Arvind K. Chaturvedi 

Patrick S. Cardona

John W. Soper

Dennis V. Canfield

Civil Aerospace Medical Institute

Federal Aviation Administration

Oklahoma City, Oklahoma 73125

December 2003

Final Report

This document is available to the public

through the National Technical Information

Service, Springfield, Virginia 22161.

 

Office of Aerospace Medicine

Washington, DC 20591

DOT/FAA/AM-03/22 Distribution and Optical Purity 

of Methamphetamine Found in 

Toxic Concentration in a Civil 

Aviation Accident Pilot Fatality
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.
i

Technical Report Documentation Page 

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

DOT/FAA/AM-03/22 

4. Title and Subtitle 5. Report Date 

Distribution and Optical Purity of Methamphetamine Found in Toxic December 2003 

Concentration in a Civil Aviation Accident Pilot Fatality 6. Performing Organization Code 

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

Chaturvedi AK, Cardona PS, Soper JW, 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 tasks AM-B-00-TOX-202, AM-B-01-TOX-202, and AM-B-02-TOX-202. 

16. Abstract 

The Federal Aviation Administration's Civil Aerospace Medical Institute conducts toxicological evaluation of postmortem 

biological samples collected from pilots involved in fatal civil aircraft accidents. The submitted samples are primarily 

analyzed for the presence of primary combustion gases, alcohol/volatiles, and drugs. Related to such an evaluation, findings 

of a unique aircraft accident are described in this report. Upon colliding with terrain in weather conditions of poor visibility, 

a 1-occupant airplane was substantially damaged, with no evidence of fire. Remains of the pilot were found outside the 

crashed aircraft. Pathological examination revealed multiple blunt force injuries and vascular congestion, including subdural 

hemorrhage of the cerebral cortex. Autopsied samples—blood, brain, gastric contents, heart, liver, muscle, spleen, urine, and 

vitreous fluid—were submitted for toxicological analysis. The fluorescence polarization immunoassay disclosed 8.0 μg/mL 

amphetamines in urine. Subsequent gas chromatographic/mass spectrometric confirmatory analysis determined the presence 

of methamphetamine (1.134 μg/mL in blood and 59.171 μg/mL in urine) and amphetamine (0.022 μg/mL in blood and 

1.495 μg/mL in urine). Both amines were present in all the submitted sample types, except for amphetamine, which was 

detected neither in vitreous fluid nor in muscle. The amount of methamphetamine found in gastric contents was 575-fold 

higher than that of amphetamine. Stereochemical analyses of gastric contents, blood, and urine using a chiral probe, (S)-(–)-

N-(trifluoroacetyl)prolyl chloride, indicated that methamphetamine detected in the sample types was not optically pure. In 

gastric contents and urine, this secondary amine's optical isomers were present in equal proportions. The enantiomeric 

excess of (+)-methamphetamine over its (–)-form was about 32% in blood. Both optical forms of amphetamine were present 

in the ratio of 1.2–1.5:1.0 in the 3 sample types. The blood methamphetamine concentration found was in the range 

sufficient to produce toxic effects. The observed variation in the ratios of amine isomer concentrations in the sample types 

would have been attributed to stereoselective metabolic and other pharmacokinetic processes. Findings of this study 

supported the conclusion of the National Transportation Safety Board that, in addition to the visibility-associated adverse 

meteorological conditions, the use of the controlled substance played a contributory role in the causation of the aircraft 

accident. 

17. Key Words 18. Distribution Statement 

Forensic Sciences, Toxicology, Methamphetamine, 

Amphetamine, Distribution, Stereochemical Analyses, Aviation 

Accident Investigation 

Document is available to the public through 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 10 

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

1

DISTRIBUTION AND OPTICAL PURITY OF METHAMPHETAMINE FOUND IN TOXIC

CONCENTRATION IN A CIVIL AVIATION ACCIDENT PILOT FATALITY

INTRODUCTION

Methamphetamine, a sympathomimetic amine (1), 

exists in optical isomeric forms. Its dextrorotatory [dextro-; 

(+)-] form has central nervous system (CNS) stimulant 

effects (1,2) and has been used in the treatment of obesity, while its levorotatory [levo-; (−)-] form has been used 

as a nasal decongestant in nonprescription inhalers (2) 

and as a precursor in selegiline manufacturing (3). The 

latter isomeric form has weaker CNS stimulant and 

greater peripheral sympathomimetic effects than those 

of the former isomeric form (1,2). Methamphetamine 

has been an abused drug of concern (4-6); and unless 

specifically excepted, its isomers are considered controlled 

substances under the “food and drugs” regulations of the 

United States (7). This secondary amine is sold in illegal drug markets in the forms of (+)-methamphetamine, 

(−)-methamphetamine, and (±)-methamphetamine (2, 

8-10). (±)-Methamphetamine, also known as racemic 

methamphetamine or methamphetamine racemate, 

consists of dextro- and levo-forms in equal proportions. 

