氯胺酮及其代谢物去甲氯胺酮在大鼠体液及组织中的检测及分布

目的:建立生物检材中氯胺酮及其代谢物去甲氯胺酮的检测方法,考察氯胺酮和去甲氯胺酮在大鼠体内的分布。方法:用高效液相色谱法(HPLC)检测生物检材中的氯胺酮及去甲氯胺酮;以10 mg·kg-1剂量给大鼠灌胃(ig),分别于给药后10 min,45 min,2 h和8 h处死,测定体液和组织中氯胺酮及去甲氯胺酮的含量。结果:建立起生物检材中氯胺酮及其代谢物去甲氯胺酮的HPLC法。所建方法在0.05-25.0μg.g-1浓度范围内线性关系良好,r2=0.999,萃取回收率大于75%;肝和肾中氯胺酮和去甲氯胺酮含量最高,其次为:脾、尿液、肺、脑、血液、心和肌肉。结论:所建方法准确、灵敏,适用于组织中氯胺酮及其代谢物去甲氯胺酮的分析鉴定;肝和肾可作为毒物分析的组织检材。     

尿液中氯胺酮及其代谢物的气相-质谱分析

目的:建立尿液样品中氯胺酮及其代谢物去甲氯胺酮、去氢去甲氯胺酮、脱氢去甲去氨氯胺酮的快速提取及气相-质谱(GC-MS)分析方法。方法:尿液用10%碳酸钠调节pH=9～10,乙醚液液萃取,GC-MS分析。结果:该方法可快速检测氯胺酮滥用者尿液中的氯胺酮原体及三个代谢物,同时检测包括苯丙胺类药物在内的其它药物。结论:该方法可快速进行氯胺酮滥用者尿液的分析。     

尿中氯胺酮及其代谢物的定性分析

目的:了解氯胺酮在人体内的代谢情况和代谢产物,为检测滥用氯胺酮者尿液提供定性分析手段.方法:用固相萃取及GC/MS的选择性离子模式(SIM)检测尿中氯胺酮及其2种代谢物.结果:运用GC/MS确认了氯胺酮滥用者尿液中的有效成分氯胺铜及其代谢物去甲基氯胺酮和脱氢去甲基氯胺酮.结论:上述方法可以用于滥用者尿中氯胺酮及其代谢物的测定.     

氯胺酮类似物研究进展

近年来,包含氯胺酮类似物在内的新精神活性物质不断涌现,在世界范围内引起严重的卫生问题和社会问题。氯胺酮类似物以芳基环乙胺为母核,通过有针对性的结构修饰,达到与氯胺酮相似甚至更强劲的生理反应。过量服用氯胺酮类似物而导致急性死亡的案例被广泛报道,为了快速精准地控制和打击该类物质的滥用和犯罪,各国根据实际情况开发了各种筛查与定性定量方法。本文对氯胺酮类似物的化学结构、毒理学性质、中毒案例及近年来生物检材中的检测方法进行总结,并对氯胺酮类似物的毒理学研究提出了展望。     

用液相色谱-离子阱质谱联用法检测兔体液中阿普唑仑及其主要代谢产物

建立鉴别体液中阿普唑仑及其主要代谢物的高效液相色谱 电喷雾离子阱质谱联用(LC/MSn)法 .采用LC/MSn联用技术 ,检测家兔灌胃给药后血样和尿样中阿普唑仑原形药及其羟基化代谢物、葡萄糖苷酸型结合物 ,得到了它们的色谱、质谱以及特征碎片离子信息 .阿普唑仑的最低检测限小于 1ng .本法专属性强 ,可推广至其他生物检材的测定 ,尤其适用于需快速响应的临床和法医毒物分析     

手印中残留氯胺酮成分的检测分析

目的:建立一种手印中残留氯胺酮成分的分析方法。方法:对非渗透性和渗透性两种检材分别提取,采用GC/MS定性和GC/FID定量的方法对粘附有氯胺酮的手印样本进行定性定量分析。结果:氯胺酮浓度在10～4000ng·μL-1之间具有良好的线性关系(r=0.9991),最低检出限为1.635ng·μL-1。结论:通过实验发现,对于不同检材上捺印的手印均能检测到的被测物,通过手印显现方法处理后仍能检出。但随着陈旧时间的递增,有部分未检出。此方法为测定手印残留氯胺酮成分的定量分析提供了分析方法和检测依据。     

