Recent Developments in SPME - Sigma-Aldrich · Recent Developments in SPME Metal Fibers Coating Process Official Methods. sigma-aldrich.com 2 Content • Metal SPME Fibers ... extraction

Recent Developments in SPME

Metal FibersCoating ProcessOfficial Methods

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Content

• Metal SPME Fibers• New PEG Fiber• DIN Method for Pesticides in Water with SPME

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SPME Fiber Assembly Detail (Manual)

713-1344

94-0034

Color-Coded Screw Hub

Tensioning Spring

Sealing Septum

Ferrule

Septum-Piercing Needle

Fiber-Attachment Needle

Coated SPME Fused Silica Fiber

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CTC Analytics Combi Pal

License bearer:CTC, Varian, Gerstel, Thermo, Alfatech, HTA, PAS, Leap

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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!

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Metal Alloy SPME Fibers

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Why Develop New SPME Metal Fiber Assemblies?

• Extend assembly lifetime

• Improve the durability and reproducibility of the extraction fiber

• Enhance compatibility with a autosampler (e.g. CTC)

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Super Elastic vs. Standard SPME Fiber Assemblies

Needle

Plunger

Metal SPME Fiber

Standard SPME Fiber

Super Elastic SPME Fibers

Standard SPME Fibers

Needle 23 gauge metal alloyBeveled tip

24 or 23 gauge. SS

Blunt tip

Fused silicaor Stableflex

SS tubing

Fiber Core Metal alloy

Plunger Solid metal alloy

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Advantages Of The New Assembly Design

• Increased durability results in up to 10 times the fiber life • Inert fiber core, does not contain iron• Coating bonds better to the metal core compared to the

Stableflex core• Crimped in assembly without adhesives.

- Does not produce extraneous peaks

• Stronger, more flexible plunger• Beveled needle tip for easier septa penetration

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Superelasticity

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Comparison of Fiber Cores (PDMS-DVB)

0 2 4 6 8Time (min)

1

2

3

4 5

Fused SilicaStableflex

Metal

1. Methylamine2. Dimethylamine3. n-Propanol (IS)4. Diethylamine5. Triethylamine

Sample: 1ppm each in waterwith 25%NaCl, 0.05M phospateBuffer pH 11

Extraction: 10min immersionFiber: PDMS-DVB various coreDesorb: 3min @ 250°CColumn: 30m x 0.32mm x 4.0µm

SPB-1 Sulfur (amine deactivate)

Oven: 45°C(1.5min) to 80°C@ 8°C/min to 200°C@

20°C/minInjector: 250°C, splitless closed

initial 0.75min. Detector: FID

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Improvements for Metal Fibers

• Advanced Metal Working Techniques - Precision EDM cutting and drilling- Specially designed crimp tool with torque measurement- Additional cleaning of alloy parts during processing

• Quality Assurance Improvements- QA data sheet for individual assemblies- 100% Inspection of critical parameters- Individual lot number assignments for assembly tracking- Raw material tracking

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Improvement of Fiber Coating

• Up to now - Manual process- Fiber strands of approx. 80-90

cm

• New coating technique- Coating tower- in-line curing- Fiber strands of 20m possible

→ More reproducible coatings→ More reproducible results

Coating &

Curing

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Variation Between Batch-Coated Lots

Analytes Average % RSD Analytes Average % RSDMethanol 217 24,3% 1-Butanol 2155 22,8%DMA 7223 43,4% TEA 4529 8,0%Ethanol 535 16,8% Dioxane 1694 23,3%Acetonitrile 1149 21,5% Pyridine 1772 20,4%Acetone 2052 23,2% 2-Hexanone 3227 16,9%Isopropano 1475 20,8% Hexanal 215 12,6%n-Propanol 1412 21,2% Aniline 4234 22,2%MTBE 1531 23,7% 21,1%

16,0%Overall Average

Coating Average, 59µm

(PDMS-DVB n = 36, 6 fibers per lot, 6 lots)

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Variation Between Automated-Coated Lots

Analytes Average %RSD Analytes Average %RSDMethanol 291 7.1% 1-Butanol 3119 9.6%DMA 5069 19.1% TEA 4747 8.5%Ethanol 715 9.1% Dioxane 2405 8.6%Acetonitrile 1571 7.8% Pyridine 2599 8.5%Acetone 2873 6.9% 2-Hexanone 3939 3.3%Isopropanol 2118 9.9% Hexanal 242 7.8%n-Propanol 2011 9.8% Aniline 4745 5.5%MTBE 2126 7.4% Overall 8.6%

