Comparison and classification of methamphetamine
seized in Japan and Thailand using
gas chromatography with liquid-liquid extraction
and solid-phase microextraction
Kenji Kuwayama et al.
2
INTRODUCTION
3
Methamphetamine
Caffeine
Amphetamine
4
Methamphetamine
Amphetamine
5
eat
 inject
 smoke
 inhale

6
H
N
Methamphetamine
OH
O
1-phenyl-2-propanone (P2P)
H
N
Ephedrine compound
7
 Presumptive Test
 Thin Layer Chromatography (TLC)
 Gas Chromatography Flame Ionization Detector (GC-FID)
 Gas Chromatography Mass Spectrometry (GC-MS)
 High Performance Liquid Chromatography (HPLC)
 Fourier Transform Infrared (FTIR) Spectroscopy
ทีม่ า : United Nations Office on Drugs and Crime
8
Two GC method for MA impurity profiling
NRIPS
ONCB
The national Research Intitute
of Police Science
The Office of the Narcotics
Control Board
JAPAN
THAILAND
9
JAPAN
THAI
LLE
10
Chromatograms obtained using the ONCB method
tablet
THAI
crystal
11
Chromatograms obtained from an MA crystal
ONCB
NRIPS
12
The aim of this study
• Improve the analytical method for profiling MA
impurity
• Compare and classification MA crystals seized in
different countries
• Information in criminal investigation ; traffic routes ,
the source of supply and relationships between seizures
13
Method
Liquid-Liquid Extraction
( LLE )
Solid-Phase Microextraction
(SPME )
GC-FID & GC-MS
14
Technique LLE
solution
sample
GC
Shaking
centrifuge
Organic layer
15
15
Technique SPME
SPME fiber
Head space
SPME fiber
GC
16
Technique GC-FID
FID
17
Technique GC-MS
18
MATERIALS
AND
METHODS
19
Reagents and chemical
1. MA.HCl crystals
seized in Japan (69) and Thailand (42)
2. Std. d-MA.HCl
3. l-ephedrine.HCl
4. dl-Dimethylamphetamine.HCl
5. cis-1,2-dimethyl-3-phenylaziridine
6. n-Alkanes : Internal Standard
7. Solvents
8. SPME holder and fiber coated with DVB/CAR/PDMS
9. Inlet liner for SPME
20
Gas chromatographic analysis
1.GC-FID
Agilent – 6890
Auto inject : 7683
21
2.GC-MS
Agilent – 6890
Agilent 5973N MSD
22
COLUMN
GC-FID & GC-MS
DB-5 capillary column
( 30 m. x 0.32 mm. x film
thickness 1.0 µm. )
23
LLE procedure ( NRIPS )
buffer solution1 mL
Shaking
MA.HCl 50 mg
GC
Istd.
0.5 mL Ethyl acetate + Istd.
Centrifuge 3000 rpm
5 min
n-decane (C10,IS1) n-pentadecane (C15,IS2) n-eicosane (C20,IS3)
24
24
n-octacosane (C28,IS4)
0.02 mg/ml
SPME procedure
Headspace
at 85OC
35 min
SPME fiber
MA.HCl 10 mg
SPME fiber
GC
25
Condition: GC-FID & GC-MS
Initial temperature : 50 0C held 1 min. increase of
10 0C/min. to 300 0C held 10 min.
0
240 C
Inject temperature :
Detector temperature : 300 0C
Carrier gas
: He (g) flow rate 2 mL / min.
Injection
: 1 µL splitless mode
Condition for SPME were the same as those for LLE
26
RESULTS
27
Data processing for LLE & SPME
• Peak data integrated by Chemstation software
• Processed using the Drug Micro-Component
Analysis & Comparison System (DMCPS)
Calculation of Euclidean distances
Cluster analysis by Ward method
28
Data processing for LLE
Istd.
29
Typical chromatograms obtained from MA crystal using NRIPS
Istd.
Ephedrine (pseudoephedrine)
30
Data processing for SPME
31
Typical chromatograms obtained samples by SPME
Empty vial without MA
32
Typical chromatograms obtained samples by SPME
33
1.Optimization of analytical procedure
LLE
ONCB
optimized for the analyzed of MA tablet
NRIPS
optimized for the analyzed of MA crystal
considered more efficient than the ONCB
34
2.Cluster analysis of sample seized in Japan and Thailand
MA crystals seized
Japan = 69 sample
Thai = 42 sample
NRIPS method
Cluster analysis
Fourteen characteristic impurity peak
35
2.Cluster analysis of sample seized in Japan and Thailand
NRIPS method
No.sam. (J/T) 15/19
25/0
17/15
36
12/8
Typical chromatograms of MA samples in each group using the NRIPS
High purity
Group A
Group B
1,3-dimethyl-2-phenyl naphthalene
1-benzyl-3-methylnaphthalene
37
Typical chromatograms of MA samples in each group using the NRIPS
cis-1,2-dimethyl-3-phenylaziridine
Group C
Ephedrine (pseudoephedrine)
Group D
38
3.Comparison and classification of sample in the high purity group by SPME
When MA crystals were extracted with SPME and analyzed by
GC (SPME ), the chromatograms had many impurities while the
chromatograms in case of LLE had few impurities
MA
Many impurity
Characteristic peaks for data processing
seven peaks
39
3.Comparison and classification of sample in the high purity group by SPME
15/19
SPME
NRIPS
40
3.Comparison and classification of sample in the high purity group by SPME
Chromatograms were distinguished clearly from
SPME method , whereas in the case of LLE it was
difficult to compare and classify samples in the highpurity group because there were so few impurities.
The SPME method enables us to compare and
classify high-purity MA
41
CONCLUSION
42
Compared
LLE
ONCB
NRIPS
Superior for detecting and
separating MA impurities
14 peaks
characteristic
Classified into
four groups
43
LLE
SPME
Effective for comparing high-purity MA
because it detected many characteristic peaks
44
45
QUESTION
46