society of forensic toxicologists, inc. june 2008 oxtalk

ToxTalk EditorsYale Caplan, Ph.D., DABFTVickie Watts, M.S.

Section EditorsDaniel Anderson, M.S., FTS-ABFTMatthew Barnhill, Ph.D., DABFTDwain Fuller, B.S., D-FTCBJ. Robert Zettl, MPA

SOFT 2008 Board of DirectorsPRESIDENT

Christine Moore, Ph.D., DABCCVICE PRESIDENT

Anthony Costantino, Ph.D., DABFTSECRETARY

Sarah Kerrigan, Ph.D.TREASURER

Bradford Hepler, Ph.D., DABFTDIRECTORS

Ashraf Mozayani, Ph.D., DABFTMarc LeBeau, Ph.D.Peter Stout, Ph.D., DABFTDan Anderson, M.S., FTS-ABFTDwain Fuller, B.S., DFTCB

ex officioPast President:

Diana Wilkins, Ph.D.Webmaster:

Bruce Goldberger, Ph.D., DABFT

I N S I D E T H I S I S S U E :President’s Message 2Drugs In The News 3-5Case Notes 6-10Bits & Pieces 11

Inserts:2008 Meeting Registration Worksheet2008 Preliminary Program2008 Fun Run Sign Up Form

The user friendly SOFT 2008Meeting Website can be found atwww.soft2008.org and contains the mostup to date meeting information such asthe current Preliminary Program and thedetailed information on the twelve focusoriented workshops being offered at the2008 meeting. In addition, the meetingwebsite contains local “Arizona Activi-ties” and information on the “all suites”resort, the Arizona Grand, home to theSOFT 2008 meeting. The website is up-dated routinely by meeting website de-

R E G I S T E R N O W F O R T H E

2 0 0 8 A N N U A L M E E T I N G

Amanda Gallegos of the PhoenixPolice Department Laboratory Services Bu-reau has joined Jeri Ropero-Miller of RTIInternational to coordinate the activities ofthe SSEP in Phoenix. While the SSEP targetscollege students in the regional area of theevent, all interested college students can ap-ply with equal consideration. Up to 100 stu-dents can attend SSEP. Students must pro-vide their own travel to the Arizona GrandResort and private sponsors and proceedsfrom prior SOFT Silent Auctions fund theentire day-long SSEP program. We are invit-ing these students to “Come Experience aDay-in-the-Life of a Forensic Toxicologist”.

DETAILS AT-A-GLANCE:When: Monday, October 27, 2008

Who can Apply: College Students(undergraduate and graduate)

Application Period: May 1, 2008through September 30, 2008

Applications Available from:http://www.soft2008.org/SSEP.htmlor [email protected]

Acceptance Notification:October 10, 2008

SSEP Coordinators:Jeri Ropero-Miller, Ph.D.RTI International919-485-5685; [email protected]

Amanda GallegosPhx. Police Dept., Lab. Services602-262-6197;[email protected] Gallegos

veloper, Douglas Kramer (a newSOFT member), and contains manylinks to related sites for this meeting.We highly recommend that you log infrequently and see “What’s New”.

Use this site to register andselect desired workshops before theyreach maximum capacity. AttendingSOFT annual meetings is the best wayto learn the latest scientific develop-ments in forensic toxicology, plus en-joy the hospitality and culture of eachdistinctive host city.

Jeri Ropero-Miller

Society of Forensic Toxico logists , Inc .Volume 32, Issue 2

June 2008

TOXTALK®

S . O . F. T. S T U D E N T E N R I C H M E N T P R O G R A M

Sunday, October 26, 2008

Registration Opens (9:00am-6:00pm)

NLCP Training (2:00pm-6:00 pm)

Dinner on your own

Monday, October 27, 2008

Continental Breakfast (7:00am-8:30am)

Registration (7:00am-6:00pm)

SOFT Workshops (8:00am-5:00pm)

SOFT Student Enrichment Program(8:00am-5:00pm)

ABFT Exam Committee

SOFT-AAFS Drugs and DrivingCommittee

Tier-One Exhibitors Hospitality(6:30pm-8:30pm)

Tuesday, October 28, 2008



SOFT Workshops (8:00am-5:00pm)

SOFT Board Meeting (7:00am-noon)

ABFT Exam

Thursday, October 30, 2008

SOFT Fun Run/Walk (6:30am-8:00am)



Exhibits open (7:30am-1:30pm)

Exhibitor Feedback Mtg (8:00am-9:30pm)

Scientific Session (8:30am-noon)

Lunch with Exhibitors (noon-1:15pm)

Exhibits Breakdown (1:30pm-3:30pm)

Scientific Session (1:15pm-2:30pm)

SOFT Business Meeting (3:00pm-5:00pm)

ABFT Certificate ReceptionWine & Cheese (5:00pm-6:00pm)

Presidents Banquet and Masquerade Ball(6:30pm-11:30pm)

Friday, October 31, 2008


Scientific Session (9:00am-noon)

NSC Executive Board (1:00pm-3:30pm)

Greetingsfrom Southern Cali-fornia….It’s over90oF today – life isgood!

As timeseems to go more

quickly, the meeting planning for SOFT2008 is well underway and as I write, theregistration page has just “gone live”.Please make sure you register early sincethe workshops being offered are of a veryhigh quality and space will be limited –don’t miss out.

