SUGGESTED ANSWER FOR TUTORIAL 6
QUESTION 1
a) For each of the following analysis, discuss the reason for the underlined parameter/
approach used.
i) In the solid phase extraction (SPE) process for the extraction of chlorphyrifos using
C18 sorbent, column conditioning was performed before sample application.
(4 marks)
C18 is a non polar sorbent which can retain the non polar chlorphyrifos.
Column conditioning to activate the solid sorbent i.e promote optimum
environment for retention (solvent composition, ionic strength, pH). Solvent
used should not interact with the sorbent.
ii) Fragrance analytes from plant sample were extracted using headspace-solid phase
microextraction (HS-SPME) followed by desorption process.
(4 marks)
Fragrance consists of volatile analytes. High temperature was utilized to
vaporize the volatile analytes in the sample and provide a headspace condition
above the sample for the adsorption.
Desorption can be achieved by placing the SPME fibre directly into GC injector
at high temperature (thermal desorption).
iii) Pressurized liquid extraction (PLE) is used to extract solutes from solid and semi
solid samples by utilizing high temperature and pressure.
(2 marks)
High temperature can disrupt the strong solute-matrix interactions and
decreases the viscosity of organic solvents, allowing improved penetration of
the matrix and thus improved extraction.
b) For each of the following, give a reason to the mentioned problem and suggest an
approach to overcome the problem without changing the extraction method used.
i) Alkanes can be extracted from urban air particulates using supercritical fluid
extraction (SFE) with CO2 at 75 bar (45C), whereas the polyaromatic
hydrocarbons (PAHs) remain unextracted.
(3 marks)
CO2 at 75 bar (45C) has low density therefore could not dissolve higher
molecular weight molecules. Therefore, increase the pressure. As the
pressure in the system is increased, density of CO2 increases and able to
dissolve and extract the PAHs.
Or
CO2 is non polar therefore not able to extract PAHs which are slightly more
polar than alkanes. Add a small amount of polar modifier into CO2 to
increase its polarity and enhance the solvating power. Thus PAHs can be
extracted.
ii) Non-polar contaminants from soil sample cannot be extracted using hexane with
microwave extraction method (MAE).
(3 marks)
Suitable solvents for MAE are polar solvents with high dipole moment,
such as water, methanol, or acetone because they are able to absorb
microwave radiation and pass it on to the sample molecules in the form of
heat. Therefore in order to extract the non polar contaminants using MAE,
hexane can be mixed with acetone.
(Q6a OCT 2010)
QUESTION 2
a) One of the extraction approach of solid phase microextraction (SPME) is by placing the
SPME fibre in the headspace above the aqueous sample.
i) Give another approach of SPME for the extraction of less volatile analytes.
(2 marks)
Direct immersion.
ii) Give two benefits of using SPME compared to solid phase extraction (SPE)?
(4 marks)
Analytes is adsorbed onto the surface of the fibre, it is solventless, faster
extraction time, clean extract and do not need any clean-up or
concentration and analysis time very fast.
In SPE, by using solid adsorbent to retain the analytes, long analysis time,
still need organic solvent but it is reduced the organic comsumption,
extract clean but still need to be concentrated, analysis time not as fast as
SPME.
b) The amount of solvent required in pressurized liquid extraction (PLE) method is smaller
than that required in traditional extraction method such as soxhlet method. Explain.
(4 marks)
The high temperature and pressure increased the capability of solvent to
penetrate the sample matrix. The solubility and diffusity of the analytes are
increased by the increased temperature, thus making the extraction faster and
less solvent is required.
c) Non-polar contaminants from soil sample cannot be extracted using hexane with
microwave extraction method (MAE). Give a reason to this problem and suggest an
approach to overcome the problem without changing the MAE method.
(4 marks)
(same question in Q6b(ii) OCT 2010)
Suitable solvents for MAE are polar solvents with high dipole moment, such as
water, methanol, or acetone because they are able to absorb microwave radiation
and pass it on to the sample molecules in the form of heat. Therefore in order to
extract the non polar contaminants using MAE, hexane can be mixed with acetone.
