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ISSN: 2692-5206, Impact Factor: 12,23
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FORENSIC DETECTION OF CANNABINOIDS IN BIOLOGICAL FLUIDS:
TECHNIQUES AND SENSITIVITY
BARAKAYEV LAZIZBEK DJALOLOVICH
Expert of the forensic chemistry department of the Bukhara branch of the Republican
Forensic Medical Expertise Scientific and Practical Center
JUMAYEVA SHARIFA JURAYEVNA
Expert of the forensic chemistry department of the Bukhara branch of the Republican
Forensic Medical Expertise Scientific and Practical Center
QAYUMOV BAXTIYOR ALLAMURODOVICH
Expert of the forensic chemistry department of the Surkhandarya branch of the
Republican Center for Forensic Medical Expertise
Abstract
Forensic investigations often rely on the detection of cannabinoids in biological fluids to establish
drug use and intoxication. While existing literature covers individual methods for cannabinoid
analysis, a comprehensive guide detailing procedures for various sample types and their respective
sensitivities remains absent. This study addresses this gap by presenting detailed protocols for
isolating cannabinoids from saliva, blood, urine, and skin swabs. Extraction techniques with
various solvents are employed, followed by thin-layer chromatography (TLC) for detection. The
study demonstrates the effectiveness of urine as a readily accessible and informative sample for
cannabinoid analysis. The TLC method boasts a sensitivity of 0.1 micrograms, providing a reliable
means for identifying cannabinoids with high precision. These procedures and sensitivity data
contribute significantly to advancing forensic science, offering valuable resources for practitioners
and researchers in the field. The study emphasizes the importance of urine as a highly informative
and readily accessible sample for cannabinoid analysis in forensic investigations. This
comprehensive guide provides a valuable resource for forensic practitioners and researchers,
contributing to the advancement of forensic science.
Keywords
: Cannabinoid detection, Forensic science, Forensic toxicology, Drug analysis,
Biological fluids, Urine analysis, Blood analysis, Saliva analysis, Skin swab analysis, Thin-layer
chromatography (TLC), Tetrahydrocannabinol (THC), Cannabinol (CBN), Cannabidiol (CBD)9
INTRODUCTION
The detection of cannabinoids in biological fluids is crucial for forensic investigations, particularly
in cases involving drug use, intoxication, and related criminal activity. While various analytical
techniques for detecting cannabinoids exist, a comprehensive guide outlining specific procedures
for different sample types and their respective sensitivities remains absent. This gap in knowledge
hinders the efficiency and accuracy of forensic investigations relying on cannabinoid detection.
This study addresses this gap by presenting a practical guide to forensic chemical detection of
cannabinoids in various biological fluids. We detail procedures for isolating cannabinoids from
saliva, blood, urine, and skin swabs using extraction techniques with different solvents, followed
by thin-layer chromatography (TLC) for detection. Our findings demonstrate the effectiveness of
urine as a readily accessible and informative sample for cannabinoid analysis, emphasizing the
sensitivity and reliability of the TLC method for achieving precise and conclusive results. This
study provides valuable insights into the practical aspects of forensic cannabinoid detection,
offering a valuable resource for forensic practitioners and researchers to enhance their
investigative capabilities.
INTERNATIONAL JOURNAL OF ARTIFICIAL INTELLIGENCE
ISSN: 2692-5206, Impact Factor: 12,23
American Academic publishers, volume 05, issue 02, 2025
https://www.academicpublishers.org/journals/index.php/ijai
page 522
The Republican Scientific and Practical Center for Forensic Medical Examination of the
Surkhandarya branch is located in the Surkhandarya region in the city of Termez.
Surkhandarya region is the southernmost region of Uzbekistan
The administrative center is the city of Termez.
