Solid Phase Microextraction Basics, Theory and Applications Dr. Frank Michel frank.michel@sial.com
SPME History Patented Technology of University of Waterloo, Canada Inventor: Prof. Dr. Janusz Pawliszyn Automation by Varian (AS 8200 & CTC CombiPal). CTC CombiPal makes the SPME compatibel with most GCs 2
SPME Holders For Manual Sampling (Fiber Exposed) Plunger For Varian 8100/8200 AutoSampler or SPME/HPLC Interface (Fiber Retracted) Plunger Barrel Z Slot Retaining Screw Barrel Plunger Retaining Screw Slot Hub-Viewing Window Adjustable Needle Guide/Depth Gauge Color-Coded Screw Hub Sealing Septum Fiber-Attachment Needle Retaining Nut Fiber-Attachment Needle Septum-Piercing Needle Coated SPME Fused Silica Fiber 713-0105 Septum-Piercing Needle Needle Ferrule Coated SPME Fused Silica Fiber 3 713-0107
SPME Fiber Assembly Detail (Manual) Color-Coded Screw Hub Tensioning Spring Sealing Septum Ferrule Septum-Piercing Needle Fiber-Attachment Needle Coated SPME Fused Silica Fiber 713-1344 4
Types of SPME Fiber Assemblies Assemblies for holders manual autosampler style (no spring) Gauge size of piercing needle Standard size - 24 GA Larger bore size - 23 GA (for septum free inj. ports) Types of fiber core Fused silica Stableflex Metal NEW! 5
Extraction Procedure for SPME Pierce Sample Septum Expose Fiber/Extract Retract Fiber/Remove to GC Instrument 713-1345 6 98-0369
Desorption Procedure for SPME Pierce GC Inlet Septum Expose Fiber/Desorb Retract Fiber/Remove to Column 713-1345 7 98-0370
Adsorption Mechanism for SPME Equilibrium reached Silica Rod Analyte Adsorbed Liquid Polymer Aqueous Solution Vial Extraction Time 8 98-0371
Amount of Analyte absorbed by the Fiber at Equilibrium for small sample volumes (2-5ml): n s = KV f C 0 V s KV f +V s at infinite volume of samle (V s >> V f ): n s = KV f C 0 K Distribution Costant fiber/sample n s Analyte moles into the Stationary phase V f Stationary Phase Volume V s Sample Volume Concentration of the Analyte in water C 0 9
Adsorption-time Profile for BTEX Compounds Using SPME 8.00e+6 K is compound specific Also dependend of fiber & matrix 6.00e+6 m+p-xylene Kinetics of analytes are different Higher k values require longer equilibrium times 4.00e+6 2.00e+6 ethyl benzene o-xylene toluene benzene 0.00e+0 0 100 200 300 400 500 600 700 secs. Figure courtesy of J. Pawliszyn, et al., University of Waterloo, Ontario, Canada. 10 92-0139
Physical Factors Affecting Sample Recovery Influence on Equilibrium Influence on Kinetics Stiring (Temperature) 11 95-0224A
Stirring in SPME Time dependence Extraction of 1,3-Dichlorobenzene Stirred Fiber coating Sample unstirred time/s Static Layer formed 12
Factors Affecting Sample Recovery Fiber Selection Sample Modifications Extraction Time Desorption Conditions Inlet Design Column Selection 14 95-0224A
Available SPME Fibers, by Film Type Absorption Fibers Polydimethylsiloxane (PDMS) 7, 30, and 100µm Polyacrylate (PA) Polyethyleneglycol (PEG) Adsorption fibers (with particles) Carboxen-polydimethylsiloxane(CAR-PDMS) Polydimethylsiloxane-divinylbenzene (PDMS-DVB) Divinylbenzene/Carboxen-Polydimethylsiloxane (DVB-CAR-PDMS) Unpolar Polar Polar Adsorption Adsorption Adsorption 15 98-0373
