• Foods and drink can cause people to be ill because they are

contaminated with harmful micro-organisms and/or their

toxins (poisons) - this is often called food poisoning.
poisoning

• To detect : food samples

• Two major processes:

– To detect the presence of a toxicant in foods
– To qualitate and quantitate the toxicants
• Food safety : An important step in the initial phase is
assessment to estimate the levels of the toxicant to
which the population is exposed.

☼ Method: a) dietary survey

b) market basket analysis

• chemical analysis in food toxicology involve

separating a toxicant from other chemicals and then
determining the amount
 sampling

 To obtain a portion for the estimation or

observation of attributes of the particular lot; the
sample must be representative of the lot.
 Qualitative and quantitative

 Require both an assay for detecting the poison and a

method for separating it from the rest of the chemicals in
the foods

 Separation methods:
 Gas chromatography (GC)
 High performance liquid chromatography (HPLC)
 Column & thin layer chromatography (CC, TLC)
 Distillation
 extraction
chromatography
 GC
 HPLC
 CC, TLC
 a way of separating out a mixture of chemicals, which are in
gas or liquid form, by letting them creep slowly past another
substance, which is typically a liquid or solid

 involve the interaction of a mixture between a stationary and

a mobile phase

 The key thing to remember is that chromatography is

a surface effect.
Different types of chromatographic techniques.
Gas chromatography
 used in organic chemistry for separating and analyzing compounds that
can be vaporized without decomposition.
 include testing the purity of a particular substance, or separating the
different components of a mixture
 moving phase (or  mobile phase) is a carrier gas, usually an inert gas such
as helium or an un reactive gas such as nitrogen.
 stationary phase is a microscopic layer of liquid or polymer on an inert
solid support, inside a piece of glass or metal tubing called a column.
 High performance liquid
chromatography
 utilizes a column that holds chromatographic packing material
(stationary phase), a pump that moves the mobile phase(s)
through the column and a detector that shows the retention times
of the molecules.

 Retention time varies depending on the interactions between the

stationary phase, the molecules being analyzed and the solvent(s)
used.

 one of the most powerful tools in analytical chemistry. It has the ability
to separate, identify, and quantitate the compounds that are present in
any sample that can be dissolved in a liquid.
 Column chromatography
 to purify individual chemical compounds from mixtures of
compounds.
 used for preparative applications on scales from micrograms upto
kilograms, and for cleaning the analyte of the matrix
interferences, for use with more sophisticated techniques
 Thin layer chromatography

 used in separation of mixtures. TLC is performed on a sheet of

glass, plastic, or aluminum foil, which is coated with a thin layer of
adsorbent material, usually silica gel, aluminium oxide, or
cellulose.

 This layer of adsorbent is known as the stationary phase.

 After the sample has been applied on the plate, a solvent or

solvent mixture (known as the mobile phase) is drawn up the plate
via capillary action. Because different analytes ascend the TLC
plate at different rates, separation is achieved.
Biological Determination of
Toxicants
 Genetic toxicity
 Acute toxicity
 Bioassay
 Metabolism
 Subchronic toxicity
 Teratogenesis
 Chronic toxicity
 Definition:

•refers to the ability of substances or physical agents to

damage the DNA and/or chromosomes of cells. Such
damage can lead to mutations that increase the
likelihood of certain diseases, such as cancer and birth
defects.

•determined using a wide range of test species

including whole animals and plants (e.g., rodents,
insects, and corn), microorganisms, and mammalian
cells.
 The most common gene mutation tests involve:
Acute toxicity

Definition:

 the toxicity produced by pharmaceutical when it is

administered in one or more doses during a period
not exceeding 24 hours.
 Acute toxicity tests are generally the first tests
conducted. They provide data on the relative
toxicity likely to arise from a single or brief exposure.
Standardized tests are available for oral, dermal,
and inhalation exposures.
Purpose
 to identify the dose of the
pharmaceutical which causes
major adverse effects, to help
set doses for further preclinical
studies and to predict the
effects of overdose in humans.
Basic parameters of these tests
are:
 Bioassay
Definition:

 the use of a living organism to test for the presence

of a compound or to determine the amount of the
compound that is present in a sample.
 a procedure for the determination of the
concentration of a particular constituent of a
mixture.
 a measurement of the effects of a substance on
living organisms.
 Types of Bioassays

 [1] Quantal Assays [ Direct endpoint ]

 Elicits an ‘All or None’ response in different animals
 Eg.
 Digitalis induced cardiac arrest in guinea pigs
 hypoglycemic convulsions in mice.
 Digitalis induced head drop in rabbits
 Calculation of LD50 in mice or rats

 [2] Graded Response Assays [mostly on tissues]

 Graded responses to varying doses
 Unknown dose response measured on same tissue
Examples of Biological Assay:

 Limulus Test
a sensitive method for detection of bacterial
endotoxins and endotoxin.

