Forensic Science International

92 (1998) 125–136

Capillary electrophoresis of illicit drug seizures

Ira S. Lurie*
U.S. Drug Enforcement Administration, Special Testing and Research Laboratory, 7704 Old Springhouse
Road, McLean, VA 22102 -3494, USA

Received 29 August 1997; accepted 23 September 1997

Abstract

Capillary electrophoresis (CE) is well suited for the analysis of illicit drug seizures. CE
techniques can separate a wide variety of solutes, including compounds that are highly polar,
thermally labile and / or nonvolatile, with high efficiency and selectivity. Micellar electrokinetic
chromatography (MECC), which can separate basic, acidic and neutral solutes, is an excellent
technique for general drug screening and the analysis of heroin, opium, cocaine, LSD, anabolic
steroids, and the diastereomers formed from derivatized optical isomers of phenethylamines.
Capillary zone electrophoresis (CZE) has been used for the analysis of small ions (cations and
anions) in illicit heroin. Cyclodextrin (CD)-modified CZE (i.e. CZE using run buffers containing
either charged and / or neutral CDs) has proven successful for both chiral and achiral analyses.
Chiral applications include khat, phenethylamines, cocaine, propoxyphene and methorphan, while
achiral applications include the analysis of naturally occurring impurities present in illicit cocaine,
including truxilline diastereomers and hydroxycocaine endo and exo diastereomers. Capillary
electrochromatography (CEC), which is a hybrid between high performance liquid chromatog-
raphy (HPLC) and CE, also shows great promise for forensic drug analysis. Published by Elsevier
Science Ireland Ltd.

Keywords: Capillary electrophoresis; Illicit drug seizures; Micellar electrokinetic capillary

chromatography; Capillary zone electrophoresis; Cyclodextrins

1. Introduction
In addition to the controlled substance(s), drug exhibits typically contain various
adulterants, diluents and manufacturing by-products and impurities. The comprehensive

*Corresponding author: Tel.: 1-703-285-2583; fax: 1-703-285-2198.

0379-0738 / 98 / $19.00 Published by Elsevier Science Ireland Ltd.

PII S0379-0738( 98 )00012-7
126 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136

analysis of these solutes is important for both legal and intelligence requirements [1].
Capillary electrophoresis (CE) is useful for the analysis of the wide variety of solutes
found in illicit drug seizures, especially for those compounds which are otherwise
difficult to analyze via gas chromatography (GC) and high performance liquid
chromatography (HPLC). GC can be problematic for the analysis of nonvolatile,
thermally labile and highly polar drugs, while HPLC often lacks sufficient resolving
power for complex exhibits. In addition, although enantiomers of forensic interest can be
resolved by either GC and HPLC, derivatization and / or the use of expensive, specialized
columns are usually required [2]. CE, which has been commercially available for over 8

Table 1
Migration times of compounds of forensic interest relative to heroin
Compound Relative migration time
Benzoylecgonine 0.38
Psilocybin 0.46
Morphine 0.51
Dilaudid 0.54
Phenobarbital 0.58
Psilocin 0.62
O 6 -Monoacetymorphine 0.66
O 3 -Monoacetylmorphine 0.72
Codeine 0.74
Methaqualone 0.78
Mescaline 0.92
LSD 0.93
LAMPA 1.00
Heroin 1.00 (8.91 min)
Acetylcodeine 1.14
Papaverine 1.24
MDA 1.27
Amphetamine 1.28
Librium 1.34
Cocaine 1.49
MDMA 1.55
Methamphetamine 1.56
Phentermine 1.56
Noscapine 1.69
cis-Cinnamoylcocaine 1.76
Lorazepam 1.91
Diazepam 2.21
trans-Cinnamoylcocaine 2.24
Fentanyl 2.36
Flurazepam 3.07
PCP 3.23
Pyrene a 3.72
Cannabidiol 3.90
Cannabinol 4.25
D9 -THC 4.25
a
Added as a potential t m c marker. See text for discussion.
Reproduced from Reference [3].
 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136 127

years, offers high efficiency, high selectivity, and economy of operation, and therefore
offers great potential to the forensic chemist. The economy of operation arises from the
low flow rates (nl / minute) and capillary costs (approximately ten dollars).
This laboratory has been involved with CE for over seven years. CE has been applied
for both general screening [3] and in-depth analyses of seized drugs, including heroin
[3–6], cocaine [2,3,7,8], opium [9], khat [2], anabolic steroids [10], propoxyphene [2],
methorphan [11], LSD [12], and the amphetamines [2,13]. Our earlier work in this area
has been previously reviewed [11]. This paper reviews our efforts in micellar
electrokinetic capillary chromatography (MECC), capillary zone electrophoresis (CZE),
cyclodextrin (CD)-modified CZE, and capillary electrochromatography (CEC).

