Analytical Technique for Broad-Spectrum Drug Detection
High-performance liquid chromatography (HPLC)
Simple, inexpensive and rapid spot tests
Immunoassays - SCREENING
Chromatographic and/or mass spectrometric techniques
a.Thin-layer chromatography (TLC)
CARBON MONOXIDE
colorless, odorless, tasteless gas that is the
product of incomplete combustion of
carbonaceous material.
Not only decreases the oxygen content of
blood.
Also decreases oxygen availability
to tissue, thereby producing a greater degree
of tissue hypoxia
Common exogenous sources of carbon
monoxide:
a.Cigarette smoke
b.Gasoline engines
c.Improperly ventilated heating units
Factors other than carboxy-hemoglobin
concentration that contribute to the toxicity:
a.Length of exposure
b.Metabolic activity
c.Underlying disease, especially cardiac or
cerebrovascular disease
Effects:
Delayed development of neuropsychiatric
sequelae which may include: Personality
changes, Motor disturbances, Memory
impairment
Treatment:
Half-life of carboxyhemoglobin is 5
to 6 hours when the patient breathes room
air
1.5 hours when the patient
breathes 100% oxygen.
Analytical Method
Gas chromatographic methods
- reference procedures, accurate
and precise even for every low
concentrations of carbon monoxide.
Spectrophotometric methods are
rapid, convenient, accurate and precise
except at very low concentrations of
carboxyhemoglobin (less than 2% to 3%).
Reference Values
Carboxyhemoglobin in:
Rural smokers are about 0.5%
Urban nonsmokers 1% to 2%
For smokers 5% to 6%
Values may be increased by about 3% in
hemolytic anemia
b.
c.Gas chromatography (GC)
d.Gas chromatography-mass spectrometry (GC-MS)
e.Liquid chromatography-mass spectrometry (LC-MS)
AGENTS THAT CAUSE CELLULAR HYPOXIA
CYANIDE
colorless gas with the odor of almond
detectable by only about 50% of the
population.
The ionized state is cyanide (CN-)
Patients exposed to toxic concentrations of
cyanide exhibit rapid onset of symptoms
typical of cellular hypoxia:
Flushing
Headache
Tachypnea
Dizziness
Respiratory depression –which
progress rapidly to:
a.Coma
b.Seizure
c.Complete heart block
d.Death if the dose is sufficiently large
Treatment:
Rapid identification of CN-as the intoxicant
followed by administration of sodium nirite
to cause formation of methemoglobin, which
avidly binds and clears CN-, and thiosulfateto
enhance clearance via metabolism.
Analytical Method
Spectrophotometric methods
Ion-selective electrode
Isotope dilution GM-MS analysis
Reference Values
Normal CN-concentration: less
than 0.2 µg/ml of whole blood.
The patients likely become
comatose when blood CN-concentration is
greater than 2 µg/ml, and concentration
greater than 5 µg/ml are lethal.
METHEMOGLOBIN-FORMING AGENTS
Heme iron is in ferric state (Fe3+),
methemoglobin is formed, and this form of
hemoglobin does not bind oxygen.
Maintain hemoglobin iron in the reduced
state is nicotinamide-adenine dinucleotide
(NADH)-methemoglobin reductase
(diaphorase I)
•Congenital methemoglobinemia - deficiency
of NADH-methemoglobin reductase.
Toxic effects are a consequence of hypoxia
associated:
Diminished O2content of the
blood
Decreased O2dissociation from
hemoglobin species in which some, but all,
subunits contain heme iron in the ferric state.
The PO2 is normal in these patients and
therefore so is the hemoglobin oxygen
saturation.
Thus, a normal PO2 in a cyanotic patient is a
significant indication for possible presence of
methemoglobinemia.
Specific therapy: Involves the administration
of methylene blue to hasten the conversion
of methemoglobin to hemoglobin.
Analytical Method
Measured by automated
multiwavelength spectrometers
Specimens should be kept on ice
or refrigerated but not frozen
Freezing results in an increase in
methemoglobin concentration
Methylene blue and
sulfhemoglobin causes spectral interference
in the measurement of methemoglobin with
some CO-oximeteres.
Reference Values
Normal concentration: less than 1.5% of total
hemoglobin.
Up to 20% cause CYANOSIS
20% to 50% may cause:
a.Dyspnea
b.Exerciseintolerance
c.Fatigue
d.Weakness
e.syncope
50% to 70%:
a.Dysrhythmias
b.Seizures
c.Metabolic acidosis
d.coma
Greater than 70% may be lethal

ETHANOL (CH3-CH2-OH)
Ethyl alcohol; grain alcohol
The principal pharmacological action of
ethanol is depression of the central nervous
system (CNS).
