Retrospective Observational Study of Organophosphate

Poisoning Trends in Uttar Pradesh

A DISSERTATION SUBMITTED TO

UNIVERSITY OF
LUCKNOW
FOR THE PARTIAL FULFILLMENT FOR THE AWARD OF THE
DEGREE OF

M.Sc. Forensic Science

(SESSION 2021-23)
[Specialization in Forensic Biology]
SUBMITTED TO:
Dr. KEYA PANDEY
(Head of the Department)

BY

Krishn Praddhumn

ROLL NO.2110014205012

M.Sc. Forensic science

Department of Anthropology

i
 DECLARATION

I hereby declare, that the dissertation entitled “Retrospective Observational

Study of Organophosphate Poisoning Trends in Uttar Pradesh” submitted by me
in the partial fulfilment for the M.Sc. in Forensic Science in the Department of
Anthropology, University of Lucknow is an outcome of my original work and
efforts under the supervision of Dr. Keya Pandey, HOD of The Department of
Anthropology, University of Lucknow and Raj Kumar Sir, Head of Toxicology
Division at State Froensic Science Laborator, Lucknow, Uttar Pradesh. I hereby
confirm that this work has not been submitted in part or full to this or any other
university for the award of degree.

Krishn Praddhumn

2110014205012

ii
iii
 ACKNOWLEDGEMENTS
I deeply acknowledge Prof. Keya Pandey, Head, Department of
Anthropology, University of Lucknow, Lucknow for her constant support and
motivation for carrying this master’s dissertation.

I am very grateful to, Dr. Sudhir Kumar Director, SFSL, Lucknow, for providing
mean opportunity to work in this esteemed institution.

I am immensely grateful to Dr. Saurabh Pratap Singh Sir, Faculty in the

Department of Anthropology at the University of Lucknow, Lucknow, for
providing valuable academic assistance and consistently motivating me in the
successful completion of this dissertation.

I would like to convey my deep appreciation and thankfulness to RAJ KUMAR

sir, Scientific officer at SFSL, Lucknow, for being a constant source of
inspiration, guidance, and unwavering support throughout every phase of my
dissertation.

I extend my gratitude to various departments of SFSL and Department of

Anthropology, University of Lucknow, Lucknow, for their support at various
stages of my work.

I acknowledge the support of my parents, family, and friends for the

constant motivation and support for my dissertation work.

Krishn Praddhumn

M.Sc. Forensic Science

iv
 CONTENTS
S. No. Contents Page number

1. CERTIFICATE i

2. ACKNOWLEDGEMENTS ii

3. LIST OF TABLES iii

4. INTRODUCTION 1

5. REVIEW OF LITERATURE 19

6. METHODOLOGY 28

7. DATA COLLECTION 33

8. DATA ANALYSIS AND RESULT 36

9. CONCLUSIONS 41

10 REFERENCES 44

v
List of Tables:

Contents Description Page No.

Table 1. Rf values of some common OPCs 15

Table 2 Age Group 34

Table 3 Gender 34

Table 4 Residential Region 35

List of Figures:

Sno. Caption Page no.

1 Classification of Agriculture Poison 3
2 Classification Of OP compounds 4
3 A general Chemical structure of OP compound 9

4 Showing Mechanism of Action Of OP insecticides 10

List of Charts:

S. No. Chart Page no.

1 For Age Group 38
2 For Gender 39
3 For Geographical 40
Region

vi
 Section- First

Introduction

1
Organophosphates are the class of chemical compounds widely used in
agriculture, industry, and warfare and have played a significant role in
shaping the modern world. This adaptability has a price, as exposure to
organophosphates poses a serious public health risk. Organophosphate
poisonings, whether deliberate or unintentional, are caused by these
compounds' poisonous qualities, which interfere with the nervous system's
regular operation. Gaining a thorough understanding of the trends,
patterns, and consequences connected with organophosphate poisonings is
crucial in light of this continuous problem.

Agricultural poisons are the organic irritants used in agriculture field, they
are commonly known as pesticides. Pesticides are the poisonous
compounds that are commonly used to kill pests. Insects are the biggest
issues in the field of agriculture for a farmer, so they use insecticide to
overcome the issue. Pesticides are applied through aerial spray over the
plants and it get mixed with the plants through leaves, stems and soil and
hence roots without harming the plants.

When the insects come in the contact of the plant, the poison acts as
contact poison and absorbs through exoskeleton or when insect sits on the
plant, it consumes the poison along with the leaves. But it does not cause
any harm to human who consumes the grains, fruits or other parts of the
plants if used with recommended cautions. However, when it enters the
human body either through direct or indirect route, then it may cause
poisoning.

Pesticide poisoning is one of the biggest poisoning issues of rural world.

Organophosphate Poisoning is one of the most common poisoning cases in
emergencies at world wide. Ops inhibits CHE receptors of nervous system in

2
the body irreversibly even at lower concentration. Its clinical features may
include which is not limited to symptoms of Nausea and Vomitting which is
most common symptoms and miosis as most common sign. [1] Respiratory
failure is general cause of death in case of OP poisoning. OP is related with
death of estimate about 2,00,000 people every year. [2]

Classification of Agricultural Poisons

Insecticides Herbicides Fungicid

Rodenticides Miscellaneous
Agricultural Poisons

Organo- Carbamates
phosphorous
Pyrethroids
Organochlorin

Figure 1

OPs are generally used as insecticides, drain cleaners, rat killers, pesticides
and germicides.[3] All these products are cheap and have easy availability in
the markets and also does not require the checking of history and previous
records. In many developing countries like India people many a time fails to
keep such products securely at home or working places. Due to these faults

3
the products have easy accessibility and hence the product becomes poison
of choice.

