Ink Chromatography

Chromatography of Inks

Introduction:

One of the main jobs of biochemists is to unravel the complexities of chemical compounds and reduce them to their individual components.  The term chromatography comes from two Greek words, “chromat” meaning color and the word “graphon” meaning to write.  Separation of the components of chemical compounds can be done by using several methods. Liquids can be separate by High Performance liquid Chromatography (HPLC), while the components of gases are separated by Gas Chromatography.  Chromatography is a method for analyzing complex mixtures (such as ink) by separating them into the chemicals from which they are made. Chromatography is used to separate and identify all sorts of substances in police work. Drugs from narcotics to aspirin can be identified in urine and blood samples, often with the aid of chromatography.

Chromatography was first used to separate pigments (colors) in leaves, berries, and natural dyes. Paper chromatography is a technique used to separate, isolate, and identify chemical components of a compound. In paper chromatography, the solid surface is the cellulose fibers in the chromatography paper.  A solvent or developer (water, alcohol, or acetone) is placed in the bottom of the chromatography chamber. The paper acts as a wick to pull the solvent up the paper. The solvent front will “wick” up the chromatography paper by capillary action.  A minute drop of the ink or chemical mixture to be separated is placed near the bottom of the strip of chromatography paper, but slightly above the level of the solvent in the chamber.  As the solvent passes over the drop of ink, the components of the ink dissolve in the solvent. Because the components of the ink do not all dissolve at the same rate, as the components of the mixture move upward, they show up as colored streaks.  The separated substances on the chromatography paper form a color pattern called a chromatogram.

To determine the rate of migration for each pigment or component of the ink, the Rf value for each pigment must be calculated. The Rf value represents the ratio of the distance a pigment moved on the chromatogram relative to the  distance the solvent front moved. Each pigment or compound will have a unique Rf value that scientists can use to identify the substance. The Rf value is calculated using the following formula:

Rf = distance traveled by the compound / distance traveled by the solvent

Objective:

Use the process of paper chromatography to separate the pigments in various markers and then determine the Rf value for each color on your chromatogram.

Materials:

Plastic vials, paper clips, markers in assorted colors, chromatography paper, scissors, pencil

Procedure:

  1. Obtain chromatography vials and chromatography strips, and different color markers so that each person in the group will have two chromatograms.
  2. Cut one end of the chromatography strip to a point. The bottom of the point will mark the starting point for movement of the solvent (H2O).
  3. About 2.0 centimeters from the bottom of the strip, draw a faint horizontal line with pencil. This will mark the starting point for measuring the migration distance of each color.
  4. Using a different color marker for each strip, drop a dot of ink on the center of the horizontal pencil line.  Let this dry a moment & then add more ink to the dot.
  5. Add a small amount of water to the bottom of the chromatography chamber. (The ink dot should be ABOVE the surface of the water.)
  6. Straighten a paper clip and poke a hole through the top of your chromatography strip
  7. Use the paper clip to hang the strip in your chamber. (The straighten paper clip will lay across the top of the chamber.)
  8. MAKE SURE THE TIP OF THE STRIP BUT NOT THE INK IS IMMERSED IN THE WATER!
  9. Notice the separation of the ink as both the solvent and ink travel up the chromatography strip.
  10. Once the solvent front has neared the top of the strip, remove the strip from the chamber and lay it on a piece of paper towel.
  11. Immediately mark the solvent front with a faint pencil line.
  12. Immediately mark the leading edge of each color with an “x”.
  13. Measure, in millimeters, the distance the solvent migrated from the tip of the strip to your solvent front pencil line.
  14. Measure, in millimeters, the distance each color migrated from the point of origin (pencil line where the ink dot was placed) to the leading edge of the color (marked with an “x”.
  15. Record all data in Data table 1.
  16. Calculate and record the Rf value for each color using the formula below.

Rf = distance traveled by the compound / distance traveled by the solvent

Data Table 1

 

Color pen/marker used:

Separated colors
(list top of strip to bottom)
Distance each color traveled

(mm)

Distance solvent (H2O)
(mm)
Rf Value for each color

(Distance color traveled / Distance solvent traveled)

       
       
       
       
       
       
       
       

 

 

 

Color pen/marker used:

Separated colors
(list top of strip to bottom)
Distance each color traveled

(mm)

Distance solvent (H2O)
(mm)
Rf Value for each color

(Distance color traveled / Distance solvent traveled)

       
       
       
       
       
       
       
       

 

 

Questions:

1. Which color of marker did you use?

2. which color separated out first from your ink dot?

