1 / VOL. 2, NO. 4 THE CHEMICAL EDUCATOR 1997 SPRINGER-VERLAG NEW YORK, INC.

ISSN 1430-4171 http://journals.springer-ny.com/chedr 10.1007/s00897970137a

In the Classroom

Solid Phase Microextraction (SPME)

JANUSZ PAWLISZYN* Department of Chemistry University of Waterloo Waterloo, Ontario, N21 3G1 Canada bpawlisz@jeeves.uwaterloo.ca BARBARA PAWLISZYN AND MICHAEL PAWLISZYN JP Scientific Waterloo, Ontario, N2K 1W7 Canada

SPME methods are still in the initial stages of development, but the results are promising.

olid phase microextraction (SPME) was developed to address the need for a fast, solvent-free and field compatible sample preparation method. It has been applied to a range of applications including environmental, industrial hygiene, process monitoring, clinical, forensic, food and drug analysis. In SPME, coated fibers are used to isolate and concentrate analytes into a range of coating materials. After extraction, the fibers are transferred, with the help of the syringe-like handling device, to analytical instruments for separation and quantification of the target analytes.

In the past 20 years the number of instrumental techniques available to the chemist has grown exponentially. In order to help our readers keep up with this rapidly growing field, tutorial articles on chemical instrumentation will be a regular feature of The Chemical Educator. The articles are designed to serve as a brief introduction to emerging instrumental techniques with an outline of the underlying principles and major applications.

Martin Schimpf, Series Editor

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An analytical process typically consists of several discrete steps: sampling, sample preparation, separation, quantification and data analysis. For example, in the analysis of semivolatile components in water, the target analytes are first extracted into an organic solvent. The resulting solution is then introduced into an analytical instrument for separation, quantification, and possible identification. Each of these steps affect the precision, accuracy and speed of the analysis. Although multi-dimensional techniques such as gas chromatography/mass spectrometry (GC/MS) have improved separation and quantification, the preparation step is still time consuming and often uses a significant volume of organic. SPME was developed to simplify the preparation step. SPME is a microextraction technique, which means that the amount of extraction solvent is very small compared to the sample volume. As a result, exhaustive removal of analytes to the extracting phase does not occur, rather an equilibrium is reached between the sample matrix and the extracting phase. To make this approach practical, the extracting phase is permanently attached to rods made out of various materials. In most of the cases, the extracting phase is a polymeric organic phase that is cross-linked and permanently attached to the rod. In one configuration, the rods consists of an optical fiber made of fused silica, which is chemically inert. A polymer layer is used to protect the fiber against breakage. Two common polymers used are poly(dimethylsiloxane) and polyacrylate. Poly(dimethylsiloxane) behaves as a liquid, which results in rapid extraction compared to polyacrylate, which is a solid. The silica rods have a typical diameter of 100200 micrometers and a film thickness ranging from 10100 microns. When the coated fiber is placed into an aqueous matrix (Figure 1), the analyte is transferred from the matrix into the coating. The extraction is considered to be complete when the analyte has reached an equilibrium distribution between the matrix and fiber coating. The equilibrium condition can be described as: n= K fsV f Vs C0 K fsV f Vs (1)

when n is the amount extracted by the coating Kfs is the distribution coefficient between the fiber coating and the sample matrix, Vf is the volume of the fiber coating, Vs is the volume of the sample, and C0 is the initial concentration of analyte in the sample.

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FIGURE 1. MICROEXTRACTION WITH SPME.

Equation 1 indicates a proportional relationship between analyte concentration in the sample and the amount extracted by the coated fiber, which is the basis for analyte quantification. If the volume of the sample is very large compared to the volume of the coating, in other words, the amount of analyte extracted onto the fiber is an insignificant portion of that present in the sample, Equation 1 simplifies to n = K fsV f C0 (2)

In Equation 2, the amount of extracted analyte is independent of the volume of the sample. Therefore, there is no need to collect a defined amount of sample prior to analysis. Thus, the fiber can be exposed directly to the ambient air, water, production stream, etc., and the amount of extracted analyte will correspond directly to its concentration in the matrix. This greatly accelerates the analytical process, while errors associated with the loss of analyte through decomposition or absorption onto sampling-container walls is prevented.

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FIGURE 2. THE CUSTOM-MADE SPME DEVICE BASED ON HAMILTON 7000 SERIES SYRINGE.