Types of Organo-phosphorous
compounds

 Alkyl  Aryl  Others

a. Chlorpyriph - -Phorate
-Phosphamidon,
os Parathion
etc
b. Dimethoate -Diazinon,
c. Malathion etc
d. Monocrotoph
os
e. Quinolphos
f. TEPP, etc
Figure 2

OPs have the potential to kill in minutes of exposure. Due of social

degradation, unemployment, and poverty in developing nations, pesticide
products are frequently used for self-harm.[4] Suicidal poisoning with
household products (OPs, carbamates, pyrethrinoids, etc.) is the most
frequent type of poisoning, as per the data from the National Poison
Information Centre of India.[5] Death is influenced by the kind of substance
consumed, how much is consumed, how it is consumed, the person's health,
and diagnosis accuracy. Early diagnosis, the use of atropine in conjunction
with intravenous fluids, and appropriate respiratory support, including
oxygen therapy, all contribute to favorable outcomes.[6]

4
Many hundreds of patients poisoned by pesticides are treated annually in
hospitals in rural areas, with a case fatality rate of 15–35%.[7, 8].
Unfortunately, these medical organizations frequently lack the resources
and staff required to manage these critically ill patients; as a result, even
unconscious patients are managed on open wards. In rural Asia, better
medical treatment, the availability of antidotes and intensive care beds, as
well as restrictions on the most toxic pesticides, should lower the case
fatality rate for self-poisoning and significantly lower the number of suicide
fatalities.

OPs are acetylcholinesterase (AcH) inhibitors and they affects upon the
Acetylcholinesterase and cholinergic synapses. Although there is variation in
the clinical symptoms and indications depending on the nature of the
chemicals, amount eaten, intensity, time interval between exposure, and
hospital admission, respiratory failure and lung injury account for the
majority of patient deaths. [9]

In several countries in the Asia i.e Asian pacific region, South Asian region
(India, Sri lanka), self harm is a significant issue.[7] Self harm, in particular
self poisoning, is not merely used to take one's life; it can also be done to
attract attention, vent pain, or exact revenge.[10]

Fatal Dose [11]

The following compounds are extremely toxic (LD50: 1 to 50 mg/kg), or

highly toxic (LD50: 51 to 500 mg/kg)- Dichlorvos, Dimethoate, Methyl
Parathion, Mevinphos, Monocrotophos, Phosphamidon, Quinalphos, TEPP.

5
The following compounds are moderately toxic (LD50: 501 to 5000 mg/kg),
or slightly toxic (LD50: more than 5000 mg/kg)- Abate, Acephate, Malathion,
Phenthoate, Primiphos Methyl, Triazophos, and Trichlorphon.

Human fatal doses for these toxicants are less than 5 mg/kg, along with
arsenic (As), cyanide (CN-), and some others. As little as 2 mg of parathion
has been known to kill children.

Clinical Features Of OPP [13]

The typical clinical affects of OPP are following:

Muscarinic affects: [12]

The CNS has all five muscarinic receptor subtypes (M1-M5).

The heart, exocrine glands, and smooth muscles of the internal organs are
all innervated by postganglionic peripheral muscarinic receptors. The
sympathetic postganglionic fibres innervate the sweat gland.

Over Respiratory Tract: Increased bronchial secretions, bronchospasm, chest

tightness, dyspnoea, cough

Over Eyes: Blurred vision, conjunctival injection, dimness of vision, miosis

Over Gastrointestinal Tract: Cramping, diarrhea, nausea, vomiting

Over Urinary Tract: Incontinence

Over Cardiovascular System: Bradycardia, hypotension

Over Exocrine glands: Hyperamylesia, increased salivation

6
Nicotinic affects: [12]

Two categories of nicotinic receptors exist: periphery and central

The central nervous system (CNS) contains central nicotinic receptors Nn or

N2. Additionally, they are located in the adrenal medulla and sympathetic
and parasympathetic ganglia of the peripheral nervous system (PNS).

The level of the neuromuscular junctions is where peripheral nicotinic

receptors Nm or NI are found.

Muscle fasciculation, cramping, weakness, diaphragmatic paralysis,

respiratory failure, affect over Autonomic ganglia (tachycardia,
hypertension).

So we can say that OP poisoning can cause a patient to exhibit a wide range
of signs and symptoms. In many developing countries like India, generally
unconscious patients are frequently brought by their family, relatives and
neighbours who are unable to provide the information on the specific
poisonous substance or product to which the patient was exposed. In this
situation medical professionals/ doctors tries to identify the poisoning
through clinical features of the patients. With the help of clinical features
doctors may also determine about the seriousness of the poisoning which is
also important for treatment and legal matters in case of suicidal and
homicidal poisonings.

