Biological molecules Flashcards

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1
Q

Why is water vital to living organisms?

A

Water is a reactant in loads of important chemical reactions, including hydrolysis reactions (see page 22).
2) Water is a solvent, which means some substances dissolve in it. Most biological reactions take place
in solution (e.g. in the cytoplasm of eukaryotic and prokaryotic cells) so water’s pretty essential.
3) Water transports substances. The fact that it’s a liquid and a solvent means it can easily transport
all sorts of materials, like glucose and oxygen, around plants and animals.
4) Water helps with temperature control because it has a high specific heat capacity
and a high latent heat of evaporation (see below).
5) Water is a habitat. The fact that it helps with temperature control, is a solvent and becomes less dense
when it freezes (see next page) means many organisms can survive and reproduce in it.

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2
Q

What structure do water molecules have?

A

A simple structure-A molecule of water (H O) is one atom of oxygen (O)
2
joined to two atoms of hydrogen (H ) by shared electrons.
2
2) Because the shared negative hydrogen electrons are pulled
towards the oxygen atom, the other side of each hydrogen
atom is left with a slight positive charge.
3) The unshared negative electrons on the
oxygen atom give it a slight negative charge.
4) This makes water a polar molecule — it has a partial negative (delta–)
charge on one side and a partial positive (d+) charge on the other
5) The slightly negatively-charged oxygen
atoms attract the slightly positively-charged
hydrogen atoms of other water molecules.
6) This attraction is called hydrogen bonding and
it gives water some of its useful properties.

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3
Q

Why does water have a high specific heat capacity?

A

Hydrogen Bonds Give Water a High Specific Heat Capacity
1) Specific heat capacity is the energy needed to raise the temperature of 1 gram of a substance by 1 °C.
2) The hydrogen bonds between water molecules can absorb a lot of energy.
So water has a high specific heat capacity — it takes a lot of energy to heat it up.
3) This means water doesn’t experience rapid temperature changes, which is one of the properties that makes
it a — the temperature under water is likely to be more stable than it is on land.

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4
Q

Why does water have high latent heat of evaporation?

A

Hydrogen Bonds Also Give Water a High Latent Heat of Evaporation
1) It takes a lot of energy (heat) to break the hydrogen bonds between water molecules.
2) So water has a high latent heat of evaporation — a lot of energy is used up when water evaporates.
3) This is useful for living organisms because it means water’s great for cooling things. This is why some
mammals, like us, sweat when they’re too hot. When sweat evaporates, it cools the surface of the skin.

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5
Q

Why is water very cohesive?

A

Water’s Polarity Makes it Very Cohesive
1) Cohesion is the attraction between molecules of the same type (e.g. two water molecules).
Water molecules are very cohesive (they tend to stick together) because they’re polar.
2) This helps water to flow, making it great for transporting substances. It also helps water to be
transported up plant stems in the transpiration stream

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6
Q

Why is water a good solvent?

A

1) A lot of important substances in biological reactions are ionic (like salt, for example). This means they’re
made from one positively-charged atom or molecule and one negatively-charged atom or molecule
(e.g. salt is made from a positive sodium ion and a negative chloride ion).
2) Because water is polar, the slightly positive end of a water molecule will be attracted to the negative ion,
and the slightly negative end of a water molecule will be attracted to the positive ion.
3) This means the ions will get totally surrounded by water molecules — in other words, they’ll dissolve.
4) Water’s polarity makes it useful as a solvent in living organisms. E.g. in humans, important ions can dissolve in the water in blood and then be transported around the body.

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7
Q

Why is water less dense while solid?

A

Water’s Less Dense When it’s Solid
1) At low temperatures water freezes — it turns from a liquid to a solid.
2) Water molecules are held further apart in ice than they are in liquid water because each
water molecule forms four hydrogen bonds to other water molecules, making a lattice
shape. This makes ice less dense than liquid water — which is why ice floats.
3) This is useful for living organisms because, in cold temperatures, ice forms an insulating layer on top of water —
the water below doesn’t freeze. So organisms that live in water, like fish, don’t freeze and can still move around.

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8
Q

What is a polymer?

