Introduction to Forensic Toxicology

PowerPoint Presentation by: Krystal Ronquillo

Timeline of Forensic Toxicology

1775_Karl Wilhelm SCHEELE (1742-1786) discovers that he could

change arsenious oxide to arsenious acid, which

in contact with zinc produces arsine. This

discovery later plays a great part in the forensic

detection of arsenic.

1813_Mathieu Bonaventure ORFILA (1787-1853), professor of medicinal

and forensic chemistry at Univ. of Paris, publishes Traite des Poisons.

Considered the father of modern toxicology. Significant contributions to development of tests for the presence

of blood in a forensic context. Credited as the first to attempt the use of a

microscope in the assessment of blood and semen stains.

*From crimezzz.net

1829_Sir Robert CHRISTISON (1797-1882), Professor of Forensic Medicine at Edinburgh publishes "Treatise on Poisons", which for many years is regarded as the standard work on toxicology in the English language.

1836_English chemist James MARSH (1794-1846) developes a test for the presence of arsenic in tissues. The "Marsh Test" is very sensitive, detecting as little as 0.02 mg As.

Continued. . .

1840_Charles LAFARGE, on a business trip in Paris, falls ill after eating a cake sent to him by his wife. Back home, he dies on January 13, 1840. His young wife Marie, widow a second time already is arrested after arsenic is found in her husband´s stomach. Yet the Marsh Test proved negative. An exhumation was ordered Mathieu Bonaventure ORFILA (1787-1853), meantime Dean of the Paris medical faculty is summoned by the court in the LAFARGE case. He applies the Marsh Test correctly and finds arsenic in the corpse. Marie LAFARGE is sentenced to life imprisonment. After serving 10 years she is released by Napoleon III in 1850 and dies 1851, still declaring her innocence.

1852_Jean Servais STAS, a chemistry professor from Brussels, and student of

ORFILA, is the first successfully to identify vegetable poisons in body tissue. In a murder trial in

1850, the male victim shows clear chemical burns in his mouth, tongue, and throat.

STAS searched for three months for the agent, and

eventually manages to isolate nicotine from the body tissues.

Using ether as a solvent, which he then evaporates to isolate the drug, he finds the potent drug. It was, in fact, the murder weapon. The man's killer had extracted it from tobacco and force-fed it to

the victim. With STAS's testimony, the killer is convicted.

1956_GOLAY first shows WCOT (wall-coated open tubular) columns for Gas Chromatography to be theoretically ideal.

1988_LEWELLEN, McCURDY, HORTON, and ASSELIN, LESLIE, McKINLEY publish milestone papers introducing a novel procedure for the analysis of drugs in whole blood by homogeneous enzyme immunoassay (EMIT).

The Black Widow of London:

Toxicology Case File-Between 1929 and 1949, Marie Besnard was said to have killed 12 people, including her husband and mother, with arsenic.

-All said victims’ bodies were exhumed and tested. Every body tested positive for high levels of arsenic.

-Since this test was the only evidence prosecutors had against Besnard, a number of problems in the case arose.

-Toxicologist Dr. Georges Beroud was deemed careless in his lab techniques, which prompted a second investigation. This one was also insubstantial>>theory suggests that through anaerobic bacteria arsenic could enter the hair of a corpse from the ground. Investigators now had to prove arsenic was not introduced to the bodies after initial burial.

-Although the second investigation also revealed signs of arsenic in the bodies, a mistake on the part of Dr. Griffon. The hair samples required exposure to radioactivity for more than 20 hours, but investigators only exposed it for 15. The results came under question again and a third time. Unable to completely prove anything, Besnard was acquitted on December 12, 1961

Acids (nitric, hydrochloric, sulphuric)

Burns around mouth, lips, nose

Aniline (hypnotics, nitrobenzene)

Skin of face and neck quite dark

Arsenic (metals, mercury, copper, etc.)

