FORENSIC SCIENCE PROJECT

REPORT ON:

CARBON MONOXIDE POISONING

Submitted to: Submitted by:

Dr.Ajay Ranga Sukhanpreet Kaur Rangi

Roll No. 55/15

Semester – 9th

Section – A

1
 STUDENT CERTIFICATE

University institute of legal studies , Panjab university , Chandigarh

CERTIFICATE

This is to certifiy that Sukhanpreet Kaur Rangi , a student of 9th semester of

B.A.LLB(Hons) has successfully completed the research on the project of forensic
science related to (carbon monoxide poisoning ) under the guidance of Dr. Ajay
Ranga during the year 2019-2020

Name of student Name of teacher

Sukhanpreet Kaur Rangi, Dr.Ajay Ranga

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 SUPERVISOR CERTIFICATE

This is to certify that the work incorporated in the project titled “carbon monoxide
poisoning’’ is a research work carried out by Sukhanpreet Kaur Rangi, Roll no.
55/15, 9th semester, B.A.LLB(Hons)

Under my guidance and supervision for the award of internal assessment for 9th
semester in the year 2019- 2020.

To the best of my knowledge and belief that the project report:-

1. Embodies the work of the candidate herself

2. Has duely been completed
3. Fulfills the requirements of the internal assessment
4. Is upto the desired standard both in the respect of the contents and language
for being referred to the examination

Sign

----------------------------------------

Dr. Ajay Ranga

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 TABLE OF CONTENTS

Meaning And Importance Of 5

Toxicology In Forensic Science

Toxicological Examination In Gas- 5

Related Poisoning Incidents

CARBON MONOXIDE (CO)

 Properties: 6
 Sources 6
 Reaction with blood and how it 6
works
 Symptoms 8

Detecting CO, Identification and

quantification of CO
 Test for identification purposes 9
 Test for quantification purposes

Study conducted and Cases reported

all in various countries
 India 10
 Japan 15
 Portugal 15
 Denmark 16

Observation 17

Conclusion 18

Webliography 19

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 Meaning And Importance Of Toxicology In Forensic Science

Toxicology is a scientific discipline, overlapping with biology, chemistry,

pharmacology, and medicine, that involves the study of the adverse effects of
chemical substances on living organisms and the practice of diagnosing and
treating exposures to toxins and toxicants1

Toxicological examination in forensic practice is important for the proper

diagnosis of acute poisoning . The forensic pathologist requests toxicological
analysis to forensic toxicologist in case of poisoning or poisoning suspected cases.
It will usually consist of two-stage testing at autopsy. The first step is usually
performed as a screening test. The second step is required for the identification and
quantification of its causative agent following the confirmatory test.

Toxic gaseous substances are one of the targets for toxicological examination in a
daily forensic practice. Simplified analysis of gaseous substances involves both
first and second step of toxicological examination in forensic practice.

Toxicological Examination In Gas-Related Poisoning Incidents

Gaseous substances can cause acute poisoning. They get absorbed into the body by
inhalation. Most of them do not produce specific symptoms but they can induce
dizziness, lethargy, headaches, and suffocation. There is no specific finding at
autopsy in most poisoning cases . The presence of gaseous poisons is usually
indicated by the circumstances of the incident, and involvement of gaseous
substances is sometimes indicated by circumstantial evidence . in this project

1
http://www.jfmpc.com/article.asp?issn=2249- last visited 20th October

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report we will discuss about the detection ,affects ,and analysis of deaths caused by
carbon monoxide poisoning 2

CARBON MONOXIDE (CO)

Properties:

Carbon monoxide (CO), which is one of the many ubiquitous contaminants of our
environment, is responsible for a large percentage of the accidental poisonings and
deaths reported throughout the world each year.Carbon monoxide (CO) is a
colorless, odorless, tasteless, and nonirritating gas which makes it difficult for
those who are exposed, to detect it, Because CO lacks sensory warning properties,
it is commonly known as the “silent killer.” leading to unexpected death.

