Analysis of VOCs in complex samples

John R. Dean Northumbria University Email: John.Dean@northumbria.ac.uk

Ethylbutanoate
O O

Isovaleric acid (3-methylbutanoic acid) 1-Octen-3-one

O OH
O

Methional

VOCs
3-Methylindole (skatole)

2-Methoxyphenol (Guaicaol)
O

2-Phenylethanol OH

HO

N H

Ethylbutanoate
O O

Isovaleric acid (3-methylbutanoic acid) 1-Octen-3-one

O OH
O

Fruity, apple

Mushroom, metallic Methional

Sweaty

S
Boiled potato

VOCs
(60)

2-Methoxyphenol (Guaicaol)
O

2-Phenylethanol OH

3-Methylindole (skatole)

HO

Spicy, burnt, sweet

N H

Wine Roses, sweet

Faecal

Malodour on Laundry

Purge gas To GC Trap

Does malodour in washing machines cause/contribute to malodour on fabrics?

What are the key malodour compounds? On laundry In the washing machine

Overview of experimental
Direct olfactive analysis Microbiological tests Swabs of surfaces within machines Consumer washing machines DHS-GC-O/MS Fabrics washed in machines

Direct olfactive analysis Microbiological tests

HS-SPME-GC-O/MS

Microbiological tests

HS-SPME-GC-O/MS

Overview of experimental
Direct olfactive analysis Microbiological tests Swabs of surfaces within machines Consumer washing machines DHS-GC-O/MS Fabrics washed in machines

Direct olfactive analysis Microbiological tests

HS-SPME-GC-O/MS

Microbiological tests

HS-SPME-GC-O/MS

Machine

Definition
Brand new washing machine Used Washing Machine Consumer washing machine (Consumer 1) Consumer washing machine (Consumer 2)

Malodour?
No No Yes (Grade 8)

Make and Model

Hotpoint Aquarius WF541 1400 Miele Honeycomb Care W3622 Bosch Exxcel 1400 express, maxx freedom performance Hoover Optima OPH714D 7kg 1400

Cycle chosen
Cold wash, 800rpm, Woollen, 52 mins Cold wash, 800rpm, Woollen, 39 mins Cold wash, 800rpm, Woollen (with wash plus), 50 mins

1 2 3

Yes (Grade 6)

Cold wash, 800rpm, Woollen, 59 mins

Machine

Definition
Brand new washing machine Used Washing Machine Consumer washing machine (Consumer 1) Consumer washing machine (Consumer 2)

Malodour?
No No Yes (Grade 8)

Make and Model

Hotpoint Aquarius WF541 1400 Miele Honeycomb Care W3622 Bosch Exxcel 1400 express, maxx freedom performance Hoover Optima OPH714D 7kg 1400

Cycle chosen
Cold wash, 800rpm, Woollen, 52 mins Cold wash, 800rpm, Woollen, 39 mins Cold wash, 800rpm, Woollen (with wash plus), 50 mins

1 2 3

Yes (Grade 6)

Cold wash, 800rpm, Woollen, 59 mins

Olfactive Grade
Drawer

Machine 1
0

Machine 2
0

Machine 3
4

Machine 4
2

Rubber seal/Drum Sump

Coin Trap

0 0
0

1 2
0

8 8
4

6 6
3

No. of colony forming units (k) Drawer Rubber seal Sump

Machine 1
0 51 440

Machine 2
0 43 6.2

Machine 3
39.9

Machine 4
<0.05

620
750

256
475

Olfactive Grade
Drawer

Machine 1
0

Machine 2
0

Machine 3
4

Machine 4
2

Rubber seal/Drum Sump

Coin Trap

0 0
0

1 2
0

8 8
4

6 6
3

No. of colony forming units (k) Drawer Rubber seal Sump

Machine 1
0 51 440

Machine 2
0 43 6.2

Machine 3
39.9

Machine 4
<0.05

620
750

256
475

Olfactive Grade
Drawer

Machine 1
0

Machine 2
0

Machine 3
4

Machine 4
2

Rubber seal/Drum Sump

Coin Trap

0 0
0

1 2
0

8 8
4

6 6
3

Overview of experimental
Direct olfactive analysis Microbiological tests Swabs of surfaces within machines Consumer washing machines DHS-GC-O/MS Fabrics washed in machines

Direct olfactive analysis Microbiological tests

HS-SPME-GC-O/MS

Microbiological tests

HS-SPME-GC-O/MS

Olfactory grading of fabrics / Washing machine 3 (n = 3)

10

Olfactory grading (arbitrary scale)

9 8 7 6 5 4 3 2 1 0 0 10 20 30 40 50 60

Terry-towel Fleece Washing machine

Time (hours)

DHS-GC-O/MS of rubber seal of washing machine 3

Examples of peak identification*

peak number 11 GC-O Floral, fragrant GC-MS -pinene

14
22 28

cheese, sulphur
burnt, fatty, long-lasting sweet, floral, slightly burnt

dimethyl trisulphide
nonanal decanal

* Known perfume components (e.g. Eucalyptol) and surfactants (at long retention times) were also detected

Overview of experimental
Direct olfactive analysis Microbiological tests Swabs of surfaces within machines Consumer washing machines DHS-GC-O/MS Fabrics washed in machines

Direct olfactive analysis Microbiological tests

HS-SPME-GC-O/MS

Microbiological tests

HS-SPME-GC-O/MS

Microbiological identification of bacteria from washing machines

Analytical Profile Index (API) strips

and

MALDI-TOF-MS

Fabrics versus washing machines

62 different microbes identified from 4 washing machines 45 different microbes identified from washed fabrics
Majority of microbes were from environmental sources i.e. not pathogenic bacteria.

