Alex Olzinski

SCH 354 02 Instrumental Analysis

17 November 2017
Analysis of Drugs in Hair Samples Using GC/MS

In today’s society, many people believe that there is a drug problem. Common street
drugs, like heroine, cocaine, methamphetamines, and others, are becoming more readily
available and easier to purchase. Many companies are requiring future employees to pass a drug
test in order to be hired, and addicts in rehabilitation programs are often tested to ensure they are
not using. With this, the reliability of urine tests is not always accurate, as the drugs can only last
for a few hours or days.2 This is why drug analysis in hair has become more popular in forensic
and clinical toxicology.1
One of the best methods for analyzing drugs in hair samples is gas chromatography
coupled with mass spectrometry (GC/MS). GC/MS combines two analytical techniques that are
useful for the quantification of a range of samples. GC separates molecules in a sample by their
retention time in a column, and MS identifies the individual molecules.
In gas chromatography, a liquid sample is injected into a column. Columns are set up to
either have a polar stationary phase (SP) or a non-polar stationary phase. Non-polar columns
typically contain polysiloxanes as the SP, while polar columns contain polyethylene glycol as the
SP.3 The interactions between the sample and the SP influence the retention time of the sample in
the column; polar molecules will stay in a polar column longer than non-polar molecules, and
non-polar molecules will stay longer in a non-polar column than polar molecules. The sample
injector and column are both heated to high temperatures by the oven to volatize the samples and
allow them to flow through the column.3 The sample is also carried through the column with the
aid of an inert gas.3

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Figure 1: Block Diagram of a Gas Chromatograph 6
The separated molecules then individually enter into the mass spectrometer. These
instruments are comprised of an inlet, ionization source, mass analyzer, ion detector, and a
readout device.3 In GC/MS, the inlet is replaced by the outlet of the column in GC.3 The
ionization source bombards the molecules with electrons, causing them to form ions.3 Different
ionization sources are used depending on what samples are being tested. Gas phase ionization
sources are good for thermally stable, and volatile, samples; examples are electron ionization
(EI) source and chemical ionization (CI) source.3 Desorption ionization sources are good for
non-volatile, thermally unstable compounds; examples are matrix-assisted laser desorption
ionization (MALDI) source and electrospray ionization (ESI) source.
When molecules ionize, they are not always stable. Unstable molecules reduce their
energy by fragmenting, or breaking off into pieces.3 These charged pieces all have varying
masses, which the mass analyzer will filter them by mass.
The second component of MS is the mass analyzer. This part passes the ions from the
ionization source through an electromagnetic field, filtering them by mass.3 The magnetic field
can be changed to allow a certain mass to pass through, which is how the detector can read the
individual ions. Examples of mass analyzers include quadrupole mass analyzer, ion trap mass
analyzer, and time-of-flight (ToF) mass analyzer.3
The third component is the ion detector. These are typically discrete-dynode electron
multipliers or continuous-dynode electron multipliers.3 These ion transducers take an ion and
convert it into an electric current that is amplified and converted into raw data.3 The raw data is
sent to a readout device and translated into a mass spectrum, and the spectrum can be analyzed to
determine what compound is in the sample.

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Figure 2: General Block Diagram of a Mass Spectrometer
Because this method can determine compounds in a matrix, GC/MS is employed in
detecting trace amounts of drugs in hair. When a person uses a drug, their body processes that
drug just like any other chemical that person puts into their system. The processed drugs go
through the blood stream, and some of it is deposited into the hair as hair strands grow.1 Trace
amounts of drugs can also enter into hair strands by the hair coming into contact with the drugs
directly, or even through inhaling residual drugs.1
Analyzing hair samples for drugs also poses more benefits. Using hair is less invasive to
a person, the sample can be stored at room temperature with no other specific conditions, and
there is less likely of a chance of contamination of the sample.1 Also, because hair grows at about
1-2 cm a month, by dividing a hair sample into 1-2 cm lengths, a single sample can tell how long
a person has been using drugs.1
Not only can hair analysis be used to tell how long a person has been using drugs, but
also what drugs they have been taking. This approach often looks for methamphetamines,
cocaine, ketamine, methadone, and ∆9-tetrahydrocannabinol (THC), among others.4 Techniques
have also been developed for quantifying cannabidiol (CBD), cannabinol (CBN), and sometimes
just detecting 11-nor-∆9-tetrahydrocannabinol-9-cacrboxylic acid (TCH-COOH).5
One of the problems with hair analysis is the extraction of the drugs from the sample.
One method that is typically used is by digesting the hair in an acid or base. The hair is placed in
either sodium hydroxide or hydrochloric acid, and the solution needs to be heated to extract the
drugs; however, the high temperature can cause the adverse effect of degrading the drugs.1 If the