Coating µm 71.7 4.1%

(PDMS-DVB n = 36, 6 fibers per lot, 6 lots)

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Important Notes About Metal Assemblies

• Superelastic assemblies will core septa, they should be used with Merlin Microseal or other septum-less injection port heads

• Adjust CombiPal home position to insure fiber does not extend beyond the needle during microseal penetration.- Merlin Microseal can be cut if not adjusted

• The assembly can be damaged by excessive heat or conditioning time.

• Use the appropriate sample vial and tops

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New Instructions for Set Up and Use of the Super Elastic Assembly

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Available Metal Fibers

• Particle Phases- 57902-U 65 µm PDMS/DVB, Metall Alloy, 1 cm length, - 57906-U 85 µm Carboxen/PDMS, Metall Alloy, 1 cm length- 57912-U 50/30 µm DVB/Carboxen-PDMS, Metall Alloy, 1 cm length- 57914-U 50/30 µm DVB/Carboxen-PDMS, Metall Alloy, 2 cm length

• Film Phases- 57919-U 7 μm PDMS, Metall Alloy, 1 cm length- 57922-U 30 μm PDMS, Metall Alloy, 1 cm length- 57928-U 100 μm PDMS, Metall Alloy, 1 cm length

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New Polyethylene glycol (PEG) SPME Fiber

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PEG SPME FiberGoals for Development of New Polar Fiber

• Find a replacement for discontinued Carbowax/DVB fiber

• Develop a polar fiber with a durable coating that will withstand multiple extractions from water

• Develop a polar fiber that does not contain an adsorbent• Put the coating on an unbreakable metal fiber core• Compare new coating with other fiber coatings• Develop a fiber that enhances selectivity for polar

analytes

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Extraction of Solvents in Water with 60µm PEG Fiber

min0 2 4 6 8 10 12

uV

600

800

1000

1200

1400

1600

1

2

3

4

5

6

7

89

10

1112

Component ppm 1. Methanol 202. Ethanol 20 3. Acetonitrile 204. Acetone 205. Isopropanol 20 6. n-Propanol 20

Component ppm 7. MTBE 38. Ethyl acetate 59. n-Butanol 10

10. 1,4-Dioxane 2011. Butyric acid 512. Phenol 0.2

Sample Polar solvents in water with 25%NaCl Fiber: 60µm PEGExtraction: 10min, Direct Immersion with agitation (Varian 8200)Desorption: 5min at 240°CInjector: Splitless/split, closed initial 0.75min than opened 50:1

with 0.75mm ID linerColumn: SPB-1 Sulfur, 30m x 0.32mm, 4.0µmColumn oven: 45°C(1.5min) to 80°C at 8°C/min to 230°C at 20°C/min

(10min)Carrier gas: He @t 13psi const. pressure, 40cm/sec at 40°CDetector: FID at 300°C

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Comparison of Fibers for Extraction of Solvents

22375

0

3000

6000

9000

12000

15000

Methanol Ethanol Acetonitrile Acetone IPA n-PrOH MTBE Et Acetate n-BuOH 1,4,Dioxane ButryicAcid

Phenol

Are

a co

utns

85µm PA60µm PEGCW/DVB 100um PDMS

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Durability of PEG Fibers with Repeated Extractions

0

100

200

300

400

500

600

700

0 15 30 45 60 75 90 105

Extraction number

Are

a co

unts

Ethanol 20ppmAcetonitrile 20ppmAcetone 10ppmIsopropanol 10ppmn-Propanol 10ppm1,4,Dioxane 20ppm

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Comparison of Fibers, Phenols 25ppb

1987222

0

50000

00000

50000

00000

50000

00000

Pheno

l2-C

l-Ph

3-MeP

hUnd

ecane

2-Nitro

P2,4

-DMP

2,4-D

ClPNap

hthalen

e4-C

l-3-m

eP2,4

,5-TClP

2,4-di

nitroP

4-nitro

P2,3

,4,6-t

eClP

2-me-4

,6-diN

P

PClPDino

seb

PAPEGCWDVB

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Comparison of SPME Fibers Base-Neutrals (Basic Fraction)