Methamphetamine mixtures of enantiomeric excess of 

one isomeric form over another isomeric form in variable proportions are also sold in the drug markets. These 

mixtures could be easily prepared by physically mixing 

dextro- and levo-forms together or by adding either of the 

isomers to methamphetamine racemate or vice versa.

In biological samples, a legitimate source of (−)-

methamphetamine could be nasal inhalers or selegiline 

(11-13) and of (+)-methamphetamine could be legally 

prescribed Desoxyn® (14). Methamphetamine is primarily biotransformed by N-demethylation to the active 

metabolite amphetamine (2,15). Biotransformation of 

optical isomers of methamphetamine and amphetamine is stereospecific—dextro-isomers of these amines 

are metabolized at faster rates than their corresponding 

levo-isomers (2,15-17).

Several methamphetamine-abuse-associated cases of 

violent and irrational behaviors (blood concentration: 

0.15–0.56 μg/mL), of erratic driving (blood concentration: 0.05–2.6 μg/mL), and of deaths with traumatic 

injuries (blood concentration: 1.4–13 μg/mL) have been 

reported in the literature (2,18,19). Blood concentrations 

of methamphetamine in the range of 0.09–18 μg/mL 

have been documented in fatal poisonings attributed to 

overdoses of this amine (2,20). Toxic and lethal levels of 

methamphetamine in blood have been considered to be 

0.6–5.0 μg/mL and 0.23 μg/mL, respectively (21,22). Although toxic and lethal blood levels of methamphetamine 

have been well documented, its stereospecificity was not 

established in those cases—it was not known whether 

methamphetamine found in those cases was dextrorotatory, levorotatory, or optical isomeric mixtures of equal 

or non-equal proportions. Knowing the stereospecific 

composition of methamphetamine in biological samples 

could assist in concluding the source of the amine and 

in accurately predicting the level of the adverse effects 

caused by the substance.

In the present study, toxicological findings of a unique 

civil aviation accident pilot fatality involving methamphetamine are described. Additionally, distribution 

and stereospecificity of this amine and its metabolite 

—amphetamine—were determined in the various submitted postmortem biological samples. Also, included are 

pathological findings and aircraft accident case history.

Case History

A Cessna airplane was substantially damaged upon 

colliding with terrain at approximately 1700 hours in an 

isolated area. The only occupant, the pilot, was fatally 

injured. In the general area of the accident, heavy snow 

conditions were observed, and the visibility was 0.5 mile 

(805 m) with fog and scattered overcast. An airman’s 

meteorological information for mountain obscuration 

was in effect in the area at the time of the accident. There 

was no sign of fire in the crash. A strong odor of gasoline 

was present near the wreckage. Seat belts and shoulder 

harnesses were not buckled. The remains of the pilot were 

found on the ground outside the airplane. The autopsy 

was conducted 69 hours after the accident.

Pathology

The pilot was a well-developed and well-nourished 44-

year-old Caucasian male, weighing approximately 180 lbs 

(82 kg). The length of the body was about 70 in (1.8 m). 

The body was cold. Anatomical examination of the body 

revealed multiple blunt force injuries—bone fractures and 

internal lacerations—in the chest, abdomen, and lower 

back; and a small focus of subdural hemorrhage over the 

left parietal region of the cerebral cortex. Lacerations, 
2 

contusions, and abrasions were noted, practically all over 

the body. Vascular congestion was found in the kidneys, 

lungs, heart, spleen, and left frontal/parietal cortex. There 

was no evidence of medical therapy. Autopsied biological samples—blood, brain, gastric contents, heart, liver, 

muscle, spleen, urine, and vitreous fluid—were submitted 

to the Federal Aviation Administration’s Civil Aerospace 

Medical Institute in Oklahoma City, OK, for toxicological evaluation.