2. MECC

MECC using micelles such as sodium dodecyl sulfate (SDS) is an excellent technique
for forensic drug analysis, as it allows for the analysis of acidic, neutral and basic
compounds, including structurally similar solutes. In addition, since all solutes must
elute by the time of the micelle (t m c ), the technique is well suited for general drug
screening [3]. The migration times for various compounds, including opium alkaloids,
amphetamines, hallucinogens, barbiturates, benzodiazepines and cannabinoids, are
shown in Table 1; an electropherogram of selected solutes is displayed in Fig. 1. By
shortening the capillary and using a lower concentration of micelle, run times of under

Fig. 1. MECC forensic drug screen. Conditions: capillary, 47 cm (25 cm length to detector)350 mm id;
voltage, 20 kV; temperature, 40 degrees centigrade; buffer, 85 mM SDS / 8.5 mM phosphate / 8.5 mM
borate / 15% acetonitrile, pH 8.5; detector wavelength, 210 nm, sample concentration, 250 mg of each drug.
Key: (a) psilocybin, (b) morphine, (c) phenobarbital, (d) psilocin, (e) codeine, (f) methaqualone, (g) LSD, (h)
heroin, (i) amphetamine, (j) librium, (k) cocaine, (l) methamphetamine, (m) lorazepam, (n) diazepam, (o)
fentanyl, (p) PCP, (q) cannabidiol, (r) D9 -THC. Reproduced from Ref. [3].
128 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136

10 minutes are easily obtainable. In addition, greater specificity can be obtained by using
photodiode array (PDA) or rapid scan UV detection, which allows for the acquisition of
UV spectra. Depending on the software package used, automatic library searches can be
performed. Many drug exhibits (e.g. heroin) contain mixtures of basic, acidic and neutral
solutes. As shown in Fig. 2, MECC allows for the analysis of a complex heroin sample
in under three minutes [4]. O6-acetylmorphine separates from O3-acetylmorphine and
acetylcodeine, solutes which can be difficult to separate via CZE. It is interesting to note
that the same separation via HPLC requires approximately one hour, including gradient
equilibration time [3]. Good quantitative data was obtained for solutes such as heroin,
noscapine and methaqualone [4].
In a similar vein, MECC is applicable to the analyses of opium [9], cocaine [3] and
LSD [12]. CZE is not amenable to compounds with similar charge-to-size ratios, such as

Fig. 2. An electropherogram of an illicit heroin sample. Conditions: capillary, 27 cm (20 cm length to

detector)350 mm id; voltage, 20 kV; temperature, 30 degrees centigrade; buffer, 45 mM SDS / 8.5 mM
phosphate / 8.5 mM borate / 15% acetonitrile, pH 8.5; detector wavelength, 214 nm. Key: (A) phenacetin, (B)
caffeine, (C) morphine, (D) O6-monoacetylmorphine, (E) codeine, (F) methaqualone, (G) phenobarbital, (H)
NPPB (internal standard), (I) heroin, (J) acetylcodeine, (K) papaverine (L) thebaine, (M) noscapine.
Reproduced from Ref. [4].
 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136 129

Fig. 3. MECC electropherogram of opium sample. A 57 cm (50 cm length to detector)350 mm id capillary at

30 degrees centigrade was used with a voltage of 21 kV. The run buffer contained 9.0 mM phosphate, 9.0 mM
borate, 45.0 mM SDS, 6.9 mM b -CD-SBE(IV), and 10% acetonitrile. Key: (a) morphine, (b) codeine, (c)
papaverine, (d) thebaine, (e) noscapine. Reproduced from Ref. [9].

morphine, codeine and thebaine in opium [14]. An electropherogram of Southwest Asian

opium is shown in Fig. 3. Similarly, cis- and trans-cinnamoyl cocaine, two major
impurities commonly present in illicit cocaine, are easily resolved (same conditions as
Fig. 2), and LSD, lysergic acid methyl propyl amide (LAMPA) and iso-LSD are
resolved in under five minutes (same conditions as Fig. 2).
In a related application, MECC provides an excellent separation of the highly complex
mixture of neutral and acidic impurities present in heroin [5] (see Fig. 3 for CE
conditions). Using injection techniques, such as stacking with UV detection and / or laser
induced fluorescence (LIF) detection, allows for the ultra trace level analysis of these
solutes.
Anabolic steroids and their esters can be separated in a single run (see Fig. 4),
analogous to the methodologies for these solutes using temperature programmed
capillary GC and gradient elution HPLC [10]. All three techniques give nearly
equivalent resolution. However, although capillary GC exhibits higher peak capacity
than MECC, the latter technique is more selective for anabolic steroids and their esters,
and also precludes problems with thermal decomposition in heated GC injection ports.
Due to the high amount of acetonitrile present in the MECC run buffer (required for
solubility considerations), less than optimum k9 values are obtained for these solutes. All
three techniques have been shown to be orthogonal, and MECC gives unique chromato-
130 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136