3777

8

2088

5

1556

58

3249

48

2088

06

8987

2

6461

1

5741

0

3316

9

1860

25

5341

94

3906

94

2517

79 3104

21

1859

3

3558

8725

2

2449

29

1438

93

6442

6

7261

2

4217

405

5745

3

1982

16

1856

806

1202

0

100000

200000

300000

400000

500000

600000

Aniline Benzyl alcohol o-Toluidine p-Chloroaniline o-Nitroaniline m-Nitroaniline p-Nitroaniline

85µm Polyacrylate60µm PEG65µm CW-DVB100µm PDMS

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Comparison of SPME Fibers Base-Neutrals (Neutral Fraction)

0,0E+00

2,0E+06

4,0E+06

6,0E+06

8,0E+06

1,0E+07

1,2E+07

1,4E+07

1,6E+07

1,8E+07

Undecane Benzoanthracene Naphthalene Acenaphthene

Are

a co

unts

85um PA60µm PEGCW/DVB100um PDMS

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Applications with SPME PEG FiberOrganophosphorus Pesticides at 10ppb by SPME

Extraction:

Sample: OPPs at 10ppb in 0.1M Phosphate buffer, pH 7, 25% NaCl30min, direct immersion with agitation (CombiPal)

Fiber: 60µm PEGDesorption: 2min at 250°C, splitless for 0.75min than opened Column: SLB-5 MS 30m x 0.25mm ID, 0.5µm film Oven: 60°C(1min) to 215°C @20°C/min to 290°C

@ 6°C/min (1min)Carrier: Helium, 1mL/min constant flow, Injector: Splitless/split w/ 0.75mmID linerDetector ITMS m/z = 50-326 @ 0.6µsec per scan,

transfer line 310°C

1. Triethylthiophosphodiate2. Thionazine3. Sufulfotep4. Phorate5. Dimetholate6. Diazinon7. Terbuphos8. Disulfoton9. Methyl parathion10. Malithion11.Parathion12. Merphos13.Fenamiphos14.Carboxin15.Famophos

7.5 10.0 12.5 15.00 17.5 minutes

0.0

0.5

1.0

1.5

2.0

2.5

MCounts

1

2

3

4

67

8

5

910

11

12

13

14 15

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Applications with SPME PEG FiberTriazine Herbicides at 2ppb by SPME

1 Desisopropyl atrazine

7 Simetryn

2 Desethyl atrazine

8 Ametryn

3 Prometon

9 Prometryn

4 Simazine

10 Terbutryn

5 Atrazine

11 Cyanazine

6 Propazine

Triazine pesticides at 2ppb in 0.1M Phosphate buffer, pH 7, with25% NaCl, 9mL in 10mL vial

Sample:

Extraction: 30min, direct immersion with agitation using CombiPal, Fibers: 60µm PEGDesorption: 2min at 250°C, splitless for 0.75min than opened

Column: SLB-5 MS 30m x 0.25mm ID, 0.5µm film Oven Program: 60°C(1min) to 150°C @25°C/min to 230°C @ 5°C/min (5min hold)Carrier gas: Helium, 1mL/min constant flow, Injector: Splitless/split with 0.75mmID liner and

Merlin MicrosealDetector ITMS m/z = 150-250 @ 0.5µsec per scan,

transfer line 310°C

1 2

3

4

5

6

7

8

9 10

11

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Comparison of Oxygenates in Gasoline Direct Injection vs. SPME (Chromatograms normalized)

0 0.5 1 1.5 2 2.5 3 3.5 4 4.5

min0 0.5 1 1.5 2 2.5 3 3.5 4 4.5

uV

0

5000

10000

15000

20000

25000

Ethanol

Butane

IPA

Pentane

n-Propanol

min

uV

0

50000

100000

150000

200000

Ethanol

Butane

IPA

Pentane

n-Propanol

SPME

Direct injection

15min direct immersion,5min desorption (240°C)

Split 50:1

Oxygenates in gasoline at 400ppm (0.4%)

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Enhancement of Oxygenates in Gasoline using SPME

Relative to Butane SPMESPME Injection SPME Injection Enhancement

Ethanol 21.214 2.055 0,493 0,003 155Isopropanol 12.403 2.191 0,288 0,003 85n-Propanol 16.212 2.636 0,376 0,004 92Butane IS 43.071 644.669

Area Counts

Oxygenates in gasoline at 400ppm (0.4%)

Direct injection of gasoline, 0.2µl, split 50:1

SPME Extraction 15min, direct immersion with PEG Fiber, Split 50:1

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Findings Concerning 60µm PEG Fiber

• Methanol may be produced by the fiber under acidic conditions for the first several extractions. (trace amount of inhibitor)

• The fiber can swell when exposed to water samples with total soluble organic content exceeding 200ppm. ->Best suited for trace level analyses.