MATERIALS AND METHODS

Materials

All reagents and solvents were of analytical grade and 

of the highest available purity. Reagents, solvents, standards, internal standards, and derivatizing agents were 

obtained from commercial sources. Bovine whole blood 

used for the preparation of calibrators and controls was 

obtained from a local slaughterhouse. The AxSyM®

Amphetamine/Methamphetamine II assay kit and controls for the screening of amphetamine/methamphetamine 

in urine were purchased from Abbott Laboratories (Abbott Park, IL). This screening assay utilizes fluorescence 

polarization immunoassay (FPIA) technology. During 

the assay, 0.5, 1.5, and 4.0 μg/mL controls of (+)-amphetamine were used. Cross-reactivity of this assay with 

(−)-amphetamine, (+)- and (−)-methamphetamines, and 

other structurally related molecules has been reported 

by the kit manufacturer. Standards of (+)- and (−)-amphetamines and of (+)- and (−)-methamphetamines were 

obtained in methanolic solutions from Alltech-Applied 

Science Labs (State College, PA). Internal standards were 

supplied as racemic mixtures, (±)-amphetamine-d8 and 

(±)-methamphetamine-d8, by Cerilliant Corporation 

(Austin, TX). Pentafluoropropionic anhydride (PFPA) 

was purchased from Pierce Chemical Company (Rockford, IL). The chiral probe used in resolving enantiomers 

of amphetamine and methamphetamine was (S)-(−)-N-

(trifluoroacetyl)prolyl chloride (TPC). Obtained from 

Aldrich Chemical Company, Inc. (Milwaukee, WI), this 

probe was 97% enantiomerically pure and was supplied 

as 0.1 M solution in dichloromethane. TPC has previously been used as a probe in the stereospecific analysis 

of amphetamines (11-13).

Analytical Toxicology

The submitted pilot fatality biological samples were 

analyzed for the presence of combustion gases (carbon 

monoxide and hydrogen cyanide), alcohol/volatiles, 

and drugs—which include a wide range of prescription, 

nonprescription, and abused drugs (23-27). These exogenous substances in the biological samples were analyzed 

according to established standard laboratory procedures, 

including screening and confirmatory/quantitative 

analyses (24). Carbon monoxide in blood was analyzed 

spectrophotometrically as carboxyhemoglobin (28) 

and hydrogen cyanide colorimetrically as cyanide (29). 

Alcohol/volatile analysis in vitreous fluid was performed 

by headspace gas chromatography. The urine sample was 

screened for abused drugs by FPIA and for prescription 

and nonprescription drugs by high-performance liquid 

chromatography and by gas chromatography/mass spectrometry (24).

Gas Chromatograph/Mass Spectrometer

An Agilent gas chromatograph (Model 6890)/mass 

spectrometer (Model 5973) system (Agilent Technologies, 

Wilmington, DE) was used during the confirmatory/ 

quantitative and stereochemical analyses of methamphetamine and amphetamine. A crosslinked 100% methylsiloxane wall-coated open tubular column (12.5 m ×

0.2-mm i.d.; 0.33-μm film thickness) was used in the 

gas chromatograph/mass spectrometer (GC/MS) system. 

Helium was the carrier gas with a constant flow of 1 mL/

min. The injection volume was 1 μL in the splitless mode, 

with a splitless time of 0.5 min. The injector temperature 

was maintained at 250°C, and the transfer line was set at 

280°C. The acquisition was in the electron impact mode 

of ionization using selective ion monitoring.

Confirmatory/Quantitative Analyses

Extraction

Based on the urine FPIA screening finding, each of the 

submitted biological sample types was analyzed for the 

confirmation and quantitation of amphetamines. This 

analysis was achieved by the solid phase extraction (SPE) 

technique using Bond Elut CertifyTM columns (Varian 

Sample Preparation Products, Harbor City, CA).

Depending upon the possible anticipated amounts 

of the amines present in a particular sample type and 

its availability, different amounts of samples were used 

for extraction. This approach was taken to ensure that 

amine concentrations in the respective sample types fall 

within the range of the calibration curves. Amounts of 

samples used were 0.30, 1.0, and 3.0 mL blood; 0.030 

and 3.0 mL urine; and 0.15 and 0.30 mL vitreous fluid. 

