Forensic Science International, 50 (1991) 47-55

Elsevier Scientific Publishers Ireland Ltd.

COMPARATIVE

ANALYSIS

OF ILLICIT

47

HEROIN

SAMPLES

M. CHIAROTTIa, N. FUCCIa and C. FURNARI

aZstitutodi Medic&a Legale-Universita Cattolica de1 Sacro Cwe di Roma, L. go F. Vito 1 and
bZstitutodi Medic&a Legale ZZUniversita di Rma Via 0. Raimondo, Roma (Italy)
(Received April 2nd, 1990)
(Final revision received December 5th, 1990)
(Accepted February 26th, 1991)

Summary
The comparative analysis of street heroin samples is still an object of scientific discussion. A combination of a wide range of analytical techniques is necessary to obtain a valid amount of information
about the sample composition, and it is impossible to define an unique analytical approach. We suggest a complete analytical sequence based on analysis of volatile compounds, opiates, diluents,
adulterants and metals, by head space gas chromatography (HS/GC), gas chromatography mass
spectrometry (GCIMS), thin layer chromatography
(TLC), high pressure liquid chromatography
(HPLC) and atomic absorption (AA) using a sample amount as low as 50-100 mg. The outlined procedure can be successfully applied to routine work, thus obtaining suitable information about a samples chemical composition. This helps to attribute or exclude common sources of separate specimens.
Results obtained on 33 street heroin samples confiscated in the metropolitan area of Rome are listed.
Key woro!s:Heroin; Comparative

analysis; Common origin

Introduction

The comparative analysis of two or more heroin samples is frequently required

by the forensic toxicologist in order to establish their probable common source.
In this field, many papers have been reported by various authors with the aim
of suggesting suitable analytical procedures to reach this goal [l-3]. Consequently, a variety of methods may be used for the comparative analysis of street
heroin samples and each of these procedures uses different parameters such as
the specific qualitative and quantitative determination of related narcotics (codeine, acetylcodeine, monoacetylmorphine,
noscapine, papaverine, etc.) [4-61.
Besides diluents, adulterants and solvents are also identified, mainly with the
aim of obtaining more information about the chemical composition in order to
identify the source of supply or origin of illicit samples [7-g]. A wide range of
information is needed for comparative purposes and the relative amount of some
components may change in heroin samples during the illegal drug traffic, or they
could be modified under a particular analytical process [lo-121. Other additional
investigations may be necessary in order to provide a valid sample comparison.
A comprehensive sequential analytical procedure is not still defined. We suggest
0379-0738/91/$03.50
0 1991 Elsevier Scientific Publishers
Printed and Published in Ireland

Ireland Ltd.

48

an operative procedure involving, by means of combinations of different techniques, the comprehensive identification of heroin components. This may be useful
information that leads to relating an illicit heroin sample to another batch of
heroin of known origin.
Materials and Methods

All solvents were of analytical grade. Pure chemical standards of heroin,

papaverine, codeine, acetylcodeine, ethaverine, phenobarbital, methaqualone,
caffeine, narcotine, were purchased from Supelco (Bellafonte, Palo Alto, CA).
Standards of acetylthebaol, 3-6 dimethoxyphenanthrene-4,5
epoxide and
thebaol, were kindly donated by Dr. H. Huizer (Gerechtelijk Laboratorium Van
Het Ministerie Van Justicie, Rijswijk, The Netherlands). 3-Monoacetylmorphine
and 6-monoacetylmorphine were synthesized according to Welsh 1131.
Thirty-three heroin samples seized in the metropolitan area of Rome were
analyzed by head space gas chromatography
(HS/GC), thin layer
chromatography (TLC), high pressure liquid chromatography (HPLC), gas
chromatography mass spectrometry (GUMS) and atomic absorption (AA) according to the sequence schematized in Table 1 and operating as indicated therein.
An amount of a sample (50-100 mg) was transferred to a lo-ml vial for
preliminary analysis of volatile solvent residues (HS/GC). The vial was sealed
with a flange type rubber stopper and aluminium seal and then placed into a head

TABLE 1
ANALYTICAL SEQUENCE PROPOSED FOR ANALYSIS OF ILLICIT HEROIN SAMPLES
(ANALYTICAL CONDITIONS ARE REPORTED IN TEXT)
50-100
determination

mg samples
of volatile compounds

1
HS/GC

I
+ 5 ml of methanolic solution containing an internal standard (for quali-quantitative
of opiate and adulterants)

I
TLC - GC - GUMS

1
determination

of diluents

1
HPLC

I
trace metal detection

1
AA

determination

49

space autosampler for gas chromatographic analysis. The analysis was carried
on Porapak Q column (2 m x 4 mm) at 16OC, after 15 min equilibration time
at 60C. The next successive steps were the analysis of opiate and related compounds, adulterants and diluents, by gas chromatography, thin layer
chromatography and high pressure liquid chromatography. These analyses were
performed after the addition of 5 ml of methanolic solution of ethaverine (2
mg/ml) as internal standard to the vial.
TLC

