Clinical Biochemistry 37 (2004) 175 183

Urinary screening for methylphenidate (Ritalin) abuse: a comparison

of liquid chromatographytandem mass spectrometry, gas
chromatographymass spectrometry, and immunoassay methods
J. Eichhorst, a M. Etter, a J. Lepage, a and D.C. Lehotay a,b,*
b

a
Saskatchewan Health Provincial Laboratory, Regina, SK, Canada
Department of Pathology, University of Saskatchewan, Saskatoon, SK, Canada

Received 11 July 2003; received in revised form 4 November 2003; accepted 4 November 2003

Abstract
Objective: To develop a routine method for detecting methylphenidate (Ritalin) use among drug abusers using liquid chromatography
tandem mass spectrometry (LC/MS/MS). The new methodology was designed to replace less reliable and/or more expensive and timeconsuming techniques (GC/MS and ELISA) currently employed in our laboratory, and to provide a combined one-step screening and
confirmation LC/MS/MS method.
Design and methods: Because methylphenidate abuse is very prevalent in Saskatchewan, there is a demand to provide high volume urine
screening both to detect abuse, and to monitor compliance. Random urine samples sent for drugs of abuse testing, standards, and controls
were diluted 1:100 in methanol. Diluted specimens were injected directly into an Agilent 1100 liquid chromatograph coupled to a Sciex API
2000 mass spectrometer. The method utilized selected reaction monitoring (SRM) as well as an electrospray ionization source (EIS) to detect
both urinary methylphenidate and the more prevalent metabolite, ritalinic acid (RA).
Results: There appeared to be little or no sacrifice in sensitivity because the higher dilutions exhibited much less matrix effect. Limit of
quantitation (LOQ) for methylphenidate was 100 nM and 500 nM for RA. Linear calibration curves from 100 to 1000 nM for Ritalin and 500
to 5000 nM for RA were acquired. Imprecision of spiked and true specimens did not exceed 10% and at the LOQ, it was less than 20%.
Conclusions: A rapid, sensitive, reliable, and highly specific method by LC/MS/MS for detecting methylphenidate and its metabolite,
RA, were developed. Both the cost and performance of the LC/MS/MS method were superior to GC/MS or ELISA, and it allows use of a
single rapid procedure for both screening and confirmation.
D 2003 The Canadian Society of Clinical Chemists. All rights reserved.
Keywords: Urinary screening; Methylphenidate; Ritalin

Introduction
Methylphenidate (MPH) (Ritalin) is a phenethylamine
derivative used in the treatment of depression, narcolepsy,
attention-deficit disorder, and childhood hyperkinesis [1]. It
is a central nervous stimulant that acts by inhibition of the
presynaptic uptake of dopamine and norepinephrine and by
promoting the synaptic release of dopamine [2]. Recently,
MPH has become a popular drug of abuse with the usual
mode of administration being intravenous injection of
dissolved tablets often in combination with the drug pen* Corresponding author. Saskatchewan Health Provincial Laboratory,
3211 Albert Street, Regina, SK, Canada S4S 5W6.
E-mail address: dlehotay@health.gov.sk.ca (D.C. Lehotay).

tazocine. Pentazocine (Talwin, Talacen) is a synthetic benzomorphan derivative that has properties of an analgesic
and is about one third to one sixth as potent as morphine.
The combination of Ritalin and Talwin is commonly
referred to on the street as poor mans heroin or Ts
and Rs.
The fact that MPH is commonly prescribed in the
treatment of attention-deficit disorder provides a likely
source for drug abusers. Prescribing physicians have legitimate concerns about whether or not the drug is being
administered to children or instead being sold or used
improperly by parents and guardians. In the past few years,
our Toxicology department has become burdened with
many requests for MPH analysis both for establishing abuse
and monitoring compliance. By far the largest demand for

