生物检材中氯胺酮及其代谢物的分析方法研究进展

综述了生物检材中氯胺酮及其代谢物分析技术的进展,评述了液相萃取、固相萃取、固相微萃取、顶空固相微萃取、微波辅助萃取等技术在分析中的应用,以及薄层色谱、气相色谱、高效液相色谱、毛细管电泳及其联用检测技术、酶联免疫分析技术和太赫兹时域光谱等检测技术在分析生物检材中氯胺酮的优势和局限性,并展望了该领域研究的发展趋势.     

可卡因免疫及适配体生物传感器

由于长期滥用可卡因会对人体产生心律失常、心肌梗死、中风、高血压、主动脉僵硬等不良影响,可卡因已成为当今最危险和非法滥用的药物之一,传统的可卡因色谱分析方法存在耗时、样本处理繁琐和操作复杂等缺点。因此,改善传统可卡因分析方法对打击犯罪和发展医学具有一定的积极影响。由于生物传感器的准确性和便携性,基于免疫和适配体技术的生物传感器是检测可卡因的一个重要发展方向。在这篇综述中,主要讲述了近年来不同类型的可卡因生物传感器,涵盖了基于电化学、荧光、比色等方法在可卡因检测上的进展,对可卡因的免疫和适配体生物传感器进行了归纳和综述,并总结了可卡因传感器的优缺点和发展方向。     

基于核酸适配子识别的荧光法检测可卡因

基于荧光标记和核酸适配子识别可卡因,建立了简单、灵敏的可卡因新型荧光分析法.在微孔板表面组装亲和素-生物素化可卡因适配子-FAM标记可卡因适配子互补短链复合物,根据加入可卡因前后荧光强度的变化来定量可卡因.实验考察了微孔板包被亲和素浓度、生物素标记适配子用量、FAM标记可卡因适配子互补短链用量、反应温度、反应时间等因素...     

气相色谱法测定尿液中可卡因及其代谢物爱冈宁甲基酯

建立了尿样中可卡因（COC）及其代谢物爱冈宁甲基酯（EME）的气相色谱检测方法。 采用液液萃取法提取尿样中可卡因和爱冈宁甲基酯,考察了萃取剂种类和用量、试样pH值以及萃取时间等因素对提取效果的影响。 结果表明,尿样中COC和EME的最佳液液萃取条件是：以V(氯仿)∶V(异丙醇)=9∶1为提取溶剂,调节样品溶液pH=9.5,在40 ℃ 水浴振荡提取6 min。 COC和EME日内精密度分别为1.73%和1.44%,日间精密度分别为2.57%和2.89%,最低检出限（LOD）为0.040 mg/L。 此法无需衍生化、快速、准确、灵敏度高,可同时检测尿样中COC和EME的含量。     

基于磁性微球分离技术的核酸适体化学发光检测可卡因

设计了一种基于磁性微球与核酸适体的夹心式化学发光适体传感器,建立了高灵敏度的可卡因分析方法。实验考察了反应所用羧基磁性微球、捕获探针、可卡因适体、生物素标记的报告序列以及链霉亲合素修饰的辣根过氧化物酶用量对化学发光信号的影响。优化条件下,在1.0×10-8~1.0×10-4mol/L范围内,化学发光信号与可卡因浓度的对数呈线性相关(r2=0.989 7),检出限为3.2×10-9mol/L。考察了共存物质中适体对可卡因的特异性识别能力,方法显示了较好的选择性。     

微波萃取-气相色谱法测定血液中的可卡因及其代谢物爱冈宁甲基酯

建立了血液中可卡因(cocaine, COC)及其代谢物爱冈宁甲基酯(ecgonine methyl ester, EME)的气相色谱-质谱(CG-MS)和气相色谱-氢火焰离子化检测(GC-FID)方法。该方法采用微波萃取提取血液中的COC和EME,优化并确定了最佳提取条件: 以氯仿-异丙醇(体积比为9:1)混合溶液为提取溶剂,用0.05 mol/L Na2CO3-NaHCO3缓冲溶液调节样品溶液的pH至10.0,在40 ℃下微波萃取6 min;采用GC-MS对萃取液中的COC和EME进行定性,采用GC-FID进行定量检测。COC和EME的平均回收率分别为79.91%～99.85%,相对标准偏差(RSD)均小于3.10%,检出限(S/N=3)分别为60 mg/L和40 mg/L。该方法无需衍生化,快速、准确、灵敏,可同时检测血液中的COC和EME。     

硝基呋喃类代谢物残留量检测标准的比较

对我国现有硝基呋喃类代谢物检测标准进行比较和分析,同时用不同检测标准对鳗鱼、虾、猪肉、牙鲆鱼、白鲫鱼等阳性样品的测定结果进行了统计比较分析.结果表明:各检测方法中,pH在7.0～7.5范围内对硝基呋喃类代谢物的乙酸乙酯萃取影响不大;用t -检验:双样本等方差假设比较两种方法的差异显著性表明,由于样品制备中洗涤和不洗涤的...     

