Abstract
This is yet another scientific report generated for Philip Morris by INBIFO on the effects of secondhand smoke exposure. The date of this report is unclear, but it references another study that was done at INBIFO in 1982, which places it after that date. It also states the date the "test substances" (cigarettes) were received at INBIFO (15 October 1982), placing the report near and after that date. This report, like the others done on this topic, was forwarded to Dr. Ragnar Rylander, who worked on contract to the Philip Morris tobacco company.
In this experiment, one batch of rats were made to breathe various concentrations of secondhand smoke (given in a single "acute exposure" of 7 hours) and other rats were similarly exposed only to ammonia vapor. The objective was to "establish a dose-response relationship for the irritative capacity of sidestream cigarette smoke in comparison to pure ammonia vapor, which is supposed to be 1 of the main irritative factors in sidestream smoke."
The "Conclusions" section of the report states that diluted secondhand tobacco smoke is ten times more irritating than ammonia vapor alone:
"Inhalation of diluted sidestream smoke leads to strong short and long-term irritation...By and large, diluted sidestream smoke is 10 times more irritative than one would predict from its ammonia concentration."
The report concludes that "Ammonia is not the only irritative factor responsible for the relatively (as compared to mainstream smoke) high irritative capacity of sidestream smoke." It obligingly lists a few of the other chemicals in secondhand smoke which may be the cause of this high irritative capacity, specifically formaldehyde and acreolin which "seem to play an important role as far as the irritative capacity of sidestream smoke in concerned."
Page 21 of the report (Bates No. 2501592215) states the "test substances" used. The "cigarette code" is 2R1 and is a "standard reference" cigarette supplied by the Philip Morris. While no brand name is mentioned, the report does state that the 4,600 cigarettes used in the experiment arrived packed in "cartons with 200 cigarettes [of] 10 packages with 20 cigarettes/package," which is the same type of packaging used with their commercial brands.
The report indicates that by 1983 PM was in possession of detailed information describing the higher irritative characteristics of diluted secondhand smoke as compared to mainstream smoke.
Fields
- Notes
This document is 143 pages long, but the operative information is contained within the first 20 pages or so.
- Quotes
[From page 9, Bates No. 2501592203]:
In this acute inhalation study the influence of various concentrations of SIDESTREAM CIGARETTE SMOKE and of AMMONIA VAPOR on the body weight development, the food consumption, the respiratory frequency, the carbondioxide production and the rectal temperature of head-only-exposed rats and the correlation between these parameters was investigated... The aim of this l-day inhalation study was to establish a dose-response relationship for the irritative capacity of sidestream cigarette smoke in comparison to pure ammonia vapor, which is supposed to be 1 of the main irritative factors in sidestream smoke. Additionally, this study serves as a range finding study to future 21-day inhalation studies with the same test compounds. Accordingly, the applied sidestream doses covered the range between the no-effect level and a sidestream dose which was 2 times higher than the maximally tolerated daily sidestream total particulate matter (TPM) dose applicable in 21-day studies. Rats were kept in exposure tubes of a head-only exposure chamber. They were exposed for 7 hours to a constant concentration of sidestream smoke of standard reference cigarette 2R1 or to pure ammonia vapor.
[From Page 14, Bates No. 2501592208]:
1.4.3 Conclusions
From those end points used to access sensory irritation the determination of the respiratory frequency and the C02 exhalation are most sensitive. Both measure the ventilation and indirectly the metabolic rate and both record an immediate and a corresponding long-term effect with similar sensitivity. On the other hand, temperature, body weight development and food consumption only record long-term effects (i. e. metabolic rate) with similar sensitivity. Inhalation of diluted sidestream smoke leads to strong short and long-term irritation. Compared to that, inhalation with pure ammonia in concentrations similar to the ammonia concentrations in diluted sidestream smoke leads to relatively small irritation as far as short-term effects are concerned and to no apparent long-term effects. By and large, diluted sidestream smoke is 10 times more irritative than one would predict from its ammonia concentration. Ammonia therefore is not the only irritative factor responsible for the relatively (as compared to mainstream smoke) high irritative capacity of sidestream smoke.
...Other candidates for irritative gas/vapor phase components are listed in TABLE A...Although this table is not complete...formaldehyde and acreolin seem to play an important rold as far as the irritative capacity of sidestream smoke is concerned.
- Company
- Philip Morris
- Author
- Pruhs, D.
