- describe and carry out the Benedict’s test for reducing sugars, the iodine test for starch, the emulsion test for lipids and the biuret test for proteins
- describe and carry out a semi-quantitative Benedict’s test on a reducing sugar solution by standardising the test and using the results (time to first colour change or comparison to colour standards) to estimate the concentration
- describe and carry out a test to identify the presence of non-reducing sugars, using acid hydrolysis and Benedict’s solution
Biological molecules
A-Level Biology · Topic 2
2.1
Testing for biological molecules
Syllabus
Source: Cambridge International syllabus
Living things are built from four main kinds of large molecule 分子: carbohydrates 碳水化合物, lipids 脂质, proteins 蛋白质 and nucleic acids 核酸. You can use simple chemical tests to find out which kinds are present in a sample.
Benedict's test: blue turns green, then orange, then brick-red as more reducing sugar is present
| Test | What it finds | Method | Positive result |
|---|---|---|---|
| Benedict's test | reducing sugar 还原糖 | add Benedict's solution and heat in a water bath | blue changes to green, yellow, orange, then brick-red precipitate 沉淀 |
| iodine test | starch 淀粉 | add orange-brown iodine 碘 solution | colour changes to blue-black |
| emulsion test | lipid | mix sample with ethanol, then pour into water | a white, cloudy emulsion 乳浊液 forms |
| biuret 双缩脲 test | protein | add biuret solution at room temperature | blue changes to purple |
Semi-quantitative Benedict's test
The normal Benedict's test only tells you "yes or no". A semi-quantitative 半定量 test gives a rough amount. First you standardise 标准化 the test: you run it on solutions of known concentration 浓度 and record the result for each. Then you can estimate an unknown by either:
- the time to the first colour change (more sugar changes colour faster), or
- comparing the final colour to your set of colour standards.
Testing for non-reducing sugars
Some sugars, such as sucrose, are non-reducing sugar 非还原糖: they give a negative Benedict's test. To detect them:
- Do a normal Benedict's test first. It stays blue (no reducing sugar).
- Take a fresh sample and add dilute hydrochloric acid 盐酸, then heat. This acid hydrolysis breaks the sugar into smaller reducing sugars.
- Cool, then neutralise 中和 the acid with sodium hydrogencarbonate.
- Now do the Benedict's test again. A brick-red colour shows a non-reducing sugar was present.
Testing for a non-reducing sugar: hydrolyse it with acid first, then the second Benedict's test turns red
Worked example. A solution gives a negative Benedict's test. It is then boiled with dilute hydrochloric acid, neutralised with sodium hydrogencarbonate, and re-tested with Benedict's - now it turns brick-red. What was present, and why is each step needed? The first negative result rules out a reducing sugar. Boiling with acid hydrolyses the glycosidic bond, splitting a non-reducing sugar such as sucrose into its reducing monosaccharides. The neutralising step is essential because Benedict's only works in alkaline conditions - skip it and the test fails even when sugar is present. The positive re-test therefore shows a non-reducing sugar was there all along. Quote the first, negative test as part of the answer: without it, the final red colour cannot tell a non-reducing sugar from a reducing one.
Testing for a non-reducing sugar
Step through the trick: a non-reducing sugar stays blue, so you hydrolyse it with acid, neutralise, then re-test.
