1. Drug abuse and toxicology screening
Dr. Mohamed AL-Tufail
Head of toxicology laboratory, KFSHRC

2. 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

3. ALCOHOL

4. Equipment: Headspace GC-MS
Alcohol Analysis
Equipment: Headspace GC-MS
BLOOD ALCOHOL
URINE ALCOHOL

5. 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.

6. 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.

7. (cocaine & methamphetamine, dextroamphetamine, methylphenidate)
Psychostimulants
(cocaine & methamphetamine, dextroamphetamine, methylphenidate)

8. (cocaine & methamphetamine, dextroamphetamine, methylphenidate)
Basic Pharmacology of Psychostimulants
(cocaine & methamphetamine, dextroamphetamine, methylphenidate)
Indirect acting sympathomimetics: block the reuptake of the neurotransmitters: dopamine, norepinephrine and serotonin

9. - Chronic: psychotic delusions and paranoia.
Basic Pharmacology of Psychostimulants (con’t)
- Acute: arousal, euphoria, agitation, restlessness, insomnia, anorexia, tachycardia, hyperthermia, seizures.
- Chronic: psychotic delusions and paranoia.

10. Marijuana and hashish

11. 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

12. 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.

13. Role of Opioids

14. 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

15. Cocaine (benzoyl ecgonine metabolite) Codeine Ethyl alcohol, ethanol
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
therapeutic dose: 3 days
chronic use: 4-6 weeks or longer
Cocaine (benzoyl ecgonine metabolite)
Up to 4 days
Codeine
Analgesic / Opiate
2 days
Ethyl alcohol, ethanol
urine: 2 to 12 hours
serum/plasma: 1 to 12 hours

16. Marijuana, Cannabinoids
DRUG
CLASS
DETECTION TIME IN URINE
Heroin
Analgesic / Opiate
2 days
Marijuana, Cannabinoids
Hallucinogen
Single use: 2 to 7 days
Prolonged, chronic use: 1 to 2 months or longer
Methadone
3 days
Methamphetamine
Stimulant
Up to 2 days
Methaqualone
depressants / sedatives / hypnotics
Up to 14 days
MDMA(methylenedioxy-
methamphetamine)
Morphine
Phencyclidine
8-14 days, but up to 30 days in chronic users
Propoxyphene
6 hours to 2 days

17. 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.

18. 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.

19. Toxicology screening and drug abuse analysis using GC-MS and LC-MS

20. Method Of Analysis Extraction Clean up Concentration
Injection on GC-MS or LC-MS
Reporting

21. 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

22. Strongly acidic (low pH) Weakly acidic (pH 5 to approaching 7)
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)

23. 1) Blood & Urine
- single basic or acidic extraction
2) Urine 3 ways extraction
- acid
- weakly basic & neutral
- basic

24. Basic Extraction – GC/MS Screen Blood + Base (NH4OH) + Toluene 
BACK EXTRACTION (clean up)
Toluene + Acid (H2SO4)
Aqueous fraction + Base (NaOH) + Toluene
GC/MS

25. CHROMATOGRAPHY
chromatography is a separation process that is achieved by distribution of the substances between a mobile phase and a stationary phase
Amobile  A stationary

26. Mass Spectrometry Powerful detector can be teamed up with GC or HPLC
GC/MS LC/MS
Requires very low pressure (10-5 Torr)

27. Cocaine and its Metabolites sampling

28. Acid and base properties of cocaine and its metabolites

29. Drugs of abuse detected in human hair, methods of detection

30. Drugs of abuse detected in human hair, methods of detection

32. LC-MS/MS

34. Instruments
LC-MSMS
QTOF

36. LCMSMS (TSQ VANTAGE)

37. LCMSMS TSQ (QUANTUM ACCESS MAX)

38. Using GCMS instrument in Toxicology screening

39. Examples of toxicology screening

41. Sibutramin

42. Sample Number M:34-141 ( Capsules )

43. High Quantity of Procymidone ( Pesticide )
Containing
High Quantity of Procymidone ( Pesticide )
Procymidone concentration is 2700 ppb
Maximum residue limit is 100 ppb .
Procymidone

44. Sample Number M:34-153 ( Herbal Powder )

45. Contains
Acetaminophen
2-Pyridinamine
Methyl Salicylate

46. Sample Number M:35-214 ( Herbal Powder )

48. containing Synthetic drug Tadalafil in Concentration of 1.8 mg/g
Royal Honey
containing Synthetic drug Tadalafil in Concentration of 1.8 mg/g
Tadalafil is sexual stimulant ( Commericially Cialis )

49. Containing Synthetic drug Tadalafil
Royal Honey
Containing Synthetic drug Tadalafil
Tadalafil is sexual stimulant ( Commericially Cialis )

50. Sample Number M:34-152 ( Sliming )

51. Sibutramine

52. Sample Number M:
Herbal Powder

53. Contaminated with Aerobic plate count :
2.0x105 CFU/mL Limit CFU/mL
Yeast and Moulds :
4.0x103 CFU/mL Limit is CFU/mL
Enterobacteriaceae :
1.0x103 CFU/mL Limit is CFU/mL
Also contains high cocentration of Arsenic ( 411 ppb )
And high concentration of Lead ( 6472 ppb )

54. THANK YOU