3. Why did the inks separate?

 

4. What was your solvent?

5. If you had used markers that weren’t water-soluble, how would you have had to change this lab?

 

6. Why did some inks move a greater distance than others?

 

7. How do scientists use paper chromatography in their investigations?

 

 

Homeostasis & Transport

 

HOMEOSTASIS AND TRANSPORT
All Materials © Cmassengale

 

I. Cell Membranes

 

A. Cell membranes help organisms maintain homeostasis by controlling what substances may enter or leave cells

B. Some substances can cross the cell membrane without any input of energy by the cell

C. The movement of such substances across the membrane is known as passive transport

 

D. To stay alive, a cell must exchange materials such as food, water, & wastes with its environment

E. These materials must cross the cell or plasma membrane

F. Small molecules like water, oxygen, & carbon dioxide can move in and out freely

G. Large molecules like proteins & carbohydrates cannot move easily across the plasma membrane

H. The Cell Membrane is semipermeable or selectively permeable only allowing certain molecules to pass through

 

II. Diffusion

 

A. Diffusion is the movement of molecules from an area of higher concentration to an area of lower concentration

B. Small molecules can pass through the cell membrane by a process called diffusion

 

C. Diffusion across a membrane is a type of passive transport because it does not require energy

D. This difference in the concentration of molecules across a membrane is called a concentration gradient

 

E. Diffusion is driven by the kinetic energy of the molecules

F. Kinetic energy keeps molecules in constant motion causing the molecules to move randomly away from each other in a liquid or a gas

G. The rate of diffusion depends on temperature, size of the molecules, & type of molecules diffusing

 

H. Molecules diffuse faster at higher temperatures than at lower temperatures

I. Smaller molecules diffuse faster than larger molecules

J. Most short-distance transport of materials into & out of cells occurs by diffusion

K. Solutions have two parts — the solute which is being dissolved in the solvent

 

L. Water serves as the main solvent in living things

M. Diffusion always occurs down a concentration gradient (water moves from an area where it is more concentrated to an area where it is less concentrated)

N. Diffusion continues until the concentration of the molecules is the same on both sides of a membrane

 

O. When a concentration gradient no longer exists, equilibrium has been reached but molecules will continue to move equally back & forth across a membrane

 

III. Osmosis

 

A. The diffusion of water across a semipermeable membrane is called osmosis

B. Diffusion occurs from an area of high water concentration (less solute) to an area of lower water concentration (more solute)

 

C. Movement of water is down its concentration gradient & doesn’t require extra energy

D. Cytoplasm is mostly water containing dissolved solutes

E. Concentrated solutions have many solute molecules & fewer water molecules

F. Water moves from areas of low solute concentration to areas of high solute concentration

G. Water molecules will cross membranes until the concentrations of water & solutes is equal on both sides of the membrane; called equilibrium

 

H. At equilibrium, molecules continue to move across membranes evenly so there is no net movement

I. Hypertonic Solution
1. Solute concentration outside the cell is higher (less water)
2. Water diffuses out of the cell until equilibrium is reached
3. Cells will shrink & die if too much water is lost
4. Plant cells become flaccid (wilt); called plasmolysis

J. Hypotonic Solution
1. Solute concentration greater
inside the cell (less water)
2. Water moves into the cell until equilibrium is reached
3. Animal cells swell & burst (lyse) if they take in too much water
4. Cytolysis is the bursting of cells
5. Plant cells become turgid due to water pressing outward against cell wall
6. Turgor pressure in plant cells helps them keep their shape
7. Plant cells do best in hypotonic solutions

K. Isotonic Solutions
1. Concentration of solutes same inside & outside the cell
2. Water moves into & out of cell at an equal rate so there is no net movement of water
3. Animal cells do best in isotonic solutions

 

IV. How Cells Deal With Osmosis

 

A. The cells of animals on land are usually in isotonic environment (equilibrium)

B. Freshwater organisms live in hypotonic environments so water constantly moves into their cells

C. Unicellular freshwater organisms use energy to pump out excess water by contractile vacuoles

D. Plant cell walls prevent plant cells from bursting in hypotonic environments

E. Some marine organisms can pump out excess salt

 

V. Facilitated Diffusion

 

A. Faster than simple diffusion

B. Considered passive transport because extra energy not used

C. Occurs down a concentration gradient

D. Involves carrier proteins embedded in a cell’s membrane to help move across certain solutes such as glucose

 

E. Carrier molecules change shape when solute attaches to them

F. Change in carrier protein shape helps move solute across the membrane

G. Channel proteins in the cell membrane form tunnels across the membrane to move materials

H. Channel proteins may always be open or have gates that open & close to control the movement of materials; called gated channels

 

I. Gates open & close in response to concentration inside & outside the cell

 

VI. Active Transport

 

A. Requires the use of ATP or energy

B. Moves materials against their concentration gradient from an area of lower to higher concentration

C. May also involve membrane proteins

D. Used to move ions such as Na+, Ca+, and K+ across the cell membrane

E. Sodium-Potassium pump moves 3 Na+ out for every 2 K+ into the cell
1. Causes a difference in charge inside and outside the cell
2. Difference in charge is called membrane potential