A

1) Most carbohydrates are polymers. A polymer is a molecule made up of
many similar, smaller molecules (called monomers) bonded together.

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9
Q

What are the monomers called that make up carbohydrates?

A

monosaccharide

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10
Q

What is glucose?

A

Glucose is a monosaccharide with six carbon atoms — this means it’s a hexose monosaccharide.

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11
Q

What are two forms of glucose?

A

4) There are two forms of glucose — alpha (a) and beta (b). They both have a ring structure:

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12
Q

Whats good about glucose structure?

A

Glucose’s structure is a-glucose b-glucose
related to its function as the main energy source in
animals and plants. Its structure makes it
soluble so it can be easily transported. Its chemical
bonds contain lots of energy

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13
Q

What is Ribose?

A

Ribose is a monosaccharide with five carbon atoms

— this means it’s a pentose monosaccharide.

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14
Q

What are all carbohydrates are made up of?

A

All carbohydrates are made up of the same three chemical elements —
carbon (C), hydrogen (H) and oxygen (O). For every carbon atom in the
carbohydrate there are usually two hydrogen atoms and one oxygen atom.

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15
Q

How are disaccharides and polysacharides formed?

A

Monosaccharides Join Together to Form Disaccharides and Polysaccharides
1) Monosaccharides are joined together by glycosidic bonds.
2) During synthesis, a hydrogen atom on one monosaccharide bonds to a hydroxyl (OH) group on the other,
releasing a molecule of water. This is called a condensation reaction.
3) The reverse of this synthesis reaction is hydrolysis.
A molecule of water reacts with the glycosidic bond, breaking it apart.
4) A disaccharide is formed when two monosaccharides join together:
5) A polysaccharide is formed when more than two monosaccharides join together:

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16
Q

What are the 3 polysacharides?

A

starch
glycogen
cellulose

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17
Q

Structure and function of starch?

A

Cells get energy from glucose. Plants store excess glucose as starch (when a plant
needs more glucose for energy it breaks down starch to release the glucose).
2) Starch is a mixture of two polysaccharides of alpha-glucose — amylose and amylopectin:
• Amylose — a long, unbranched chain of a-glucose. The angles of the glycosidic
bonds give it a coiled structure, almost like a cylinder. This makes it compact,
so it’s really good for storage because you can fit more in to a small space.
a-
• Amylopectin — a long, branched chain of glucose. Its side branches allow the
enzymes that break down the molecule to get at the glycosidic bonds easily.
This means that the glucose can be released quickly.
3) Starch is insoluble in water, so it doesn’t cause water to enter cells by osmosis
(see p. 58) which would make them swell. This makes it good for storage.

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18
Q

Structure and function of glycogen?

A

Glycogen — the main energy storage material in animals
1) Animal cells get energy from glucose too. But animals store excess
glucose as glycogen — another polysaccharide of alpha-glucose.
2) Its structure is very similar to amylopectin, except that it has loads more
side branches coming off it. Loads of branches means that stored glucose
can be released quickly, which is important for energy release in animals.
3) It’s also a very compact molecule, so it’s good for storage.

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19
Q

Structure and function of cellulose?

A

Cellulose — the major component of cell walls in plants
1) Cellulose is made of long, unbranched chains of beta-glucose.
-
2) When beta glucose molecules bond, they form straight cellulose chains.
3) The cellulose chains are linked together by hydrogen bonds to form strong
fibres called microfibrils. The strong fibres mean cellulose provides structural
support for cells (e.g. in plant cell walls).

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20
Q

What are triglycerides?

A

1) Triglycerides are macromolecules — they’re complex molecules with a relatively large molecular mass.
2) Like all lipids, they contain the chemical elements carbon, hydrogen and oxygen.
3) Triglycerides have one molecule of glycerol with three fatty acids attached to it

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21
Q

Structure of triglyceride?

A

Fatty acid molecules have long ‘tails’ made
of hydrocarbons (compounds that contain
only carbon and hydrogen atoms).
The tails are ‘hydrophobic’ (they repel water
molecules). These tails make lipids insoluble
in water. All fatty acids have the same basic
structure, but the hydrocarbon tail varies.