The central and southern parts of the region are flat. The Gissar range is located in the north, in
the west and northwest are its spurs - the Baisuntau Mountains (4425 m) and Kugitangtau (3139
m), in the east is the Babatag range (up to 2290 m), in the south is the valley of the Amu Darya
River. In the south it borders with the Balkh province of Afghanistan, in the southwest - with the
Lebap Velayat of Turkmenistan, in the northwest - with the Kashkadarya region, in the east - with
the Nosiri-Khusrav and Shaartuz districts of the Khatlon region of Tajikistan, in the northeast -
Tursunzadev, Rudaki and Shakhrinavsky districts of Tajikistan, in the north - with Ainin and
Penjikent districts of the Sughd region of Tajikistan. As of August 1, 2022, the population of
the Surkhandarya region numbered 2,771,100 people - this is the 7th place (out of 14) among the
regions of Uzbekistan. The region consists of 1 city of regional subordination (Termez) and 14
districts (Angora, Bandikhan, Baysun, Denau, Dzharkurgan, Kizirik, Kumkurgan, Muzrabad,
Altynsay, Sariasi, Sherabad, Shurchinsky, Termez, Uzun)
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In our branch of the Scientific and Practical Center for Forensic Medical Examination of the
chemical department, various analyses are carried out, in most cases, this is a study on
cannabinoids (hashish, anasha, etc.) Cannabinoids are a group of chemical compounds that have a
psychoactive effect on the human div. Most of them are of natural origin and are found in
cannabis. Synthetic-synthesized cannabinoids are less common.
The main sources of cannabinoids are marijuana, hashish, and anasha (ganja). They differ from
each other in the concentration of the active substance and the method of preparation.
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Cannabinoids are drugs and are of plant origin. The main psychoactive effects of hashish and
marijuana are associated with 9-delta-tetrahydrocannabinol (THC), a narcotic substance contained
in the resinous substance, stems and leaves of female specimens of wild cannabis (Cannabissativa).
A variety of wild-growing cannabis, Indian hemp (Cannabisindica), is especially rich in THC.
Forms of cannabis used in illegal sale: marijuana; hashish; hashish oil. Marijuana is the dried and
crushed upper part of the cannabis plant with leaves and flowers. The content of psychoactive
substances in marijuana reaches 13-15%. Hashish resin (tar) released by cannabis during the
growing season has a green, dark brown or black color. The content of the main psychoactive
substance (THC) is usually about 2%, but can reach 9-10%. Hashish oil is a concentrated dark
liquid and viscous cannabis resin extract with a THC content of 10 to 30-60%. Hashish oil is added
to regular tobacco cigarettes or cigarettes with marijuana (in 1 cigarette 500-750 mg of marijuana
with a THC content of 1-4%).
The effect of cannabinoids on the human div
The effect of cannabinoids is to stimulate the active release of a neurotransmitter in the brain called
dopamine, which causes a feeling of euphoria. Although the effect of a psychoactive substance on
the div is individual and can vary from one person to another, most people who use cannabinoids
experience the following sensations:
euphoria;
mental and physical relaxation;
altered perception of time;
increased sensory perception;
pain reduction;
increased appetite.
The side effects of cannabis include:
the occurrence of coughing attacks; anxiety;
chest and lung discomfort; the feeling of "buzzing" of the div;
redness and dryness of the eyes; rapid heartbeat;
dizziness, fainting; increased sweating;
impaired concentration;
hallucinations, paranoia and psychosis (in case of overdose).
Forensic definition of cannabinoid intoxication
When examining living persons for the presence of cannabinoids, the main object of research is
urine. Urine is at the same time the most accessible and convenient for chemical and toxicological
examination.
The main component determining the narcotic psychoactive properties of marijuana is trans-delta-
(9)-tetrahydrocannabinol (THC). The components that confirm the use of marijuana are THC and
its
main
metabolites:
the
primary
psychoactive
metabolite
11-hydroxy-delta-(9)-
tetrahydrocannabinol (11-OH-THC) and the final biologically inactive product 11-nor-9-carboxy-
delta-(9)-tetrahydrocannabinol (THC-COON), as well as in smaller amounts of cannabinol and
cannabidiol .
The primary metabolite can be detected in blood plasma; in urine, it can be detected only in the
first few hours after ingestion and in much smaller amounts.
To determine cannabinoids in urine, we initially performed an analysis using
immunochromatographic urine analysis, which is a test for the detection of a number of
psychoactive compounds.