The Plenary Session focuses onphenethylamines and the speaker is theworld-famous Dr. Alexander Shulgin,who himself discovered many of thesecompounds. Dr. Shulgin has been active inthe field of drug synthesis, co-authoringthe books PiHKAL (Phenethylamines IHave Known and Loved: A ChemicalLove Story) and TiHKAL (Tryptamines IHave Known and Loved: The Continua-

ABFT Accreditation Committee

ABFT Board Meeting

Exhibits Setup (noon-5:00pm)

Exhibits Open (6:30pm-8:00pm)

Welcoming Reception (6:30pm-8:00pm)

Elmer Gordon Forum (8:00pm-10:00pm)

Nite Owl Reception (10:30pm-12:30am)

Wednesday, October 29, 2008



AAFS Steering Committee (9:00am-10:00am)

Exhibits open (7:30am-3:30pm)

Opening Ceremonies Plenary Session

Scientific Sessions (8:30am-noon)

Lunch with Exhibitors (noon-1:15pm)

DFSA Committee Meeting (noon-1:15pm)

Scientific Sessions (1:15pm-5:00pm)

Exhibitor’s Happy Hour (5:00 pm-6:30pm)

“Sunset at the Oasis” Poolside Reception(7:00 pm-10:00 pm)

S . O . F.T. 20 08 A N N U A L M E E T I N GPhoenix, Arizona, October 27-31, 2008

Hosts: Vickie Watts / Norman WadeSite: Arizona Grand Hotel (formerly the Pointe South Mountain)

P R E L I M I N A R Y P R O G R A M

tion), which discusses various aspects ofpsychoactive drugs….should be interest-ing!

The Editor for the Special Issueof the Journal of Analytical Toxicology2008, Dan Anderson, reports that he re-ceived 33 submissions, all of which arecurrently under review/revision. Thatstatistic alone shows the importance ofour annual meeting, and leads into an-other SOFT Board initiative.

Each year the Special Issue isfull of excellent research and the SOFTBoard wanted to create an Award to re-flect the best published article. TheAward, to be entitled “Experimental De-sign and Impact on Toxicology” (EDIT),will recognize the first author of the paperthat year, which, in the opinion of thejudges, shows excellent scientific experi-mental design and has a wide impact onour field. The award will begin in 2009,and to ensure the Special Editor is notoverwhelmed, three external judges will

be asked to recommend the winner fromaccepted manuscripts prior to publicationin the Special Issue. We are working withTinsley Preston at JAT and Bruce Gold-berger, Editor JAT to determine the pre-cise logistics for 2009.

At the moment though, I amdelighted to announce that the judges forthe premier EDIT Award will be Drs.Edward Cone, Amanda Jenkins and HansMaurer. I am sure you will agree thatbetween them, these toxicologists have avast knowledge of experimental designand extensive publication history. TheSOFT Board is excited about the creationof this new initiative to recognize ourscientists, and we encourage you to sub-mit your research to our Special Issue.

Finally, just a reminder that wewill be voting on increasing dues by $10per year at the 2008 Business Meeting.

I thank you for your loyal mem-bership and wish you all a very relaxingsummer. Hope to see you in October.

P R E S I D E N T ’ S M E S S A G E

B Y C H R I S T I N E M O O R E , P H . D . , DAB C C

Volume 32, Issue 2

Submitted by Section Editor, Dwain Fuller, D-FTCB, TC-NRCC

D R U G S I N T H E N E W S

R I C I N

Castor Beans

Castor beans come from thecastor bean plant (Ricinus communis)which has been cultivated for centu-ries, primarily for the oil produced byits seeds, or beans. The Egyptiansburned castor oil in their lamps morethan 4000 years ago. Castor oil, is per-haps best known to the lay person as apopular stimulant laxative of the early20th century. As a child of the 60’s, Ihave never experienced the joys of cas-tor oil, but merely broaching the sub-ject in my workplace initiated a torrentof stories of the horrors of taking cas-tor oil. The aversion to the medicinaluse of castor oil seems to lie in its tasterather than its effects.

There are many non-medicaluses of castor oil as well. In the UnitedStates, castor oil has been used in air-craft lubricants, hydraulic fluids, syn-thesis of soaps, linoleum, printer’s ink,nylon, varnishes, and in the manufac-ture of explosives. The seeds, stemsfrom the pressing of castor oil containsabout 5% ricin by weight. While it hasbeen estimated that as few as four toeight castor beans would be toxic oreven fatal to an adult human, the fact isthat unless the seed coat is broken,such as by chewing, castor beans willlikely pass through the body with no illeffects. Perhaps in testimony of itsrelatively benign nature, when not pur-posely abused some varieties of thecastor bean plant are often grown asornamentals. They are best adapted tothe soils and climate of southeasternKansas and Missouri, southern Illinoisand Indiana, as well as Tennessee,Kentucky, and parts of Oklahoma andTexas.

While castor beans do not ap-pear to pose too much of a threat, ricin

Castor Bean Plant

Please send interesting “Drugs In The News” to Section Editor, Dwain Fuller [email protected]

itself, however, is a potent toxin. Theestimated toxic dose for an adult isless than 1 milligram if inhaled orinjected. There is no known antidotefor ricin poisoning. Instead onlysymptomatic and supportive treatmentis available and long-term organ dam-age is likely in survivors. The symp-toms of ricin poisoning are dependenton the route of administration. If in-haled, the symptoms are: respiratorydistress, fever, cough, nausea, andtightness in the chest. These are fol-lowed by pulmonary edema, hypoxia,cardiac arrhythmia, and death. If in-gested, the symptoms are: vomiting,diarrhea, perhaps becoming bloody,followed by severe dehydration.Within several days the person’s liver,spleen and kidneys may fail, leadingto death.

Ricin was evaluated by theUnited States for its potential as amilitary weapon during World War I.At that time it was being consideredfor use either as a toxic dust or

On February 14th, 2008, whatseemed like a routine call to a LasVegas 911 operator from a man suf-fering from respiratory problems be-came an incident of far-ranging sig-nificance. At last report, the man,now identified as Roger Von Bergen-dorff, a down-and-out graphic artist,was in critical condition in SpringValley Hospital, and by at least onereport, comatose.

The cause of his conditionwas not discovered until two weekslater, however. At around 2:30 p.m.on February 28th, Thomas Tholen, acousin of Von Bergendorff, wascleaning out the Extended Stay Amer-ica room that Von Bergendorff hadrented, and from which he was nowbeing evicted, when he came across asmall vial. Mr. Tholen took the vialdown to the motel’s front desk whichset in motion a massive police,hazmat and Homeland Security re-sponse. The content of the vial wasdetermined to be ricin.