0009-9120/$ - see front matter D 2003 The Canadian Society of Clinical Chemists. All rights reserved.
doi:10.1016/j.clinbiochem.2003.11.006

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J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

this testing is from drug treatment centres, where the

population is predominantly adult drug abusers. There
seems to be a localized preference for MPH as a drug of
abuse in Saskatchewan. Most drug treatment centres consider it to be as popular in our province as marijuana and/or
opiate abuse. The volume of requests has steadily increased
over the past 2 years to approximately 250 per week. This
requires high volume capabilities.
MPH is rapidly biotransformed in the body by hydrolysis
of the methyl ester linkage to ritalinic acid (RA), an inactive
metabolite. This hydrolysis also occurs in aqueous alkaline
solutions and during incubation with blood or plasma [1].
Because about 70% (60 81%) of MPH is eliminated in the
urine as RA, it is obviously a better indicator (more
prevalent) than the parent MPH for detecting usage. Ideally,
any assay used for urinary MPH screening should be able to
detect both parent and major metabolite. The R,RV isomer of
MPH reportedly hydrolyses more rapidly than the S,SV
isomer [3]; however, we do not attempt to distinguish
between these compounds. For some applications, it may
be necessary to differentiate between both enantiomers of
MPH as previously reported [4], but for our purposes, this
was not required. Precautions were taken to minimize
spontaneous production of RA in specimens and standards
by refrigeration and freezing if applicable.
Previous methods employed by our laboratory included
gas chromatography (GC), gas chromatography mass spectrometry (GC/MS), and an enzyme immunoassay (ELISA).
The oldest method using GC was only used sparingly and
will not be discussed here as a significant procedure.
Access to instrumentation such as liquid chromatography
coupled with tandem mass spectrometry (LC/MS/MS) has
made possible the development of high-throughput analytical methods that also provide excellent sensitivity and
specificity. Trace level determination of drugs and/or metabolites in biological matrices is an ongoing challenge in
todays clinical toxicology laboratory. Traditional techniques such as the above-mentioned GC/MS and ELISA are
very useful, but in the case of MPH detection in urine
specimens, both have drawbacks. The method developed for
GC/MS provided adequate sensitivity and specificity (using
selected ion recordingSIR); however, it required involved, labour intensive sample preparation, and with 15
20 min run times, was very slow. The ELISA method was
found to be problematic in that it produced an unacceptably
high number of false-positive results. Applying the urine
specimens directly to the 96-well plates provided a falsepositive (when compared to GC/MS) rate of approximately
10%. A modified method, which involved a simple solvent
extraction, diminished the false-positive rate to approximately 5%, which was somewhat of an improvement. The
obvious advantage over the GC/MS methodology was speed
and throughput in spite of poor selectivity.
Selected reaction monitoring (SRM) LC/MS/MS is a
reliable technique with high selectivity and sensitivity.
Analysis for analytes in bio-matrices using LC/MS/MS

has become the method of choice for many laboratories

where such instrumentation is available. A method was
developed which utilized SRM of both MPH and RA in
urine specimens with almost no sample preparation. The
urine samples, standards, and controls are simply diluted in
methanol containing an appropriate internal standard. The
dilution is done directly in a 96-well plate and mixed for
15 s before placement on an Agilent 96-well plate autosampler. This is part of an Agilent 1100 HPLC system
coupled directly to an API 2000 tandem mass spectrometer. Fifteen microliters of diluted specimen is injected
directly through the autosampler system into the electrospray source of the mass spectrometer without any chromatographic separation. Run times are 1.6 min per sample,
which allows an entire 96-well plate to be processed in
approximately 4 h. Sample preparation for 80 samples
takes only 30 min and runs can be processed overnight
if instrument scheduling is a problem.