Sources:

CO is formed during an incomplete combustion of organic material, for example,

gasoline, coal, wood, propane, and natural gas. Common workplace sources of CO
include fuel-powered engines (motor vehicles, forklifts, generators, pumps, etc.),
fuel-burning heaters (furnaces, water heaters, boilers, space heaters, etc.), coke
ovens, and blast furnaces. However, the most common sources of CO are faulty,
poorly maintained, or inadequately ventilated gas appliances such as stoves and
heaters. Malfunctioning heating systems, improperly ventilated motor vehicles,
generators, grills, stoves, and residential fires may be listed in the common sources
of CO exposure

2 https://acutecaretesting.org/en/articles/postmortem-co-oximetry last visited 20th October

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Reaction with blood and how it works

CO has 210 times the affinity for hemoglobin than that of oxygen and impairs
oxygen delivery and peripheral utilization leading to cellular hypoxia. It can result
in a variety of acute symptoms in low doses, including headache, dizziness,
weakness, nausea, confusion, disorientation, and visual disturbances. In extreme
cases, exposure leads to unconsciousness, coma, convulsions, and even death. 3

CO is absorbed from the lung into the bloodstream. As the affinity of CO for
hemoglobin is 230–270 times greater than that of oxygen, it binds to hemoglobin
in erythrocyte, and forms carboxyhemoglobin (CO-Hb) .The formation of CO-Hb
(represented as a percentage of the total hemoglobin) in blood depends on various
factors such as the concentration of inspired CO, duration of CO exposure,
pulmonary ventilation, exercise, and health status. The toxicity of CO is thought to
be tissue hypoxia due to the formation of CO-Hb. Its binding is a reversible
process; however, as the binding between CO and hemoglobin is strong, the CO
elimination half-life is long, about 4–5 hours under room air ventilation for a
resting adult at sea level. The formation of CO-Hb decreases the capacity of
oxygen transport, and it causes insufficient oxygen supply in tissues .

The hypoxia due to CO-Hb formation causes signs and symptoms. Clinical
symptoms roughly correlate with CO-Hb levels (Table 1). The CO-Hb
concentration of nonsmoking healthy subjects is 1–3%, and around 5–8% in
smokers. No symptom is observed below 10% of CO-Hb levels. Neurological
symptoms such as headache, dizziness, nausea, and weakness are observed in CO-

3 https://www.intechopen.com/books/poisoning-from-specific-toxic-agents-to-novel-rapid-and-
simplified-techniques-for-analysis/simplified-analysis-of-toxic-gaseous-substance-in-forensic-practice-
experiences-from-japan

7
Hb level from 10 to 30%. Increase of respiration and heart rate, syncope, and
confusion are observed in 30–50% of CO-Hb level. When the level of CO-Hb
exceeds 50%, it becomes life-threatening. It is noteworthy that the value of CO-Hb
is important for the diagnosis of CO-poisoning or fire-related death. In addition to
hemoglobin, CO combines with heme-proteins such as myoglobin and cytochrome
oxidase, and it may cause the impairment of cardiac and neurological functions 4

Symptoms

Correlation of carboxyhemoglobin (CO-Hb) levels to clinical symptoms. Most of

gaseous substances cause little or no tissue damages. However, there will be
observed unique findings in some poisoning cases, such as cherry red appearance
of postmortem lividity in carbon monoxide poisoning,

The most characteristic appearance of the body in poisoning case is a cherry red
color of the skin. It is usually observed in cases where CO-Hb exceeds 30% . At
autopsy, the common findings include discoloration of blood, organs, and muscle
that become cherry red color, as a result of CO-Hb and carboxymyoglobin. Other
autopsy findings such as pulmonary edema and generalized organ congestion are
also observed .

CO-Hb (%) Clinical symptom

0–10 No symptom

10–20 Headache, ear ringing, fatigue

20–30 Headache, weakness, nausea, vomiting

4 https://www.ncbi.nlm.nih.gov/pubmed/24794841

8
30–40 Severe headache, dizziness, nausea, vomiting

40–50 Syncope, confusion, increased respiration and heart rate, muscle

weakness
50–60 Coma, convulsions, depressed respiration

60–70 Coma, convulsions, cardiorespiratory depression, often fatal

70+ Respiratory failure, death

Detecting CO, Identification and quantification of CO

With regard to the identification and quantification of CO, several methods and
techniques have been reported . The CO-Hb is relatively stable under storage in
cool and dark conditions .It is important to note that postmortem production of CO
has been reported in some conditions, and therefore, it is recommended not to use
body cavity fluids such as pleural effusion for the measurement of CO in severe
putrefied case .

Test for identification purposes- the qualitative test for CO includes color test
and microdiffusion tests.

Color test is a simple procedure where a blood sample mixed with 0.01 M
ammonia solution (1:20) or a few drops of blood are added to some 10% sodium
hydroxide solution .This test is based on the fact that CO-Hb is relatively tolerant
to alkaline condition. However, as other simple methods have been established,
color testing for CO poisoning is now rarely required and not recommended

The microdiffusion test using Conway cell or on the filter paper have been
reported. It is based on reaction with palladium chloride. This is still the most
widely used method since it was invented by Conway in 1944.