Severe, unexplained diarrhea

Atropine (Belladonna), Scopolamine

Pupil of eye dilated

Bases (lye, potash, hydroxides)Burns around mouth, lips,

nose

Carbolic acid (or other phenol) Odor of disinfectant

Carbon monoxide Skin is bright cherry red

CyanideQuick death, red skin, odor of

peach

Food poisoning Vomiting, abdominal pain

Metallic compoundsDiarrhea, vomiting, abdominal

pain

Nicotine Convulsion

Opiates Pupil of eye contracted

Oxalic acid (phosphorous) Odor of garlic

Sodium fluoride Convulsion

Strychnine Convulsion, dark face and neck

Toxin Symptom

-Toxicology is defined as the study of the adverse effects of toxins on any and all living organisms. When dealing in forensic toxicology the study delves into the application of this into solving a crime.

-Toxins are any substances that can be harmful to living organisms. They come in a number of forms (gaseous, liquid, solid, animal, mineral, and vegetable)

-Poisons are a subgroup of these toxins.

Urine Testing: The Most Commonly Used Drug Test

Urine Testing-Are the least expensive of the test methods ($7-$50 for home version). Are considered an intrusive method of testing. -Can be done at home (for example by parents) though require lab verification for accurate results. -Detect use primarily within the past week (longer with regular use). -Can be affected by abstaining from use for a period of time before the test. -Are often temperature tested to insure sample integrity.

Saliva TestingSaliva Multi Drug Test 5 tests for opiates/morphine (such as heroin), marijuana, cocaine/crack, methamphetamine & benzo. Detection in saliva tests begins immediately upon use: Marijuana and Hashish (THC): 1 hour after ingestion, for up to 14 hoursCocaine (including crack): From time of ingestion for 48 to 72 hoursOpiates: From time of ingestion for 48 to 72 hoursMethamphetamine and Ecstasy (MDMA, "Crank," "Ice"): From time of ingestion for 48 to 72 hoursBZD: From time of ingestion for 48 to 72 hoursThe test is based upon the new immunoassay technology for assuring the best accuracy. This means that you get the results in 10-15 minutes in the privacy of your home. Each test kit is packaged individually in a sterile bag.

An actual product on the market—at home saliva drug test. Although scientifically sound, the test is second rate to other more used drug tests.

Hair Analysis A new wave in drug testing.

A breakthrough in the forensic science field.

With so many loopholes in urine testing and other drug tests, hair analysis has become the most efficient and widely used form of drug testing in the work place and at home.

Steelcase administered both urine and Psychemedics' hair analysis tests to 774 applicants in order to provide an effective side-by-side comparison.

The graph to the right indicates the percentage rate of positive drug tests as compared between urine tests and drug analysis. The significant differences in percentages indicatesa greater success rate in hair testing

According to Psychmedics1. For all drugs tested, Psychemedics hair test was substantially more effective in identifying drug use, yielding an 18.0% positive rate in comparison to a rate of 2.7% for urine. 2. Because of the rapid excretion of cocaine in urine, Psychemedics was overwhelmingly more effective than urinalysis in identifying cocaine users. Sixteen times as many cocaine users were accurately identified using Psychemedics' patented technology

  STEELCASE CORPORATION COST PER POSITIVE TEST RESULT URINE VS. HAIR A further example of Psychemedics' cost effectiveness: If Steelcase had paid $50 per test using Psychemedics' hair testing and $30 per test for urine testing, the cost per positive for Steelcase would still be over $800 LESS using Psychemedics' technology.

If Steelcase were to rely on urinalysis to screen applicants for every 100 employees hired, approximately 15 would be drug users. The National Institute of Health estimates that each drug abuser costs an employer approximately $7,000 annually. Therefore, a company like Steelcase would save $105,000 per year for every 100 employees hired by pre-screening applicants with Psychemedics' hair analysis test.

To the left is a Cocaine (COC) Drug Test—a simple one step rapid immunochromatographic assay for a quick, qualitative detection of cocaine and benzoylecgonine (which has a longer biological half-life (5-8 hours) than cocaine (0.5-1.5 hours) and can usually be detected for 24-80 hours after cocaine use or exposure) as metabolites in urine.

Similarly to the right is the THC orMarijuana drug test. This test is based on the principle of the highly specific immunochemical reactions between antigens and antibodies, which are used for the analysis of specific substances in biological fluids, in this case urine.