- Reininghaus, Wolf (INBIFO, Gen. Mgr., Contract Research)
Institut fur Biologische Forschung, Cologne, Germany - (Philip Morris' offshore research lab).
- Romer, Ewald (Quality Assurance Biologist, INBIFO, c. 1987)
- Schnell, P.
- Speck, M.
- Teredesai, A. (Pathologist, INBIFO, c. 1987)
- Recipient
- Rylander, Ragnar, M.D. (PM contractor, Environmental Hygienist, U of Gothenburg)
Professor of Environmental Hygiene, University of Gothenburg, Sweden. Worked on contract to Philip Morris overseeing biological laboratory work being performed at INBIFO, PM's biological labs in Cologne, Germany. PM paid Rylander $150,000 per year (salary determined from Bliley PM doc 2022850392, from 1992)
- Region
- Switzerland
- Germany
- Type
- SCRT, REPORT, SCIENTIFIC
- BIBL, BIBLIOGRAPHY
- CHAR, CHART, GRAPH, TABLE, MAPS
- SPEC, SPECIFICATION
- Litigation
- Stmn/Produced
- Named Person
- Angelljames
- Alarie
- Daly, D.
- Gearhart
- Henning
- Jaeger
- Kratschmer
- Zitnik
- Reininghaus, Wolf (INBIFO, Gen. Mgr., Contract Research)
Institut fur Biologische Forschung, Cologne, Germany - (Philip Morris' offshore research lab).
- Pruhs, D. - Biologist, INBIFO
- Teredesai, A. (Pathologist, INBIFO, c. 1987)
- Romer, Ewald (Quality Assurance Biologist, INBIFO, c. 1987)
- Operation/Project
- Secondhand smoke biological testing
- Named Organization
- Antiseptica Chemisch Pharmazeutische Pro
- Baker Chemikalien
- Bbc Goerz
- Beckman Instruments
- Braun Melsungen
- Brunning Und Frevel
- Drager
- E Merck
- Ehret
- Faust
- Filtrona Filter
- German Comm of Standards
- H Borgwaldt
- H Eggersmann
- Henkel
- Hewlett Packard
- INBIFO, Intitut Fur Biologische Forschung (Philip Morris' secret biological research lab in Europe)
"INBIFO" stands for Institut Fur Biologische Forschung, or Institute for Biological Research. It is located in Germany. Philip Morris acquired Inbifo on June 30, 1971. Its stated mission was "quantitative biological product evaluation" by using "comprehensive toxicological and physiological testing. Major activities are listed as: product evaluation and modifications, product ingredients and ETS-related technical knowledge and smoke components. Inhalation toxicology was a key feature of Inbifo. (Derived from Bates No. 2505235055/5088)
- Infors
- J Wolfe
- Kipp + Zonen
- Leybold Heraeus
- Linde
- Maihak
- Makrolon
- Messer Griesheim
- Metrawatt
- Miele
- Millipore
- Perkin Elmer
- Rota
- Sartorius
- Serva Feinbiochemica
- Sigma Chemie
- Team Aminal Treatment
- Team Analytical Chemistry
- Team Inhalation
- Unilife
- Wheaton Scientific
- Yellow Springs Instrument
- A Sauter
- Subject
- secondhand smoke
- Secondhand Smoke/Constituents
- secondhand smoke/health effects
- Secondhand Smoke/Toxicity
Document Images
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ao~=~O-PA
DR. MED. R. RYLANDER
c/o FABRIQUES DE TABAC DE REUNIES S.A. DRRS/UN
UN40 (R) Al
SWITZERLAND
COPY NO.:
R E P 0 R T P- 0500/3104
ACUTE INHALATION STUDY WITH
SIDESTREAM CIGARETTE SMOKE OF
STANDARD REFERENCE CIGARETTE 2R1 AND
AMMONIA VAPOR
ON RATS
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1NB1FO Institut fiir biologische Forschung Koln