Test With regard to quantification,- spectrophotometric method, gas

chromatography, detection tube, and oximeter are used. 5

5
http://www.jfmpc.com/article.asp?issn=2249-

9
The spectrophotometric method is the most popular, and various assay procedures
have been reported. CO-Hb could be determined by the changes of absorption
spectrum in either Soret (410–425 nm) or visible region (500–600 nm) .In our
laboratories, we perform the measurement of the spectrum of blood sample by
adding sodium hydrosulfite. The addition of sodium dithionite reduces
oxyhemoglobin without affecting the CO-Hb. This procedure is simple, and it does
not need an extraction from the sample. Figure shows the spectra of blood samples
from a normal nonsmoker (CO-Hb: 0%) and CO poisoning victim (CO-Hb: 68 and
95%, respectively). Twin-peaked spectrum was observed in CO poisoning sample.

Spectra of blood sample from CO-Hb: 0, 68, and 95%, respectively.

Gas Chromatography - The CO is extracted and introduced in gas

chromatography. Various methods and apparatus have been reported for its
extraction . And the released CO is detected by the thermal conductivity detector
(TCD) or the flame ionization detector with the catalytic reduction of the CO to
methane. As this method is a direct measurement of CO contents in the sample as
well as a measurement of the hemoglobin, two measures represent the percentage
of CO-Hb. Application of gas chromatography equipped with semiconductor
detector has been reported for forensic practice . This gas chromatography system
(sensor gas chromatography, sGC) is highly sensitive for CO and has some
advantages such as portability and easy handling. This apparatus does not need a
gas cylinder as it uses the room air as the career gas. Although it is not commonly
in use, further application in the field of forensic medicine would be expected.

The detector tube method is widely used for the determination of various gaseous
substances. It is also applied for the quantitation of CO in blood. This apparatus
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consists of a CO-separator tube, CO-detector tube, and aspirating pump. The CO-
separator tube is packed with silica gel particles coated with ferricyanide .The CO-
detector tube is packed with silica gel particles coated with sulfite palladium
potassium .These tubes and pump are connected in series. The CO in blood is
released following the injection of blood sample (200 μL) in CO-separator tube,
and the released CO gas is detected by the CO-detector tube, followed by the
aspirating of the pump. As the detector tube is easy to carry at the scene where an
incident has taken place or to a point-of-care testing, it is applied to not only
screening test, but also for quantitation.

Oximeter is routinely used for laboratory test , and it is also applied in forensic
medicines . This instrument uses seven wavelengths in the visible region for the
determination of various hemoglobin species, such as oxyhemoglobin, CO-Hb,
reduced hemoglobin, and methemoglobin. It automatically analyzes the proportion
of each species of hemoglobin and oxygen contents. This oximeter system
requires 50 μL of blood for a single measurement, and it may be a valid option in
case of difficult blood sampling due to severe blood loss. As there are many
advantages such as no necessity of sample preparation, easy handling, and
portability, it is suitable for forensic practice. In a recent study, it has been reported
that squeezed splenic blood can be used as an alternative specimen for CO-Hb
measurement using oximeter .

6
Portable oximeter (AVOX 4000) and its operation. Sample cartridge is shown in lower left-hand corner.

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Portable oximeter (AVOX 4000) and its operation. Sample cartridge is shown in
lower left-hand corner.

Study conducted and Cases reported all in various countries

INDIA

Few studies on CO poisoning and only a few cases of fatal CO poisoning with
different scenarios have been reported from India, but no article has shed light on
the preventive aspect of it. Considering the public health importance of the CO
poisoning and the knowledge gap this study was undertaken to see the pattern of
fatal CO poisoning and to discuss preventive aspect.

Materials and Methods: It was a retrospective descriptive study of fatal CO cases

which were autopsied at All India Institute of Medical Sciences, New Delhi, from
the year 2010 to the year 2015. Only those cases were included in this study in
which death was opined due to CO poisoning, based on characteristics autopsy
findings, toxicology report from forensic science laboratory, and crime scene
investigation report, and after ruling out other causes of death. The cases were
analyzed as per age groups, circumstances of death, season of death, and sources of
CO formation. Results and Discussion: The study involved 40 cases of fatal CO
poisoning. About 80% of cases were reported in winter months. The maximum
cases were reported in the month of January followed by November and
December. All the cases except one, died with a source of CO nearby and the
person was inside a room or some closed space without ventilation. Source of CO
was firepot and electric room heater in most of the cases. Some cases were of CO
build inside the car with a running engine. Most of the cases occurred
accidentally.
Conclusion: Clustering of cases is seen in winters. Poisoning can occur in different
ways. The study documents the various possibilities of CO poisoning and
advocates community education targeting the high-risk groups and masses,
especially during the winter season. 7
Result
The study involved 33 incidences of CO poisoning with 40 victims reported at All