Homogenates of solid tissues were prepared in 1% NaF 

aqueous solution. Amounts of homogenates used for 

analysis represented 0.030, 0.30, and/or 1.0 g of a particular tissue type. Two gastric content aliquots in the 

amounts of 0.030 and 0.30 g were separately diluted with 

suitable amounts of deionized water, homogenized, and 

centrifuged at 770 × g for 5 min to obtain their respective 

homogeneous supernatants.
3

Aliquots (3.0 mL or less) of each calibrator, control, 

fluid specimen, diluted specimen, homogenate, and gastric 

content supernatant in separate 16 mm × 100 mm screwcap culture glass tubes were separately spiked with 1.0 mL 

of 400 ng/mL aqueous solutions of internal standards, 

(±)-amphetamine-d8 and (±)-methamphetamine-d8. With 

those samples wherein less than 3.0 mL of aliquots were 

used, their volumes were brought to 3.0 mL by adding 

deionized water. To each of the tubes containing blood 

and homogenate, 10.0 mL of cold acetonitrile (4°C) 

was added to precipitate protein: Acetonitrile was not 

added to the tubes containing urine, vitreous fluid, and 

gastric content supernatant samples. All acetonitrile 

containing tubes were capped, shaken for 1 min, and 

placed on a rotary extractor until the mixture reached 

ambient temperature and then centrifuged at 770 × g

for 5 min. Acetonitrile supernatants were decanted to 16 

mm × 125 mm screw-cap culture glass tubes and were 

evaporated to less than 1 mL at 40°C, in a water bath, 

by using a stream of nitrogen. Subsequently, 2.0 mL of 

0.10 M phosphate buffer (pH 6.00) was added to each 

tube, including those tubes that contained urine, vitreous 

fluid, and gastric content supernatant samples.

Under a positive pressure manifold, the SPE columns 

for each of the tubes were first conditioned with 2.0 mL 

of methanol, followed by 2.0 mL of the phosphate buffer. 

The previously buffered sample solutions were transferred 

onto the conditioned columns and allowed to pass through 

the columns. Then, the columns were rinsed with 1.0 

mL of 1.0 M acetic acid and dried with 25 lb of pressure 

for 5 min. Subsequently, 6.0 mL of methanol was passed 

through the columns, and they were then allowed to dry 

for 2 min with 25 lb pressure. Analytes were eluted from 

the columns into 10-mL screw-cap conical glass tubes by 

using 4.0 mL of 2% NH4OH in ethyl acetate. Hydrogen 

chloride was bubbled through the collected eluates to 

minimize the loss of amphetamines.

PFPA Derivatization

The obtained eluates were evaporated to dryness at 40°C, 

in a water bath, by using a stream of nitrogen. To each tube 

was added 50.0 μL of ethyl acetate, and each tube was 

vortexed for 15 sec. Subsequently, 50.0 μL of PFPA was 

added to the tubes. These reaction mixtures were incubated 

for 20 min at 70°C in a heating block, allowed to cool to 

ambient temperature, and evaporated to dryness in a 40°C 

water bath by purging with nitrogen, followed by the addition of 50.0 μL of ethyl acetate for reconstitution. The 

reconstituted mixture from each tube was then transferred 

into 200-μL inserts of autosampler vials, and 1.0 μL of the 

mixture from the inserts was injected onto the GC/MS 

system for characterization, confirmation, and quantitation 

of the derivatized products of amphetamines.

GC/MS Conditions for PFPA Derivatives

The initial GC oven temperature of 70oC was increased to 150°C at 15°C/min and then to 290°C at 

40°C/min with no hold times. The retention times (±

0.01 min) for PFPA derivatives of (±)-amphetamine-d8, 

(±)-amphetamine, (±)-methamphetamine-d8, and (±)-

methamphetamine were 3.72, 3.75, 4.54, and 4.57 min, 

respectively. The ions (m/z) monitored were 193, 126, 

and 96 for PFPA-(±)-amphetamine-d8; 190, 118, and 

91 for (±)-amphetamine; 211, 163, and 123 for PFPA-

(±)-methamphetamine-d8; and 204, 160, and 118 for 

PFPA-(±)-methamphetamine. The first of the 3 ions of 

each of these analytes was used as their quantifying ions, 

while the remaining 2 ions of each of the analytes were 

used as their qualifying ions. Calibration curves for amphetamine and methamphetamine in bovine blood were 

constructed. Calibrators of both amines for obtaining 

the curves were 0.025, 0.050, 0.100, 0.200, and 0.400 

μg/mL. The regression analysis disclosed that both curves 

were linear (r ≥ 0.9990) in the stated concentration range. 