Adsorbent vs. Absorbent Fibers Adsorbent (particle) fibers Physically traps or chemically reacts bonds with analytes - porous material - high surface area Absorbent (film) fibers Analytes are extracted by partitioning liquid phase retains by thickness of coating Analytes may compete for sites Analytes do not compete for sites Fibers have limited capacity Fibers can have high capacity 16 00-0003
Area Response depending on Fiber Type 6,27E+05 Bare FS 7µm 30µm 100µm Pacrylate PDMS-DVB CW-DVB DVB-CAR Carboxen 6,80E+05 7,04E+03 4,73E+03 3,81E+03 2,58E+05 0 2,99E+03 p-nitroaniline p-nitrophenol Phenol 1,3,5- Trinitrobenzene 17
SPME Fibers by Adsorption Strength Estimation TPR 7µm PDMS 30µm PDMS 100µm PDMS PDMS-DVB DVB-Carboxen Carboxen 0 150 300 450 Analyte Molecular Weight Range Approximation 18
Odor Agents at 1ppt in Water by SPME-GC/MS Sample: 30mL water containing MIB and geosmin at 1ppt and 25% NaCl in a 40mL vial, at 65 C SPME Fiber: DVB/Carboxen /PDMS Extraction: heated headspace, 30 min, 65 C, with rapid stirring Desorption: 3 min, 250 C, splitter closed Column: Meridian MDN-5, 30m x 0.25mm x 0.25µm film Oven: 60 C (1 min) to 250 C at 15 C/min Det.: mass spectrometer, m/z = 75-180 at 0.6 sec/scan (quantitation ions 95 and 112) MIB Geosmin 6 7 8 9 Min 21 G000169
SPME Technique Extraction conditions Headspace Direct Imersion Desorption temperature Liner Diameter Sample Modifications 24
Headspace vs. Direct Immersion Volatility of Sample Extraction Time concerns Sample Matrix Selectivity of Analytes 25 95-0235
Fruit Punch Flavor by Headspace SPME Elimination of Glycerin Interference Propylene glycol Ethyl caproate Direct Injection HS SPME Glycerin Figure provided by Dr. A. Harmon, McCormick & Co., Inc., Hunt Valley, MD, USA. 6 8 10 12 Min 14 16 18 20 26
Sample Modifications Salt ph Derivatisation of analytes on the fiber Fiber saturated with reagent put into sample extraction Fiber with extracted analytes put into reagent 27
Sample Modifications The Effect of Salt and ph on Extraction of Phenols (50ppb) by SPME (PA) No Salt No Salt Salt Salt Neutral ph = 2 Neutral ph = 2 2-Chlorophenol 1800 2361 3952 14028 Phenol 810 1003 6425 6150 Methylphenol 761 882 5485 7434 3- & 4-Methylphenol 1795 1846 15337 19723 2-Nitrophenol 422 474 311 2315 2,4-Dimethylphenol 1344 1476 15000 20710 2,4-Dichlorophenol 5396 8138 19803 61664 2,6-Dichlorophenol 2991 5858 12511 48530 4-Chloro-3-methylphenol 2398 3137 24060 33529 2,4,5-Trichlorophenol 3115 11097 24270 96333 2,4,6-Trichlorophenol 9702 19307 35466 109492 2,4-Dinitrophenol 0 11 765 1182 4-Nitrophenol 626 730 11458 6536 2,3,4,6-Tetrachlorophenol 3108 27683 33938 70440 2-Methyl-4,6-dinitrophenol 55 47 920 1685 Pentachlorophenol 2305 40582 22056 143905 Dinoseb 68 2123 6676 37744 28
Phenols by SPME at 50ppb (85µm Polyacrylate Fiber, ph 2) IS 1 2 3 4,5 6 7 8 9 10 11 12 13 14 6 8 10 12 Min 14 16 18 15 16 IS 17 18 IS 2-Fluorophenol (int. std.) 1. Phenol 2. 2-Chlorophenol 3. 2-Methylphenol 4. 3-Methylphenol 5. 4-Methylphenol 6. 2-Nitrophenol 7. 2,4-Dimethylphenol 8. 2,4-Dichlorophenol 9. 2,6-Dichlorophenol 10. 4-Chloro-3-methylphenol 11. 2,4,5-Trichlorophenol 12. 2,4,6-Trichlorophenol 13. 2,4-Dinitrophenol 14. 4-Nitrophenol 15. 2,3,4,6-Tetrachlorophenol 16. 2-Methyl-4,6- dinitrophenol IS 2,4,6-Tribromophenol 17. Pentachlorophenol 18. Dinoseb 29
Quantification Internal Standard!! For complex Matrices Standard Addition Extraction is an Equilibrium!! Extraction parameter needs to be kept constant: Stir velocity Temperature Sample matrix (Salt?) Fiber position in the sample Extraction time 30
