Original Article Ann Clin Biochem 2000; 37: 690±700

Analysis of urine for drugs of abuse using mixed-mode solid-phase

extraction and gas chromatography±mass spectrometry
Susan Paterson1, Rosa Cordero1, Scott McCulloch2 and Philip Houldsworth2
From the 1Toxicology Unit, Imperial College School of Medicine, St Dunstan’s Road, London W6 8RP,
and 2Tackler Analytical, 18 Green Road, Whetstone, London N20 0QT, UK

SUMMARY. A method for the simultaneous analysis of urine for the major drugs of
abuse is described. The analytical procedure uses solid-phase extraction (SPE), gas
chromatography±mass spectrometry (GC±MS) and a semi-automated identi®cation
process. It allows simultaneous extraction, derivatization and analysis of acidic,
neutral and basic drugs from urine. Urine samples were subjected to enzymatic
hydrolysis followed by SPE using Bakerbond narc-2 columns. The eluant was
selectively derivatized with N-methyl-bis-tri¯uoroacetamide (MBTFA) and N-
methyl-N-trimethylsilyltri¯uoroacetamide+1% trimethylchlorosilane. Analysis was
performed using a GC±MS system operating in full scan mode. A simple macro
programme was written to enhance the mass spectra identi®cation capabilities of the
MS software by producing extracted ion chromatograms (EIC) for the drugs of
interest. Once a suspect compound was indicated by EIC, the mass spectrum of the
compound was searched manually against reference libraries for positive
identi®cation and the retention time checked against that of the standard. This
procedure has increased both the amount and the reliability of information given to
clinicians without increasing the cost per sample. The system has been in routine
operation for 24 months, processing up to 40 urine samples per day, with a usual
turn-around time of 48 h.

INTRODUCTION from drug treatment centres. However, legal

The Toxicology Unit was approached to provide
a urine screening service for the speci®c target
population of clients attending drug-dependency
clinics. The method described here is designed
speci®cally for this type of work. The require-
ments for the provision of an ef®cient and
reliable service for screening drugs of abuse
include rapid turn-around times, good commu-
nications between requesting clinician and the
laboratory, and participation in quality assess-
ment schemes. In addition, there is a need to
con®rm positive results from screening tests.1 In
Britain, most laboratories performing workplace
drug testing follow guidelines set by the
Substance Abuse and Mental Health Services
Administration, which state that any positive
screening results must be con®rmed using gas
chromatography±mass spectrometry (GC±MS).2
This is not always deemed necessary for samples
Correspondence: Dr S Paterson.
E-mail: s.paterson@ic.ac.uk
implications can follow from any result. The
main reason why a similar analytical procedure
is not used for both drug treatment centre
screening and workplace testing is due to the
number of samples testing positive for drugs.
For workplace drug testing, immunoassay is
used for screening and most samples test
negative. This means that there are very few
samples to be con®rmed using GC±MS, a
technique that is expensive and requires an
operator with a high degree of expertise. In
drug treatment centre screening, the vast
majority of samples test positive and many are
positive for more than one drug. It is therefore
not realistic to treat these samples with the same
approach. For drug treatment centre screening,
a whole range of other methods is generally
employed ± such as immunoassays, thin-layer
chromatography (TLC) and gas chromatogra-
phy (GC) ± which are less expensive and do not
require such a highly skilled operator. In order
to analyse samples from drug treatment centres