 

F. Ion pumps help muscle & nerve cells work

 

G. Plants use active transport to help roots absorb nutrients from the soil (plant nutrients are more concentrated inside the root than outside)

 

VII. Bulk Transport

 

A. Moves large, complex molecules such as proteins across the cell membrane

B. Large molecules, food, or fluid droplets are packaged in membrane-bound sacs called vesicles

 

C. Endocytosis moves large particles into a cell

D. Phagocytosis is one type of endocytosis
1. Cell membrane extends out forming pseudopods (fingerlike projections) that surround the particle
2. Membrane pouch encloses the material & pinches off inside the cell making a vesicle
3. Vesicle can fuse with lysosomes (digestive organelles) or release their contents in the cytoplasm
4. Used by ameba to feed & white blood cells to kill bacteria
5. Known as “cell eating”

 

E. Pinocytosis is another type of endocytosis
1. Cell membrane surrounds fluid droplets
2. Fluids taken into membrane-bound vesicle
3. Known as “cell drinking”

 

F. Exocytosis is used to remove large products from the cell such as wastes, mucus, & cell products

G. Proteins made by ribosomes in a cell are packaged into transport vesicles by the Golgi Apparatus

H. Transport vesicles fuse with the cell membrane and then the proteins are secreted out of the cell (e.g. insulin)

BACK

Genetic Traits Activity

 

Finding Your Genetic Match

Introduction:

Have you ever noticed that brothers or sisters often look alike?  Their inherited traits are what make their physical appearance so similar. An inherited trait is a particular genetically determined characteristic that distinguishes a person. The traits of children are determined by the traits that  are passed on from their parents. Some traits are obvious in a family — a child’s nose is shaped like their mother’s nose, but some traits are less obvious. You may have similar traits to many of your classmates even though you are not related to them. Some examples of often un-noticed human traits are the ability or not to roll your tongue, attached or unattached earlobes, dimples or freckles, naturally curly or straight hair, hitchhiker’s or straight thumb, straight or widow’s peak hairline, smooth or cleft chin, or colorblindness or normal vision.

There are numerous traits in humans, but some traits occur more frequently than others.  Between 70-90% of the human population have free-hanging earlobes, can roll their tongue,  are right-handed, and can taste a chemical called PTC.  These traits are called high frequency traits.

Objective:

Students will determine the presence of certain high frequency traits in themselves & their classmates.

Materials:

Genetic Inventory sheet with pictures, paper, pencil, PTC taste strips.

Procedure:

  1. Identify which of the following 10 human traits you have by placing a check mark beside that trait.
  2. Compare the traits you have with other students in the classroom and find the student you most closely match.

 

 

Human Trait Inventory
Student:
Tongue Roller
Non-Tongue Roller
Attached Earlobes
Unattached earlobes
Dimples
No Dimples
Right-handed
Left-Handed
Widow’s Peak
Straight Hairline
Left Thumb on top when Hands Crossed
Right Thumb on top when Hands Crossed
Hair on mid-digit of hand
No hair on mid-digit of hand
Bent little finger
Straight little finger
Second toe longer than big toe
Second toe not longer than big toe
Can Taste PTC
Can Not Taste PTC
Vulcan (Fingers spread 2 by 2)
None Vulcan
Class Match:

 

 

 

Tongue Roller Non Roller Dimples No Dimples
Attached Earlobes Unattached Earlobes Widow’s Peak Straight Hairline
Longer Second Toe Short Second Toe Bent Little finger Hitchhiker’s Thumb
Attached Ear lobes (left)
Unattached ear Lobes (right)
“VULCAN” or No “VULCAN” Dimples Right/Left Thumb on top

 

Genetics PPT Questions

 

 

Mendelian Genetics
PowerPoint Questions
Gregor Mendel

1. Who is responsible for our laws of inheritance?

2. What organism did Mendel study?

3. When was Mendel’s work recognized?

4. When did Mendel perform his experiments & how many plants did he grow?

5. What did Mendel notice about offspring traits?

6. How is Mendel referred to today?

7. In what country did Mendel do his research on peas?

8. Mendel stated that physical traits were inherited as _______________.

9. Today we know that particles are actually what?

Terminology

10. Define these three terms:
a. trait –

 

b. heredity –

c. genetics –

 

11. Name & describe two types of genetic crosses.

 

 

12. What is used to solve genetic crosses?

13. Sketch a Punnett square & show how they are  used to solve a genetics problems.

 

 

 

14. Use a Punnett square to solve a cross between two parents that both have the genotype Yy.

 

 

 

 

15. What are alleles & what are the two forms?

 

