Session 10

Tutorial-III
1. The Fischer projection of glucose is given here. 1

2
a. Is this the L-glucose or D-glucose? 3
4
As the last chiral carbon (C5) has –OH group on 5
the right, it is a D-sugar. It is, therefore, D-glucose
6
b. Draw the structure of β-anomer of this sugar in the Haworth form (Haworth formula).

2 2
c. What is the stereochemistry
in terms of R/S nomenclature? 4 1 1 3
2
4 1 3 4
3
2. The structure of fructose in Haworth formula is given here.

a. Identify the anomeric carbon.

b. Identify the anomer (α/β).

The –OH on the anomeric carbon is
cis to the –CH2OH on C5. The sugar,
therefore, by definition, is β.

c. Convert it into the Fischer projection.

HOCH2 H OH H HOCH2 OH
O O
H CH2OH
H OH H OH
OH CH2OH H OH H CH2OH
O
OH H HOCH2 OH OH H
 CH2OH
HOCH2 O
OH
HOCH2 H OH O
H OH CH2OH
OH CH2OH
H OH H
H OH H O
OH H
OH
H

HOCH2 H

OH
CH2OH

O d. Number the carbon atoms and identify the

the stereochemistry of the last chiral carbon
OH H in terms of R/S nomenclature.
CH2OH
H
OH
H OH O
CH2OH 2 2
OH H
H 2 4 1 1 3
OH
4H OH 1 3 4

3 CH2OH
3. The structure of a sugar (disaccharide) is shown here.

a. Identify (name) the glycosidic linkage.

Identify the anomeric carbons.

Identify the anomers: α, α

The linkage is α, α(1→1)

b. Is the sugar reducing? Why/Why not?

No.

As there is no hemiacetal/hemiketal, the sugar is non-reducing.

4. Examine the Haworth projections shown on right to answer the
questions below.

a. Circle each anomeric carbon.

b. Which of the two is the α anomer and which is the β anomer?
c. Are these structures considered enantiomers or diastereomers?
Diastereomers

d. Are these monosaccharides reducing sugars? Explain.

Yes. They have hemiacetal carbons

e. Is it possible to convert α anomer to β anomer or vice versa? Explain.

Yes. Through mutarotation.

5. Look at the peptide shown below and answer the questions

𝜀𝑊,280 𝑛𝑚 = 5690 𝑀−1 𝑐𝑚−1 𝜀𝐹,280 𝑛𝑚 = 143 𝑀−1 𝑐𝑚−1

𝜀𝑌,280 𝑛𝑚 = 1280 𝑀−1 𝑐𝑚−1 𝜀𝐶𝑦𝑠𝑡𝑖𝑛𝑒,280 𝑛𝑚 = 300 𝑀 −1 𝑐𝑚−1

a. Identify the N and C-termini of the peptide?

a. What will be the net charge on the peptide at neutral pH?
+1
 𝜀𝑊,280 𝑛𝑚 = 5690 𝑀−1 𝑐𝑚−1 𝜀𝐹,280 𝑛𝑚 = 143 𝑀−1 𝑐𝑚−1
𝜀𝑌,280 𝑛𝑚 = 1280 𝑀−1 𝑐𝑚−1 𝜀𝐶𝑦𝑠𝑡𝑖𝑛𝑒,280 𝑛𝑚 = 300 𝑀 −1 𝑐𝑚−1

c. Determine the concentration of the peptide if the peptide gives an

absorbance of 0.2 at 280 nm in a 1-cm path length cell.

𝐴280 𝑛𝑚 = (𝑛𝑊 𝜀𝑊 +𝑛𝑌 𝜀𝑌 +𝑛𝐹 𝜀𝐹 +𝑛𝐶𝑦𝑠𝑡𝑖𝑛𝑒 𝜀𝑐𝑦𝑠𝑡𝑖𝑛𝑒 )𝑐𝑙

0.2 = 5690 + 2 × 1280 + 143 + 300 × 𝑐 × 1

0.2
𝑐= 𝑀 = 2.3 × 10−5 𝑀 = 23 𝜇𝑀
8693
6. A sample of RNA is hydrolyzed and separated into three fractions by
column chromatography. Two of the three fractions are pure
nucleotides, but the third contains both adenylic and guanylic acid. At
pH 7.0, the absorbance of the mixture is 0.305 at 280 nm and 0.655 at
250 nm in 1-cm cells. The molar extinction coefficients for each pure
component at pH 7.0 are:

𝐴280 = (𝜀𝐴,280 × 𝑐𝐴 ) + (𝜀𝐺,280 × 𝑐𝐺 ) × 𝑙 𝐴250 = (𝜀𝐴,250 × 𝑐𝐴 ) + (𝜀𝐺,250 × 𝑐𝐺 ) × 𝑙

0.305 = (2300𝑐𝐴 + 9300𝑐𝐺 ) 0.655 = (12300𝑐𝐴 + 15700𝑐𝐺 )

Solving two linear equation gives the concentration.