In the above situations, due to lack of information collected from the

relatives of the patients, many a times medical practitioners try to check the
clinical features of the patients. To confirm the doubtful poisoning in the
patients improper informnations given by relatives of patients, the blood,

7
urine samples or vomittus are collected as a sample which is to be sent for
confirmatory tests to the labs. Even in case the patient is died then the
postmortem examination is conducted by the medical officer, and on the
basis of the PM appearance he/she gives certain informations that helps in
legal perspective. This medicolegal examination also helps medical officer to
conclude the cause of death, many a times in case of poisoning for the
confirmation of causative substance that was related with death of person,
viscera is preserved and sent to the Forensic Science Laboratories for the
poison analysis through the legal channel (chain of custody).

There are many challenges for the forensic experts too for the analysis of
poisons from preserved visceral samples, blood samples, urine samples etc.
One such challenge they face is the contamination at the time of
preservation and sealing from medical centers or medico-legal centers. All
these things affect the results during poison analysis. They perform various
steps for analysis i.e extraction of poison, sample preparations for
instrumental techniques (Chromatographic techniques, Spectroscopic
techniques etc).

In cases of Pesticide poisoning viscera, serologically important samples are

preserved, in case of a patient are dead and in order to confirm the category
of pesticide and type of substance/product was consumed which is present
in the patient/deceased body viscera samples are sent to the Forensic
Science Laboratories. So for retrospective and observational studies the
major sources are critical care units of medical centres or emergency units,
yearly data from NCRB and Forensic science Laboratories.

This thesis embarks on a crucial exploration into the retrospective

observational study of organophosphate type poisonings trends that

8
registered in Uttar Pradesh during the selective period of time. By examining
historical data and trends, this research endeavors to shed light on the
epidemiology, clinical presentation, management, and outcomes of
organophosphate poisonings over time. The aim is not only to contribute to
the existing body of knowledge but also to provide insights that can inform
public health policies, prevention strategies, and clinical practice.

Figure 3 A general Chemical structure of OP compound

Action of OP compounds in Human Body

Pesticides containing organophosphorus are known to block esterase

enzymes, most notably acetylcholinesterase (AChE) in synapses and on the
membranes of red blood cells and butyrylcholinesterase (BChE) in plasma.
[14] Although acute butyrylcholinesterase inhibition does not appear to
generate clinical symptoms, acetylcholinesterase inhibition causes
acetylcholine to accumulate and acetylcholine receptors to be
overstimulated in synapses in the autonomic nervous system, central
nervous system, and neuromuscular junctions. [14] The diagnosis is based
on clinical suspicion, the distinctive clinical symptoms, the smell of

9
pesticides or solvents, and decreased butyrylcholinesterase or
acetylcholinesterase activity in the blood. [14] Those who have suffered
from acute organophosphorus poisoning frequently exhibit pin-point pupils,
profuse perspiration, lowered consciousness, and poor respiration.

Figure 4 Showing Mechanism of Action Of OP insecticides

The primary differential diagnosis is clinically indistinguishable carbamate

toxicity.[15] Organophosphates exert powerful inhibitory action over other
carboxylic ester hydrolases such as chymotrypsin, butyrlcholinesterase
(pseudocholinesterase), plasma and hepatic carboxylesterases,
paraoxonases, and other non- specific protease. Organophosphates can be
absorbed by any route including transdermal, transconjunctival,
inhalational, across the GI and GU mucosa, and through direct
injection.(Rajesh Barbadle)

10
Chemicals such as organophosphate insecticides that can inactivate AChE are
known to attach to the -CH2OH residue of the esteratic site of the enzyme by
forming a covalent bond. They are therefore often called covalent inhibitors of
AChE.[wong et. al., 2019 16]

Metabolism of OP Pesticides

Many phase I and phase II xenobiotic-metabolizing enzymes (XMEs) are

involved in the metabolism of pesticides.

The chemical reactions involved in the biotransformation of pesticides may

be either detoxications or activations.

11
Diagnosis [17]

1. Choline-esterase level

RBC ChE Level: Organophosphate poisoning is indicated by

a decrease in RBC cholinesterase levels that is greater than
50% of normal. The decline is due to pesticide phosphate
group binding. It is a more effective parameter than
plasma cholinesterase.[18] RBC cholinesterase levels are
more accurate than serum cholinesterase in diagnosing
organophosphate toxicity.

Disadvantages- [18]

Normal cholinesterase levels are based on population estimates,

and the definition of normal has a wide range. A person with a

12
"high normal" level may have symptoms from a "low normal"
activity.

Several people do not appear to have a known baseline level.

A very low level of cholinesterase may not usually correlate with

clinical disease.

Pernicious anaemia, haemoglobinopathies, anti-malarial

medication, and blood collected in oxalate tubes all cause false
depression of RBC cholinesterase levels. Elevated levels can occur
as a result of anaemia, bleeding, or treatment for megaloblastic or
pernicious anaemia.

Other Body Fluids ChE level: A decrease in plasma (serum)

cholinesterase activity greater than 50% of normal implies
organophosphate poisoning. Depressions of more than 90% are
common in severe poisonings and are frequently connected with
death. This test is not selective because plasma cholinesterase
activity is similarly reduced in liver cirrhosis, neoplasia, starvation,
septicemia from burns, and obstructive jaundice.