Ricin, as you may be familiar,is a toxin derived from castor beans.Its mechanism in the body is to inhibitprotein synthesis. As a result of thisinhibition, the cells are deprived ofessential proteins and die. Thus ricinis a systemic poison affecting multiplesystems of the entire body.

Volume 32, Issue 2

Ricin A - blue, Ricin B - redPellet recovered from Georgi Markov

ToxTalk

as a coating for bullets or shrapnel.Due to the technological limitations ofthe time the dust-cloud concept couldnot be adequately developed and thebullet/shrapnel coating concept wouldbe a violation of the Hague Conventionof 1899. During World War II theUnited States and Canada studied thepotential for the use of ricin in clusterbombs, but concluded that due to thenecessity to aerosolize it as a dust, itwas no more economical than phos-gene.

Perhaps the most interestingchapter in the history of ricin is theincident of Georgi Ivan Markov.Georgi Markov was a Bulgarian-bornnovelist and playwright. Markov en-joyed a privileged existence in Bulgar-ian society even though his father wasconsidered a “class enemy” of thecommunist party. This all ended, how-ever, in 1969 when he defected to Italyafter learning that his latest play hadangered the government and put him atrisk. After his defection he was ac-cused and convicted by Bulgarian au-thorities, in absentia, of being a traitor.By 1971 Markov had immigrated toBritain where he became a broadcastjournalist and commentator for theBBC. In, June of 1975, he began con-tributing programs to the CIA-funded,Radio Free Europe, where his weeklyshows were sharply critical of Bulgar-ian bureaucrats and communist partyofficials, especially, party leader, To-dor Zhivkov.

In early 1978, Markov beganreceiving death threats. In the last callin August of 1978, Markov was toldthat he would die of natural causes,killed by a poison the West could notdetect nor treat. Two weeks later onSeptember 7th, Markov parked his carin a parking lot on the south side ofWaterloo Bridge in London. This washis usual parking spot where he wouldcatch the bus to BBC headquarters.While waiting at the bus stop, Markovfelt a sharp prick in the back of hisright thigh. When he turned around he

saw a gentleman bending over to pickup a dropped umbrella. The man said“I’m sorry” in a foreign accent,promptly hailed a cab, and left.

Markov, although in pain, con-tinued on to work where he told hiscolleagues what had happened. On theback of his right thigh was a swollenpimple-like wound. That evening hedeveloped a high fever and by the nextday was having trouble talking. Hewas admitted to the hospital where hewas initially treated for septicemia, butover the next few days Markov beganto have bloody vomit and kidney fail-ure. On September 11, 1978, his heartfailed. Georgi Markov was dead.

At autopsy it was determinedthat Markov’s lungs were full of fluid,his white blood cell count was ex-tremely high, his liver was damaged,and his lymph nodes, intestines andheart were riddled with small hemor-rhages. A large portion of tissue wasremoved from around the wound areaon the back of Markov’s right thigh.In this tissue examiners discovered asmall pellet 1.52 mm in diameter com-posed of platinum and iridium. Thepellet was eventually determined to bea watch bearing, but in this pellet weretwo 0.34 mm holes bored at right an-gles to each other, forming an Xshaped well inside. Due to the extremehardness of this material it was sur-mised that such precision machiningcould only have been done by a sophis-ticated laser process known as “sparkerosion.”

Although no poison was de-tected in the pellet, nor in Markov’s

body, it was surmised due to strongcircumstantial evidence, based onMarkov’s symptoms, postmortem pa-thology, and intelligence informationregarding the Soviet Union’s involve-ment in research of the use of ricin as aweapon, that Markov did in fact die ofricin poisoning administered via thepellet propelled from a gas powereddevice disguised as an umbrella. Al-though the Soviet Union has deniedany connection to the incident, KGBdefectors, Oleg Kalugin and OlegGordievsky, have since confirmed theSoviet Union’s involvement.

Ricin consists of two distinctprotein chains with a molecular weightaround 30 kDa each. Ricin A is an N-glycoside hydrolase that targets anddepurinates an adenine base in the 28SrRNA molecule of the ribosome, re-sulting in an inhibition of protein syn-thesis. Ricin B is a lectin that bindsgalactosyl residues and is important inassisting ricin A’s entry into a cell bybinding the cell surface component. Itis the presence of both of these chainsthat renders ricin so toxic. Manyplants such as barley have the ricin Achain but not the ricin B chain. Sincepeople do not get sick from eating evenlarge quantities of such products, thericin A chain is relatively harmlesswithout the ricin B chain present.

D R U G S I N T H E N E W S ( C O N T I N U E D )

D R U G S I N T H E N E W S ( C O N T I N U E D )

Ricinine

SU NS H IN E / R IE DE RS S I LE NT AUC T IO NSOFT sponsored a memorial

event during the 2006 Austin meetingto honor the recent passing of twoillustrious leaders in forensic toxicol-ogy, Dr. Irving Sunshine, and Dr.Frederick Rieders. The event wasnamed the “Sunshine / Rieders SilentAuction” and has since become anannual event that meeting attendeeslook forward to. A wide array ofitems are donated by exhibitors andindividuals, then displayed with bidsheets, tracking names and bids. At a

designated time, bidding closes andwinners can pay and pick up their treas-ures. Not only is the auction a fun tradi-tion, but complete proceeds benefit stu-dents interested in forensic toxicologythrough the SOFT Student EnrichmentProgram. Since Dr. Sunshine and Dr.Rieders focused their energy on aca-demic encouragement in this field, it isthought to be an appropriate way to ac-knowledge their lifetime contributionsand continue their legacy of promotingeducation in forensic toxicology.

SOFTMember, LaurieTobler has gener-ously volunteeredto coordinate the2008 Sunshine /Reiders SilentAuction. Anyonewishing to donateitems, big or small, should contact:

Laurie Tobler at [email protected]

Ricins may have therapeuticuse in the battle against cancer. It istheorized that Ricin could be linked toa monoclonal antibody to target ma-lignant cells recognized by the anti-body. It has also been postulated thatone may be able to use the ricin Bchain as a vehicle to effectively de-liver antigens into cells thus greatlyincreasing their immunogenicity.