REPORT P 0500/3104 UN44 (R) B23 - PAGE 0-1
CONTENTS PAGE
ABBREVIATIONS 0-7
1 SUMMARY 1-1
1.1 Objective 1-1
1.2 Principle 1-1
1.3 Experimental 1-2
1.3.1 Grouping, dosing and measurements 1-2
1.3.2 Generation and monitoring of test substances 1-3
1.3.3 Exposure 1-4
1.4 Results 1-4
1.4.1 Exposure to diluted sidestream smoke 1-4
1.4.2 Exposure to pure ammonia vapor 1-5
1.4.3 Conclusions 1-6
TABLE A: IRRITATION INDEX OF GAS/VAPOR PHASE COM-
PONENTS IN MICE
1-8
2 RESPONSIBILITY 2-1
2.1 Project Management = 2-1
2.2 Contributing Teams 2-1
3 QUALITY ASSURANCE STATEMENT 3-1
4 INTRODUCTION 4-1
5 TEST SUBSTANCES 5-1 .)
~
0
5.1 Cigarettes 5-1 cn
~
5.2
Ammonia
5-6 N)
~
~
TABLE B: SPECIFICATION OF STANDARD REFERENCE CIGARETTE
TYPE 2R1
5-2 T
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1NB]FO lnstitut fiir biologische Forschung Koln
REPORT P 0500/3104 IIN44 (R) B24 PAGE 0-2
CONTENTS (continued) PAGE
TABLE C: PHYSICAL PROPERTIES AND CHEMICAL COMPOSITION
OF FILLER OF STANDARD REFERENCE CIGARETTE
TYPE 2R1
5-3
TABLE D: SMOKE COMPONENTS OF STANDARD REFERENCE
CIGARETTE TYPE 2R1 5-5
6 METHOD
6.1 Chronology 6-1
6.2 SPF Conditions 6-1
6.2.1 Area for which SPF conditions are
obligatory 6-1
6.2.2 General working conditions 6-1
6.2.3 Entrace to the SPF Laboratory 6-1
6.2.4 Chemical disinfection 6-14
6.3 7%nimals and Animal Treatment 6-15
6.3.1 Animals 6-15
6.3.2 ~
Animal housing
6-16
6.3.3 biet and drinking water 6-18
6.4 Inhalation 6-21
6.4.1 Production of cigarette sidestream smoke 6-21
6.4.2 Production of ammonia vapor 6-24
6.5 Groups and Doses 6.28
6.6 Determination of Cigarette Parameters and
Smoke Components 6-29
6.6.1 Determination of cigarette weight 6-29
6.6.2 Determination of static burning rate 6-29
6
6
3 Determination of resistance to draw r")
6-30 ~
.
. 0
~
6.6.4 Determination of puff number 6-31 cn
~
6.7 Determination of Total Particulate Matter (TPM)
~
Concentration 6.32 ~
6.8 Determination of Carbon Monoxide Concentration 6-33
zze.
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IN81FO {nstitut fur biologische Forsctlung KbIn
REPORT P 0500/3104 UN44 (R) B25 PAGE 0-3
CONTENTS (continued) PAGE
6.9 Determination of Nicotine Concentration in
Particle Phase 6-34
6.10 Determination of Ammonia (NH3) Concentration 6-37
6.11 Biological Observations and Determinations 6-41
6.11.1 General condition, behavior and mortality 6-41
6.11.2 Determination of body weight 6-41
6.11.3 Determination of food consumption 6-44
6.11.4 Determination of respiratory frequency 6-45
6.11.5 Determination of rectal temperature 6-46
6.11.6 Determination of CO2 production of rats 6-47
TABLE E: GROUPS, EXPOSURE AND DOSES 6-27
TABLE F: CHECKLIST "SIGNS OF INTOXICATION, RAT" 6-42
FIGURE A.1: CHRONOLOGY, 0-GR 6-2
FIGURE A.2: CHRONOLOGY, 1.1-GR 6-3
FIGURE A.3: CHRONOLOGY, 2.1-GR 6-4
FIGURE A.4: CHRONOLOGY, 3.1-GR 6-5
FIGURE A.5: CHRONOLOGY, 4.1-GR 6-6 . ::.
FIGURE A.6: CHRONOLOGY, 5.1-GR 6-7
FIGURE A.7: CHRONOLOGY, 1.2-GR 6-8
FIGURE A.8: CHRONOLOGY, 2.2-GR 6-9
FIGURE A.9: CHRONOLOGY, 3.2-GR 6-10
FIGURE A.10: CHRONOLOGY, 4.2-GR 6-11
r.
