structure-activity relationships of propylene glycol, glycerin ......3-hydroxypropanal background...

Altria Client Services l Matt Melvin l October 23, 2018 l CORESTA 2018 ST05 l 1

Melvin, M.S.; Ballentine, R.M.; Gardner, W.P.; McKinney, W.J.; Smith, D.C.; Pithawalla, Y.B.; Wagner, K.A.

Structure-Activity Relationships of Propylene Glycol, Glycerin, and Select Analogs for Carbonyl Thermal Degradation Products

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Thermal Degradation of eLiquids


HeatPropylene Glycol

Glycerin

Nicotine

Flavor Systems

▪ Propylene glycol (PG) and Glycerine (GLY) can thermally

degrade upon heating

- Formaldehyde, Acetaldehyde, Acrolein1,2,3

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Carbonyls in E-Cigarettes

▪ Geiss et al. and Gillman et al. demonstrated that carbonyl

formation increased with temperature1,4

▪ US FDA PMTA Draft Guidance for ENDS Products recommends

reporting four carbonyls in e-liquid and aerosol5


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Objectives and Approach

▪ Determine the formation pathways of formaldehyde,

acetaldehyde, acrolein, and crotonaldehyde:

1. Identify source of degradation products using 13C3-labeled PG

and GLY

2. Determine the role of 3-hydroxypropanal (3-HPA) as an

intermediate during the thermal degradation of e-liquids

3. Propose rational mechanisms based on results

4. Determine key reaction centers using rationally selected

derivatives of PG and GLY


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Microwave Model System

▪ Model microwave system used to generate

target carbonyls- Previously used to identify diacetyl and acetyl

propionyl formation pathways6

▪ Microwave system evaluated for equivalent

yields to e-cigarette- Sample = 50% PG : 50% GLY + 2.5 % nicotine (w/w)

- 140 puffs

- 55 mL puff volume, 5 sec puff duration, 30 sec puff

period, square wave


CEM Discovery SP Hybrid

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Analyte Yield Comparison


140 puffs using 55 ml Puff Volume, 5 sec Puff Duration, 30 sec Puff Period; Square Wave

0

5

10

15

20

25

30

Formaldehyde Acetaldehyde Acrolien

An

aly

te A

mo

un

t (µ

g /

g)

Liquid

Microwave

Aerosol

Crotonaldehyde was not detected

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A


Identify Source of Degradation Products

Using 13C-labeled PG and GLY

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Carbon-13 Labeled PG and GLY

▪ Samples:

- 50% 13C3-PG : 50% GLY + 2.5% nicotine (w/w)

- 50% PG : 50% 13C3-GLY + 2.5% nicotine (w/w)


MicrowaveHeating*

DNPH Derivatization

UPLC-UV-MS/MS

Isotope Distribution

*500 mg of sample heated to 180 °C and held for 3 min

▪ Labeled products directly traceable to labeled precursor

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Product Distribution Using 13C3-PG


50% 13C3-PG : 50% GLY + 2.5 % Nicotine (w/w)

26.1

4.7

0.7

0

5

10

15

20

25

30C

on

ce

ntr

ati

on

(µ

g / g

)

25.1%

93.2% 94.0%

74.9%

6.8% 6.0%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

Pe

rce

nta

ge

of

To

tal A

na

lyte

13C-PG

GLY

13C 13C13C


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Product Distribution using 13C3-GLY


50% PG : 50% 13C3-GLY + 2.5 % Nicotine (w/w)

22.7%

87.6%

95.3%

77.3%

12.4%

4.7%

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

Formaldehyde Acetaldehyde Acrolein

Pe

rce

nta

ge

of

To

tal

An

aly

te

PG

13C-GLY


13C 13C

13C

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Summary: 13C-Labeling Studies

▪ Formaldehyde was predominantly formed from GLY

▪ Acetaldehyde and acrolein were predominantly formed

from PG

▪ Crotonaldehyde was not detected


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Determine the Role of 3-hydroxypropanal

(3-HPA) as an Intermediate During the

Thermal Degradation of e-Liquids


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

3-Hydroxypropanal Background

▪ Researchers proposed formaldehyde and acetaldehyde are

produced from the retro-aldol condensation of

3-hydroxypropanal (3-HPA)4,7


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

3-HPA Fortification Studies


500 mg e-liquid 50% PG : 50% GLY + 2.5 % nicotine (w/w)

Fortify samples with 3-HPA at 3 levels (300, 700, 1500 µg)

Microwave Heating: 180 °C for 3 min

DNPH Derivatization

UPLC-UV-MS/MS Analysis

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Results: 3-HPA Fortification (N=3)


50% PG : 50% GLY + 2.5 % Nicotine (w/w)

0

100

200

300

400

500

600

Unfortified 350 700 1400

An

aly

te A

mo

un

t (µ

g /

g)

3-HPA Fortification (µg)

Formaldehyde Acetaldehyde Acrolein

Acrolein Yield ~ 30%

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Summary: 3-Hydroxypropanal (3-HPA)

▪ Unfortified e-liquids- 3-HPA, acrolein, and crotonaldehyde were not detected


▪ E-liquids fortified with 3-HPA- Crotonaldehyde was not detected

- No increase in formaldehyde and acetaldehyde

- 3-HPA converted to acrolein with ~30 % yield

➢The retro-aldol condensation of 3-HPA appears to be a negligible

pathway for the production of formaldehyde and acetaldehyde under

test conditions

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Suggested Formation Pathways in Aerosol3-HPA was not detected


