• Animal cells and systems
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• v.3 no.2
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• pp.215-219
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• 1999

Monoclonal antibody for delta-9-tetrahydrocannabiol (THC Ab), conjugated with protein A-gold, was employed as a probe to detect THC localization in the gland and subjacent cells of chemically fixed bracts of Cannabis. THC was detected in the outer wall of the disc cells, fibrillar matrix, the surface feature of secretory vesicles, and sheath throughout development of the secretory cavity. The probe was absent from vesicles. Label was also present in anticlinal walls of disc cells and walls of dermal and mesophyll cells. Little or no THC Ab was present in disc cells and none were detected in control tissues. This distribution pattern of THC Ab was similar to that in tissues prepared by high pressure cryofixation-cryosubstitution. Consistent association of THC with wall and wall-derived materials suggests that cannnabinoids are synthesized outside the plasma membrane and bound to a wall component, where-upon they are transported to the cavity with wall materials released from the disc cell wall during development of the secretory cavity.

• Archives of Pharmacal Research
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• v.28 no.9
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• pp.1086-1091
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• 2005

An analytical method was developed for evaluating the cannabidiol (CBO), cannabinol (CBN), ${\Delta}^9-tetrahydrocannabinol$ $({\Delta}^9-THC)$ level in human hair using gas chromatography-mass spectrometry (GC-MS). Hair samples (50mg) were washed with isopropyl alcohol and cut into small fragments (< 1mm). After adding a deuterated internal standard, the hair samples were incubated in 1.0M NaOH for 10 min at $95^{\circ}C$. The analytes from the resulting hydrolyzed samples were extracted using a mixture of n-hexane-ethyl acetate (75:25, v/v). The extracts were then evaporated, derivatized, and injected into the GC-MS. The recovery ranges of CBD, CBN, and ${\Delta}^9-THC$ at three concentration levels were $37.9-94.5\%$ with good correlation coefficients $(r^2>0.9989)$. The intra-day precision and accuracy ranged from $-9.4\%\;to\;17.7\%$, and the inter-day precision and accuracy ranged from $-15.5\%\;to\;14.5\%$, respectively. The limits of detection (LOD) for CBD, CBN, and ${\Delta}^9-THC$ were 0.005, 0.002, and 0.006 ng/mg, respectively. The applicability of this method of analyzing the hair samples from cannabis abusers was demonstrated.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.121-121
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• 2022

마자인(麻子仁)은 뽕나무과에 속하는 대마(Cannabis sativa L.)의 종자로써, 治血虛津虧, 腸燥便秘하며, 中風汗出, 逐水, 利小便, 破積血, 復血脈, 乳婦産後餘疾, 長髮 등에 사용한다고 기록되어 있다. 마자인 종자의 외피에는 환각성분인 Tetrahydrocannabinol (THC) 및 뇌전증과 다발성경화증 등 희귀, 난치성 질환의 치료제로 사용되는 Cannabidiol (CBD) 성분이 함유되어 있다. 따라서 외피를 제거한 마자인은 햄프씨드(Hemp Seed)로써 식품으로 이용되지만, 외피를 제거하지 않은 마자인은 의약품으로써 사용이 되고 있다. 마자인은 윤조탕, 자윤환, 마자인환 등 다양한 한의학 처방에 사용되지만, 외피에 함유되어있는 THC 및 CBD의 함유량 및 안전성에 대한 연구는 밝혀지지 않았다. 이에 마자인 함유처방의 안전성을 입증하는 실험방법을 설계하였다. 우선 의약품으로 유통되는 마자인의 외피에 THC 및 CBD 성분이 있는지를 확인하기 위해 마자인을 다양한 용매별 (물, 헥산 등)로 추출한 후 LC/MS를 이용해 성분 유무를 확인한다. 또한, 마자인 함유 처방 (마자인환, 자윤환, 윤조탕 등)을 복용하였을 경우 혈중에 THC 및 CBD 성분이 있는지를 확인하는 방법으로 실험동물에 마자인 함유 /처방을 투여한 후 혈액을 채취한다. 마찬가지로 성분 유무를 측정하는 방법은 LC/MS를 이용해 확인한다.