The CNS effects vary, depending on the
blood ethanol concentrations, from:
Euphoria and decreased
inhibitions–less than or equal to 50 mg/dl
Increased disorientation and loss
of voluntary muscle control resulting in
irregular movements-100 to 300 mg/dl
Coma and death–greater than 400
mg/dl
A Teratogen, and alcohol consumption
during pregnancy has been known to result
in a baby being born with Fetal Alcohol
Spectrum Disorders (FASD).
These effects may include:
physical, mental, behavioral,
and/or learning disabilities with possible
lifelong implications.
Other alcohol-related conditions include
Alcohol-Related Neurodevelopmental
Disorders (ARND) and Alcohol-Related Birth
Defects (ARBD)
FASD, ARND, and ARBD affects
newborns every year than Down syndrome,
cystic fibrosis, spinabifida and sudden infant
death syndrome combined.
FASD, ARND and ARBD are 100%
preventable when a woman completely
abstains from alcohol during her pregnancy.
Metabolized principally by liver
alcohol dehydrogenase (ADH) to
acetaldehyde, which is subsequently
oxidized to acetic acid by aldehyde
dehydrogenase.
After entering the circulation,
ethanol is metabolized by the liver. The
enzyme, alcohol dehydrogenase, oxidizes the
ethanol to form acetaldehyde.
Acetaldehyde formed is then excreted or
converted to acetate and in turn acetate is
converted to acetyl CoA which leads the two-
carbon molecule into the TCA cycle.
Maximum rate of ethanol
conversion in the normal adult is approx. 7 g
of ethanol per hour. This load is equivalent
to approx. 6 –10 ml of ethanol (roughly one
beer, one glass of wine, or one shot of
whiskey)
The major route of ethanol
disposal is by way of metabolism:
Approx. 90 –98 % of the ethanol
consumed is converted by the LIVER.
Remainder is excreted primarily
by the kidney, with rate of elimination in
urine being approx. 1/2 the rate of
absorption by the stomach.
Small fraction is lost through
perspiration and respiration, with lung
ALCOHOL OF TOXICOLOGICAL INTEREST
METHANOL (CH3 OH)
wood alcohol
is used as: Solvent in commercial products
(major component of a number of paints and
varnishes, also found in paint removers)
constituents of antifreeze and
window cleaning fluids, components of
canned fuel
The CNS effects of methanol are substantially
less severe than those of ethanol.
Methanol is oxidized by the liver ADH (at
about one tenth the rate of ethanol) to
formaldehyde.
Formaldehyde in turn is rapidly oxidized by
aldehyde dehydrogenase to formic acid, which
may cause serious acidosis and optic
neuropathy, resulting in blindness or death.
Serum formate concentrations correlate
better with the degree of acidosis and the
severity of CNS and ocular toxicity than do
serum methanol concentration.
Treatments for methanol intoxication:the
administration of ethanol or preferably
fomepizoleto inhibit the metabolism of
ethanol
sodium bicarbonate therapy to help
alleviate the metabolic acidosis
folateadministration to enhance
folate-mediated metabolism of formate, and
the use of hemodialysisto enhance
clearance of methanol and formate
ISOPROPANOL (CH3-CHOH-CH3)
Readily available to the general population
as a 70% aqueous solution for use as rubbing
alcohol.
It has about twice the CNS depressant action
of ethanol but is not as toxic as methanol.
Isopropanol has a short half life of 1 to 6
hours, as it is rapidly metabolized by ADH to
acetone, which is eliminated much more
slowly (17 to 27 hours), primarily in alveolar
air and urine.
Acetone has CNS depressant activity similar
to that of ethanol, and because of its longer
half-life, it prolongs the apparent CNS effects
of isopropanol.
Severe isopropanol intoxication,
like that of ethanol, has been known to
result in coma or death.
Isopropanol and its metabolite, acetone,
may be determined by headspace gas
chromatography or by nuclear magnetic
resonance (NMR) spectroscopy.
Ethylene glycol (1,2-ethanediol)
is a common component of
hydraulic fluid and antifreeze.
The immediate effects of ethylene
glycol ingestion are similar to those of
ethanol.
However, metabolism by hepatic
ADH and ALDH results in the formation of
several toxic species, including oxalic acid
and glycolic acid, which results in severe
metabolic acidosis.
This is complicated by the rapid
formation and deposition of calcium oxalate
crystals in renal tubules.
Calcium oxalate crystal formation
may result in renal tubular damage.
disposing of approx. 0.5 –1.5% of the total
ethanol load
Detection limit: 12 hours
Fatal dose: 300 –400 mLpure
alcohol consumed in less than 1 hour
Toxic level: 250 –400 mg/dl
Headspace gas chromatographic analysis –is
the method of choice for the measurement of
methanol.
fatal dose: 60 –250 mL