The choice of a liquid medium such as urine for analysis has certain advantages. The result can be
obtained quickly and with a high degree of informativeness and reliability due to the rather high
INTERNATIONAL JOURNAL OF ARTIFICIAL INTELLIGENCE
ISSN: 2692-5206, Impact Factor: 12,23
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page 525
concentration of the studied substances in the urine. Having enough urine for analysis is also not
a problem. In addition, this method has been worked out and is the leading one in the primary and
emergency examination of a person for drug use, including cannabinoids. The method of analysis
is immunochromatography, which allows detecting 14 types of narcotic substances in urine with
an accuracy of 90%.
Forensic chemical detection of cannabinoids in biological fluids. Description of the procedure:
• Technique for isolating cannabinoids from saliva
A. The resulting sample is mixed with 50 ml of saturated aqueous sodium chloride solution B.
cannabinoids are extracted with ethyl acetate (or ethanol) three times 10 ml for 5 minutes. The
extract is dried by adding 1-1.5 g of anhydrous sodium sulfate for 30 minutes. The extract is filtered
through a paper filter and evaporated into several drops
• The technique of isolating cannabinoids from the blood
A. 5 ml of blood plasma is extracted four times with a mixture consisting of petroleum ether and
1.5% pentanol by volume (5 ml each) B. The combined organic layer is evaporated to a volume of
several drops and transferred to a plate. B. Chromatographed under the same conditions as the
extract from saliva.
• The technique of isolating cannabinoids from urine.
A. 1 ml of methanol and 150 ml of 50% caustic soda solution are added to 2 ml of urine and mixed.
B. The bottle is tightly sealed and placed in a thermoblock for 10 minutes at 60 degrees B. After
cooling, the vial is opened and 6 n. hydrochloric acid is added to pH = 2-3 g. Twice extracted with
a mixture of B (n-hexane and ethyl acetate (7:1) mixed in a ratio of 7:1) 5 ml each. The top layer
is separated, transferred to a vial, evaporated to a dry residue in a stream of warm air
• The technique of isolating cannabinoids from flushing
A. Sampling (flushing) from the surface of the fingers and lips is carried out separately, by wiping
with a cotton or gauze swab moistened with ethyl alcohol B. Alcohol from the samples is
evaporated at room temperature. B. Cannabinoids are extracted from the tampon material with
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ethyl acetate (ethanol) twice in 10 ml for 1 min . The combined extract is evaporated to a final
volume of 0.1-0.3 ml
• Detection of cannabinoids by thin-layer chromatography
A. The dry residue obtained after evaporation of the final volume of the extract is dissolved in
chloroform and applied to zone 1 of the chromatographic plate, pre-activated at 80 degrees for 30
minutes. As a “witness”, 2-3 µl of a standard tetrahydrocannabinol solution is applied to zone 2.
B. The plate is placed in a chromatography chamber saturated with vapors of a mixture of solvents
of the system :
• Petroleum ether and diethyl ether are mixed in a ratio (4:1) The length of the solvent front path
is 10 cm B. Dried at room temperature or in a current of warm air until the odor of solvents is
completely removed. The plate is evenly sprayed with 0.5% solution of durable blue B in 10%
aqueous solution of sodium carbonate D. In the area of the chromatogram of the “witness”, red-
purple spots with Rf = 0.8 are observed, which indicates the presence of tetrahydrocannabinol
(reaction sensitivity is 0.1 micrograms).
Conclusion
This study addresses a critical gap in forensic science by providing a comprehensive guide to the
chemical detection of cannabinoids in biological fluids. The research highlights the effectiveness
of urine as a readily accessible and informative sample for cannabinoid analysis, emphasizing the
sensitivity and reliability of thin-layer chromatography (TLC) with a sensitivity of 0.1 micrograms
for achieving precise and conclusive results. The detailed procedures for isolating cannabinoids
from various biological fluids, including saliva, blood, urine, and skin swabs, provide valuable
resources for forensic practitioners and researchers. This study contributes significantly to
advancing forensic science by offering a practical and reliable approach for detecting cannabinoids
in different biological samples. Further research should explore the application of these methods
in various case scenarios and investigate the potential for developing more sensitive and automated
techniques for cannabinoid detection.
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INTERNATIONAL JOURNAL OF ARTIFICIAL INTELLIGENCE
ISSN: 2692-5206, Impact Factor: 12,23
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