Arvind K. Chaturvedi

Kristi J. Craft

Doris M. Kupfer

Dennis Burian

Dennis V. Canfield

Civil Aerospace Medical Institute

Federal Aviation Administration

Oklahoma City, OK 73125

October 2009

Final Report

Application of DNA Profiling in

Resolving Aviation Forensic

Toxicology Issues

DOT/FAA/AM-09/19

Office of Aerospace Medicine

Washington, DC 20591

OK-10-0077-JAH

Federal Aviation

Administration
NOTICE

This document is disseminated under the sponsorship 

of the U.S. Department of Transportation in the interest 

of information exchange. The United States Government 

assumes no liability for the contents thereof.

___________

This publication and all Office of Aerospace Medicine 

technical reports are available in full-text from the Civil 

Aerospace Medical Institute’s publications Web site: 

www.faa.gov/library/reports/medical/oamtechreports
i

Technical Report Documentation Page 

1. Report No. 2. Government Accession No. 3. Recipient's Catalog No. 

DOT/FAA/AM-09/19 

4. Title and Subtitle 5. Report Date 

Application of DNA Profiling in Resolving Aviation Forensic October 2009 

Toxicology Issues 6. Performing Organization Code 

 

7. Author(s) 8. Performing Organization Report No. 

Chaturvedi AK, Craft KJ, Kupfer DM, Burian D, Canfield DV 

9. Performing Organization Name and Address 10. Work Unit No. (TRAIS) 

 

11. Contract or Grant No. 

FAA Civil Aerospace Medical Institute 

P.O. Box 25082 

Oklahoma City, OK 73125 

12. Sponsoring Agency name and Address 13. Type of Report and Period Covered 

 

 

 

Office of Aerospace Medicine 

Federal Aviation Administration 

800 Independence Ave., S.W. 

Washington, DC 20591 14. Sponsoring Agency Code 

15. Supplemental Notes 

This work was accomplished under the approved task, AM-B-10-TOX-202. 

16. Abstract 

Biological samples from the victims of aviation accidents are submitted to the Civil Aerospace Medical Institute (CAMI) for 

toxicological evaluation. Body components of aviation accident fatalities are often scattered, disintegrated, commingled, 

contaminated, and/or putrefied at accident scenes. These situations may impose difficulties in victim identification and 

tissue matching, thereby in the toxicological analysis of authentic samples and the interpretation of the associated analytical 

results. The use of DNA typing has been exemplified in the literature to resolve the sample misidentification issue. 

However, the prevalence of this type of issue in relation to aviation accident forensic toxicology has not been wellestablished. Therefore, the CAMI toxicology database was searched for the period of 1998−2008 for those accidents/cases 

wherein DNA profiling was performed. During this period, samples from 3523 accidents were received by CAMI. Of these, 

there were 3366 aviation accidents wherein at least one fatality had occurred. Biological samples from a total of 3319 pilots 

were received. Of these, 3275 were fatally injured. The 3319 pilots translated into the equivalent number of aviation 

accidents. Of the 3319 accidents, there were only 15 (≈ 0.5%) accidents wherein DNA profiling was performed on the 

biological samples. Six occupants (four fatalities and two injured victims) were involved in one accident and five (two 

fatalities and three injured victims) in another. Three fatalities occurred in three accidents each, two fatalities in eight 

accidents each, and one fatality in one accident. In one accident, there were two occupants with non-fatal injuries. DNA 

profiling was conducted upon the requests of families in two accidents, of accident investigators in three, and of pathologists 

in four. In six accidents, contradictory toxicological findings—such as selective presence of analytes in samples—led the 

CAMI laboratory to initiate DNA profiling. The requests made by families and investigators were primarily triggered by the 

inconsistency between the toxicological results and the history of the use of the drugs by the victims, while by the 

pathologists because of the commingling of samples. In three (20%) of the 15 accidents, at least one submitted sample was 

misidentified or mislabeled. The low number of the accident cases requiring DNA profiling suggests that the samplesubmitting agencies take extensive precautionary measures to ensure that the origin of the submitted biological samples are 

correctly identified. Furthermore, the present study confirms that DNA typing can be used as a tool for establishing the 

authenticity of the aviation biosamples, thereby their associated toxicological conclusions.

 

17. Key Words 18. Distribution Statement 

 

 

Forensic Science, Postmortem Toxicology, DNA Typing, 

DNA Profiling, Quality Control/Quality Assurance, 

Aircraft Accident Investigation

Document is available to the public through the 

Defense Technical Information Center, Ft. Belvoir, VA 

22060; and the National Technical Information 

Service, Springfield, VA 22161 

19. Security Classif. (of this report) 20. Security Classif. (of this page) 21. No. of Pages 22. Price 

Unclassified Unclassified 15 

Form DOT F 1700.7 (8-72) Reproduction of completed page authorized 

iii

Contents

INTRODUCTION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

Biological Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

Analytical Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Toxicology Database. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

DNA Profiling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

CASE HISTORIES. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

RESULTS AND DISCUSSION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

REFERENCES. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

1

Application of DNA Profiling in Resolving

Aviation Forensic Toxicology Issues

INTRODUCTION

During aviation accident investigations, biological

samples are collectedfromvictimsbylocalpathologists and

submittedtotheFederalAviationAdministration’s(FAA’s)

Civil Aerospace Medical Institute (CAMI; Oklahoma

City,OK)fortoxicologicalevaluation.