薄层色谱扫描法和气相色谱/质谱法快速诊断有机磷农药中毒

目的建立薄层色谱法快速定性和气相色谱(GC)/质谱(MS)法定性定量检测有机磷农药,快速诊断有机磷农药中毒。方法检材经二氯甲烷萃取后,薄层色谱法快速定性,GC/MS选择离子模式定性定量检测检材中有机磷农药浓度。结果对硫磷、甲拌磷、马拉硫磷、辛硫磷和久效磷在紫外灯下均有较好的紫外吸收,用双波长薄层色谱扫描仪对斑点做原位紫外吸收扫描,最大吸收波长分别为300,240,225,290,260nm,可配合比移值进行定性分析;GC/MS选择离子检测方式进行定性定量检测,各药物的特征离子峰分别为:97,109,291;76,97,121;93,125,173;77,97,129;67,97,127。结论薄层色谱扫描法定性检测检材中有机磷农药,具有快速、准确、操作简便等特点,加上GC/MS定性定量检测,不仅提高了方法的灵敏度,而且使结果更加准确可靠,可应用于有机磷农药中毒的快速诊断和有机磷农药中毒致死的法医学鉴定。     

LC-MS/MS快速测定生物检材中乌头碱的含量

目的建立快速测定生物检材中乌头碱的LC-MS/MS方法。方法生物检材经乙醚-二氯甲烷（3：2）提取后,用Venusil Mp-C18柱进行分离,采用ESI多反应监测（MRM）扫描方式。结果测定的线性范围为3～5000ng·mL-1,RSD〈1.8%,日间和日内精密度分别为2.5%和4.6%,提取回收率为85.61%～89.04%。结论该方法简便、快捷,适用于生物检材中乌头碱的快速测定。     

8种不同检材提取DNA的结果分析

在对不同检材提取、扩增、检测DNA中，如何对样本快速、正确、高效地提取出高质量的DNA图谱结果是影响DNA检验质量和效率的关键。目前DNA提取的方法有多种，其中磁珠法与ehelex-100提取法在各实验室的DNA提取检测方法比较常见。通过反复比较各种检材DNA提取方法的易操作性和结果的稳定性，确立了以磁珠法提取DNA的方法为主，ehelex-100法为辅的一系列提取方法。通过对不同检材样本的分析，选择用不同方法提取DNA。针对样本的质地、质量、数量等原因的不同，选择不同方法提取DNA，对检验结果有实际指导意义，在最后分析阶段也有不同的效果。     

毛发中大麻类物质的检测

目的对毛发中大麻类物质的检测方法进行综述。方法参考国内外近期的有关文献，介绍了毛发中大麻类物质分析的基本理论及发展概况，从毛发样品的采集、预处理、分析方法、优势和不足等几方面概述了毛发分析的过程。结果毛发作为生物检材具有不易造假，可反映药物长期滥用的情况等特点。结论毛发作为生物检材具有广泛的应用前景。     

在干细胞药物临床前研究中动物样品基因组DNA高通量提取的固相萃取法和磁珠筛选法初探

干细胞药物临床前研究中,常需要以干细胞特定基因片段为靶点追踪其体内的分布过程,需要提取动物样品基因组DNA,涉及到心脏、肝脏、肺脏、脾脏、肾脏、脑、骨髓、血、脂肪、肌肉、性腺、胃和肠等多种组织类型。为实现临床前样品的高通量检测,总结动物组织基因组DNA提取方法,并就如何高效高质提取样品基因组DNA进行讨论,提出间充质干细胞临床前研究中样品处理的一种可行的方案：通过生物样品制备仪获取稳定均一的组织裂解液,利用固相萃取法和磁珠筛选法配合使用获得高质量高浓度的动物样品DNA。     

内源性物质药物的定量测定及生物等效性评价研究进展

从定量分析和方法验证的角度来看,生物样品中内源性化合物的定量测定是非常复杂和困难的,而内源性药物的生物等效性评价同样也有其特殊性。文中通过对国内外文献资料进行调研、分析和总结,详细综述了内源性药物的生物分析方法及生物等效性试验设计和评价方法,并对面临的不同问题提出针对性解决方法。本文提供的一些思路和方法,对内源性药物的生物样品分析及生物等效性评价有重要意义。     

