CC2 LAB - 8: DRUG TESTING

1. Enzymatic

2. Capillary Electrophoresis

3. Homogenous Immunoassay

4. Chromatographic Methods

Methods for Detection of Subtances of Abuse (4):

Capillary Electrophoresis (CE)

Based on different physicochemical principles of separation

Enzyme Mediated Immunologic Technique

Uses enzyme labeled drug that competes with the drug in the sample

Enzymatic

Solely dependent on presence of particular compounds

Saliva Alcohol Breath Test

Example Test for Enzymatic Method

Saliva Alcohol breath test

Alcohol is measured by assuming the possble BAC based on the saliva specimen

Capillary Electrophoresis

Diffentiate selectivity of analytes based on different physiocochemical principles of separation

Capillary Electrophoresis

-Variation of TLC

-Same as HPLC

Homogenous immunoassay

Solution does not need to undergo separation

Enzyme Multiplied Immunologic Technique (EMIT)

An example of Homogenous Immunoassay

Enzyme Multiplied Immunologic Technique

Uses enzyme labeled drug that competes with the drug in the sample

1. Thin-Layer Chromatography (TLC)

2. Liquid Chromatography-Mass Spectropscopy (LC-MS)

3. High Performance Liquid Chromatography (HPLC)

4. Gas Chromatography (GS)

Examples of Chromatographic Method (4):

1. Serum

2. Urine

3. Gastric Fluid

Sample used for TLC (3):

PH 4.5 (acidic)

Barbiturates pH level:

PH 9.0 (alkaline)

Opiaets pH level:

Thin-Layer Chromatography (TLC)

Extraction of the specific drug will be separated would be pH-dependent

Liquid Chromatography-Mass Spectropscopy (LC-MS)

For detection of poisons in acute or chronic intoxication, therapeutic drug identificaiton and quantitation, pharmacokinetics, drug metabolism studies

1. Detection of poisons in acute or chronic intoxication,

2. Therapeutic drug identification and quantitation,

3. Pharmacokinetics,

4. Drug metabolism studies

Liquid Chromatography-Mass Spectropscopy (LC-MS) is used for detection of (4):

Liquid Chromatography-Mass Spectropscopy (LC-MS)

For confirming positive test results from a screening assay for non-volatile compounds, but not the gold standard for drug abuse

High Performance Liquid Chromatography (HPLC)

Alternative to GC-MS

High Performance Liquid Chromatography (HPLC)

Allows

1. Quantitative measurement of drugs

2. Separation of same drug

Tricyclic antidepressants and its metabolites

High Performance Liquid Chromatography (HPLC) is for ___________

Gas Liquid Chromatography (GLC)

Legally accepted method for ethanol testing

Gas Chromatography with Infrared Spectroscopy

Used for detection of amphetamines

Gas Chromatography-Mass Spectroscopy (GC-MS)

the gold standard for confirmation of screening methods such as TLC and EMIT

Gas Chromatography-Mass Spectroscopy (GC-MS)

Allows for detection of low levels of drugs

Gas Chromatography-Mass Spectroscopy (GC-MS)

Very sensitive and specific for different drugs of abuse as well as its metabolites

1. Sedatives

2. Hypnotics

3. Anti-convulsants

4. CNS depressants

SHAC!

Barbiturates are (4):

400mg/day 2-3 months

Barbiturates taken at _____ produce significant dependence

Death

Abstinence/withdrawal of Barbiturates leads to:

4-7 days

Barbiturates are unaltered in the urine and could be detected for ______

1. Sedative for surgical

2. Medical procedures

3. Treatment for anxiety and sleep disorders

SEMET

Benzodiazepines are clinically used as (3):

1. Trouble in sleeping

2. GI upset

3. Feeling unwell

4. Loss of appetite

5. Sweating

6. Trembling

7. Weakness

8. Anxiety

9. Changes in perception

Benzodiazepines withdrawal symptoms (9):

Less than 1% is excreted unaltered in the urine for 3-7 days

Benzodiazepines, less than ____ is excreted unaltered in the urine for ______

1. Nasal inhalation

2. Intravenous injection

3. Free base smoking

Cocaine is self-administered through (3):

0.5-1.5 hours

Cocaine biologic half-life

Cocaine

It is a potent CNS stimulant and a local anesthetic

1. Restlless

2. Extreme stress

3. Tremors

4. Oversensitivity

5. Spasms

RETOS

Cocaine brings (5):

1. Fever

2. Unresponsiveness

3. Difficulty in breathing

4. Unconsciousness

FUDU

Large amounts of Cocaine brings (4):