1,2 Such shipments

includesamplesfrompilotfatalities,survivedpilots,other

crewmembers, andpassengers. Thesubmissionofsamples

from passengers, however, depends upon the nature of

an accident, such as an accident involving fire.

While maintaining a high degree of quality assurance

andcontrol,3-11 acquiringaccurate andauthentic analytical

dataonthebiologicalevidencehasbeentheprimaryobjective of CAMI in order to effectively assist the conclusion

of aircraft accident investigations. However, the accuracy

and authenticity of such collected data fundamentally

depends upon the integrity of the submitted biological

samples, thereby allowing the truthful interpretation

of results with a high degree of confidence and judicial

admissibility.

There is a potential for inherited realistic limitations

in aircraft accidents, particularly in those wherein more

than onefatality had occurred and, additionally, wherein

multiple types of postmortem specimens were collected

and submitted. In some instances, samples originating

from an incorrect source would have been submitted or

they were labeled with an incorrect name—that is, the

sample(s) are misidentified or mislabeled, possibly due

to human error. Such potential is because of theintrinsic

nature of aviation accidents wherein body parts of fatally

injured victims are often scattered, disintegrated, commingled, contaminated, and/or putrefied. Of course,

the extent of such accident scene conditions primarily

depends upon the severity and situation of a particular

accident. These circumstances may impose difficulties

for victim identification, tissue matching, and thereby

authentic sample analysis andresultinterpretation. However, these issues, at least to some extent, can be resolved

by DNA profiling.

The use of DNA profiling has previously been documented in resolving tissue mismatching and/or analytical result interpretation issues for two aviation accident

cases.

12 One case was associated with the uncertainty of

the correct identity of the tissues originated from either

and/or both of the two occupants, while the other case

with the selective presence of atropine. Although the

application of DNA profiling has been exemplified, the

prevalence of sample identity/validity and result interpretation issues in association with aviation accident

forensic toxicology has not been established. Therefore,

the CAMI toxicology database was searched for those

aviation accident cases wherein DNA profiling was

performed. The search period was from 1998 to 2008. 

In the present study, those DNA-related accidents/cases

are described, covering the associated histories, findings,

discussions, and conclusions.

MATERIALS AND METHODS

Biological Samples

Biological specimens collected from aviation accident casualties are submitted to CAMI in the FAA

TOX-BOX evidence containers at the request of the

National Transportation Safety Board (NTSB).

1,2 It is

strongly recommended that one TOX-BOX evidence

container per victim be used for shipping samples. The

types of samples generally received at CAMI are blood,

urine, vitreous fluid, spinal fluid, brain, lung, heart, liver,

kidney, muscle, and/or other body tissues. Reference

material—for example, biological samples from blood

relatives and personal effects of victims—may also be

received for DNA profiling.

The TOX-BOX evidence containers are received in

the secured accessioning area of the CAMI’s Bioaeronautical Sciences Research Laboratory. Upon receipt,

the containers are opened in the presence of two (or

more) authorized quality assurance/quality control staff

members. Only one evidence container is processed at

a time; other containers are left intact. The processing

involves the opening of the container and cataloging of

its contents, which entails videotaping the outside and

inside of the container, including seals and labels, and

documenting the details of the case, including chain-ofcustody. This approach istaken according to thestandard

operating procedure of thelaboratory. After processing a

case in one TOX-BOX evidence container, the evidence

container of another caseisstarted. All videotapes arelater

reviewed to ensure the chain of custody of the container

and its forensic contents. The policy of “one-container

(case)-at-a-time” is always followed. This philosophy is

alsopracticedduring the aliquotingofsamplesofthe cases
2

for the preparation of batches for analyses. Each victim

of an accident from which samples are received is given

a specific CAMI case number. Therefore, the “accident,”

“victim,” and “case” words are considered related to each

other and are interchangeably used in the present study,

when necessary.

Analytical Toxicology

The presence of combustion gases, ethanol/volatiles,

and drugs in the samples is analytically demonstrated

by screening, followed by confirmation and/or quantitation.

2,13,14 All analyses are performed according to establishedstandardoperatingproceduresofthelaboratory

by using various techniques such as spectrophotometry,

immunoassay, and chromatography. The combustion

gasesentail carbon monoxide and hydrogen cyanide; the

drugs include a wide range of prescription, nonprescription, and illegal drugs. DNA profiling is performed on

case samples in which there is doubt about the identity

of the submitted samples. This DNA approach becomes

a necessity when there are reasons to believe that the

samples might have been misidentified (mislabeled),

mismatched, or commingled with the samples of other

victims during the collection of samples. These toxicological and DNA profiling aspects are summarized in a

recent review article.

15

Toxicology Database

Analytical toxicology results, including DNA typing

findings, areelectronically enteredinadatabase atCAMI,

also incorporating relevant information of the associated

accidents and victims. In thisCAMItoxicology database,

the detailed DNA profiling findings of aviation accident

cases have been stored since 1998. For the present study,

the toxicology database was examined for the accidents

andforthefatally/non-fatally injuredpilots andother victimswhereinDNAtyping analyseswere performed. This

evaluation included the associated toxicological findings,

as well. The selected period for the examination was 11

years(1998–2008). Forthis period,the databasewas also

searched for the total number of all accidents, including

aviation accidents and associated pilot fatalities whose

postmortem biological samples were received at CAMI.