Experimental
Methods and materials
Pure standards were obtained directly from pharmaceutical companiesMPH (Novartis), RA (Novartis), and
mepivacaine (Abbott) (Fig. 1). Levallorphan was utilized
as an internal standard for the GC/MS procedure and was
obtained from Roche.
HPLC grade methanol, methylene chloride, and isopropanol were obtained from Fisher Scientific and HPLC grade
acetonitrile (Accusolve) and ethyl acetate were obtained

Fig. 1. Structures of MPH, RA, and internal standard mepivacaine.

J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

from Anachemia Canada Inc. Ammonium hydroxide was

purchased from J.T. Baker, USA. The distilled water was
purchased locally from Arctic Glacier Inc. and the 96-well
flat-bottomed plates were purchased from Sarstedt Inc.,
USA. The plates were sealed with common aluminium foil.
All pipetting was done using adjustable Gilson pipettes
(Mandel Scientific). Certified drug-free urine was purchased
from Bio-Rad Laboratories, Canada. All patient urine specimens were acquired from random urine drug screening
samples submitted to the Saskatchewan Provincial Laboratory. Solid-phase extraction columns (World Wide Monitoring Clean Screen ZSDAU020) were purchased from
Diagnostix Inc., Canada. Both MSTFA (N-methyl-n-trimethylsilyl-trifluoroacetamide) and MBTFA [N-methyl-bis(trifluoroacetamide)] were purchased from Pierce, USA and
supplied by Chromatographic Specialties, Canada. Di-sodium hydrogen orthophosphate (dibasic) and sodium dihydrogen orthophosphate (monobasic) buffers were purchased
from BDH Chemicals, Toronto. Single-step ELISA Plate
kits for MPH (catalogue # 600515) were purchased from
Diagnostix Ltd., Mississauga, Canada.
QA material
All standards and controls were prepared using Bio-Rad
drug-free urine. Separate stock solutions (4.0 mM) of MPH
and RA were prepared in methanol and water, respectively.
Urine standards were spiked at four levels of MPH and RA
and run highest to lowest concentration both at the beginning and end of the run. Controls were prepared at or near
the proposed cut-off level for each analyte (100 nM MPH
and 1000 nM RA) as well as at a higher level more likely to
be encountered in real screening situations. These were run
every 20 samples or about 5 times on a 96-well plate.
Standards were followed by blank samples and preceded by
double blank samples.
For LC/MS/MS analysis, standard curves for MPH
included 1000, 500, 200, 100 nM standards and a blank.
Standard curves for RA included 8000, 4000, 2000, 1000,
500 nM and a blank. Mepivacaine was used as an internal
standard at a concentration of 0.03 AM in methanol. Both
positive and negative urine samples were spiked at two
levels to assess recovery. The spikes were at or near the
limit of quantitation (LOQ) for each compound and at a
higher level more realistic of urine samples from drug
abusers.
For GC/MS analysis, 4000, 2000, 1000 nM standards
and blank samples spiked in drug-free urine were run. QC
samples were spiked at 1000 nM. Confirmations of positive
samples were based on the presence of RA because of its
prevalence. A stock solution of 200 AM levallorphan was
used as an internal standard.
For the ELISA assay, a positive and negative control was
run which the manufacturer provided. We also spiked drugfree urine at 100 and 1000 nM MPH to run as controls. The
theoretical cut-off was established at 100 nM.