690

using GC±MS, a method is needed that allows
extraction of the whole range of drugs of abuse
simultaneously. It would then follow that only
one injection per sample onto the GC±MS
system would be necessary. This would then
make the use of GC±MS analysis of drug
treatment samples a realistic proposition and
would provide a single assay per sample that
would give an unequivocal evaluation of a
patient’s drug status.
With the advent of mixed-mode solid-phase
extraction (SPE) columns, it has become
possible to isolate acidic, neutral and basic
drugs using a single column.3±5 The method
presented here, which was developed in-house,
uses a mixed-mode SPE extraction column from
which a whole range of drugs of abuse are
extracted simultaneously. The extracted drugs
are then derivatized with N-methyl-N-trimethyl-
silyltri¯uoroacetamide+1% trimethylchlorosi-
lane (MSTFA+1% TMCS) and N-methyl-bis-
tri¯uoroacetamide (MBTFA). The use of a
three-stage derivatization procedure utilizing
MBTFA and MSTFA+TMCS has been pre-
viously described.4 The initial addition of
MBTFA prevented the loss of volatile primary
and secondary amines by derivatization. It was
shown that the addition of the derivatization
reagent to the eluent prior to drying down did
not cause problems in the procedure.
A GC±MS system in scan mode is used to
detect and identify the drugs. Using Hewlett-
Packard ChemStation software, at least 61 drugs
and metabolites are routinely looked for in all
samples. These include all the commonly abused
drugs, plus a whole range of other drugs
including antidepressants and certain antipsy-
chotics that are prescribed to these patients. The
samples are incubated prior to extraction with b-
glucuronidase to enhance the detection of
benzodiazepines6 and morphine.7
MATERIALS AND METHODS
Materials
All chemicals were of analyticalreagentgrade and
were obtained from Merck (Poole, UK). Deuter-
ated methylenedioxyamphetamine (MDA-d5)
was obtained from Promochem (Welwyn Garden
City, UK). All drug standards, b-glucuronidase
type H-2 (crude solution from Helix pomatia,
132 500 units/mL; EC 3.2.1.31), MBTFA and
ammonium hydroxide ACS reagent (28´0±30´0%
NH3) were obtained from Sigma-Aldrich (Poole,
UK). Drugs of abuse urine control module
Analysis of urine for drugs of abuse 691
(CON-DAU) level 3 was obtained from DPC
(Llanberis, UK). Bakerbond narc-2 jumbo 3-mL
(125-mg) extraction cartridges were obtained
from Mallinckrodt Baker (Milton Keynes, UK).
The Spe-ed wiz vacuum manifold was obtained
from Applied Separations (Allentown, USA).
The Techne sample concentrator was obtained
from Merck (Poole, UK). MSTFA+1% TMCS
was obtained from Pierce & Warriner (Chester,
UK).
Solutions
Acetate buffer, 1´0 mol/L pH 5´0, was prepared
by dissolving 42´9 g of sodium acetate trihydrate
in 400 mL of distilled water; 10´4 mL of glacial
acetic acid were added and the solution was
diluted to 500 mL with distilled water. The pH
was adjusted to 5´0 with either 1´0 mol/L
potassium hydroxide or 1´0 mol/L hydrochloric
acid. Phosphate buffer, 0´1 mol/L pH 6´0, was
prepared by dissolving 13´61 g of anhydrous
potassium dihydrogen phosphate in 900 mL of
distilled water. The pH was adjusted to 6´0 with
1´0 mol/L potassium hydroxide and the ®nal
solution was made up to 1´0 L with distilled
water. Dilute acetic acid, 10 mmol/L pH 3´3,
was prepared by diluting 574 mL of glacial acetic
acid to 1´0 L with distilled water.
Preparation of internal standard solution
MDA-d5 was used as an internal standard; 1 mL
of a 100 mg/mL solution of MDA-d5 in
methanol was diluted to 150 mL with acetate
buffer, 1´0 mol/L pH 5´0. MDA-d5 is used as the
internal standard because, like all ampheta-
mines, it is volatile. In addition, it is derivatized
by both of the derivatizing agents used. It
therefore monitors all these stages of the
analysis.
Sample pre-treatment
To 4 mL of urine, 1´5 mL of internal standard
solution and 50 mL (6600 units) of b-glucuroni-
dase were added. The samples were capped and
incubated overnight at 608C. After incubation
the samples were allowed to cool. The pH of
each sample was then adjusted to 8´0 by the
addition of 445 mL of 1´0 mol/L potassium
hydroxide solution and adjusted further if
necessary. The samples were centrifuged at
3000 rpm for 10 min.
Extraction procedure
The extraction was performed using Bakerbond
narc-2 columns installed in a Spe-ed wiz vacuum