DNA Profiling

DNA profiling on the identity-related samples and/

or associated reference samples (or evidence) were performed by the FAA’s CAMI laboratory or by an outside

laboratory. The selection of the laboratory for the profiling was governed by the accident scenario and/or by the

aviation accident investigator’s decision. The number

and type of DNA loci evaluated were dependent upon

the standard operating procedures of the laboratories

from which DNA analytical services were obtained and

the state-of-the-art techniques available at the time of

DNA profiling. For example, DQA1 and poly-markers

(PMs)—LDLR, GYPA, HBGG, D7S8, and GC—were

evaluated during the 1990s.

12,16-18 However, as advancements in the technology took place during the 2000s,

the number of DNA loci included in the protocols of

the laboratories increased from half a dozen up to more

than a dozen (D3S1358,D16S539,D8S1179, D21S11,

D18S51, D5S818, D13S317, D7S820, CSF1PO,

TPOX, TH01, vWA, FGA, Penta E, Penta D, SRY-ZF,

and amelogenin19-24). In some situations, an evaluation

of a gender marker—for example, amelogenin23 or SRYZF19,21—sufficed the need to deduce the origin of the

tissues. This approachwasparticularlymorerealisticwhen

there were only two occupants involved in an accident

and they were of the opposite sex.

Theprocedure adoptedintheCAMIlaboratoryduring

the 1990sforDQA1 and PMs has been reported earlier.

12

Briefly,the procedure consisted oftheextraction ofDNA

from biological samples, followed by DNA quantitation

and amplification, and polymerase chain reaction (PCR)

product verification, hybridization, and detection.

16-18

During the 2000s, several new genetic markers were

incorporated in the protocol of the CAMI laboratory.

25

In general, DNA profiling on samples was performed by

comparing PCR amplicon lengths generated from nine

autosomal loci and one or two gender determination loci

by electrophoretic mobility.

25 The short tandem repeat

(STR) loci are part of the Federal Bureau of Investigation’sCombinedDNAIndex System—thatis,CODIS.

26

The commonly used loci in CAMI’s laboratory were

D3S1358, D5S818, D7S820, D13S317, D16S539,

FGA, THO1, TPOX, and vWA. These STR loci are

variable four-nucleotide tandem repeats. Reactions

for each autosomal locus were individually performed. 

There was no multiplexing. For gender determination,

amelogenin23wasinitiallyused. Subsequently,thismarker

was replaced with a duplex reaction for establishing the

presence of the Y-chromosome-specific SRY locus and

the homologous zinc finger protein genes, ZFX/ZFY, of

theXandY chromosomes.

27 PCRamplicon lengthswere

compared by micro-fluidic electrophoresis on an Agilent

BioAnalyzer 2100 (SantaClara,CA) by usingDNA1000

series-2 chips that were processed following the Agilent’s

DNA1000 series-2 protocol for this instrument. At the

expected fragment lengths for these loci, the instrument

has four-nucleotide resolution.
3

CASE HISTORIES

Case 1

Scenario

An experimental aircraft crashed during a personal

pleasure flight, killing both occupants—the pilot and

the passenger. This flight was being conducted to demonstrate the performance capabilities of the aircraft to

the passenger. The aircraft was destroyed. This accident

occurred in 1998. Autopsies on both victims were performed, and postmortem samples from the victims were

submitted to CAMI.

Submitted Samples

Pilot: Bile, brain, gastric, heart, kidney, liver, lung,

muscle, spleen, and urine

Passenger: Bile, brain, gastric, heart, kidney, liver,

lung, muscle, and spleen

Toxicological Findings

Pilot: Negative

Passenger: Diphenhydramine 0.075 μg/mL in liver

fluid and 0.08 μg/mL in kidney fluid

Reason for DNA Profiling

Upon the request of the passenger’s family, DNA

typing of the passenger’s “so called” brain sample was

performed by an external laboratory in relation to the

reference samples—hair from the passenger and blood

from the passenger’s daughter. The DNA typing of the

hair and blood samples did not exclude the passenger

as the biological father of the daughter, but the DNA

profile of the brain suggested that it did not originate

from the passenger.

The abovesample ambiguity led theCAMIlaboratory

toperformadditionalDNAanalysisonallsubmittedsolid

tissuesamplesfromthepilot, aswell asfromthepassenger. 

Exceptforbrainsamples, allothersampletypesoriginated

from two different sources. The brain samples labeled as

“pilot” andas “passenger”matchedwitheachother. These

findings, in conjunctionwith theearlierreferencesample

analysis, concluded that both submitted brain samples

originated from the same individual—that is, the pilot,

not the passenger. Therefore, it was concluded that all

samples, except brain samples, were correctly labeled at

thetime of autopsy as “pilot” and as “passenger,” and the

toxicologicalfindingsofthepresenceofdiphenhydramine

in liver and kidney were correctly associated with the

passenger, not with the pilot. No toxicological analysis

was performed on the brain samples.

DNA Loci Evaluated

The passenger’s hair, daughter’s blood, and brain

samples were evaluated for D2S44, D18S27, D4S163,

D7S21, DQA1, LDLR, GYPA, HBGG, D7S8, and GC

by an external laboratory; and all solid samples from the

pilot andthepassengerforDQA1,LDLR,GYPA,HBGG,

D7S8, and GC by the CAMI laboratory.