• Journal of Life Science
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• v.31 no.7
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• pp.677-685
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• 2021

Cannabis sativa L. belongs to the Cannabaceae family and is an annual herbaceous flowing plant. The plants can be classified into narcotic marijuana and nonnarcotic hemp. Different parts of C. sativa L. have been used as food, medicine, cosmetics, fiber and textile. However, the use of leaf, flower, and seed of C. sativa L was forbidden in Korea in January 1977 as a result of the Cannabis Control Act due to the narcotic properties. The plant's mature stems have limited uses for the production of fiber and sheets. Recently, various cannabinoids, terpenes and essential fatty acids were identified from C. sativa L., and their safety and useful bio-activities, such as neuroprotective, anti-inflammation, antithrombosis, antiepileptic, and antimicrobial activities, and the relief of pain, have been highlighted. Furthermore, the process of reduction of tetrahydrocannabinol, a representative narcotic compound, and the isolation of cannabidiol, a nonnarcotic active compound in C. sativa L., have been determined. These findings resulted in the legalization of C. sativa L. in Korea for medical use in December 2018 and the exclusion of C. sativa L. from the narcotic list of the UN Commission on Narcotics Drugs (UNCND) in December 2020. Therefore, developments of various high-value added products have commenced worldwide. Additionally, in 2021, the Korean government deregulated special zones based on hemp. In this study, the current status and the prospect of the hemp industry, as well as essential techniques for developing new hemp products, are provided for the activation of the Korea Green-Rush.

• Analytical Science and Technology
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• v.34 no.6
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• pp.241-250
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• 2021

Calibration curves are essential in quantitative methods and for improving the accuracy of analyte measurements in biological samples. In this study, a statistical analysis model built in the R language (The R Foundation for Statistical Computing) was used to identify a set of weighting factors and regression models based on a stepwise selection criteria. An LC-MS/MS method was used to detect the presence of urinary methamphetamine, amphetamine, and 11-nor-9-carboxy-Δ⁹ -tetrahydrocannabinol in a sample set. Weighting factors for the calibration curves were derived by calculating the heteroscedasticity of the measurements, where the presence of heteroscedasticity was determined via variance tests. The optimal regression model and weighting factor were chosen according to the sum of the absolute percentage relative error. Subsequently, the order of the regression model was calculated using a partial variance test. The proposed statistical analysis tool facilitated selection of the optimal calibration model and detection of methamphetamine, amphetamine, and 11-nor-9-carboxy-Δ⁹-tetrahydrocannabinol in urine. Thus, this study for the selection of weighting and the use of a complex regression equation may provide insights for linear and quadratic regressions in analytical and bioanalytical measurements.

• YAKHAK HOEJI
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• v.48 no.3
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• pp.207-212
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• 2004

An analytic method was developed for the quantitation of $\Delta$$^{9}-$ tetrahydrocannabinol (THC) and 11-nor-9-carboxy THC (THC-COOH) in human hair. After hair samples were pulverized using Freezer Mill, deuterated internal standards were added and digested in 1 N NaOH at $100^{\circ}C$ water bath for 30 min. Digest solutions were extracted by 5 ml hexane：ethyl acetate (90：10) after acidification with acetic acid. The organic phase was evaporated under N 2 and derivatized by BSTFA (with 1% TMCS) at $85^{\circ}C$ for 45 min. The derivatized solution was separated on HP-5MS column ($30m{\times}0.25mm{\times}0.25mm$) and detected using EI-GC-MS with selective ion monitoring mode. The assay of calibration was ranged from 5 to 100 ng/50 mg hair ($r^2$＞0.99) for THC and THC-COOH. Within and between-run precision were calculated at 6, 30, 60 ng/50 mg hair with coefficients of variation less than 11%. Within and between run accuracies at the same concentrations were$\pm$14% and $\pm$30% of target for both analytes, respectively. Absolute and relative recovery at 10 and 100 ng were 60∼91％. The method was used to detect and quantify THC and THC-COOH in cannabis abuser's hairs (N = 16) and SRM (N=5, THC 1 ng/mg, NIST). We detected THC and THC-COOH in only one hair sample. In SRM, % accuracy was 93% (range 86∼103%) and precision (% CV) was 8.14. We began to set up a quantitative analysis of THC and THC-COOH using EI-GC-MS. Continuously, we need to modify and develop this method in order to apply for identification in cannanbis users' hair.