▪ Formaldehyde from Glycerin

▪ Acetaldehyde from Propylene Glycol

▪ Acrolein from Propylene Glycol

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Determine Key Reaction Centers

Using Rationally Selected

Derivatives of PG and GLY


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Experimental: Evaluation of Derivatives

▪ Derivatives:

- Methoxy derivatives selected to reduce autoxidation efficiency

- Methyl derivatives selected to reduce dehydration efficiency

▪ Samples:

- 50% PG : 50% GLY-Deriv + 2.5 % nicotine (w/w) -> Formaldehyde

- 50% PG-Deriv : 50% GLY + 2.5 % nicotine (w/w) -> Acetaldehyde and

Acrolein

▪ Control = 50% PG : 50% GLY + 2.5% nicotine (w/w)


Microwave Heating:

180 °C for 3 min

DNPH Derivatization

UPLC-UV-MS/MS

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

GLY Derivatives: Formaldehyde


Methoxy Derivatives

Methyl Derivatives

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Formaldehyde: GLY Derivatives


Results support proposed mechanism

0

10

20

30

40

50

Control D E G F H N O P R Q

Fo

rmald

eh

yd

e (

µg

/ g

)

Control

Methoxy Derivatives

Methyl Derivatives

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

PG Derivatives: Acetaldehyde and Acrolein


Methoxy Derivatives

Methyl Derivatives

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Acetaldehyde: PG Derivatives Results do not support proposed mechanism


0

10

20

30

40

Control A B C I M J K L

Ac

eta

lde

hyd

e (

µg

/ g

)

Control

Methoxy Derivatives

Methyl Derivatives

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Acrolein: PG DerivativesResults support proposed mechanism


0.00

0.10

0.20

0.30

0.40

Control A B C I M J K L

Ac

role

in (

µg

/ g

)

Control

Methoxy Derivatives

Methyl Derivatives

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Summary: Methoxy and Methyl Derivatives▪ Formaldehyde: GLY Derivatives

- Substitution reduced formaldehyde generation

- Consistent with proposed pathway

▪ Acetaldehyde: PG Derivatives

- Substitution increased acetaldehyde production

- Not consistent with proposed pathway

- Under further investigation

▪ Acrolein: PG Derivatives

- Substitution decreased acrolein generation

- Consistent with proposed mechanism

▪ Crotonaldehyde was not detected


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

Conclusions

▪ Formaldehyde derived primarily from glycerin

▪ Acetaldehyde and acrolein derived primarily from propylene glycol

▪ 3-hydroxypropanal pathway has negligible contribution to

formaldehyde and acetaldehyde generation

▪ Proposed pathways for formaldehyde and acrolein are consistent with

experimental results

▪ Acetaldehyde pathway under further investigation


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A

References:1. Geiss, O., Bianchi, I., and Barrero-Moreno, J. (2016) Correlation of volatile carbonyl yields emitted by e-cigarettes with the temperature of the

heating coil and the perceived sensorial quality of the generated vapours. Int J Hyg Environ Health 219, 268-277.

2. Farsalinos, K. E., and Gillman, G. (2017) Carbonyl Emissions in E-cigarette Aerosol: A Systematic Review and Methodological

Considerations. Front Physiol 8, 1119.

3. Kosmider, L., Sobczak, A., Fik, M., Knysak, J., Zaciera, M., Kurek, J., and Goniewicz, M. L. (2014) Carbonyl compounds in electronic

cigarette vapors: effects of nicotine solvent and battery output voltage. Nicotine Tob Res 16, 1319-1326.

4. Gillman, I. G., Kistler, K. A., Stewart, E. W., and Paolantonio, A. R. (2016) Effect of variable power levels on the yield of total aerosol mass

and formation of aldehydes in e-cigarette aerosols. Regul Toxicol Pharmacol 75, 58-65.

5. FDA. (2016). Guidance for industry. Premarket Tobacco Product Applications for Electronic Nicotine Delivery Systems. Draft Guidance.

Available at: https://www.fda.gov/downloads/TobaccoProducts/Labeling/RulesRegulationsGuidance/UCM499352.pdf. Accessed 15Feb2018.

6. Melvin, M.S.; Avery, K.C.; Ballentine, R.M.; Gardner, W.P.; McKinney, W.J.; Smith, D.C.; Wagner, K.A. Thermal Degradation Studies of

Electronic Cigarette Liquids Part 2: Development of a Model Reaction System Used to Study α-Dicarbonyl Formation. Presented at the 71st

Tobacco Science research Conference, 2017, Bonita Spring, Fl.

7. Flora, J. W., Meruva, N., Huang, C. B., Wilkinson, C. T., Ballentine, R., Smith, D. C., Werley, M. S., and McKinney, W. J. (2016)

Characterization of potential impurities and degradation products in electronic cigarette formulations and aerosols. Regul Toxicol Pharmacol

74, 1-11.


2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A


For copies of this presentation visit the Altria’s Science

Website at www.altria.com/alcs-science

2018

_ST

05_M

elvi

n.pd

fC

ongr

ess2

018

- D

ocum

ent n

ot p

eer-

revi

ewed

by

CO

RE

ST

A
http://www.altria.com/alcs-science/pages/gate-page.aspx?r=/alcs-science/Pages/default.aspx

structure-activity relationships of propylene glycol, glycerin ......3-hydroxypropanal background...

Documents