Forensic Chemistry Bell Second Edition

Forensic Chemistry

Suzanne Bell
Second Edition
Pe a r son Edu ca t ion Lim it e d
Edinburgh Gat e
Harlow
Essex CM20 2JE
England and Associat ed Com panies t hroughout t he world

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ISBN 10: 1-292-02044-X

ISBN 13: 978-1-292-02044-0

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 Instrumentation

Stretched
O
C

Nominal
X X X
separation

O O O O

C C C Compressed

Vibrational frequency
2000 cm–1

+
Excitation frequency
9395 cm–1 FIGURE 35 Frequency of a hypothetical chemical
bond vibration. This bond expands and contracts
at a frequency of 2000 cm–1. How polarizable the
bond is will determine how it scatters incoming
light. At the maximum stretch, the polarizability of
the bond will not be the same as when the bond is
at maximum compression, so polarizability has the
Modulation same frequency as the vibration.

and hydrogen are relatively small, and the bonds in water are not easily deformed by
interaction with visible light. Because visible light is used for excitation, samples can be
mounted on or contained in glass.
Inelastic scattering can result from interactions with the polarizable bond
that produce signals (Stokes and anti-Stokes) at the excitation wavelength plus or
minus the vibrational mode of the bond, as shown in Figure 36. The Stokes signal
is the stronger of the two, but it is still several orders of magnitude weaker than the
Rayleigh line. Further problems can arise when higher energy excitation sources,
such as those in the visible and UV range, are used. These sources can cause fluores-
cence (Figure 37), which may produce a signal that will swamp the weak inelastic
scattering signal. Despite these issues, Raman spectroscopy is finding a wider niche
in forensic applications, given that it provides information that is complementary to
absorption IR and is generally nondestructive.

3 MASS SPECTROMETRY
3.1 Overview
The name mass spectrometer/mass spectrometry (MS) is somewhat of a descriptive
misnomer. Mass spectrometry works on the basis of separation, but not separation of
electromagnetic energy into component wavelengths. Rather, a mass spectrometer dis-
perses mass fragments so in this sense, “spectrometer” is a reasonable analogy, since
the output is a spectrum of masses across a range of masses; an MS “scans” masses in
the same sense that spectrometers scan electromagnetic energy. A good way to visualize

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 Instrumentation

Inelastic Inelastic

Elastic
scatter

Intensity
cm–1
11395 9395 cm–1 7395

E2 Second excited state

1064 nm

E1 First excited state

E0 Ground vibrational state

Anti-stokes Rayleigh Stokes

∆E (E2 – E0) = 1064 nm = 9395 cm–1

∆E (E1 – E0) = Shift = 2000 cm–1
FIGURE 36 A Raman spectrum of
a sample bond in a system using a
near-infrared laser as the excitation
source. Elastic scattering or Rayleigh 2000cm–1
scattering dominates the spectrum,
with inelastic scattering at wave
numbers plus or minus the equivalent
of the vibrational frequency.
The Raman spectrum can also be
normalized and plotted as the shift in 0 1000
wavelength of the signal. Shift (cm–1)

a mass spectrometer is as a mass filtering device, as shown in Figure 38. When this
device is coupled to the output of some type of sample introduction system such as a
gas chromatograph (discussed below), the flow is directed into a vacuum region, where
the sample is ionized and fragmented to variable degrees. The ions are then introduced
into a filtering device that separates them on the basis of their mass-to-charge ratio.
Masses are often specified in units of Daltons (Da) and the ratio of mass to charge is
represented by m/z. Ions arrive at the transducer and are converted to electrons. The
signal is amplified by an electron multiplier and recorded.
E2

FIGURE 37 The fluorescence problem. If the

laser is at a wavelength with sufficient energy hυ
to promote the molecule to the next higher E1 Fluorescence
state, the molecule can decay to a metastable
state and then fluoresce, producing a signal
that will swamp the weak scattering signal. E0

180
 Instrumentation

+ –

+ + + +
+ +
+++ + + + + + +
+ + ++ + +
+ ++ + + +
+ + +
+ FIGURE 38 A generic mass
+
spectrometer, which does with mass
fragments what a grating does to
Source Mass analyzer Transducer/detector light: filters it and separates it into
(electron multiplier) individual components.