• Analytical Science and Technology
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• v.34 no.2
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• pp.68-77
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• 2021

Cannabis is one of the most abused drugs in Korea. The main psychoactive component in cannabis, Δ9-tetrahydrocannabinol, is metabolized to 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) and THCCOOH-glucuronide (THCCOOH-glu) in the human liver, whereby the amount of THCCOOH-glu found in urine is twice as high as that of THCCOOH. The analytical process adapted by the majority of urine drug-testing programs involves a two-step method consisting of an initial immunoassay-based screening test followed by a confirmatory test if the screening test result is positive. In this study, a qualitative gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the detection of THCCOOH in human urine, where THCCOOH-glu was converted into THCCOOH by alkaline hydrolysis. For purification of the urine extract prior to instrumental analysis, high-speed centrifugation was used to minimize interference. In addition, an injection-port derivatization method using ethyl acetate and N,O-bis(trimethylsilyl)-trifluoroacetamide containing 1 % trimethylchlorosilane was employed to reduce the time required for derivatization, and an aliquot of the final solution was injected into the GC-MS. The method was validated by measuring the selectivity, limit of detection (LOD), and repeatability. The sensitivity, specificity, precision, accuracy, Kappa, F-measure, false positive, and false negative rate were determined by comparing the GC-MS results with those obtained using the immunoassay. The LOD was determined to be 0.32 ng/mL, while the repeatability was within 9.1 % for THCCOOH. Furthermore, a comparison study was carried out, whereby the screening immunoassay exhibited a sensitivity of 86.4 % and a specificity of 100 % compared to GC-MS. The applicability of the developed method was examined by analyzing spiked urine and forensic urine samples obtained from suspected cannabis abusers (n = 221).

• YAKHAK HOEJI
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• v.17 no.1
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• pp.21-26
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• 1973

The constituents of Korean cannabis were studied comparatively with foreign orgins. Especially, tetrahydrocannabinol(THC), the active constituents of cannabis were analyzed by the technique of thin layer chromatography and colorimetry. The results are as follows ; (1) THC content in Korean cannabis is comparatively higher than that in foreign samples. (2) THC is contained most abundantly in male flowers, abundantly in female tops and leaves and some in barks. (3) There is a tendency that the THC content increase gradually with growth of plants, being highest during unripe nad decrease with maturity of the tops. (4) Korean cannabis contains THC, cannabinol, cannabidiol, cannabidiolic acid and other cannandiolic compounds. Distribution of chemical components of Korean cannabis, compared with those of foreign ones, is remarkably different. (5) The THC content of Spanish cannabis cultivated tentatively in Korea is similar to that of Korean cannabis.

• Investigative Magnetic Resonance Imaging
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• v.22 no.3
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• pp.168-171
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• 2018

Cannabis or marijuana is the most commonly used recreational drug after alcohol in the world, and usage is generally recognized as having few serious adverse effects. However, usage is restricted in South Korea. The report of ischemic stroke associated with cannabis is rare in literature. We present a case of a 47-year-old female patient with no underlying disease presenting with acute ischemic stroke after smoking cannabis in South Korea. The result for synthetic cannabinoid metabolites (delta-9 tetrahydrocannabinol) screening was positive. Absence of other vascular risk factors and drug screening results suggest a causal role of cannabis in this ischemic stroke case. The patient eventually progressed to brain death. The underlying mechanism, clinical manifestation, and imaging findings of cannabis-related stroke will be reviewed.

• Toxicological Research
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• v.35 no.1
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• pp.37-44
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• 2019

A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.