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drugs are degraded, then the exact quantification cannot be determined since the altered drugs
will not appear the same in a mass spectrum as the drugs.
A technique was developed to reduce the degradation of drugs in either acid or base. A
study looked at digesting the hair samples using an enzyme rather than an acid or a base.1 The
study placed hair samples containing THC and cocaine in a solution with Proteinase K enzyme,
which allowed the hair samples to be digested at a lower temperature, thus not degrading the
drugs.1 Quantification was determined by spiking the sample with THC-D3 and cocaine-D3 as an
internal standard, and the ratio of internal standard peak height to analyte peak height was
calculated.1 The study was simultaneously conducted using sodium hydroxide, and the results
showed that enzymatic digestion had a lower limit of detection (LOD) and limit of quantification
(LOQ) for THC and cocaine.1
Another study looked at completely automating the analysis of hair samples, from
preparation to instrumentation. The study used a MultiPurpose Sampler (MSP) with
interchangeable modules to perform the entire analysis by instrumentation alone.2 The MSP was
equipped with a shaker for sample extraction, a centrifuge for phase separation, an evaporation
station under controlled vacuum and temperature, and a solvent filling station.2 The automation
technique was compared against a manual technique to validate the study.2 The study found that
the automated method had lower LOQ’s compared to the manual and to similar literature, and
that the automated method could run more samples in a shorter amount of time compared to the
manual method.2
This last study looked at attaching a MPS to a GC/MS to fully automate the analysis.
Another study combined headspace solid-phase microextraction (HS-SPME) with GC/MS.4 The
previous studies all focused on quantifying THC, CBD, and CBN. This study looked at
simultaneously determining and quantifying amphetamines, ketamine, methadone, cocaine,
cocaethylene, and THC.4 The method had LOD’s ranging from 0.01 to 0.012 ng/mg, and LOQ’s
of 0.02 to 0.37 ng/mg.4 The method was even applied to real hair samples taken from volunteers,
and most drugs found in the samples were able to be quantified, while some were just detected.4
Another study combined enzyme-linked immunoassay (ELISA) with GC/MS.5 ELISA
was used as a screening for drugs, while GC/MS was used as a confirmation; this combination is
useful for forensic applications.5 This method can also simultaneously detect multiple drugs from
a single sample, including THC, CBN, and CBD.5 The advantage of this method is that only one

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sample preparation is needed to be performed, since both ELISA and GC/MS can use the same
prepared sample for analysis.5
These four studies have shown different sample preparations and/or instrumentation
additions, but all employed GC/MS to determine and quantify illicit drugs in hair samples. The
research has found that automating the analysis is faster, runs more samples per time, and greatly
reduces error; simply put, the hair samples only need to be manually washed, ground, and placed
in a vial with an internal standard, and the instrument does the rest.2 By automating the process,
random error and gross/personal error are reduced, and if there is any systematic error, that can
be corrected.
The research also found that dissolving hair samples in an enzyme reduced drug
derivatisation compared to the traditional methods of dissolving in acid or base at high
temperatures.1 This is especially important when analyzing THC, since the acid or base reacts
with THC to form CBN, CBD, and THC-COOH. The problem is that THC-COOH can only be
quantified with tandem mass spectrometry, which many facilities are not able to perform.2 Unless
an enzyme is used, the quantification of THC has to be determined by looking at THC, CBN, and
CBD, and even then it is not fully accurate because of the loss from THC-COOH.
Lastly, the research found that coupling GC/MS with HS-SPME improved overall
accuracy, while needing only a small amount of hair sample.4 The HS-SPME uses a fiber that
adsorbs volatized drugs in the headspace of the vial, that way the sample can be used again for
future testing.4 This is useful for clinical and forensic testing, since retesting samples is always
possible.
Hair analysis by use of gas chromatography/mass spectrometry for the detection and
quantification of illicit drugs is a very important method for a wide range of applications. In
forensics, this can prove that a suspect has been in contact with a certain drug, which can place
them at a scene. It can also show what drugs a victim has been using, which can differentiate
between a suicide by overdose or murder. In clinical cases, a recovering addict can prove they
have not used a drug since rehabilitating, because hair analysis can cover a longer time period
than blood or urine tests.
Although the actual GC/MS aspect does not change from analysis to analysis, the
techniques employed for preparation can vary. This research has shown that the best way to
determine and quantify illicit drugs in hair sample is to have the whole method automated to

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reduce error, to digest the hair samples with enzymes to reduce degradation of the drugs, to
couple the method with techniques like HS-SPME to reduce sample consumption, and, specific
to forensics, also have a screening technique, like ELISA, to confirm findings.
Future scientific research could be done to see if this process is feasible. While reducing
the error from this process, the time, efficiency, and cost should also be factored in as well.
GC/MS is a widely used analytical technique because of its versatility, but falls short when
samples are not volatile. Therefore, another method should be developed using high-performance
liquid chromatography (HPLC) using these ideal conditions to ensure an even greater application
of hair analysis.

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 Works Cited
1. Breidi, S. E.; Barker, J.; Petróczi, A.; Naughton, D. P. Enzymatic Digestion and Selective
Quantification of Underivatised Delta-9-Tetrahydrocannabinol and Cocaine in Human
Hair Using Gas Chromatography-Mass Spectrometry. J. Anal. Methods Chem. 2012,
2012, 1-7.
2. Heinl, S.; Lerch, O.; Erdmann, F. Automated GC-MS Determination of Δ9-
Tetrahydrocannabinol, Cannabinol and Cannabidiol in Hair. J. Anal. Toxicol. 2016, 40,
498-503.
3. Hoover, D. A. (2017). Lecture. CH 354 Seton Hill University . Greensburg.
4. Merola, G.; Gentili, S.; Tagliaro, F.; Macchia, T. Determination of different recreational
drugs in hair by HS-SPME and GC/MS. Anal. Bioanal. Chem. 2010, 397, 2987-2995.
5. Tassoni, G., Cippitelli, M., Ottaviani, G., Froldi, R., Cingolani, M. Detection of
Cannabinoids by ELISA and GC-MS Methods in a Hair Sample Previously Used to
Detect Other Drugs of Abuse. J. Anal. Toxicol. 2016, 40, 408-413.
6. https://teaching.shu.ac.uk/hwb/chemistry/tutorials/chrom/gaschrm.htm (accessed Nov 14,
2017).