TLC (for the detection of fluorescent compounds such as thebaol, acetylthebaol

and phenanthrene derivates of thebaine) was carried out according to H. Huizer
[14] using silica gel as stationary phase and n-hexane:dichloromethane:methanol
(0.75% diethylamin) (72:20:5) as mobile phase. After developing, the plate was
observed under a UV lamp at 360 nm.
GC

CC analysis (for quali-quantitative determination of opiates and adulterants)

was performed on OV 101 capillary column (15 m, 0.1 pm film thickness) using
a temperature program between 60 and 280C with 3Clmin linear increments,
equipped with an ion trap detector (electronic impact, 70 eV) operating in total
ion current between 50 and 400 a.m.u.
HPLC

HPLC (for diluent analysis) was carried out using a Supelcosil-LC-(R)-urea (5

pm) column and acetonitrile:water (75:25) as mobile phase at a flow rate of 1
ml/min, with UV detection at 190 nm.
Finally, the methanolic solution was evaporated to dryness and analyzed by
AA for trace metal detection. For this purpose, 6 N nitric acid was added to the
residue and the resulting solution was analyzed by atomic spectrometer (Mod.
Varian) equipped with a hollow cathode lamp (284.3 nm) for iron and (213.9 nm)
for zinc detection.
Results and Discussion

As previously described [9], volatile substances were always present and ageing of the sample did not seem to impair their detection. One to five solvents
were observed for each sample. Acetic acid, acetone, diethylether, benzene and
alcohols, such as methanol and ethanol, were detected. In some cases, the
presence of acetaldehyde indicated a probable use of ethylidene diacetate during
the heroin synthesis process [15].
The data resulting from the proposed analytical protocol are schematically
reported in Table 2 and they can be summarized as follows. Broad distribution
of the values concerning purity of heroin (ranging from 5 to 96%) was observed.
Adulterants were present in 50% of the samples and among these caffeine,
phenobarbital and methaqualone, were the most frequently identified. Diluent
analyses both qualitative and quantitative, easily carried out by HPLC using an

50

TABLE 2
THE DATE RESULTING FROM ANALYSIS OF 33 HEROIN SAMPLES SEIZED IN THE
METROPOLITAN AREA OF ROME
Sample

HS-GC

GC

TLC

HPLC

Heroin (%) Adulterants

1
2
3
4
5
6
7
8
9

10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33

4-5-6
l-5-6
4-6
6
l-4-6
1-6
6
l-4-5-6
l-2-4-5-6
l-3-5-6
3-4-5-6-7
l-4
l-3-4-5-6
1-6
l-5-6
l-3-6
6
l-4-5-6
l-3-6
l-3-6
6
4-5-6
l-3-4-6
2-3-4
4-6
4-6
l-5-6-7
3-4-5-6-7
4-5-6
l-3-5-6-7
l-2-4-6
l-4-6
l-4-5-6

88
67
9
5
79
73
96
65
17
15
38
52
78
85
54
85
94
29
40
15
53
48
15
86
58
70
32
29
40
26
40
79
65

Pro
PhelMet
Caf/Phe
Pro

Caf
Caf
CaflPhe
Caf/Phe/Met
Caf
Met
PhelMet
Caf/Phe/Met
Phe
Phe

AB.
A
AB
ABC
A
A
AB
AB
A
A
AB
AB
A
AB
ABC
AB
AB
AB
A
AB
AC
A
ABC

G
S
S
L
G
S
G
G
S
S
G
L
G
S
S
G
G

AA
Fe

Zn

87
72
30
20
48
30
70
38
63
93
170
38
79
52
27
30
48
70
104
18
186
115
188
34
161
189
218
149
69
50
77
48
37

57***
43
27
20
48
25
53
30
62
89
26
21
54
48
34
42
47
30
67
26
70
103
138
55
24
32
167
49
30
73
48
50
30

*l = methanol; 2 = acetaldehyde; 3 = ethanol; 4 = acetone; 5 = diethylether; 6 = acetic acid; 7

= benzene.
**Fluorescence A = Rf 0.0-0.3; B = Rf 0.31-0.6; C = Rf 0.61-0.95.
***ppm.
S = saccharose; G = glucose; L = lactose; Pro = procaine; Phe = phenobarbital; Met = metaqualone; Caf = caffeine.

51

glucose.