Ann Clin Biochem 2000: 37
692 Paterson et al.
manifold. The procedure consists of the follow-
ing steps:
1. The columns were conditioned with 2´0 mL
of methanol followed by 2´0 mL of distilled
water followed by 2´0 mL of 0´1 mol/L
phosphate buffer pH 6´0.
2. The supernatant from the pre-treated sam-
ples was added to the column and pulled
through at a rate of less than 5 mL/min.
3. The column was washed with 1 mL of
distilled water followed by 0´250 mL of
10 mmol/L acetic acid at pH 3´0.
4. The samples were eluted from the columns
with 3´0 mL of chloroform±isopropyl alco-
hol (80:20, v/v), followed by 3´0 mL of
chloroform±isopropyl alcohol±ammonium
hydroxide ACS reagent (80:20:3, v/v). Both
eluates were added to the same tube.
Derivitization procedure
1. To each eluent, 25 mL of MBTFA were
added. After mixing, the solutions were
evaporated under nitrogen at 708C to ap-
proximately 1´0 mL using a Techne sample
concentrator.
2. The remaining solution was transferred to a
microvial. The solutions were then evapo-
rated to dryness under nitrogen at 708C.
3. The vials were removed from the heating
block and 50 mL of MSTFA±TMCS were
added to each. The vials were capped and
incubated at 708C for 60 min.
4. Then 25 mL of MBTFA were added to each
vial and the vials were incubated for a
further 30 min at 708C.
5. A 1-mL aliquot was injected onto the GC±
MS system.
Instrumental analysis
A Hewlett-Packard (UK) model 5973 mass
selective detector with a 6890 series gas chroma-
tograph ®tted with a split/splitless injection port,
an HP G1513A automatic sampler and the HP
Enhanced Productivity MS Chemstation soft-
ware were used. The analytical column was an
HP-5 MS (30 m60´25 mm i.d., 0´25 mm ®lm
thickness), ®tted with an HP Retention Gap
(uncoated, deactivated) (1 m60´25 mm i.d., 0
mm ®lm thickness). Temperature conditions were
as follows: initial temperature 808C for 2 min;
increased at 168C/min to 2908C; held for
5´37 min; giving a total run time of 20´50 min.
The ¯ow of the carrier gas (helium) was
maintained at 1´0 mL/min in constant ¯ow
Ann Clin Biochem 2000: 37
mode. The mass spectrometer was operated with
an electron energy of 70 eV and a mass-to-
charge ratio scan range of 50±550 amu. The ion
source and the quadrupole temperatures were
maintained at 2308C and 1508C, respectively.
The GC±MS system was programmed to per-
form a 1-mL splitless injection.
Data acquisition was performed using an HP
Vectra XM series 4 computer, running HP
G1701AA Enhanced Chemstation software
(version G1701AA, revision A.03.00). A macro
was written in-house for the software, which
performed target compound analysis on each
sample for the 61 speci®ed drugs. Once isolated
from the total ion chromatogram, positive
identi®cation was provided by three libraries: a
library developed in-house, containing many
drug tri¯uoroacetamide (TFA) and trimethylsi-
lyl (TMS) derivatives (274 compounds); the
P¯eger Maurer Webber library (4367 com-
pounds); and the Wiley 275 library (275 821
compounds).
Control material
A control was run with each batch of samples to
check retention times and performance of the
assay. The control material was diluted with
blank urine to give the following concentrations
in mmol/L (mg/L): d-amphetamine 9´25 (1´25),
benzoylecgonine 1´3 (0´375), codeine 2´5 (0´75),
methadone 0´8 (0´25), d-methamphetamine 8´4
(1´25), morphine 0´26 (0´075), morphine-3-glu-
curonide 2´85 (1´325), oxazepam 0´875 (0´25), d-
propoxyphene 0´55 (0´188), secobarbital 2´1
(0´5), tetrahydrocannabinoicacid 0´183 (0´0625).
Recoveries
Solutions of the drugs of abuse listed in Table 3
were prepared and added to blank urine to give
®nal concentrations in urine of 2´0 mg/L for all
drugs except 9-THC-11-oic acid and MDA-d5.
As 9-THC-11-oic acid is usually present in
much lower concentrations, this drug was added
to blank urine to give a ®nal concentration of
0´10 mg/L. Aliquots (4 mL) of these spiked
urine solutions were extracted in the same
manner as the samples. An external standard,
50 mL of a 1´34 mg/L solution of amitriptyline
in methanol, was added. The extracts were then
derivatized and run on the GC±MS system as
normal. Solutions of the same drugs were
prepared in chloroform±isopropanol (80:20,
v/v) to give the same ®nal concentrations as in
blank urine. Aliquots (4 mL) of these solutions
were taken and to these were added 50 mL of the
 Analysis of urine for drugs of abuse