Blood should only be obtained in heparinized tubes for the

purpose of estimating cholinesterase levels. Alternatively, frozen
samples can be used. Plasma cholinesterase normally recovers in a
few days or weeks, whereas red blood cell cholinesterase recovers

13
in a few days to four months, depending on the degree of the
depression.

2. Urinary p-Nitrophenol test[19]

P-Nitrophenol is a metabolite of some organophosphates and is excreted in

the urine .

Procedure:

Steam distilled 10mL of urine and collects the disitillate. Add sodium
hydroxide (2pellates) and heat on a water bath for 10 minutes. Production
of yellow colour indicates the presence of P- NITROPHENOL.

3. Chromatography [20 , 21, 22]: Paper Chromatography, TLC, GC,

GC-MS, HPLC

 TLC: Mobile phase: Hexane(C6H12):Acetone(CH3COCH3) =

(8:2), OR Petroleum Ether:Methanol = 25:1

 Spraying reagent: Iodine Azaide solution

Table 1 Rf values of some common OPC

OP Compound Rf-value (Silicagel- Rf-value (Silicagel-

Ethylacetate) dichloromethane)

Malathion 0.62 0.33

14
Methyl-parathion 0.64 0.56

Chloropyriphos 0.65 0.69

Fenthion 0.64 0.62

Monocrotophos 0.06 0.00

4. Colorimetric Method:

1 mL of urine is collected, and 1 mL of NBB 45% in acetone 4-(nitrobenzyl)

pyridine is added and stirred in a vortex mixer for 30 seconds.

5. Ancillary investigations like increase blood sugar level,

leukocytosis, high hematocrit, and anion gap acidosis.

Medico-legal Importance [23]

 Accidental poisoning can occur in farmers while spraying in the fields

or opening the lids of containers.
 Suicidal poisoning is common with this Organophos-phorous
insecticide.
 Homicidal is rare as it is difficult to mask the smell Organophos-
phorous insecticide.

15
Forensic Issues [24 - 34]

Acute pesticide poisoning is a huge global problem, causing an estimated 3

million cases of severe poisoning each year and killing roughly 220,000
people. More than 90% of these reports come from underdeveloped
countries like India. Among the different pesticides, organophosphates
account for the majority of documented cases of poisoning particularly in
Northern and Central India. Suicidal pesticide exposure is almost always
done orally, while a few cases have involved self-injection.

The horrible potential for pesticides to cause mass poisoning was not
recognised in India until the Kerala food poisoning disaster in 1958. Wheat
flour and sugar were unintentionally housed in the same cabin on a ship as
parathion, which absorbed into the former's containers. Over a thousand
people were poisoned as a result of consuming the tainted substance, and
over a hundred died as a result. More recently (late 1970s), a severe
convulsive epidemic in Uttar Pradesh killed hundreds of people due to
horrible ignorance on the part of farmers who were preserving food crops
with pesticides. Another (recent) example includes the unintended spraying
of concentrated pesticides in a densely populated area, resulting in
comparable mass morbidity.

Unfortunately, history does not teach us effectively, and cases of

irresponsible storage of food grains, sugar, vegetables, and fruits, as well as
harmful pesticides, resulting in large-scale contamination are not unusual
even now. Pesticide contamination of crops is a much greater issue in India
than in many other countries. In fact, pesticide contamination of food due to
excessive crop spraying or improper storage is so common in India that
blood samples and milk samples (from lactating mothers) collected at

16
random from several people in one study revealed residual insecticide levels
in the majority of the samples.

Pesticide exposure can occur in a variety of ways. A study of offices sprayed

with diazinon, chlorpyrifos, or bendiocarb found that surface concentrations
were higher 24 or 48 hours after spraying than one hour afterwards.
Occupants should be informed about treatment times and efforts taken to
reduce exposure. While caring for patients, health care providers may be
exposed to Section 8 Hydrocarbons and Pesticides 406 cholinesterase
inhibitors. Hospitals must prepare for such events by stockpiling atropine,
disposing of poisonous waste, and installing individual ventilation systems.
Chronic liver problems have been observed in Vietnam soldiers who may
have been exposed to chlorophenoxy herbicides.

Organophosphates that operate quickly, like as tabun, sarin, soman, and VX,
have been created as "nerve gases" for chemical warfare. Terrorist activities
has resulted in human exposure. The most noteworthy case of sarin use by a
terrorist cell happened in March 1995, when sarin was released in Tokyo's
underground system. The attack is said to have affected 1,000 individuals,
with 12 persons killed.

Organophosphates can be purposefully consumed by cocaine addicts in an

effort to extend the effects of the drug by lowering cholinesterase activity.
Organophosphate poisoning symptoms have been documented in this
situation.

Several clauses have been inserted into the Insecticides Act of 1968 and the
Insecticides Rules of 1971 to reduce the occurrence of accidental poisoning

17
due to occupational exposure, irresponsible handling, and contamination of
water, milk, and food things.

Pesticides such as organophosphates and carbamates have long been

particularly popular in India for the purpose of suicide due to their ease of
access. Pesticide poisoning has traditionally been uncommon due to the
unpleasant odor/taste that most of these compounds have. However,
incidents have been observed with other chemicals as well. According to
one case report, paraquat was used to murder two children.