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22
Q

Why do triglycerides contain ester bonds?

A

1) Triglycerides are synthesised by the formation
of an ester bond between each fatty acid
and the glycerol molecule.
2) Each ester bond is formed by a condensation
reaction (in which a water molecule is released).
3) The process in which triglycerides are
synthesised is called esterification.
4) Triglycerides break down when the ester bonds are
broken. Each ester bond is broken in a hydrolysis
reaction (in which a water molecule is used up).

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23
Q

How many kinds of fatty acids are there?

A

2

saturated and unsaturated.

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24
Q

What’s the difference between unsaturated and saturated fatty acids?

A

Hydrocarbon tails

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25
Q

what is the structure of saturated fatty acids?

A

Saturated fatty acids don’t have any double
bonds between their carbon atoms. The fatty
acid is ‘saturated’ with hydrogen.

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26
Q

What is the structure of unsaturated fatty acids?

A

Unsaturated fatty acids have at least one double bond

between carbon atoms, which cause the chain to kink.

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27
Q

Why are phospholipids similar to Triglycerides?

A

1) Phospholipids are also macromolecules. They’re pretty similar
to triglycerides except one of the fatty acid molecules
is replaced by a phosphate group.
2) The phosphate group is hydrophilic (it attracts water
molecules) and the fatty acid tails are hydrophobic.

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28
Q

How are the properties of triglycerides related to their function?

A

In animals and plants, triglycerides are mainly used as energy storage molecules.
Some bacteria (e.g. Mycobacterium tuberculosis) use triglycerides to store both energy and carbon.
Triglycerides are good for storage because:
1) The long hydrocarbon tails of the fatty acids contain lots of chemical energy —
a load of energy is released when they’re broken down. Because of these tails,
lipids contain about twice as much energy per gram as carbohydrates.
2) They’re insoluble, so they don’t cause water to enter the cells by osmosis (see p. 58)
which would make them swell. The triglycerides bundle together as insoluble droplets
in cells because the fatty acid tails are hydrophobic (water-repelling) — the tails
face inwards, shielding themselves from water with their glycerol heads.

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29
Q

How are the properties of phospholipids related to their functions?

A

Phospholipids are found in the cell membranes of all eukaryotes and prokaryotes.
They make up what’s known as the phospholipid bilayer (see p. 50).
Cell membranes control what enters and leaves a cell.
1) Phospholipid heads are hydrophilic and their tails are hydrophobic, so they form
a double layer with their heads facing out towards the water on either side.
2) The centre of the bilayer is hydrophobic, so water-soluble substances can’t
easily pass through it — the membrane acts as a barrier to those substances.

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30
Q

How are the properties of cholesterol related to their functions?

A

Cholesterol is another type of lipid — it has a hydrocarbon ring structure attached
to a hydrocarbon tail. The ring structure has a polar hydroxyl (OH) group attached
to it. In eukaryotic cells, cholesterol molecules help to regulate the fluidity of the
cell membrane by interacting with the phospholipid bilayer.
1) Cholesterol has a small size and flattened shape — this allows cholesterol
to fit in between the phospholipid molecules in the membrane.
2) At higher temperatures, they bind to the hydrophobic tails of the
phospholipids, causing them to pack more closely together.
This helps to make the membrane less fluid and more rigid.
3) At lower temperatures, cholesterol prevents phospholipids from
packing too close together, and so increases membrane fluidity.

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31
Q

What are proteins made from?

A

Long chains of amino acid

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32
Q

what are proteins?

A

polymers

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33
Q

what are amino acids in the Protein?

A

monomers

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34
Q

How is a dipeptide formed?

A

When two amino acids join together

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35
Q

how is a polypeptide formed?

A

When more than two amino acids joined together

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36
Q

what are proteins made up of?

A

one or more polypeptides

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37
Q

why do different amino acids have different variable groups?

A

All amino acids have the same general structure — a carboxyl group (-COOH) and
an amino group (-NH ) attached to a carbon atom. The difference between different
2
amino acids is the variable group (R on diagram) they contain.
2) All amino acids contain the chemical elements carbon, oxygen,
hydrogen and nitrogen. Some also contain sulfur.