FIGURE 1. Total ion chromatogram from a control sample. 1=amphetamine±TFA; 2=methamphetamine±TFA; 3=methylenedioxyamphetamine-d5±TFA/TMS; 4=
secobarbital±TMS; 5=phencyclidine; 6=methadone; 7=benzoylecgonine±TMS; 8=oxazepam±TMS; 9=codeine±TMS; 10=morphine±TMS; 11= 9-THC-11-oic

Ann Clin Biochem 2000: 37

693

acid±TMS. TFA=tri¯oroacetamide; TMS=trimethylsilyl.

 694

Ann Clin Biochem 2000: 37

Paterson et al.

FIGURE 2. Total ion chromatogram from a patient’s sample. 1=ecgonine methyl ester±TMS; 2=methylenedioxyamphetamine-d5±TFA/TMS; 3=methadone; 4=2-ethyl-5-
methyl-3,3-diphenyl-1-pyrroline (methadone metabolite); 5=methadone±TMS; 6=cocaine; 7=benzoylecgonine±TMS; 8=desmethyldiazepam±TMS; 9=oxazepam±TMS;
10=codeine±TMS; 11=morphine±TMS; 12=temazepam±TMS. TFA=tri¯uoracetamide; TMS=trimethylsilyl.
 Analysis of urine for drugs of abuse 695

same external standard solution. The extracts the extraction column is pre-conditioned to pH
were then derivatized and run on the GC±MS 6´0, it was found that the optimum pH for
system as normal. The ratio of the extracted retention of the whole range of drugs of abuse
drug to external standard was compared to the on the extraction column is 8´0. As endogenous
ratio of non-extracted drug to external standard. compounds found in urine are water-soluble,
By comparing these ratios the percentage they were removed from the column by washing
recovery was calculated. with water. After washing, acetic acid was added
to the column adjusting the pH to 3´0. At this
RESULTS pH, acid and neutral drugs are in the unionized
form and therefore elute in chloroform±isopro-
Figures 1 and 2 show total ion chromatograms
pyl alcohol. The basic drugs were then eluted
from the control (Fig. 1) and from a patient’s
with chloroform±isopropyl alcohol made basic
sample (Fig. 2). It can be seen that the
by the addition of ammonia solution. Water was
chromatograms are relatively clean. A simple
found to provide the optimal type and volume of
macro programme was written in-house to
wash for maximum compound retention by the
produce extracted ion chromatograms (EIC)
extraction column, while removing the interfer-
for the detection of the 61 compounds that each
ing endogenous compounds contained in the
sample is routinely screened for. The mass
urine. The inclusion of methanol in the wash
spectroscopist only views the total ion chroma-
caused a decrease in the recovery of the drugs.
togram to see if there are any unusual peaks
Table 3 shows the results of the recovery
present. Table 1 shows the retention times and
experiments for the major drugs of abuse.
mass spectral data used to perform target
Recoveries for 9-THC-11-oic acid, a strong
compound analysis and produce the EIC. A
acid, and for temazepam and oxazepam, neutral
print-out is issued for each sample showing a
drugs, are relatively low but they are acceptable.
chromatogram of the three selected m/z ions for
The eluent was selectively derivatized with
each compound. If the EIC indicates the
MBTFA, which formed tri¯uoroacetamide
presence of a particular compound by showing
(TFA) derivatives of primary and secondary
that the three ions are present and are present in
amines, and MSTFA+1% TMCS, which
approximately the correct ratio, then the spectra
formed trimethylsilyl (TMS) derivatives of
and retention time are matched manually. The
hydroxyl, acidic and phenolic groups. If the
internal standard is not used for quantitation
evaporation step was performed without the
work; it is added to check that the extraction
addition of MBTFA to the eluent it resulted in
and derivatization procedures have been com-
the loss of low molecular weight stimulants, i.e.
pleted successfully. If the internal standard is
amphetamines. Solans et al.4 observed that, after
shown not to be present in the EIC, then the
the addition of MSTFA+1% TMCS to the
sample is re-run. The reference libraries used
dried residue at 808C for 1 h, the formation of
include the P¯eger Maurer Webber library, the
TMS and TFA derivatives was distorted because
Wiley 275 library and an in-house library. The
the TMS took the place of some TFA in some
latter has been obtained by recording the spectra
amine groups. It was therefore necessary to add
of derivatized standards of the various com-
a further 25 mL of MBTFA to complete the
pounds analysed by GC±MS. Figure 3 shows the
derivatization of the primary and secondary
ion chromatogram for morphine±TMS from a
amines.
patient’s sample compared with that of mor-
Some drugs give more than one derivatization
phine±TMS from the in-house reference library.
product. These include methadone, which has
Looking at the EICs alone will show which
both an underivatized and a TMS derivative
drugs are absent, but to be certain of the
peak, and MDA, which has both a TFA and a
presence of those indicated by the EIC it is
TFA/TMS derivative peak. For each of these
always necessary for a mass spectroscopist to
drugs, the major peak was used to estimate the
match the spectra manually.
limit of detection and calculate percentage
recovery, namely the TMS derivative peak for
DISCUSSION
methadone and the TFA/TMS derivative peak
The Bakerbond narc-2 column contains C18 and for MDA.
cationic exchange adsorbents (aromatic sulpho- The method is used to produce qualitative
nic acid) and the drugs are retained by a mixture results only. To be reported as positive, the drug
of ionic and hydrophobic interactions. Although or metabolite must give a signal-to-noise ratio

