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Analytical 3 Assignment 1

This document contains a student's assignment responses for an analytical chemistry course. The assignment addresses topics including: [1] Isotope dilution analysis and its application to determining the weight of tryptophan in a protein hydrolysate sample. [2] Ways to manipulate retention factor and selectivity factor in partition chromatography, including changing mobile phase composition, column temperature, stationary phase properties. [3] Differences between electron impact, field ionization, and chemical ionization mass spectrometry ion sources and why double-focusing mass spectrometers give higher resolution than single-focusing models. [4] Challenges of coupling liquid chromatography to mass spectrometry compared to gas chromatography-

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Mike Vhurinoshara
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71 views

Analytical 3 Assignment 1

This document contains a student's assignment responses for an analytical chemistry course. The assignment addresses topics including: [1] Isotope dilution analysis and its application to determining the weight of tryptophan in a protein hydrolysate sample. [2] Ways to manipulate retention factor and selectivity factor in partition chromatography, including changing mobile phase composition, column temperature, stationary phase properties. [3] Differences between electron impact, field ionization, and chemical ionization mass spectrometry ion sources and why double-focusing mass spectrometers give higher resolution than single-focusing models. [4] Challenges of coupling liquid chromatography to mass spectrometry compared to gas chromatography-

Uploaded by

Mike Vhurinoshara
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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BINDURA UNIVERSITY OF SCIENCE EDUCATION

FACAULTY OF SCIENCE

CHEMISTRY DEPARTMENT

STUDENT NAME : MIKE VHURINOSHARA

REG NUMBER : B1748924

LEVEL : 4.1

COURSE CODE : CH 304

COURSE DESCRIPTION : ANALYTICAL CHEMISTRY 3

LECTURER : MR MUSEKIWA

DUE DATE : 20/11/2020

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1. (a) Discuss the principles of isotope dilution analysis (IDA) in radiochemical
methods.

This is a form of internal standardization in which a radioactive form of analyte serves as the
internal standard. In this method, a tracer (internal standard) of known activity is
prepared and a measured mass of tracer (Wt) is added to a sample containing an unknown
mass wx of non-radioactive analyte. The mixture is homogenized (same phase mixture)
and the sample is then processed to isolate weight in grams of purified analyte,
containing both radioactive and non-radioactive material. The activity of isolated sample
is measured (AA). The activity of isolated sample AA is calculated as follows:

AT
w x= w −wT
AA A

(b) To sample of a protein hydrolysate, a chemist added 1.00 mg of tryptophan,


which was labeled with 14C and exhibited a counting rate of 584 counts per minute
(cpm) above background. After this labeled compound was thoroughly mixed with
the sample, the mixture was passed through an ion – exchange column. The fraction
of the effluent containing only tryptophan was collected, and from it an 18.0 mg
sample of pure tryptophan was isolated. The isolated sample had a count of 204 cpm
in the same container. What was the weight of tryptophan in the original sample?

AT
w x= w −wT
AA A

584 cpm
w x= × 18.0 mg−1.0 mg
204 cpm

¿ 48.67 mg

2. (a) Describe ways to manipulate both the retention factor (k’) and the selectivity
factor (gα) in partition chromatography.

Selectivity Retention factor


Adjusting the column temperature for gas Changing column length
chromatography
Changing the composition of the mobile Temperature programming in gas
phase(gradient elution) for liquid chromatography
chromatography
Using different chemical reagents, ion Changing nature of the stationary phase
pairing, complexing or surfactants
Changing the composition of the The pressure used which affect the flow

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stationary phase rate of the solvent

(b)(i) For a normal - phase separation, predict the order of elution of n – hexane,
n – hexanol, benzene.

n hexane>benzene>n hexanol

(ii) For a reversed – phase separation; predict the order of elution of ethyl acetate,
diethyl ether, and nitrobenzene.

Diethyl ether >ethyl acetate>nitro butane

(c). On a silica gel column, a compound was found to have a retention time of 28
min when the mobile phase was toluene. Which solvent, carbon tetrachloride or
chloroform, would be more likely to shorten the retention time? Explain.

Chloroform because it is more polar than carbon tetrachloride. The extent to which a
compound is retained will depend primarily upon its polarity, in the case of silica gel, and
primarily upon its lipophilicity in the case of a reversed-phase packing.

3. (a) How do the spectra for electron - impact, field ionization and chemical
ionization sources differ from one another?

The most fragmentation and thus the complex spectra are encountered with electron
impact ionization. Field ionization produces the simplest spectra. Chemical and electron
impact ionization results in higher sensitivities than does field ionization.

(b) Why do double – focusing mass spectrometers give narrower peaks and higher
resolutions than their single – focusing mass spectrometer counterparts?

The resolution of a single focusing mass spectrometer is limited by the initial kinetic
energy spread of the sample molecules. The spread is minimized in a double focusing
instrument by accelerating the sample through an electrostatic analyzer, which limits the
range kinetic energies of ions being introduced into the magnetic sector analyzer.
Significantly narrower peaks are a result

(6 marks)

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(c) Discuss why the coupling of liquid chromatography to a mass spectrometer
presents more challenges than the coupling of a gas chromatography to a mass
spectrometer

Flow rates of 0.5 to 1.5 mL/min in conventional wide-bore columns generate a large gas
flow too large for MS vacuum system.

Common HPLC solvents and non-volatile additives are incompatible with LC/MS
systems.

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