3.2 Basic Designs

There are many types of ionization modes and mass separation filters, the discussion
of which is beyond the scope of this book. In forensic science applications, one of the
most common MS designs is the electron impact ionization/quadrupole mass filter
(Figure 39). Ionization and fragmentation are achieved by the collisions between mol-
ecules from the sample and electrons generated by a filament, as shown in Figure 40.
This ionization technique is referred to as electron impact (EI), and it is considered to be
a hard ionization technique. This means the fragmentation is fairly complete as opposed
to a soft ionization technique. We will discuss an example of soft ionization shortly. In
EI mode, few collisions result in ionization, but enough to generate both positive and
negative ions, with positive ions usually being the polarity of interest. The positive ions
are pushed into the focusing lenses by a repeller plate kept at a positive potential. The
degree of fragmentation depends on the electron energy; standard values are 70 eV. The
vacuum is necessary to prevent secondary collisions and combinations. Ions are focused
into a tight stream by a series of electronic lenses and introduced into the quadrupoles,
where alternating DC and radio-frequency currents determine the field and thus the ion
pathways. At a given setting, only ions with a particular mass transit the quadrupoles
safely, whereas all others collide with the quadrupoles or are ejected.

Representative Resonant ion

nonresonant passed through ilter
ion paths

Transducer

Accelerating
electrodes +
Ion source
– –
FIGURE 39 A quadrupole mass
+ spectrometer. Alternating DC and
radio frequencies applied to the
quadrupoles dictate ion paths. At
any given setting, only one mass
Sample will get through and this is called
the resonant mass.

181
 Instrumentation

e– target (+) Electronic lenses;

Repeller (+) lens stack

Neutral
e–
molecules
e–
Sample + +
Ions
entrance + + + +
+

FIGURE 40 The inlet, ionization region, and

lens stack of a typical mass spectrometer. The e– stream
mechanism of ionization shown here is electron
impact (EI). Filament

Under standard EI conditions (70 eV), compounds are identified through the mass
fragmentation pattern, library matches, and traceable standards. Key to identification
is reproducible and controlled instrumental conditions. Electronics must be set such
that patterns are reproducible in and across instruments. This is done by an internal
calibration or tuning procedure in which a standard compound, usually perfluorotribu-
tylamine (PFTBA), is introduced into the mass spectrometer and settings are adjusted
or tuned until the resulting mass spectrum meets the required criteria of masses
detected and relative abundances. The standard forensic library is the NIST/EPA/NIH
Mass Spectral Library, which was acquired under instrumental conditions for PFTBA.
Because the electronic settings alter mass intensities, if an instrument is out of tune (i.e.,
if it cannot produce a PFTBA spectrum with the required mass peaks and intensities),
any calibration curves obtained under those tuned conditions have to be redone.

3.3 ICP-MS
For elemental analysis, particularly of the metals and semimetals, a mass spectrometer
has obvious appeal; the challenge is in ionizing the sample prior to introducing it into
the quadrupole (or other mass filter). The solution was the development of inductively
coupled plasma (ICP) torches, which were described in the 1970s and became avail-
able as ionization sources for mass spectrometers in the 1980s. A schematic of an ICP
torch is shown in Figure 41. Within the torch, there are three concentric quartz tubes.
Argon flows through the tubes as shown, consuming several liters a minute of the gas.
At the top of the torch are water-cooled induction coils that are powered by a radio fre-
quency generator. The torch is ignited by a spark from a Tesla coil, which generates ions
that then flow through the rapidly oscillating magnetic field generated from the coil.
Frictional and collisional interactions heat the plasma (consisting of Ar+ and electrons)
to temperatures in excess of 6000 K. Sample that flows with argon through the center of
the coil experiences temperatures of ,6500 K, resulting in rapid de-solvation, dissocia-
tion, atomization, and ionization.
The plasma stream cannot be directed into the mass spectrometer owing to the
high temperature and flow rates. Thus, the design of the interface region is a primary
technical challenge in ICP-MS. As shown in Figure 42, a differentially pumped design is
used. The plasma is directed onto an orifice into a region that is maintained at a vacuum
of about 1 torr. The sample cone allows a small stream of the plasma to enter, where
it then passes through a second orifice called the skimmer cone and on into the high
vacuum region. The beam is focused by a series of electronic lenses, some of which may
direct the ion beam off-axis. This reduces the number of neutral species in the beam.

182
 Instrumentation

Approximate
Approximate height
temperature above coil
1800 K 25 mm

2000 K 15 mm

Hottest part
of plume
3000 K 0 mm

Coil of metal tubing

(water coolant inside)
for radio-frequency
power input

Top view of plume

at coil top

Silica
glass tube

Plume
Metal
tube coil

FIGURE 41 A plasma torch for ICP. A radio-

Coolant Gas (Ar) frequency generator ionizes argon in the tube
Ar gas for plasma and accelerates the ions, maintaining heat by
Sample sustained collisions. Once the generator is off,
aerosol the plasma stops forming.

Skimmer cone
Sample
cone ~10–5 torr

Laser Torch
ablation
Ion
optics Collision
cell Quadrupole Detector

Nebulizer

Liquid Gas supply (He, H2)

~1 torr

Digestior

FIGURE 42 Schematic of a generic ICP-MS instrument. Sample can be introduced into the torch via
pumping of liquid samples or by laser ablation. Note the collision cell is off-axis.

183