177

Sample preparation
LC/MS/MS
Two microliters of urine was carefully pipetted into the
bottom of the wells on a standard 96-well plate. Standards,
blanks, controls, and spiked samples were treated similarly.
To each well, 200 Al of internal standard (0.03 AM mepivacaine in methanol) was added. The plate was placed on a
Wallac Delfia plate shaker for 20 s, covered with aluminium
foil, and placed on the Agilent 1100 autosampler.
GC/MS
An extraction and modified two-step derivatization was
performed similar to previously reported methodologies [5]
(Fig. 2). Five milliliters of urine was spiked with 100 Al of
internal standard (80 AM levallorphan) and buffered with 1
ml 0.1 M phosphate buffer (pH = 7.0). Tubes were vortexed
and centrifuged at 2000 rpm for 5 min. Samples, standards,
and controls were extracted using Clean Screen drugs of
abuse cartridges. The extraction consisted of a column
conditioning step using methanol, water, and phosphate
buffer (pH 7.0). The samples were loaded, the column
washed with water, 100 mM HCl and finally with methanol.
After drying, the analytes were eluted with methylene
chloride/isopropanol/ammonium hydroxide (78:20:2). Eluent was blown to dryness with nitrogen at <40 jC. To the
dry residue, 80 Al MSTFA was added, the samples vortexed
and heated at 60 jC for 5 min. The tubes were cooled to
room temperature and 30 Al MBTFA was added. After
vortexing and heating at 60 jC for 20 min, the samples
were analyzed by GC/MS.
ELISA
To remove interfering substances that caused a significant
number of false-positive results, a simple extraction procedure was performed. To 0.5 ml samples, standards, and
controls, we added 0.1 ml NH4OH and 2 ml chlorobutane.
The tubes were vortexed vigorously for 15 s and centrifuged
at 2000 rpm for 5 min. The solvent layer was removed and
blown to dryness using nitrogen at <40 jC. To the dried

Fig. 2. Structures of MPH and RA derivatives.

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J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

extract, 0.6 ml phosphate buffer was added and the tubes were
vortexed vigorously for 5 s. The ELISA protocol was
followed from this point on [6]. One hundred microliters of
phosphate buffer was added to each well of a 96-well plate.
Twenty microliters of sample, standards, or controls were
pipetted into the appropriate wells. One hundred microliters
of enzyme conjugate solution was added to each well and the
plate incubated for 30 min at room temperature. The plate was
then washed 6 times and 150 Al of substrate added to each
well. After a 30-min incubation at room temperature, the plate
was read on a microplate reader at a wavelength of 650 nm.

5 min. Data acquisition and analysis was done using

Finnigan MassLab software v. 1.4.

Instrumentation

Results and discussion

LC/MS/MS
The MS/MS analysis was performed using an Agilent
1100 LC system including a vacuum degasser, a binary
pump, a well-plate auto-sampler, and a heated column
compartment. This was coupled to an AB MDS Sciex API
2000 triple quadrupole mass spectrometer. No chromatographic separation was performed. The mobile phase was
80% acetonitrile and 20% water containing 0.01% formic
acid. The mobile phase flow was 250 Al/min and the
injection volume was 15 Al. A 10-s flush-port needle rinse
was incorporated into the procedure to prevent cross-contamination. SRM analysis was performed using the Sciex
TurboIonSpray source at a temperature of 250 jC in positive
ion mode. The mass spectrometer parameters were optimized
using Quantitative Optimization in the Analyst 1.2 Software. The source parameters were optimized by infusing a
solution of 1.0 AM MPH, RA, and internal standard in
mobile phase at a rate of 10 Al/min using a Harvard syringe
pump. The source parameter settings were as follows: curtain
gas = 25, source temperature = 250 jC, source gas 1 = 40 psi,
source gas 2 = 40 psi, ion spray voltage = 5300 V, CAD gas =
4, CE = 30 eV. Q1 and Q3 were operated at unit mass
resolution. The SRM mass transitions monitored were
220.3 > 84.3 (RA), 234.3 > 84.3 (MPH), and 247.3 > 98.2
(mepivacaine). The dwell time for all analytes was set at 500
ms. Data were acquired and quantitation performed using
Analyst software 1.2 (AB MDS Sciex).

