EXTRACTION OF

VOLATILE COMPOUNDS
 VOLATILE ORGANICS AND THEIR ANALYSIS

From an analytical point of view:

volatile organic compounds (VOCs) can be defined as organic compounds whose
vapor pressures are greater than or equal to 0.1 mmHg at 20C.
◦ An important feature of VOC analysis is that in most cases the
analytes are first transferred to a gas–vapor phase and then
analyzed by an instrument.
◦ Gas chromatography (GC) is the instrumental method of choice
for the separation and analysis of volatile compounds. GC is
mature, extremely reliable, and there is a wealth of literature
regarding analysis of volatile compounds by GC
◦ In general, the analysis of pure volatile compounds is not difficult
and can be accomplished via direct injection of the analyte into
a gas chromatograph
◦ The challenge is to extract the analytes from this matrix
reproducibly, and to accurately determine their mass or
concentration. There are several approaches to this, including
static headspace extraction (SHE), dynamic headspace
extraction (purge and trap), solid-phase micro- extraction (SPME)
◦ The choice of technique depends on the type of sample matrix,
information required (quantitative or qualitative), sensitivity
required, need for automation, and budget.
STATIC HEADSPACE EXTRACTION
STATIC HEADSPACE EXTRACTION
◦ Static headspace extraction is also known as equilibrium headspace extraction or
simply as headspace.
◦ It is one of the most common techniques for the quantitative and qualitative
analysis of volatile organic compounds from a variety of matrices.
◦ This technique has been available for over 30 years , so the instrumentation is both
mature and reliable. With the current availability of computer-controlled
instrumentation, automated analysis with accurate control of all instrument
parameters has become routine.
◦ The method of extraction is straight forward: A sample, either solid or liquid, is
placed in a headspace autosampler (HSAS) vial, typically 10 or 20 mL, and the
volatile analytes diffuse into the headspace of the vial . Once the concentration
of the analyte in the headspace of the vial reaches equilibrium with the
concentration in the sample matrix, a portion of the headspace is swept into a
gas chromatograph for analysis. This can be done by either manual injection or by
use of an autosampler.
 Sample Preparation for Static
Headspace Extraction
◦ For qualitative analysis, the sample can be placed directly into the headspace vial and
analyzed with no additional preparation.
◦ However, for quantitation, it may be necessary to understand and optimize the matrix effects to
attain good sensitivity and accuracy.
◦ For quantitative analysis of volatile compounds from solid particles, equilibrium between the
analyte concentration in the headspace and in the sample matrix must be reached in a sensible
period of time, typically a matter of minutes.
◦ For large solid samples it may be necessary to change the physical state of the sample matrix.
◦ Two common approaches are crushing or grinding the sample and dissolving the solid into a
liquid.
◦ The first approach increases the surface area available for the volatile analyte to partition into
the headspace. However, the analyte is still partitioning between a solid and the headspace.
◦ The second approach is preferred since liquid or solution sample matrices are generally easier to
work with than solids since the analyte partitioning process into the headspace usually reaches
equilibrium faster. Also, analyte diffusion in liquids eliminates unusual diffusion path problems,
which often occur with solids and can unpredictably affect equilibration time.
 Optimizing Static Headspace Extraction Efficiency
and Quantitation

◦ There are many factors involved in optimizing static headspace extraction for
extraction efficiency, sensitivity, quantitation, and reproducibility.
◦ These include vial and sample volume, temperature, pressure, and the form of the
matrix itself, as described above.
◦ The appropriate choice of physical conditions may be both analyte and matrix
dependent, and when there are multiple analytes, compromises may be necessary.
◦ The major factors that control headspace sensitivity are the analyte
partition coefficient (K) and phase ratio 𝛃.

◦ where A is the GC peak area for the analyte, CG the concentration

of the analyte in the headspace, C0 the initial concentration of the
analyte in the liquid sample, K the partition coefficient, and b the
phase volume ratio.
◦ The effect of the parameters K, controlled by the extraction
temperature.
◦ and 𝛃, controlled by the relative volume of the two phases,
◦ on static headspace extraction analysis sensitivity depends on the
solubility of the analyte in the sample matrix.
◦ Sensitivity is increased as β is minimized , that is lower value ( larger sample sizes) will
yield higher responses for volatile compounds .
◦ However, changes in 𝛃 will not always yield the increase in response needed to
improve sensitivity ;
◦ When 𝛃 is decreased by increasing sample size , compound with higher K value
partition less into the headspace compared to compounds with low K value and yield
correspondingly smaller changes in Cg .
◦ Samples that contain compounds with high K need to be optimized to provide the
lowest K before changes are made in 𝛃
◦ For analytes that have a high partition coefficient, temperature
will have a greater influence than the phase ratio. (This is because the
majority of the analyte stays in the liquid phase, and heating the vial drives the volatile
into the headspace).