This thesis is basically aiming to finding the male:female ratio, age group
ratio and locality or area of case registered which will be beneficial for
further studies and updated knowledge about the OPC poisoning in that
particular area and age people and measures to be taken for the controls of
OPC poisoning in such conditions.

18
 Section-Second

Review Of
Literature

19
Organophosphate poisoning (OPP) remains a significant public health
concern in many regions worldwide, particularly in agricultural communities.
The incidence of OPP is of special relevance in Uttar Pradesh, India, because
agriculture is important to the economy and daily lives of the majority of the
population. The goal of this review of the literature is to give readers an
overview of the current knowledge and research on the trends in
organophosphate poisoning in Uttar Pradesh. It is essential to comprehend
the historical and current context of OPP in this in order to develop
successful preventative efforts and enhance healthcare outcomes. So,
before going for observational studies, we need to gain some knowledge
through literature review in a systematic way which has been done in
Organophosphate insecticides.

In 1847, P. E. THENARD [35] published a paper on phosphines. At the same

time, M. CLOEZ identified a thiophosphoric acid ester; he had suspected the
existence of sulphur derivatives of phosphorus in parallel to arsenic. In 1854,
PH. DE CLERMONT, encouraged by WURTZ, synthesised tetraethyl
pyrophosphate (later TEPP) by alkylating the silver salt of pyrophosphoric
acid using alkyl halides (Clermont technique). MOSCHNIN is claimed to have
prepared TEPP at WURTZ's laboratory earlier. Clermont, on the other hand,
did not recognise the compound's important physiological function. During
World War II, B. C. SAUNDERS and his colleagues discovered only mild
insecticidal properties until he reacted the dichloridates with potassium
cyanide, yielding the highly poisonous and miotic Tabun.

PON1 levels in adults were shown to vary up to 13-fold in 2006, and

sensitivity to diazoxon was found to vary up to 26 and 14-fold in a sample of
neonates and Latino mothers, respectively. This considerable variety in

20
enzyme levels influencing a human's sensitivity to various OPs is being
investigated further.

According to the world health organization (WHO) report 2012 [37],

worldwide, there were about 193,460 deaths due to unintentional poisoning
and 370,000 deaths due to suicidal pesticide poisoning.

In 2014 the data study of trends of pesticide poisoning of past 13 years was
conducted in for National poison Information Centre (NPIC) AIIMS, New
Delhi and this study have also highlighted the most common classes of
pesticides that causes poisoning which includes OP as one of the most
common class that was related with such incidents in household poisonings.
It is also evident for the intentional poisoning as one of the common mode
of poisoning followed by accidental poisoning.[38]

The presentation pattern of cases of acute organophosphorus (OP)

poisoning was conducted in a tertiary care hospital in South India in 2014.
The study was conducted through the hospital records of individuals with
acute OP poisoning.

Recently, The Indian Express report says lots of hue and cry occurs due to
death of farmers in Maharashtra due to inhalation of insecticides while
spraying in the fields.[39]

Epidemiology of Organophosphate Poisoning

Pesticides belonging to the class of compounds known as organophosphates

are regularly linked to poisoning incidents. The epidemiology of OPP in Uttar
Pradesh illustrates the complex interactions between agricultural practises,
pesticide use, and the availability of healthcare services. This scale was
developed by N Senanaayake et al.[1993] [40] to assess the severity of OPC

21
poisoning. In general patients suffer from respiratory failures (giyanwani et.
al.).[41] In a clinical study, Eddleston et al. [42] have categorized their
findings of respiratory failure into two categories: early onset and the late-
onset. Banday et al. [43] have reported that the mortality was higher in
patients who required ventilatory support for the duration of more than
seven days

The Poisons Information Centre is critical in providing prompt management

guidance, including the supply of essential antidotes from the recently
established National Antidote Bank, thereby saving lives. A three-year
retrospective examination of poisoning responses collected by the National
Poisons Information Centre revealed a total of 2719 calls (April 1999-March
2002). This study also attempted to cover the cases of children. [Srivastava
et. al., 2005] Scientific evidence indicates that the relationship between
environmental toxicity and psychiatry has significant public health
consequences. [Jaga et. Al., 2007]

WHO estimates more than 3 million cases of acute poisoning and 3 lakh
deaths every year globally occurs and India is among the highest in such
incidence of poisoning in the world.[b1]

Pesticides for homes and farms frequently use dichlorvos. In developing

nations, it is the most widely applied organophosphate insecticide (Binu
Kumar and Gill, 2010). Research has also shown that OPP incidence varies
throughout districts and during different seasons, indicating the necessity
for localized treatments (Tripathy et al., 2017). The high prevalence of OPP
in this area has been noted in several studies, with a spike that is particularly
noticeable during particular farming seasons when pesticide application is at
its highest (Tripathy et al., 2017; Singh and Sharma, 2000).

22
(Rahman et. al., 2021) Performed A clinic-epidemiological study of
organophosphorus poisoning observed that most occurrences of
unintentional poisoning are avoidable by keeping a watchful eye on infants
and younger children who have a propensity of tasting unknown items.

