Dustin Tate Yeatman, MS Nicholas B. Tiscione, MS Ilene Alford, MS Dustin Tate Yeatman, MS Xiaoqin...

QUANTITATION OF ETHANOL AND

IDENTIFICATION OF OTHER VOLATILES BY HS-

GC-FID/MS

Dustin Tate Yeatman, MS

Nicholas B. Tiscione, MS

Ilene Alford, MS

Dustin Tate Yeatman, MS

Xiaoqin Shan, PhD, Palm Beach County Sheriff's Office

Joe Kahl, BS, Miami-Dade Medical Examiner Department

Objective Replace HS-GC dual

column, dual FIDPE HS40XL with

Autosystem XL○ 40 vial capacity, 20mL

HS vialsColumn A: J&W DB-

ALC1 (30m x 0.53mm)Column B: J&W DB-

ALC2 (30m x 0.53mm)

Objective Add confirmation of

volatiles by MS Maintain robust

quantitation of ethanol by FID

Single method with a single instrument for both BAC and inhalants

Objective Single column GC

J&W DB-ALC1○ 30m x 0.32mm

Split to FID and MSFID for BAC QuantMS for confirmationCapillary flow

technology (CFT) splitter

J&W DB-ALC1 J&W DB-ALC2

1 1

22

3 3

44

55

1.Methanol2.Ethanol3.Isopropanol

4.Acetone5.n-Propanol

Instrumentation Agilent System

G1888 HS7890A GC

○ FID○ Aux EPC

5975C VL○ Diffusion pump○ Triple Axis Detector

CFT Two way splitter with makeup gas

NIST 2008 MS Library

Purchased ~ $113 k

Agilent 2mm ID Deactivated Splitless Liner (5181-8818)

Capillary Flow Technology (CFT) 2 way splitter with makeup gas FID Restrictor: 1.060m x 0.18mm, 2mL/min constant flow MS Restrictor: 2.890m x 0.18mm, 2mL/min constant flow

J&W DB-ALC1 column (30m x 0.32mm)

Method Details Tiscione, N.B.,

Yeatman, D.T., Shan, X., Kahl, J. (2013) Identification of Volatiles by Headspace GC with Simultaneous FID and MS Detection. Journal of Analytical Toxicology. 37, 573-579. *High doses

Method Highlights Desk fans used to

accelerate GC cooling

20 mL HS vials Maximum of 40 vials HS Oven

50°C for 20 min. 5:1 Split Injection Flow at 2mL/min GC Oven program

35°C for 2 minutes then 25°C per minute to 90°C with a final hold time of 4.3 minutes

8.5 min. GC run time 13 min. GC cycle time

EthanolValidation

Sample volume 100uL w/1mL IS

LOD 0.005 g/dL

LOQ 0.010 g/dL

Linear range 0.010 – 1.0 g/dL

CV 0.010 g/dL 0.025 g/dL 0.080 g/dL 0.300 g/dL

< 3.1 %< 0.7 %< 0.6 %< 0.5 %

Accuracy 0.010 g/dL 0.025 g/dL 0.080 g/dL 0.300 g/dL

± 8 %± 6 %± 3 %± 1.5 %

EthanolValidation and Method Application

Specificity/Selectivity 34 compounds

Case Comparison Ethanol > 0.02 Agreement

81 59 (73%) within ±0.0068 g/dL

Blood Cases Ethanol > 0.02

> 1076 754 (70%)

ATP Proficiencies 124 samples, 4 analysts

Typical replicates ± 0.004 g/dL

Uncertainty @99.8% kcorr = 3.16 n = 160 of 0.025 g/dL

± 9.76 %

J&W DB-ALC1Old Method

J&W DB-ALC1New Method

1

23

4

5

1.Methanol2.Ethanol3.Isopropanol

4.Acetone5.n-Propanol

1

2 3

45

123

4

5

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.000

20000

40000

60000

80000

100000

120000

140000

160000

180000

200000

220000

240000

260000

280000

300000

Time-->

Abundance

Signal: NBT33.D\FID1A.ch

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.0020000

40000

60000

80000

100000

120000

140000

160000

180000

200000

220000

240000

Time-->

Abundance

TIC: NBT33.D\data.ms

20 22 24 26 28 30 32 34 36 38 40 42 44 46 480

500

1000

1500

2000

2500

3000

3500

4000

m/z-->

Abundance

Scan 104 (1.943 min): NBT33.D\data.ms (-98) (-)31.0

29.0

16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 540

2000

4000

6000

8000

10000

12000

14000

16000

18000

m/z-->

Abundance


45.0

27.0

42.0

18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 680

2000

4000

6000

8000

10000

12000

14000

16000

18000

20000

22000

24000

26000

28000

30000

32000

34000

36000

38000

40000

42000

44000

m/z-->

Abundance


41.027.0

59.031.038.1

16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 680

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

110000

120000

130000

m/z-->

Abundance


58.0

27.1 39.036.0 53.031.024.1

16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 62 64 66 68 700

2000

4000

6000

8000

10000

12000

14000

16000

18000

20000

22000

24000

26000

28000

30000

32000

34000

36000

38000

40000

42000

44000

46000

48000

50000

m/z-->

Abundance


59.0

42.027.0

39.0

45.054.9

Methanol Ethanol

Isopropanol Acetone

n-propanol

InhalantsValidated for Qualitative Identification*

Methanol Acetone Acetaldehyde Toluene

MEK Isoamyl Alcohol Isobutyl Alcohol n-butyl alcohol

DFE TFE Norflurane Chloroethane

trichlorofluoromethane (Freon®-11)

dichlorodifluoromethane (Freon®-12)

dichlorofluoromethane (Freon®-21)

chlorodifluoromethane (Freon®-22)