7. Coloured pH indicators are dyes that have different spectra for
different ionization states. Assume the ionization of a pH indicator with
pK = 6.00 can be written as:

𝐻𝐼𝑛 ⇌ 𝐼𝑛− + 𝐻 +

The measured absorbance in a 1-cm cell at an indicator concentration of

2 × 10-5 M is given in the table below.

For the same concentration of indicator, an absorbance of 0.100 is measured at λ =

400 nm.

a. What is the pH of the solution?

b. What is the absorbance at λ = 440 nm?
𝐻𝐼𝑛 ⇌ 𝐼𝑛− + 𝐻 + pK = 6.00
𝐼𝑛−
𝑝𝐻 = 𝑝𝐾 + 𝑙𝑜𝑔
𝐻𝐼𝑛

𝑇ℎ𝑒 𝑡𝑎𝑏𝑙𝑒 𝑠𝑢𝑔𝑔𝑒𝑠𝑡𝑠 𝑡ℎ𝑎𝑡 𝑎𝑏𝑠𝑜𝑟𝑝𝑡𝑖𝑜𝑛 𝑎𝑡 400 𝑛𝑚 𝑖𝑠 𝑒𝑥𝑐𝑙𝑢𝑠𝑖𝑣𝑒𝑙𝑦 𝑑𝑢𝑒 𝑡𝑜 𝐻𝑖𝑛

while that at 460 and 480 nm is due to In–. Therefore, we can write:

𝐴400 = 𝜀𝐻𝐼𝑛 𝑐𝐻𝐼𝑛 𝐴480 = 𝜀𝐼𝑛− 𝑐𝐼𝑛−

𝑁𝑜𝑤, 𝑡ℎ𝑒 𝑡𝑜𝑡𝑎𝑙 𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑡ℎ𝑒 𝑖𝑛𝑑𝑖𝑐𝑎𝑡𝑜𝑟, 𝑐𝑇 = 𝐻𝐼𝑛 + 𝐼𝑛−

At pH 3, 𝑐𝑇 = 𝑐𝐻𝐼𝑛 At pH 9, 𝑐𝑇 = 𝑐𝐼𝑛−

𝐴400, 𝑝𝐻 3 = 𝜀𝐻𝐼𝑛 𝑐𝑇 𝐴480, 𝑝𝐻 9 = 𝜀𝐼𝑛− 𝑐𝑇

𝐴400 𝜀𝐻𝐼𝑛 𝑐𝐻𝐼𝑛 [𝐻𝐼𝑛] 𝐴480 𝜀𝐼𝑛− 𝑐𝐼𝑛− [𝐼𝑛− ]

= = = =
𝐴400, 𝑝𝐻 3 𝜀𝐻𝐼𝑛 𝑐𝑇 𝑐𝑇 𝐴480, 𝑝𝐻 9 𝜀𝐼𝑛− 𝑐𝑇 𝑐𝑇

[𝐼𝑛− ] 𝐴480 𝐴400, 𝑝𝐻 3

= ×
[𝐼𝑛𝐻] 𝐴480, 𝑝𝐻 9 𝐴400
[𝐼𝑛− ]
Measurements of absorbance therefore allows to determine
[𝐼𝑛𝐻]
For the same concentration of indicator, an absorbance of 0.100 is
measured at λ = 400 nm.
a. What is the pH of the solution?

As 𝐴400 = 0.1, i.e. the 50% of the value observed at pH 3.

[𝐼𝑛− ]
=1
[𝐼𝑛𝐻]
𝐼𝑛−
𝑝𝐻 = 𝑝𝐾 + 𝑙𝑜𝑔
𝐻𝐼𝑛
𝑝𝐻 = 𝑝𝐾 + 𝑙𝑜𝑔1

𝑝𝐻 = 𝑝𝐾 =6

b. What is the absorbance at λ = 440 nm?

As the absorbance of 𝐻𝐼𝑛 and 𝐼𝑛− is same at 440 nm and the

concentrations of 𝐻𝐼𝑛 and 𝐼𝑛− are same in the given solution
𝐴440, 𝑝𝐻 3 𝐴440, 𝑝𝐻 9 0.15 0.15
𝐴= + = + = 0.15
2 2 2 2