Since PFPA is a non-chiral compound, the stereospecificity of amphetamines in the submitted samples could not 

be established by this analysis.

Stereochemical Analyses

Extraction

To establish enantiomeric purity, amphetamines were 

extracted from re-accessioned gastric content, blood, and 

urine samples by the liquid-liquid extraction method and 

were then derivatized by the TPC chiral probe. Because 

amphetamines were present in varying amounts in these 

sample types, different amounts of samples were used for 

the extraction so that amine concentrations fall close to 

a 0.200-μg/mL control, the mid-point concentration on 

the quantitative analysis calibration curve for each amine. 

A mixture control, in bovine blood, consisting of 0.200 

μg (200.0 μL of 1.0 μg/mL) of (+)-amphetamine, (−)-

amphetamine, (+)-methamphetamine, and (−)-methamphetamine each was analyzed. Individual controls of each 

of these analytes at 0.200 μg (200.0 μL of 1.0 μg/mL) in 

bovine blood were also separately analyzed. Bovine blood 

was used as a negative control. As was done during the 

quantitative analyses, 2 supernatants of the gastric content 

sample in deionized water were prepared; 200.0 μL of 

the supernatants represented 0.010 and 0.820 g of gastric 

contents. The 0.010-g gastric content supernatant was 

used for methamphetamine analysis, while the 0.820-g 

gastric content supernatant for amphetamine analysis. 

Amounts of blood and urine used were also 200.0 μL. 

Two-hundred-μL aliquots of each control and sample 

were transferred into separate 16 mm × 150 mm screwcap glass test tubes. To each tube were added 1.0 mL of 

400 ng/mL aqueous solution of the internal standards 
4 

(±)-amphetamine-d8 and (±)-methamphetamine-d8, 4 

drops of concentrated NH4OH, and 10.0 mL of n-butyl 

chloride. Each tube was vigorously shaken for 2 min 

and then centrifuged at 770 × g for 5 min. The organic 

phase from the tubes was transferred into 16 mm × 150 

mm screw-cap glass test tubes. After adding 4.0 mL of 

1.0 N HCl to each of the tubes, they were vigorously 

shaken for 2 min and centrifuged at 770 × g for 5 min. 

The separated organic phase was discarded, and 1.0 mL 

of concentrated NH4OH was slowly added to each of 

the tubes, kept in ice, while swirling. After 15 min, tubes 

were vortexed for 15 sec, and 4.0 mL of chloroform was 

added to each tube. Subsequently, tubes were shaken and 

centrifuged, and the upper aqueous phase from each tube 

was removed by aspiration. The chloroform phase from 

each tube was decanted into 10-mL screw-cap conical 

glass tubes, and hydrogen chloride was bubbled through 

each chloroform solution.

TPC Derivatization

The hydrogen chloride-bubbled chloroform solutions 

were evaporated to dryness at 40°C, in a water bath, by 

using a stream of nitrogen. Tubes were allowed to come 

to room temperature, and then 50.0 μL of the TPC solution was added to each tube. After a brief mixing, the 

obtained reaction mixtures were incubated for 30 min 

at 70°C in a heating block, allowed to cool to ambient 

temperature, and evaporated to dryness in a 40°C water 

bath by purging with nitrogen. To each tube was added 

50.0 μL of ethyl acetate to reconstitute the residue. All 

tubes were vortexed for 15 sec, and then the ethyl acetate 

solutions were transferred into 200-μL inserts of autosampler vials and 1.0 μL of each solution was injected 

onto the GC/MS system.