(Q6 JAN 2012)
QUESTION 3
a) You are given a water sample contaminated with several relatively non polar pesticides.
Outline the steps required for an efficient a solid phase extraction technique. Please
suggest specific sorbent and solvent in each step.
(4 marks)
1. Column (non polar such as C18) must be conditioned using methanol or/and
water.
2. Sample loading
3. Inteference elution using polar solvent such as water
4. Elution of analytes using non polar solvent such as hexane.
b) In the analysis of herbicides in soil sample, a pressurized liquid extraction (PLE)
technique is proposed prior to LC analysis. PLE technique is able to achieve comparable
efficiency as Soxhlet technique but at a shorter analysis time. Explain.
(5 marks)
High temperature:
- can disrupt the strong solute-matrix interactions
- decreases the viscosity of organic solvents allowing improved penetration of the
matrix, and hence improved extraction.
High pressure:
- Solvent remains liquefied above their boiling point and allows the solvent to
penetrate the sample matrix.
(Q5d,e JUNE 2012)
QUESTION 4
You are required to analyze colorant and fragrance compounds from a selected plant sample.
Several extraction methods such as Soxhiet, liquid-liquid extraction (LLE), solid phase
extraction (SPE), solid phase microextraction (SPME), microwave assisted extraction (MAE)
and pressurized liquid extraction (PLE) can be utilized prior to chromatographic analysis.
However, only n-hexane is available in the laboratory.
a) Suggest the most efficient and rapid method suitable for the extraction of non volatile
and low polarity colorant compounds. Justify your answer by discussing why the other
methods are not selected.
(6 marks)
LLE (1) and SPE (1) are not suitable for solid sample.
SPME is not suitable for non volatile compounds (1).
Soxhlet is not selected because the extraction time is too long, up to several
hours and it consumes a very large amount of organic solvent (1).
MAE is not suitable because non polar solvent is not able to absorb the
microwave radiation. (1)
(Note: Maximum 4 marks from LLE to MAE).
Thus, PLE is the most suitable (1). By using PLE, n-hexane can be utilized at
higher temperature to increase its polarity. (1).
b) These colorants are used in textile dyeing and the wastes are disposed into natural
water. By using SPE method with pre-conditioned C18 sorbent, you can perform an
extraction method to determine low level of colorants in the effluent. Discuss the steps
involve in this process.
(9 marks)
1. Sample loading. - (1)
The non-polar colorant compounds will be retained by the C18 sorbent and
some of the interferences will pass through. -(2)
2. Eluting interferences using deionized water. -(1)
Deionized water will elute the retained polar interferences while the non polar
analytes remain on the sorbent. -(2)
3. Eluting analytes using non polar solvent. (1)
Non polar analytes will be removed by non polar solvent but leaving behind
any interferences retained onto the sorbent. (2)
c) Suggest a suitable method for the analysis of fragrance compounds. Justify your answer.
(4 marks)
SPME (1)
Fragrance compounds are volatile -(1)
SPME is clean/solventless method that suitable for volatile anaytes and can be
used with solid samples by using headspace approach. -(1)
Directly desorbed into GC column. (1)
(Q5 JAN 2013)
QUESTION 5
a) If you were to analyse traces of diesel from a piece of burned t-shirt, which sample
preparation method is more suitable, solid phase microextraction (SPME) or pressurized
liquid extraction (PLE)? Why and why not the other one?
SPME
SPME: volatile, use headspace, cleaner extract, solventless(Any 2)
PLE: not suitable for very volatile compounds and dirty sample, use solvent.(Any 2)
b) If you were to analyse chlorphyrifos in a wastewater sample, which sample preparation
metohod is more suitable, headspace SPME or solid phase extraction (SPE)? Why and
why not the other one?
