Code No: RR422301 Set No.

1
IV B.Tech II Semester Regular Examinations, Apr/May 2008
CHROMATOGRAPHIC SEPARATIONS
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
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1. Discuss the different processes that oppose the formation of a narrow analyte peak
to increase the plate height. [16]

2. From the distribution studies species M and N are known to have partition co-
efficients between water and hexane of 6.50 and 6.31. The two species are to be
separated by elution with hexane in a column packed with silica gel with water on
its surface. The ratio Vs/Vm for the packing is known to be 0.422.

(a) Calculate the capacity factor for each of the solutes.

(b) Calculate the selectivity factor.
(c) How many plates will be needed to provide a resolution of 1.5?
(d) If a flow rate of 7.10 cm/sec is employed, what time will be required to elute
the two species? [4×4=16]

3. (a) List out the commonly used ion exchangers and write matrices used for them.
(b) What are the factors that influence the ion exchange chromatography perfor-
mance? [8+8]

4. (a) Write a short notes on spot densitometer.

(b) Describe the application of TLC for separation of inorganic ions. [8+8]

5. What is the hydrophobic interaction chromatography? Explain with a neat dia-

gram. [16]

6. Write the principle, mode of operation of Gas chromatography. What kinds of

mixtures can be separated by gas chromatography? [16]

7. What is a racemic mixture? Discuss chromatographic method to separate a racemic

mixture. [16]

8. Write the principle and methods separation by capillary electrophoresis, and why
the pH affect the separation of amino acids in electrophoresis. [16]

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Code No: RR422301 Set No. 2
IV B.Tech II Semester Regular Examinations, Apr/May 2008
CHROMATOGRAPHIC SEPARATIONS
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. (a) What are the basic column chromatographic components? Explain.

(b) Explain the modes of chromatography in detail. [8+8]

2. Describe one method for the development of chromatogram and write a note on
column resolution. [16]

3. Briefly outline the advantages which may be gained by preparing specific ion ex-
change resins which contain functional groups known to react selectively with par-
ticular groups of cations. [16]

4. (a) Write a short notes on spot densitometer.

(b) Describe the application of TLC for separation of inorganic ions. [8+8]

5. What is the hydrophobic interaction chromatography? Explain with a neat dia-

gram. [16]

6. Write the essential principle of gas chromatography. Write the differences, advan-
tages and disadvantages of Gas Liquid Chromatography and Gas Solid Chromatog-
raphy. [16]

7. Write the principle of chiral chromatography. Write about its industrial applica-
tions. [16]

8. Write the principle of electrophoresis. Explain the components of capillary elec-

trophoresis instrumentation with a diagram. [16]

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Code No: RR422301 Set No. 3
IV B.Tech II Semester Regular Examinations, Apr/May 2008
CHROMATOGRAPHIC SEPARATIONS
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. Classify and explain the chromatographic techniques with suitable examples and
schematic diagrams. [16]

2. From the distribution studies species M and N are known to have partition co-
efficients between water and hexane of 6.50 and 6.31. The two species are to be
separated by elution with hexane in a column packed with silica gel with water on
its surface. The ratio Vs/Vm for the packing is known to be 0.422.

(a) Calculate the capacity factor for each of the solutes.

(b) Calculate the selectivity factor.
(c) How many plates will be needed to provide a resolution of 1.5?
(d) If a flow rate of 7.10 cm/sec is employed, what time will be required to elute
the two species? [4×4=16]

3. Explain the principle of ion-exchange chromatography with suitable examples. [16]

4. (a) What is the difference between thin layer chromatography and ion-exchange
chromatography? Explain.
(b) My friend Joe gave me 2g of a substance, which he claims is a lipid. He
also gave me a plain peace of paper, a beaker of water, 2 g of table sugar-and
nothing else. Describe in detail 4 simple tests .which would allow me to decide
whether the substance is indeed a lipid. In each of these tests, what would
the control be? [8+8]

5. (a) Write a short note a column efficiency and selectivity.

(b) Differentiate between HPLC and TLC. [8+8]

6. What is a chromatogram? Describe the physical differences between open tubular

and packed columns. [16]

7. Write short notes on:

(a) Enantiomers
(b) Chiral resolving agent
(c) Retention time
(d) Band broadening. [4×4=16]

8. (a) Write the principle of separation by capillary zone electrophoresis.

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Code No: RR422301 Set No. 3
(b) A inorganic cation has an electrophoretic mobilities of 4×10−4 cm−2 s−1 v−1 .This
same ion has diffusion coefficient of 10×10−6 cm2 .s−1 . If this ion is separated
by capillary zone electrophoresis with 50cm capillary,calculate the experted
plate count N at appliled voltage of 10kv. [8+8]

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Code No: RR422301 Set No. 4
IV B.Tech II Semester Regular Examinations, Apr/May 2008
CHROMATOGRAPHIC SEPARATIONS
(Bio-Technology)
Time: 3 hours Max Marks: 80
Answer any FIVE Questions
All Questions carry equal marks
⋆⋆⋆⋆⋆

1. (a) Define the effective distribution coefficient. What is the use of of distribution
coefficient in chromatography separations?
(b) What are the retention parameters? Discuss in brief. [8+8]
2. Discuss the single and variable wavelength detectors with suitable examples and
diagrams. [16]
3. What are the principles of separation of metals by ion exchange chromatography?
Explain. [16]
4. (a) How adsorbents are selected in TLC?
(b) Discuss the limitations of TLC. [8+8]
5. (a) Write briefly procedure for column packing.
(b) Write a short note on pumping systems used in HPLC. [8+8]
6. Write short notes on:
(a) Teperature programming in GC
(b) Band separation
(c) Electron capture detector
(d) Open tubular columns. [4×4=16]
7. Write short notes on:
(a) Enantiomers
(b) Chiral resolving agent
(c) Retention time
(d) Band broadening. [4×4=16]
8. (a) Write the principle of separation by capillary zone electrophoresis.
(b) A inorganic cation has an electrophoretic mobilities of 4×10−4 cm−2 s−1 v−1 .This
same ion has diffusion coefficient of 10×10−6 cm2 .s−1 . If this ion is separated
by capillary zone electrophoresis with 50cm capillary,calculate the experted
plate count N at appliled voltage of 10kv. [8+8]

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