A single-column procedure on Bond Elut Certify for systematic toxicological analysis of drugs in plasma and urine.

A single-column solid-phase extraction procedure was developed for the screening of acidic, neutral, and basic drugs from plasma. The recoveries of all 25 tested drugs exceeded 82%. After the plasma had been diluted with phosphate buffer (pH 6.0), the drugs were extracted using a single Bond Elut Certify column. The acidic and most of the neutral drugs were eluted by acetone/chloroform (1:1) and the basic drugs were eluted by 2% ammoniated ethyl acetate. Some neutral drugs appeared in both fractions. The two fractions were collected separately and evaporated until approximately 100 microL of solvent remained in the tube. Both fractions were analyzed separately on a gas chromatograph equipped with a wide-bore capillary column and a flame ionization detector. The procedure could also be used for urine samples.     

腐败生物检材中多种碱性滥用药物的检测

目的建立腐败生物检材中多种碱性滥用药物的提取、净化和仪器分析方法。方法用环己烷作为提取溶剂液-液萃取,同时采用Bond Elut Certify小柱、甲醇淋洗、二氯甲烷:异丙醇:氨水(78:20:2)洗脱固相萃取分离提取,GC/MS、GC/NPD定性定量分析各种生物检材中的滥用药物。结果从所送死者肝组织、胃组织、心血及胃内容、尿样、各检材中均同时检出吗啡、可待因、舒乐安定和异丙嗪成份,其中肝组织含量分别为吗啡0.094μg/g、可待因0.257μg/g、异丙嗪0.110μg/g,尿液含量分别为吗啡0.334μg/ml、可待因4.054μg/ml、异丙嗪0.066μg/ml,心血含量分别为吗啡0.036μg/ml、可待因0.106μg/g、异丙嗪0.088μg/ml。结论此方法准确、可靠、科学,可以用于法医毒物分析领域体内检材多种碱性药物的检测。     

Isolation of drugs from blood and tissues with XAD-2 bags

Nylon bags containing 2-g portions of Amberlite XAD-2 resin were used for systematic analysis of drugs in biosamples. The procedure requires 10 or less grams of material, two XAD-2 bags, and enables rapid and economical isolation of most common drugs. The method was demonstrated on autopsy blood spiked with 19 of the most common drugs, and was routinely used in cases of fatal and non-fatal poisoning. The eluates were clean and suitable for direct gas chromatographic and ultraviolet spectrophotometric analysis.The procedure used appeared more convenient than XAD-2 column extraction procedures. Classic solvent extraction methods were usually less efficient.     

UPLC-MS/MS法检测血痕中常见烟草、毒品和药物成分

目的建立UPLC-MS/MS测定血痕中尼古丁、可替宁、甲基苯丙胺、氯胺酮、吗啡、O6-单乙酰吗啡、地西泮、三唑仑、艾司唑仑、佐匹克隆和利血平的方法。方法以甲醇为提取液,采用基质提取溶液配制标准溶液制作定量曲线。采用超高效液相色谱柱对待测样品进行分离;以电喷雾离子源正离子（ESI＋）模式和多反应监测（MRM）模式进行质谱分析。优化实验条件,并进行方法学评价。结果选择UPLC HSS T3色谱柱,乙腈-5mmol/L甲酸铵＋0.1%（v/v）甲酸作为流动相,甲醇作为提取溶剂。11种检测物检出限（S/N=3）和定量限（S/N=10）分别为0.04~2.82ng和0.13~7.64ng,1~500ng/mL范围内的线性关系良好。除利血平、吗啡外的检测物回收率为（53.8±5.3）%~（107.2±12.6）%。结论采用本文UPLC-MS/MS方法对血痕中11种常见烟草、毒品和药物成分进行检测,能够满足实际检案的要求,可在相关检验中选用。     

固根萃取技术在系统药、毒物分析中的应用

根据反相色谱法及离子交换色谱法的原理,研制了一种固相萃取小柱,建立了筛分系统,可以同时完成对复杂体系中酸性、中性及碱性药、毒物的一次革取净化及富集,适应于系统毒物及药物筛分工作的需要。并对筛分柱的草取机理进行了初步的实验论证。     