SPE
SPE: non volatile in aqueous sample, achieve clean up and concentration (Any 2)
SPME: chlorpyrifos is not very volatile, not suitable for headspace.(Any 2)
c) If you were to analyse a non polar compound in soil, which sample preparation method
is more suitable, supercritical fluid extraction (SFE) or Soxhlet extraction? Why and why
not the other one?
SFE
SFE: Clean, environmental friendly, fast. (Any 2)
Soxhlet: High solvent consumption, dirty extract, long analysis time. (Any 2)
(Q6 JUN 2013)
QUESTION 6
In a sudden death case, a urine sample was taken for forensic analysis. In screening for drug
overdose, the recommended sample preparation methods include solid phase extraction (SPE)
and solid phase microextraction (SPME).
a) Explain why pressurized liquid extraction (PLE) is not suitable for the above analysis?
(2 marks)
The sample is a liquid, PLE is for solid samples.
b) In a SPE technique, discuss the importance of the type of sorbent.
(3 marks)
Must be able to retain the analytes of interest, therefore same polarity as the
analytes.
c) Discuss the criteria of solvent chosen for each of the following step:
i) Elution of interference,
ii) Elution of analyte.
(5 marks)
i) Solvent must be able to remove interference but not analyte from the
sorbent.
ii) Solvent must be able to selectively extract the analytes from the sorbent.
Unwanted compounds should be retained on the sorbent.
d) Explain why headspace SPME is preferable compared to direct immersion SPME for the
above analysis.
(2 marks)
Using the HS-SPME, the headspace sample is cleaner, proteins and other non-
volatile compounds remain in the liquid matrix.
e) Discuss one limitation in the application of headspace SPME.
(2 marks)
Only volatile drug will be extracted, non volatile will remain in the liquid phase.
(Q5 DEC 2013)
QUESTION 7
a) On the determination of pesticides in soil samples, pressurized liquid extraction (PLE) is
a preferred method compared to soxhlet extraction due to better efficiency and faster
analysis.
i) Compare the two methods in terms of extraction time and solvent consumption
(include an approximate extraction time and volume of solvent)
(6 marks)
PLE: uses less solvent (1), 30 -50 mL (1)
Soxhlet: 200 -250 mL (1)
PLE: extraction time is shorter (1), 15 min (1)
Soxhlet: 12-18 hours (1)
ii) Discuss the effect of temperatue on the efficiency of PLE.
(3 marks)
High temperature (1) can disrupt the strong solute-matrix interactions and
decrease the viscosity of organic solvents, allowing improved penetration
of the matrix and thus improved extraction (2)
b) Headspace solid phase microextraction (HS-SPME) is the suitable method for the
analysis of flavor compounds in coffee.
i) Discuss two (2) advantages of HS-SPME compared to solid phase extraction
(SPE) for the mentioned analysis.
(6 marks)
(Any TWO answers)
Volatile sample (1). If SPE is used, volatile analytes might be lost (2).
Solventless (1). SPE consumes a small volume of organic solvent (2).
Concentrated extract (1). SPME do not need preconcentration (1). SPE may
require preconcentration after extraction (1).
Fast analysis (1). SPME is a single step extraction (extraction,
preconcentration, desorption) (1). SPE involve four steps (1)/ faster
extraction in SPME due to the fast equilibration between extraction phase
and sample matrix (2).
ii) List four (4) experimental parameters that can affect the efficiency of HS-SPME.
(4 marks)
Extraction (adsorption) time, thickness of fiber coating, extraction
temperature and types of fiber.
c) Extraction of active compounds from Tongkat Ali root sample using microwave assisted
extraction (MAE) method with pentane as an extraction solvent was unsuccessful.
Explain.
MAE is suitable for polar solvent (1). Change the solvent (1) to methanol or water
or mixed pentane with acetone/diethyl ether (1) since polar solvent can absorb
microwave radiation and penetrate to sample matrix (2).
(Q5 June 2014)