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38
Q

What are amino acids joined together by?

A

peptide bonds to form dipeptides on polypeptides

39
Q

what is a condensation reaction?

A

when a molecule of water is released during the reaction

40
Q

what is a hydrolysis reaction?

A

When a reaction adds a molecule of water to break the peptide bond

41
Q

what are the four structures of proteins?

A

Primary secondary tertiary and quaternary

42
Q

what is the primary structure of a protein?

A

Primary Structure — this is the sequence of amino acids in the polypeptide chain.
Different proteins have different sequences of amino acids in their primary structure.
A change in just one amino acid may change the structure of the whole protein.

43
Q

What is the secondary structure of a protein?

A

Secondary Structure — the polypeptide chain doesn’t remain flat and
straight. Hydrogen bonds form between nearby amino acids in the
a
chain. This makes it automatically coil into an alpha ( ) helix or fold
b
into a beta ( ) pleated sheet — this is the secondary structure.

44
Q

What is the tertiary structure of a protein?

A

Tertiary Structure — the coiled or folded chain of amino
acids is often coiled and folded further. More bonds
form between different parts of the polypeptide chain.
For proteins made from a single polypeptide chain,
the tertiary structure forms their final 3D structure.

45
Q

what is the quaternary structure of a protein?

A

Quaternary Structure — some proteins are made of
several different polypeptide chains held together by bonds.
The quaternary structure is the way these polypeptide chains
are assembled together. E.g. haemoglobin is made of four
polypeptide chains, bonded together. For proteins made from
more than one polypeptide chain, the quaternary structure is
the protein’s final 3D structure.

46
Q

What is the primary structure of protein held together by?

A

peptide between the amino acids

47
Q

what is the secondary structure of a protein held by?

A

Hydrogen bonds

48
Q

What is the tertiary structure of a protein held by?

A

ionic bond
disulfide bonds
hydrophobic and hydrophilic interactions
hydrogen bonds

49
Q

what is ionic bonds?

A

Ionic bonds. These are attractions between negatively-charged R groups and positively-charged
R groups on different parts of the molecule.

50
Q

what disulfide bonds?

A

Disulfide bonds. Whenever two molecules of the amino acid cysteine come close together, the
sulfur atom in one cysteine bonds to the sulfur in the other cysteine, forming a disulfide bond.

51
Q

what are hydrophobic and hydrophilic interactions?

A

Hydrophobic and hydrophilic interactions. When hydrophobic
(water-repelling) R groups are close together in the protein, they tend
to clump together. This means that hydrophilic (water‑attracting)
R groups are more likely to be pushed to the outside, which affects
how the protein folds up into its final structure.

52
Q

What are hydrogen bonds?

A

Hydrogen bonds — these weak bonds form between slightly
positively-charged hydrogen atoms in some R groups and slightly
negatively-charged atoms in other R groups on the polypeptide chain.

53
Q

What is the Quaternary structure?

A

Quaternary structure — this tends to be determined by the tertiary structure of the individual polypeptide
chains being bonded together. Because of this, it can be influenced by all the bonds mentioned above.

54
Q

What are globular proteins?

A

Globular Proteins Are Round and Compact
1) In a globular protein, the hydrophilic R groups on the amino acids tend to be
pushed to the outside of the molecule. This is caused by the hydrophobic and
hydrophilic interactions in the protein’s tertiary structure (see previous page).
2) This makes globular proteins soluble, so they’re easily transported in fluids.
3) Globular proteins have a range of functions in living organisms.

55
Q

What is haemoglobin?

A

HAEMOGLOBIN is a globular protein that carries oxygen around the body in
red blood cells (see page 86). It’s known as a conjugated protein — this means
it’s a protein with a non-protein group attached. The non-protein part is called
a prosthetic group. Each of the four polypeptide chains in haemoglobin has a
prosthetic group called haem. A haem group contains iron, which oxygen binds to.

56
Q

What is insulin?