| English | Chinese | Pinyin |
|---|---|---|
| molecule | 分子 | fèn zǐ |
| carbohydrate | 碳水化合物 | tàn shuǐ huà hé wù |
| lipid | 脂质 | zhī zhì |
| protein | 蛋白质 | dàn bái zhì |
| nucleic acid | 核酸 | hé suān |
| reducing sugar | 还原糖 | huán yuán táng |
| precipitate | 沉淀 | chén diàn |
| starch | 淀粉 | diàn fěn |
| iodine | 碘 | diǎn |
| emulsion | 乳浊液 | rǔ zhuó yè |
| biuret | 双缩脲 | shuāng suō niào |
| semi-quantitative | 半定量 | bàn dìng liàng |
| standardise | 标准化 | biāo zhǔn huà |
| concentration | 浓度 | nóng dù |
| non-reducing sugar | 非还原糖 | fēi huán yuán táng |
| hydrochloric acid | 盐酸 | yán suān |
| neutralise | 中和 | zhōng hé |
2.2
Carbohydrates
Syllabus
- describe and draw the ring forms of α-glucose and β-glucose
- define the terms monomer, polymer, macromolecule, monosaccharide, disaccharide and polysaccharide
- state the role of covalent bonds in joining smaller molecules together to form polymers
- state that glucose, fructose and maltose are reducing sugars and that sucrose is a non-reducing sugar
- describe the formation of a glycosidic bond by condensation, with reference to disaccharides, including sucrose, and polysaccharides
- describe the breakage of a glycosidic bond in polysaccharides and disaccharides by hydrolysis, with reference to the non-reducing sugar test
- describe the molecular structure of the polysaccharides starch (amylose and amylopectin) and glycogen and relate their structures to their functions in living organisms
- describe the molecular structure of the polysaccharide cellulose and outline how the arrangement of cellulose molecules contributes to the function of plant cell walls
- state that triglycerides are non-polar hydrophobic molecules and describe the molecular structure of triglycerides with reference to fatty acids (saturated and unsaturated), glycerol and the formation of ester bonds
- relate the molecular structure of triglycerides to their functions in living organisms
- describe the molecular structure of phospholipids with reference to their hydrophilic (polar) phosphate heads and hydrophobic (non-polar) fatty acid tails
Source: Cambridge International syllabus
Monomers, polymers and macromolecules
- a monomer 单体 is a small molecule that is a single unit.
- a polymer 聚合物 is a long molecule made of many monomers joined together.
- a macromolecule 大分子 is any very large molecule.
Sugars come in three sizes:
- a monosaccharide 单糖 is a single sugar unit, such as glucose 葡萄糖 and fructose.
- a disaccharide 二糖 is two units joined, such as maltose and sucrose.
- a polysaccharide 多糖 is many units joined into a polymer.
Glucose, fructose 果糖 and maltose 麦芽糖 are reducing sugars. Sucrose 蔗糖 is a non-reducing sugar.
Two ring forms of glucose
Glucose has six carbon atoms and forms a ring. There are two ring forms. They differ only at carbon 1:
- in α-glucose, the –OH group on carbon 1 points down, below the ring.
- in β-glucose, the –OH group on carbon 1 points up, above the ring.
This small difference decides which polysaccharide the glucose can build.
The two ring forms differ only at carbon 1: the –OH points down in α, up in β
Joining and breaking sugars
Monomers are joined by strong covalent bonds 共价键. When two sugars join, a glycosidic bond 糖苷键 forms between them. This happens by condensation 缩合: a molecule of water is removed each time a bond forms.
The reverse is hydrolysis 水解: a water molecule is added to break a glycosidic bond. This is why the non-reducing sugar test needs acid and heat — they hydrolyse sucrose into glucose and fructose.
Condensation removes water to join monomers; hydrolysis adds water to split them
Storage polysaccharides: starch and glycogen
Starch is the energy 能量 store in plants. It is made of two polymers of α-glucose:
- amylose 直链淀粉 — a long, unbranched chain that coils into a spiral.
- amylopectin 支链淀粉 — a chain with many side branches.
Glycogen 糖原 is the energy store in animals. It is like amylopectin but has even more branches, so it can be broken down quickly when energy is needed.
These stores suit their job well: they are compact, they are insoluble 不溶 (so they do not leave the cell), and they do not change the water potential 水势 of the cell (so they do not pull water in by osmosis 渗透). The many branches give many ends, so glucose can be added or removed fast.