FIGURE A.11: CHRONOLOGY, 5.2-GR
- 6-12 cn
G
~
~
FIGURE B: DIET CONTAINER 6-20
~
~
FIGURE C:
FIGURE D:
FIGURE E: SIDESTREAM SMOKING EQUIPMENT 6-22
AMMONIA EVAPORATION AND EXPOSURE EQUIPMENT 6-25
IMPACTION TRAP 6-39 m
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INBIFO lnstitut fiic biologische Forschung K31n
REPORT P 0500/3104 UN44 (R) B26 PAGE 0-4
CONTENTS (continued)
7 STORAGE OF RECORDS
8 RESULTS
8.1 Text
8.1.1 Specification of used cigarette type 2R1
8.1.2 Concentration of inhalant components
8.1.3 M~rtality
8. 1.~-G neral ond~i~ion and behavior
8.1.jr Body weight
8.1.9- Food consumption
8.1.f Body temperature
8.1.? Respiratory frequency
8.1.94~ Carbon dioxide exhalation
8.2 Tables and Figures
SPECIFICATIONS OF CIGARETTE, TYPE 2R1
CONCENTRATION OF INHALANT COMPONENTS
SYNOPSIS, GROSS PATHOLOGICAL FINDINGS
GENERAL CONDITION AND BEHAVIOR
BODY WEIGHT, MEANS AND RELATIVE STANDARD DEVIATIONS
DECREASE OF BODY WEIGHT
FOOD CONSUMPTION, MEANS AND RELATIVE STANDARD
DEVIATIONS
DECREASE OF FOOD CONSUMPTION
MEAN VALUES OF BODY TEMPERATURE
PAGE
7-1
8-1
TABLE
1
2
3
4 "~
~
5 a
~
~
6 PQ
~
~
7 o
8
9.1 TO 9.10
BODY TEMPERATURE 10
DECREASE OF BODY TEMPERATURE 11
MEAN VALUES OF RESPIRATORY RATE 12.1 TO 12.11
REPIRATORY FREQUENCY 13
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(NB1FO institut fiir bialogische Forschung Kaln
REPORT P 0500/3104 UN44 (R) B27 PAGE 0-5
CONTENTS (continued) TABLE
DECREASE OF RESPIRATORY FREQUENCY 14
CARBON DIOXIDE EXHALATION OF SHAM-EXPOSED RATS 15
CARBON DIOXIDE EXHALATION OF SIDESTREAM SMOKE
EXPOSED RATS
16
CARBON DIOXIDE EXHALATION OF AMMONIA EXPOSED RATS 17
C02 EXHALATION 18
DECREASE OF C02 EXHALATION 19
FIGURE
RELATIONSHIP BETWEEN TPM AND AMMONIA CONCENTRATION
IN DILUTED SIDESTREAM SMOKE
BODY WEIGHT DEVELOPMENT, SIDESTREAM SMOKE-EXPOSED
RATS
i
2
BODY WEIGHT DEVELOPMENT, PURE AMMONIA-EXPOSED RATS 3
RELATIONSHIP BETWEEN AMMONIA CONCENTRATION AND BODY
WEIGHT DECREASE
4
FOOD CONSUMPTION, SIDESTREAM SMOKE-EXPOSED RATS 5
FOOD CONSUMPTION, PURE AMMONIA-EXPOSED RATS G 6
RELATIONSHIP BETWEEN AMMONIA CONCENTRATION AND DE'REASE ~
OF FOOD CONSUMPTION 7 Q
~
BODY TEMPERATURE DURING EXPOSURE, SIDESTREAM SMOKE- Ui .
~
EXPOSED RATS 8 ~
0
0
BODY TEMPERATURE DURING EXPOSURE, PURE AMMONIA-EXPOSED
RATS
9
RELATIONSHIP BETWEEN AMMONIA CONCENTRATION AND
DECREASE OF BODY TEMPERATURE
10
RESPIRATORY RATE DURING EXPOSURE, SIDESTREAM SMOKE-
EXPOSED RATS
11
RESPIRATORY RATE DURING EXPOSURE, PURE AMMONIA-EXPOSED
RATS
12
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INBIFO Institut fur biologische Forschung Koln
REPORT P 0500/3104 UN44 (R) B28 PAGE 0-6
CONTENTS (continued) FIGURE
RELATIONSHIP BETWEEN AMMONIA CONCENTRATION AND
RESPIRATORY FREQUENCY
13
CARBON DIOXIDE EXHALATION DURING EXPOSURE, SIDESTREAM
SMOKE-EXPOSED RATS
14
CARBON DIOXIDE EXHALATION DURING EXPOSURE, PURE
AMMONIA-EXPOSED RATS
15
RELATIONSHIP BETWEEN AMMONIA CONCENTRATION AND
DECREASE OF CARBON DIOXIDE EXHALATION
16
PAGE
9 REFERENCES 9-1
I
~
This report,including title page, contains '. ages.