Example Applications SPME Analysis of Volatile Acids in Parmesan Cheese Limoncello Milk off-flavors Resveratrol in red wine TCA & precursors in red wine (cork taint) Honey varieties Agricultural pesticides in wine Peanut Butter Flavors by SPME Volatiles in White Wine by SPME/GC/MS Regular Coffee Grounds by SPME Peppermint oil in chocolate bar Summary 31
Analysis of Volatile Acids in Parmesan Cheese (Headspace 15 min at 65 C) 1. Acetic 2. Propionic 3. Isobutyric 4. Butyric 5. Isovaleric 6. Valeric 7. Hexanoic IS 2-Ethyl hexanoic 8. Heptanoic 9. Octanoic 10. Nonanoic 11. Decanoic 12. Undecanoic 13. Dodecanoic 4 6 7 IS SPME Fiber: 9 65µm CW-DVB 11 1 5 8 13 2 3 10 12 8 10 12 14 16 18 20 32 Min 99-0077
Conditions for Analysis of Volatile Acids in Parmesan Cheese Sample: 100mg cheese in 40mL vial SPME Fiber: 65µm CW-DVB Extraction: headspace, 15 min, 65 C Desorption: 1 min, 250 C Column: Nukol, 15m x 0.25mm, 0.25µm film Oven: 50 C (2 min) to 220 C at 10 C/min Carrier: helium, 30cm/sec Inj.: splitless/split (closed 1 min), 250 C Det.: FID, 260 C Back to Application Selection 33 Go on to Summary 99-0078
Limoncello Field: Flavors & Fragrances Sample Prep and Analysis Tools: Headspace SPME, GC Abstract: ~70 volatile compounds from limoncello captured by headspace SPME, desorbed and analyzed on SLB-5ms column. References: 34
Limoncello Limoncello 4.5 u V (x10,000) 2 3 7 12 15 17 44 48 49 55 57 Headspace SPME Analysis on SLB-5ms 4.0 3.5 3.0 2.5 5,6 11 14 19 39 45 51 2.0 10 13 30 35 1.5 1.0 0.5 1 4 8 16 18 20 24 26 47 33 43 54 52 53 59 40 58 28 31 38 42 41 60 62 63 34 0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 min Chromatogram courtesy of Prof. Luigi Mondello (Univ. of Messina, Italy) Back to Application Selection 35 Go on to Summary
Milk off-flavors Field: Flavors & Fragrances Sample Prep and Analysis Tools: SPME, GC- MS Abstract: SPME was used to detect the formation of aldehydes that are formed when unsaturated fatty acids are exposed to UV light. SPME was a useful tool for monitoring shelf life of milk. References: SPME Applications CD 36
Milk off-flavors Milk Sample Off-Flavors by SPME-GC/MS Prior to Exposure to Sunlight 1 2 3 IS SPME Fiber: 75 µm PDMS/Carboxen Sample: 3g of 2% milk + 10µL internal standard solution, (20µg/mL 4-methyl-2-pentanone) (9mL GC vial) Column: Supel-Q PLOT, 30m x 0.32mm ID Det.: GC/MS ion trap, m/z = 33-300 After 1-Hour Exposure to Sunlight 4 IS 6 1. Acetone 2. 2-Butanone 3. 3-Methylpentane 4. Pentanal 5. Dimethyldisulfide 6. Hexanal IS. 4-Methyl-2-pentanone 10 15 20 Min 5 25 30 35 Chromatogram provided by Ray Marsili, Dean Foods Technical Center, Rockford, IL, USA. Back to Application Selection 37 Go on to Summary
Resveratrol in red wine Field: Food & Beverage Sample Prep and Analysis Tools: SPME, GC (SLB-5ms), on-fiber derivatization Abstract: SPME when used in combination with on-fiber derivatization was found to be applicable to the extraction of resveratrol from red wine. The technique was found to be highly sensitive, simple, and quantitative. The polyacrylate fiber was also found to withstand exposure to the vapors of the silylating reagent without damage resulting from swelling. References: Supelco Reporter, vol. 27.4, pg. 18 38
