FORENSIC SCIENCE

MASS SPECTROMETRY

(Principle, applications and types)

 OUTLINE

• INTRODUCTION
• APPLICATION
• BASIC PRINCIPLE
• TYPES
• SCENARIOS
• CONCLUSION
 INTRODUCTION
• Mass spectrometry is a powerful analytical technique used to quantify known
materials, to identify unknown compounds within a sample, and to elucidate the
structure and chemical properties of different molecules.

• This analytical method was developed for the quantitative analysis of some drugs
having hypnotic, sedative and tranquilizing properties like; Benzodizepine,
thioxathene, butyrophenone etc. in the whole blood, in forensic application
 APPLICATIONS
• Applications in proteomics: Characterization of proteins and protein complexes,
sequencing of peptides, and identification of posttranslational modifications.

• Applications in metabolomics: Cancer screening and diagnosis, global

metabolic fingerprinting analysis, biomarker discovery and profiling, biofuels
generation and use, lipidomics studies, and metabolic disorder profiling.

• Applications in environmental analysis:  Drinking water testing, pesticide

screening and quantitation, soil contamination assessment, carbon dioxide and
pollution monitoring, and trace elemental analysis of heavy metals leaching.
 Cont’d
• Applications in pharmaceutical analysis: Drug discovery and absorption, distribution,
metabolism, and elimination (ADME) studies, pharmacokinetic and pharmacodynamic
analyses, metabolite screening, and preclinical development.

• Applications in forensic analysis: Analysis of trace evidence (e.g., fibers in carpet,

polymers in paint), arson investigation (e.g., fire accelerant), confirmation of drug abuse,
and identification of explosive residues (bombing investigation).

• Clinical applications of mass spectrometry: Clinical drug development, Phase 0

studies, clinical tests, disease screening, drug therapy monitoring, analysis of peptides
used for diagnostic testing, and identification of infectious agents for targeted therapies.
 APPLICATIONS IN FORENSICS
It can be used in forensics for the analysis of;
• Drug or poison
• Fire debris/explosives
• Gunshot residues
Cases of murder or death due to poisoning and drug overdose are also prime target
for drug candidate analysis.
• The sample for forensics in the case of drug abuse is mainly urine, hair and blood.
• Some of the drugs in routine analysis include opiates, cocaine (C 17H21NO4),
marihuana, lysergic acid diethylamide (LSD) and amphetamines.
Types of mass spectrometry used in forensic science
• Liquid Chromatography-Mass Spectrometry: Liquid chromatography-mass
spectrometry (LC/MS) is a technique frequently used by toxicologists to analyse
substances for suspected illicit drugs and to confirm the findings of presumptive drug
tests. Analysts might test the substances themselves, or human tissue or fluid samples
to gauge whether an individual has ingested the substance, whether intentionally or not.
• Gas Chromatography-Mass Spectrometry: Many forensic labs use gas
chromatography (GC) to separate samples for MS analysis. This has been the gold
standard for many years because so many substances fragment to give a clear
fingerprint once in the gas phase. For greater resolution still, a second MS analysis can
be run (GC/MS/MS).
• Isotope Ratio Mass Spectrometry: Here, a technique such as GC is used to to
separate samples before they enter an isotope ratio mass spectrometer where specific
elements and isotopes within a sample can be analysed to determine how old a sample
is using isotopic dating.
 BASIC PRINCIPLE
• There are four stages in a mass spectrometer which we need to consider, these are
– ionisation, acceleration, deflection, and detection. Let’s go through these in
order.
• Ionisation: The sample needs to be vaporized first, before being passed into the
ionisation chamber. Here, an electrically heated metal coil gives off a stream of
electrons. The atoms or molecules in the sample are bombarded by this stream of
electrons, and in some cases, the collision will knock an electron from the
particle, resulting in a positively charged ion. Most of the ions formed have a +1
charge, as it is difficult to remove a second electron from an already positive ion.
• Acceleration:The positively charged ions are repelled from the ionisation
chamber (which is positively charged), and pass through negatively charged slits
which focus and accelerate this into a beam.
 CONT’D
• Deflection:The stream of positively charged ions are then deflected by a magnetic
field. The amount ions are deflected by depends on the mass of the ion (lighter ions
will be deflected more than heavier ones)the charge of the ion (ions with a greater
charge than +1 are deflected more)We can consider these properties as a mass/charge
ratio (m/z), where the mass of the ion is divided by its positive charge.
• Detection:By varying the strength of the magnetic field, the different ion streams
(after deflection) can be focused on the ion detector, in order of increasing
mass/charge ratio (as the lightest ions would need to be deflected the less). When an
ion hits the detector, the charge is neutralised, and this generates an electrical current.
This current is proportional to the abundance of the ion, these are sent to a computer
for analysis.
 SCENERIOS
A drug rape case involving triazolam detected in hair and urine

• University of Copenhagen, Frederik V's Vej 11, 2100, Copenhagen, Denmark.

[email protected]

In recent years, there has been heightened awareness regarding the use of drugs to modify a

person's behavior to facilitate crime. A drug rape case involving the potent, short-acting

sedative triazolam will be presented.

On three occasions, the victim consumed green tea and chocolate before being massaged and

ultimately sexually abused. Screening for alcohol, commonly used drugs and illicit

substances in blood and urine sampled during the forensic examination 20 h after the last

incident, was negative.

