drug abuse and toxicology screening dr. mohamed al-tufail head of toxicology laboratory, kfshrc 1
TRANSCRIPT
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Drug abuse and toxicology screening
Dr. Mohamed AL-TufailHead of toxicology laboratory, KFSHRC
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Systematic toxicological analysis (STA) is a major part of the examination in clinical toxicology. STA is aimed at detecting and identifying all substances of toxicological relevance (i.e. drugs, drugs of abuse, poisons and/or their metabolites) in biological material. Particularly, gas chromatography–mass spectrometry (GC/MS) is a routinely applied screening and confirmation tool in STA. There is clear trend, however, to complement existing GC/MS procedures with liquid chromatography-mass spectrometry (LC/MS) assays. One competent LC/MS-based approach for untargeted screening analysis makes use of MS/MS under data-dependent acquisition control (DDA) to identify compounds by subsequent library search
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ALCOHOL
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Alcohol Analysis
Equipment: Headspace GC-MS
BLOOD ALCOHOL URINE ALCOHOL
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Basic Pharmacology of Alcohol (ethyl alcohol)
Non-potent compound: 80 mg/dL (0.08%)
Nonspecific compound: (1) Potentiates the inhibitory effects of the transmitter GABA by altering the conformation of the heteropentameric GABAA receptor and increases Chloride ion entry into neurons.
(2) Inhibits the activation of NMDA-type glutamate receptors reducing Sodium and Calcium ion entry into neurons.
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Basic Pharmacology of Alcohol (con’t)
Acute: produces dose-dependent intoxication, loss of behavioral inhibition, sedation, impaired judgment, slurred speech, ataxia. At higher doses: loss of consciousness, anesthesia, coma, respiratory depression, cardiovascular depression.
Chronic: hepatitis and cirrhosis, gastrointestinal bleeding, hypertension, thiamine deficiency.
Teratogenicity: fetal alcohol spectrum disorders.
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Psychostimulants(cocaine & methamphetamine,
dextroamphetamine, methylphenidate)
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Basic Pharmacology of Psychostimulants
(cocaine & methamphetamine, dextroamphetamine, methylphenidate)
Indirect acting sympathomimetics: block the reuptake of the neurotransmitters: dopamine, norepinephrine and serotonin
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Basic Pharmacology of Psychostimulants (con’t)
- Acute: arousal, euphoria, agitation, restlessness, insomnia, anorexia, tachycardia, hyperthermia, seizures.
- Chronic: psychotic delusions and paranoia.
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Marijuana and hashish
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Marijuana Tetrahydrocannabinol (THC) is the Active alkaloid from the cannabis plant
Currently the most commonly used illegal drug Binds and activates abundant G-protein coupled receptors in brain (CB1 and CB2); reduces neuronal excitability by:
- increasing K+ conductance and - decreasing Ca+ + conductance
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Basic Pharmacology of Opiates (heroin, morphine, oxycodone)
Mu, Kappa, and Delta types opioid receptors Mu receptor activation induces euphoria, Kappa
receptor activation produces dysphoria Enkephalins, Endorphins and Dynorphins are
the endogenous ligands that are released during stress to induce analgesia, immobility (sedation), euphoria or dysphoria.
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Role of Opioids
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Excretion Drugs are eliminated from the body either unchanged as the
parent drug or as metabolites (a changed form of the drug). Organs that excrete drugs eliminate polar compounds (water
soluble) more readily than components with high lipid (fat) solubility. The exception to this premise is the lungs.
Lipid soluble drugs are not readily eliminated until they are metabolized to more polar compounds.
Possible sources of excretion include: Breath Urine Saliva Perspiration Feces Milk Bile Hair
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DRUG CLASS DETECTION TIME IN URINE
Amphetamine Stimulant Up to 2 days
Barbiturates depressants / sedatives / hypnotics
short-acting: 2 days
long-acting: 1-3 weeks
(based on half-life)
Benzodiazepines depressants / sedatives / hypnotics
therapeutic dose: 3 days
chronic use: 4-6 weeks or longer
Cocaine (benzoyl ecgonine metabolite)
Stimulant Up to 4 days
Codeine Analgesic / Opiate 2 days
Ethyl alcohol, ethanol depressants / sedatives / hypnotics
urine: 2 to 12 hours
serum/plasma: 1 to 12 hours
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Heroin Analgesic / Opiate
2 days
Marijuana, Cannabinoids Hallucinogen Single use: 2 to 7 days
Prolonged, chronic use: 1 to 2 months or longer
Methadone Analgesic / Opiate
3 days
Methamphetamine Stimulant Up to 2 days
Methaqualone depressants / sedatives / hypnotics
Up to 14 days
MDMA(methylenedioxy- Stimulant Up to 2 days
methamphetamine)
Morphine Analgesic / Opiate
2 days
Phencyclidine Hallucinogen 8-14 days, but up to 30 days in chronic users
Propoxyphene Analgesic / Opiate
6 hours to 2 days
DRUG CLASS DETECTION TIME IN URINE
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Hair Test for Cocaine
It may have a positive result within 8 hours, or at times, only after 7 to days after cocaine use. The substance can remain on the hair for months, even until the hair is trimmed. Cannot be done by a single hair, the procedure requires a samples of hair around 5 inches long.