Real starch grains inside a potato, stained by iodine. Each grain is a dense, insoluble package of amylose and amylopectin — note the scale: the largest are only about 0.1 mm across
Cellulose
Cellulose 纤维素 is made of β-glucose. Because of the β form, every other glucose is flipped over, so the chains are long and straight. Many straight chains lie side by side and are held together by hydrogen bonds 氢键 into strong bundles called microfibrils 微纤丝. These give the plant cell wall 细胞壁 its strength and stop the cell bursting.
The shape fits the job: amylose coils and amylopectin/glycogen branch (for compact stores), while straight cellulose chains pack into strong fibres
Condensation and hydrolysis
Step through how two sugars join. Condensation removes one water to make the bond; hydrolysis is the reverse — adding water splits it again.
| English | Chinese | Pinyin |
|---|---|---|
| monomer | 单体 | dān tǐ |
| polymer | 聚合物 | jù hé wù |
| macromolecule | 大分子 | dà fēn zi |
| monosaccharide | 单糖 | dān táng |
| glucose | 葡萄糖 | pú táo táng |
| disaccharide | 二糖 | èr táng |
| polysaccharide | 多糖 | duō táng |
| fructose | 果糖 | guǒ táng |
| maltose | 麦芽糖 | mài yá táng |
| sucrose | 蔗糖 | zhè táng |
| covalent bond | 共价键 | gòng jià jiàn |
| glycosidic bond | 糖苷键 | táng gān jiàn |
| condensation | 缩合 | suō hé |
| hydrolysis | 水解 | shuǐ jiě |
| energy | 能量 | néng liàng |
| amylose | 直链淀粉 | zhí liàn diàn fěn |
| amylopectin | 支链淀粉 | zhī liàn diàn fěn |
| glycogen | 糖原 | táng yuán |
| insoluble | 不溶 | bù róng |
| water potential | 水势 | shuǐ shì |
| osmosis | 渗透 | shèn tòu |
| cellulose | 纤维素 | xiān wéi sù |
| hydrogen bond | 氢键 | qīng jiàn |
| microfibril | 微纤丝 | wēi xiān sī |
| cell wall | 细胞壁 | xì bāo bì |
2.2
Lipids
Triglycerides
A triglyceride 甘油三酯 is the main fat or oil. It is non-polar 非极性 and hydrophobic 疏水 (it does not mix with water). It is made from one glycerol 甘油 molecule joined to three fatty acids 脂肪酸 by ester bonds 酯键. Each ester bond forms by condensation, so three water molecules are removed.
One glycerol plus three fatty acid tails; a straight tail is saturated, a kinked one unsaturated
Fatty acids are of two kinds:
- saturated 饱和 — no carbon–carbon double bonds 双键; these fats are usually solid.
- unsaturated 不饱和 — one or more double bonds; these oils are usually liquid.
Triglycerides make a good long-term energy store: they release about twice as much energy per gram as carbohydrates, they are insoluble, and they store little extra mass because they hold no water. Under the skin they also give insulation 隔热 and protect the organs.
Phospholipids
A phospholipid 磷脂 is like a triglyceride, but one fatty acid is replaced by a phosphate 磷酸 group. This gives the molecule two ends with different behaviour:
- a hydrophilic 亲水 ("water-loving") polar 极性 phosphate head.
- two hydrophobic ("water-fearing") fatty acid tails.
This split personality is why phospholipids form the membranes around cells.
The hydrophilic heads face the water; the hydrophobic tails hide inside, forming a bilayer
Building a triglyceride
Watch one glycerol join three fatty acids. Each ester bond forms by condensation, removing one water — three bonds, three waters.