 CONT’D
Consequently, hair samples for chemical analysis were taken from the assaulted
individual 34 days after the last incidents.

The hair was cut into three 2-cm segments (0-6 cm) that were washed, dissolved in
extraction solvent and screened and verified by ultra performance liquid
chromatography coupled with time of flight mass spectrometry (UPLC-TOF-MS)
and with tandem mass spectrometry (UPLC-MS/MS), respectively

In the 2-cm hair segment corresponding to the period of the alleged assaults, the
presence of the sedative triazolam was revealed at a concentration of 1.0 pg/mg hair.
 CONT’D
The preserved urine sample, taken 20 h after the last incident, was reanalyzed by
UPLC-MS/MS for metabolites of triazolam, and 39 μg/l α-hydroxytriazolam was
detected in the hydrolyzed urine.

This case illustrates that hair is a valuable forensic specimen in situations where
natural processes have eliminated the drug from typical biological specimens due to
delays in the crime being reported. Furthermore, it was possible to verify the hair
finding with a urine sample by detection of a metabolite of triazolam. GC-MS
USED BY FORENSIC
 GC-MS USED BY FORENSIC EXPERTS IN MUMBAI TO
DETECT INTOXICANTS AND HELP SOLVE CRIME
 
• The Directorate of Forensic Science Laboratories (FSL) recently commented on a

case they are working on involving the death of a man in Mumbai. In this case

there is ambiguity as to whether or not the deceased man had intoxicants, such as

drugs or alcohol, in his system immediately prior to his death. Initial tests carried

out by the FSL did not indicate intoxicants in the man's system.
 CONT’D
However, the tests - carried out on a viscera sample from the man - could not be
carried out until two days after his death, due to the time taken to recover the man's
body and to send a viscera sample to the lab for testing. This delay means that the
results cannot be relied upon to be accurate as intoxicant traces are difficult to detect
in older viscera samples.

In order to accurately determine if intoxicants were in the deceased's system prior to

his death, the FSL team are using Gas Chromatography-Mass Spectrometry (GC-
MS) and High-Performance Liquid Chromatography (HPLC) which are more
successful methods for identifying traces of intoxicants in older viscera samples.
 CONT’D
Once forensic tests have been carried out using GC-MS and HPLC, the FSL will be
able to confidently conclude if intoxicants were present in the deceased's system
when he died.

Now that the FSL are using Chromatographic techniques, such as GC-MS and
HPLC, to test viscera samples, it will be possible to more accurately determine
causes of death, as information which would have previously been overlooked, such
as the presence of intoxicants in older viscera samples, will now be discovered.
 Aarey colony murder case: Mumbai forensic experts using
‘high-end instruments’ to detect intoxicant traces
• Mumbai: The forensic scientists are using all possible methods to ascertain if EOW
officer’s lone son Atharva Shinde was intoxicated during the rave party, as no
traces of drugs and alcohol were found in his ‘viscera sample’ during the normal
method of Chemical Analysis (CA) test.
• A senior FSL official told the Free Press Journal that Atharva’s viscera
sample was sent a bit late. “He (Atharva) died on May 8 and his body was
recovered on May 9 around 9:30 am. His viscera sample was collected
same day in the evening and was sent to our lab on May 10. The traces (of
alcohol or drugs) are easily not found if the viscera sample is sent late. In
this case too, we did not find any trace of alcohol or drugs during normal
CA test of the sample. So before submitting negative report on alcohol or
drug, we have to be 100 percent sure from our end. Hence, we are now
using high-end instruments to detect the traces of intoxicants,” said the
senior FSL officer.
 Cont’d
The forensic scientists are using their powerful gold standard of technique called Gas
Chromatography and Mass Spectrometry (GCMS) to analyse if Atharva died of lethal
cocktail of liquor and drugs which were allegedly served in the rave party at Royal
Palms’ bungalow number 212 on May 7 night.
“We are also using High-Performance Liquid Chromatography (HPLC) instrument to
separate, identify and quantify each component found in the (viscera) sample. If the
report comes negative, we will use advanced technique called High-Performance
Thin-layer Liquid Chromatography (HPTLC) to find the traces of alcohol and drug, if
any,” said another FSL officer. At last, the FSL will use its stas-otto method to analyse
if pico-gram level of intoxicant is detected. “All these tests will take at least four to
five more days,” the FSL officer said. A separate team of forensic scientists are also
analysing Atharva’s clothes to find the traces of alcohol and drug. The FSL team is
also checking if ‘foreign biological fluid’ is found on his clothes as his father has
alleged that Atharva was sexually assaulted as there were lacerations on his private
part.
 CONCLUSION
• In forensic study, sample is in minute quantity; therefore,
high sensitivity is required for analysis. Mass spectroscopy
coupled with gas chromatography emerged as an
indispensable tool in forensic field as well as LC–MS has also
wide utility in forensic study.
• In forensic studies, the use of mass spectroscopy is becoming
significant because of increase in the demand to investigate
use of illegal drugs through analysing body fluids and tissues.
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Bioanal. Chem. 394:1985–86 5.

• MnookinJL,ColeSA,DrorIE,FisherBAJ,HouckMM,etal.2011.Theneedforaresearchcultu
rein the forensic sciences. UCLA Law Rev. 58:725–79 6.

• Bono J. 2011. Commentary on the need for a research culture in the forensic sciences.
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