Risk Factors and Contributing Factors

The risk of OPP is greater for agricultural workers since they frequently
come into direct contact with pesticides. Organophosphate toxicity can
result in neurobehavioral effects like depression, a common psychiatric
disorder that affects humans globally. A study suggests that the association
between environmental toxicology and psychiatry has important public
health implications.(Jaga et.al. , 2007) Specific risk factors have been
discovered by studies in Uttar Pradesh, such as a lack of appropriate
protective equipment, insufficient training, and little knowledge of pesticide
safety precautions (Tripathi et al., 2018). Additionally, it has been discovered
that socioeconomic conditions, such as poverty and illiteracy, increase
sensitivity to OPP (Vachhrajani et al., 2019).

Suicide is a complicated phenomena influenced by a variety of causes that

affect both individuals and society.(Jacob, 2008)[46]Young males and
females of rural background with agriculture occupation, socio-economic
problems are the risk factors associated with poisoning cases (Shahina et.al.,
2023). For the purpose of developing focused preventative methods, it is
crucial to investigate the intricate network of risk variables.

Acute organophosphorus (OP) pesticide poisoning is common in

underdeveloped countries. Pesticides and insecticides are widely employed

23
in agriculture in this largely agricultural country, and the people has easy
access to these harmful chemical chemicals. Organophosphate poisoning
can present clinically in a variety of ways, ranging from mild symptoms such
as nausea and vomiting to severe ones such as respiratory distress and
neurological problems. Uttar Pradesh, like many other states, faces
challenges in identifying and managing OPP cases quickly. Limited access to
healthcare facilities, delays in seeking care, and a lack of medical resources
may all have a negative impact on outcomes (Saxena and Sharma, 2015).
According to study, early diagnosis and successful treatment are critical for
lowering the region's OPP-related fatality rates.

Regulating pesticides and ensuring safety standards are crucial steps in

lowering OPP in Uttar Pradesh. The state government has implemented a
number of rules and regulations to limit pesticide use and protect
agricultural workers. However, studies have revealed that monitoring,
enforcement, and implementation all have flaws (Sharma and Kumar, 2019).
A complete examination of the current legal and regulatory environment is
required to identify areas for improvement and tighten pesticide safety
standards.

Insecticide Act 1968:[44] The pesticides Act of 1968 is the principal piece of
legislation in India that governs the manufacture, import, sale,
transportation, distribution, and use of pesticides. This category includes
organophosphate insecticides. To regulate and monitor pesticides, the Act
creates the Central Insecticides Board (CIB) and the Registration Committee
(RC).

Central Insecticides Board (CIB) and Registration Committee (RC): These

are the regulatory bodies established by the Insecticides Act. They are in

24
charge of pesticide registration and re-registration, determining residue
limits, and ensuring pesticides satisfy safety and efficacy criteria.

Knowledge Gaps and Future Directions

Many regions, including Uttar Pradesh, lack comprehensive and up-to-date

epidemiological data on organophosphate poisoning, making it difficult to
assess the true burden of the problem. While some risk factors are known,
there is a need for more in-depth research into the complex interplay of
factors that lead to organophosphate poisoning, as well as the identification
of specific vulnerable populations (Karunarathne et. al.).[4] The long-term
health consequences of acute and chronic organophosphate exposure,
particularly in agricultural workers, are not fully understood (Eddleston et.
al.).[5] There is often a gap between pesticide regulations and their effective
enforcement in many countries, leading to the continued use of highly toxic
pesticides. Rapid and accurate diagnostics for organophosphate poisoning
are limited (Srivastava et al.).[6] Early diagnosis is crucial for prompt
treatment. While some treatments are effective, research is needed to
develop novel therapeutic strategies and optimize standardized treatment
protocols. In many regions, there is a lack of effective public health
education and awareness campaigns on the risks of organophosphates and
safe handling practices. The environmental impact of organophosphates,
including their effects on ecosystems and water sources, requires further
investigation (Marecek et. al.).[10] In underserved areas, access to
healthcare is often limited. Telemedicine and digital health solutions can
bridge this gap. Comprehensive psychosocial support services for individuals
affected by organophosphate poisoning, particularly in cases of self-harm,
are often lacking.

25
In order to overcome this challenge we need to Conduct a large-scale or
many small-scale epidemiological studies and surveillance systems to gather
accurate data on the incidence, distribution, and determinants of
organophosphate poisoning in various regions. Investigate the roles of
gender, socioeconomic status, occupation, education, and access to
healthcare in influencing the risk of organophosphate poisoning. Tailor
prevention efforts to high-risk populations. Conduct long-term cohort
studies to assess the impact of organophosphate exposure on neurological,
respiratory, and other health outcomes. Investigate potential links to
neurodegenerative diseases. Advocate for stronger, more effectively
enforced pesticide regulations and the removal of highly toxic pesticides
from the market. Monitor compliance and implement stricter penalties for
non-compliance (Aaron et. al.).[15] Invest in research to identify and
validate specific biomarkers for early detection, enabling healthcare
professionals to initiate treatment more rapidly. Investigate new antidotes
and treatment modalities that are more effective, readily available, and
affordable. Develop and disseminate standardized treatment guidelines.
Develop culturally sensitive and accessible public health campaigns to raise
awareness and educate both the general population and high-risk groups
about organophosphate poisoning risks. Research the ecological
consequences of pesticide use and assess the long-term effects on non-
target organisms, soil quality, and water resources. Promote the use of
telemedicine and digital health tools to provide remote guidance,
consultation, and support for healthcare providers in areas with limited
access to medical facilities. Establish and assess the impact of psychosocial
support services, including counseling and mental health resources, to
address the psychological and emotional needs of affected individuals.

