Research of the extraction method of morphine from biological fluids

A solid-phase extraction method of morphine from urine and blood has introduced. The effect of 5 SPE columns, 3 eluents and pH on morphine recovery has been investigated systematically. Derivative GC was used as a method of detection. The result showed that the column and the eluent of such as GDX-301, GDX-403 and C18 chloroform:isopropanol (9:1) had good behaviors to extraction of morphine. When GDX-301 was used as a sorbent, the recovery of morphine from urine was above 90% at pH 9, then went down with the increase of pH. While the recovery from blood was growing with the increase of pH, which reached above 90% in strong alkaline. The extraction method is simple, inexpensive, efficient and reproducible, which provides an effective and practical method to extract morphine and similar illicit drugs from biological fluids.     

Detection of dipiridamol in biological fluids

Acetone is proposed as an isolating agent for dipiridamol isolation from biological fluids. Purification of the isolates was performed with liquid-liquid extraction and colon chromatography with silasorb C-18 sorbent. The technique of dipiridamol detection in the blood and urine is described. The assays results are presented.     

Detection of furadan in biological fluids

Optimal conditions have been determined for furadan isolation from biological fluids by means of the mixture of solvents ethyl acetate-acetone in volume 1:1. Possible purification of the compound from coextractive substances of biomaterial on the column with silica gel L 100/160 mcm is demonstrated. IR-spectrophotometric methods and high performace liquid chromatography with a detector of photodiode matrix are proposed for identification and quantitation of furadan in blood and urine specimens.     

A rapid method for the extraction of cocaine and benzoylecgonine from body fluids

A method has been developed for the rapid extraction of cocaine and benzoylecgonine from post-mortem blood and urine samples. Solid phase columns containing C8 packing material gave clean, emulsion free extracts from body fluids. The total time for each extraction was approx. 10 min and the system has the capacity for the simultaneous extraction of up to 10 samples. Urine can be extracted directly, blood samples were sonicated before extraction to allow for their easy passage through the columns. The method gave excellent recoveries of cocaine (98-100%) from spiked samples at concentrations of between 50 ng/ml and 10 micrograms/ml. Analysis of the extracts was by high performance liquid chromatography (HPLC).     

体液中微量士的宁的富集及同时检测

目的将分子印迹技术运用到法医毒物分析中,建立一种全新的测定体液中微量士的宁的方法。方法运用分子印迹原位聚合技术,在色谱柱中一步合成了对士的宁具有专属识别能力的分子印迹聚合物,以此聚合物为介质,装填一厘米的小柱,建立体液中微量士的宁的富集及同时检测方法。结果以士的宁分子印迹聚合物作为尿液和血浆中微量士的宁的富集和检测介质,方法的检出限为4.9ng,样品回收率均大于92%,相对标准偏差小于6.59%,工作曲线的线性相关系数分别为0.9991和0.9966。运用此方法对中毒家兔血浆和尿液中的士的宁进行了测定。结论建立的新方法可以实现对士的宁专属的、灵敏的检测,适合于法医学毒物分析运用。     

Different assays for hingamine in the study of biological fluids

The authors present a method of hingamine identification in non-biological substances (tablets, powder, syringes) and biological fluids (blood, urine). Isolation was made with chloroform in pH 10. Identification was conducted with thin-layer chromatography, gas chromatography/mass-spectrometry, high-performance liquid chromatography, spectrophotometry in UV region. The quantity was estimated with spectrophotometry in UV region, high-performance liquid chromatography and high-performance thin-layer chromatography. The results of the three methods are comparable.     

体液中常见滥用药物的系统筛选分析

本文建立了体液中常见滥用药物的筛选分析体系.尿液或血液经固相萃取(SPE)或液提取(LLE)后,直接用GC/NPD分析或经TFA、BSTFA衍生化后用GC/MS分析.方法适用于同时分析甲基苯丙胺、MDMA、度冷丁、去甲度冷丁、曲马多、美沙酮、EDDP、可卡因、苯甲酰芽子碱、可待因、安定、氯丙嗪、吗啡、单乙酰吗啡等十四种常见滥用药物及代谢物.SPE法和LLE法回收率分别为66～102%和50～86%,最低检出限为2-5ng/ml尿.涉毒案件的鉴定应用表明该分析方法简便、快速、可靠.     