Resveratrol in red wine Resveratrol in Merlot wine Spiked wine sample OH OTMS Extracted with SPME Analyzed by GC-MS HO OH BSTFA:TMCS TMSO MW= 444 OTMS trans-resveratrol (TMS) cis-resveratrol (TMS) 6 8 10 12 14 16 18 20 22 24 Time (min) SPME fiber: 85 µm polyacrylate Column: SLB-5ms; 30 m x 0.25 mm I.D., 0.25 µm 39
Resveratrol in red wine Linearity of Method Trans-resveratrol standards prepared in 12% ethanol in water 3500000 SPME - on fiber derivatization analysis of trans resveratrol peak area 3000000 2500000 2000000 1500000 1000000 500000 0 R 2 = 0.9987 0 50 100 150 200 250 300 350 Conc. (ug/l) Quantification of spiked wine sample Unspiked red wine Spiked red wine (100 ug/l) Conc. of trans-resveratrol (ug/l) 22.6 134.7 % recovery --- 110% 40
Resveratrol in red wine SPME-GC/MS Analysis of red wine samples trans-resveratrol (TMS) cis-resveratrol (TMS) unspiked wine 6 8 10 12 14 16 18 20 22 24 Time (min) cis-resveratrol (TMS) trans-resveratrol (TMS) spiked wine 6 8 10 12 14 16 18 20 22 24 Time (min) Back to Application Selection 41 Go on to Summary
TCA & precursors in red wine (cork taint) Field: Food & Beverage Sample Prep and Analysis Tools: Headspace SPME, GC (SLB-5ms) Abstract: The ability of headspace SPME, in combination with analysis on the SLB-5ms, to detect a low level of 2,4,6- trichloroanisole (TCA) in wine is shown here. TCA is often the source of the musty smell in wine resulting from tainted corks. This is another example of how the SLB- 5ms column provides low bleed and inertness to meet the demands of today s sensitive GC-MS and GC applications. References: Supelco Reporter, vol. 24.4, pg. 14 42
TCA & precursors in red wine (cork taint) TCA & precursors in red wine (cork taint) TCA 1 2 3 4 4 6 8 10 12 Time (min) 43
TCA & precursors in red wine (cork taint) SPME of Cork Taint and its precursors Cork taint or a musty odor sometimes detected in wine, is the result of 2,4,6- trichloroanisole (TCA). The source of TCA is thought to be the fungal methylation of chorophenols present in the wine, with these compounds emanating from the cork or other sources such as biocides, fungicides, and exposure of processing equipment to antiseptic cleaning products containing halophenols (1). This application demonstrates the use of solid phase microextraction (SPME) for the analysis of TCA and several chlorophenolic precursors from wine. The chlorophenols were derivatized in matrix using acetic anhydride, and the acylated derivatives extracted from the headspace. The TCA, which is not derivatized, was simultaneously extracted with the halophenols. Final analysis was performed by GC-ECD on the SLB-5ms capillary column. Target analytes were: 2,4,6-Trichloroanisole 2,4,6-Trichlorophenol 2,3,4,6-Tetrachlorophenol Pentachlorophenol 44
TCA & precursors in red wine (cork taint) Extraction and analysis conditions sample: 1.5 ml sample + 600 µl 5% K2CO3 + 240 mg NaCl + 60 µl acetic anhydride SPME fiber: metal fiber coated with 100 µm PDMS (57928-U) extraction: desorption temp.: column: headspace, 50 C, 30 min., with stirring 250 C, 3 min. SLB-5ms; 30 m x 0.25 mm I.D. x 0.25 µm (28471-U) oven: 50 C (1 min.), 25 C/min. to 280 C detector: ECD, 290 C carrier gas: liner: 0.75 mm I.D. SPME helium, 1.5 ml/min constant flow 45