A

INSULIN is a hormone secreted by the pancreas. It helps to regulate the blood glucose level.
Its solubility is important — it means it can be transported in the blood to the tissues where it acts.
An insulin molecule consists of two polypeptide chains, which are held together by disulfide bonds.

57
Q

what is amylase?

A

AMYLASE is an enzyme (see page 42) that catalyses the breakdown of starch in the
digestive system. It is made of a single chain of amino acids. Its secondary structure contains
both alpha-helix and beta-pleated sheet sections. Most enzymes are globular proteins.

58
Q

what are fibrous proteins?

A

Fibrous proteins are insoluble and strong. They’re structural proteins and are fairly unreactive

59
Q

what are 3 fibrous proteins?

A

Collagen keratin elastin

60
Q

What is collagen?

A

Collagen — found in animal connective tissues, such as bone,
skin and muscle. It is a very strong molecule. Minerals can
bind to the protein to increase its rigidity, e.g. in bone.

61
Q

What is keratin?

A

Keratin — found in many of the external structures of animals, such as skin, hair, nails,
feathers and horns. It can either be flexible (as it is in skin) or hard and tough (as it is in nails).

62
Q

What is elastin?

A

• Elastin — found in elastic connective tissue, such as skin, large blood vessels and some ligaments.
It is elastic, so it allows tissues to return to their original shape after they have been stretched.

63
Q

What is calcium’s chemical symbol and role in biological processes?

A

Ca2+
Involved in the transmission of nerve impulses and the release of insulin
from the pancreas. Acts as a cofactor for many enzymes
e.g. those involved in blood clotting. Is important for bone formation.

64
Q

What is sodium chemical symbol and role in biological processes?

A

Na+
Important for generating nerve impulses, for muscle contraction
and for regulating fluid balance in the body.

65
Q

What is pottasiums chemical symbol and role in biological processes?

A

k+
important for generating nerve impulses, for muscle contraction and for
regulating fluid balance in the body. Activates essential enzymes needed
for photosynthesis in plant cells

66
Q

What is hydrogen chemical symbol and role in biological processes?

A

h+
Affects the pH of substances (more H+ ions than OH– ions in a solution
creates an acid). Also important for photosynthesis reactions that occur
in the thylakoid membranes inside chloroplasts

67
Q

What is ammoniums chemical symbol and role in biological processes?

A

NH4+
Absorbed from the soil by plants and is an important source of nitrogen
+
(which is then used to make, e.g. amino acids, nucleic acids).

68
Q

What is nitrates chemical symbol and role in biological processes?

A

NO3-
Absorbed from the soil by plants and is an important source of nitrogen

(which is then used to make, e.g. amino acids, nucleic acids).

69
Q

What is hydrogencarbonates chemical symbol and role in biological processes?

A

HCO3-

Acts as a buffer, which helps to maintain the pH of the blood

70
Q

What is chlorides chemical symbol and role in biological processes?

A

Cl-
Involved in the ‘chloride shift’ which helps to maintain the pH of the
blood during gas exchange (see p. 87). Acts as a cofactor for the enzyme
amylase (see p. 47). Also involved in some nerve impulses.

71
Q

What is phosphate chemical symbol and role in biological processes?

A

PO4 3-
Involved in photosynthesis and respiration reactions. Needed for the
synthesis of many biological molecules, such as nucleotides (including
3–
ATP), phospholipids, and calcium phosphate (which strengthens bones).

72
Q

What is hydroxide chemical symbol and role in biological processes?

A

OH-
Affects the pH of substances (more OH– ions than H+ ions
in a solution creates an alkali).

73
Q

What is an cation?

A

ion with positive charge

74
Q

what is an anion?

A

ion with negative charge

75
Q

What is sugar the general term for?

A

monnosacharides and disacharides?

76
Q

What can sugar is be classified as?

A

reducing non-reducing

77
Q

how do you test for sugar?

A

Benedict’s test

78
Q

how do you test for reducing sugar?