The fact that ricin is a com-plex protein of extremely high mo-lecular weight presents an unusualproblem for the forensic toxicologist.On one hand, these characteristics ofricin make it relatively well-suited fordetection by a properly designed im-munoassay, and many of these havebeen described in the literature. How-ever due to its high molecular weightand complex structure, confirmationby electron-impact GC/MS or LC/MS, the standard tools of the forensictoxicologist, is not possible.

Although not fully exploredin human subjects, the literature sug-gests that a biomarker of ricin expo-sure, ricinine, may be easily detectedby standard solvent extraction andGC/MS or LC/MS technology.Ricinine is not a metabolite of ricin,but rather a biomarker that derivesfrom the same source as ricin, the cas-tor bean plant. Thus confirming thepresence of ricinine in biological flu-

ids that have screened positive for ricinby immunoassay greatly enhances evi-dence of ricin exposure or poisoning.Ricinine (C8H8N2O2) has a molecularweight of 164 daltons and thus is quiteamenable to analysis by routine GC/MS or LC/MS technology.

Update:Near the completion of this

article it was reported that Roger VonBergendorff had recovered and uponhis release from the hospital on April16th, 2008 was arrested and chargedwith possession of a deadly toxin. Mr.Von Bergendorff allegedly admittedmaking ricin in what he described as an“exotic idea” to harm his enemies.Found, among other clandestine items,at a self-storage rented by Von Bergen-dorff, was a safe containing a ricincontaminated mortar and a drawingmade by Von Bergendorff of an injec-tion device disguised as a pen.

References:

1. Abigail Goldman, Meet the myste-rious Roger Von Bergendorff, LasVegas Sun, March 5, 2008

2. E.S. Oplinger, E.A. Oelke, A.R.Kaminski, S.M. Combs, J.D. Doll,R.T Schuler, Castorbeans, Alterna-tive Field Crops Manual, Univer-sity of Wisconsin-Extension, Coop-erative Extension, University ofMinnesota: Center for Alternative

Plant & Animal Products and theMinnesota Extension Service.

3. Ricin, Wikipedia, http://en.wikipedia.org.wiki.Ricin, ac-cessed 3/17/08

4. Case File: Umbrella Assassin,Secrets of the Dead, http://www.pbs.org/wnet/secrets/case_umbrella/p_index.html,accessed 4/14/08

5. Facts About Ricin, CDC, March5, 2008, http://www.bt.cdc.gov/agent/ricin/facts.asp, accessed3/17/08

6. Ricinus communis (Castor bean),http://www.ansci.cornell.edu/plants/castorbean.html, accessed4/15/08

7. Darby SM, Miller ML, AllenRO. Forensic determination ofricin and the alkaloid markerricinine from castor bean ex-tracts. J Forensic Sci 2001;46(5):1033-1042

8. Johnson RC, Lemire SW,Woolfitt AR, Ospina M, PerryKP, Olson CT, Barr JR. Quantifi-cation of Ricinine in Rat and Hu-man Urine: A Biomarker forRicin Exposure. J Anal Toxicol2005;29:149-155

9. Erin Alberty, Nevada man ar-rested after ricin found in W.Jordan storage unit. The SaltLake Tribune, April 17, 2008

Volume 32, Issue 2

Submitted by Section Editor, Matthew Barnhill, Ph.D., DABFT

Please send interesting “Case Notes” to Section Editor, Matthew Barnhill, Ph.D.at [email protected]

Introduction:In 2006, a sample of blood

(along with several others) was submit-ted to the laboratory at Northern TierResearch for routine toxicological test-ing by a local enforcement agency.This testing included screening fordrugs that may have impaired the abil-ity of a driver to control his / her motorvehicle. The sample, at that time wastested by immunoassay after solidphase extraction using Clean Screen TM

DAU columns. The sample was alsoscreened for drugs of abuse by gaschromatography-mass spectrometry ona HP 5890/5973 MSD (opiates, am-phetamines, cocaine, cannabinoids)with negative results using the sametype of solid phase columns for extrac-tion of the sample. Analysis of theblood sample for prescription drugs bysolid phase extraction and gas chroma-tography- mass spectrometry revealedthe presence of butalbarbital, diaze-pam/nordiazepam and propoxyphene/norpropoxyphene: ( 3.1 mg/L, 0.5 mg/L, 0.7 mg/L, 0.29 mg/L, norpropoxy-phene (positive), respectively). Fol-lowing review of the analytical find-ings a report was filed.

In March 2008, as part of amethod development project evaluat-ing new solid phase/ liquid chromatog-raphy columns for the use in forensictoxicology, the blood sample was re-trieved from long term storage where ithad been kept in a refrigerated condi-tion. The sample of blood was ana-lyzed along with several others in ablind trial to see how both types of col-umns (solid phase and liquid chroma-tography) would perform using real

sample types. Prior to analysis of thesamples, no information regarding thedrugs was known. In analyzing thisparticular sample, a benzodiazepinescreen was performed in which nordi-azepam and oxazepam were con-firmed by LC-MSMS. A basic drugscreen was also performed on the LC-MSMS in which the propoxyphene/norpropoxyphene were also con-firmed. In addition to those drugs,methaqua-lone was also detected. TheMRM values for methaqualone(251.2-> 132.1, 251.2-> 91.0, respec-tively) were set up within the programprior to analysis. Confirmation wasachieved by comparison with an unex-tracted sample of methaqualone.

Experimental (LC-MSMS):For screening purposes, 1 mL

of sodium acetate buffer (1 M, pH 4.5)was spiked with propoxyphene-d11and diazepam-d5, respectively as in-ternal standards to which 1 mL of thesample blood was added. The mixturewas diluted with a further 2 mL of thebuffer, vortexed and centrifuged. Thesample was applied to a SSCCXHsolid phase column obtained from

United Chemical Technologies, Inc.The column washed with distilled wa-ter and an acetonitrile/acetate buffermix (5: 95 v/v), respectively then driedand eluted with 3 mL of a mixture ofethyl acetate/acetonitrile (78:20 ) con-taining 4% ammonium hydroxide byvolume. The eluate was evaporated todryness and the residue dissolved inmethanol (100 µL) prior to analysis byLC-MSMS with a 5 µL injection vol-ume. Separation of the analytes wasperformed using a SelectraTM Phenylcolumn ( 50 x 2.1 (3µm) using a gradi-ent mobile phase program of acetoni-trile / 0.1 % formic acid. Detection wascarried out using an API2000 massspectrometer in positive MRM mode.

