Theory of Evolution

A. Darwin’s Background

1. His nature was too sensitive to pursue medicine; he attended divinity school at Cambridge.
2. He attended biology and geology lectures and was tutored by the Reverend John Henslow.
3. Henslow arranged his trip on the HMS Beagle; Darwin was an observant student of nature.

B. Geology and Fossils

1. His study of geology and fossils caused him to concur with Lyell that the observed massive geological
changes were caused by slow, continuous processes.
a. In his book Principles of Geology, Charles Lyell presented arguments to support a theory of geological
change proposed by James Hutton.
b. In contrast to catastrophists, Hutton proposed that the earth was subject to slow but continuous
geological processes (e.g., erosion and uplifting) that occur at a uniform rate.
c. Darwin took Lyell’s book on the voyage of the HMS Beagle.
2. Fossil Evidence
a. The Argentina coast had raised beaches; he witnessed earthquakes raising the earth several feet.
b. Marine shells occurred far inland and at great heights in the Andes.
c. Fossils of huge sloths and armadillo-like animals suggested modern forms were descended from
extinct forms with change over time.

C. Biogeography

1. Biogeography is the study of the geographic distribution of life forms on earth.
2. Darwin’s comparison of the animals of South America and the Galapagos Islands caused him to conclude
that adaptation to the environment can cause diversification, including origin of new species.
3. Patagonian hares replaced rabbits in the South American grasslands.
4. The greater rhea found in the north was replaced by the lesser rhea in the south.
5. The Galapagos Islands
a. These volcanic islands off the South American coast had fewer types of organisms.
b. Island species varied from the mainland species, and from island-to-island.
c. Each island had a variation of tortoise; long and short necked tortoises correlated with different vegetation.
d. Darwin’s Finches
1) Finches on the Galapagos Islands resembled a mainland finch but there were more types.
2) Galapagos finch species varied by nesting site, beak size, and eating habits.
3) One unusual finch used a twig or thorn to pry out insects, a job normally done by a woodpecker.
4) The finches posed questions to Darwin: did they descend from one mainland ancestor, did islands
allow isolated populations to evolve independently, and could present-day species have resulted
from changes occurring in each isolated population?

D. Natural Selection and Adaptation

1. Darwin decided adaptations develop over time; he sought a mechanism by which adaptations might arise.
2. Natural selection was proposed by both Alfred Russel Wallace and Darwin as a driving mechanism of
evolution caused by environmental selection of organisms most fit to reproduce, resulting in adaptation.
3. Because the environment is always changing, there is no perfectly-adapted organism.
4. Preconditions for natural selection
a. The members of a population have random but heritable variations.
b. In a population, many more individuals are produced each generation than an environment can support.
c. Some individuals have adaptive characteristics that enable them to survive and reproduce better.
5. Consequences of natural selection
a. An increasing proportion of individuals in succeeding generations have the adaptive characteristics.
b. The result of natural selection is a population adapted to its local environment.
6. Natural selection can only utilize variations that are randomly provided; therefore, there is no directedness
or anticipation of future needs.
7. Extinction occurs when previous adaptations are no longer suitable to a changed environment.

E. Organisms Have Variations

1. In contrast to the previous world-view, variations are highly significant.
2. Darwin suspected, but did not have today’s evidence, that variation is completely random.
3. New variations are as likely to be harmful as helpful.
4. Variations that make adaptation possible are those that are passed on generation to generation.
5. Darwin could not state the cause of variations because genetics was not yet established.

F. Organisms Struggle to Exist

1. Darwin and Wallace both read an essay by Thomas Malthus, a clergyman and socio-economist.
2. Malthus proposed that human populations outgrow resources and death and famine were inevitable.
3. Darwin applied this to all organisms; resources were not sufficient for all members to survive.
4. Therefore, there is a constant struggle for existence; only certain members survive and reproduce.