Fig. 2. High performance

liquid chromatographic

pattern

of sugars in an illicit heroin sample (analytical conditions are reported in text). Rt = 7.91

53

r-I!!
-%
--

__If--L

-%
-- _
I

--L
-%

-.&

32
s

alluutwan

WEN
Libr Skh
cxiw#:
88 l@4@4
;6&xp
leetl Sanple

94

4 = heroin; 5 = papaverine;

ired:Feb-W198814:04:26t
A9cI
5 a1
Intemity 728
scmlderssl

Fig. 4. Finger print of opiates and adulterants in heroin samples. 1 = caffeine; 2 = phenobarbital; 3 = monoacetylmorphine;
6 = noscapine.

M-84-198814:04:26
PeaksFad: 3

14:84:26
8o,~f~202~5rl
9 iml: Feb-Otl9RR
Scm:754 Id:3633
C l2:31 RIG:lW=368919

fin&r
DataFile; KMEN hired:
ScW 548to 587 Pe* Pmflle Fib (7)

RJ

~~:25#-754

55

sanples
Fig. 5. Quantitative
see the text).

analysis of Fe and Zn for each sample of illicit heroin (for analytical conditions

aminic resin as stationary phase, showed that sucrose and glucose are more frequently used as diluents. Under our conditions, a complete resolution of five
sugars commonly employed as diluents by drug trafficants was achieved in 15
min (Fig. 1). A chromatographic
pattern of sugars in a real sample is shown in
Fig. 2. The GCYMS profiles of the same samples are reported in Figs. 3 and 4.
Metals investigated were Fe and Zn because of their relatively high amount
(ranging from 20 to 188 ppm), the results about FelZn amount in heroin samples
are reported in Figs. 5 and 6. By the outlined analytical sequence, we obtained

LNn4ean4-e.2n-J
Fig. 6. FelZn relative amount in heroin samples (for analytical conditions see the text).

56

information based on the identification of compounds carried over from the

opium raw material or resulting by the acetylation process. So, other than major
opiates and related compounds, we also determined traces of volatile substances
employed during the extraction, acetylation and crystallization steps. Moreover,
fluorescent chromatographic patterns, mainly related to thebaol, acetylthebaol
and phenanthrenic derivatives of thebaine, were produced during the synthesis
of heroin. Additional information including metals, diluents and adulterants profiles was useful to attribute or exclude common sources of individual specimens.
As a matter of fact, the chemical composition elucidated by the proposed
analytical sequence on 33 heroin samples (Table 2) gave us sufficient information
to detect similarity between samples 5,32; 333; 11,28; 25,26. On the contrary,
we can exclude common sources between samples 19, 31; 2, 33; 14, 16; which
demonstrated the same heroin content or opiate profile but showed some differences of volatiles, and/or metals.
References

7
8
9
10

11
12
13
14
15

K. Narayaneswami, Parameters for determining the origin of illicit heroin samples. Bull. NUT-C.,
XXXVII (1985) 49-62.
P.J. ONeil et al., Illicitly imported heroin products: some physical and chemical features indicative of their origin. J. Ftn-emic Sci., 29 (1984) 889-902.
V. Rajananda et al., An evaluation of TLC system for opiate analysis. Bull. Narc., XXXVII
(1985) 35-47.
H. Huiser, Analytical studies on illicit heroin. I. The occurrence of O-monoacetylmorphine. J.
Forensic Sci., 28 (1983) 32-39.
H. Neumann, Analysis of opium and crude morphine samples by capillary as chromatography;
comparison of impurity profiles. J. Chmmatogr., 315 (1984) 404-411.
J.M. Moore, Rapid and sensitive gas chromatographic qua&&ion
of morphine, codeine and
O%cetylmorphine
in illicit heroin using an electron capture detector. J. Chmmdogr., 147
(1978) 327-336.
M.D. Miller, The determination of excipient sugar diluents in illicit preparations containing
heroin by gas chromatography. J. Forensic Sci., (1971) 150-163.
E. Kaa and K. Bent, Impurities, adulterants and diluents of illicit heroin in Denmark (Jutland
and Funen). Fwti
Sci. Id., 31 (1986) 195-210.
M. Chiarotti and N. Fucci, Analysis of volatile compounds in heroin samples. Forensic Sci. Znt.,
37 (1988) 47-53.
J.M. Moore et al., Determination of manufacturing impurities in heroin capillary CC with electron capture detection after derivatiaation with heptafluorobutyric anhydride. And. Chem., 56
(1984) 642-649.
C.E. Cook and D.R. Brine, Pyrolysis products of heroin. J. Forensic Sci., 30 (1985) 251-261.
A.C. Allen et al., Illicit heroin manufacturing byproducts. And. Chin., 56 (1984) 2940-2947.
J. Org. Chew.., 19 (1954) 1409-1415.
L.H. Welsh, 03-monoacetylmorphine.
H. Huiser, Analytical studies on illicit heroin. II. Comparison of samples. J. Forensic Sci., 28(l)
(1983) 40-48.
Heroin manufacture use of ethylidene diacetate. Microgram, 20 (1987) 195.