GC/MS Conditions for TPC Derivatives

The GC oven temperature was set at 70°C; it was then 

increased to 290°C at a rate of 30°C/min with a final hold 

time of 2.67 min. The retention times (± 0.01 min) of the 

TPC-derivatized diastereomers, (−)-amphetamine-d8, (−)-

amphetamine, (+)-amphetamine-d8, (+)-amphetamine, 

(−)-methamphetamine-d8, (−)-methamphetamine, (+)-

methamphetamine-d8, and (+)-methamphetamine, were 

4.94, 4.95, 5.01, 5.02, 5.40, 5.42, 5.45, and 5.47 min, 

respectively. The ions (m/z) monitored were 240, 241, and 

242 for TCP-(−)-amphetamine-d8 or TCP-(+)-amphetamine-d8

; 237, 238, and 239 for TCP-(−)-amphetamine or 

TCP-(+)-amphetamine; 258, 259, and 260 for TCP-(−)-

methamphetamine-d8 or TCP-(+)-methamphetamine-d8; 

and 251, 252, and 253 for TCP-(−)-methamphetamine 

or TCP-(+)-methamphetamine. The first of the 3 ions 

of each of these analytes was used as a quantifying ion 

for the determination of enantiomeric percentages, while 

the remaining 2 ions of each of the analytes were used 

as qualifying ions. 

The enantiomeric percentages were determined by 

separately calculating ratios of TCP-(−)- and TCP-(+)-

amine peak areas to the peak areas of respective isomeric-d8

internal standards and then dividing each of the obtained 

ratios with the sum of both ratios as expressed in the 

following formula.

[{PATCP-(− or + )-amine ÷ PATCP-(− or + )-amine-d8} × 100] ÷

[{PATCP-(−)-amine ÷ PATCP-(−)-amine-d8} + {PATCP-(+)-amine ÷

 PATCP-(+)-amine-d8}]

Where, PA = peak area of amphetamine or methamphetamine TCP derivatives. 

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5

RESULTS AND DISCUSSION

Analysis of the submitted blood sample failed to disclose 

the presence of carbon monoxide, cyanide, or ethanol in a 

detectable amount. However, the FPIA technology-based 

assay revealed the presence of 8.0 μg/mL amphetamines 

in urine. Because of cross-reactivity, this screening assay 

does not correctly distinguish optical isomeric forms of 

amphetamine or methamphetamine and also does not 

distinguish amphetamine from methamphetamine. Since 

the former is a metabolite of the latter and the cross-reactivity with other abused drugs structurally similar to 

amphetamine has been reported with the FPIA assay, a 

more specific alternate GC/MS-based analytical method 

was used for confirming the FPIA analytical result. The 

GC/MS analysis revealed the presence of methamphetamine in all the submitted sample types and of amphetamine in all the sample types but in vitreous fluid and 

muscle (Table I). This information suggested that methamphetamine and its metabolite—amphetamine—were 

distributed throughout the body, including the brain, 

of the pilot. As expected, the higher amounts of amines 

were present in urine, liver, or spleen than the amounts 

in vitreous fluid or in muscle—in fact, amphetamine 

was not detected in these 2 sample types. The levels of 

amphetamine were significantly lower in all the sample 

types than the levels of methamphetamine in the corresponding sample types. The blood methamphetamine 

level of 1.134 μg/mL was in the range sufficient to produce toxic effects, including erratic behavior (2,18,19). 

This level overlapped the lethal concentration range of 

methamphetamine (0.09–18 μg/mL) reported in the literature (2,20). Although 0.022 μg/mL of amphetamine 

in blood is a subtherapeutic concentration (15,21), its 

contributory role cannot be ruled out in enhancing the 

overall effects of methamphetamine in the deceased. The 

575-fold higher amount of methamphetamine found in 

the gastric contents than that of amphetamine suggested 

that methamphetamine might have been taken orally. 

This conclusion is further supported by the fact that the 

methamphetamine to amphetamine concentration ratio 

(575) in the gastric contents is drastically higher than the 

ratios (39.6–62.4) in the 6 different body compartments 

wherein amphetamine was metabolically generated (Table 

I). Basic drugs have a tendency to accumulate in the 

stomach because of the pH gradient between plasma and 

gastric juice (30). Therefore, the relatively small amount 

of amphetamine found in the gastric contents was potentially the result of the low-pH associated ion trapping of 

the methamphetamine metabolite in the stomach. Since 

only a portion of the gastric contents was submitted for 

the analysis, the total amounts of both amines present 

in the stomach could not be calculated.