TCA & precursors in red wine (cork taint) Derivatization reaction The halophenols were acylated with acetic anhydride prior to extraction. Acetic anhydride will hydrolyze in the presence of water, however the phenolic groups present on the analytes are more reactive, making it possible to conduct derivatization in an aqueous matrix (2). The addition of K 2 CO 3 drives the reaction by removing the acetic acid that is formed: O C H 3 O O CH 3 + Cl OH Cl Cl O CH 3 Cl + H 3 C O OH O Cl Cl 46
TCA & precursors in red wine (cork taint) Linearity Standards from 10-300 ng/l prepared in 12% ethanol in water were derivatized, extracted, and the response vs. concentration was plotted: Area counts 25000 20000 15000 10000 2,4,6-TCA 2,4,6-TCP 2,3,4,6-TeCP PCP Linear (PCP) SPME-Headspace Extraction R 2 = 0.9747 R 2 = 0.9817 R 2 = 0.9864 R 2 = 0.9957 Standards in 12% ethanol in water 5000 0 0 50 100 150 200 250 300 Conc. (ng/l) Good linearity was obtained, indicating the method to be quantitative 47
TCA & precursors in red wine (cork taint) SPME-GC/ECD Analysis of wine sample The wine sample used for extraction was a California shiraz in a wax-lined carton-type container with a plastic closure. This type of packaging was chosen to minimize the presence of cork taint compounds. unspiked wine 2 4 4 6 8 10 12 Time (min) spiked wine 1 2 3 4 1. 2,4,6-Trichloroanisole 2. 2,4,6-Trichlorophenol (acylated) 3. 2,3,4,6-Tetrachorophenol (acylated) 4. Pentachlorophenol (acylated) 4 6 8 10 12 Time (min) 48
TCA & precursors in red wine (cork taint) Quantification of spiked wine sample Unspiked and spiked (100 ng/l) red wine samples were extracted and recovery was determined against the calibration curves generated using the extracted standards in 12% ethanol. Spike level of 100 ng/l Unspiked wine (ng/l) Spiked wine (ng/l) % Rec. 2,4,6-Trichloroanisole ND 60.7 61 2,4,6-Trichlorophenol 22.7 96.3 74 2,3,4,6-Tetrachlorophenol ND 55.7 56 Pentachlorophenol 3.3 33.5 30 All four analytes were recovered, however it appears that the wine matrix may have interfered with accuracy to some extent, as indicated by the % recovery values. 49
TCA & precursors in red wine (cork taint) Reproducibility A check of reproducibility was performed by doing extractions of sets of three spikes prepared in the 12% ethanol in water and red wine. (area counts) 2,4,6-TCA 2,4,6-TCP 2,3,4,6-TeCP PCP 12% EtOH 100 4272 4495 2775 5011 12% EtOH 100 4063 4374 3074 5766 12% EtOH 100 4235 4299 3168 6055 Avg; 12% EtOH 4190 4389 3006 5611 std. Dev 112 99 205 539 % RSD, 12% ETOH 3% 2% 7% 10% wine 100 3298 4231 1977 2750 wine 100 2986 3916 1613 2335 wine 100 2727 2341 1843 2745 Avg.; w ine 3004 3496 1811 2610 std. Dev 286 1013 184 238 % RSD, spiked wine 10% 29% 10% 9% Reproducibility for the 12% ethanol in water samples was good, with %RSD values of <10%. The data indicates the effect of matrix, with average area counts lower and more variable overall for the wine samples. 50
TCA & precursors in red wine (cork taint) Conclusions SPME can be used for the extraction of 2,4,6-trichloroanisole and its halophenolic precursors from wine. Derivatization makes the analytes easier to extract and analyze by GC. Headspace extraction in combination with ECD can be used to reduce background interference. The method appears to be quantitative, although further work would be necessary to optimize extraction efficiency from wine matrix. 51