G. Organisms Differ in Fitness

1. Organisms whose traits enable them to reproduce to a greater degree have a greater fitness.
a. Fitness is a measure of an organism’s reproductive success.
b. Black western diamondback rattlesnakes are more likely to survive on lava flows; lighter-colored
rattlesnakes are more likely to survive on desert soil.
2. Darwin noted that humans carry out artificial selection.
a. Early humans likely selected wolf variants; consequently, desirable traits increase in frequency in
subsequent generations and produced the varieties of domestic dogs.
b. Many crop plant varieties can be traced to a single ancestor.
c. In nature, interactions with the environment determine which members reproduce more.
d. Evolution by artificial or natural selection occurs when more fit organisms reproduce and leave more
offspring that the less fit.

H. Organisms Become Adapted

1. An adaptation is a trait that helps an organism be more suited to its environment.
2. Unrelated organisms living in the same environment often display similar characteristics.
3. Because of differential reproduction, adaptive traits increase in each succeeding generation.

I. On Origin of Species by Darwin

1. After the HMS Beagle returned to England in 1836, Darwin waited over 20 years to publish.
2. He used the time to test his hypothesis that life forms arose by descent from a common ancestor and
that natural selection is a mechanism by which species can change and new species arise.
3. Darwin was forced to publish Origin of Species after reading a similar hypothesis by Alfred Russel Wallace.

18.3. Evidence for Evolution

A. Common Descent Adapted

1. The hypothesis of common descent is supported by many lines of evidence.
2. The more varied the evidence, the more certain it becomes.
3. Darwin synthesized much of the current data but biochemical research was yet to come.

B. Fossils Evidence

1. The fossil record is the history of life recorded by remains from the past.
2. Fossils are at least 10,000 years old and include skeletons, shells, seeds, insects trapped in amber,
and imprints of leaves.
3. The fossil record traces history of life and allows us to study history of particular organisms.
4. Fossil evidence supports the common descent hypothesis; fossils can be linked over time because they
reveal a similarity in form, despite observed changes.
5. Transitional forms reveal links between groups.
a. Caudipteryx is between dinosaurs and birds.
1) This Chinese fossil shows some dinosaurs had feathers on arms, tail and probably body.
2) Advantages during running and escape gave rise to birds once lift-off occurred.
b. Eustheopteron is an amphibious fish.
c. Seymouria is a reptile-like amphibian.
d. Therapsids were mammal-like reptiles.
6. The fossil record allows us to trace the history of the modern-day horse Equus.
a. Earliest fossils show an ancestral Hyracotherium the size of a dog, with cusped low-crowned molars,
four toes on each front foot, three on each hind foot — all adaptations for forest living.
b. When forests were replaced by grasslands, the intermediates were selected for durable grinding teeth,
speed, etc. with an increase in size and decrease in toes.
c. Living organisms resemble most recent fossils in the line of descent; underlying similarities allow us
to trace a line of descent over time.

C. Biogeographical Evidence

1. Biogeography studies the distribution of plants and animals worldwide.
2. Distribution of organisms is explained by related forms of evolving in one locale and spreading to
other accessible areas.
a. Darwin observed South America had no rabbits; he concluded rabbits originated elsewhere.
b. Biogeography explains the many finch species on the Galapagos Islands but not the mainland.
3. Physical factors, such as the location of continents, determine where a population can spread.
a. Cacti are restricted to North American deserts and euphorbia grow in African deserts.
b. Marsupials arose when South America, Antarctica, and Australia were joined; Australia separated
before placental mammals arose, so only marsupials diversified in Australia.

D. Anatomical Evidence

1. Organisms have anatomical similarities when they are closely related because of common descent.
a. Homologous structures in different organisms are inherited from a common ancestor.
b. Analogous structures are inherited from a unique ancestors and have come to resemble each other
because they serve a similar function.
c. Vertebrate forelimbs contain the same sets of bones organized in similar ways, despite their dissimilar functions.
2. Vestigial Structures are remains of a structure that was functional in some ancestor but is no longer
functional in the organism in question.
a. Most birds have well-developed wings; some bird species have reduced wings and do not fly.
b. Humans have a tailbone but no tail.
c. Presence of vestigial structures is explained by the common descent hypothesis; these are traces of
an organism’s evolutionary history.
3. Embryological development reveals a unity of plan.
a. During development, all vertebrates have a post-anal tail and paired pharyngeal pouches.
1) In fishes and amphibian larvae, the pouches become gills.
2) In humans, first pair of pouches becomes a cavity of middle ear and auditory tube; second pair
becomes tonsil, while third and fourth pairs become thymus and parathyroid glands.
3) Above features are explained if fishes are ancestral to other vertebrate groups.

