云南大麻DNA的提取及检测初步研究

目的探讨云南大麻DNA的提取及检测方法。方法运用改进的SDS微量法提取大麻总DNA,对所得DNA进行PCR检测。结果用所筛选到的大麻引物检测了云南大麻的DNA遗传标记特征,图谱清晰,结果稳定。结论本方法能有效提取并检测大麻DNA,为实现大麻的品种鉴定、遗传多态性研究及来源追踪提供有价值的科学依据。     

Forensic laboratory backlog: The impact of inconclusive results of marijuana analysis and the implication on analytical routine

Forensic laboratories worldwide are struggling to keep up with the increasing number of cases submitted for analysis, regardless of the reasons, backlog of controlled substances cases is a reality in many countries. In this paper we analyse the number of petitioned examinations (from 2016 to 2020) and the data from 11,655 marijuana TLC results from the Forensic Laboratory in the Federal District Civil Police in Brazil. Data demonstrates that backlog increases inconclusive results, with storage and light playing a crucial role in the process. Additionally we explored the repercussions of delayed forensic results for controlled substances and propose an approach to overcome waiting time in this context.     

反相HPLC法同时测定大麻植物中的三种有效成分

目的建立同时测定大麻植物中四氢大麻酚(tetrahydrocannabinol,THC)、大麻二酚(cannabidiol,CBD)和大麻酚(cannabinol,CBN)三种有效成分的高效液相色谱(HPLC)法。方法采用通用C_(18)色谱柱,以乙腈-磷酸盐缓冲液(0.015 mol/L KH_2PO_4)为流动相,流速为1.0 m L/min,检测波长为220 nm,同时收集波长190~400nm的紫外光谱图,并以此光谱图及保留时间作为定性依据。结果所建方法能良好地分离THC、CBD和CBN,三种成分在0.4~40μg/m L范围内线性关系良好(R~2≥0.999 3),回收率大于87%,最低检出限分别为1.8、2.0和1.3 ng,日内精密度与日间精密度均小于5%。结论反相HPLC法简便、快速、准确,适用于大麻植物中THC、CBD和CBN的定性定量检测。     

Quantitative Determination of Cannabinoids in Cannabis and Cannabis Products Using Ultra‐High‐Performance Supercritical Fluid Chromatography and Diode Array/Mass Spectrometric Detection

Ultra‐high‐performance supercritical fluid chromatography (UHPSFC ) is an efficient analytical technique and has not been fully employed for the analysis of cannabis. Here, a novel method was developed for the analysis of 30 cannabis plant extracts and preparations using UHPSFC /PDA ‐MS . Nine of the most abundant cannabinoids, viz . CBD , ?⁸‐THC , THCV , ?⁹‐THC , CBN , CBG , THCA ‐A, CBDA , and CBGA , were quantitatively determined (RSD s < 6.9%). Unlike GC methods, no derivatization or decarboxylation was required prior to UHPSFC analysis. The UHPSFC chromatographic separation of cannabinoids displayed an inverse elution order compared to UHPLC . Combining with PDA ‐MS , this orthogonality is valuable for discrimination of cannabinoids in complex matrices. The developed method was validated, and the quantification results were compared with a standard UHPLC method. The RSD s of these two methods were within ±13.0%. Finally, chemometric analysis including principal component analysis (PCA ) and partial least squares‐discriminant analysis (PLS ‐DA ) were used to differentiate between cannabis samples.     

Field testing of collection cards for Cannabis sativa samples with a single hexanucleotide DNA marker

The validity and feasibility of using DNA collection cards in the field for preservation and analysis of Cannabis sativa genotypes were investigated using a highly specific hexanucleotide marker. Collection cards were submitted to the National Marijuana Initiative, which selectively trained and managed the collection of specific types of samples from a variety of participating agencies. Samples collected at seizure sites included fresh marijuana leaf samples, dried "dispensary" samples, U.S. border seizures, and hashish. Using a standardized PCR kit with custom-labeled oligonucleotide primers specific to marijuana, collection cards produced eight genotypes and 13 different alleles, extremely low baselines, and no cross-reactivity with control plant species. Results were produced from all sample types with the exception of hashish. Plant DNA collection cards represent an easily implementable method for the genetic identification and relatedness of C. sativa street and grow site-seized samples with applications for databasing and market disruption.     