Fig. 4. MECC electropherogram of anabolic steroid mixture. A 72 cm (50 cm length to detector)350 mm id

capillary at 30 degrees centigrade was used with a voltage of 25 kV. The run buffer was acetonitrile / sodium
dodecyl sulfate (SDS) solution 2:3; the SDS solution contained 75 mM SDS in 10 mM phosphate / 10 mM
borate buffer at pH 9.0. Key: (a) boldenone, (b) methandrostenolone, (c) testosterone, (d) methyltestosterone;
(e) methadriol, (f) stanolone, (g) boldenone acetate, (h) danazol, (i) testosterone acetate, (j) nandrolone
propionate, (k) methandriol-3-acetate, (l) testosterone isobutyrate, (m) testosterone cypionate, (o) testosterone
undecanoate. Reproduced from Ref. [10].

graphic data. MECC has been successfully applied for the quantitative determination of
various anabolic steroid preparations, including tablets, aqueous suspensions and oils.
Finally, MECC has been applied for the separation of 2,3,4,6-tetra-O-acetyl-b -D-
glucopyranosyl isothiocyanate (GITC) derivatives of enantiomers of norephedrine,
norpseudoephedrine, ephedrine, pseudoephedrine, amphetamine and methamphetamine
[13]. The electropherograms of a standard mixture of all of these solutes, and an illicit
methamphetamine sample, are shown in Fig. 5.

3. CZE

CZE has been used for the analysis of small to moderate sized ions in heroin exhibits
[6]. Cations, including ammonium, potassium, sodium, calcium and magnesium, are
separated in under three minutes using CZE with indirect UV detection (see Fig. 6).
Anions, including chloride, sulfate, citrate, tartrate, phosphate and acetate are separated
in under six minutes, also using CZE with indirect UV detection (see Fig. 7). The
methodology is capable of determining ions down to the 10 2 3 % level relative to heroin,
using lithium nitrate as an internal standard.
 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136 131

Fig. 5. MECC separation of phenethylamine-GITC derivatives. Standard mixture (top); seized exhibit
(bottom). Conditions: capillary, 48 cm350 mm id (length to detector 26 cm); voltage, 20 kV; temperature, 30
degrees centigrade; buffer, 80 mM SDS / 8.0 mM borate / 8.0 mM phosphate / 20% methanol, pH 9.0 Key: (a)
(1)-norpseudoephedrine, (b) (2)-ephedrine, (c) (2)-norephedrine, (d) (2)-norpseudoephedrine, (e) (1)-
ephedrine, (f) (1)-pseudoephedrine, (g) (1)-norephedrine, (h) (2)-pseudoephedrine, (i) (2)-metham-
phetamine, (j) (2)-amphetamine, (k) (1)-methamphetamine, (l) (1)-amphetamine. Reproduced from Ref.
[11].

4. CD-modified CZE

Run buffers containing either a neutral CD, a charged CD, or mixtures of neutral and
charged CDs, have proven invaluable for forensic drug applications, including the
analysis of enantiomers [2] and diastereomers [7,8]. This mode of CE analysis is
particularly attractive because derivatization is not required.
The neutral CD heptakis[2,6-di-O-methyl]-b -cyclodextrin] (DM-b -CD) has been
used for the analysis of khat [2]. (2)-Cathinone and (1)-norpseudoephedrine (cathine),
the major physiologically active compounds present in khat, are both controlled by
international statutes. CD-CZE allows the separation of most of the enantiomers of the
phenylalkylamines present in khat, including cathinone, norpseudoephedrine, norephe-
drine, merucathinone, merucathine and pseudomerucathine (see Fig. 8). The comigration
of (2)-merucathine and (2)- pseudomerucathine evident in Fig. 8 is not a concern,
because the former solute is not believed to be present in khat.
Mixtures of DM-b -CD and the anionic cyclodextrin b -CD sulfobutyl ether IV
( b -CD-SBE(IV)) have been used for the separation of enantiomers of chiral drugs of
forensic interest, including primary, secondary and tertiary amines [2]. This system
allows the simultaneous resolution of the (1,2)-amphetamine and its precursors (1,2)-
norephedrine and (1,2)-norpseudoephedrine, and has also been used for the simulta-
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Fig. 6. Electropherogram of standard mixture of cations. Conditions: capillary, 48 cm (26 cm effective

length)350 mm id; injection, vacuum, 4.0 s; voltage 20 kV; temperature, 30 degrees centigrade; buffer, Ion
Phor Cation CU (Dionex); detector wavelength, 210 nm. Key: (a) ammonium (2 mg / ml), (b) potassium (2
mg / ml), (c) sodium (5 mg / ml), (d) calcium (3 mg / ml), (e) magnesium (1 mg / ml), (f) lithium (1 mg / ml)
(internal standard). Reproduced from Ref. [6].

neous resolution of (1,2)-methcathinone, (1,2)-methamphetamine, and their pre-

cursors (1,2)-ephedrine and (1,2)-pseudoephedrine (see Fig. 9). A similar system has
been used to resolve the enantiomers of the tertiary amines cocaine [2], propoxyphene
[2] and methorphan [11].
Eight of the truxilline diastereomers and several other high molecular weight
impurities found in illicit cocaine have been analyzed using b -CD-SBE(IV) [7] (see Fig.