Case 2

Scenario

In 1998, a two-occupant aircraft on a maintenance

test flight crashed, killing the pilot and the copilot. The

aircraft was destroyed. At the time of accident, visual

meteorological conditions prevailed. A muscle sample

collected from the pilot was submitted to CAMI.

Submitted Samples

Pilot: Muscle

Copilot: No sample

Toxicological Findings

Pilot: Negative

Copilot: Not applicable

Reason for DNA Profiling

CAMIwasrequested to conduct DNA analysis on the

musclesample, sincethe pathologist was unsure whether

themuscleremains belonged to the pilot orto the copilot. 

Reference items (hair dryer, hair brush, toothbrush, and

two dental picks from the pilot;electric razor and toothbrushfromthe copilot)werelatersubmittedtoCAMIfor

theDNAexamination. TheDNAprofiling ofthemuscle

sample and of the hair sample taken from the hair dryer

labeled as “pilot” suggested the genotype of thesesamples

werethesame. DNA found on thetoothbrush labeled as

“copilot” did not match with that of the muscle. Based

upon the CAMI laboratory analysis, it was concluded

that the source of the muscle was indeed the pilot.

DNA Loci Evaluated

The DNA lociexaminedwere DQA1, LDLR, GYPA,

HBGG, D7S8, and GC.

Case 3

Scenario

Afloat-equippedairplanesustainedsubstantialdamage

when it collided with the ground. There were two occupants in the plane, the first pilot/flight instructor and the

student pilot. Both occupants received serious injuries in

this2000non-fatal accident. Thefirstpilot/flightinstructor was a male, and the student pilot was his daughter. 

The accident-investigator-in-charge coordinatedwiththe

local authorities the shipment of the biological samples
4

collected from the male pilot to CAMI. Samples from

thefemale pilotwere notsubmitted, asitwas determined

that the aircraft was flown by the male pilot.

Submitted Samples

Pilot: Blood, serum, and urine

Student Pilot: None

Toxicological Findings

Diphenhydramine 0.018 μg/mL in blood

Bupropion metabolite present in blood

Acetaminophen 14.3 μg/mL in urine

Diphenhydramine, bupropion, and lidocaine present

in urine

Reason for DNA Profiling

Baseduponthetoxicologicalfindings,itwasviewedthat

the pilotwas flyingwhile using unapprovedmedications. 

Themalepilot claimedthatthe analyzedsamplesbelonged

to his daughter and not to him; accordingly, he declared

that it was his daughter who was on the medications and

not him. Thus, the wrong specimens were analyzed. The

NTSB accident investigator requested DNA analysis to

determinethe gender and origin ofthesource. The blood

sample was sent to an outside laboratory for the DNA

analysis. The amelogenin locus was used for the gender

identificationofthe originofthe blood sample and found

that the sample was from a single male human source. 

Therefore,itwas concludedthatthetoxicologically tested

samples originated from a male (the pilot), not from a

female (the student pilot—daughter).

DNA Locus Evaluated

The locus was amelogenin.

Case 4

Scenario

In 2000, an aircraft collided with the terrain, fatally

injuring the pilot and two passengers. The airplane was

destroyed. Bodies of all the three victims were severely

fragmented and commingled. Several pieces of tissue

samples were shipped to CAMI in one TOX-BOX evidence container.

Submitted Samples

Pilot/Passengers: Body tissue, kidney, lung, and

muscle

Toxicological Findings

Pilot: Negative

Passengers: No analysis

Reason for DNA Profiling

The NTSB accident investigator requested DNA

profiling because samples from the three fatalities were

severely fragmented. The DNA profiling was conducted

by an external laboratory. Based upon the DNA profiling findings and the accident scene sector numbers, the

samples were labeled with the respective sector numbers

and were accordingly identified as to befrom pilot (body

tissue, kidney, lung, and muscle), passenger I (body

tissue), and passenger II (body tissue and muscle). All

samples were submitted to CAMI in one TOX-BOX

evidence container. Knowing the source of the samples,

the toxicological analysis was performed on the samples

originated from the pilot. Samples from passengers were

not toxicologically evaluated.

DNA Loci Evaluated

The loci were D3S1358, VWA, FGA, D8S1179,

D21S11, D18S51, D5S818, D13S317, D7S820,

CSF1PO, TPOX, TH01, D16S539, and amelogenin.

Case 5

Scenario

In 2001, a cargo planeimpacted theterrain following

an uncontrolled descent and was totally destroyed. One

of thetwo occupants of the plane died at thescene of the

accident. Thesecond occupant was seriously injured but

diedlater. PostmortemsamplesweresubmittedtoCAMI.

Submitted Samples

Pilot/Copilot: Three pieces of muscle

Toxicological Findings

Pilot: Negative

Copilot: Negative

Reason for DNA Profiling

The three pieces of muscle were submitted in one

TOX-BOX evidence container. The samples were commingled, and the origin of the muscles could not be

positively established by the pathologist. Because of the

source uncertainty, samples from the muscles were subjected to DNA profiling. The profiling was performed

at an external laboratory. Based upon the DNA results,

the muscles were separated in two groups—one muscle

piece belonged to one person and the remaining two

pieces to the second person. However, in the absence

of any reference sample, it was not possible to link the

origin of those groups specifically to the pilot or copilot. 

Since toxicological findings were negative in both cases,

the origin association was not relevant.
5

DNA Loci Evaluated

The loci were D3S1358, TH01, D21S11, D18S51,

Penta E, D5S818, D13S317, D7S820, D16S539,

CSF1PO,PentaD, amelogenin, vWA,D8S1179,TPOX,

and FGA.