7
Asit Kumar SIkarey, Sumit Dixit “fatal carbon monoxide poisoning; a lesson from a retrospective study” vol 6,
journal of family medicine and primary care, 2017

12
India Institute of Medical Sciences, New Delhi. All the cases were brought dead to
the casualty of the institute. Out of them, 35 were males and 5 were females. Age
of the victims was varying from 8 to 59 years with maximum number of cases seen
in the age group 31–45 years followed by 15–30 years, and all of the cases, except
one, were from economically productive age group [Table 1].

Ninety percent of the incidents were reported in winter months, i.e., November,
December, January, and February month of the year. Incidents were about equally
distributed among November, December, and January month of the year [Table 1].
8
All the victims, except one, died with a source of CO nearby and the person was
inside a room or some closed space without any ventilation. In one case, the
deceased person was sleeping in a field inside an impervious mosquito net with a
firepot inside, which again created a zone of high CO concentration. Most of the
victims were alone at the time of the incident, except in four incidents, there were
more than one victim, and in one case, there was a stray dog that also died sleeping

8
Asit Kumar SIkarey, Sumit Dixit “fatal carbon monoxide poisoning; a lesson from a retrospective study” vol 6,
journal of family medicine and primary care, 2017

13
in the room with a security guard. Source of CO was firepot in most of the cases.
Two scenarios of death were of CO build inside the car with the engine and car
blower running. Most of the incidents occurred accidentally, except one where a
person used CO cylinder to poison himself, purchasing it online on the pretext of
research [Table 2]. 9

Except in four incidents, victims were daily wagers, servants, or security guards
who used to sleep in a single room accommodation or in the security booth.
Among those four incidents, in one incident, the family was involved with two
women, one boy, and one girl, succumbing to CO poisoning when their house
burnt. In the second incident, three students were found dead inside a car with
running engine. In the third incident, a businessman was found dead inside his car

9
Asit Kumar SIkarey, Sumit Dixit “fatal carbon monoxide poisoning; a lesson from a retrospective study” vol 6,
journal of family medicine and primary care, 2017

14
with running engine, and in the fourth incident, a student committed suicide using
CO cylinders

JAPAN

CO is the leading cause of poisoning death in Japan ,and also a common cause of
poisoning in the United States.The annual number of victims by CO poisoning is
about 2000–4000 in Japan, including accidental or suicidal cases .10

PORTUGAL

Study conducted in portugal Faculty of Medicine, University of Coimbra,

Coimbra, Portugal11

This study presents the epidemiology and the postmortem forensic aspects in cases
with a carboxihemoglobin (COHb) analysis, from autopsies performed at the
Forensic Pathology Department of the Centre Branch of the National Institute of
Legal Medicine and Forensic Sciences of Portugal. Between January 2000 and
December 2010, 69 COHb analyses were requested in our institution. In
approximately 70% of the situations, circumstantial information included a Carbon
Monoxide (CO) source at the death scene. More than half of the cases presented
thermal lesions, cherry-red lividity, and cherry-red blood and viscera coloration
were found in, approximately, 30% of the cases. Fourteen cases were recorded as
CO poisonings. The highest number of poisonings occurred in 2000, with most of
the cases in winter (53.8%), in 51-60 years-old male individuals. 69.2% of the
poisonings were accidental and the remainder were suicides, being fires the most
frequent sources of CO (38.5%). Cherry-red lividity was present in 61.5% of the

10 Hiroshi Kinoshita, Naoko Tanaka, Ayaka Takakura, Mostofa Jamal, Asuka Ito, Mitsuru Kumihashi,
Shoji Kimura, Kunihiko Tsutsui, Shuji Matsubara and Kiyoshi Ameno, ‘Simplified Analysis of Toxic
Gaseous Substance in Forensic Practice: Experiences from Japan’ DOI:
10.5772/intechopen.70029Published: December 20th 2017

11 Ruas F, Mendonça MC, Real FC, Vieira DN, Teixeira HM, ‘Carbon monoxide poisoning as a cause of
death and differential diagnosis in the forensic practice: a retrospective study, 2000-2010.” J Forensic
Leg Med. 2014 May;24:1-6. doi: 10.1016/j.jflm.2014.02.002. Epub 2014 Feb 12.

