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Order](https://www.sigmaaldrich.com/JP/en/quick-order) Cart0 [Home](https://www.sigmaaldrich.com/US/en)[Solid Phase Microextraction (SPME)](https://www.sigmaaldrich.com/US/en/applications/analytical-chemistry/sample-preparation/solid-phase-microextraction)Solid Phase Microextraction: Frequently Asked Questions # Solid Phase Microextraction: Frequently Asked Questions ### Q: What SPME fiber should I use to extract a class of analytes? Two factors should be considered: - Polarity of the analyte - Volatility and molecular size of the analyte Polar analytes are attracted to polar phases (e.g., polyacrylate and Carbowax coatings). Fibers with a thicker film (100µm) are better for volatiles, but also can be used for less volatile compounds (longer extraction times are required). Porous fibers (with Carboxen or divinylbenzene coating) also can retain small analytes, and are ideal for C2-C6 analytes Thinner film fibers (7µm and 30µm polydimethylsiloxane) are better for larger molecules. For more information, SPME Fiber Selection. ### Q: How can I improve SPME recovery? - Select the most appropriate fiber (see the previous question). - Agitate or stir the sample. - If you are performing headspace sampling, minimize the headspace volume (<30%) Select the appropriate temperature (40-90 °C). Analytes can desorb from the fiber if the temperature is too high. - Add 25% NaCl to the sample and adjust the pH (reduce the pH for acids, increase the pH for bases). - Minimize the amount of organic solvent in the sample. ### Q: Is SPME quantitative? Yes. A calibration curve to determine linear range is required for each analyte. Using internal standards with properties similar to those of the analytes in the sample improves precision. Usee of Standard Additions is best used for complex matrices. For gas chromatography/mass spectrometry analysis, isotopes are ideal as internal standards. Consistent extraction conditions and stirring or agitation is required. ### Q: How do I quantify results from manual SPME? For reliable quantitative results from manual SPME extractions, we recommend these guidelines: The sample volume must be consistent among samples. For direct immersion applications, completely fill the sample container. For headspace applications, fill the container between 1/2 and 3/4 full. When extracting semivolatiles, use a consistent stirring rate. If the rate is inconsistent, precision drops, particularly for analytes with high distribution constants. Keep the amount of organic content in the samples minimal (<1%) and consistent among all samples. Large amounts of organic solvent can greatly affect the distribution constants. Some solvents, such as methanol, act as co-solvents with the water, and can reduce the recovery of analytes extracted. Use internal standards that are similar to your analytes. Internal standards help to offset extraction variables. Standard preparation is important. Prepare standards at 1-2mg/mL solvent, then spike 0.05-5uL of the standard into the water sample prior to the extraction. Keep extraction time and temperature consistent among samples. The extraction time generally is 10 minutes for volatiles, 20-30 minutes for semivolatiles. For more information about consistent results from SPME, refer to [Bulletin 923](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/119/860/milliflex-quantum-services-ds1649en-ms.pdf). ### Q: How can I extend SPME fiber life? Most fiber damage is the result of needle bending. Bending usually occurs while the fiber is piercing the vial septa. Adjust the black needle depth gauge so that only 0.5-1cm of the needle is exposed. Pierce the vial septa, then screw the needle into the vial by rotating the needle depth gauge. Maintain the injection port temperature below the maximum recommended temperature of the fiber. Conduct headspace sampling when ever possible to minimize extraction or carry over of high molecular weight compounds that coat the fiber. ### Q: My background is too noisy. How can I reduce extraneous peaks? - Proper conditioning of the fiber before using it. Recondition the fiber for a longer time. - Check the inlet liners. Most peak noise is caused by septa particles in the liner. - Change the vial septa. - Reduce the inlet temperature by about 20 °C. Use the lowest temperature that produces sharp peaks and minimizes carryover. ### Q: Can analytes be extracted from organic solvent using SPME? We do not recommend extraction from organic solvents. However, you may extract analytes using the headspace above polar organic solvents, at the risk of a reduced distribution constant, fiber damage from swelling, and a