生物试样中巴比妥类药物固相提取柱上衍生化GC/MS分析

建立生物试样中常见巴比妥类药物的固相提取和柱上衍生化GC／MS分析法。将预制的血或肝分别在pH6．0和pH2.2的条件下过预活化的GDX－403吸附小柱，再用缓冲浪和蒸馏水各4ml顺序洗柱。最后用4ml丙酮／氯仿（1：1）溶剂洗脱样品，离心弃除水相，80℃挥至近干，用50μl乙醇定溶、取净化的样品2～4μl挥至近干，加20μl0．2molTMAH衍生化试剂，直接进样0.5μl，柱上衍生化GC／MS（GC）分析。在试验条件下，当血和肝分别添加2．0μg和5．0μg混合药物，回收率≥80％，相对标准差（RSD）优于±10％，检测限优于5ng（信／噪比≥2）。该法能有效地排除类脂物和组胺的干扰，可用于治疗量级药物分析和婴幼儿中毒案检验。     

Acephate in biological fluids of two autopsy cases after ingestion of the chemical

Two autopsy cases, where the individuals were suspected of having ingested acephate, an organophosphorous insecticide, are reported. Acephate and its active metabolite, methamidophos (MP), were analyzed in the biological fluids by GC/MS, using the salting out method with liquid-liquid extraction columns. The first case was that of a 70-year-old man whose blood acephate was 149 microg/mL, and MP was 3.0 microg/mL. Serum pseudocholinesterase (ChE) activity was inhibited. No remarkable finding of injury or disease was determined as the cause of his death, but acute poisoning by acephate was mostly suspected. The second case was that of a 60-year-old man. A deep gash in the left neck injured the left common carotid artery in addition to the severely ischemic state of the primary organs. His blood acephate was 46 microg/mL, and MP was not detected. ChE activity was in the normal range. Hemorrhage was mainly suspected as the cause of his death. The concentrations of acephate and MP in human blood after oral ingestion are first reported here, and the acute toxic level of acephate is discussed.     

Automated extraction of DNA from reference samples from various types of biological materials on the Qiagen BioRobot EZ1 Workstation

We have validated and implemented a protocol for DNA extraction from various types of biological materials using a Qiagen BioRobot EZ1 Workstation. The sample materials included whole blood, blood from deceased, buccal cells on Omni swabs and FTA Cards, blood on FTA Cards and cotton swabs, and muscle biopsies. The DNA extraction was validated according to EN/ISO 17025 for the STR kits AmpF?STR^® Identifiler^® and AmpF?STR^® Yfiler^® (Applied Biosystems). Of 298 samples extracted, 11 (4%) did not yield acceptable results. In conclusion, we have demonstrated that extraction of DNA from various types of biological material can be performed quickly and without the use of hazardous chemicals, and that the DNA may be successfully STR typed according to the requirements of forensic genetic investigations accredited according to EN/ISO 17025.     

The chromatographic analysis of phencyclidine, its metabolites and analogs in biological fluids

Reviews data on analysis of narcotic phencyclidine and its main metabolites and analogs. Pharmacological and toxic characteristics of phencyclidine, conditions of its isolation from biological objects and its detection and measurement by thin layer and gas chromatography and by chromatographic mass-spectrometry are described. Analysis of phencyclidine metabolites is discussed and a scheme of its metabolism is presented. Statistical characteristics of methods of quantitative analysis of phencyclidine in biological objects are presented.     

The detection of tubazid and saliuzid metabolites in internal organs and biological fluids

Detection of tubaside and saluside metabolites by methanol extraction from homogenized object with subsequent investigation by UV spectrophotometry, microcrystalloscopy, thin-layer chromatography and coloured reaction methods is suggested. Sensitivity for hydrazine, acetylisoniazid was 0.1 mg%, for opianic and isonicotinic acids--0.05 mg%.     

Rapid extraction and capillary gas chromatography for diazine herbicides in human body fluids

A simple and rapid method for the extraction of four diazine herbicides (terbacil, bromacil, norflurazon and PAC) from human whole blood, plasma and urine with use of Bond Elut C₁₈ cartridges is presented. Whole blood, plasma and urine samples containing the herbicides, after mixing with distilled water, were loaded on Bond Elut C₁₈ cartridges and the herbicides were eluted with chloroform/methanol (9:1). They were detected by capillary gas chromatography with flame ionization detection (FID) with splitless injection. Separation of the four diazine herbicides from each other and from impurities was generally satisfactory with the use of an intermediately polar DB-17 capillary column. The recovery of all compounds, which had been added to whole blood, plasma and urine, was > 89%. The calibration curve for the herbicides, which has been added to whole blood, plasma and urine, showed linearity in the range 1.6–100 ng on column. Their detection limits were 1.2–1.4 ng on column for whole blood and plasma, and 1.1–1.2 ng on column for urine.     

Determination of glyphosate in biological fluids by 1H and 31P NMR spectroscopy

Identification of glyphosate in four cases of poisoning, using nuclear magnetic resonance spectroscopy of biological fluids is reported. It has been performed by using a combination of 1H and 31P NMR analyses. Characterization of the N-(phosphonomethyl) glycine herbicide was achieved by chemical shift considerations and coupling constant patterns: CH2-(P) presents specific resonance at 3.12 ppm and appears as a doublet with a H-P characteristic coupling constant of 12.3 Hz. Moreover, resonances due to isopropylamine were present, confirming the ingestion of the considered trade formulation. After a calibration step, quantitation was performed by 1H and 31P NMR spectroscopy. The benefit and reliability of NMR investigations of biological fluids are discussed, particularly when the clinical picture is quite confusing.