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PERMEABILITY PROPERTIES I: ARTIFICIAL MEMBRANES Cargando, Debei Fajardo, Laiza Ferrer, Liezel Pajarillaga, Shara Ramos, Angeli

Experiment 2 (Group3)

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Page 1: Experiment 2 (Group3)

PERMEABILITY PROPERTIES I:

ARTIFICIAL MEMBRANES Cargando, Debei

Fajardo, Laiza

Ferrer, Liezel

Pajarillaga, Shara

Ramos, Angeli

Page 2: Experiment 2 (Group3)

INTRODUCTION

Page 3: Experiment 2 (Group3)

MEMBRANE PERMEABILITY

Quality of a cell’s plasma membrane that allows substances to pass in and out of the cell

Cell membranes have SELECTIVE PERMEABILITY.

The membrane will allow certain substances to pass while forming a barrier against others.

Dependent upon the bilayer structure of the cell membrane

Page 4: Experiment 2 (Group3)

CELL MEMBRANE

barrier between the inside of the cell (ICF: cytoplasm and organelles) and the outside of the cell (ECF)

there is a constant and dynamic exchange of substances between the two fluids

Page 5: Experiment 2 (Group3)

CELL MEMBRANE

flexible lipid bilayer phospholipid

polar, hydrophilic head 2 hydrophobic hydrocarbon tails spontaneously bury the tails together and leave the hydrophilic

heads exposed automatically fixes itself when torn

3 major classes of lipid molecules: (1) phospholipids, (2) cholesterol, (3) glycolipids diff membranes have diff ratios of the 3 lipids

surface proteins functions: (1) cell surface receptors, (2) enzymes, (3) surface

antigens, (4) transporters also has hydrophilic and hydrophobic regions

hydrophilic region: help anchor the protein inside the cell membrane can be classified as (1) intrinsic / integral or (2) extrinsic / peripheral

Page 6: Experiment 2 (Group3)
Page 7: Experiment 2 (Group3)
Page 8: Experiment 2 (Group3)

IMPORTANCE OF MEMBRANE PERMEABILITY

the cells can extract the amino acids, fatty acids, sugars and vitamins it needs to carry out its daily functions

enables chemical messengers, such as hormones, to enter the cell and either trigger / inhibit some function of the cell as necessary

allows the exit and release of waste products after the cells create hormones, neurotransmitters,

proteins and other substances required by the body, MP allows these substances to exit the cell

prevents harmful substances from infiltrating the cell

ensures that the cell does not lose too much of its precious fluids and proteins at any one time

Page 9: Experiment 2 (Group3)

METHODOLOGY &

GUIDE QUESTIONS

Page 10: Experiment 2 (Group3)

Solution A

0.20 g NaCl

50 ml dH20

Solution B

0.20 g NaCl

50 ml

gelatin soln

Page 11: Experiment 2 (Group3)

WHAT IS THE PURPOSE OF GELATIN?

Page 12: Experiment 2 (Group3)

GELATIN

Comes from the thermal denaturation of collagen, isolated from animal skin and bones, with very dilute acid.

Gelatin is only partially soluble in cold water, however dry gelatin swells or hydrates when stirred into water. On warming to about 40°C gelatin that has been allowed to hydrate for about 30 minutes melts to give a uniform solution.

A protein food composed of 18 amino acids.

Page 13: Experiment 2 (Group3)

GELATIN

What is the purpose of the 5% gelatin solution in solution B?

The 5% gelatin solution simulates the inside of an animal cell. It contains proteins and peptides that are impermeable to the longganisa skin, which acts as the membrane.

Page 14: Experiment 2 (Group3)

Soln

A

Soln

B

Maintain distance

Page 15: Experiment 2 (Group3)
Page 16: Experiment 2 (Group3)

Soln BB

Page 17: Experiment 2 (Group3)

WHY LABEL SOLUTION BB DIFFERENTLY

FROM SOLUTION B?

Page 18: Experiment 2 (Group3)

Solution B → Solution BB

Concentration difference (permeating solute)

results in diffusion from area of high

concentration to low concentration

Solution BB has more Cl- ions because of

diffusion from solution A to B along the

concentration gradient

Page 19: Experiment 2 (Group3)

Soln A

0.5 ml

Soln A

Page 20: Experiment 2 (Group3)

0.5 ml

Soln A +

50 uL

Potassium

Chromate

+ Silver

Nitrate

25 uL

increments

Page 21: Experiment 2 (Group3)

WHY USE SILVER NITRATE?