Handling fragile fibers is very difficult, so to make the SPME approach practical, fibers have been incorporated in a micro syringe device, as shown in Figure 2. Movement of the plunger allows the fiber to be extruded from the needle for extraction or introduction into an analytical instrument. By moving the plunger up, the fiber is protected in the needle during both storage and penetration of injection-port septa. This approach has been commercialized by companies that sell chromatographic syringes, and has led to the modification of autosamplers by instrument manufacturers. SPME has several important advantages compared to traditional sample preparation techniques. The advantage of SPME can be illustrated by returning to the example discussed above. The SPME method for semivolatile analysis consists of inserting the fiber device into the aqueous sample matrix, pushing the plunger to expose the fiber, retracting the fiber into the needle when equilibrium has been reached, and finally introducing the fiber into the analytical instrument. During desorbtion of the analyte, the polymeric phase is cleaned and therefore ready for reuse. The absence of solvent in SPME is an important feature, as it is not only environmentally friendly but makes the

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separation faster, which increases throughput and allows for the use of simpler instruments. Another important feature of SPME is its small size, which is convenient for designing portable devices for field work. Since the amount of extracting phase is small, the equilibrium of the system is not disturbed and can therefore be studied. Very small objects can be studied with miniature fibers, such as a single flower or even a single cell. The sensitivity and limit of determination is comparable to techniques that rely on liquid extraction. Although only a small portion of analyte is extracted from the matrix, all extracted analytes are transferred to the analytical instrument. This is in contrast to liquid extractions, where the majority of analyte is transferred from a given sample to the organic phase but only a small portion (1/100 or 1/1000) of the extracted analyte is introduced to the analytical instrument. SPME can be used in two principle modes, that is direct-extraction and headspace configurations. Figure 3 illustrates the differences between these two modes. In the direct-extraction mode (Figure 3a), the coated fiber is inserted directly into the sample, and analytes are extracted directly from the sample matrix to the extraction phase. To facilitate rapid extraction, some level of agitation is required to enhance transport of the analyte from the bulk solution to the fiber. For gaseous samples, natural air convections and high diffusion coefficients are typically sufficient to facilitate rapid equilibration. For aqueous matrices, more efficient agitation techniques are required, such as forced flow, rapid fiber or vial movement, stirring or sonication. Agitation of liquid samples reduces the effect caused by depletion of the analyte zone next to the fiber. In the headspace mode (Figure 3b), the vapor above the bulk matrix is sampled. Thus, analytes must be relatively volatile in order to be transported from the bulk matrix to the fiber coating. Sampling of the headspace protects the fiber coating from damage by hostile matrices, such as those at very high or low pH, or those with large molecules, such as proteins and humic acids, which tend to foul the coating. Headspace sampling also allows for the analysis of solid matrices. The choice of sampling mode has a significant impact on extraction kinetics. When the headspace is sampled, the analytes are extracted from the matrix indirectly, as shown in Figure 3b. Therefore, since more volatile components are at a higher concentration in the headspace, they are extracted faster than less volatile components.

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FIGURE 3. MODES OF SPME OPERATION: (A) DIRECT EXTRACTION, (B) HEADSPACE SPME.

SPME methods are still in the initial stages of development, but the results are promising. Initial interest in SPME has been driven by the promise of a solvent-free environment, fast extraction, convenient automation and easy hyphenation with analytical instruments. So far, SPME methods have been developed primarily for the quantitative analysis of target analytes in relatively clean matrices. As the technology is developed, however, the capability for direct field measurements and the investigation of multiphase equilibrium processes should follow. Thus far, application work has been devoted primarily to environmental analysis, with a focus on aqueous

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matrices. However, applications to air, sludge, and soil have also been developed, so that better approaches are emerging for the analysis of all types of matrices, including gases, liquids and solids. The range of analytes that can be analyzed by SPME includes volatile, semi-volatile and even non-volatile organic and inorganic species. SPME has been successfully coupled with gas chromatography, high performance liquid chromatography, capillary electrophoresis, and mass spectrometry. Results on priority pollutants indicate that SPME can meet method requirements defined by the environmental protection agency, as performance is similar to the more traditional techniques of static headspace analysis, purge-and-trap, liquid-liquid extraction and solid phase extraction. SPME can be tuned to a given application by the choice of appropriate coating or device design. Thus far, SPME has been most broadly accepted by the food industry, particularly in the flavor and fragrance area. SPME simplifies not only the monitoring of freshness and purity of such products, but also characterization of the optimum time of harvest. In clinical applications, SPME has been used to monitor drug residues in blood, urine and other body fluids. In forensic applications, SPME has been used to monitor traces of accelerates in fire debris.
REFERENCES

1.

Janusz Pawliszyn, Solid Phase Microextraction Theory and Practice, Wiley-VCH. Inc., New York, Ny, 1997.