Case 6

Scenario

An aircraft piloted by a private pilot was destroyed

on impact in a pasture. This flight was associated with a

pheasanthunting expedition. Inthis 2004 accident,there

were three occupants (the pilot and two passengers), all

of whom werefatally injured. Body parts of victims were

badly fragmented and were also commingled among

approximately 27 kg of pheasant. Although there were

three fatalities, biological samples, presumably from the

pilot, were submitted to CAMI.

Submitted Samples

Pilot: Bone, muscle, skin, and lung

Toxicological Findings

Pilot: Propoxyphene 0.308 μg/g in muscle and 0.341

μg/g in skin

Norpropoxyphene 0.671 μg/g in muscle and 0.123

μg/g in skin

Bupropion present in muscle

Dextromethorphan and tramadol present in muscle

and skin

Passenger (Post-DNA Analysis): Pseudoephedrine

present in lung

Reason for DNA Profiling

The pathologist requested that CAMI perform DNA

profiling on the submitted samples to ensure that they

truly originated from one human being—that is, the

pilot. Muscle,skin, and lung specimensweresubjected to

the DNA analysis by the CAMI laboratory. The analysis

revealed that the muscle and the skin came from one

victim, whilethelung from a second victim. These DNA

findings warranted creating a new case for the second

victim—that is, one of thetwo passengers—and thelung

sample was transferred to that case. Thesetwo groupings

were further supported by the toxicological findings. 

The drugs found in the muscle and skin specimens were

consistent with each other, whereas the drug in the lung

was not,supporting the DNA resultsthat the muscle and

skin came from one individual, while the lung from a

different individual.

DNA Loci Evaluated

The samples were examined for the following loci:

amelogenin, CSF1PO, D5S818, D7S820, D13S317,

D16S539, TH01, TPOX, and vWA.

Case 7

Scenario

A single-engine aircraft was destroyed during impact

with terrain following a loss of control during an approach. The two occupants—the flight instructor and

the student pilot—did not survive the crash. There was

no fire. Postmortem samples from both victims were

submitted to CAMI.

Submitted Samples

Flight Instructor: Bile, blood, brain, gastric, heart,

kidney, liver, lung, muscle, spleen, and urine

Student Pilot: Bile,blood,brain,gastric,heart,kidney,

liver, lung, muscle, spleen, urine, and vitreous fluid

Toxicological Findings

Flight Instructor: Atenololdetectedinbloodandurine

Student Pilot: Cyanide 3.7 μg/mL in blood

Reason for DNA Profiling

There was no firein this accident. The blood from the

instructor was negative for cyanide, but the blood from

the student was found to be positive for cyanide. The

bloodcyanide concentrationwasinthelethalrange. Such

selective presence of cyanideled the CAMI laboratory to

perform DNA profiling on the cyanide-positive blood

sample, along with the associated sample types used for

othertoxicology tests. These othersampletypeswereliver

and kidney. The DNA analysis disclosed that the blood,

liver, andkidneywerefromthesameindividual. Although

the DNA analysis was able to ensure that these three

samples were of one origin, the reason for the selective

presence of cyanide in the high concentration could not

be toxicologically deduced, particularly when there was

no fire and the carboxyhemoglobin level was negative.

DNA Loci Evaluated

The loci used for the DNA typing were amelogenin,

D3S1358, D5S818, D7S820, D13S317, D16S539,

TH01, TPOX, and vWA.

Case 8

Scenario

In 2006, an aircraft was destroyed when it impacted

water and lightstanchionswhile approaching the airport. 

There were two pilots and three passengers. The pilots
6

were fatally injured, but the passengers received only

minorinjuries. Postmortemsamplesfromthe pilotswere

submitted to CAMI.

Submitted Samples

Pilot: Bile, blood, brain, gastric, heart, kidney, liver,

lung, muscle, spleen, urine, and vitreous fluid

Copilot: Bile, blood, brain, gastric, heart, kidney,

liver, lung, muscle, spleen, urine, and vitreous fluid

Toxicological Findings

Pilot: Cocaine 0.202 μg/mL in urine

Benzoylecgonine 0.445 μg/mL in urine

Ecgonine methyl ester and quinine present in urine

Copilot: Ephedrine, pseudoephedrine, and phenylpropanolamine present in urine

Reason for DNA Profiling

DNA analysis was performed because cocaine was

found in urine but not in the pilot’s blood. It was suspected that the blood might have originated from the

copilot. To confirm the suspicion, the DNA typing was

performed by theCAMIlaboratory on the blood samples

from both victims. The DNA profiling results concluded

that the submitted blood samples originated from two

different sources. Thus, the blood that was analyzed for

cocaine was truly negative for cocaine.

DNA Loci Evaluated

The DNA loci entailed SRY-ZF, D3S1358, D5S818,

D7S820,D13S317,D16S539,TH01,TPOX, andvWA.

Case 9

Scenario

An aircraft collided with terrain during a circling instrument approachandwasdestroyed. This accidenttook

placein 2007. Both the pilot and a passenger were killed

andtheirbodieswereextensivelyfragmented. Postmortem

samples from both victims were submitted to CAMI.