Fig. 3 shows an acquired set of SRM scans from a real

positive urine specimen. It is well known that reduced
electrospray sensitivity is normally caused by the presence
of endogenous components in biological matrices such as
urine. Such components will indeed cause ion suppression
especially if insufficient separation or no chromatography is
employed. Chromatography, which will separate the analytes
of interest from endogenous contaminants, is normally
required. In our method, the dilution of the urine (1/100)
was more than adequate to eliminate the ion suppression due
to contaminants, and still allow the detection of clinically
significant levels of both MPH and RA. The diluted urine
samples were sufficiently clean to be analyzed directly in the
API 2000 turboionspray source. Long runs of 100 or more
samples did not dirty the instrument; no loss of sensitivity
was observed. The simplicity and ease of sample preparation
by simple dilution allows relatively large runs to be prepared
with little analysts time. Run times of 1.6 min per sample
allowed processing of one 96-well plate in about 4 h (including autosampler delays).
The presence of either RA and/or MPH in urine indicates
the use of MPH. Because MPH is a relative newcomer to the
list of popular drugs of abuse and because its use is less
widespread, there are no specific guidelines or widely
accepted urinary cut-off levels established. We developed
our cut-off levels mostly based on assay reliability, but as
well, we took into account the levels that would be expected
after different patterns of use. Urinary MPH levels following
a single 25-mg oral dose were reported to range from 460 to
4020 nM in five adults [1]. MPH concentrations ranging
from 3.4 AM to >100 AM were found in the urine of six
arrested drivers who exhibited symptoms of hyperactivity or
drowsiness [1]. This relatively large range in expected
concentrations can at least partially be explained by the fact
that these levels have not been correlated to creatinine
concentration. It is common practice today to express random urine analytes as a ratio to creatinine concentration. Our
patient population is 99% adult drug abusers, so that a
specific correlation to dosage is not practical. With this
information in mind, we expected concentrations significantly higher than our established cut-off levels in clients who
are actively abusing the drug. As well, the fact that we can

GC/MS
The GC/MS analysis was performed using a bench top
Finnigan (Fisons) MD 800 quadrupole GC/MS (Manchester, UK). The instrument was operated in EI+, SIR mode
with dwell settings of 0.08 s and a span of F 0.10 amu. The
following m/zs were monitored: MPH180*, 181, 208,
372; RA180*, 181, 150; Levallorphan (IS)176*, 272,
355 (* Indicates quantitation ion).
Chromatographic separation was performed on a J&W
15 m, 0.25 id, 0.25 Am, DB-17 ms column (Chromatographic Specialities Inc., Canada). A GLC temperature
program was used which included an initial temperature
of 120 jC, a 1-min hold time, a 10 jC/min ramp until 200
jC, a 25 jC/min ramp until 280 jC, and a final hold time of

ELISA
The ELISA analysis was performed using 96-well plate
format kits for MPH. All pipetting was done manually with
single and multichannel pipettes and the plates were read on
a Labsystems Multiscan RC well plate spectrophotometer
(Wallac, Canada). Multicalc software was used to control
the spectrophotometer.

J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

179

Fig. 3. LC/MS/MS SRM of MPH, RA, as well as the internal standard in a urine specimen positive for MPH abuse.

detect RA which has a longer half-life (4 h compared to 2.1

h for MPH) [1], and has been reported to be 60 80 times
more prevalent in urine than MPH, gives us a reasonable
window of detection in cases of abuse (Figs. 4 6).
We did analyze a group of 262 urine samples by both
ELISA and LC/MS/MS and determined that ELISA

reported 13 specimens (5%) as false negatives. It should

be noted that 9 of the 13 false negatives contained only RA
and because the ELISA method detects only parent MPH,
the actual false-negative rate is only 1.5%.
For detection of much lower levels associated with
concerns regarding compliance in children, a simple

Fig. 4. SRM LC/MS/MS chromatogram showing data for the analysis of level 2 control.

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J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

Fig. 5. SRM LC/MS/MS chromatogram showing data for analysis of low cut-off control for RA.

solvent extraction (or SPE) can be employed which gives

us a concentrating effect providing a detection limit of
less than 0.5 nM (Fig. 7). Plasma concentrations in
children given a 10 15 mg oral dose range from 17 to
107 nM for MPH and 500 1125 nM for RA, 3 6 h after
ingestion.