A

1) Reducing sugars include all monosaccharides (e.g. glucose) and some disaccharides (e.g. maltose and lactose).
2) You add Benedict’s reagent (which is blue) to a sample and heat it
in a water bath that’s been brought to the boil.
The colour of the precipitate blue green yellow orange brick red
changes from:
3) If the test’s positive it will form a coloured precipitate (solid particles suspended in the solution).
4) The higher the concentration of reducing sugar, the further the colour change goes — you can use this to
compare the amount of reducing sugar in different solutions. A more accurate way of doing this is to filter
the solution and weigh the precipitate.

79
Q

How do you test non reducing sugars?

A

1) If the result of the reducing sugars test is negative, there could still be a non-reducing sugar present.
To test for non-reducing sugars, like sucrose, first you have to break them down into monosaccharides.
2) You do this by getting a new sample of the test solution, adding dilute hydrochloric acid and carefully
heating it in a water bath that’s been brought to the boil. You then neutralise it with sodium
hydrogencarbonate. Then just carry out the Benedict’s test as you would for a reducing sugar.
3) If the test’s positive it will form a coloured precipitate (as for the reducing sugars test). If the test’s negative
the solution will stay blue, which means it doesn’t contain any sugar (either reducing or non-reducing).

80
Q

How do you test for glucose?

A

Use Test Strips for Glucose
Glucose can also be tested for using test strips coated in a reagent. The strips are dipped in a test solution and change
colour if glucose is present. The colour change can be compared to a chart to give an indication of the concentration
of glucose present. The strips are useful for testing a person’s urine for glucose, which may indicate they have diabetes.

81
Q

how do you test for starch?

A

Use the Iodine Test for Starch
Just add iodine dissolved in potassium iodide solution to the test sample.
• If starch is present, the sample changes from browny-orange to a dark, blue-black colour.
• If there’s no starch, it stays browny-orange.

82
Q

how do you test for protein?

A

Use the Biuret Test for Proteins
There are two stages to this test.
1) The test solution needs to be alkaline, so first you
add a few drops of sodium hydroxide solution.
2) Then you add some copper(II) sulfate solution.
• If protein is present the solution turns purple.
• If there’s no protein, the solution will stay blue.

83
Q

How to test for lipids?

A

Use the Emulsion Test for Lipids
Shake the test substance with ethanol for about a minute,
then pour the solution into water.
• If lipid is present, the solution will turn milky.
• The more lipid there is, the more noticeable the milky colour will be.
• If there’s no lipid, the solution will stay clear.

84
Q

why is colorimetry used?

A

Used to Determine the Concentration of a Glucose Solution

85
Q

what can you use a colorimeter for?

A

1) You can use Benedict’s reagent and a colorimeter to get a quantitative estimate
of how much glucose (or other reducing sugar) there is in a solution.
2) A colorimeter is a device that measures the strength of a
coloured solution by seeing how much light passes through it
3) A colorimeter measures absorbance (the amount of light absorbed by the solution). The more concentrated the colour of the solution, the higher the absorbance is
4) It’s easiest to measure the concentration of the blue Benedict’s solution that’s left after the test (the paler the solution,
the more glucose there was). So, the higher the glucose concentration, the lower the absorbance of the solution.

86
Q

How do u use a colorimeter?

A

Initially you need to make up several glucose solutions of different,
known concentrations. You can do this using a serial dilution technique:
This is how you’d make five serial dilutions with a dilution factor of 2,
starting with an initial glucose concentration of 40 mM…
1) Line up five test tubes in a rack.
2) Add 10 cm3 of the initial 40 mM glucose solution to the first test
tube and 5 cm3 of distilled water to the other four test tubes.
3) Then, using a pipette, draw 5 cm3 of the solution from the first test
tube, add it to the distilled water in the second test tube and mix the
solution thoroughly. You now have 10 cm3 of solution that’s half as
concentrated as the solution in the first test tube (it’s 20 mM).
4) Repeat this process three more times to create solutions of 10 mM, 5 mM and 2.5 mM.

87
Q

How to make a calibration curve?

A

Once you’ve got your glucose solutions, you need to make a calibration curve. Here’s how:
1) Do a Benedict’s test on each solution (plus a negative control of pure water).
Use the same amount of Benedict’s solution in each case.
2) Remove any precipitate — either leave for 24 hours (so that the precipitate settles out) or centrifuge them.
3) Use a colorimeter (with a red filter) to measure the absorbance
1.0
of the Benedict’s solution remaining in each tube.
4) Use the results to make the calibration curve,
showing absorbance against glucose concentration.
Then you can test the unknown solution in the same way as the known
concentrations and use the calibration curve to find its concentration.

