This Study Resource Was: Worksheet D

SLE212 Biochemistry T1 2020 Assessment Worksheet D
Worksheet D
Carbohydrates
1a) Use the image available in the practical 5 folder to produce a figure of the thin layer
chromatogram that could be published in a scientific journal; below you find an example of what
such a figure can look like.
Indicate relevant information, e.g. samples applied, direction of chromatography, the spots, origins,
running front etc. Crop away unnecessary parts of the image. You can use e.g. Power-Point,
Photoshop or Illustrator or similar software for his task. Write a concise figure legend that allows the
reader to understand what they see. The overall presentation of the figure must be neat and tidy!
The colour of ALL labelling (lines, descriptions, arrows etc) of the figure MUST BE BLUE (you will
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receive 0 marks for question 1a in case that you use a different colour). (1.5 marks)
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Explanatory note: The teaching team strictly enforces the above indicated color requirements in order to ensure the
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originality of 2019 worksheet submissions.
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Both your figure and figure legend have to be inserted in this space. Do NOT use more space.
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glucose
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Cellobiose
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cellotriose
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cellotetrose
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origin
DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

DO NOT DELETE ANY TEXT FROM THE QUESTIONS
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1b) Thin Layer chromatography (TLC). i) Explain the biochemical principle behind the separation of
carbohydrates molecules by TLC. ii) Aldohexose sugars can form 16 different stereoisomers. How
many of those isomers can be distinguished and resolved by TLC and why? (no more than 100
words; use key words) (1 mark)
(i)Two separate phases exist at TLC. Stationary phase and mobile phase. Carbohydrates interact
differently with each other's stationary phase, which is why they are eluted at different times and run
at different speeds on TLC, making it easier to recognize them.
A rapid method is used to isolate carbohydrates on silica gel G combined with a small amount of
sodium bisulfite by means of TLC. The carbohydrates gave the reagents characteristic colours and
thus encouraged recognition.
(ii) We will get 8 aldohexoses separately, as D and L cannot be separated from TLC
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Ghebregzabher, M. et al. (1976) ‘Thin-layer chromatography of carbohydrates’, Journal of
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Chromatography A. Elsevier, 127(2), pp. 133–162.
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2. Sugars can constitute essential components of lipid molecules. (1 mark)

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i) What is the name of the shown molecule?

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ii) Where in biology can this molecule be found?
(i) Glucosylceramide
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(ii)A sugar sphingolipid consisting of a sphingoid backbone, a fatty acid, and a glucose moiety. Especially, GlcCer is
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present in plants, fungi , and animals and is absent in bacteria and certain eukaryotes such as Saccharomyces
cerevisiae baker yeast.
(iii)It is a major constituent of skin lipids, where it is important in the stratum corneum for lamellar body formation
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and in order to preserve the skin's water permeability barrier. Furthermore, the epidermal glucosylceramides
(together with sphingomyelin) are the source of the unusual complex ceramides found in the stratum corneum,
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including those with terminal hydroxyl groups and fatty acids associated with estolides. Part of the glucosylceramide
in skin is covalently connected to proteins by terminal hydroxyl groups, possibly to strengthen the epidermal barrier.
iv) name is N-nervonylglucosylsphingosine where anomer is sphingosine and isomer is Beta isomer and the ring is
glucose ring. https://pubchem.ncbi.nlm.nih.gov/compound/C24_1-GlcCer
Lavie, Y. et al. (1996) ‘Accumulation of glucosylceramides in multidrug-resistant cancer cells’, Journal of Biological
Chemistry. ASBMB, 271(32), pp. 19530–19536.
iii) What is the function of the molecule?

iv) Provide the precise chemical description (name, anomer, isomer and ring form) of the
sugar component. (1 mark)
Sugars are integral components of nucleotides and nucleic acids.
a) Provide the precise chemical description (anomer, isomer and ring form) of circular stereoisomers
that are possibly formed by 2’ deoxyribose in cells (1 mark)
2’ deoxyribose has a furanose ring in which there is no -OH group at 2 position of
carbon. Its isomer is aldopentose. It is a monosaccharide having 5 carbons and
aldehyde as a functional group.
Aldehyde carbon is an anomeric carbon of 2-deoxyribose.
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b) Using the language of the unit, provide a precise description of all chemical bonds that involve
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sugar moieties within a linear polynucleotide strand that carries a phosphate group at the 5’ end
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and a free 3’ OH group (1 mark)
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Cyclic form of aldopentose which is isomer is furanose in which 1’ carbon (anomeric
carbon/aldehyde carbon) is binding with base(purine/pyrimindine)
5’ carbon binds with phosphate group (carbon-oxygen bond)

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3’OH group is free and doesn’t bind in furanose form.

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c) Using the language of the unit, give an explanation whether the sugar components of RNA are
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reducing sugars? In your answer include the reducing sugar chemistry (no more than 100 words;
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use key words). (1 mark)
All mono carbohydrates are reducing sugar whether they are aldo or keto but there
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lies an exception like di carbohydrates have a non-reducing sugar like sucrose.