For quantification of themethaqualone in the blood sample, thesame extraction procedure was per-formed except for the replacement ofdiazepam-d5 by methaqualone- d7.Negative samples were spiked withmethaqualone and methaqualone-d7.

Chromatogram of screenshowing: Methaqualone/Norpropoxyphene/

Propoxyphene (Propoxyphene-d11)(Upper trace)

Methaqualone (MRM 251.2->132.1) (Lower trace)

Results and Discussion:

From the analysis of the sam-ple, methaqualone was found to be <0.001 mg/ L (1 ng/ mL). A search/review of the stored gas chromatogra-phy-mass spectrometry computer filedid not reveal the presence of themethaqualone in the earlier analysis,

Jeffery Hackett and Michael Coyer, Northern Tier Research, Mayfield, PA

Structure of Methaqualone

C A S E N O T E S

Volume 32, Issue 2

C A S E N O T E S # 1 : M E T H A Q U A L O N E I N A S T O R E D D U I D C A S E

XIC of +MRM (87 pairs): 278.3/91.1 amu from Sample 2 (blood032008b) of march202008f.wiff (Turbo Spray) Max. 185.7 cps.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19Time, min

0.0

1.0e4

2.0e4

3.0e4

4.0e4

5.0e4

6.0e4

6.7e4

Inte

nsity,

cp

s

12.53 15.871.19 18.372.34 6.32 14.417.28 12.07 13.652.663.15 3.73 8.70 19.565.53 10.61

XIC of +MRM (87 pairs): 251.2/132.1 amu from Sample 2 (blood032008b) of march202008f.wiff (Turbo Spray) Max. 9385.7 cps.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19Time, min

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

Inte

nsity,

cp

s

10.47

pact upon the drivers ability to controlthe motor vehicle. Although somestudies have been published regardingthe stability of drugs stored in bloodsamples 5-6, it is not known by thisparticular laboratory exactly howmethaqualone behaves when storedunder refrigeration for a long periodof time. It is thought that the diaze-pam originally present may have bro-ken down to its metabolites during thetime the blood sample was stored.

This use of LC-MSMS hasshown that older samples may havecontained drugs that were not detectedpreviously.

References:1. R.C. Baselt, Disposition of Toxic

Drugs and Chemicals in Man, 5th

Ed. Biomedical Publications, Fos-ter City, CA 2000, pp.536-538

2. Clarke’s Analysis of Drugs andPoisons, Pharmaceutical Press,A.C. Moffat, M.D. Osselton, andB.Widdop , (Eds). Pharmaceutical

Press, London, UK (2004)pp1237-8.

3. H.H McCurdy, E.T. Solomons, andJ.M.Holbrook. Incidence ofmethaqualone in driving-under-theinfluence (DUI) cases in the stateof Geogia. J.Anal. Toxicol. 5: 270-4 (1981)

4. P. Krantz and Wannenberg. Occur-rence of barbiturate, benzodi-azepine, meprobamate,methaqualone and phenothiazine incar occupants killed in traffic acci-dents in the south of Sweden. Fo-rensic Sci. Int. 18: 141-7 (1981)

5. F.T. Peters. Stability of analytes inbiosamples- an important issue inclinical and forensic toxicology?Anal. Bioanal. Chem. 388: 1505-19 (2007)

6. M.D. Robertson and O.H. Drum-mer. Stability of nitrobenzodi-azepines in postmortem blood. J.Forensic Sci. 43: 5-8 (1998)

although the other reported drugs werepresent. A review of the other samplesanalyzed by solid phase extraction/LC-MSMS before and after the presentsample in the sequence list did not re-veal the presence of methaqualone.

Methaqualone is a sedative anda hypnotic, first synthesized in 1951and entered into the market place in1956, but due to its misuse the drugwas removed from use in the US in19841. Therapeutic levels ofmethaqualone are reported to be 0.4 to5 mg/ L (plasma). 2 Toxic levels ofmethaqualone in post mortem bloodhave been reported to be in the range5-42 mg/ L2.

Several reports indicating thedetection and quantification ofmethaqualone in driving cases havebeen published in the 1980’s 3-4 but thisis the first time that we have encoun-tered this particular drug. The level ofthe methaqualone detected, althoughsub therapeutic, may have in combina-tion with the other drugs had some im-

ToxTalk

C A S E N O T E S # 1 ( C O N T I N U E D )

Tim Grambow and Laurie Shacker, Toxicology Department, SC Law Enforcement Division

Figure 1: HS-GC/FID Chromatogram

Figure 2: HS-GC/MS Chromatogram

Figure 3: Mass Spectrum of 1,1-difluoroethane

Introduction:The abuse of inhalants

in order to achieve a “high” isnot a new problem. Four majorclasses of abused inhalants havebeen identified. They are anes-thetic gases (including nitrousoxide), industrial solvents(including hydrocarbons), aero-sol propellants (includingfluorocarbons) and organic ni-trites (amyl or butyl).1 Absorp-tion of inhaled chemicals intothe lungs leads to a quick distri-bution into the blood and subse-quently the brain. Variousmechanisms of action includeoxygen displacement and inter-

actions with ionchannels. Theuser experiencesintoxication andeffects that aresimilar to thoseof alcohol in-cluding slurredspeech, de-creased motorskills, euphoria,and dizziness.2

The quick onsetof action com-bined with easeof obtaining

these compounds makes inha-lants popular alternatives toother drugs. As can be ex-pected, using inhalants can im-pair one’s ability to safely oper-ate a motor vehicle.

