• 약학회지
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• 제54권4호
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• pp.226-231
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• 2010

Oral fluid has become increasingly popular as an alternative specimen in the field of driving under the influence of drugs (DUID) and work place drug testing. In this study, an analytical method for the detection and quantification of ${\Delta}^9$-tetrahydrocannabinol (THC) and its metabolite, 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THC-COOH) in oral fluid by SPE and GC-MS was established and fully validated. The stability of THC and THC-COOH in oral fluid during storage was also determined by examining the THC and THC-COOH concentration changes depending on time and container materials. Oral fluid samples were kept over 21 days at room temperature, $-4^{\circ}C$ and $-20^{\circ}C$ in two different specimen collection tubes; glass and polypropylene tubes. Three replicates for each condition with different temperature and types of a container were analyzed at five different time points over 21 days. When oral fluid samples were stored in glass tubes, the loss of both THC and THC-COOH was less than 10% at all room temperature, $-4^{\circ}C$ and $-20^{\circ}C$. However, in polypropylene tubes, the loss of both THC and THC-COOH increased significantly over the study period. In particular, the concentration of THC decreased more rapidly than that of THC-COOH at room temperature and the maximal percentage of THC lost was 90.3% after 21 days. The result indicates that it would be necessary to collect oral fluid samples in glass containers and cool the samples until analysis in order to prevent the degradation of analytes.

• Animal cells and systems
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• 제3권2호
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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.

• Bulletin of the Korean Chemical Society
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• 제12권6호
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• pp.604-606
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• 1991

From O-glucuronyl trichloroacetimidate (4) and ${\Delta}^6$-tetrahydrocannabinol (5), the C-glucuronide of ${\Delta}^6-tetrahydrocannabinol({\Delta}^6-THC)$ (7) was obtained using mild Lewis acid catalysis. The known method for the synthesis of C-glucuronide from 1-O-unprotected glucuronide provided low chemical yields. The C-glucuronide of ${\Delta}^6-THC$ was obtained through inversion of configuration at the anomeric center in O-glucuronyl trichloroacetimidate (4).

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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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.

• 약학회지
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• 제48권3호
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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.

• 분석과학
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• 제24권1호
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• pp.1-9
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• 2011

대마 흡연 여부 확인을 위해 사용되는 모발 중 대마 대사체 정량분석 결과의 측정불확도를 평가하였다. 대마초 성분 중 활성물질은 ${\Delta}^9$-tetrahydrocannabinol이며 대사과정을 거쳐 생성된 주요 대사체는 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THC-COOH)이다. 따라서 생체 대사물질인 THC-COOH는 대마 흡연 여부를 판단하는 지표물질로 사용되고 있다. 실험방법은 모발의 세척, 건조, 질량측정, 세절, 가수분해, 비드를 이용한 액체-액체 추출, 유도체화 반응 및 기기분석 과정으로 구성되었다. 측정의 소급성이 유지될 수 있도록 상위 규정기로부터 하위 측정기까지 교정을 통해 확보하였다. 측정불확도 평가에 앞서 정량분석시 측정값에 영향을 주는 인자들을 찾아내고 각각의 요소들이 측정결과에 어떤 영향을 주는가를 살펴보았다. 산출 결과, 재현성, 회수율, 검정곡선, 표준물질, 질량측정의 요소 순으로 불확도에 영향을 미치고 있음을 확인하였고 재현성의 요소가 측정불확도에 가장 큰 영향을 미치고 있었다. 실제 대마 복용자의 모발을 분석한 결과 대마 대사체 농도 측정값에 대한 오차 범위의 상대불확도는 17% 이였으며, 본 연구의 측정불확도 평가 결과는 향후 분석방법의 개선 및 측정결과의 신뢰도 제고를 위한 근거자료로 활용할 예정이다.

• 약학회지
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• 제17권1호
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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.

• Archives of Pharmacal Research
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• 제28권9호
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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.

• 약학회지
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• 제52권3호
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• pp.172-175
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• 2008

Stable isotope ratio of carbon and nitrogen ($\delta^{13}C$ & $\delta^{15}N$), and $\Delta^{9}$-tetrahydrocannabinol (THC) contents were measured on 37 Korean cannabis and 10 commercial grade marijuana seized in Korea. Factors influencing on the measured values and their variations were investigated. $\delta^{13}C$ value of cannabis is specified mainly by water availability. Korean cannabis showed relatively low $\delta^{13}C$ values ranging -33.29$\sim$-27.01% (mean=-31.01%), which reflect geographic conditions of Korea where is rainy, especially during summer. $\delta^{15}N$ values, which reflect individual planting conditions, were relatively high up to -0.5$\sim$18.0% (mean=6.44%). It reflects characteristics of Korean cannabis growing wild in forest or cultivated in fertile soil. Tetrahydrocannabinol is the major hallucinogenic compound of cannabis. Ethanol extracts of cannabis leaves were derivatized by N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), and the derivatives were analyzed by GC-MS in selected ion monitoring (SIM) mode. THC contents of Korean cannabis ranged 0.11$\sim$4.34% (mean=1.47%), which were relatively low compared with commercial grade marijuana.

• 분석과학
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• 제19권5호
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• pp.441-448
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• 2006

소변 중 대마 남용여부를 판별하는데 기준이 되는 tetrahydrocannabinol (THC)의 대사체 성분인 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THCCOOH)를 고체상 추출법 (solid-phase extraction, SPE)과 가스크로마토그래피/질량분석법 (GC/MS)을 이용하여 신속하게 분석 할 수 있는 방법을 제시하였다. 본 실험은 시험관에 소변 3 mL를 취해 염기성 (pH 10) 조건에서 가수분해 한 후, 양이온교환 카트리지를 사용하여 THCCOOH 성분을 선택적으로 추출하고, 증발 건고한 다음 유도체 반응을 시켜 GC/MS로 분석하였다. 그 결과 분석방법의 검출한계 (LOD)는 0.4 ng/mL이고, 정량한계 (LOQ)는 1.2 ng/mL이였다. 검정곡선의 직선성 상관계수 ($r^2$)는 1.2 (LLE는 1.3)~50.0 ng/mL의 농도범위에서 0.999를 나타내었다. 그리고 정밀도 (precision)와 정확도 (accuracy)는 모두 ${\pm}1.20%$ 이내로 안정적이었으며, 회수율(recovery)은 83.6~90.7%로 측정되었다. 액체상 추출법 (liquid-liquid extraction, LLE)과 비교할 때, SPE 방법이 회수율은 낮았지만 검출한계, 정량한계, 정밀도 및 정확도에서는 큰 차이가 없었다. 그러나 LLE 방법은 추출과정에 시간과 노력이 많이 드는 반면, SPE 방법은 상대적으로 추출 조작이 간편하고 신속하게 추출되었으며, 추출 잔류물도 깨끗하였다. SPE를 이용한 추출방법을 다수의 대마 흡연자 소변에 적용하였을 때 기존에 사용하던 LLE 방법보다 간편하고, 신속하게 대마 대사체 분석이 가능하였다.