HS-SPME-GC-MS identification of VOCs from bacteria from washing machines and fabrics
10 VOCs commonly identified from bacteria from washing machines and fabrics (fleece and Terrytowel).
Dimethyl disulphide; 3-methyl-1-butanol; 2,4dithiapentane; dimethyl trisulphide; 2-tridecanone; indole; 2-phenylethanol; isovaleric acid; isobutryric acid; and, 1-undecene.

HS-SPME-GC-MS of Shewanella putrefaciens from washing machine

Peak identifications: (1) dimethyl disulphide; (2) siloxane; (3) 2,4-dithiapentane; (4) siloxane; (5) dimethyl trisulphide; (6) unknown; (7) 2-nonanone; (8) 2-undecanone; (9) isovaleric acid

NOTE: Shewanella putrefaciens is a Gram-negative bacteria found in marine environments; it has a rotting fish smell.

Conclusions
Evidence obtained that points to a possible link that contamination of fabrics occurs from washing machines.
Based on the identification of the same bacteria in WM rubber seal and on fabrics. Potential VOC markers for both high levels of bacteria and malodour in washing machines are dimethyl disulphide and dimethyl trisulphide.

Gas Chromatography-Ion Mobility Spectrometer

Multi capillary column

The MCC comprises a stainless steel tube, 20 cm x 3 mm ID, containing approximately 1200 parallel capillary tubes, 40 m ID, coated with 0.2 m film thickness of stationary phase.

Stationary phases: Carbowax 20M (polyethylene glycol) and OV-5 (5% - diphenyl, 95% - dimethylpolysiloxane)

http://www.mcc-chrom.com/

Working principle of the IMS

IMS Drift Tube Ionisation region Sample inlet + + + + + + + + + Electrical field Faraday plate Drift region

Driftgas inlet
BN-Shutter

Outlet

Working principle of the IMS

IMS Drift Tube Ionisation region Sample inlet Electrical field + ++ + + + + + + + + + Faraday plate Drift region

Driftgas inlet

Outlet

BN-Shutter

Working principle of the IMS

IMS Drift Tube Ionisation region Sample inlet Electrical field Drift region

+
+ + Outlet BN-Shutter +

+ + + +

+ Faraday plate + + + + Driftgas inlet

Signal

Drift time / ms

Drift time (ms) Retention time (s)

Formation of Reactant Ion Peak (RIP):

N2
N2+ + (H2O)m H+(H2O)m-1

N2+ + eH+(H2O)m-1 + OH- + N2 H3O+(H2O)m-2 RIP

Formation of monomer, dimer and trimer: X + H3O+(H2O)m-2 XH+(H2O)m-3 + H2O

[XH+(H2O)m-3
X + XH3O+(H2O)m-4

XH3O+(H2O)m-4]
X2H+(H2O)m-5 + H2O

[X2H+(H2O)m-5
X + X2H3O+(H2O)m-6

X2H3O+(H2O)m-6]
X3H+(H2O)m-7 + H2O

[X3H+(H2O)m-7 etc]

Drift time (ms) Retention time (s)

2-Dimensional map of 14 VOCs

400

4-methylvaleric acid

350

Retention time (s)

300

250

dimethyl trisulfide

200

150

100

50

0 6 7 8 9 10 11 12 13 14 15

Drift time (ms)

Typical operating conditions

Compounds in mixture: ethylbutanoate, octanal, E-2-octanal, E-2-nonanal, (E,E)-2,4-nonadienal, (E,E)-2,4-decadienal, 1-decanol, 3-methylindole, 4methoxybenzaldehyde, guaiacol, 2-phenylethanol, 1-dodecanol Column: Carbowax (polar) Carrier gas flow rate: 150 mL/min Drift gas flow rate: 500 mL/min Column temperature: 30 - 80 oC IMS temperature: 45 oC RIP voltage: Positive Trigger duration: 100 s Electric field along IMS tube: 2 kV Incubation time: 10 min Incubation temperature: 95 oC Sample in Vial: 2 mL Injection volume: 2.2 mL Measurement run time: 11 20 mins Syringe temperature: 95 oC

Conclusions: GC-IMS
Potential for rapid screening of VOCs in samples. Potential to detect small S-, N-containing VOCs.
Further work required on authentic samples

Acknowledgements