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), which is
specific to Cannabis sativa.
Introduction
Although marijuana DNA markers have
been developed from regions such astrnL-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
and Office of the Narcotics Control Board (ONCB),
Thailand.
Method 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
and materials and separated by TLC in
hexane:dioxane:methanol (7:2:1) petroleum
Method 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

8
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 0.2
0.6, suggesting the best separation resolution

9
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

[1] A. Linacre, J. Thorpe, Detection and identification of cannabis by DNA,
Forensic Sci. Int. 91 (1998) 7176.

[2] 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.