*Most interesting about both tests is that although they primarily are simple and inexpensive, they require hair chormatography or mass spectometry testing before a positive determination is made*

Hair Chromatography testing as used in the work place will commonly find the following drugs. The molecules of some are shown below.

* Amphetamines (methamphetamines and amphetamines)* Cocaine* Opiates (heroin, morphine, and codeine)* Phencyclidine (PCP)* Marijuana (THC)

Hair analysis at APL Laboratories is performed by mirroring the federal forensic Substance Abuse and Mental Health Services Administration (SAMHSA) employment drug testing guidelines as closely as possible for a solid sample. Urine, of course, comes to us in liquid form. Hair must go through additional extraction procedures in order to release the drugs from the hair before testing. From that point on, the tests are performed almost identically to urine drug testing.

1) The sample is received in the secured Specimen Processing department to be verified for complete chain of custody, adequate sample volume and computer data entry. The sample is cut and weighed. Once cut into very small pieces, the sample is mixed to create as homogeneous a sample as possible. An internal chain of custody is created and a portion of the cutting of each sample is sent to the laboratory for screening.

*Procedure taken from APL Laboratories Continued. . .

2) The samples are washed, prepared and the drugs are extracted into a liquid form. As with urine, the screening process is performed by immunoassay techniques. Those samples which test negative are then reported as negative. If the screening process produces a suspected positive, a request is made from the laboratory to the specimen processing department to provide a new sample from the remainder of the original cuttings.

3) The second portion of the original cuttings is then washed, prepared, extracted and subjected to confirmation testing by either gas chromatography mass spectrometry (GC/MS) or gas chromatography mass spectrometry/ mass spectrometry (GC/MS/MS). If the confirmation test is negative or less than the cutoff, the sample is reported as negative. If the sample is positive the laboratory reports the sample as positive.

4) The federal guidelines require a final stage for urine -- certification review by qualified certifying scientists. At APL Laboratories, the certifiers also manually review all hair analysis tests before results are issued.

*Procedure taken from APL Laboratories

“The use of hair as a specimen to detect cocaine use was first reported in 1981 (Arnold and Püschel 1981; Valente et al. 1981). In those studies, hair samples from suspected drug abusers were analyzed by radioimmunoassay (RIA) for the cocaine metabolite benzoylecgonine (BZE) in an attempt to verify a history of cocaine use. The first gas chromatography/mass spectrometry (GC/MS) procedures for detecting cocaine in hair were not reported until 1987 (Balabanova and Homoki 1987). When this more specific technique was used, it was found that cocaine, not BZE, is the primary analyte in hair. The metabolites BZE and ecgonine methyl ester (EME), shown in figure 1, are present in such low and variable concentrations that they may result from environmental degradation of cocaine already present in hair.”

ANALYSIS OF OPIATES IN HUMAN HAIR BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

“An high-performance liquid chromatographic method with ultraviolet detection has been developed for the determination of the principal opiates (heroin, monoacetylmorphine, morphine, codeine) in human hair. An aliquot of 100 mg of hair was treated with 2 ml HCL 0.1N at 56° C overnight; it was then extracted by solid-phase extraction using Bond Elut Certify columns with the procedure provided for the opiates. Chromatography has been performed using a C18 reversed-phase column with a mobile phase consisting of water-acetonitrile (70:30 v/v) containing 0.001 M NaH2PO4 and 0.002 M sodium laurylsulphate at pH 2.1. “

ReferencesSlide (1)

Graphics:http://images.google.com/imgres?imgurl=http://www.all-about-forensic-science.com/images/forensic_microscope.jpg&imgrefurl=http://www.all-about-forensic-science.com/&h=224&w=300&sz=11&hl=en&start=5&um=1&tbnid=MZtO0FICJlAIGM:&tbnh=87&tbnw=116&prev=/images%3Fq%3Dforensic%2Btoxicology%2Bhistory%26svnum%3D10%26um%3D1%26hl%3Den

Slide (2) & (3)