Extraction of selected drugs from serum using microwave irradiation

Microwave irradiation is used as an alternative heating method for extraction over more conventional hot plate methods. We describe a fast, efficient method for the determination of selected drugs in human blood/serum using microwave extraction. The microwave extraction of organic substances requires special instrumentation and the results have been compared with the results from classical liquid/liquid extraction. The present microwave extractions were performed in an ‘atmospheric pressure’ system. Before irradiation with microwaves, an appropriate solvent mixture was added to the buffered specimen. Lidocaine, methadone, diazepam, nordiazepam, propoxyphene and norpropoxyphene were tested as model substances. The quantitation was performed by GC/NPD. The procedure has been applied successfully to a number of forensic cases. The use of microwaves decreases the time of extraction and the solvent consumption.     

Detection of drugs using XAD-2 resin. I:Choice of resin, chromatographic conditions, and recovery studies

Amberlite XAD-2, a nonionic polystyrene divinylbenzene resin, was first used for the analysis of drugs in urine and a number of reports have described the development at optimal conditions for extraction, including type of resin columns, pH conditions, and eluting solvents. XAD-4 and XAD-7 resins were compared to the similarly structured XAD-2 resin and no significant advantage over the XAD-2 resin for drug screening was observed. A quantity of 5 to 6 g of resin was found to have sufficient capacity for the extraction of 200 ml of pentobarbital solution (1 mg/100ml). A column flow rate of approximately 15 ml/min (gravitational flow) was sufficient for analysis and slower rates were not more efficient. A mixture of ethyl acetate and 1,2-dichloroethane (3:2) was found to give best overall recovery (66 to 94%) of drugs, the resulting extracts being reasonably free of interfering substances. A pH value of 8.5 is recommended as optimum for comprehensive analysis of acidic and basic drugs. Recovery studies were conducted on spiked samples to determine drug losses occuring during various steps in the XAD-2 extraction procedure for four acidic (amobarbital, secobarbital, pentobarbital, and phenobarbital) and four basic (morphine, codeine, meperidine, and methadone) drugs. A relatively small amount (0 to 5%) of the drugs was not adsorbed by the resin and amounts varying from 6 to 40% failed to be desorbed by the eluting solvent. Additional losses occurred during the removal and analysis of TLC spots. Recovery of drugs from aqueous solutions analyzed with the XAD-2 resin were compared to recoveries reported in the literature with other XAD-2 resin methods for the extraction of drugs from urine. Recovery of phenobarbital, morphine, and codeine improved by 4 to 23% while recoveries of amobarbital, pentobarbital, secobarbital, methadone, and meperidine were 4 to 28% less efficient when compared to literature data.     

Rapid extraction of oxazepam from greyhound urine for high performance liquid chromatography analysis

A rapid and efficient procedure is described for the extraction and analysis of oxazepam, the major urinary metabolite of diazepam in greyhounds. Urine was extracted by passing through a bonded silica column (Bond Elut) following enzyme hydrolysis. The adsorbed drug was eluted and then detected and measured by high performance liquid chromatography (HPLC). Recoveries were in excess of 85% at 50 ng/ml concentrations. Detection was possible up to 30 h after a single oral dose of diazepam (5 mg).     

The differential elution of drugs from XAD-2 resin.

Biological fluids and tissue extracts prepared according to a previously published method were passed through a column of Amberlite XAD-2 resin for removal of drugs. The differential elution of the adsorbed drugs from the resin was performed by sequential elution of the drugs in four steps. The column was first washed with 30 ml of 0.05M sodium acetate buffer, pH 4.55. Drugs exhibiting acidic or neutral characteristics were then eluted from the column with 100 ml of chloroform in 20-ml aliquots. The column was then washed with 30 ml of 0.1M potassium carbonate, which was discarded. Drugs exhibiting basic characteristics were then eluted from the column with 100 ml of chloroform:isopropanol (3:1) in 20-ml aliquots. Sensitivity of drug detection with this method by thin-layer chromatography was 0.5 mug/ml in a 10-ml sample for nearly all drugs tested.     