| English | Chinese | Pinyin |
|---|---|---|
| triglyceride | 甘油三酯 | gān yóu sān zhǐ |
| non-polar | 非极性 | fēi jí xìng |
| hydrophobic | 疏水 | shū shuǐ |
| glycerol | 甘油 | gān yóu |
| fatty acid | 脂肪酸 | zhī fáng suān |
| ester bond | 酯键 | zhǐ jiàn |
| saturated | 饱和 | bǎo hé |
| double bond | 双键 | shuāng jiàn |
| unsaturated | 不饱和 | bù bǎo hé |
| insulation | 隔热 | gé rè |
| phospholipid | 磷脂 | lín zhī |
| phosphate | 磷酸 | lín suān |
| hydrophilic | 亲水 | qīn shuǐ |
| polar | 极性 | jí xìng |
2.3
Proteins
Syllabus
- describe and draw the general structure of an amino acid and the formation and breakage of a peptide bond
- explain the meaning of the terms primary structure, secondary structure, tertiary structure and quaternary structure of proteins
- describe the types of interaction that hold protein molecules in shape: • hydrophobic interactions • hydrogen bonding • ionic bonding • covalent bonding, including disulfide bonds
- state that globular proteins are generally soluble and have physiological roles and fibrous proteins are generally insoluble and have structural roles
- describe the structure of a molecule of haemoglobin as an example of a globular protein, including the formation of its quaternary structure from two alpha (α) chains (α–globin), two beta (β) chains (β–globin) and a haem group
- relate the structure of haemoglobin to its function, including the importance of iron in the haem group
- describe the structure of a molecule of collagen as an example of a fibrous protein, and the arrangement of collagen molecules to form collagen fibres
- relate the structures of collagen molecules and collagen fibres to their function
Source: Cambridge International syllabus
Amino acids and the peptide bond
Proteins are polymers of amino acids 氨基酸. Every amino acid has the same general structure around a central carbon atom: an amino group 氨基 (–NH₂), a carboxyl group 羧基 (–COOH), a hydrogen atom, and a variable side chain 侧链 (the R group). The R group is different in each amino acid.
Two amino acids join by condensation. The bond formed between the amino group of one and the carboxyl group of the next is a peptide bond 肽键, and a water molecule is removed. Many amino acids joined this way make a polypeptide 多肽. Adding water (hydrolysis) breaks a peptide bond.
Every amino acid has an amino group, a carboxyl group and an R group; two join by a peptide bond
Four levels of protein structure
| Level | What it means |
|---|---|
| primary structure 一级结构 | the order of amino acids in the chain |
| secondary structure 二级结构 | local shapes — the α-helix 螺旋 and the β-pleated sheet 折叠片 — held by hydrogen bonds |
| tertiary structure 三级结构 | the whole chain folded into a precise 3-D shape |
| quaternary structure 四级结构 | two or more polypeptide chains joined into one protein |
The four levels: primary → secondary → tertiary → quaternary structure
The folded shape is held together by four kinds of interaction between R groups:
- hydrophobic interactions 疏水作用 (non-polar R groups cluster away from water).
- hydrogen bonding.
- ionic bonds 离子键 (between charged R groups).
- covalent bonding, including strong disulfide bonds 二硫键.
Globular and fibrous proteins
- globular proteins 球状蛋白质 fold into a rounded shape, are usually soluble 可溶, and do jobs in the body (for example enzymes and haemoglobin).
- fibrous proteins 纤维状蛋白质 form long strands, are usually insoluble, and give structure and support (for example collagen).
Haemoglobin — a globular protein
Haemoglobin 血红蛋白 carries oxygen 氧气 in red blood cells. It has a quaternary structure made of four polypeptide chains: two alpha (α-globin) chains and two beta (β-globin) chains. Each chain holds a haem group 血红素. At the centre of each haem group is an iron 铁 atom, and this is where one oxygen molecule binds. Four chains mean one haemoglobin molecule can carry four oxygen molecules.
A real haemoglobin molecule, worked out from X-ray data. Count them: two alpha chains (red), two beta chains (blue), and one green haem group held in each — so four oxygen molecules in total
Collagen — a fibrous protein
Collagen 胶原蛋白 gives strength to skin, tendons 肌腱, bone and blood vessel walls. One collagen molecule is three polypeptide chains wound tightly around each other in a triple strand, held by hydrogen bonds. Many of these molecules lie side by side, slightly staggered, and are cross-linked into thick fibres 纤维. The staggered, cross-linked arrangement makes collagen very strong when pulled.