Ann Clin Biochem 2000: 37

 696

Ann Clin Biochem 2000: 37

Paterson et al.

FIGURE 3. Comparison of ion chromatograms for morphine±TMS from a patient’s sample (top) and morphine±TMS from the in-house reference library (bottom).
 Analysis of urine for drugs of abuse 697

TABLE 1. Retention times and mass spectral data used to perform target compound analysis

Extracted ions (m/z)

Drug RT (min) Target ion Quali®er 1 Quali®er 2

Amphetamine±TFA 8´00 140 118 65

Nicotine 8´30 84 133 162
Methamphetamine±TFA 8´40 154 110 118
Ecgonine methyl ester±TMS 9´50 82 96 271
Ibuprofen±TMS 9´80 160 263 278
Paracetamol±TMS 9´90 206 80 295
MDA±TFA 9´90 135 162 275
MDA±TFA/TMS 9´95 212 135 163
MDA-d5±TFA/TMS 10´20 216 168 352
Butobarbital±TMS 10´45 341 269 300
MDMA±TFA 10´65 154 162 135
Lignocaine±TMS 10´70 86 220 235
Cotinine 10´80 98 176 118
Amylobarbital±TMS 10´85 355 300 285
Meconin 11´10 165 147 194
Pentobarbital±TMS 11´15 285 300 355
MDE±TFA 11´15 168 161 140
Carbamazepine metabolite 11´30 179 178 151
Secobarbital±TMS 11´40 297 367 339
Diphenhydramine 11´60 58 165 152
Phencyclidine 11´80 200 242 186
Caffeine 11´80 194 109 67
Orphenadrine 12´00 58 165 178
Chlorpheniramine 12´30 203 58 167
Phenobarbital 12´30 146 361 261
Carbamazepine 12´50 193 190 165
Cyclizine 12´70 99 194 165
EDDP 12´70 277 262 235
Diphenhydramine metabolite 12´90 167 183 154
Venlafaxine 13´00 58 134 121
Methadone 13´40 72 223 294
EMDP 13´45 208 115 130
Amitriptyline 13´50 58 202 215
Propoxyphene 13´60 58 91 105
Methadone±TMS 13´80 72 296 85
Cocaine 13´80 82 182 303
Mianserin 13´80 264 71 220
Trimipramine 13´80 58 249 193
Benzoylecgonine±TMS 14´00 82 240 361
Diclofenac±TMS 14´00 214 242 367
Phenytoin 14´00 281 176 25
Desmethyldiazepam±TMS 14´10 341 327 343
Oxazepam±TMS 14´50 429 313 340
Citalopram 14´70 58 238 324
Thioridazine 14´75 98 370 126
Dihydrocodeine±TMS 14´80 373 236 282
Amitriptyline metabolite 14´80 232 217 202
Dothiepin 14´80 58 202 221
Dihydromorphine±TMS 14´90 431 236 416
Desipramine±TFA 14´90 208 362 193
Procyclidine 15´00 84 98 259
Codeine±TMS 15´20 371 178 234
Diazepam 15´30 256 283 221
Lorazepam±TMS 15´30 429 431 449
Morphine±TMS 15´40 236 196 414
Chlorpromazine 15´50 58 318 272
6-Monoacetylmorphine±TMS 15´70 399 340 287
Temazepam±TMS 15´90 343 257 283

Ann Clin Biochem 2000: 37

698 Paterson et al.

TABLE 1. Continued

Extracted ions (m/z)