Urinary excretion rates of ketamine and norketamine following therapeutic ketamine administration: method and detection window considerations

Ketamine is widely used in veterinary medicine. Its medical application in humans is limited to children because in adults it induces severe psychedelic episodes. In recent years, teenagers have abused ketamine as a recreational and "club drug" because of its hallucinogenic and stimulant effects. Ketamine is also misused as a "date-rape" drug (to induce amnesia in unsuspecting victims). Sensitive gas chromatography-mass spectrometry-negative chemical ionization (GC-MS-NCI) and liquid chromatography-mass spectrometry-atmospheric pressure chemical ionization (LC-MS-APCI) methods were applied for the simultaneous quantification of ketamine and its major metabolite, norketamine, in urine. Urine samples were collected from hospitalized children who had received ketamine as an anesthetic. Individual urine samples were collected up to 16 days after drug administration. Using the GC-MS-NCI method, ketamine was detected in the urine of the children from only the day of drug administration up to 2 days after drug administration. Its concentrations ranged from 29 to 1410 ng/mL. Norketamine (measured in concentrations of 0.1-1442 ng/mL) was detected up to 14 days. Using the LC-MS-APCI method, norketamine was detected up to 6 days after drug administration, ranging in concentrations of 2-1559 ng/mL, while ketamine was detected up to 11 days (2-1204 ng/mL). In the urine taken from one child, ketamine was not detected through the entire 16-day period using both methods. The detection window for the analytes is highly dependent on the method used for determination and varies between individuals.     

生物样本中季铵类除草剂检测的研究进展

为精确检测生物样本中的百草枯、敌草快等季铵类除草剂提供参考，查阅国内外文献，对百草枯及敌草快的生物样本检测方法进行归纳和综述。结果显示，临床较为常用的检测方法为液相色谱法和液质联用法，受到检测方法、经济条件等因素影响。开展此类毒物检测的医院较少。生物样本中季铵类除草剂的检测方法仍需进一步的改进和推广，以更好地辅助临床诊断和治疗。     

Immunoaffinity Extraction and Alternative Approaches for the Analysis of Toxins in Environmental,Food or Biological Matrices

The evolution of instrumentation in terms of separation and detection allowed a real improvement of the sensitivity and analysis time. However, the analysis of ultra-traces of toxins in complex samples requires often a step of purification and even preconcentration before their chromatographic analysis. Therefore, immunoaffinity sorbents based on specific antibodies thus providing a molecular recognition mechanism appear as powerful tools for the selective extraction of a target molecule and its structural analogs to obtain more reliable and sensitive quantitative analysis in environmental, food or biological matrices. This review focuses on immunosorbents that have proven their efficiency in selectively extracting various types of toxins of various sizes (from small mycotoxins to large proteins) and physicochemical properties. Immunosorbents are now commercially available, and their use has been validated for numerous applications. The wide variety of samples to be analyzed, as well as extraction conditions and their impact on extraction yields, is discussed. In addition, their potential for purification and thus suppression of matrix effects, responsible for quantification problems especially in mass spectrometry, is presented. Due to their similar properties, molecularly imprinted polymers and aptamer-based sorbents that appear to be an interesting alternative to antibodies are also briefly addressed by comparing their potential with that of immunosorbents.     

The role of ketamine on plasma cocaine pharmacokinetics in rat

Ketamine has gained attention recently because of re-emergence of its abuse especially in combination with cocaine. When more than one drug is present simultaneously, the potential for drug--drug interaction exists, which can be pharmacokinetic, pharmacodynamic or both in nature. The objective of this study was to investigate the effect of ketamine on plasma cocaine pharmacokinetics to assess the role that the kinetic component may play in the interaction of these agents. Moreover, the effect of repetitive administration of ketamine pretreatment on the pharmacokinetics of cocaine was addressed. Male Sprague-Dawley rats were treated with cocaine alone (5 mg/kg i.v.), ketamine alone (100 mg/kg by gavage), or ketamine followed by cocaine (the same routes and doses). Blood samples were withdrawn at different time points post-injection and analyzed for determination of cocaine, its metabolites (benzoylecgonine and norcocaine) and ketamine. The results demonstrated that ketamine caused a significant decrease in cocaine's area under the curve (AUC) and elimination half-life while its total clearance was increased. The AUC of benzoylecgonine was increased by 1.5-fold after the combination compared with cocaine alone. However, cocaine did not affect ketamine's pharmacokinetic parameters. In the pretreatment study, ketamine was given orally for 3 days, followed 18 h later by a single i.v. of cocaine. Further enhancement of cocaine metabolism occurred with the appearance of norcocaine. This investigation revealed that ketamine enhances cocaine metabolism and may affect its toxicological profile.     