Info & Graphics:http://www.crimezzz.net/forensic_history/index.htm Slide (4)

Info:http://www.crimelibrary.com/criminal_mind/forensics/toxicology/4.html

Graphics:http://news.bbc.co.uk/olmedia/1525000/images/_1525578_blackwidowspider150.jpg

Slide (6)

Graphics:http://www.dilapan.com/images/sitecheck5panel.jpg

http://www.detoxrightnow.com/product_13586.html

http://www.test-mall.com/uploads1/products/img3/5668_1117635898.jpg

http://www.amcareinc.com/mgmt/images/urine.jpg

Slide (5)

Info:http://faculty.ncwc.edu/toconnor/425/425lect14.htm

Graphics:http://www.slh.wisc.edu/news/images/red_needles.gif

http://www.forensicscience.ufl.edu/images/tubes.jpg

References

Slide (9)

Graphics:http://www.testcountry.com/images/hair.gif

http://www.columbialab.com/hairtesting/images/home.jpg

Slide (10) & (11)

Info: http://images.google.com/imgres?imgurl=http://www.workplacedrugtesting.com/images/steel1d.gif&imgrefurl=http://www.workplacedrugtesting.com/steelcas.htm&h=335&w=400&sz=33&hl=en&start=9&um=1&tbnid=EGKl409SGGPhsM:&tbnh=104&tbnw=124&prev=/images%3Fq%3Ddrug%2Btesting%26svnum%3D10%26um%3D1%26hl%3Den

Graphics:http://www.workplacedrugtesting.com/images/steel1d.gif

Slide (8)

Info:http://www.detoxrightnow.com/product_2741.html

Graphics:http://www.workplacedrugtesting.com/images/steel1d.gif

Slide (7)

Info:http://www.erowid.org/psychoactives/testing/testing_info1.shtml

Graphics:http://www.vrp.com/prodimgs/w200/9137w200.jpg

http://www.ebiinc.com/subheads/oral_chart.jpg

ReferencesSlide (14) & (15)

Info:http://www.columbialab.com/cgi-bin/web_store/web_store.cgi?cart_id=&page=/hairtesting/index.shtml

Graphics:http://images.google.com/imgres?imgurl=http://library.thinkquest.org/04oct/00206/images/marijuana.JPG&imgrefurl=http://library.thinkquest.org/04oct/00206/lo_mt_toxicology.htm&h=180&w=240&sz=46&hl=en&start=29&um=1&tbnid=BXVmjse92_N8tM:&tbnh=83&tbnw=110&prev=/images%3Fq%3Dhair%2Bgas%2Bchromatography%26start%3D20%26ndsp%3D20%26svnum%3D10%26um%3D1%26hl%3Den%26sa%3DN

http://www.hctx.net/cmp/ImageViewer.aspx?ImageID=1776

http://www.omegalabs.net/media/images/illustrations/Hair-folliculeb.jpg

Slide (13)

Info:http://www.columbialab.com/cgi-bin/web_store/web_store.cgi?cart_id=&page=/hairtesting/index.shtml

Graphics:http://www.druglibrary.org/schaffer/cocaine/CocHairFig1.gif

Slide (12)

Info:http://www.detoxrightnow.com/product_13586.html

Graphics:http://www.detoxrightnow.com/product_13586.html

Slide (17)

Info:http://www.tiaft.org/tiaft97/timetable/wednesday/034.html

Graphics:http://www.rsc.org/ej/AN/2001/b103590h/b103590h-f3.gif

ReferencesSlide (16)

Info:http://images.google.com/imgres?imgurl=http://www.druglibrary.org/schaffer/cocaine/CocHairFig1.gif&imgrefurl=http://www.druglibrary.org/schaffer/cocaine/cokehair.htm&h=252&w=450&sz=8&hl=en&start=10&um=1&tbnid=90WBkzfodWM3XM:&tbnh=71&tbnw=127&prev=/images%3Fq%3Dhair%2Bchromatography%26svnum%3D10%26um%3D1%26hl%3Den%26sa%3DG

Graphics:http://www.tau.ac.il/chemistry/amirav/cocaine.gif