large solvent peak on your chromatogram. ### Q: What are the advantages of headspace SPME versus Headspace Analyzers? In analyses of semivolatile and volatile analytes, headspace SPME offers several important advantages over headspace analyzers. When you use a headspace analyzer to trap semivolatile analytes in water-based samples, you must heat the sample to 110 °C or higher. Much water will be transferred with the analytes. Often the analyte-containing volume is large and you must cryogenically refocus the analytes at the column inlet. Water that is transferred with the analytes can freeze in the line. With headspace SPME, samples usually require warming only to 40-60 °C. The analytes will concentrate on the fiber with little water vapor. Then, because the fiber is desorbed rapidly in the injection port, the analytes usually desorb as a focused plug -- no cryogenics required. In addition, with headspace SPME sample volume is small, e.g., 3 mL in a 4 mL vial. Even volatile organic compounds (VOCs) can be desorbed from an SPME fiber with sufficient speed to eliminate the need for cryogenics. Use a Carboxen™/polydimethylsiloxane fiber to retain extremely volatile analytes (gases at room temperature). Only ethane, methane, and some fixed gases are too small to be retained in the pores of Carboxen particles. Other SPME fibers are suitable for extracting semivolatiles by headspace SPME. The decision to use headspace SPME or fiber immersion SPME is based almost entirely on the vapor pressure of the analytes. If there is significant vapor pressure, headspace SPME is advantageous; analyte recoveries are cleaner and faster than by immersion SPME, and the fiber will last longer. If you have a solid sample matrix, headspace SPME is mandatory. __Related Articles__ - [Determination of Formaldehyde and Acetaldehyde in Air Using DNPH Cartridges and Automated On-Line Desorption Followed by HPLC](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/environmental-testing-and-industrial-hygiene/air-testing/aldehydes-air-dnph) - [The Effect of Container Materials on Production of Light Induced Off-Flavors in Milk – A Study Using SPME-GC-MS](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/gas-chromatography/container-materials) - [Analysis of Pesticide Residues in Ginger Powder Using QuEChERS Extraction and Clean-up with a Novel Dual Layer SPE Cartridge](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/quechers/analysis-of-pesticide-residues-in-ginger-powder) - [Analysis of Pesticide Residues in Pistachios Using QuEChERS Extraction and Cleanup with Supel™ QuE Z-Sep+](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/quechers/analysis-of-pesticide-residues-in-pistachios) - [HybridSPE-Phospholipid Technology](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/small-molecule-hplc/hybridspe-precipitation-technology) - [Impact of Mobile Phase Additives on LC-MS Sensitivity, Demonstrated using Spice Cannabinoids](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/small-molecule-hplc/lcms-spice-cannabinoids) - [Analysis of Drugs of Abuse in Urine After Cleanup with HLB Solid Phase Extraction 96-well Plates](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/solid-phase-extraction/analysis-of-drugs-of-abuse-in-urine-after-cleanup-with-hlb) - [Chiral and Achiral LC-MS Analysis of Warfarin™ in Plasma Samples](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/solid-phase-extraction/chiral-and-achiral-lc-ms-analysis-of-warfarin-in-plasma-samples) - [View More](https://www.sigmaaldrich.com/US/en/search/facet-search?focus=sitecontent&term=facet-search) Top __Sign In To Continue__ To continue reading please sign in or create an account. 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[Small Molecule HPLC](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/liquid-chromatography/small-molecule-hplc) - [Mass Spectrometry (MS)](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/mass-spectrometry) - [Nuclear Magnetic Resonance (NMR)](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/nuclear-magnetic-resonance) - [Photometry & Reflectometry](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/photometry-and-reflectometry) - [Sample Preparation](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/sample-preparation) - [Sample Filtration](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/sample-preparation/filtration) - [Sample Purification](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/sample-preparation/purification) - [QuEChERS Sample Preparation](https://www.sigmaaldrich.com/JP/en/applications/analytical-chemistry/sample-preparation/quechers) - 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