A novel micro-extraction strategy for extraction of bisphosphonates from biological fluids using zirconia nanoparticles coupled with spectrofluorimetry and high performance liquid chromatography

Extraction of bisphosphonates from biological fluids is important and time consuming step in sample preparation procedure. This paper describes a simple and green sample preparation technique for dispersive micro solid phase extraction (DMSPE) of alendronate sodium (ALS) fromurine and serum samples prior to direct spectrofluorimetry (DSFL) and high performance liquid chromatography with fluorescence detection (HPLC-FLD), respectively. The DMSPE strategy is based on the selective chemisorption of ALS on zirconia nanoparticles (ZNPs) as an adsorbent followed by derivatization of the eluted analyte using o-phthalaldehyde (OPA) in the presence of 2-mercaptoethanol (2ME) at basic medium to form fluorescent species. The chemical and instrumental influencing parameters on DMSPE and measuring methods were optimized for the efficient extraction and determination of ALS. The presented methods were capable of extracting ALS from human urine and serum samples and determining over the wide ranges of 5–1000 and 5–2500 μg L⁻¹ with limits of detection (LOD) of 1.5 and 1.4 μg L⁻¹ for DSFL and HPLC methods, respectively. The relative recoveries for the three spiked standard levels of ALS in urine and serum samples ranged from 89.0% to 107.0%, and the intra-day relative standard deviations (%RSDs) were in the range of 2.9–7.9%.     

Exosomes in cancer immunoediting and immunotherapy

As an important means of communication among cells, exosomes are being studied more and more widely, especially in the context of cancer immunotherapy. In the phase of tumor immunoediting, exosomes derived from tumor cells and different immune cells have complex and changeable physiological functions, because they carry different proteins and nucleic acid from the source cells. Based on the role of exosomes in the communication between different cells, cancer treatment methods are also under continuous research. This review briefly introduces the molecular composition of exosomes, which is closely related to their secretion mechanism. Subsequently, the role of exosomes encapsulating different information molecules is summarized. The role of exosomes in the three phases of tumor immunoediting is introduced in detail, and the relevant literature of exosomes in the tumor immune microenvironment is summarized by using a novel framework for extracting relevant documents. Finally, it summarizes the various exosome-based immunotherapies currently proposed, as well as the challenges and future prospects of exosomes in tumor immunotherapy.     

An infrared spectroscopic approach to characterise white powders,easily applicable in the context of drug checking,drug prevention and on-site analysis

More and more events, such as the summer music festivals, are considering the possibilities for implementing on-site testing of psychoactive drugs in the context of prevention and harm reduction. Although the on-site identification is already implemented by plenty of drug checking services, the required rapid quantitative dosing of the composition of illicit substances is still a missing aspect for a successful harm reduction strategy at events. In this paper, an approach is presented to identify white powders as amphetamine, cocaine, ketamine or others and to estimate the purity of the amphetamine, cocaine and ketamine samples using spectroscopic techniques hyphenated with partial least squares (PLS) modelling. For identification purposes, it was observed that mid-infrared spectroscopy hyphenated with PLS-discriminant analysis allowed the distinction between amphetamine, cocaine, ketamine and other samples and this with a correct classification rate of 93.1% for an external test set. For quantitative estimation, near-infrared spectroscopy was more performant and allowed the estimation of the dosage/purity of the amphetamine, cocaine and ketamine samples with an error of more or less 10% w/w. An easily applicable, practical and cost-effective approach for on-site characterisation of the majority of the psychoactive samples encountered in Belgian nightlife settings based on IR spectroscopy was proposed.