Linearity
Five-point calibration curves were constructed for each
96-well plate using a linear regression based on concentration versus peak area ratio of analyte to internal
standard. A relatively small dynamic range was chosen

Fig. 6. SRM LC/MS/MS chromatogram showing data for analysis of low cut-off control of MPH.

J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

181

Fig. 7. A representative SRM chromatogram showing a 50-nM standard spiked into 0.5 ml serum and extracted using chlorobutane. The reconstitution volume
is 100 Al, which yields a concentration factor of 5. This simple extraction process eliminates most of the matrix effect and allows much lower limits of
detection.

because in urine drugs of abuse screening, there is little

or no consequence as to how high the urine levels are
above the established cut-off value. The internal standard
chosen was mepivacaine; however, it is the objective of
this laboratory to obtain isotopically labelled standards for
each species.
Ritalin: StandardsBlank, 100, 200, 500, and 1000 nM;
Linear regression (no weighting) y = 0.000246x + 0.000258
(r = 0.9994). RA: StandardsBlank, 1000, 2000, 4000, and
8000 nM; Linear regression (no weighting) y = 0.0000122x +
0.00008 (r = 0.9996).

Inter-assay precision was established over three runs

approximately 1 week apart at the respective cut-off levels
for both MPH and RA.
For the MPH cut-off of 200 nM, the following results
were obtained: %CV = 10.5, Accuracy = 98.2% (n = 27).
For the RA cut-off of 1000 nM, the following results
were obtained: %CV = 15.3%, Accuracy = 85.5% (n = 27).
Results remained consistent over the entire 4-hour sampling time of one 96-well plate.

Precision and accuracy

A total of 27 positive and negative urine specimens

obtained from drug-dependent clients were spiked with both
MPH and RA at or near the cut-off levels, and at a higher
level. All spiked specimens were assayed both before
spiking and after. The recovered amounts were compared
to theoretical values based on amount added. The results
were as follows:

The intra-assay precision was determined at three levels

for each analyte. Levels chosen were the cut-off level as
well as 50% of the cut-off, and a higher level more reflective
of actual positive urine specimens (n = 25).

Analyte

Concentration (nM)

Precision

Accuracy

MPH
MPH
MPH
RA
RA
RA

100
200
1000
500
1000
3000

CV
CV
CV
CV
CV
CV

120%
90%
103%
112%
88.5%
108%

=
=
=
=
=
=

14.9%
6.9%
2.7%
19.4%
13.3%
5.5%

Recoveries

Analyte

Concentration
spiked (nM)

Mean recovery

Precision

MPH
MPH
RA
RA

200
1000
2000
8000

106.9%
101.0%
104.8%
110.3%

CV
CV
CV
CV

<
<
<
<

20%
20%
20%
20%

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J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

Discussion and conclusion

The above-described method was developed to provide a
rapid, reliable, selective, and cost-effective method for
detecting MPH abuse among drug users. With minor modifications in sample preparation, it can also be used to
provide very sensitive detection of MPH and its major
metabolite RA in plasma to enable physicians to monitor
compliance. Both screening and confirmation are accomplished in one step using LC/MS/MS. The method is far
superior to the above-mentioned ELISA method and GC/
MS method for several reasons.

number of samples by GC/MS would be about 8 10 h. Add

to this an estimated instrument run time of 32 h (20 min/
injection) and the total time excluding data interpretation
would be more than 40 h. This definitely excludes the GC/
MS procedure as a suitable high-throughput methodology.
The ELISA method, due to the necessity of a sample clean
up step, required about 5 h to prepare a 96-well plate.
However, because the precision is such that samples had to
be run in duplicate, two 96-well plates would be required for
the same number of specimens and thus 10 h of time would
be required. If we add 20 min of plate reading time, the total
time required (excluding interpretation) to analyze 80 specimens would be greater than 10 h.