15
cases, and all of them presented cherry-red blood and viscera coloration. Older
individuals and those with thermal lesions presented lower COHb levels, and
politrauma was the most frequent cause of death among the negative cases. It is
possible to conclude that the forensic aspects of CO poisonings interact in a
complex way, and differential diagnosis is not straightforward. This study also
emphasizes the role played by public prevention campaigns and improvement of
heating appliances in reducing the number of accidental CO poisonings, and the
importance of preventing urban and forest fires, the major source of CO among us

DENMARK

Forensic aspects of carbon monoxide poisoning by charcoal burning in Denmark,

2008-2012: an autopsy based study was conducted. By Prof .Nielsen , Gheorghe
A, Lynnerup N from Department of Forensic Medicine, Faculty of Health
Sciences, University of Copenhagen

The study showcased that Carbon monoxide (CO) inhalation is a well-known

method of committing suicide. There has been a drastic increase in suicide by
inhalation of CO, produced from burning charcoal, in some parts of Asia, and a
few studies have reported an increased number of these deaths in Europe. CO-
related deaths caused by charcoal burning have, to our knowledge, not been
recorded in the Danish population before. In this retrospective study we present all
autopsied cases of CO poisoning caused by charcoal burning in the period 2008-
2012. 19 autopsied cases were identified, comprising 11 suicides, 4 accidents, and
2 cases of maternal/paternal filicide-suicide. The mean age of decedents was
38.2 years and the majority of the decedents were men. In 16 cases
carboxyhemoglobin levels were above 50 % and in 14 cases we found distinctive
cherry red livor mortis. Various concentrations of ethanol and drugs were found in
9 cases. Data suggest that this method of death has increased significantly in
Denmark. Therefore, it is highly relevant to draw attention to the subject, to
increase awareness as well as prevent future escalation.12

By Prof .Nielsen , Gheorghe A, Lynnerup N ‘Forensic aspects of carbon

12

monoxide poisoning by charcoal burning in Denmark, 2008-2012: an autopsy

based study’et al. Forensic Sci Med Pathol. 2014
16
 OBSERVATION

This study was undertaken to study the pattern and circumstances of CO poisoning
death and to discuss its preventive aspect. Patterns of the high number of cases of
CO poisoning in the colder months of November to February and a less number of
cases in the hotter months of June to August is seen in this study in concordance
with other studies. Deaths were among economically productive age group and
more commonly accidental as in other studies worldwide, but CO has long been
recognized as a means of deliberate suicide too, as in our study.

Firepots, car engines, and room heaters were sources of CO in this study as well as
in other studies. Use of room heaters, which is shown to be one of the top sources
of CO fatalities, is predominant during the winter season and can release high
amounts of CO if the equipment is not functioning properly or the place is not
ventilated. In this study too, the rooms in which the deceased were sleeping with
the firepots/room heater were having poor ventilation with vents sealed to prevent
any,cold-breeze.

This study along with other case reports from India and worldwide showed that the
main cause of CO poisoning is accidental collection of CO in a non ventilated
room/closed space. Detection of CO collection is difficult for sleeping person
because of its “silent” physical and chemical property. People should be alarmed
about the danger of CO collection and poisoning in a nonventilated room while
using various room warming mechanisms such as firepots and room heater and by
putting warning labels on various CO-producing appliances. They should be taught
to avoid using these appliances without proper ventilation in the area to avoid
collection of dangerous CO. Alternatively, a CO alarm can be put in the house and
closed area, even in cars, while using these appliances and it should be promoted
as smoke alarms. This prevention is important because it takes toll among
economically productive age groups.

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 CONCLUSION

CO poisoning mostly occurs during winter season as evident. There is the use of
various room heating mechanisms during this period which leads to the formation
of CO gas in closed non ventilated room. Special efforts should be done during this
period to educate the masses regarding this silent killer, and various preventive
measures should be taken with which morbidity and mortality can be minimized.
High-risk groups, such as people living in single room accommodation, using
heater, blower, firepot, especially in winters, as found in this study, should be
identified and they should be especially warned and educated regarding such risk.

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 Webliography

 https://www.ncbi.nlm.nih.gov/pubmed/24794841

 https://www.ncbi.nlm.nih.gov/pubmed/25002407

 http://www.jfmpc.com/article.asp?issn=2249-

4863;year=2017;volume=6;issue=4;spage=791;epage=794;aulast=Sikary

 https://www.intechopen.com/books/poisoning-from-specific-toxic-agents-to-

novel-rapid-and-simplified-techniques-for-analysis/simplified-analysis-of-

toxic-gaseous-substance-in-forensic-practice-experiences-from-japan

 https://acutecaretesting.org/en/articles/postmortem-co-oximetry

19