Fig. 7. Electropherogram of standard mixture of anions. Conditions: capillary, 72 cm (50 cm effective

length)350 mm id; injection, vacuum, 4.0 s; voltage, 230 kV; temperature, 30 degrees centigrade; buffer,
Anitron (Perkin–Elmer); detector wavelength 230 nm. Key: (a) chloride (9 mg / ml), (b) sulfate (8 mg / ml), (c)
nitrate (11 mg / ml) (internal standard), (d) citrate (10 mg / ml), (e) tartrate (10 mg / ml), (f) phosphate (11
mg / ml), (g) carbonate (7.0 mg / ml), (h) acetate (10 mg / ml).
 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136 133

Fig. 8. Electropherogram of khat leaf standards. Conditions: Capillary 82 cm350 mm id (length to detector 60
cm); voltage, 30 kV; temperature, 30 degrees centigrade; buffer 4.5 mM DM-b -CD/ 22.5 mM tris-H 3 PO 4 / 10%
methanol, pH 2.45; detector wavelength, 210 nm. Key: (a) (1)-cathinone, (b) (2)-cathinone, (c) (2)-
norpseudoephedrine, (d) (2)-norephedrine, (e) (1)-norephedrine, (f) (1)-norpseudoephedrine (cathine), (g)
(2)-merucathinone, (h) (1)-merucathinone, (i) (1)-merucathine, (j) (1)-pseudomerucathine, (k) (2)-
merucathine, (1) (2)-pseudomerucathine. Reproduced from Ref. [2].

Fig. 9. Electropherograms of secondary amine standards. Conditions 1.2% methanol, 98.8% 5 mM DM-b -CD
1 mM b -CD-SBE(IV), 25 mM tris-H 3 PO 4 , pH 2.45. Capillary, 82 cm (60 cm effective length)350 mm id;
voltage, 30 kV; detection, UV, 210 nm; temperature, 30 degrees centigrade. Key: (a) (1)methcathinone, (b)
(2)methcathinone, (c) (2)-pseudoephedrine, (d) (2)-ephedrine, (e) (1)-ephedrine, (f) (1)-pseudoephedrine,
(g) (2)-methamphetamine, (h) (1)-methamphetamine.
134 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136

10). In addition, a mixture of 6- and 7-hydroxycocaine endo, exo isomers are separated
using a b -CD-SBE(IV) / DM-b -CD mixture very similar to that used for the separation
of enantiomers of tertiary amines (no methanol present) [2,8]. It is of interest to note that
when using HPLC, these cocaine impurities were separated only by the operationally
difficult normal phase technique [15].

Fig. 10. Electropherogram of an illicit cocaine HCl HPLC size exclusion extract. A 58 cm (50 cm length to
detector)350 mm id capillary at 30 degrees centigrade was used with a voltage of 27.5 kV. UV detection at 210
nm. The run buffer contained 10% methanol and 90% (10 mM b -CD-SBE(IV), 50 mM phosphate, pH 8.6).
Reproduced from Ref. [7].
 I.S. Lurie / Forensic Science International 92 (1998) 125 – 136 135

5. CEC

Preliminary work has begun in our laboratory using capillary HPLC columns with
electrically driven flow for the analysis of hydrophobic solutes, e.g. anabolic steroids and
cannabinoids. Capillary electrochromatography (CEC) incorporates the high efficiency
obtained with CE with the high selectivity obtained with HPLC. Since pressure drop for
CEC is not a problem (as in HPLC), longer columns can be used with very small particle
sizes (e.g. 0.5 mm) to generate plate counts numbering in the hundreds of thousands.
Thus, if gradient elution is used, CEC has the capability to generate peak capacities
approaching temperature programmed capillary GC [16]. CEC also provides higher
sample capacities versus conventional CE.

6. Conclusion

Capillary electrophoresis techniques, including MECC, CZE and CD-CZE, have

shown great utility for the analyses of illicit drug seizures. These high resolution
techniques are applicable to a large variety of solutes present in forensic drug exhibits,
including compounds that are nonvolatile, thermally labile and highly polar. In the near
future, CEC shows great promise for the analyses of illicit drug seizures.

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