Reliability
Cost
The precision and accuracy of this method are comparable to the GC/MS method; however, it far exceeded that
of the ELISA method. ELISA specificity yielded a falsepositive rate (compared to GC/MS) of approximately 10%.
This equates to 30 false positives/week if our workload is
300 samples/week. Even with the described extraction
clean-up step, the false-positive rate (compared to GC/
MS) was only reduced to 5%. This would still provide our
laboratory with approximately 15 false positives/week.
Precision using the ELISA method was poor, so all
samples had to be run in duplicate. This added a significant cost to the analysis. Because negative results in drugs
of abuse screening do not have dire consequences, we
have little data on false negatives by ELISA. The LC/MS/
MS detection limits are similar to GC/MS SIR analysis,
but because of the absence of both extraction and derivatization steps, it provides better precision. We also encountered much better linearity when compared to GC/MS
and ELISA.
Because there is no commercially available QC material
specifically for detecting Ritalin as an abused drug, we have
produced in-house controls at two levels for both MPH
and RA. These are prepared by spiking drug-free urine with
pure standard.
Three sets of both control levels are run in each 96-well
plate. QC results are plotted on Levy-Jennings charts and
monitored as part of our laboratory QC program. The CV of
these controls was <15% when analyzed repeatedly along
with each run.
Practicality
The fact that the tandem mass spectrometry method
requires very simple and easily performed sample preparation, it is far preferable to either of the other methodologies.
A 96-well plate containing approximately 80 separate specimens can be prepared for analysis by one individual in
about 0.5 h. The instrument time is about 4 h; however; this
can be performed (after run set-up) with absolutely no
involvement by the analyst and can be left to run reliably
overnight. The sample preparation time for an equivalent

The initial expense of a tandem mass spectrometer is

very significant. Acquisition of such an instrument may
not be justified for this one assay alone; however, because
of its virtually unlimited range of applications, it can be
utilized for the analysis of a wide variety of different
compounds. Our clinical laboratory already has an operational need for this type of instrumentation in the Toxicology, Endocrinology, Neonatal screening, and Chemistry
departments. Consideration may be given to multiple
applications and shared usage to justify initial cost. Even
after these considerations, a case may be made for cost
recovery based on the following scenario. Reagents for our
previous high-throughput ELISA method cost approximately Can$3.08/test (done in duplicate). Our total staff
cost/test was approximately Can$4.00/test. Including consumables, our yearly cost (based on a volume of 200
specimens/week and not including other overhead) would
be approximately Can$80,000. This amount would almost
cover the purchase or lease of a brand new instrument
spread over 4 5 years. Because a volume of 200 samples
could be theoretically run in 1 2 days, this would leave
very appreciable time for other method development and
applications. As well, it would require about 1/8 1/6th the
staff time and labour.
Another consideration is the availability and cost of
having experienced tandem mass spectrometer operators.
Even with todays more user-friendly software, it is important to have competent, knowledgeable, formally educated staff to operate such instrumentation. The technical
skills required for clinical applications including troubleshooting and method development are definitely more
demanding than those of operating large immunoassay
analyzers. In our laboratory, we have successfully trained
two or three individuals to not only operate a tandem mass
spectrometer, but to troubleshoot problems as they develop
and to be able to develop new methods and procedures.
These individuals had an undergraduate degree in biochemistry/analytical chemistry as well as an aptitude for
computer applications and instrument maintenance and
repair.

J. Eichhorst et al. / Clinical Biochemistry 37 (2004) 175183

The method developed for determining MPH and its

metabolite RA by tandem mass spectrometry using simple
dilution sample preparation has been shown to provide a
reliable, cost-effective, and practical means of handling the
many requests we receive for detecting MPH abuse. As
well, with minor modifications, the method can also be
used for monitoring compliance.

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