according to the respective OECD guideline 471 (OECD, 1997a) as
previously described (Roemer et al., 2012).
Determinations were performed in the presence and in the
absence of a metabolic activation system.
2.7. In vitro mammalian cell genotoxicity
Genotoxicity of TPM was assessed in the Mouse Lymphoma TK
Assay (MLA) developed by Clive et al. (1972). It was performed
according to OECD guideline 476 (OECD, 1997b) in the microtiter
plate version (Cole et al., 1986) with L5178Y cells with and without
metabolic activation as described in detail by Schramke et al.
(2006).
2.8. Statistics
Results obtained for the test cigarette groups were compared to
the control cigarette group with standard statistical tools. The reference cigarette 2R4F was only used as an internal reference to
monitor consistency with historical data of the laboratory and
was not included in the statistical evaluation.
3. Results
3.1. Smoke chemistry
The following smoke constituents were below the limit of quantitation (LOQ) in smoke from control and/or test kretek cigarettes,
and consequently they will not be discussed further: N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), Nnitrosodiethylamine (NDEA), N-nitrosodi-n-propylamine(NPRA),
according to the respective OECD guideline 471 (OECD, 1997a) aspreviously described (Roemer et al., 2012).Determinations were performed in the presence and in theabsence of a metabolic activation system.2.7. In vitro mammalian cell genotoxicityGenotoxicity of TPM was assessed in the Mouse Lymphoma TKAssay (MLA) developed by Clive et al. (1972). It was performedaccording to OECD guideline 476 (OECD, 1997b) in the microtiterplate version (Cole et al., 1986) with L5178Y cells with and withoutmetabolic activation as described in detail by Schramke et al.(2006).2.8. StatisticsResults obtained for the test cigarette groups were compared tothe control cigarette group with standard statistical tools. The reference cigarette 2R4F was only used as an internal reference tomonitor consistency with historical data of the laboratory andwas not included in the statistical evaluation.3. Results3.1. Smoke chemistryThe following smoke constituents were below the limit of quantitation (LOQ) in smoke from control and/or test kretek cigarettes,and consequently they will not be discussed further: N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), Nnitrosodiethylamine (NDEA), N-nitrosodi-n-propylamine(NPRA),
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