大麻遗传标记的法医学应用研究进展

大麻是大麻科大麻属一年生雌雄异株的草本植物,其内含有具有强烈成瘾性和麻醉性的四氢大麻酚(THC).大麻价格低廉、获取方便、且受到一些国家和地区合法化的影响,目前已成为滥用最广泛的毒品之一.因此,大麻植株的鉴定对于打击毒品犯罪、维护社会稳定具有重要意义.近年来,基于DNA遗传标记的大麻鉴定为案件侦破提供了新的技术手段,针...     

Stable Isotope Ratios of Marijuana. I. Carbon and Nitrogen Stable Isotopes Describe Growth Conditions*

Abstract: There remains significant uncertainty in illicit marijuana cultivation. We analyzed the δ¹³C and δ¹⁵N of 508 domestic samples from known U.S.A. counties, 31 seized from a single location, 5 samples grown in Mexico and Colombia, and 10 northwest border seizures. For a subset, inflorescences and leaves were analyzed separately. These data revealed a strong correspondence, with inflorescences having slightly higher δ¹³C and δ¹⁵N values than leaves. A framework for interpreting these results is introduced and evaluated. Samples identified as outdoor‐grown by δ¹³C were generally recorded as such by the Drug Enforcement Administration (DEA). DEA‐classified indoor‐grown samples had the most negative δ¹³C values, consistent with indoor cultivation, although many were also in the outdoor‐grown domain. δ¹⁵N indicated a wide range of fertilizers across the dataset. Samples seized at the single location suggested multiple sources. Northwest border δ¹³C values suggested indoor growth, whereas for the Mexican and Colombian samples they indicated outdoor growth.     

Developmental validation of a Cannabis sativa STR multiplex system for forensic analysis

A developmental validation study based on recommendations of the Scientific Working Group on DNA Analysis Methods (SWGDAM) was conducted on a multiplex system of 10 Cannabis sativa short tandem repeat loci. Amplification of the loci in four multiplex reactions was tested across DNA from dried root, stem, and leaf sources, and DNA from fresh, frozen, and dried leaf tissue with a template DNA range of 10.0-0.01 ng. The loci were amplified and scored consistently for all DNA sources when DNA template was in the range of 10.0-1.0 ng. Some allelic dropout and PCR failure occurred in reactions with lower template DNA amounts. Overall, amplification was best using 10.0 ng of template DNA from dried leaf tissue indicating that this is the optimal source material. Cross species amplification was observed in Humulus lupulus for three loci but there was no allelic overlap. This is the first study following SWGDAM validation guidelines to validate short tandem repeat markers for forensic use in plants.     

应用AFLP检测大麻遗传多样性

目的 筛选对于大麻多态性好的AFLP引物标记来区分大麻品种.方法 利用AFLP技术用55对引物组合对12个大麻地域品种进行初筛.结果 选出5对多态性好的引物组合进行了遗传多样性研究.每对AFLP引物组合扩增出47～76务带.共获得285条带,其中多态性条带为99条以及10条品种特异带.结论 AFLP对于大麻具有很高的分辨率,为今后深入地研究大麻植物遗传多样性奠定了很好的基础.     

大麻研究现状综述

大庥是大麻科（cannabinaceae）大麻属（Cannabis）一年生草本植物,因其价格低廉、摄取方便,为目前使用最广泛的毒品之一。国内外专家学者对大麻的化学成分、分子生物学、药理作用等几个方面进行了广泛研究,并取得诸多进展,今后应加强对大麻背景知识的研究,ITS技术应用于大麻的研究。