稀土上转换发光纳米探针用于潜指纹中可卡因的检测

结合核酸适配体和稀土掺杂上转换纳米材料的优势,建立了一种潜指纹中外源性物质检测的新方法。Zeta电位、紫外吸收光谱和发光光谱表明核酸适配体成功修饰到上转换颗粒表面而且没有影响颗粒的发光性质。对含有不同量可卡因的指纹进行检测,在指纹中可卡因含量为0.5μg时仍能呈现出较强的发光强度。该方法能够通过潜指纹成像提供光学信号,实现潜指纹中外源性物质的检测,而且适用于不同人和不同基底上指纹的检测,具有简便、检测灵敏度高、适用性广等优点,对身份认证、刑事侦查和医疗诊断有重要意义。     

LC Method for the Determination of NPC1161, Primaquine and their Metabolites in Various Biological Systems

NPC1161 is an 8-aminoquinoline anti-malarial analog, which has a favorable toxicity profile relative to primaquine and other 8-aminoquinolines. High-performance liquid chromatographic method was developed for the determination of NPC1161, primaquine and their metabolites in biological samples in order to facilitate metabolic and pharmacokinetic studies. The method includes extraction of the unchanged drugs and their metabolites from the biological samples. Separation was achieved by reversed-phase chromatography on a C18 column with water–acetonitrile both containing 0.025% trifluoroacetic acid as the mobile phase. Recoveries of NPC1161 and its metabolites were greater than 60% in various biological samples tested. No interference with the components of the biological material was observed. The detector response was linear with concentrations of NPC1161 and its metabolites (desalkyl NPC1161 and carboxy NPC1161) in the ranges from 0.5 to 80.0, 0.4–60.0 and 0.4–70.0 μg mL⁻¹, respectively. A mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification of NPC1161 and its metabolites in biological samples. This method involved the use of the [M + H]⁺ions of NPC1161, C3 analog (internal std. for the assay), desalkyl NPC1161 and carboxy NPC1161 at m/z 434, 406, 349 and 449 in the positive ion mode with extractive ion monitoring (EIM). This method will have an important application in pharmacokinetic studies of NPC1161 and in understanding the mechanism of metabolism of this novel 8-aminoquinoline analog in more detail.     

Determination of Four Nitrofuran Metabolites and Chloramphenicolin Biological Samples Using Enzyme-Linked Immunosorbent Assay

A pretreatment method was developed for the determination of four nitrofuran metabolites and chloramphenicol in pork, chicken, fish, and shrimp. Homogenized samples were hydrolyzed and derivatized with 2-nitrobenzaldehyde. Extraction was performed using ethyl acetate followed by purification of the extract by hexane. Lastly, the ethyl acetate was dried under nitrogen and the residue was redissolved for analysis. The performance of the method was satisfactory for all drugs tested at contamination levels close to or below the relevant European Union maximum levels permitted. The limits of detection (LODs) of the method were 0.025–0.13 ng/g. Recoveries higher than 72.0% were obtained for all drugs tested, and the coefficient of variation was less than 15%. Results from analysis of unknown samples by the developed ELISA were similar to those obtained by a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.     

High-Performance Liquid Chromatographic Assay for Chlorquine and its Two Major Metabolites,Desethylchloroquine and Bidesethylchloroquine in Biological Fluids

Abstract

A high-performance liquid chromatography (HPLC) method for the determination of chloroquine and its two major metabolites in biological fluids is described. Hydroxychloroquine is used as an internal standard (I.S.). Drug, metabolites and I.S. were extracted as bases with diethyl ether by a single step procedure. After drying and evaporation of the organic phase, the residue was dissolved into the mobile phase and injected into the chromatographic system. Separation was performed using a normal phase column (Inertsil sill with mixture of acetonitrile, methanol and ammonia as mobile phase. The detection was carried out by fluorescence measurement : excitation wavelength was set at 325 nm and emission at 380 nm. The limit of detection was near 3.7 ng ml^?1 for chloroquine and metabolites. No chromatographic interference could be detected by endogenous compounds or other antimalarial drugs. Because of the good accuracy of the method, concentrations were determinated with a relative standard deviation lower than 7% at the 25 ng ml^?l level for all substances.

An excellent precision was obtained over the range of concentrations tested, 25–1000ng ml^?l. This method can be applied to therapeutic, pharmacokinetic and epidemial studies.     