E. Biochemical Evidence

1. Almost all living organisms use the same basic biochemical molecules, e.g., DNA, ATP, and many
identical or nearly identical enzymes.
2. Organisms utilize the same DNA triplet code and the same 20 amino acids in their proteins.
3. Many organisms share same introns and types of repeats, which is remarkable since there is no obvious
functional reason why these components need to be so similar.
4. This is substantiated by analysis of degree of similarity in amino acids for cytochrome c among organisms.
5. These similarities can be explained by descent from a common ancestor.
6. Life’s vast diversity has come about by only a slight difference in the same genes.

F. Because it is supported by so many lines of evidence, evolution is no longer considered a hypothesis.

1. Evolution is one of the great unifying theories of biology.
2. In science, theory is reserved for those conceptual schemes that are supported by a large number of
observations or a large amount of experimental evidence and have not been found lacking.

 

evolution–process of change through time

Evolution includes the change in characteristics of populations through generations.
Thus, existing life forms have evolved from earlier life forms.

Evolutionary theory is a unifying principle for the biological sciences.
It provides an explanation for the differences in structure, function, and behavior
among life forms.

Through radioactive dating with uranium, geologists
estimate the age of the earth at about 4.6 billion years. (It is assumed that the earth
is at least as old as the oldest rocks and minerals composing its crust.)

fossils: direct or indirect remains of organisms preserved in media such as sedimentary rock, amber, ice, or tar

Fossils have been found that indicate organisms existed well over 3 billion years ago. These organisms were simple, single-celled organisms.

Law of Superposition: the higher up you go in an undisturbed rock stratum, the
younger the rock layers become

** Upper, undisturbed strata generally contain fossils of more complex organisms,
whereas, the lower strata contain fossils of simpler life forms.
(Tendency toward increasing complexity over time.)

** When comparing fossils in undisturbed strata, fossils can be found in
upper strata which, although different from fossils in lower strata, resemble
those fossils. This suggests links between modern forms and older forms, as
well as divergent pathways from common ancestors.

adaptations–changes in organisms which make them better suited to their environment

Gene mutations can be caused by such agents as radiation and chemicals.  When they occur in sex cells, the mutations can be passed on to the offspring; if they occur in other cells, they can be passed on to body cells only.  The experiences an organism has during its lifetime can affect its offspring only if the genes in its own sex cells are changed by the experience.

biological adaptation — changes in structures, behaviors, or physiology that enhance survival and reproductive success in a particular environment

Theory of Natural Selection

Darwin–(1859) Book–“On Origin of Species”

–stated his theory of Natural Selection

— Natural selection and its evolutionary consequences provide a scientific explanation of the fossil record of ancient life-forms, as well as the molecular and structural similarities observed among the diverse species of living organisms.

(Theory of Natural Selection)

1. Overproduction — Within a population more offspring are born than can possibly survive.

2. Competition — since the number of individuals in a population tends to remain constant from generation to generation, a struggle for survival is suggested

3. Survival of the Fittest — The individuals who survive are the ones best adapted to exist in their environment due to the possession of variations that maximize their fitness.

4. Reproduction — Variations assist or hinder individuals in their struggle for survival.
The best adapted individuals survive and reproduce, passing on  the favorable variations to their offspring.

5. Speciation — as time and generations continue, adaptations are perpetuated in individuals and new species may evolve from a common ancestor.

adaptive variations–those variations which assist an organisms survival

(Weaknesses of Darwin’s Theory)

1. Darwin didn’t explain how variations arose.
2. He did not distinguish between hereditary and environmental variations.
3. Darwin believed that both environmental and hereditary variations were inherited.

Some Sources of Variation in Modern Natural Selection Theory

1. The genetic basis for variation within a species is provided by mutations and sexual reproduction. (crossing over and recombination)

2. Mutations are spontaneous and provide the raw material for evolution.

(Modern Natural Selection Theory)

1.)  All species of the potential to increase in numbers.