Drug RT (min) Target ion Quali®er 1 Quali®er 2

Olanzapine±TMS 15´90 314 301 285

Clonazepam±TMS 15´95 387 352 306
Dothiepin metabolite 16´10 204 217 235
9
-THC-11-oic acid±TMS 16´50 371 473 488
Paroxetine±TFA 16´70 138 425 109
Naltrexone±TMS (A) 17´40 542 400 242
Naltrexone±TMS (B) 18´25 485 470 388
Clozapine 18´30 243 256 192

RT=retention time; TFA=tri¯uoracetamide;TMS=trimethylsilyl; MDA=methylenedioxyamphetamine;MDE=

methylenedioxyethylamphetamine; MDMA=methylenedioxymethamphetamine; EDDP=2-ethylidine-1,5-di-
methyl-3,3-diphenylpyrrolidone (methadone metabolite); EMDP=2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline
(methadone metabolite); 9-THC-11-oic acid=11, Nor- 9-tetrahydrocannabinol-9-carboxylic acid.

TABLE 2. Limits of detection and proposed UKNEQAS clinical thresholds for the major drugs of abuse and
metabolites

Limit of detection UKNEQAS clinical threshold

Drug mmol/L mg/L mmol/L mg/L

Morphine 0´18 0´05 1´80 0´50

6-Monoacetylmorphine 0´61 0´20
Codeine 0´67 0´20 3´35 1´00
Dihydrocodeine 0´33 0´10 3´30 1´00
Amphetamine 0´74 0´10 7´40 1´00
Methamphetamine 0´67 0´10 6´70 1´00
MDA (TFA/TMS) 0´56 0´10 5´60 1´00
MDMA 0´52 0´10 5´20 1´00
Benzoylecgonine 0´35 0´10 1´05 0´30
Methadone 0´65 0´20 1´63 0´50
9
-THC-11-oic acid 0´15 0´05 0´04 0´015
Diazepam 0´70 0´20 1´75 0´50
Desmethyldiazepam 0´74 0´20 1´85 0´50
Oxazepam 1´85 0´50 1´85 0´50
Temazepam 0´66 0´20 1´85 0´50
Secobarbital 0´84 0´20 2´10 0´50

TFA=tri¯uroacetamide; TMS=trimethylsilyl; MDA=methylenedioxyamphetamine, MDMA=methylene-

dioxymethamphetamine; 9-THC-11-oic acid=11, Nor- 9-tetrahydrocannabinol-9-carboxylicacid; UKNEQAS=
UK National External Quality Assurance Scheme.

greater than 10. The limits of detection for the
major drugs of abuse and metabolites are shown
in Table 2. For comparison, the clinical cut-offs
proposed by the UK National External Quality
Assurance Scheme (UKNEQAS) for drugs of
abuse in urine are also shown. The suggested
clinical cut-off for the cannabinoid group is
0´05 mg/L, with a cut-off of 0´015 mg/L for 9-
THC-11-oic acid, the speci®c active metabolite.
As the method described here detects 9-THC-
11-oic acid, only 0´015 mg/L has been entered as
the UKNEQAS cut-off for comparison. The
method therefore has a higher limit of detection
for 9-THC-11-oic acid than that proposed by
UKNEQAS, but it has been agreed that the
method is sensitive enough for the particular
application it is designed for. The limits of
detection for all the other drugs of abuse are
acceptable for screening for drug treatment
centres.
The Toxicology Unit had previously per-
formed urine drug screening for drug treatment

Ann Clin Biochem 2000: 37

 Analysis of urine for drugs of abuse 699

TABLE 3. Recoveries of the major drugs of abuse and metabolites and concentration of drug solution used

Concentration of drug solution

Drug Recovery (%) (n=6) Relative SD (%) mmol/L mg/L

Morphine 97´3 3´5 7´0 2´00

6-Monoacetylmorphine 40´3 9´8 6´1 2´00
Codeine 91´5 1´4 6´7 2´00
Dihydrocodeine 111´2 2´7 6´6 2´00
Amphetamine 82´2 8´7 14´8 2´00
Methamphetamine 96´2 1´2 13´4 2´00
MDA (TFA/TMS) 64´6 3´2 11´2 2´00
MDMA 73´1 1´7 10´3 2´00
MDA-d5 (TFA/TMS) 82´7 8´4 10´9 0´20
Benzoylecgonine 92´3 4´2 6´9 2´00
Methadone 104´9 2´4 6´5 2´00
9
-THC-11-oic acid 36´6 5´5 5´8 0´10
Diazepam 71´1 5´0 7´0 2´00
Desmethyldiazepam 67´6 4´4 7´4 2´00
Oxazepam 31´6 4´9 7´0 2´00
Temazepam 43´6 2´6 6´6 2´00
Secobarbital 93´9 7´12 8´4 2´00