◦ For volatile analytes with a low partition coefficient, The volumes

of sample and headspace have a greater influence on sensitivity
than does the temperature (the majority of the volatile analyte is already in
the headspace of the vial).
1. Ethanol
2. Methylethylketone
3. Toluene
4. Hexane
5. Tetrachloroethylene
◦ For a polar analyte in an aqueous matrix, the sample volume will have minimal effect on
the area response and a dramatic effect on less polar analytes.

Analysis of three samples of an aqueous

solution of :
cyclohexane (0.002 vol %) and
1,4-dioxane (0.1 vol %)
in a 22.3-mL vial:
(a) 1.0 mL of solution ( 𝛃= 21:3);
(b) 5.0 mL of solution (𝛃= 3:46);
(c) 5.0 mL of solution (𝛃= 3:46) to which 2 g
of NaCl was added.
Head- space conditions: equilibration at
60C, with shaker.

Peaks: 1, cyclohexane; 2, 1,4-dioxane.

Quantitative Techniques in Static
Headspace Extraction
◦ The most common approaches to quantitative HSGC calibration
are:
1. classical external standard,
2. internal standard,
3. standard addition
External Standard Calibration
External standard quantitation involves the preparation of
a classical calibration curve.
Standard samples are prepared at various concentrations
over the desired range and analyzed. A calibration curve
is then generated, with raw GC peak area plotted versus
standard concentration.
Peak areas of each analyte are then determined and
compared to the curve to generate analyte
concentration.
This method is best for analytes in liquid samples where the
analytes are soluble in the sample matrix and the matrix
has no effect on the analyte response.
If the analyte has a low solubility in the sample matrix,
preparation of standards via serial dilution can be difficult.
It is important to match the standard and sample matrix as
closely as possible and to demonstrate equivalence in the
response between the standards and samples.
Internal Standard Calibration
Internal standard calibration can be used to compensate for
variation in analyte recovery and absolute peak areas due to
matrix effects and GC injection variability.
Prior to the extraction, a known quantity of a known additional
analyte is added to each sample and standard. This
compound is called an internal standard. To prepare a
calibration curve the standards containing the internal
standard are chromatographed. The peak areas of the
analyte and internal standard are recorded. The ratio of areas
of analyte to internal standard is plotted versus the
concentrations of the known standards. For the analytes, this
ratio is calculated and the actual analyte concentration is
determined from the calibration graph.
Although internal standard calibration compensates for some
errors in external standard quantitation, there are several
difficulties in method development. First, choosing an
appropriate internal standard can often be difficult, as this
compound must be available in extremely pure form and it
must never appear in the samples of interest. Second, it cannot
interfere in either the extraction or the chromatography of the
analytes. Finally, it must be structurally similar to the analytes, so
that it undergoes similar extraction and chromatography,
otherwise, the compensation will be lost.
Standard Addition
In standard addition calibration, an additional
known quantity of the analyte is added directly to
the samples, following an initial analysis. By adding
one or more aliquots of standard, a calibration
curve can be prepared.
The concentration of analyte in the sample can
then be determined by extrapolating the
calibration curve. For this method, analyte response
must be linear throughout the range of
concentrations used in the calibration curve. A
practical approach to standard addition is to
divide up the sample into several equal portions,
then add increasing levels of standard. The final
concentration of the standard is the concentration
of the standard after it is added to the sample. The
original concentration is then determined by
extrapolation to the x-axis.
DYNAMIC HEADSPACE EXTRACTION OR PURGE AND
TRAP
◦ For the analysis of trace quantities of analytes, or where an exhaustive extraction
of the analytes is required, purge and trap, or dynamic headspace extraction, is
preferred over static headspace extraction.
◦ Like static head- space sampling, purge and trap relies on the volatility of the
analytes to achieve extraction from the matrix. However, the volatile analytes do
not equilibrate between the gas phase and matrix. Instead, they are removed
from the sample continuously by a flowing gas.
◦ This provides a concentration gradient, which aids in the exhaustive extraction of
the analytes.
◦ Purge and trap is used for both solid and liquid samples, which include
environmental (water and soil) , biological industrial, pharmaceutical, and
agricultural samples.
◦ This technique is used in many standard methods approved by the EPA .
 Instrumentation
The trap is usually a stainless steel tube 3 mm in inside
diameter (ID) and 25 mm long packed with multiple layers
of adsorbents The sorbents are arranged in layers in
increasing trapping capacity. During
purging/sorption, the purge gas reaches the weaker
sorbent first, which retains only less volatile species.
More volatile species break through this layer and
are trapped by the stronger adsorbents.
Desorption Adsorption
◦ Tenax is a porous polymer resin based on 2,6-diphenylene oxide.
It is hydrophobic and has a low affinity for water. However, highly
volatile compounds and polar compounds are poorly retained
on Tenax.
◦ Silica gel is a stronger sorbent than Tenax. It is hydrophilic and
therefore an excellent material for trapping polar compounds.
However, water is also retained.
◦ Charcoal is another sorbent that is stronger than Tenax. It is
hydrophobic and is used mainly to trap very volatile compounds
(such as dichloro/difluromethane, that can break through Tenax
and silica gel.
 SOLID-PHASE MICROEXTRACTION
◦ SPME is a solventless extraction method that employs a fused silica
fiber coated with a thin film of sorbent, to extract volatile analytes
from a sample matrix.
◦ The fiber is housed within a syringe needle that protects the fiber
and allows for easy penetration of sample and GC vial septa. Most
published SPME work has been performed with manual devices,
although automated systems are also available.
◦ There are two approaches to SPME sampling of volatile organics:
direct and headspace. In direct sampling the fiber is placed directly
into the sample matrix, and in headspace sampling the fiber is
placed in the headspace of the sample
SPME advantages
◦ SPME has several advantages in the analysis of volatile organics.
1. no additional instruments or hardware are required.
2. The cost of fibers is low compared to the cost of other methods for volatile analyte
extraction.
3. Fibers can be reused from several to thousands of times, depending on extraction
and desorption conditions.
4. SPME requires minimal training to get started, although there may be many variables
involved in a full-method development and validation.
5. SPME is also easily portable, and field sampling devices are readily available.
6. Finally, with a variety of fiber coating chemistries available, SPME can be applied to
a wide variety of volatile organic analytes.
 SPME Method Development for
Volatile Organics
◦ The key issues involved in developing an extraction procedure include:
1. extraction mode (direct or head- space),
2. choice of fiber coating,
3. agitation method,
4. length of extraction,
5. extraction temperature,
6. and matrix modification.
Extraction mode (direct or head- space),