Submitted Samples

Pilot: Blood, lung, muscle, and spleen

Passenger: Blood, gastric, kidney, liver, lung, muscle,

and spleen

Toxicological Findings

Pilot: Negative

Passenger: Negative

Reason for DNA Profiling

The pathologist informed CAMI that organs of the

aviation accident victimswere commingled. Adoubtwas

raised whether the submitted blood samples collected

from a disaster pouch originated from a singleindividual

or if they were mixtures of blood originating from both

individuals. CAMI performed DNA profiling on Blood

1, Blood 2, and muscle samples from the pilot (Group

1) and on Blood 1, muscle, and liver samples from the

passenger (Group 2). The DNA analysis of the three

sample types from the pilot (Group 1) confirmed that

they originatedfromoneindividual. Themuscle andliver

of Group 2 belonged to a different individual, which was

deduced to be of the passenger. The “so called” Blood 1

samplefromthepassengerwasdeterminedtobe amixture

of two different genetic origins—that is, from the pilot

and the passenger. Therefore, Blood 1 was not used for

toxicological evaluation.

DNA Loci Evaluated

TheDNAprofilingwasestablishedbyusingthefollowing markers: amelogenin, D3S1358, D5S818, D7S820,

D13S317, D16S539, TH01, TPOX, and vWA.

Case 10

Scenario

Following a descent from cruising altitude, an aircraft with two occupants was destroyed on impact with

terrain. Both occupants—the pilot and the pilot-rated

passenger—sustained fatal injuries in this 2007 crash. 

Postmortem samples were submitted to CAMI.

Submitted Samples

Pilot: Lung and muscle

Passenger: Kidney and liver

Toxicological Findings

Pilot: Ethanol 43 mg/hg in muscle and 59 mg/hg

in lung

Atenolol present in lung and muscle

Passenger: Ethanol 26 mg/hg in liver and 25 mg/

hg in kidney

Atenolol present in liver and kidney

Reason for DNA Profiling

Lung and muscle samples were submitted together in

onesingle plastic bag. Upon receiving the bag, thesetwo

tissuetypeswereseparated and placed individually in two

different bags. Similarly, liver and kidney samples from

the second victim were submitted in one single plastic

bag; thesesampletypes wereseparated and placed in two

different bags. Because of thesamplesubmissions of two

sample types in one bag and the presence of ethanol and

atenolol in samples of both victims, there was a suspicion

whether the submitted samples originated from one individual. The four samples—lung and muscle from the

pilot and liver and kidney from thesecond victim—were
7

subjected toDNAprofiling at theCAMIlaboratory. The

DNA results suggested that lung and muscle originated

from one male subject, while liver and kidney from a

female subject. The presence of ethanol and atenolol in

the sample types from both subjects was determined to

be coincidental.

DNA Loci Evaluated

The DNA loci were SRY-ZF, D3S1358, D5S818,

D7S820,D13S317,D16S539,TH01,TPOX, andvWA.

Case 11

Scenario

This 2007 accident involved an aircraft that was

destroyed upon colliding with terrain after takeoff. The

sole occupant of the aircraft did not survive the crash. 

Postmortem samples from the victim were submitted

to CAMI.

Submitted Samples

Pilot: Blood, brain, gastric, heart, kidney, liver, lung,

muscle, spleen, and vitreous fluid

Toxicological Findings

Pilot: Negative

Reason for DNA Profiling

An external laboratory detected hydrocodone in the

victim’s blood, but the drug was absent in urine. CAMI’s

toxicologicalevaluation failed to disclosethe presence of

any drugs/alcohol and,therefore,the casewas considered

as negative. The discrepancy in the toxicological findings—positive versusnegative—fromthetwolaboratories,

led the CAMI laboratory to subject the blood, kidney,

and muscle samples for DNA profiling. These samples

were found to be from the same male. This conclusion

was consistent with the fact that the pilot was the sole

male occupant of the crashed aircraft. It could thus be

deduced that the blood sample analyzed by the outside

laboratory might have originated from a person other

than the pilot. The analyzed blood was possibly misidentified as from “pilot,” leading to the reporting of a

“false” positive for hydrocodone in the pilot’s blood by

the external laboratory.

DNA Loci Evaluated

Thelocievaluated were SRY-ZF, D3S1358, D5S818,

D7S820,D13S317,D16S539,TH01,TPOX, andvWA.

Case 12

Scenario

Anaircraftwassubstantiallydamagedwhenitimpacted

the ground while on approach to the airport. Both occupants—the pilot and the passenger—sustained fatal

injuries. Postmortembiologicalsamplesfromthe victims

were submitted to CAMI. 

Submitted Samples

Pilot: Blood, brain, gastric, heart, liver, lung, muscle,

and spleen

Passenger: Blood, brain, heart, kidney, liver, lung,

muscle, spleen, and urine

Toxicological Findings

Pilot: Atenolol present in liver and lung

Passenger: Atenolol present in liver and urine

Reason for DNA Profiling

Both victims’ toxicology suggested the presence of

atenolol in their systems and their medical records

indicated that they were not on any medication, so it

was decided to perform DNA typing to ensure that the

samples did not originatefrom the sameindividual. The

DNA profiling was performed at the CAMI laboratory. 

The samples were liver and lung from the pilot (Group

1), and also liver and lung from the passenger (Group 2). 

The DNA profiling findings concluded that the Group

1 samples originated from one individual, while the

Group 2 samples from another individual. Therefore,

the presence of atenolol in the systems of both victims

was coincidental.