16. Explain the difference between dominant & recessive alleles.

 

 

17. Using a letter of the alphabet, show how each allele would be represented.

 

18. What is a genotype and write 3 possible genotypes?

 

19. What is a phenotype and write possible phenotypes for your genotypes in question 18?

 

20. Using these alleles, R = red flower and r = yellow flowers, write all possible genotypes & phenotypes.

 

21. What are homozygous genotypes?

 

22. Write a homozygous dominant genotype.

23. Write a homozygous recessive genotype.

24. What is meant by a heterozygous genotype?

 

25. Write a heterozygous genotype.

26. Heterozygous  genotypes are also called _____________.

27. What two things actually determine an organism’s characteristics?

Pea Experiments

28. Give 4 reasons that Mendel used garden peas, Pisum sativum, for his experiments.

 

 

 

29. Name the male and female parts of a flowering plant and explain how pollination occurs.

 

 

30. What is the difference between self and cross pollination?

 

31. Explain how Mendel cross pollinated his pea plants.

 

 

32. How did Mendel get pure plants?

33. Name 8 pea plant traits and give the dominant & recessive form of each.

 

 

 

 

 

34. How did Mendel’s experimental results compare to the theoretical genotypic ratios? Explain.

 

35. What does P1 mean?

36. What is the F1 generation?

37. What is the F2 generation?

38. What results from this cross — TT  x  tt?

39. What results do you get from crossing two hybrids (Tt   x  Tt)?

 

40. Show all your work for solving a P1 monohybrid cross for seed shape.
Trait:
Alleles:

P1 cross:  __________ x __________

Genotype ____________
Phenotype ___________
G. Ratio _____________
P. Ratio _____________

 

41. The offspring of the above cross are called the _____ generation.

42. Show all your work for solving a F1 monohybrid cross for seed shape.
Trait:
Alleles:

F1 cross:  __________ x __________

Genotype ____________
Phenotype ___________
G. Ratio _____________
P. Ratio _____________

43. Show all your work for solving both F2 monohybrid crosses for seed shape.

Trait:
Alleles:

F2 cross:  ________ x ________  F2 cross:  ________ x ________

 

 

 

 

Genotype ____________                  Genotype ____________
Phenotype ___________                   Phenotype ___________
G. Ratio _____________                   G. Ratio _____________
P. Ratio _____________                    P. Ratio _____________

Mendel’s Laws

Complete the following question:

44. _________ are responsible for inherited traits.

45. Phenotype is based on _______________.

46. Each trait requires _____ genes, one from each ____________.

47. State the Law of Dominance and give an example.

 

 

48. State the Law of Segregation and tell when alleles are “recombined”.

 

 

49. State the Law of Independent assortment & tell what type of crosses show this.

 

 

50. Using the formula 2n where n = the number of heterozygotes, tell how many gametes will be produced by each of the following allele combinations:
a. RrYy
b. AaBbCCDd
c. MmNnOoPPQQRrssTtQq

51. What are the possible allele combinations in the egg and sperm from the following cross — RrYy x RrYy.

 

52. Show how to work an F1 dihybrid cross for seed shape & seed color.

Traits:
Alleles:

 

 

F1 cross   __________ x __________

 

 

 

GR         Genotypes           PR         Phenotypes

 

 

 

 

 

 

 

 

53. Complete this cross or crosses for eye color & curliness of the hair — bbC__ x bbcc.

 

 

 

 

54. Draw a table summarizing Mendel’s 3 laws.

 

 

 

 

 

 

Incomplete and Co-Dominance

55. Incomplete dominance occurs in __________ and produces a phenotype _______________ the phenotype of the two parents.

56. Show your work solving a cross for flower color in snapdragons when there is incomplete dominance.

Trait:
Alleles:

Cross:  RR x rr

 

Genotype ____________
Phenotype ___________
G. Ratio _____________
P. Ratio _____________

57. What is codominance & give an example?

 

58. Write the genotypes for each of these blood types:

type A
type B
type AB
type O

59. Solve this codominance problem: IBIB x IAi.

 

 

 

60. Solve this codominance problem for blood type: ii x IAIB.

 

 

 

Sex-Linked Traits

61. What are sex linked traits?

 

62. Name the sex chromosomes.

63. Write the genotype for male and for female.

64. Most sex-linked traits are carried on what chromosome?

65. Give an example of a sex-linked trait in fruit flies.

66. Show the results of crossing a red-eyed male (XRY)  with a white-eyed female (XrXr) fruit fly.
RR =
Rr =
rr =
XY =
XX =

Cross:    __________ x __________

 

 

 

Genotype ____________
Phenotype ___________
G. Ratio _____________
P. Ratio _____________

67. What is meant by a female carrier?

 

68. Name a disease that can be carried in this manner.