Unless specifically excepted, optical isomers of amphetamine and of methamphetamine are considered controlled 

substances under the U.S. regulations of food and drugs 

(7). However, (−)-methamphetamine and its metabolite 

could legitimately be found in biological samples of individuals who use nasal inhalers or the anti-Parkinson’s 

medication selegiline (11-13). Therefore, there is a genuine 

need for analytically determining stereospecificity—levorotatory, dextrorotatory, or optical isomeric mixture—of 

methamphetamine and its metabolite (amphetamine) 

for establishing the degree and type of biological effects 

and the source of the amines. Such determination is of 

significance because a particular optical isomer may produce a selective biological effect, may be more potent 

than the other isomer (1,2,15), or could be considered 

a controlled substance (7).

Blood methamphetamine concentrations reported in 

the literature do not specifically mention stereospecificity of the amine (2,18-20). The amine could be in the 

(+)- or (−)-optical form or could be a mixture consisting 

of both optical forms in equal or non-equal proportions. 

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6 

Since pharmacological effects of methamphetamine are 

stereospecific [(+)-form has stronger central stimulant 

effects than the (−)-form], a racemic methamphetamine 

[(±)-methamphetamine]—that is, (+)-methamphetamine 

and (−)-methamphetamine in equal proportions—would 

have pharmacological effects falling somewhere between 

the degree of effects caused by each of the optical isomers 

alone. Therefore, the concentration of the amine found in 

the biological samples could not necessarily be accurately 

correlated with the degree of toxicity, provided the stereospecificity of methamphetamine has been established.

Stereochemical analyses of the gastric contents, 

blood, and urine disclosed that methamphetamine and 

amphetamine were present in the samples in an optical 

mixture form (Table II). As is given in the table, (+)- and 

(−)-forms of methamphetamine were present in almost 

equal proportions in gastric contents and in urine, but the 

percent concentration ratio of (+)-methamphetamine to 

its (−)-form was 2:1 in blood. On the other hand, (+)- and 

(−)-amphetamines were present in the gastric contents, 

blood, and urine in the ratios of 1.5:1, 1.2:1, and 1.4:1, 

respectively. The biotransformation of methamphetamine 

and amphetamine is stereoselective; their (+)-form is metabolized faster than their (−)-form (2,15-17). Therefore, 

the (+)-form should be present in the lesser amounts than 

the (−)-form in various body compartments. However, 

in vivo stereoselective pharmacokinetic processes other 

than metabolism may prevent these isomers from reaching 

such an anticipated concentration pattern in a particular 

body compartment. Since biotransformation is essentially 

negligible in the stomach, the stereoselective effect of the 

process will also be minimal on the concentrations of 

optical isomeric forms of methamphetamine. Thus, the 

concentration ratio found in the gastric contents more 

likely reflect the optical purity of methamphetamine 

ingested by the deceased. However, the isomeric concentration ratios of both amines in other sample types 

could also be the resultant of the effectiveness of other 

stereoselective pharmacokinetic processes, leading to 

the different isomeric concentration ratios in different 

sample types.

Considering the isomeric ratio of methamphetamine in 

blood, concentrations of its (+)- and (−)-forms could be 

calculated as to be 0.748 and 0.386 μg/mL, respectively. 

Therefore, the net biological effect on the deceased was 

caused by both isomers, and the intensity of the effect 

would have been less than that would have been caused 

by only dextro-methamphetamine at 1.134 μg/mL. The 

possibility for reaching the levo-isomer’s concentration at 

the calculated value from a legitimate use of a non-prescription inhaler or from selegiline is remote, particularly 

in the presence of the dextro-isomer and in the absence 

of selegiline itself.

Toxicological findings suggested that the deceased took 

the racemic methamphetamine orally. This suggestion was 

supported by the presence of a relatively large amount of 

(±)-methamphetamine in the gastric contents in relation 

to amphetamine. The origin of (±)-methamphetamine 

could have been a chemically synthesized product from 

phenylacetone and N-methylformamide (2, 8-10) or a 

physically prepared product from mixing (+)- and (−)-

forms of methamphetamine in equal amounts, as these 

chemical and physical routes would produce methamphetamine racemate. The oral ingestion allowed the 

blood methamphetamine concentration to reach a level 

sufficient to produce some degree of central stimulant 

effects, causing performance impairment. Findings of 

this study supported the National Transportation Safety 

Board’s determination of factors of the accident as “meteorological conditions obscuring the pilot’s visibility and 

incapacitation due to illegal substances.”

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1

 This document can be obtained through the National Technical 

Information Service, Springfield, Virginia 22161. It is also available 

at the CAMI Web site: www.cami.jccbi.gov.