TCA & precursors in red wine (cork taint) References 1. Insa, S., Salvado, V., Antico, E., Development of solid-phase extraction and solid-phase microextraction methods for the determination of chlorophenols in cork macerate and wine samples. J. Chromatogr. A, 2004, 1047: 15-20. 2. K. Blau; J. Halket, Handbook of Derivatives for Chromatography, Second Edition, John Wiley & Sons, New York, 1993. pp 38. Back to Application Selection 52 Go on to Summary
Honey varieties Field: Food & Beverage Sample Prep and Analysis Tools: SPME, GC Abstract: In this work we evaluate the applicability of headspace SPME (HS-SPME) coupled with gas chromatography-mass spectrometry (GC- MS) for the characterization of the volatile fraction of some honey samples. References: Supelco Reporter, vol. 28.1, pg. 3 53
Honey varieties Milk Thistle Honey Volatiles Using SPME on the SUPELCOWAX 10 6 7 9 5 10 1 11 0 10 20 30 Time (min) Chromatogram courtesy of Dr. Federica Bianchi and Prof. Marilena Musci (Univ. of Parma, Italy) 54
Honey varieties Citrus Honey Volatiles Using SPME on the SUPELCOWAX 10 6 7 3 10 0 10 20 30 Time (min) Chromatogram courtesy of Dr. Federica Bianchi and Prof. Marilena Musci (Univ. of Parma, Italy) 55
Honey varieties Eucalyptus Honey Volatiles Using SPME on the SUPELCOWAX 10 1 8 11 0 10 20 30 Time (min) Chromatogram courtesy of Dr. Federica Bianchi and Prof. Marilena Musci (Univ. of Parma, Italy) 56
Honey varieties Acacia Honey Volatiles Using SPME on the SUPELCOWAX 10 4 5 1 6 10 0 10 20 30 Time (min) Chromatogram courtesy of Dr. Federica Bianchi and Prof. Marilena Musci (Univ. of Parma, Italy) 57
Honey varieties Multifloral Honey Volatiles Using SPME on the SUPELCOWAX 10 6 1 4 8 2 11 0 10 20 30 Time (min) Chromatogram courtesy of Dr. Federica Bianchi and Prof. Marilena Musci (Univ. of Parma, Italy) Back to Application Selection 58 Go on to Summary
Agricultural pesticides in wine Field: Food Safety Sample Prep and Analysis Tools: SPME, GC Abstract: This application demonstrates the usefulness of SPME in the low-level extraction of agricultural pesticides from wine, and the use of the SLB-5ms in the subsequent analysis. The pesticides chosen for the analysis represent a group of insecticides and fungicides that could be found in commercial wines. These compounds contain a variety of polar functional groups, and the polyacrylate fiber provided the selectivity necessary for extraction from a wine matrix. The inertness and low bleed of the SLB-5ms enabled subsequent low-level analysis of these compounds by GC-MS. References: Supelco Reporter, vol. 24.4, pg. 14 59
Agricultural pesticides in wine Agricultural pesticides in wine SPME-GC/MS Analysis of spiked wine sample Peak IDs: 1. Dicloran 2. Diazinon 3. Chloropyrifos-methyl 4. Vinclozolin 5. Carbaryl 6. Methiocarb 3 7. Dichlofluanid 8. Parathion-ethyl 9. Triadimefon 10. Procymidone 11. Myclobutanil 12. Imidan (Phosmet) 13. Dicofol 14. Phosalone 15. Azinphos-methyl 1 2 4 5 6 7 8 9 column: SLB-5ms, 30 m x 0.25 mm I.D., 0.25 µm (28471-U) SPME fiber: 85 µm polyacrylate (57304) extraction: immersion, room temp. (30 min.) desorption: 5 min. at 250 C oven: 60 C (1 min.), 15 C/min to 100 C, 7 C/min. to 300 C (1 min.) MSD interface: 325 C scan range: SIM carrier gas: helium, 0.7 ml/min., constant liner: 0.75 mm I.D. SPME liner sample: white wine spiked with 50 ppb pesticides 10 11 Unk. 12 13 14 15 18 20 22 24 26 28 30 Time (min) Back to Application Selection 60 Go on to Summary
