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. 2023 Jan 20;10:195–205. doi: 10.1016/j.toxrep.2023.01.008

Table 2.

Studies reporting original data from preliminary toxicological assessment of heated tobacco products (HTP) according to the types of studies performed, categories of HTPs, and comparator reference products.

Author-date Aerosol Chemistry In Vitro In Vivo Other HTP
typea
Reference cigaretteb Test material generatione Analytes / Assaysp
Zenzen 2012 X X E 2R4F
CC
ISO
HPBf
2) 49 HPHCs
3) Ames / NRU
Schaller 2016a X X X E 3R4F HCI
ISO
APRg
2) 58 HPHCs
3) Ames / NRU / MLA
5) aerosol droplet size - MMAD
Oviedo 2016 X E 3R4F HCI 4) OECD Test No. 413 /Systems toxicology endpoints
Schaller 2016b X E 3R4F HCI 2) 58 HPHCs
Wong 2016 X E 3R4F HCI 4) OECD Test No. 413 /Systems toxicology endpoints
Auer 2017 X E CC;
PDc
ISO 2) 8 VOCs; 16 PAHs; 3 inorganic compounds; Nicotine
Bekki 2017 X E 1R5F;
3R4F
HCI 2) Nicotine; Tar; CO; 4 TSNAs
Farsalinos 2017 X E CC HCI
LPVh
2) Nicotine
Jaccard 2017 X E 3R4F;
CC
HCI 2) 44 compounds (Health Canada list)
Poynton 2017 X A 3R4F HCI
CRM81i
2) Untargeted GC scans;
113 compounds (FDA / HC)
Jaunky 2018 X E 3R4F HCI 3) NRU
Takahashie 2018 X X A 3R4F HCI 2) 43 Hoffmann analytes; propylene glycol, glycerol, triacetin, TPM, Nicotine, CO;
3) Ames / MN / NRU
Thorne 2018 X E 3R4F HCI 3) Ames / NRU / MLA / Bhas cell transformation
Eaton 2018 X E 3R4F nak 1) TGAq
Farsalinos 2018 X E CC HCI
+ 2 Intensel
2) 5 carbonyls (acetaldehyde, acrolein, formaldehyde, propionaldehyde and crotonaldehyde)
Forster 2018 X X E 3R4F HCI
CRM81i
2) 126 HPHCs
5) Physical analysis: particle diameter and number
Li 2018 X X E 3R4F HCI
ISO
1) Simulated pyrolysis
2) TPM; water, tar, nicotine, propylene glycol, glycerin, CO, VOCs, aromatic amines, HCN, ammonia, TSNAs, phenol, PAHs
Mallock 2018 X E PDd HCI 2) TPM; Nicotine, Water, Aldehydes (Acetaldehyde; acrolein; formaldehyde; crotonaldehyde) VOCs (1,3-Butadiene; Benzene; Isoprene; Styrene; Toluene)
Pacitto 2018 X E PD (-/2/10)m 5) physical characteristics of the aerosol (Particle size distribution; particle volatilitiy)
Savareear 2018 X E 3R4F HCI 2) Non targeted screening of VOCs from PP fraction of aerosols
Titz 2018 X C 3R4F HCI 4) OECD413 / Systems toxicology endpoints
Godec, 2019 X E 3R4F HCI 3) Ames / MLA
Thorne 2019a X E 3R4F HCI 3) MLA
Thorne 2019b X E 3R4F HCI 3) MN
McAdam 2019 X C, E CC Multiplen 2) Total aerosol mass (AM); smoke particulate matter (TPM); Water; Nicotine; Glycerol
Salman 2019 X X E CC HCI
ISO
2) Carbonyl compounds and total nicotine (free-base and protonated) and Reactive oxygen species (ROS)
Savareear 2019 X E 3R4F HCI
ISO°
2) non-targeted screening for PP components
Caruso 2021 X E IR6F HCI 3) Cytotoxicity tests in Human H292 cells via ALI (Air-liquid interface) exposure:
NRU; MTT; Annexin V apoptosis;
Dusautoir 2021 X X E 3R4F HCI (WA) 2) Nicotine; Aldehydes, PAHs
3) BEAS-2B cells cell viability; Glutathione Content assay; Gene expression; inflammatory mediators (GM-CSF; GRO-α IL-1ẞ; IL-6; IL-8; IL-13; MCP-1; MIP-1 α, RANTES and INF-γ)

a: HTP category as defined by CORESTA [23]: E = electrically heated tobacco product (eHTP); A = aerosol-heated tobacco product (aHTP); C = carbon-heated tobacco product (cHTP).

b: CC = marketed conventional cigarettes, PD = published data.

c: Published data based on reference [71].

d: Published data based on reference [12]

e: Test material generation as defined by the puffing regimen applied to produce the testing material characterized by puff volume (mL)/puff duration(s)/puff interval(s): ISO = International Organization for Standardization (35/2/60); HCI = Health Canada Intense (55/2.0/30). Other regimens specified by relevant footnotes: CRM81 = CORESTA recommended method (55/3/30); test material exposure; TPM = total particulate matter; GVP = gas–vapor phase; WA = whole aerosol; ALI = air–liquid interface.

f: HPB = human puffing behavior—nine different puffing regimens compared based on observed human topography studies (see [76] for details).

g: APR—test material also generated using five alternative puffing regimens based on human puffing behavior (see [56] for details).

h: LPV includes a long puff variant of the HCI regimen (55/4/30) in addition to the standard HCI regimen.

i: Reference cigarette 3R4F generation using HCI regimen; CRM81 for HTPs.

k: na = not applicable. A pyrolysis study was conducted based on 10 g of tobacco with 10 g wrapping paper heated in air or nitrogen in a platinum crucible up to 900 °C (see [16] for experimental details).

l: Products were tested using HCI and two more intense regimens (80/3/30 and 90/3/25).

m: Each test was performed with four puff profiles of five puffs for 2 s and an inter-puff time of 10 s. The puff volume was not specified; however, the flow rate was 1 L/min.

n: Five different regimens used with fixed 30-s puff interval and 20 puffs per device. The puff volume ranged from 20 mL to 150 mL and the duration from 2 s to 5 s

o: HCI regimen used for the HTPs; ISO for 3R4F.

p:1) Brief description of the pyrolysis method applied; 2) analytes measured in aerosol chemistry studies represented as the total number of analytes; plus, details of specific lists as given by the authors; 3) in vitro assays used: NRU = neutral red uptake, MLA = mammalian mouse lymphoma assay, MN = micronucleus, other tests as described; 4) in vivo protocol applied: OECD413 = Organization for Economic Cooperation and Development Test Guideline 413: Repeated dose inhalation toxicity; 5) other assays are briefly described. Please refer to the original papers for full details.

q: TGA = thermogravimetric analysis with Pyris 1 TGA system in air or nitrogen: 5 °C/min from ambient temperature to 240 °C, held for 5 min at 240 °C, and continued ramping to 900 °C.