26
Addressing these knowledge gaps and pursuing these future directions will
require collaboration among researchers, healthcare professionals,
governments, non-governmental organizations, and international bodies. It
is through such collaborative efforts that we can advance our understanding
of organophosphate poisoning and develop effective strategies to mitigate
its impact on public health.

27
 Section-Third

Methodology

28
Organophosphate poisoning is a significant public health concern in many
agricultural regions, including Uttar Pradesh, India. This study aims to
investigate the trends in organophosphate poisoning cases in Uttar Pradesh
over the past months to understand the epidemiology, risk factors, and
outcomes associated with this poisoning. The findings will contribute to the
development of effective preventive and management strategies.

In order to get confirmation of the type of poison present in the body of the
person, viscera or various body fluids or in some cases both are preserved
and sent to the laboratory.

Protocols followed by laboratory in order to confirm the type of pesticide

has been injected into the patient are as per the standard procedures of
state government and DFSS manual. We have followed the chemical
extractions and TLC method for the confirmation test of Organophosphates
into the extracts of the preserved sample sent by the Medical unit or legal
unit.

Sample extraction in Insecticide

We have followed the extraction by standard protocol of chemical method

for extraction which is given below. Here sample is cutted piece of viscera
which has been sent from Medical unit.

Sample(50gm) + Na2SO4  Shake well and transfer in conical flask Add

100 ml n-Hexane  Heat the sample now for 1 hour with adding Air
Condenser  Cool and Filter  Transfer filterate in separating Funnel 
Extract 3 times with 50, 25, 25 ml of acetonitrile saturated by n-Hexane 
Add 500 ml dist. H2O  Add 25 ml Saturated Na2SO4 solution  Add n-

29
Hexane again  heat so that n-Hexane get evaporated  the extrtact is
final extract

Confirmation of the presence of type of insecticide in the Sample

In the above extract add 8-9 ml Hexane and shake well into the separating
funnel, the liquid will get separated into upper and lower half. Put the liquid
of Lower Half out into the beaker and upper half liquid into the conical flask.
This conical Flask sample is ready for TLC analysis.

Thin Layer Chromatography (TLC)

Thin Layer Chromatography is a

traditional method which uses
chromatography principles for
the detection and separation of
components of the sample. This
separation of components of
shows a particular Rf-values (as discussed in introduction). This confirms tha
type of compound. There is another way which we have used in the
laboratory that is the sensitivity of the sample towards the spraying reagent.
TLC plate in case of OP compounds used are Silica gel coated with the
aluminium plate with the help of ethylacetate. Using the capillary tube we
have implemented the sample over the plate at the certain distance from
one side of the plate as per standard protocols. These may also have been
implemented during the solved case files. The plate is set for analysis into
the beaker with the solvent of Hexane:Acetone in the ratio of 8:2 during the
training. For Organophosphate Compounds after TLC has been acted with

30
the solvent into the beaker, Sprayed with Paladium Chloride (PdCl2) solution
or we may also choose various other reagents which are discussed in
introductory section. The samples with OPC show visibility or sensitivity with
the UV-Visible light after sprayed with PdCl2 solution.

Study Objectives:

The objectives of this retrospective observational study are as follows:

 Primary Objective: To determine the trends in organophosphate

poisoning cases in Uttar Pradesh over the certain period of study
during one month internship. Also to assess the seriousness of the
application of the substance studied.
 Secondary Objective: To assess the factors contributing to
organophosphate poisoning. Also to provide recommendations for
prevention and management strategies based on the study findings.

Study Design

Study Setting

The study has been conducted in Uttar Pradesh, one of the largest states in
India with a high prevalence of agricultural activities and a significant risk of
organophosphate poisoning.

Study Period:

Data has been collected retrospectively for the period of a month, from June
27, 2023 to July 26, 2023.

31
Data Sources:

Data for this study is obtained from multiple sources during the training
period in SFSL, including:

 Records from Forensic Science Laboratory, Lucknow, Uttar Pradesh.

 Poisoning control centers i.e. AIIMS.
 Mortality records from the Registrar General of India like NCRB.
 Government health department records.

Study Population:

The study will include all individuals with confirmed cases of

organophosphate poisoning in Uttar Pradesh during the study period. Both
male and Female population has been included with all the age group
patients are considered into the study during the one month training at
SFSL.

32
 Section-Fourth
Data Collection

33
This thesis is regarding retrospective observations of study of
organophosphates in Uttar Pradesh. As per the reviews of the previous
studies have been conducted from in Critical care units and the data
available on internet. This retrospective analysis was carried out in
Toxicology Division of SFSL, Lucknow, Uttar Pradesh, India. From June 2023
to July 2023, Laboratory records of solved cases were examined. Age, sex,
and locality/city region were recorded in a systemic examination, laboratory
investigations.