The four levels of protein structure
Build a protein up one level at a time: sequence → local shapes → a folded 3-D shape → several chains joined.
| English | Chinese | Pinyin |
|---|---|---|
| amino acid | 氨基酸 | ān jī suān |
| amino group | 氨基 | ān jī |
| carboxyl group | 羧基 | suō jī |
| side chain | 侧链 | cè liàn |
| peptide bond | 肽键 | tài jiàn |
| polypeptide | 多肽 | duō tài |
| primary structure | 一级结构 | yī jí jié gòu |
| secondary structure | 二级结构 | èr jí jié gòu |
| helix | 螺旋 | luó xuán |
| pleated sheet | 折叠片 | zhé dié piàn |
| tertiary structure | 三级结构 | sān jí jié gòu |
| quaternary structure | 四级结构 | sì jí jié gòu |
| hydrophobic interactions | 疏水作用 | shū shuǐ zuò yòng |
| ionic bond | 离子键 | lí zi jiàn |
| disulfide bond | 二硫键 | èr liú jiàn |
| globular protein | 球状蛋白质 | qiú zhuàng dàn bái zhì |
| soluble | 可溶 | kě róng |
| fibrous protein | 纤维状蛋白质 | xiān wéi zhuàng dàn bái zhì |
| haemoglobin | 血红蛋白 | xuè hóng dàn bái |
| oxygen | 氧气 | yǎng qì |
| haem group | 血红素 | xuè hóng sù |
| iron | 铁 | tiě |
| collagen | 胶原蛋白 | jiāo yuán dàn bái |
| tendon | 肌腱 | jī jiàn |
| fibre | 纤维 | xiān wéi |
2.4
Water
Syllabus
- explain how hydrogen bonding occurs between water molecules and relate the properties of water to its roles in living organisms, limited to solvent action, high specific heat capacity and latent heat of vaporisation
Source: Cambridge International syllabus
Water is a small molecule, but its two O–H bonds are polar: the oxygen end is slightly negative and the hydrogen ends are slightly positive. So one water molecule attracts its neighbours, forming weak hydrogen bonds between them. These hydrogen bonds explain water's useful properties:
- solvent action — water is a good solvent 溶剂, so many substances dissolve in it. This lets reactions happen and lets substances be carried around the body.
- high specific heat capacity 比热容 — water needs a lot of energy to warm up, so its temperature stays steady. This protects living things from quick temperature changes.
- latent heat of vaporisation 汽化潜热 — water needs a lot of energy to evaporate 蒸发. So when water evaporates (for example as sweat dries), it carries away a lot of heat and cools the body.
Water is polar (δ− oxygen, δ+ hydrogens), so its molecules attract each other by hydrogen bonds — the reason for all the properties above
Why water is polar
Tap each part. Oxygen pulls the shared electrons closer, so it is slightly negative and the hydrogens slightly positive — and these opposite charges form hydrogen bonds.
| English | Chinese | Pinyin |
|---|---|---|
| solvent | 溶剂 | róng jì |
| specific heat capacity | 比热容 | bǐ rè róng |
| latent heat of vaporisation | 汽化潜热 | qì huà qián rè |
| evaporate | 蒸发 | zhēng fā |
2.4
Exam tips
- Learn each food test as reagent + positive result + colour change (Benedict's → brick-red; iodine → blue-black; biuret → purple; emulsion → white).
- Benedict's detects reducing sugars; for a non-reducing sugar you must hydrolyse with acid first, then re-test.
- Name the bond precisely: glycosidic (carbohydrates), ester (lipids), peptide (proteins) — all made by condensation (water removed).
- Link structure to function: cellulose (straight chains, H-bonds → strong), glycogen/starch (branched/coiled → compact store).
- For proteins, name the bond at each level: primary (peptide), secondary (hydrogen), tertiary (R-group interactions), quaternary.