2.)  There is a finite amount of resources for any species.   Species tend to make too many
organisms for these resources.

3.)  Species will show genetic variability due to mutation, crossing over, and
genetic recombination (during fertilization) of genes.

4.)  The scarce finite resources of the environment will select those offspring better able to survive and leave offspring.

Variation within a species increases the likelihood that at least some members of the species will survive under changed environmental conditions.

** Traits which are beneficial to the survival of an organism in a particular environment tend to be retained and passed on, and therefore, increase in frequency within a population.

** Traits which have a low survival value to organisms tend to diminish in
frequency from generation to generation.

** If environmental conditions change, traits that were formerly associated with a
low survival value may, in a changed environment, have greater survival value and
increase accordingly.

(Examples of Evolution in Modern Times)

1. Peppered moth — light colored vs. dark colored (industrialization influence) Manchester, England
2. Insect resistance to insecticides. (Resistance is not in response to the insecticide. The insecticide acts as a selecting agent.)
3. Bacterial resistance to antibiotics.

** Evolutionary factors operate on population — but not on individual organisms.

speciation: formation of two or more different  species from one original population

Evolution does not necessarily mean long term progress is going to go in a certain direction.   Evolutionary changes often appear to be like the growth of a bush: Some branches survive from the beginning with little or no change, many die out altogether, and others branch out repeatedly, sometimes giving rise to more complex organisms.

Note the divergence of the various groups from a common ancestor and the fact that some branches became extinct.

Extinction of a species occurs when the environment changes and the adaptive characteristics of a species are insufficient to allow its survival.   Fossils indicate that many organisms that lived long ago are extinct.   Extinction of a species is common; most of the species that have lived on earth no longer exist.

The SPECIES is the most fundamental unit of classification.

The purpose of biological classification is to show how organisms are related.  Organisms are branched into hierarchies or groups based on structural similarities and evolutionary relationships.

Small differences between parents and offspring can accumulate in successive generations so that descendants become very different from their ancestors.

**  The degree of kinship between organisms or species can be estimated from the similarity of their
DNA sequences; this similarity often closely matches organisms’ or species’ classification based on anatomical similarities.

Origin of Life

1.)  3 to 4 billion years ago — it is thought the first primitive single-celled life appeared on earth

2.) These original unicellular organisms added of carbon dioxide to the environment.

3.) Some autotrophs evolved a means of using the carbon dioxide added to do photosynthesis

4.) Autotrophic activity added free oxygen to the atmosphere. Some autotrophs
and heterotrophs evolved mechanisms by which they used this oxygen to
derive their energy. (aerobic respiration)

5.) About a billion years ago, increasingly complex multicellular organisms began to evolve.

**  The great diversity of organisms is the result of billions of years of evolution that has filled available niches with life-forms.

Chromatography of Simulated Plant Pigments

 

Chromatography of Simulated Plant Pigments

 