88
Q

What is a biosensor?

A

Biosensors Can Detect Chemicals in a Solution
1) A biosensor is a device that uses a biological molecule, such as an enzyme (see page 42) to detect a chemical.
2) The biological molecule produces a signal (e.g. a chemical signal), which is converted to an electrical signal
by a transducer (another part of the biosensor).
3) The electrical signal is then processed and can be used to work out other information.
Example: Glucose Biosensors
1) A glucose biosensor is used to determine the concentration of glucose in a solution.
2) It does this using the enzyme glucose oxidase and electrodes.
3) The enzyme catalyses the oxidation of glucose at the electrodes — this creates a
charge, which is converted into an electrical signal by the electrodes (the transducer).
4) The electrical signal is then processed to work out the initial glucose concentration.

89
Q

why is chromatography used?

A

Chromatography is Good for Separating and Identifying Things
1) Chromatography is used to separate stuff in a mixture —
once it’s separated out, you can often identify the components.
2) For example, chromatography can be used to separate out and identify biological
molecules such as amino acids, carbohydrates, vitamins and nucleic acids.
3) There are quite a few different types of chromatography — you only need to
know about paper chromatography and thin-layer chromatography.

90
Q

how does chromatography work?

A

1) A MOBILE PHASE — where the molecules can move.
• In both paper and thin-layer chromatography the mobile
phase is a liquid solvent, such as ethanol or water.
2) A STATIONARY PHASE — where the molecules can’t move.
• In paper chromatography the stationary phase
is a piece of chromatography paper.
• In thin-layer chromatography the stationary phase
is a thin (0.1-0.3 mm) layer of solid, e.g. silica gel,
on a glass or plastic plate.

91
Q

Why is paper chromatography used?

A

Paper Chromatography is used to Identify Unknown Amino Acids

92
Q

How to do paper chromatography?

A

1) Draw a pencil line near the bottom of a piece of chromatography
paper and put a concentrated spot of the mixture of amino acids
on it. It’s best to carefully roll the paper into a cylinder with the
spot on the outside so it’ll stand up.
2) Add a small amount of prepared solvent (a mixture of butan-1-ol,
glacial ethanoic acid and water is usually used for amino acids)
to a beaker and dip the bottom of the paper into it (not the spot).
This should be done in a fume cupboard. Cover with a lid to
stop the solvent evaporating.3) As the solvent spreads up the paper, the different amino acids
(solutes) move with it, but at different rates, so they separate out.
4) When the solvent’s nearly reached the top, take the paper out and
mark the solvent front with pencil. Then you can leave the paper
to dry out before you analyse it (see below).
5) Amino acids aren’t coloured, which means you won’t be able to see them on the paper. So before you
can analyse them, you have to spray the paper with ninhydrin solution to turn the amino acids purple.
This should also be done in a fume cupboard and gloves should be worn. (Note: you can’t use ninhydrin to
detect all biological molecules, only proteins and amino acids.)
6) You can then use R values to identify the separated molecules:

93
Q

How to calculate rf value?

A

An R value is the ratio of the distance travelled by a solute to the distance travelled by the solvent. You can calculate it using this formula:
R value of amino acid = distance travelled by solute / distance travelled by solvent
When you’re measuring how far a solute has travelled, you measure
from the point of origin to the vertical centre of the spot.
You can work out what was in a mixture by calculating an R value for each solute and looking each R value up in a database, or table, of known values.

94
Q

How to calculate rf value?

A

An R value is the ratio of the distance travelled by a solute to the distance travelled by the solvent. You can calculate it using this formula:
R value of amino acid = distance travelled by solute / distance travelled by solvent
When you’re measuring how far a solute has travelled, you measure
from the point of origin to the vertical centre of the spot.
You can work out what was in a mixture by calculating an R value for each solute and looking each R value up in a database, or table, of known values.