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Elimination of Alcohol
The liver is responsible for the elimination - through metabolism - of 95% of ingested alcohol from the body. The remainder of the alcohol is eliminated through excretion of alcohol in breath, urine, sweat, feces, milk and saliva. The body uses several different metabolic pathways in its oxidation of alcohol to acetaldehyde to acetic acid to carbon dioxide and water.
Healthy people metabolize alcohol at a fairly consistent rate. As a rule of thumb, a person will eliminate 15 ml of alcohol per hour. Several factors influence this rate. The rate of elimination tends to be higher when the blood alcohol concentration in the body is very high. Also chronic alcoholics may (depending on liver health) metabolize alcohol at a significantly higher rate than the average. Finally, the body's ability to metabolize alcohol quickly tend to diminish with age.
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Toxicology screening and drug abuse analysis using GC-MS and LC-MS
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Method Of Analysis
Extraction Clean up Concentration Injection on GC-MS or LC-MS Reporting
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The extraction depends on the pKa of each component
So we have two Methods of extraction Acid and base extraction
Analysis for toxins and drug abuse in urine or blood or hair
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Theoretically can separate drugs into 5 categories
◦Strongly acidic (low pH)◦Weakly acidic (pH 5 to approaching 7)◦Neutrals (pH around 7)◦Weakly basic (pH 8-9)◦Strongly basic (pH > 9)
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1) Blood & Urine- single basic or acidic extraction
2) Urine 3 ways extraction- acid
- weakly basic & neutral - basic
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Basic Extraction – GC/MS Screen
Blood + Base (NH4OH) + Toluene
BACK EXTRACTION (clean up)Toluene + Acid (H2SO4)
Aqueous fraction + Base (NaOH) + Toluene
GC/MS
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CHROMATOGRAPHYchromatography is a separation process that is
achieved by distribution of the substances between a mobile phase and a stationary phase
Amobile A stationary
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Mass Spectrometry
Powerful detector
can be teamed up with GC or HPLCGC/MS LC/MS
Requires very low pressure (10-5 Torr)
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Cocaine and its Metabolites sampling
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Acid and base properties of cocaine and its metabolites
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Drugs of abuse detected in human hair, methods of detection
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Drugs of abuse detected in human hair, methods of detection
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LC-MS/MS
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Instruments
LC-MSMS QTOF
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LCMSMS (TSQ VANTAGE)
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LCMSMS TSQ (QUANTUM ACCESS MAX)
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Using GCMS instrument in Toxicology screening
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Examples of toxicology screening
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Sibutramin
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Sample Number M:34-141 ( Capsules )
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Containing
High Quantity of Procymidone ( Pesticide )
Procymidone concentration is 2700 ppbMaximum residue limit is 100 ppb .
Procymidone
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Sample Number M:34-153 ( Herbal Powder )
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Contains
2-Pyridinamine
Methyl Salicylate
Acetaminophen
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Sample Number M:35-214 ( Herbal Powder )
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Royal Honeycontaining Synthetic drug Tadalafil in Concentration of 1.8 mg/g
Tadalafil is sexual stimulant ( Commericially Cialis )
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Royal HoneyContaining Synthetic drug Tadalafil
Tadalafil is sexual stimulant ( Commericially Cialis )
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Sample Number M:34-152 ( Sliming )
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Sibutramine
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Sample Number M: 34-125-1Herbal Powder
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Contaminated with
Aerobic plate count :
2.0x105 CFU/mL - Limit 105 CFU/mL
Yeast and Moulds : 4.0x103 CFU/mL - Limit is 103 CFU/mL
Enterobacteriaceae : 1.0x103 CFU/mL - Limit is 103 CFU/mL
Also contains high cocentration of Arsenic ( 411 ppb )
And high concentration of Lead ( 6472 ppb )
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THANK YOU