DNA Loci Evaluated

The DNA loci used for the typing were SRY-ZF,

D3S1358, D5S818, D7S820, D13S317, D16S539,

TH01, TPOX, and vWA.

Case 13

Scenario

An amateur-built aircraft was destroyed when it

impacted trees and terrain after missing an approach to

landing. Boththepilot andthe copilotwerefatally injured. 

Postmortemsamplesfromboth victimswere collected by

the pathologist and submitted to the CAMI laboratory. 

This aviation accident occurred in 2008.

Submitted Samples

Pilot: Kidney, liver, and muscle

Passenger: Kidney, liver, and muscle
8

Toxicological Findings (Pre-DNA Analysis)

Pilot: Ethanol 15 mg/hg in liver

Ethanol not present in muscle

Amlodipine present in liver and kidney

Losartan and diphenhydramine present in liver

Copilot: Ethanol 24 mg/hg in muscle

Toxicological Findings (Post-DNA Analysis)

Pilot: Negative

Copilot: Ethanol 15 mg/hg in liver and 24 mg/hg

in muscle

Amlodipine present in liver and kidney

Losartan and diphenhydramine present in liver

Reason for DNA Profiling

Uponreceiving thetoxicologicalreport, anuncertainty

was raised by the family members of the pilot that there

was something wrong in the sample collection and/or

toxicological evaluation processes. To the best of the

family members’ knowledge, the pilot was not on the

medications mentioned in the toxicology report. The

NTSB suspected that there was a sample mix-up at

the time of autopsy and asked for DNA analysis to be

performed by the CAMI laboratory. It appeared that the

samples were mislabeled. A reference DNA sample from

thepilot’ssonwas collectedasbuccalswabs andsubmitted

to CAMI. With reference to the son’s DNA typing, the

submitted liver, kidney, and muscle samples labeled as

“pilot” and the muscle sample labeled as “copilot” were

subjected to DNA typing. The DNA findings concluded

that the liver and kidney samples originally labeled as

“pilot” originated from the copilot, instead of the pilot,

but the submitted muscles were correctly labeled as to

their respective origins. Thus, both toxicological reports

were accordingly corrected (see the post-DNA analysis

toxicological findings mentioned earlier). The supplemental report findings were consistent with the medical

histories of the pilot and of the copilot. In the copilot’s

medical certification records, the use of amlodipine and

losartan was documented. In addition to the modifications in the reports, the originally submitted samples

were correspondingly labeled correctly and transferred

to the respective case storage bins in the CAMI laboratory. Necessary changes in the folders of both cases were

also made and documented to rectify the mislabeling of

specimens during autopsy.

DNA Loci Evaluated

The DNA loci examined were SRY-ZF, D3S1358,

D5S818, D7S820, D13S317, D16S539, FGA, TH01,

TPOX, and vWA.

Case 14

Scenario

In an aviation accident that occurred in 2008, three

occupants—the pilot and two passengers—were fatally

injured. One of the occupants was female. This accident

happened when the aircraft impacted water following a

loss of control. The wreckage cameto rest approximately

12 m below the surface of the water. Under the water,

bodies of the victims were completely fragmented, and

their parts were scattered and commingled. Because of

the water depth, it was difficult for the investigators to

properly retrieve the remains of the victims and to correctly establish the identity of the recovered body parts. 

Bodypartsdidnothave anatomically identifying features. 

Considering the complexity of the accident, the recovered samples were packed in three TOX-BOX evidence

containers and shipped to the CAMI laboratory. Each

of the three containers was intended to contain samples

originating from one of the three victims, but the potential for the presence of the body parts from the other

victims in any given evidence container was very strong. 

Therefore, the misidentification of the submitted body

tissues and their commingling could not be ruled out.

Submitted Samples

Pilot/Passengers: Large pieces of body parts

Toxicological Findings

Pilot: Negative

Female Passenger: Negative

Male Passenger: Diazepam, nordiazepam, and zolpidem present in body tissue

Reason for DNA Profiling

Because ofthe complexity ofthe case and the potential

of the misidentification of the body parts, the accident

investigator requested that DNA profiling be performed

by theCAMIlaboratory. Referencesamples—hairbrush,

toothbrush, razor, and comb belonging to each victim—

were provided for the DNA analysis. Buccal swabs from

the female victim’s son were also submitted.

To ensure sufficient amounts of samples available for

the postmortemtoxicology,threelarge pieces ofthe body

parts were taken out from each evidence container and

wereproperlymarkedwithuniqueidentificationnumbers. 

Small portions of these nine body parts were subjected

to the DNA analysis, along with the respective reference

samples andthebuccalswab. BasedupontheDNAtyping

findings,the nine body partswereseparated and grouped

into three. Such separation and groupingwas based upon

theDNAprofilematching ofthe ninetissuesampleswith

that of the reference samples and buccal swab, including
9

theDNA-basedgenderidentification. Thesethree groups

wereidentified as the pilot, thefemale passenger, and the

male passenger. After this DNA-based identification and

grouping, thesamples from thethree victims—thethree

groups of body tissues—were toxicologically evaluated. 

Thetoxicologicalfindings aresummarizedintheprevious

subsection of this case.

DNA Loci Evaluated

The loci were SRY-ZF, D3S1358, D5S818, D7S820,

D13S317, D16S539, FGA, TH01, TPOX, and vWA.