Introduction
    This experiment is conducted to investigate the components Plant Pigments separating visibly. There are a couple of different types of components in plant pigments, and they became clearly visible during this lab. The most important and abundant chemical pigment found in plants is chlorophyll. This pigment exists in two forms; chlorophyll a and chlorophyll b. Chlorophyll absorbs two main colors from light quite well. These are blue, and red. The chlorophyll reflects green light very well, however, the two different types of chlorophyll have their maximum absorption at different wavelengths of light. Chlorophyll a, being the main photosynthetic pigment, has a primary purpose to convert light energy to chemical energy used by the plant itself. Chlorophyll b absorbs light in a region of the spectrum apart from the dominant chlorophyll, and transfers the energy it produces to chlorophyll a. Along with chlorophyll b in transferring their energy produced to the dominant chlorophyll, two other pigments that are found in plants are carotenes and xanthophylls, which are orange and yellow respectively. Since chlorophyll is such a dominant pigment in green plants, this domination hides the color of the carotenes and xanthophylls in the leaves. This causes most plant leaves to appear green most of the time. During the autumn, however, the chlorophyll starts to break down, causing the carotenes and xanthophylls to show their bright red, orange and yellow colors.
These brilliant colors can be separated another way. This different technique, known as paper chromatography, separates mixtures in a liquid into individual components. The technique is based on the fact that each substance in a mixture has a specific affinity for a solid surface and a specific solubility in different solvents. By this method, the solid surface is the cellulose fibers in the chromatography paper, and the solvent is the solution that was placed in the bottom of the developing chamber.
This separation takes place through a process of absorption and capillary action. Just a small drop of the mixture, in this case plant pigment to be separated, is placed at the bottom of the strip of chromatography paper. The chromatography paper is then placed in the developing chamber with a solvent, which wicks up the paper, pulling the solvent up the paper by capillary action, and the mixture of pigments is dissolved as the solvent passes over it. The different components of the mixture move upward at different rates. A compound with greater solubility will travel farther than one with less solubility. The pigments then show up as color streaks on the chromatography paper. These substances have formed a pattern called a chromatogram on the chromatography paper.
The Rf values for each pigment is calculated to establish the relative rate of migration for each pigment. This value represents the ratio of the distance a pigment traveled on the chromatogram relative to the distance the solvent front moved.
Scientists use the Rf value of a sample to identify the molecule. Any molecule in a given solvent matrix system has a uniquely consistent Rf value. The formula for this value is as follows:

Rf = Distance each pigment traveled ¸ Distance solvent front traveled

 

Hypothesis
    Using paper chromatography, the pigments that give a leaf its color can be separated and observed to determine the Rf value of each pigment and their function during photosynthesis.

 

Materials
For this experiment the following items are used — one chromatography reaction chamber, one paper chromatography strip, one capillary pipette, a pencil and paper, calculator, ruler, 50 ml beaker, colored pencils, approximately 10 ml of solvent depending on the size of the reaction chamber, scissors, and simulated plant pigment.

 

Procedure
Use scissors to cut the bottom of the chromatography paper to a tapered end. Measure the strip and cut the length to equal slightly longer than the reaction chamber. Draw a faint pencil line at the bottom of the tapered end and use a capillary pipette to add some simulated plant pigment to this line. Add 5-10 ml of solvent to the reaction chamber. Extend the chromatography strip through the slit in the lids of the reaction chamber and carefully lower the strip into the chamber so the tapered end is in the solvent and the pencil line is above the solvent level. Make sure the strip does not touch the walls of the chamber and do not bump the chamber as the pigments begin to separate. After the pigments have completely separated and the solvent front has reached the top of the chamber, remove the strip and mark the solvent front with a pencil line before it evaporates. Measure and record the distance the solvent and each pigment traveled. Use a calculator to determine the Rf values for each pigment.

 

Data

 

Table 1

Band # Pigment Color Migration distance (mm) Rf value
1 Carotene Orange 59mm .94
2 Xanthophyll Yellow 56mm .89
3 Chlorophyll a Light green 29mm .46
4 Chlorophyll b Dark green 14mm .22
Solvent 63mm

Questions
1. Describe what happened to the original spot of simulated plant pigments?
  The solvent separated  the original spot by wicking up the solvent while dissolving the various pigments in the spot.
2. List some other uses of chromatography?  Chromatography can be used to separate various mixtures of subtances, liquids and gases.
3. Which of the 4 pigments migrated the furthest and why?  carotene ( orange) because it was the most soluble in the solvent
4. Which type of chlorophyll was the most soluble?  chlorophyll a
5. Explain why leaves change color in the fall?  In Autumn, chlorophyll starts to break down which allows the other brilliant plant pigment colors to show. These pigments include the red, orange, and yellow colors.
6. What is the function of plant pigments in photosynthesis?  Plant pigments trap light energy and convert it into chemical energy that can be used by the plant to make glucose or sugar.

Error Analysis
The chromatography paper touched the sides of the chamber during the waiting time which caused the migration to go slightly to the side instead of straight to the top. Also the strip was bent at the top so there could have been a slight error in measuring the migration of the solvent  front.