24-48 hrs after exposure

Benzoylecgonine is detected for ____ after exposure

5-8 hrs exceed 300ng/ml

Biologic half-life of Benzoylecgonine

Methylenedioxymethamphetamine (MDMA)

Designer drug

Methylenedioxymethamphetamine

Synthesized in 1914 by a German drug company

Methylenedioxymethamphetamine

Treatment of Obesity

1. Increased sensitivity of light

2. Difficulty in focusing

3. Clenching of jaws

4. Blurred visions

INCB

Methylenedioxymethamphetamine produces perceptual changes such as (4):

Methylenedioxymethamphetamine

Not stimulant but effect is same as amphetamine drug

1. Blood rate

2. Blood pressure

3. Heart rate

Methylenedioxymethamphetamine increases (3) of the individual:

1. Serotonin

2. Dopamine

Methylenedioxymethamphetamine mechanism of action is due to release of neurotransmittter (2):

Methadone

Narcottc pain reliever for medium to severe pain

Between 12-48 hrs

Methadone lasts between _____

1. Heroin

2. Opiate dependence

3. Vicodin

4. Perocet

5. Morphine

Methadone is used in the treatment of (5):

Methadone

Currently an acceptable method for

1. Detoxification of patient

2. Undergoing therapy

Morphine

Major metabolic product of codeine and heroine

1. Codeine

2. Heroin

Morphine is the major metabolic product of (2):

Morphine

Opiod analgesics which control pain by depressing the CNS

Morphin

Produces higher tolerance levels and physiological dependency

Several days

Morphine is detected in the urine for _____ after an opiate use

Methamphetamine

Active reagent of shabu

1. Mental alertness

2. Physical capacity

3. Anorectic property

Methamphetamine increases (3):

1. Auditory hallucinations

2. Visual hallucinations

3. Paranoia

Acute psychotic syndromes of Methamphetamiine (3):

Tetrahydrocannabinol

Most potent component or psychoactive substance of marijuan

Tetrahydrocannabinol

Induces a sense of well-being and euphoria

3-5 days up to 4 weeks for chronic users

Tetrahydrocannabinol is detected in urine for chronic users for _______

Delta-9-tetrahydrocannabinol

Principal psychoactive agent of Tetrahydrocannabinol:

11-nor-deltatetrahydrocannabinol (THC-COOH)

Urinary metabolite of Tetrahydrocannabinol:

Competitive binding

Principle of immunoassay:

Gold colloid reaction pad

Ag-Ab reaction present in the _______

Positive

Absence of the changes in color for the test area:

Capillary action

Urine specimen will migrate upwards through _____

Calibrator

Specific analyte being tested

Cut-off

Expressed in ng/ml

Cut-off

Actual amount that must be reached before it can give significant effect to the competitive binding

1. Drug Ab-coupled particles

2. Drug conjugate

Test line contains (2):

Goat Ab

Control line contains

Room Temp (15-30C)

Allow the test panel to equilibrate to _____ prior to testing

Vertically

Immerse the test panel _______

10-15 seconds

Immerse the test panel vertically in the urine specimen for at least _______

100 ul (3 drops/3 ggts)

Volume of specimen:

5 minutes

The results should be read at ____

10 minutes

Do not interpret results after _____

Negative

Two lines appear

Positiev

One colored line on the control reegion

Invalid

Control line fails to appear

1. Insufficient specimen volume

2. Incorrect procedural techniques

Invalid results is due to (2):

1. Review the procedure

2. Repeat the test

3. Use a new test panel

Invalid result solution (3):

1. Ref temp

2. Room temp

2-30C

Storage of Test kits (2):

300 ng/ml

1. Barbiturates

2. Benzodiazepines

3. Cocaine

4. Methadone

5. Morphine

What is the cut-off value?

500 ng/ml

Cut-off value of methylenedioxymethamphetamine (ecstasy)

1000 ng/ml

Cut-off value of methamphetamine (shabu)

50 ng/ml

Cut-off value of tetrahydrocannabinol

1. Barbiturates

2. Benzodiazepines

3. Cocaine

4. Methadone

5. Morphine

Has a cut-off value of 300ng/ml (5):

1. Specific drug antibody-coupled particles

2. Drug protein conjugates

Reagent test contains (2):

Control line

Goat antibody is employed in the. _____

1. Centrifuged

2. Filtered

3. Allowed to settle to obtain a clear supernatant

Urine specimens exhibiting visible particles should be (3):

1. 2-8C, up to 48 hrs

2. Prolonged storage -20C

Urine specimen storage (2):