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Reducing sugar means that -OH group is free on anomeric carbon in cyclic form so
sugar components of RNA are non-reducing sugar because its furanose ring doesn’t
have free -OH group
https://pubchem.ncbi.nlm.nih.gov/compound/635

4. Could the purity of cellulose be estimated using the same method, acid hydrolysis, as used to
estimate the purity of glycogen during practical 5? (no more than 100 words; use key words)
(1 mark)
No, it could be as glycogen and starch both belong to polysaccharides. Glycogen is a

storage form of glucose in animals. its monomer is alpha-D-glucose with C1-C4
glycosidic bond. branching is introduced through C1-C6 linkages in the molecules.
As the linkage is alpha-glycosidic link which is easily hydrolysed by dilute HCl giving
individual monomer units hence leading to purity of Glycogen by acidic hydrolysis.
In case of starch, it is storage form of glucose in plants having Beta-D- glucose as its
monomer units. The glycosidic linkage in cellulose in beta and connected through C1-
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C4. therefore linkages are much stronger and remain in intact in presence of dilute HCl,
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therefore the purity of cellulose cannot be estimated using the acid hydrolysis method.
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Grossman, R. F. and Nwabunma, D. (2013) Biopolymer nanocomposites: processing,
properties, and applications. John Wiley & Sons.
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5.) A 1 mL sample of glycogen was calculated to contain 32 µmol (micromole) glucose. To 1 mL of this
sample was added 2 mL of 2 M HCl. It was then hydrolysed by boiling the solution for 15 minutes.
After boiling the hydrolysate was cooled and made up with H 2O to a final volume of exactly 10 mL.
The glucose was measured in this solution and found to have a concentration of 470 µg/mL
(microgram/milliliter).
Calculate the purity of the glycogen used in the sample as

% Purity = (moles of measured glucose/ moles of calculated glucose in glycogen) *100
Show your working out such that the marker can easily understand it. Include all units. Assume the
molecular mass of glucose is 180 g/mol and that the molecular mass of glucose within glycogen is
162 g/mol. At each step round off your calculations to 1 decimal point. (1 mark)
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As Final Volume of hydrolysate solution = 10 mL
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Measured Concentration of Glucose = 470 µg/mL (microgram/milliliter) = 470 × 10-6 g / mL
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Molar Mass of glucose within glycogen = 162 g/mol
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So,Moles of measured glucose = Measured Concentration of Glucose × Final Volume of
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hydrolysate solution / Molar Mass of glucose within glycogen
= 470 × 10-6 g / mL × 10 mL / 162 g / mol
= 29.0 × 10-6 mol
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Also, Moles of calculated glucose in glycogen

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= 32.0 µmol = 32.0 × 10-6 mol

Percent Purity , % Purity = ( Moles of measured glucose / Moles of calculated glucose in
glycogen ) × 100
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= ( 29.0 × 10-6 mol / 32.0 × 10-6 mol ) × 100 = 90.6 %

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6.) Production of cheap beer sometimes utilises sugarcane rather than malted barley. i) Research the
major sugar components of these starting materials and ii) Devise a simple experiment with suitable
controls that allows you to conclude whether a beer was and was not produced from sugar cane or
malted barley. Explain the carbohydrate chemistry behind the test to justify your answer. (1.5 mark)
In addition to general laboratory equipment the ONLY reagents available to you are:
- a beer sample
- the enzymes invertase and maltase
- the Somogyi-Nelson reagents for the determination of reducing sugars.
(i) Water: 69-75 % ; Non-reducing Sugar (8-16 %) ; Reducing Sugar (0.5 -2.0 % )
Sugar composition malted-barley: 76.13 % maltose, 15.81 % glucose, 6.3 %
sucrose, 2.04 % fructose and the remainder is lactose.
(ii) Maltose ===(Maltase)==> Glucose
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Sucrose==(invertase)==> Glucose + Fructose
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 Without fermentation, maltase is added to malt and Nelson-Somogyi process is used
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to assess sugar reduction.
 Without fermentation, add Invertase to sugar cane juice and use the Nelson-
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Somogyi method to determine reduction of sugars.
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 Non-beer solution by Nelson-Somogyi method will give postive test and high
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quantity to reduce sugars.
 After Beer formation (alcohol): these tests will be negative and the amount of
Nelson-Somogyi process reduction of sugars will be negligible.
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Hatanaka, C. and Kobara, Y. (1980) ‘Determination of glucose by a modification of

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Somogyi-Nelson method’, Agricultural and Biological Chemistry. Japan Society for

Bioscience, Biotechnology, and Agrochemistry, 44(12), pp. 2943–2949.
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SLE212 Biochemistry T1 2020 Assessment Worksheet D

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

2. Sugars can constitute essential components of lipid molecules. (1 mark)

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

i) What is the name of the shown molecule?

ii) Where in biology can this molecule be found?

iii) What is the function of the molecule?

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

Sugars are integral components of nucleotides and nucleic acids.

Aldehyde carbon is an anomeric carbon of 2-deoxyribose.

5’ carbon binds with phosphate group (carbon-oxygen bond)

3’OH group is free and doesn’t bind in furanose form.

use key words). (1 mark)

lies an exception like di carbohydrates have a non-reducing sugar like sucrose.

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

No, it could be as glycogen and starch both belong to polysaccharides. Glycogen is a

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

Calculate the purity of the glycogen used in the sample as

Also, Moles of calculated glucose in glycogen

= 32.0 µmol = 32.0 × 10-6 mol

= ( 29.0 × 10-6 mol / 32.0 × 10-6 mol ) × 100 = 90.6 %

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

Hatanaka, C. and Kobara, Y. (1980) ‘Determination of glucose by a modification of

Somogyi-Nelson method’, Agricultural and Biological Chemistry. Japan Society for

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

DO NOT MAKE CHANGES TO THE WORKSHEET TEMPLATE,

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