Trends:

In 2007 the South Caro-lina Law Enforcement Division(SLED) received approximately1,750 toxicology requests forDUI cases and 530 traffic fatali-ties. A volatile analysis wasperformed on submitted sampletypes which included blood

C A S E N O T E S # 2 : I N H A L A N T S A N D D R I V I N G I N S O U T H

C A R O L I N A : A V O L AT I L E S I T U AT I O N

Volume 32, Issue 2

C A S E N O T E S # 3 : H O W M A N Y D R U G S D O E S I T T A K E T O P R O V E

I M PA I R M E N T ?

Prosecutors in our state liketheir toxicology neat. They want tohear "The driver was definitely im-paired. With that drug level, you areintoxicated. No question." They getirritated when they are told,"Maybe...it depends on the circum-stances...it depends on the subject'smedical history...I can't say that couldnever be true...." But sometimes youget a case where the toxicologicalfindings seem to be pretty solid, theconclusions pretty straightforward,and you know your prosecutor is go-ing to be happy. For instance, thisone:

Scenario: A fatal crash oc-curred at 12:45 pm in a Detroit suburb.A driver in an SUV broadsided a van atan intersection, killing the van's elderlydriver. Toxicological examination ofthe SUV driver's blood showed 566 ng/mL diazepam, 1081 ng/mL nordiaze-pam, 8 ng/mL hydrocodone, 135 ng/mL methadone, a methadone metabo-lite (not quantified) and 17 ng/mL 11-COOH-THC (no parent THC).

This driver had a lot prescrip-tion drugs on board, not to mention thecannabinoid. The prescriptions are allwithin therapeutic ranges, but thetherapeutic ranges for diazepam and

methadone are extremely broad (20 -4000 ng/mL and 75 - 1100 ng/mL, re-spectively1); and, as any good toxicolo-gist knows, "therapeutic range" doesnot equal "no effects." Even singledoses of benzodiazepines can impairvigilance2,3, and these levels of diaze-pam and nordiazepam are more reflec-tive of multiple doses or chronic use.And then there's the cannabinoid: amoderate amount, in our lab's experi-ence; and although no THC was found,THC resides in the brain after leavingthe blood4, where it presumably exertsits effects for some time further. Theeffects of THC can last up to 24 hours

Michele Glinn, Ph.D., Michigan State Police Toxicology Laboratory, Lansing, MI

ToxTalk

and/or urine dependent on the individ-ual case and South Carolina DUI law.No inhalants were reported in thesecases until July. Two cases with inha-lants were reported in July along with1 case each in August, September andDecember. In all 5 of these cases theinhalant that was reported was 1,1-difluoroethane. This compound ismost often found as an aerosol propel-lant, a component of cannedair duster and is being phased out as arefrigerant (Freon-152a).

Six more instances of inha-lants have been reported in DUI casesthrough April of 2008. In these cases1,1-difluoroethane, isoflurane andtoluene were detected. Isoflurane is avolatile anesthetic (Forane) whiletoluene is a petroleum based solvent.In 5 of the 8 cases involving 1,1-difluoroethane an indication of inhal-ing canned duster was given in thecase history (several naming the brandsuspected). In a single case, a canisterwas also submitted as evidence. Noindication of inhalants was given forthe cases involving isoflurane or tolu-ene.

Detection:

All samples were initially ana-lyzed using headspace gas chromatog-raphy with an FID detector. The HS-GC/FIDs used utilize either a RestekBAC-2 or a Carbowax-20M columndepending on the individual instru-ment. A secondary analysis was runusing HS-GC/MS with a Restek BAC-1 column. All results were qualitativeonly.

Discussion:

The effects of inhalants maketheir detection as important as otherimpairing drugs for DUI and trafficfatality cases. The detection of inha-lants in 11 cases over the last 10months compared to zero for the previ-ous 6 months shows a large increase inusage by drivers. The two largest limit-ing factors in the detection of theseinhalants is time of sample collectionand instrumentation. First, the tissuesolubility of various gases will affectthe time window for collection. Nitrousoxide has a half-life of 5 minutes ver-sus toluene which has an initial aver-age half-life of 4.5 hours.3 Very short

half-lives require the sample to be col-lected soon after the supposed time ofimpairment otherwise the gas may beeliminated. Second, the specificity ofthe detector during the analysis affectsthe ability to identify various com-pounds. Mass spectrometry allows forexplicit identification of a compoundbased on fragmentation patterns. Theability to reference a library of massspectrums instead of unknown reten-tion times allows proper standards tobe used. This is advantageous sincethere is such a wide range of inhalantsand often little or no case history issubmitted.

References:

1. Basic & Clinical Pharmacology;Katzung, B., Ed. McGraw-Hill:New York, 2004.

2. Inhalant Abuse; National Instituteon Drug Abuse Research Report(NIH Publication Number 05-3818). Revised March 2005.

3. R.C. Baselt. Disposition of ToxicDrugs and Chemicals in Man, 7th

ed. Biomedical Publications, Fos-ter City, CA 2004.

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Volume 32, Issue 2

It is possible that without thedrugs on board, the defendant wouldhave been able to react to the victimcutting him off, cell phone or no;however, this is mere speculation.The honest toxicologist cannot dis-pute the defense's likely positionsthat the driver is a tolerant user of theabove prescribed medications, thatacute effects of the cannabinoids hadlikely resolved by the time of theaccident, and that trained medicalwitnesses would be expected to no-tice or document if there was evi-dence of impairment. In summary,there is no evidence besides the labreport that the driver was impaired,and a fair amount of circumstantialevidence that he wasn't.

I could not conclude that thisdriver was impaired, and the resultwas one more dissatisfied prosecutor.Beyond that, it was a lesson in not tojump to conclusions based on the labreport, and to consider all the cir-cumstances of the case before formu-lating an opinion.

References:1. Winek C. et. al. Forensic Sci-

ence International.(2001):122(2):107-123.

