Solid Phase Microextraction
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1 Solid Phase Microextraction Basics, Theory and Environmental Analysis Dr. Frank Michel
2 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
3 SPME: used in thousands of applications Source ISI Search: Solid Phase Micro Extraction and SPME Published Items in Each Year Citations in Each Year Items: Total Citations: (reviews not included) H-index: 104 (reviews not included) 3
4 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 Septum-Piercing Needle Needle Ferrule Coated SPME Fused Silica Fiber
5 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
6 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! 6
7 Extraction Procedure for SPME Pierce Sample Septum Expose Fiber/Extract Retract Fiber/Remove to GC Instrument
8 Desorption Procedure for SPME Pierce GC Inlet Septum Expose Fiber/Desorb Retract Fiber/Remove to Column
9 Adsorption Mechanism for SPME Equilibrium reached Silica Rod Analyte Adsorbed Liquid Polymer Aqueous Solution Vial Extraction Time
10 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 10
11 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 e+6 ethyl benzene o-xylene toluene benzene 0.00e secs. Figure courtesy of J. Pawliszyn, et al., University of Waterloo, Ontario, Canada
12 Physical Factors Affecting Sample Recovery Influence on Equilibrium Influence on Kinetics Stiring (Temperature) A
13 Stirring in SPME Time dependence Extraction of 1,3-Dichlorobenzene Stirred Fiber coating Sample unstirred time/s Static Layer formed 13
14 Factors Affecting Sample Recovery Fiber Selection Sample Modifications Extraction Time Desorption Conditions Inlet Design Column Selection A
15 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
16 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
17 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 ,99E+03 p-nitroaniline p-nitrophenol Phenol 1,3,5- Trinitrobenzene 18
18 SPME Fibers by Adsorption Strength Estimation TPR 7µm PDMS 30µm PDMS 100µm PDMS PDMS-DVB DVB-Carboxen Carboxen Analyte Molecular Weight Range Approximation 19
19 Comparison of SPME Fibers for the Extraction of Small Hydrocarbons (Analytes at 1ppm in Air, Extracted for 10 Min) Analyte 100µm PDMS PDMS/DVB Carboxen TM /PDMS Ethane Propane Butane Pentane Hexane
20 SPME Fibers for the Extraction of Hydrocarbons n= Carboxen -PDMS Carboxen-DVB-PDMS PDMS-DVB
21 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 = at 0.6 sec/scan (quantitation ions 95 and 112) MIB Geosmin Min 22 G000169
22 SPME Technique Extraction conditions Headspace Direct Imersion Desorption temperature Liner Diameter Sample Modifications 25
23 Headspace vs. Direct Immersion Volatility of Sample Extraction Time concerns Sample Matrix Selectivity of Analytes
24 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 Min
25 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 28
26 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 Phenol Methylphenol & 4-Methylphenol Nitrophenol ,4-Dimethylphenol ,4-Dichlorophenol ,6-Dichlorophenol Chloro-3-methylphenol ,4,5-Trichlorophenol ,4,6-Trichlorophenol ,4-Dinitrophenol Nitrophenol ,3,4,6-Tetrachlorophenol Methyl-4,6-dinitrophenol Pentachlorophenol Dinoseb
27 Phenols by SPME at 50ppb (85µm Polyacrylate Fiber, ph 2) IS , Min IS 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 Chloro-3-methylphenol 11. 2,4,5-Trichlorophenol 12. 2,4,6-Trichlorophenol 13. 2,4-Dinitrophenol Nitrophenol 15. 2,3,4,6-Tetrachlorophenol Methyl-4,6- dinitrophenol IS 2,4,6-Tribromophenol 17. Pentachlorophenol 18. Dinoseb 30
28 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 31
29 Conclusion 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 32
30 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 33
31 Dziękuję za uwagę! S ample P rep M ade E asy 34
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