1,2-dichlorotetrafluoroethane (Freon®-114)

*Validation included evaluation of matrix effects, sensitivity, carryover, specificity, repeatability, and ruggedness/robustness

DFE Case DUI Crash

Driving Pattern○ At stoplight, car immediately

behind honked horn to get driver to go when green.

○ Accelerated slowly at first then rapidly

○ Drove into median, over bushes and crossed other lanes

○ Drove through 2 yards and struck 2 parked cars

○ No attempt to brake or avoid collisions

Behavior○ Lethargic○ Used police car for balance

Driver Statements Traveling from Walmart

where she just purchased computer duster

Can of duster found in car Admitted to “huffing”

while stopped at red light Toxicology

Blood collected ~ 50 minutes after the accident

Identified: ○ 1,1-difluoroethane (DFE)○ THC < 2.5 ng/mL○ THCA at 4 ng/mL

DFE Case

20 25 30 35 40 45 50 55 60 65 70 750100020003000400050006000700080009000

10000110001200013000140001500016000

m/z-->

Abundance Scan 68 (1.655 min): DFE Blood Case\data.ms (-62) (-)

51.0

65.0

47.0

26.1 33.0

15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 1000100002000030000400005000060000700008000090000

100000110000

m/z-->

Abundance Scan 68 (1.655 min): DFE Control\data.ms (-61) (-)51.0

65.0

45.0

27.033.0 94.0

Case MS

Control MS

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00020000400006000080000

100000120000140000160000180000200000220000

Time-->

AbundanceSignal: DFE Blood Case\FID1A.ch

n-P

ropa

nol

1,1-

Difl

uoro

etha

ne

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.002000030000400005000060000700008000090000

100000110000120000130000140000150000160000

Time-->

AbundanceTIC: DFE Blood Case\data.ms

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.000100020003000400050006000700080009000

10000110001200013000140001500016000

Time-->

AbundanceIon 51.00 (50.00 to 52.00): XS03.D\data.ms

Ion 65.00 (64.00 to 66.00): XS03.D\data.ms

Ion 47.00 (46.00 to 48.00): XS03.D\data.ms

Case FID

Case TIC

Case EIC

Toluene Case DUI

Driving Pattern○ Suspect sitting in car

inhaling from bottle○ Stopped by police when

backing out of parking spot Behavior

○ Odor of contact cement○ Can in car○ Cement on nose, in

mouth, on lips, chin and neck

○ Extremely lethargic○ Slurred speech○ Unsteady on feet

SFSTs Did not follow

instructions Problems with balance

Toxicology Blood collected ~

1.3 hours after the stop

Identified: ○ Toluene○ Methyl Ethyl

Ketone (MEK)*

*MEK not confirmed—not validated at time of reporting

Toluene Case

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00020000400006000080000

100000120000140000160000180000200000220000240000

Time-->

Abundance

Signal: Toluene Blood Case\FID1A.ch

Tolu

ene

n-P

ropa

nol

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00100002000030000400005000060000700008000090000

100000110000120000

Time-->

AbundanceTIC: Toluene Blood Case\data.ms

5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 950100020003000400050006000700080009000

m/z-->

Abundance Scan 416 (4.519 min): Toluene Blood Casedata.ms43.0

72.128.0 57.050.0 88.0

5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 950100020003000400050006000700080009000

m/z-->

Abundance #648: 2-Butanone (Methyl Ethyl Ketone)

43.0

72.029.057.015.0 50.037.0

20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 10002000400060008000

10000120001400016000180002000022000240002600028000300003200034000360003800040000420004400046000480005000052000

m/z-->

Abundance Scan 652 (6.462 min): Toluene Blood Case\data.ms (-644) (-)

91.1

65.139.1 51.045.027.1 74.0 85.0

20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 1000100020003000400050006000700080009000

10000110001200013000140001500016000170001800019000200002100022000

m/z-->

AbundanceScan 652 (6.462 min): Toluene Control\data.ms (-645) (-)

91.1

65.039.1 51.045.027.1 74.1

Case FID

Case TIC

MEK MS--Case

MEK MS--Library

Toluene MS--Case

Toluene MS--Control

ME

K

Acknowledgements

Ilene Alford, Nick Tiscione, Xiaoqin Shan, and Joe Kahl

Palm Beach County Sheriff’s Office

[email protected]

mailto:[email protected]

Dustin Tate Yeatman, MS Nicholas B. Tiscione, MS Ilene Alford, MS Dustin Tate Yeatman, MS Xiaoqin...

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