Conclusion
Paper chromatography proved to be an accurate method of separating and observing the various colors of plant pigments. The pigments dissolved in the solvent and migrated upward. The colors were observed and their migration distances measured & recorded. The
Rf value of each pigment was determined by dividing its migration by the migration of the solvent.  It was determined that 4 pigments were present in the original spot — carotene, xanthophyll, chlorophyll a, and chlorophyll b. Carotene was the most soluble, while chlorophyll b was the least soluble.

Chemistry Quiz

Name: 

Chemistry Quiz

 

True/False
Indicate whether the sentence or statement is true or false.
1.
The types of particles that are located in the nucleus of an atom are protons and neutrons, and the types of particles that are located in the energy levels surrounding the nucleus are electrons.
2.
The atomic number of carbon is 6, so it must contain 12 electrons.
3.
Most elements are stable if their outermost energy level contains an even number of electrons.
4.
Atoms in a gas move more rapidly than atoms in a liquid or a solid do.
5.
The products of an endergonic chemical reaction possess more energy than the reactants from which they are produced.
6.
The amount of energy needed to cause a chemical reaction to start is called activation energy.
7.
When an enzyme binds with its substrate, the activation energy needed for the chemical reaction to occur is raised.
8.
Enzymes speed up a chemical reaction by increasing the activation energy of the reaction.
9.
Free hydrogen ions can react with water molecules and form a positively charged ion, the hydronium ion.
10.
Buffers can neutralize acids, but they do not affect bases.
 

Multiple Choice
Identify the letter of the choice that best completes the statement or answers the question.
11.
The smallest particle of matter that can retain the chemical properties of carbon is
a.
a carbon molecule.
c.
a carbon atom.
b.
a carbon macromolecule.
d.
an element.
12.
The bond formed when two atoms share a pair of electrons is called a
a.
hydrogen bond.
c.
covalent bond.
b.
nonpolar bond.
d.
water bond.
13.
Sharing of electrons in the outer energy levels of two atoms
a.
results in ion formation.
b.
occurs in covalent bonds.
c.
only occurs if both are atoms of the same element.
d.
is found only among carbon atoms.
14.
An atom that has gained or lost electrons is called a(n)
a.
molecule.
c.
ion.
b.
nucleon.
d.
element.
15.
Which of the following statements most accurately describes the difference between an ionic bond and a covalent bond?
a.
Atoms held together by ionic bonds separate when placed in water while atoms held together by covalent bonds do not separate in water.
b.
Ionic bonds hold together atoms of two different types, while covalent bonds hold together atoms of the same type.
c.
Electrons are exchanged between atoms held together by an ionic bond, but they are shared between atoms held together by a covalent bond.
d.
Ionic bonds form between atoms that carry opposite charges, while covalent bonds form between uncharged atoms.
16.
A reaction in which the products have less energy than the reactants is
a.
an endergonic reaction.
c.
a filamentous reaction.
b.
an exergonic reaction.
d.
impossible.
      chemistry_quiz_files/i0190000.jpg
17.
Refer to the graph above. Reaction “1” in the graph
a.
is an energy-storing reaction.
b.
requires a greater activation energy than Reaction “2.”
c.
may use the same initial reactant condition needed to form Product “B.”
d.
All of the above
18.
Refer to the graph above. Which of these statements is true regarding the graph?
a.
Reaction “2” occurs faster than Reaction “3” because Reaction “2” requires more energy than Reaction “3.”
b.
The difference in the graphs shown for Reaction “2” and Reaction “3” is due to a difference in the activation energy needed for these reactions.
c.
Reactant “A” contains more energy at the beginning of the reaction than Product “C” has after the reaction.
d.
All of the above
19.
Changing the course or pathway of a chemical reaction so that it requires less activation energy
a.
is a violation of the laws of nature.
b.
requires higher temperatures than those found within cells.
c.
occurs only when reactants are quickly added to the reaction mixture.
d.
is accomplished by the action of catalysts on reactants.
20.
Oxidation-reduction reactions are important in organisms because they
a.
allow the passage of energy from molecule to molecule.
b.
prevent nuclear reactions from occurring.
c.
allow the creation and destruction of energy.
d.
None of the above; oxidation-reduction reactions do not occur in living organisms.