Case 15

Scenario

In a 2008 accident, an aircraft overran the runway

while taking off and crashed. Tire debris and portions of

airplane components were found along the runway. The

beginning of the takeoff roll appeared normal and then

sparks were observed. The crew attempted to reject the

takeoff, but was unable to stop the airplane. The plane

continued beyond the runway and crashed through airport lighting, navigation facilities, fence, and a roadway,

and came to stop. There was a postcrash fire. The two

crewmembers and two of thefour passengers werefatally

injured. The remaining passengers suffered serious injuries. Postmortem samples from the crewmembers—the

pilot and the copilot—were submitted to CAMI.

Submitted Samples

Pilot: Bile, blood, brain, gastric, heart, kidney, liver,

lung, muscle, spleen, and urine

Copilot: Bile,blood,brain, gastric,heart,kidney,liver,

lung, muscle, spleen, and vitreous fluid

Toxicological Findings

Pilot: Carboxyhemoglobin 20%

Cyanide 1.80 μg/mL in blood

Diphenhydramine 0.030 μg/mL in blood

Diphenhydramine present in liver

Copilot: Carboxyhemoglobin 25%

Cyanide 2.07 μg/mL in blood

Diphenhydramine 0.036 μg/mL in blood

Diphenhydramine present in liver and urine

Ibuprofen present in urine

Reason for DNA Profiling

Becausethetoxicological results weresimilar for both

victims, DNA profiling was performed by the CAMI

laboratory. Thesamplestestedfortheprofilingwereblood

and liver from the pilot, as well as from the copilot. The

results revealed that all specimens tested were indeed

from the respective female (pilot) and male (copilot)

victims, as indicated by the specimen labels provided by

the pathologist.

DNA Loci Evaluated

The DNA loci were SRY-ZF, D3S1358, D5S818,

D7S820, D13S317, D16S539, FGA, TH01, TPOX,

and vWA.

RESULTS AND DISCUSSION

During the 11-year period (1998–2008), biological

samples from the casualties of a total of 3523 accidents

were submitted to CAMI for toxicological evaluation. 

Of these, 3366 were fatal aviation accidents—that is, at

least onefatality had occurred in each of these accidents. 

Biological samples from a total of 3319 pilots were submitted; of these pilots, 3275 were fatally injured. The

3319 pilots translated into the same number of aviation

accidents. Out of the 3319 accidents, there were only15

(≈0.5%) accidents—onenon-fatal and14fatal—wherein

DNA profiling was performed to resolve the issue of the

identity of thesubmitted samples. In someinstances, additionalevidence—suchasbiologicalsamplesfromblood

relatives and tooth brush and hairs of the victims—was

included in the DNA typing to resolvetheidentity issue.

In these 15 accidents, the number of DNA loci

evaluated ranged from one to 16. Depending upon the

nature of an aviation accident and the number of occupants, the DNA loci examined had sufficient power of

discrimination to resolve the origin issue of the accident

case samples. For example, if there were two occupants

of opposite sex in an accident, then one DNA locus of a

genetic marker for the determination of gender sufficed

the need for establishing the origin of the samples. In

other situations wherein occupants were of thesamesex,

the higher number (six to 16) of DNA loci was desirable for establishing the source of the samples with an

acceptable, high degree of certainty. This acceptability

was decisive because the number of occupants in any

given accident was small (one to six) and well-defined

by DNA analysis. This approach was within the realm

of the application of DNA typing for a small number of

individuals in a population.

Of the 15 accidents, six occupants (four fatalities and

twoinjuredvictims)wereinvolvedinone accident andfive

(two fatalities andthreeinjuredvictims)inanother. Three

fatalities occurred in three accidents each, two fatalities

in eight accidents each, and one fatality in one accident. 

In one accident, thereweretwo occupantswith non-fatal

injuries. The DNA profiling of the collected biological

specimens from these 15 accidents was conducted upon

the requests of families in two accidents, of accident
10

investigators in three, and of pathologists in four. In six

accidents, contradictory toxicological findings—such as

selective presence of analytes in samples or presence of

same analytes in the samples from each occupant—led

the CAMI laboratory to initiate the DNA profiling to

ensuretheidentityofthesubmittedsamplesofthe victims. 

The requests made by families and investigators were

also primarily triggered by theinconsistency between the

toxicologicalresults andthe aeromedicalhistoryoftheuse

of the drugs by the victims, while the pathologists were

concerned because of the commingling of samples. In

three (20%) of the 15 accidents, at least, one submitted

sample was determined to be misidentified or wrongly

labeled, presumably dueto human error during autopsy.

Because of the fundamental nature of the seriousness

of aviation accidents consisting of commingling, scattering, disintegration, and putrefaction of bodies and

their parts at the accident scenes, the potential for the

samples to be misidentified with respect to their origin

is a real challenge for the accident investigators and pathologists. In spite of these limitations, the findings of

this study suggests that the sample submitting agencies

take extensive precautionary measures to make certain

that the origin of the submitted biological samples are

correctlyidentified,thoughthepotentialformisidentification/mislabeling of samples still exits. This conclusion is

supported by the fact that there were only ≈ 0.5% of the

accidents wherein the DNA profiling was requested and

the mislabeling of samples was found in only two (13%)

of the 15 accidents. The present study confirms that the

DNA typing could be used12 as a tool forestablishing the

authenticity of the submitted samples, and thereby their

associatedtoxicological conclusions. Basedonthefindings

of the present study, it is concluded that mislabeling of

specimens appears to occur rarely.

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