2. Kozena L. et al., Psychopharma-cology (1995):119:39-46.

3. Nakazono K. et al, YakugakuZasshi (Pharmaceutical Soc. ofJapan), (2005):125(3):307-314.

4. Mura P. et al, J. Analytical Toxi-cology (2005): 29:842 (letter).

5. Huestis M.A, Forensic ScienceReviews (2002), 14(1/2):15-59.

6. Baselt R., Disposition of ToxicDrugs and Chemicals in Man,6th Ed., 2002, Biomedical Publi-cations, Foster City, CA.

7. Drummer O.H., Forensic Sci-ence Reviews (2002), 14(1/2):1-14.

8. Stout P.R., Forensic Science Re-views (2003), 15(1):29-60.

depending on the ability tested5, cer-tainly past the time where it might nolonger be detectable in blood.

The lab report raises the sus-picion that the driver was impaired,probably seriously impaired. Onemight expect the rest of the circum-stances of the case to support thisopinion. Only they don't, exactly.

First, there are the observa-tions of the witnesses. The crash oc-curred because the victim's van cut offthe defendant's SUV at the intersec-tion. The elderly driver of the vanhad a history of bad driving and hadhad his license suspended in the pastbecause he had been involved in somany accidents. The defendant wasnot speeding and hit the brakes beforethe crash, as shown by the skid markshe left behind. No bad driving on hispart was seen by anyone. Witnesseswere unanimous that the crash wasthe fault of the victim, and the policereport reflected as much.

The police and EMTs werecalled to the scene and transportedboth drivers to the hospital. The de-fendant was treated, observed andinterviewed by a variety of trainedpersonnel. None reported any appar-ent impairment. Hospital reportsshowed the defendant to be alert, con-scious, and oriented. The police re-port specifically mentioned that noimpairment was observed. No fieldsobriety tests were done, and it seemsthat none were thought necessary.

Blood was sent to the Michi-gan State Police for toxicologicalanalysis as a matter of procedure.The officers were stunned by the re-sults.

But when the defendant wasshown a copy of the lab report, headmitted to all of it. He had prescrip-tions for Dolophine, Valium and Lor-cet. He took them routinely, and hadbeen doing so for at least six months.His normal dose was 3-4 tablets ofValium per day for muscle spasms.He had taken 2 Lorcet and 1/2 Dolo-phine early the day of the accident,

C A S E N O T E S # 3 C O N T I N U E D :and admitted to smoking marijuanaabout 11 pm the night before.

There was no obvious evidenceof any impairment on the part of thedefendant. Still, one could hypothesizethat his divided attention abilities andreaction time could have been compro-mised with so many drugs in his sys-tem. He might have been able to reactmore quickly to an unexpected event ifhe hadn't been so medicated. But thedefendant also admitted to being on hiscell phone as he was driving. Foolish,maybe, but not illegal. If he was notable to respond quickly enough to anunexpected event, was it the drugs...orwas it the distraction of the cell phone?

The drug levels seen here areconsistent with the defendant's story.Steady-state blood levels of chronicdiazepam and methadone users canreach 1500 ng/mL and 1000 ng/mL re-spectively1,6. Tolerance to the sedativeeffects of both drugs develops, and be-cause of this, blood levels alone are nota good indication of impairment7,8. Thelevels of hydrocodone are also quitelow, consistent with a dose severalhours beforehand (therapeutic range, upto 250 ng/mL1); the acute effects wouldbe expected to be in decline. And if thedriver had smoked marijuana the eve-ning before, most of the acute cannabiseffects would have returned to baselineby the time of the accident5. Couldthere be residual effects? Possibly.Could there be a combination effectfrom everything? Probably. But thereis no manifestation of such the toxicolo-gist can point to and say, "That showshe was impaired." Bad driving? None.Cause of the crash? Victim's fault.Witness observations? Not impaired.Field sobriety testing? None. Reactiontime deficit? Possible distraction fromthe cell phone. The only evidence forimpairment is the lab report, and againstthat are the statements of several profes-sional and lay witnesses that nothing inthe driving or subject's behavior indi-cated impairment and that the victim'sbehavior, not the defendant's, was thecause of the accident.

A . A . F. S . / S . O . F. T.J O I N T D R U G S &

D R I V I N G

C O M M I T T E E

N AT I O N A L S A F E T Y

C O U N C I L — C O M M I T T E E

O N A L C O H O L A N D

O T H E R D R U G S

A . A . F. S .N E W S

—T O X I C O L O G Y S E C T I O N

The NSC/COAOD last met inFebruary of 2008 in San Antonio,Texas. The committee officers for thisyear remain: Jerry Landau, Chair,Mack Cowan, Vice Chair, and LauraLiddicoat, Secretary; as well as otherstanding technical subcommittee chairsand co-chairs.

The next executive committeemeeting of the NSC/COAOD will beheld during the 2008 SOFT Confer-ence in Phoenix. The agenda will in-clude the committee’s normal businessand reports from its technical subcom-mittees. The meeting is open to any-one wishing to attend. Please refer toyour program for the time and place.

At the Phoenix meeting thecommittee will announce a recipient tobe awarded the Robert F. BorkensteinAward. This individual will be onewho has a minimum tenure of 25 yearsof active service in the area of alcohol/drugs and traffic safety, has contrib-uted to that field to a degree that theirachievements are nationally recog-nized and has a minimum of 10 yearsof active and productive involvementas a volunteer with the National SafetyCouncil. This year’s awardee is -------?Please keep tuned as many of you longtime SOFT members may want to at-tend that award banquet.

Committee Chair, Dr. SarahKerrigan reports that a DUID websiteis under development. The site willcontain a variety of DUID related re-sources for toxicologists, includingbut not limited to suggested reading,upcoming training events, governmentpublications, DRE and legal informa-tion. A preliminary form of the web-site was presented for discussion pur-poses at the DUID committee meetingin Washington DC, and the committeehopes to have this project completedin October 2008 in time for the SOFTannual meeting in Phoenix.