Peanut Butter Flavors by SPME Sample: 5g peanut butter in 40mL vial SPME Fiber: DVB-Carboxen -PDMS (StableFlex Fiber) Extraction: headspace, 30 min at 65 C in heating block Desorption: 5 min, 270 C 9 10 15 16 19 Column: SUPELCOWAX 10, 30m x 0.25mm x 0.25µm film Oven: 40 C (5 min) to 230 C at 4 C/min Inj.: splitless/split, closed 0.5 min, 270 C, with 0.75mm liner Det.: ion trap mass spectrometer, m/z = 30-350 at 0.6 sec/scan Selected ions used for quantitation. 1 2 7 11 12 14 21 22 28 3 4 5 6 8 13 20 17 18 26 24 25 27 29 0 6 12 Min 18 23 24 G000517 61 98-0394
Flavor Components in Peanut Butter Some Volatile Components in Peanut Butter 1. Carbon disulfide 2. 3-Methylbutanal 3. Pentanal 4. Dimethyl disulfide 5. Hexanal 6. 4-Methyl-pentene-2-one 7. 1-Methyl pyrrole 8. Heptanal Pyrazines in Peanut Butter 9. 2-Methyl pyrazine 10. 2,5-Dimethyl pyrazine 11. 2,3-Dimethyl pyrazine 12. 2-Ethyl pyrazine 13. 2,6-Dimethyl pyrazine 14. 2-Ethyl-6-methyl pyrazine Pyrazines in Peanut Butter (contd.) 15. 2-Ethyl-5-methyl pyrazine 16. Trimethyl pyrazine 17. 2-Ethyl-3-methyl pyrazine 18. 2,6-Diethyl pyrazine 19. 2-Ethyl-3,5-dimethyl pyrazine 20. 2,3-Diethyl pyrazine 21. 2-Methyl-5-isopropyl pyrazine 22. 3-Ethyl-2,5-dimethyl pyrazine 23. 5-Methyl-2-propyl pyrazine 24. 2-Methyl-5-propyl pyrazine 25. 2-Ethenyl-6-methyl pyrazine 26. 3,5-Diethyl-2-methyl pyrazine 27. 2-Ethenyl-5-methyl pyrazine 28. 2-Methyl-6-cis propenyl pyrazine 29. 2-Allyl-5-methyl pyrazine Back to Application Selection 62 Go on to Summary 98-0395
Volatiles in White Wine by SPME/GC/MS Sample: White wine + 25% NaCl SPME Fiber: Carboxen /PDMS Extraction: headspace, 10 min, 40 C Desorption: 3 min at 290 C Column: VOCOL, 30m x 0.25mm ID, 1.5µm film Detector: GC/MS, quadrupole, m/z = 31-240 2 5 7 8 12 11 13 16 1. Sulfur dioxide 2. Ethanol 3. Methyl formate 4. Acetic acid 5. Ethyl acetate 6. Isobutanol 7. Isopentanol 8. 2-Methyl-1-butanol 9. Ethyl butyrate 10. 2,3-Butanediol 11. Hexanol 12. Isoamyl acetate 13. Ethyl hexanoate 14. Hexyl acetate 15. Octanoic acid 16. Ethyl octanoate 1 3 4 6 9 10 14 15 98-0377 5 10 Min 15 20 Back to Application Selection 63 Go on to Summary
Regular Coffee Grounds by SPME Sample: 5g coffee grounds in 40mL vial SPME Fiber: DVB/Carboxen /PDMS (StableFlex Fiber) Extraction: headspace, 30 min at 65 C Desorption: 270 C for 5 min Column: SUPELCOWAX 10, 30m x 0.25mm x 0.25µm film Oven: 40 C (5 min) to 230 C at 4 C/min Inj.: splitless/split, closed 0.5 min, 270 C, with 0.75mm liner Det.: ion trap mass spectrometer, m/z = 30-350 at 0.6 sec/scan Selected ions used for quantitation. 1 3 4 2 5 6 15 14 7 8 910 11 1213 16 17 23 22 32 24 26 21 27 28 29 25 31 19 18 20 30 36 42 33 34 39 40 35 38 37 41 43 46 44 45 47 48 49 51 50 52 53 54 55 56 57 4 8 12 16 20 Min 24 28 32 36 40 64 98-0397