TFA=tri¯uoroacetamide; TMS=trimethylsilyl; MDA=methylenedioxyamphetamine; MDMA=methylene-

dioxymethamphetamine; 9 -THC-11-oic acid=11, Nor- 9-tetrahydrocannabinol-9-carboxylic acid; SD=stan-
dard deviation.

centres using a combination of methods. TLC approximately comparable. Using SPE±GC±MS

was used to detect opioid and other basic drugs,
and immunoassay was used for amphetamines,
cocaine metabolite, benzodiazepines, cannabi-
noids and barbiturates. Only amphetamines
were routinely con®rmed using GC with a
nitrogen-speci®c detector. This meant that a
patient’s drug pro®le was being assessed using
methods that were not legally defensible in court
should the patient ever contest the result. The
method of choice for the unequivocal identi®ca-
tion of drugs in urine is GC±MS and it is for this
reason that a method was developed using this
technique.
In today’s budgetary climate, purchasers of
laboratory services have little money to spend.
The use of SPE±GC±MS as an analytical
approach has to be seen to be as cost effective as
a more conventional approach. The more usual
method of drug screening for drug treatment
centres includes TLC for opioid and basic drugs
and immunoassay screening for amphetamines,
cocaine metabolite, cannabinoids, barbiturates
and benzodiazepines. GC with nitrogen-speci®c
detection or GC±MS is used for con®rmation of
amphetamines and for running any samples that
give unusual results by TLC. The cost of
providing the service can be broken down into
three: staff costs, consumables and equipment.
Staf®ng costs for the two approaches are
the Toxicology Unit processes up to 40 samples/
day using one technician to extract and derivatize
the samples and one mass spectroscopist to read
the chromatograms. To run a similar number of
samples using TLC/immunoassay/GC, more
technician time is needed but the input from a
chromatographer would be less. Assuming a
technician is cheaper than a chromatographer,
these two costs balance. The costs for booking-in,
reporting and checking of results are similar for
both approaches. The cost of consumables for
SPE±GC±MS is about the same as the cost of
consumables for TLC. The major expenditure
difference is that for SPE±GC±MS a dedicated
GC±MS system is needed. This compares with
TLC/immunoassay/GC, where a gas chromato-
graph is needed part time only, but there is the cost
of immunoassay reagents. If the GC±MS running
costs, including the service maintenancecost, plus
capital outlay is spread over 5 years (the minimum
time the equipment would last), the amount is less
than for immunoassay reagents alone over a
similar period.
The new GC±MS method described above is
in current use for screening urine samples from
patients attending drug treatment clinics. Using
an in-house TLC method, 10 mL of urine are
needed for a limit of detection of 1´0 mg/L for
morphine and methadone; the new method uses

Ann Clin Biochem 2000: 37

700 Paterson et al.
only 4 mL urine and is more sensitive for all the
opioid drugs. Immunoassays are only group
speci®c for benzodiazepines, opiates, ampheta-
mines and barbiturates;the new method is speci®c
for each analyte. There is no need for the two-step
approach of screen followed by con®rmation of
positives. As previously mentioned, the new
method detects not only drugs of abuse and
metabolites but also a whole range of other drugs
including antidepressants, antipsychotics and
various over-the-counter preparations. This
means it has the potential for being used to check
compliance with therapeutic medication and for
corroborating the history given by the patient.
The method also has the potentialto be used semi-
quantitatively. In addition to screening for the 61
drugs routinely analysed, other drugs speci®ed by
the clinician can be detected, provided standard
spectra are available.
In conclusion, this method shows that the
powerful technique of GC±MS can be used for
the routine analysis of urine samples from
patients attending drug treatment centres. The
information produced by this method provides
the clinician with a more complete picture of the
drug status of a patient than is currently
available by conventional means. Not only is
there increased information that is more reliable
and more speci®c than that produced using the
conventional approach of a combination of
immunoassay, TLC and GC, but the cost per
Ann Clin Biochem 2000: 37
sample of analysis using both methods remains
equivalent.
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Accepted for publication 2 March 2000