◦ Choosing between direct immersion SPME and headspace SPME

is relatively straight forward. Direct immersion SPME is warranted
for liquid samples or solutions for which other solid-phase or liquid–
Liquid extraction methods would be considered.
◦ Headspace SPME would be considered for the same analytes as
static headspace extraction or purge and trap. Therefore,
headspace SPME should be considered for extracting volatile
compounds from solid or liquid samples, in which the normal
boiling point of the analyte(s) of interest is less than about 200C.
◦ For higher-boiling analytes, direct immersion SPME will probably
be necessary. Also, the nature of the sample matrix should be
considered. Headspace SPME is preferred for especially complex
or dirty samples, as these may foul the fiber coating in a direct
immersion analysis. However, SPME fibers have been shown to be
usable for about 50 direct immersions into urine . Some
laboratories have reported using a fiber for thousands of
extractions from drinking water.
Choosing an SPME Fiber Coating

◦ SPME fibers have different coatings for the same reason that GC
capillary columns have different coatings: There is no single
coating that will extract and separate all volatile organics from a
sample, therefore, different types of coatings with different
polarities are used on SPME fibers.
◦ Currently, three classes of fiber polarity coatings are
commercially available: nonpolar, semipolar, and polar coatings
Optimizing Extraction Conditions agitation
method, length of extraction, extraction temperature, and matrix
modification.

◦ Extraction time is optimized by extracting a standard using a range

of extraction times and plotting the analyte GC peak area versus
the extraction time. As extraction time is increased, a plateau in
peak area is reached. This represents the time required for the
system to reach equilibrium and is the optimized extraction time.
◦ The sample volume also has an effect on both the rate and
recovery in SPME extractions, as determined by extraction kinetics
and by analyte partition coefficients.
◦ As with any extraction, the agitation method will affect both the
extraction time and recovery and should be controlled as closely
as is practical.
◦ In direct-immersion SPME, agitation is usually accomplished using
magnetic stirring, so the stirring rate should be constant. Also, the
fiber should not be centered in the vial, as there is little to no
liquid velocity there; the fiber should always be off-centered so
that liquid is moving quickly around it. Agitation can also be
achieved by physical movement of the fiber or by movement of
the sample vial. Sonication is also used
◦ Extraction temperature can also be an important factor,
especially in headspace SPME analyses.
◦ The sample matrix may also be modified to enhance extraction
recovery.
◦ This is typically done by either dissolving a solid sample in a
suitable solvent, usually water or a strongly aqueous mixture, or
by modifying the pH or salt content of a solution.
◦ Modifying the pH to change the extraction behavior works the
same way in SPME as it does for classical liquid–liquid extraction.
At low pH, acidic compounds will be in the neutral form and will
be extracted preferentially into the fiber coating; at high pH,
basic compounds are extracted favorably. Neutral compounds
are not affected appreciably by solution pH.
Thank You