Components in Coffee 1. 2-Methyl furan 2. 2-Butanone 3. 2-Pentanone 4. 3-Methyl butanal 5. 2,5-Dimethylfuran 6. 2-Acetyloxy-2-propanone 7. 2-Ethyl hexanol 8. Dimethyldisulfide 9. Phenol 10. Hexanal 11. 2-Methyl thiophene 12. n-methyl pyrrole 13. 4-Methylphenol 14. 2-Ethyl pyrrole 15. Pyridine 16. Pyrazine 17. Methyl pyrazine 18. 4-Methyl thiazole 19. 3-Hydroxy butanone 20. Dimethyl phenol (isomer) 21. 1,2-Ethanediol, monoacetate 22. 2,5-Dimethylpyrazine 23. 2,3-Dimethylpyrazine 24. 2-Ethylpyrazine 25. 2,6-Dimethylpyrazine 26. 2-Ethyl-6-methylpyrazine 27. 2-Ethyl-5-methylpyrazine 28. Trimethylpyrazine 29. 2-Ethyl-3-methylpyrazine 30. 2,6-Diethylpyrazine 31. 2-Ethenylpyrazine 32. 2-Ethyl-3,5-dimethylpyrazine 33. Glycerol 34. 2,3-Diethylpyrazine 35. 2-Ethyl-3,6-dimethylpyrazine 36. 2-Furancarboxaldehyde 37. 2-Isopropenylpyrazine 38. 3,5-Diethyl-2-methylpyrazine 39. Furfural formate 40. 2-Furonyl ethanone 41. Methyl benzoylformate 42. Furanmethanol acetate 43. 5-Methyl-2-furancarboxaldehyde 44. Furanmethanol proprionate 45. Furfanyl furan 46. Pyridine methanol 47. 2-Methyl-5-propenylpyrazine 48. Furanmethanol 49. 3-Ethyl-4-methyl-2,5-furandione 50. Pyrazinecarboxamide 51. 2-Ethyl-3-hydroxy-4H pyran-4-one 52. 1-(2-Furanylmethyl)-pyrrole 53. 2-Methoxyphenol 54. 1-(1H-pyrrole-2-yl)-ethanone 55. 4-Ethyl-2-methoxy phenol 56. 3-Phenylpropenal or 2-Methylbenzofuran 57. 3,5-Dimethylbenzoic acid Back to Application Selection 65 Go on to Summary 98-0401
Peppermint oil in chocolate bar 2 4 Sample: 4g peppermint cookie bar SPME Fiber: 100µm PDMS Extraction: headspace, 1 min, 45 C Desorption: 5 min at 250 C Column: PTE -5, 30m x 0.25mm ID, 0.25µm film Detector: FID, 250 C Injector: splitless (3 min), 250 C 1 3 1. Internal standard 2. cis-menthone 3. trans-menthone 4. Menthol Min 96-0116 0 4 8 12 Back to Application Selection 66 Go on to Summary
Official methods for SPME ISO Standard 27108 (derived from DIN 38407-F34) Determination of selected plant treatment agents and biocide products - Method using solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) ISO Standard 17943 (derived from DIN 38407-41) Water quality - Determination of VOCs in water - Method using HS- SPME followed by GC-MS OENORM A 1117, 2004-05-01 Determination of volatile compounds in cellulose-based materials by Solid Phase Micro Extraction (SPME) EPA Method 8272 (Dec 2007) Parent and Alkyl Polycyclic Aromatics in Sediment Pore Water by SPME GC/MS 67
Official methods for SPME ASTM D 6438, 2005 Standard Test Method for Acetone, Methyl Acetate, and Parachlorobenzotrifluoride Content of Paints, and Coatings by SPME/GC ASTM D 6520, 2000 Standard Practice for the SPME of Water and its Headspace for the Analysis of Volatile and Semi-Volatile Organic Compounds ASTM D 6889, 2003 Standard Practice for Fast Screening for Volatile Organic Compounds in Water Using SPME ASTM E 2154, 2001 Standard Practice for Separation and Concentration of Ignitable Liquid Residues from Fire Debris Samples by Passive Headspace Concentration with SPME 68
Summary One Step Extraction Micro Technology Trace Analysis 100% Solvent Free Equilibrium Technology Control your T s (Time, Temp., Technique) Suitable for Liquids (Water), Gases or Solids Quantitative Automation possible CTC Gerstel Varian Thermo 69
Supelco SPME Bulletins # 925 SPME-Applications Guide (only on CD & web) # 923 Theory and Optimization of Conditions # 928 Trouble Shooting guide # 929 Practical Guide to Quantification SPME # 901 Drugs, Alcohol, org. Solvents in Biological Fluids # 922 Forensic Applications: Explosives, Fire Debris, and Drugs of Abuse # 869 Flavour and Fragances All on the SPME-CD plus additional applications & videos 70
Thank you! S ample P rep M ade E asy 71