Page 22: Experiment 2 (Group3)

Silver nitrate can react with the chloride ions of the NaCl in the solution.

Forms a while solid ppt -> AgCl-

Chemical reaction:

NaCl → Na+ + Cl-

AgNO3 + Cl- → AgCl- + NO3-

Page 23: Experiment 2 (Group3)

WHY USE POTASSIUM CHROMATE?

Page 24: Experiment 2 (Group3)

Potassium chromate was used because it indicates that there are no more chloride ions present in the solution. Excess silver nitrate (from the silver nitrate-chloride reaction) reacts

with potassium chromate

Product: silver chromate

(brick red ppt)

K2CrO4 →2K+ + CrO42+

Ag+ + CrO42+ → Ag2CrO4

-

Page 25: Experiment 2 (Group3)

DISCUSSION

Page 26: Experiment 2 (Group3)

Solution Amount of Silver

Nitrate (uL)

Computed moles of

Cl

A 725 ?

BB 250 ?

Conversion factor:

Each 50 µL portion of AgNO3

represent 1 mg of NaCl / mL

General Formula:

Molecula

r weight

Simple

conversion AgNO3 + Cl- AgCl- + NO3

-

Page 27: Experiment 2 (Group3)

COMPUTATIONS

For Soln A:

= 2.48 x 10 -4 mol Cl-

725

Page 28: Experiment 2 (Group3)

COMPUTATIONS

For Soln BB:

= 8.55 x 10 -5 mol Cl-

250

Page 29: Experiment 2 (Group3)

Solution Amount of Silver

Nitrate (uL)

Computed moles of

Cl

A 725 2.48 x 10 -4

BB 250 8.55 x 10 -5

Page 30: Experiment 2 (Group3)

GIBBS-DONNAN RULE

the presence of a nonpermeating charged solute on one side of a membrane results in an equilibrium in which each permeating charged solute is more concentrated on one side than the other

The final distribution of all ions will satisfy 3 requirements:

i. the total number of cations and anions on the same side of the semi-permeable membrane will be equal

# of cations = # of anions protein

# of cations = # of anions

Page 31: Experiment 2 (Group3)

ii. on the side containing the protein, the number of diffusible ANIONS present will be less and the number of diffusible CATIONS will be greater than on the other side of the semi-permeable membrane

# of cations

# of anions

protein

# of cations

# of anions

>

<

iii. the osmotic pressure on the side containing the protein anion will be slightly greater than on the side without protein anions

Page 32: Experiment 2 (Group3)

GIBBS-DONNAN EQUILIBRIUM IN SET-UP

Na+

Protein –

Na+

Cl –

Starting condition

membrane

Sol B Sol A

Initial concentrations (molar) N

a+

Pro

tein

Na+

Cl –

Page 33: Experiment 2 (Group3)

Na+

Protein—

Cl –

Na+

Cl –

Net diffusional movements during approach to equilibrium

membrane

Sol B Sol A

Final concentrations (molar)

Na

+

Na

+

Cl–

Na+

Cl –

Pro

tein

– C

l–

Sol BB Sol A

[Na]1 [Cl]1 = [Na]2 [Cl]2

Page 34: Experiment 2 (Group3)

Final concentrations (molar)

Na

+

Cl–

Sol BB Sol A

Pro

tein

– C

l–

Na

+

Solution Amount of Silver

Nitrate (uL)

Computed moles of

Cl

A 725 2.48 x 10 -4

BB 250 8.55 x 10 -5

Page 35: Experiment 2 (Group3)

HOW DOES THIS PHENOMENON AFFECT

THE GENERATION OF A MEMBRANE

POTENTIAL IN LIVING CELLS?

Aside from generating a concentration gradient, there is also a difference of electrical potential at equilibrium, with the nonpermeating solute having the charge of that solute.

Na+

Protein—

Cl –

Na+

Cl –

membrane

Sol B Sol A

Na+

Cl –

Page 36: Experiment 2 (Group3)

REFERENCES

http://www.biology4kids.com/files/cell_membrane.html

http://library.thinkquest.org/C004535/cell_membranes.html

http://www.biologymad.com/cells/cellmembrane.htm

http://www.wisegeek.com/what-is-membrane-permeability.htm

http://www.lsbu.ac.uk/water/hygel.html

http://www.gelatin.co.za/gltn1.html

http://www.gmap-gelatin.com/about_gelatin_comp.html