Variables:

The following variables will be collected from the data sources:

 Age group [From FSL]

Table 2

Age Group Number of cases

5-20 7
21-35 27
36-50 28
50+ 6

 Sex or Gender [From FSL]

Table 3

Gender Number Of Cases

Male 34
Female 24

34
  Residential City or Town [from training FSL]

Table 4

S. no. Region Number Of Cases

1 Ayodhya 4
2 Barabanki 6
3 Lucknow & Kanpur 24
4 Raebareli 2
5 Sitapur 9
6 Hardoi & Shajehanpur 10
7 Lakhimpur 13

Data Collection Methods:

Data has been extracted from file records and databases including internet
sources. Data collectors are trained in data extraction procedures.

35
 Section-Fifth

Data
Analysis
And Result

36
Data Analysis:

Statistical Methods:

Descriptive statistics is used to summarize the characteristics of

organophosphate poisoning cases. Trends over time has been
assessed.

 A total of 68 samples have been collected which shows distinct

variability distributed among age, sex and region.
 Age group [From FSL]: After interpreting the age group we found the
percentage of cases of dead people by organophosphate poisoning.
On the basis of calculation of data the pie chart has been maintained.
 Calculation of percentage of death among various age group found
after the collected data are given below.

𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑐𝑎𝑠𝑒𝑠 𝑖𝑛 𝑠𝑒𝑙𝑒𝑐𝑡𝑒𝑑 𝑣𝑎𝑟𝑖𝑎𝑏𝑙𝑒 × 𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑣𝑎𝑟𝑖𝑎𝑏𝑙𝑒𝑠

𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑜𝑓 𝑣𝑎𝑟𝑖𝑎𝑏𝑙𝑒𝑠 =
100

 By applying this formula in the above equation as we have number of

selected variables among the age group as in :
o Between 5-20 the number of selected variables are 7 out of
total of 68 variables as collected samples which shows the 10%
of population affected.
o Between the age group of 21-35 the number of selected
variables are 27 out of 68 collected samples which shows the
40% of population affected.
o Between the age group of 36-50 the number of selected
variables are 28 out of 68 collected samples which shows the
41% of population affected.

37
 o Above the age group of 50+ the number of selected variables
are 6 out of 68 collected samples which shows the 6% of
population affected.
 The above calculated percentage shows that the population between
the age of 21-50 years are highly affected

Number of cases

6 7

28 27

05 to 20 21 to 35 36 to 50 50+

Chart 1: Chart showing the Age group

 Sex or Gender [From FSL]

o Out of total population selected 34 are male population which
is 59% of total sample collected and 24 are female population
which is 41% of total population.
o Male:Female ratio is 17:12 in numbers obtained whereas 59:41
in percentage obtained.
o This shows that male population is affected comparatively.

38
 Number Of Cases

24

34

Male Female

Chart 2 Chart shows the affected male and female population

 Geographical Region of the Sample variables [from FSL]

o Cases from Ayodhya region is 6% of the total collected samples.
o Cases from Barabanki region is 9% of the total collected
samples.
o Cases from Lucknow and Kanpur Region is 35% of the total
collected samples.
o Cases from Raebareli region is 3% of the total collected
samples.
o Cases from Sitapur region is 13% of the total collected samples.
o Cases from Hardoi and Shahjehanpur region is 15% of the total
collected samples.
o Cases from Lakhimpur region is 19% of the total collected
samples.

39
 Number Of Cases
30

25

20

15

10

0
Ayodhya Barabanki Lucknow & Raebareli Sitapur Hardoi & Lakhimpur
Kanpur Shajehanpur

Number Of Cases

Ethical Considerations:

All the suspected data taken after the concern given by the FSL
personnel during the time of training that is why it’s a
retrospective observational study hence no ethical concern
required.

40
 Section-Seventh

Conclusion

41
After the detailed observational study, literature reviews and various
resources during the study, the data collected and interpreted statistically as
well as the result obtained, and as per the objective of this thesis study I can
conclude the following:

 In the case of age we can see that major age group affected is
between 21 to 35 and 36 to 50 which is highly affected. As per the
literature reviews major rural population is affected so need for the
proper education about the pesticides and their uses and measures to
be taken during the agricultural use.
 During the study of gender variation we found that male population is
affected comparatively which shows that lack of proper knowledge,
easy availability of the such product in cheap prices in the locality,
also many a times family disputes which should not be
underestimated.
 During the study or geographical distribution of variables maintained
by the sample collection in the chart discussed earlier the population
in the region of Awadh belt of Uttar Pradesh especially in the region
of Lucknow and Kanpur is affected greatly.
 Government must include the pedsticide studies and farming in the
courses of matriculation level students as compulsory especially in the
rural regions.
 Government must have to take the measures for the hospitals and
critical care units and consistently look after whether the hospital is
well equipped with the facilities of emergency cases of poisonings or
not and

42
Limitations:

This study is subject to limitations such as potential underreporting of cases,

data quality issues, and biases inherent in retrospective studies. The sample
collected is only depend upon the cases registered earlier. The sample has
been taken only those which has been taken for the analysis. The sample
taken only during the one month training in SFSL Lucknow, if the sample will
be collected from various other sources such as Emergency or critical care
units of Uttar Pradesh, that may vary the result. This could be the major
research gap in this study.

43
 Section-Eighth

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