 Endpoint Determination
an establishment of the level of a quantifiable
effect indicative of a biologic process. The
evaluation is frequently to detect the degree of
toxic or therapeutic effect.
Metabolism
 First, Santorio in 1614 in his book Ars de statica
medicina( vital force which later known as metabolism)
 Metabolic studies can be conducted after mutagenicity
test.
 Objective:
 To gain understanding of the:
 Absorption,
 Biotransformation,
 Disposition (storage),
 Elimination characteristics of an ingested substance after single
and repeated doses.
Metabolism

 Knowledge of the metabolism and

pharmacokinetics of a substance is essential
for establishing the relevance of results from
animal testing to projecting likely hazards in
humans.
Subchronic toxicity

 Subchronic Toxicity - ISO 10993-11

Studies that continue for 90 days or for up to
10% of a test subjects life span are considered
subchronic. Studies that continue for longer
than 10% of a test subjects life span are
considered chronic.
Subchronic toxicity

 Objective
 Determine the possible cumulative effects on
tissue or metabolic system
 To evaluate the safety of food components.
 Determine if test substance toxic or not.
 Performed for several months duration.
Subchronic toxicity

 Procedure :
 Inspection of physical appearance and behavior
 Body weight
 Food consumption
 Characteristics of excreta
 Blood, urine, hepatic, body temperature renal and
more.
Teratogenesis
 Birth defects are known to occur in 3-5% of all newborns.
 They are the leading cause of infant mortality in the United States,
accounting for more than 20% of all infant deaths.
 7% to 10% of all children will require extensive medical care to diagnose or
treat a birth defect.
 And although significant progress has been made in identifying the etiology
of some birth defects, approximately 65% have no known or identifiable
cause.
 It was previously believed that the mammalian embryo developed in the
impervious uterus of the mother, protected from all extrinsic factors.
 However, after the thalidomide disaster of the 1960s, it became apparent
and more accepted that the developing embryo could be highly vulnerable
to certain environmental agents that have negligible or non-toxic effects to
adult individuals.
What is teratogenesis ?

 Teratology is the study of abnormalities of

physiological development. It is often
thought of as the study of birth defects, but it
is much broader than that, taking in other
developmental stages, such as puberty; and
other life forms, such as plants
 
Wilson's 6
principles
 These principles of teratology were put forth
by Jim Wilson in 1959 and in his monograph
Environment and Birth Defects. These
principles guide the study and understanding
of teratogenic agents and their effects on
developing organisms:
Teratogenic agents
 A wide range of different chemicals and environmental factors are
suspected or are known to be teratogenic in humans and in animals. A
selected few include:
 Drugs and medications: tobacco, caffeine, drinking alcohol (ethanol),
isotretinoin (13-cis-retinoic acid, Roaccutane).
 Environmental chemicals: polychlorinated biphenyls (PCBs),
polychlorinated dibenzodioxins a.k.a dioxin, polychlorinated
dibenzofurans (PCDFs), organic mercury, ethidium bromide.
 Ionizing radiation: background radiation, diagnostic x-rays, radiation
therapy.
 Infections: cytomegalovirus, herpes virus, parvovirus B19, rubella virus
(German measles), syphilis, toxoplasmosis, Venezuelan equine
encephalitis virus.
 Metabolic imbalance: alcoholism, endemic cretinism, diabetes, folic
acid deficiency, iodine deficiency, hyperthermia.
Chronic toxicity
 chronic toxicity: the adverse effects occurring after the administration of
a test sample for a major part of the lifespan (usually 6-12 months).
 Chronic toxicity represents cumulative damage to specific organ systems
and takes many months or years to become a recognizable clinical
disease.
 With repeated exposures or long-term continual exposure, the damage
from these subclinical exposures slowly builds-up (cumulative damage)
until the damage exceeds the threshold for chronic toxicity. Ultimately,
the damage becomes so severe that the organ can no longer function
normally and a variety of chronic toxic effects may result.
 Examples of chronic toxic affects are:
 cirrhosis in alcoholics who have ingested ethanol for several years
 chronic kidney disease in workmen with several years exposure to lead
 chronic bronchitis in long-term cigarette smokers
 pulmonary fibrosis in coal miners (black lung disease)
 Objective:
 To assess toxicity resulting from long-term, relatively low-level
exposure, which would not be evident in subchronic testing.

 Test designed so that each treated and control group will include
sufficient numbers of animals of both sexes of the chosen species
and strain to have an adequate number of survivors.
 The chronic toxicity test provides the final piece of biological
information on whether to accept or reject a substance suggested
for food use.
 If no carcinogenic effects are found, this information will be used in
the overall risk assessment of a substance