Detection of Cocaine Using the Flow Immunosensor

Abstract

A continuous flow immunosensor has been designed for the detection of cocaine in aqueous samples. The continuous flow immunosensor relies on the displacement of fluorophore-labeled antigen from immobilized monoclonal antibody. The sensitivity and accuracy of the flow immunosensor were investigated while varying the parameters of immobilized antibody density, flow rate, amount of antibody-coated Sepharose used in each column, and the saturation of antibody binding sites with fluorophore-labeled antigen. Using a low density of immobilized anti-benzoylecgonine antibody, as little as 5 ng/ml cocaine could be detected. Small amounts of antibody-coated Sepharose could be used repeatedly and the lifetime of the column was proportional to the amount of Sepharose used. Results were obtained in less than a minute and cross-reactivity against various other drugs was negligible.     

Determination of Triazine Herbicides and Metabolites by Solid Phase Extraction with HPLC Analysis

An efficient solid phase extractive preconcentration/separation method was developed for the trace determination of herbicides in aqueous samples using Amberlite XAD-4 resin as the adsorbent. The retained herbicides were eluted with methanol at a flow rate of 1.0 mL min^?1 and determined by HPLC-DAD (wavelength of 220 nm) using water (pH:4.7, phosphoric acid) and methanol (ratio 35:65) as the mobile phase with a flow rate of 1.0 mL min^?1. Quantitative recoveries of simazine, atrazine and its metabolities were achieved at optimized analysis conditions that included 0.75 g of resin; a pH of 3.0; an eluent volume of 3.0 mL; an eluent flow rate of 1.0 mL min^?1; and a sample flow rate of 4.0 mL min^?1. The limits of detection, preconcentration factor, and linear ranges for the herbicides were 0.084–0.121 µgL^?1, 1000, and 0.5–20 mg L^?1, respectively. The performance of the method was evaluated by analysis of spiked water samples. The recoveries of simazine, atrazine and their metabolities were found to be quantitative (99.6–104.8%) with RSDs of 2.2–4.8% and 2.8–4.7% for intra-day and inter-day precision, respectively. The proposed method was successfully applied for trace determination of studied analytes in waste water, apple juice, and red wine samples.     

气相色谱-质谱法分析乳腺癌患者血清中的代谢物

建立了气相色谱-质谱法分析乳腺癌患者血清中代谢物的分析方法。分别收集乳腺癌患者、一般疾病者和健康对照组血清,样本用甲醇∶乙腈∶丙酮=1∶1∶1(V/V)提取,硅烷化试剂N,O-双甲氧基三氟乙酰胺+1%三甲基氯硅烷(BSTFA+1%TMCS)衍生后,采用气相色谱-质谱法分析其中的氨基酸、脂类和糖类的代谢谱,并用主成分分析(PCA)和随机森林(RF)算法对实验结果进行分析。结果表明,三组实验数据被成功分类,并发现磷酸、N-巴豆酰基甘氨酸、2,4-二羟基丁酸、棕榈酸、N-苯基甘氨酸和N-1-己酰甘氨酸等组分在乳腺癌组和健康对照组中的差异较为显著。     

人尿中避孕药炔诺酮代谢物的GC—MS分析

本文用GC－MS方法分析炔诺酮及其在人尿中的代谢产物，从收集的服药后5h的尿样中检出并鉴定了炔诺酮的2个主要物Met1和Met2，同时还发现22个浓度较低的次要代谢物Met3和Met4与诺龙的代谢情况不同，容易区别。     

Electrochemically Reduced Graphene Oxide for Forensic Electrochemistry: Detection of Cocaine and its Adulterants Paracetamol,Caffeine and Levamisole

This communication shows the improved performance of electrochemically‐reduced graphene oxide (ERGO) modified electrodes for the square‐wave voltammetric detection of cocaine (COC) and its main adulterants paracetamol (PAR), caffeine (CAF) and levamisole (LEV). The four compounds can be identified in a single voltammetric scan on the ERGO‐modified electrode with adequate resolution of peaks and increase in current (at least 2‐fold) in comparison with the unmodified electrode. Moreover, electrode fouling typically verified during the oxidation of CAF and LEV is eliminated. CAF and PAR were determined in a real COC sample and satisfactory recovery values were obtained (103 and 85 %, respectively).     

人体尿液中青霉素G代谢物的确认

建立了利用电场轨道阱回旋共振组合质谱仪(LTQ-Orbitrap)检测人体尿样中青霉素G及其代谢产物的分析方法。目标化合物分离鉴定选用PLRP-S聚合色谱柱,以含0.1%(体积比)甲酸溶液-乙腈为流动相,梯度洗脱分离后,选用Data-dependent MSn模式进行全扫描分析。尿样中首次检出青霉素甲酰化代谢产物(m/z 381)。青霉素新代谢物的发现对食品安全及法医毒物分析具有重要意义。