 

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Chromatography Plant Pigments

 

Chromatography of Plant Pigments

INTRODUCTION:

Chlorophyll often hides the other pigments present in leaves. In Autumn, chlorophyll breaks down, allowing xanthophyll and carotene, and newly made anthocyanin, to show their colors.
The mix of pigments in a leaf may be separated into bands of color by the technique of paper chromatography. Chromatography involves the separation of mixtures into individual components. Chromatography means “color writing.” With this technique the components of a mixture in a liquid medium are separated. The separation takes place by absorption and capillarity. The paper holds the substances by absorption; capillarity pulls the substances up the paper at different rates. Pigments are separated on the paper and show up as colored streaks. The pattern of separated components on the paper is called a chromatogram.

PRELAB PREPARATION:

Gather leaves from several different plants. CAUTION: Avoid poisonous plants. Autumn leaves from deciduous trees are especially interesting. Sort the leaves by kind (maple, etc.) and color. Review a diagram of a plant cell . Find the grana and the chloroplasts of the cell.

MATERIALS:

Safety goggles
Chromatography solvent (92 parts Petroleum ether to 8 parts acetone)
Chromatography paper (or filter paper) about 1 cm x 15 cm
Ethyl alcohol
Fresh spinach
Test tube
Test tube rack
Scissors and Ruler
Fresh leaves of plants
Glass stirring rod
Paper clip
Cork (to fit test tube)
Mortar and pestle
Sand (optional)
10-ml Graduated cylinder

PROCEDURE:

Leaves should be grouped by kind (maple, etc.) and color. Work with a spinach leaf and with one or more other types. CAUTION: Chromatography solvents are flammable and toxic. Have no open flames; maintain good ventilation; avoid inhaling fumes.

1. Cut a strip of filter paper or chromatography paper so that it just fits inside a 15-cm (or larger) test tube. Cut a point at one end. Draw a faint pencil line as shown in figure 1. Bend a paper clip and attach it to a cork stopper. Attach the paper strip so that it hangs inside the tube, as shown. The sides of the strip should not touch the glass.

2. Tear a spinach leaf into pieces about the size of a postage stamp. Put them into a mortar along with a pinch or two of sand to help with grinding. Add about 5 ml ethyl alcohol to the leaf pieces. Crush leaves with the pestle, using a circular motion, until the mixture is finely ground. The liquid in which the leaf pigments are now for paper chromatography dissolved is called the pigment extract.

3. Use a glass rod to touch a drop of the pigment extract to the center of the pencil line on the paper strip. Let it dry. Repeat as many as 20 times, to build up the pigment spot. NOTE: You must let the dot dry after each drop is added. The drying keeps the pigment dot from spreading out too much.

4. Pour 5 ml chromatography solvent into the test tube. Fit the paper and cork assembly inside. Adjust it so that the paper point just touches the solvent (but not the sides of the tube). The pigment dot must be above the level of the solvent. Watch the solvent rise up the paper, carrying and separating the pigments as it goes. At the instant the solvent reaches the top, remove the paper and let it dry. Observe the bands of pigment. The order, from the top, should be carotenes (orange), xanthophylls (yellow), chlorophyll a (yellow-green), chlorophyll b (blue-green), and anthocyanin (red). Identify and label the pigment bands on the dry strip. Write the species of leaf on the strip as well.
Record the species, external color, and chromatogram pigments in the DATA TABLE of your report sheet.

5. Each pigment has an Rf value, the speed at which it moves over the paper compared with the speed of the solvent.

Rf = Distance moved by the pigment / Distance moved by the solvent

Measure the distance in cm from the starting point (pencil line) to the center of each pigment band. Then measure the entire distance traveled by the solvent. Remember, the starting point for the solvent is also the pencil line and the ending point for the solvent is the top edge of the paper. Do the required divisions and record your Rf values in the DATA TABLE of your report sheet.

6. Wash the mortar and pestle thoroughly, using a little alcohol to remove any remaining pigment.

7. Repeat steps 1 through 6 for each species.

DATA TABLE:

Chromatography Data

Leaf Type (species) External color Chromatogram Pigments
Colors from the Top Pigment Names Rf Values