• The fiber does not swell in hydrocarbon solvents - suitable for extraction of polar analytes out of hydrocarbon

solvents.

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Conclusions

• New more durable and flexible metal alloy assembly has been designed

• Super-elastic properties make it more compatible with autosamplers• Coatings on a new metal fiber core made with the same super-elastic

properties have been developed• The fiber core is inert, durable and the coating bonds well to it• New PEG fiber coating has been developed without an adsorbent

- The coat is durable and long lasting- PEG fiber is available in standard stainless steel assemblies- PEG fiber is suitable for extraction of polar analytes out of hydrocarbon

solutions

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Official Methods / Applications applying SPME

DIN 38407-F 34 (NEW)- Determination of selected plant treatment agents, biocides

and break-down products; Method using gas chromatography (GC-MS) after solid-phase micro extraction (SPME)

Other norms• ASTM D 6438, 2005 • ASTM D 6520, 2000 • ASTM D 6889, 2003 • ASTM E 2154, 2001 • OENORM A 1117, 2004-05-01

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DIN 38407-34 – Compounds Analyzed

1. Aldrin 12. Lindan 2. Atrazin 13. Mefenpyr-diethyl 3. Carfentrazon-ethyl 14. Metolachlor 4. Desethylatrazin 15. Metribuzin 5. Desethylterbutylazin 16. Parathion-ethyl 6. Dichlobenil 17. Parathion-methyl 7. Dieldrin 18. Pendimethalin 8. Diflufenican 19. Simazin 9. endo-Heptachlorepoxid 20. Terbutryn

10. exo-Heptachlorepoxid 21. Terbutylazin 11. Heptachlor 22. Triclosan

Method might be suitabel for others – need to be evaluated individially

Operational range of method the above 0.05µg/Ldepending on matrix

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DIN 38407-34 conditions

SPME Conditions- Fiber: Polyacrylate- Conditioning: Fiber 20 min @300°C- Extraction: pH 6-8, NaCl close to saturation (e.g. 2.4g/8mL sample)

60 min @ 30°C, stirring 250 rpm- Desorption: 10 min @ 280°,splitless (6 min)

GC conditions (example)- Injection KAS/CIS 60°C; 10°C/s to 280°C;10min;Splitless- Column 5% Phenyl, 30m x 0.25mm, 0.25µm- Carrier gas Helium (5.0), 0,9mL/min- Oven 60°C, 2 min; 4°C/min to 220°C; 10°C/min to 300°C- Detector MS, EI 70eV, SIM

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Other official methods for SPME

• ASTM D 6438, 2005- Standard Test Method for Acetone, Methyl Acetate, and

Parachlorobenzotrifluoride Content of Paints, and Coatings by Solid Phase Microextraction-Gas Chromatography

• ASTM D 6520, 2000- Standard Practice for the Solid Phase Micro Extraction (SPME) of Water

and its Headspace for the Analysis of Volatile and Semi-Volatile OrganicCompounds

• ASTM D 6889, 2003 - Standard Practice for Fast Screening for Volatile Organic Compounds in

Water Using Solid Phase Microextraction (SPME)• ASTM E 2154, 2001

- Standard Practice for Separation and Concentration of Ignitable Liquid Residues from Fire Debris Samples by Passive Headspace Concentration with Solid Phase Microextraction (SPME)

• OENORM A 1117, 2004-05-01 - Determination of volatile compounds in cellulose-based materials by Solid

Phase Micro Extraction (SPME)

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New SPME CD

• NEW! Video demo

• 600 new application references

• SPME Applications by Area - Environmental 720 - Food, Flavors and Fragrances 550 - Forensics/ Toxicology 243 - Pharmaceutical/Biologicals 150

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Thank You!

Seminar Offer

15% off on SPME Starter Kit

• 1 SPME Holder• 1 Box of SPME Fibers of choice• 1 SPME GC Injector Liner