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INBIFO Institut fur biologische Forschung Koin
REPORT P 0500/3104 tJN40 (R) A5
PAGE 0-7
ABBREVIATIONS (a,b)
AC : team Analytical Chemistry
AT : team Animal Treatment
DIN : German Committee of Standards (Deutsches Institut
Ex
ISH : SH-index, thiol reactivity of cigarette smoke
L : 1 ight =
LD : light/darkness
M : mean
N : number of individual values
n. a. d.: nothing abnormal discovered
r : correlation coefficient
RT : room temperature
RSD : relative standard deviation
SOP : standard operating procedure
SPF : specific pathogen free
ST : standard time
TPM : total particulate matter
x g : centrifugal force in terms of the constant of
gravitation (1 x g = 9.81 m/s2)
fur Normung)
: x as exponent to the base 10, e. g. E2 = 102
H : team Inhalation
(a) in addition to those, which are explained immediately on the
same page
(b) Units are given in accordance with SI-norms (Systeme
International d'Unites).
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INBIFO Institut fur biologische Forschung Kaln
REPORT P 0500/3104 UN40 (R) A6 PAGE 1-1
1 SUMMARY
1.1 Objective
i n i'p_" ~ ~-C~
~
In this acute inhalation study the inluence of various concentra-
tions of SIDESTREAM CIGARETTE SMOKE and of AMMONIA VAPOR on the
body weight development, the food consumption, the respiratory
frequency, the carbondioxide production and the rectal temperature
of head-only-exposed rats and the correlation between these
parameters was investigated.
1.2 Principle
The aim of this 1-day inhalation study was to establish a dose=-
response relationship for the irritative capacity of_ sidestream
cigarette smoke in comparison to pure ammonia vapor, which is
supposed to be 1 of the main irritative factors in sidestream
smoke. Additionally, this study serves as a range finding study to
future 21-day inhalation studies with the same test compounds.
Accordingly, the applied sidestream doses covered the range
between the no-effect level and a sidestream dose, which was
2 times higher than the maximally tolerated daily sidestream total
particulate matter (TPM) dose applicable in 21-day studies.
Rats were kept in exposure tubes of a head-only exposure chamber.
They were exposed for 7 hours to a constant concentration of
sidestream smoke of standard reference cigarette 2R1 or to
pure ammonia vapor. During exposure the respiratory frequency or
the recta~~mperature as well as the carbon4_dioxide production of
the rats w-as continuously recorded.
The relativ erence between the respiratory frequency, ~
the CO roduction and rectal temperature before, during and
.`b0.1592~03
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INBIFO Institut fur biologische Forschung K6In <!5
REPORT P 0500/3104 UN40 (R) A7 PAGE 1-2
Wi^t-
agter- test substance exposure Va-s evaluated as a function of the
test substance concentration.
Sidestream smoke was produced in a 30-port INBIFO smoking machine,
appropriately diluted and dynamically flown through a 24-place
exposure chamber. Ammonia vapor was produced by dosing a constant
flow of an aqueous solution of NH40I-,T~, into a stainless steel
iw.n
cv4~VLGl.VL, k.:Viuiec:Leu w au ex~.rvSULe uuaiiw
r
11,Li
ll
The highest concentration of sidestream smoke co tained 0.25
milligrams TPM/liter and approx. 46 micrograms ammonia/liter.
Lower sidestream and higher 'ammonia concentrations were chosen
from, a geometrical series withqconstant factor of 2 between
concentrations. Concentrations below the apparent no-effect level
were not administered. Each dose level of each test substance was
assayed with 1 dose group of 20 male Sprague Dawley rats.
1.3 Experimental
1.3.1 Grouping, dosinq-and-measurements
---- -----
Male Sprague Dawley rats of approx. 260 grams body weight were
allocated randomly to 11 experimental groups: 5 of these groups
were exposed to sidestream smoke of TPM concentrations of 0.013,
0.025, 0.055, 0.12 and 0.25 milligrams/liter respectively,
5 groups were exposed to pure ammonia vapor concentrations
of 31.8, 50.7, 11?4, 213A and 411j-- t micrograms/liter.
Immediately before and after the 7-hour exposure all rats were
sham-exposed for a period of 30 minutes and 60 minutes respec-
tively.
1 additional group of 10 rats was sham-exposed for 8.5 hours and
served as a control group.
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