The SOFT Continuing Educa-tion Committee and the SOFT/AAFSDrugs & Driving Committee recentlyconcluded the “Interpretive DUIDCourse” in concert with the PalmBeach County Sheriff’s Office inWest Palm Beach, Florida on May 6-8th. The class was well attended with25 attendees and six faculty. Sincereappreciation is extended to Tate Yeat-man and Ann Marie Gordon for theirtime and commitment organizing thisworthwhile workshop.

The following is redacted fromthe May/June 2008 “Academy News”by Peter R. Stout, Ph.D., Section Chair:

The submission deadline forAAFS workshops and abstracts is Au-gust 1,2008. Those considering aworkshop should contact Phil Kemp([email protected]. ok.us).

Ken Ferslew is ToxicologySection Program Chair for this year. Ifa member is interested in assisting withthe February 2009 meeting in Denver,Colorado please contact Ken([email protected]).

The Section has formed an adhoc committee consisting of RobertBost, Henry Nipper, John Soper, Chris-topher Boden and Audra Brown thatwill be tasked to evaluate the Section’smembership and work with AAFS tocontact members who may be up forpromotion and provide whatever assis-tance they may need.

The Section elected MarilynHuestis to fill the seat vacated by BarryLogan on the AAFS Board of Direc-tors. William Anderson, Diana Garsideand Loralie Langman were elected toserve on the Awards and ScholarshipCommittee. Nikolas Lemos and Mi-chael Corbett were elected to the Nomi-nating Committee.

T. I . A . F. T. N E W S

La Martinique and the FrenchWest Indies welcomed visitors to the46th annual meeting of the Interna-tional Association of Forensic Toxi-cologists June 2-8, 2008. Two otherorganizations co-hosted this jointmeeting; the French Society of Ana-lytical Toxicology and the Society ofHair Testing.

TIAFT 2009 is scheduled Au-gust 23-27, 2009 in Geneva, Switzer-land. Find details at the TIAFT web-site (www.tiaft.org).

A . A . F. S . F U T U R E

M E E T I N G S I T E S

2009 Denver, CO2010 Seattle, WA2011 Chicago, IL2012 Atlanta, GA2013 Washington, DC2014 Seattle, WA

C . A . T. AT S . O . F. T. I N

P H O E N I X

The California Association ofToxicologists will meet Sunday, Octo-ber 26, 2008 immediately precedingthe S.O.F.T. 2008 meeting in Phoenix.Two half day workshops are sched-uled; “Creating and Giving QualityPresentations” and “Street Drugs &Culture”. Find details at the C.A.T.website (www.cal-tox.org).

Volume 32, Issue 2

TO X IC OL OG Y - B IT S & P I EC ESSubmitted by Section Editor, J. Robert Zettl, MPA

Future S.O.F.T. Meeting Info

2008: Phoenix, AZ……………....Oct. 27-31, 2008…………….Vickie Watts, Norman Wade

2009: Oklahoma City, OK………Oct. 18-23, 2009……………...………………...Phil Kemp

2010: Richmond, VA……………Oct. 18-22, 2010….…..Michelle Peace, Lisa Tarnai Moak

2011: San Francisco, CA………...Aug. 29-Sep. 2, 2011………….………….Nikolas Lemos

2012: Boston, MA……………….June 30-July 6, 2012….……….……...…Michael Wagner

Committee Committee ChairNominating……………………………………Diana Wilkins, Ph.D.Membership………………………. ………….Sarah Kerrigan, Ph.D.Strategic Planning……………………………..Bradford Hepler, Ph.D., DABFTBudget, Finance, and Audit…………………...Robert Turk, Ph.D., DABFTToxTalk Co-Editors…………………………...Yale Caplan, Ph.D., DABFT

Vickie Watts, M.S.ByLaws………………………………………..Yale Caplan, Ph.D., DABFTPublications (JAT Special Issue) ……………..Dan Anderson, M.S., ABFTAwards...………………………………………Philip Kemp, Ph.D., DABFTDrugs & Driving………………………………Sarah Kerrigan, Ph.D.Meeting Resource……………………………..Anthony Costantino, Ph.D., DABFTPolicy and Procedure………………………….William Anderson, Ph.D.SOFT Internet Web-Site………………………Bruce Goldberger, Ph.D., DABFTContinuing Education…………………………Ann Marie Gordon, M.S.Laboratory Guidelines………………………...W. Lee Hearn, Ph,D.Ethics………………………………………….Aaron Jacobs, Ph.D.Drug Facilitated Rape & Sexual Assault……...Marc LeBeau, Ph.D.MS/MS Guidelines……….…………………...John Cody, Ph.D.

February 1 for March Issue

May 1 for June Issue

August 1 for September Issue

November 1 for December Issue

®

The S.O.F.T. 2008 meeting in Phoe-nix will be here before we know it! It takesthe effort of many people to ensure all eventsproceed smoothly. Volunteers are alwaysneeded to help ensure that meeting attendeesfully enjoy their stay in the host city.

Anyone able tooffer a few hours oftime during the meetingas a S.O.F.T. Volunteershould contact DebDenson, the S.O.F.T.Volunteer Coordinatorto schedule times andlocations desirable tohelp with.

Thanks to both past and future volun-teers for helping make S.O.F.T. meetings effi-cient and enjoyable for everyone!

Deb Denson, SOFT Volunteer CoordinatorEmail: [email protected] Tele: 919-541-7265

N E E D V O L U N T E E R S Above: Downtown Phoenix

Below: Papago Buttes, Phoenix

2 0 0 8 S . O . F. T. C O M M I T T E E C H A I R S

S.O.F.T. Administrative OfficeOne Macdonald Center1 N. Macdonald St., Suite 15Mesa, AZ 85201

Phone: 888-866-SOFT (7638)Fax: 480-839-9106E-mail: [email protected]

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Society of ForensicTox icolog ists , Inc.

ToxTalk is the official publication of the Society of Forensic Toxicologists, Inc., mailed quar-terly (bulk mail) to its members. It is each member’s responsibility to report changes ofaddress to the SOFT Administrative Office. Non-members may receive ToxTalk for $15 percalendar year. Checks payable to SOFT may be mailed to the SOFT Administrative Office.To submit articles or address ToxTalk issues please email to [email protected].

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