FORENSIC DETECTION

OF MARIJUANA TRACE
Thitika Kitpipit(a), Nathinee Panvisavas(a,b), Nuntavan Bunyapraphatsara(c)
a Forensic Science Graduate Programme, Faculty of Science, Mahidol University,
Rama 6 Road, Bangkok 10400, Thailand
b Department of Plant Science, Faculty of Science, Mahidol University, Thailand
c Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University, Thailand
Introduction Chemical examination is routinely used to
detect the presence of the hallucinogenic
substance in alleged materials called
tetrahydrocannabinol (THC)
Specific to Cannabis sativa
Introduction Although marijuana DNA markers have
been developed from regions such as
trnL-F [1] and the THCA synthase gene
[2], DNA analysis is not widely used in the
Thai forensic community.
Introduction In this study, we compared the use of
chemical and biological techniques to
detect marijuana materials, which had
been treated in various simulated
conditions according to the way they are
consumed.
 Materials Marijuana materials were obtained from the
Office of the Narcotics Control Board (ONCB),
and Method Thailand.
1. Leaves were boiled in water for 5 min to 8 h
2. Leaves were dried in hot-air oven, air-dried
in shade and sunlight, and burned to black
and white ashes
 Materials 3. Cannabinoids were extracted from the
treated materials and separated by TLC
and Method in hexane:dioxane:methanol (7:2:1)
petroleum ether:diethyl ether (8:2),
hexane:diethyl ether (8:2), or
hexane:dioxane (9:1)
4. DNA was extracted for PCR analysis
using the trnL-F Cannabis-specific
primer pair
Results The 197-bp DNA fragment was amplified
only in C. sativa, hence demonstrating
the specificity of the trnL-F primers
Results When testing the DNA of the treated marijuana
materials, the 197-bp trnL-F fragment was
detected in only two samples, one is the fresh
marijuana leaves boiled for 5 min and a dried-
marijuana sample
Results Comparison of four TLC solvent systems
for cannabinoid separation showed that
Rf value of the eight bands separated in
hexane:dioxane:methanol (7:2:1) were in
the range of 0.20.6, suggesting the best
separation resolution
Results TLC fingerprint of the treated marijuana
samples showed the major bands of D9-
THC, CBN, and CBD in all samples, except
the white ashes
Conclusion The study demonstrated that TLC
fingerprinting, being simple and rapid, is
a robust method for the detection of THC
in treated marijuana materials when
compared to DNA analysis.
It can detect the controlled substances in
all treated marijuana samples, except
the white ash extract.
Conclusion In contrast, DNA analysis is limited when DNA
from heat- treated materials were analyzed.
However, chemical reference standards
(controlled substances) are required for the
TLC analysis of unknown samples.
It is also suggested that DNA recovery from
the other types of samples generated should
be further optimized as there are factors
affecting the amount of DNA template
recovered from different sample types.
References A. Linacre, J. Thorpe, Detection and
identification of cannabis by DNA,
Forensic Sci. Int. 91 (1998) 7176.
M. Kojoma, H. Seki, S. Yoshida, T.
Muranaka, DNA polymorphisms in the
tetrahydrocannabinolic acid (THCA)
synthase gene in drug-type and fiber-
type Cannabis sativa L., Forensic Sci.
Int. 159 (2005) 132140.