Simultaneous screening and quantitation of 18 antihistamine drugs in blood by liquid chromatography ionspray tandem mass spectrometry

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is presented for the simultaneous screening and quantitation of 18 antihistamine drugs in blood samples. Sample pretreatment involved liquid-liquid extraction of the basic antihistamines followed by a second extraction of the acidic antihistamines. The recoveries were 43-113% for basic drugs and 23-66% for acidic drugs. The combined extracts were run by LC on C(18) reversed phase column using acetonitrile-ammonium acetate mobile phase at pH 3.2. The mass spectrometric analysis was performed with a triple stage quadrupole mass analyzer. Screening was performed using multiple reaction monitoring (MRM) and any compounds tentatively identified as antihistamine drugs were then automatedly verified by their Product Ion Spectra in a subsequent MS/MS run.Quantitation was based on the MRM data from the screening step. In validation tests, the method showed good linearity at the relevant concentrations. The attained limits of quantitation varied between 0.0005 and 0.01mg/l in blood and were lower than the therapeutic concentrations (C(max)). The limits for identification by Product Ion Spectra were also lower than C(max), except for clemastine, which has exceptionally low concentrations in blood. The intra-assay relative standard deviations were better than 10% and the inaccuracy varied between 39% for levocabastine and 5% for cyclizine, the majority of the values being <20%.     

快速溶剂萃取-气相色谱/质谱法分析血液中的4种滥用药物

目的 建立生物检材血液中滥用药物的快速溶剂萃取（ASE）方法.方法 通过优化快速溶剂萃取各参数,提取血液中的滥用药物进行GC/MS定性定量分析.结果 血液中美沙酮、可卡因、蒂巴因、海洛因4种滥用药物的平均回收率在87.8％～97.5％之间,滥用药物在0.4μg/mL～4.0μg/mL的浓度范围内线性良好.结论 该方法具有操作简便快捷,回收率高、重现性好等特点,可广泛应用于生物检材血液中滥用药物的检验鉴定.     

An evaluation of fused silica capillary columns for the screening of basic drugs in postmortem blood: qualitative and quantitative analysis

Fused silica capillary columns (Durabond) have been evaluated for the screening of more than 100 basic drugs in postmortem blood samples. The combination of these columns, nitrogen-phosphorus detectors, and SKF-525A (internal standard) allows for the simultaneous screening and quantitation of several basic drugs such as amphetamines, amitriptyline, and codeine. Approximately 2000 blood samples have been analyzed by this procedure. The use of capillary columns results in excellent baseline stability and this, together with an autosampler and data system, enables unattended overnight operation. "Double peaking" associated with splitless injection can be a problem as can sensitivity for some of the polar drugs; however, with the extraction procedure described and the equipment used, the screening of blood for basic drugs is improved when compared with packed column technology.     

Evaluation of a photodiode array/HPLC-based system for the detection and quantitation of basic drugs in postmortem blood.

A systematic analytical approach has been developed for liquid chromatography determination of a number of basic drugs in postmortem blood. Using selective extraction, that is, back extraction into 0.2N sulfuric acid and 6N hydrochloric acid after the initial extraction with toluene under basic conditions (from 2 mL of blood), basic and weakly basic drugs, such as propranolol and diazepam, can be simultaneously quantitated and identified with a high degree of confidence. A microcomputer-based photodiode array detector was used to evaluate peak purity and facilitate peak identification. An automatic library search was performed at the end of each analysis using the system software. The method was validated for within-day and between-day precision for ten basic drugs at two concentrations. The coefficient of variation for the between-day precision was less than 8.7%. Accuracy of the assay was tested at four concentrations using linear regression analysis. The coefficients of determination (r2) for all ten drugs were greater than 0.99, and their slopes were close to unity. The chromatographic conditions developed are suitable for the screening of several basic, acidic, amphoteric, and neutral drugs. Retention data and ultraviolet spectral data for 119 drugs on two reversed-phase columns, using acidic mobile phases, are also presented.     

Application of solvent microextraction to the analysis of amphetamines and phencyclidine in urine

A fast and simple method to detect some commonly abused illicit drugs, amphetamine, methamphetamine, 3,4-methylendioxy-amphetamine (MDA), 3,4-methylendioxy-methamphetamine (MDMA), 3,4-methylendioxy-N-ethylamphetamine (MDEA) and phencyclidine (PCP) in urine using solvent microextraction (SME) combined with gas chromatography (GC) analysis has been developed. The extraction is conducted by suspending a 2 microl drop of chloroform in a 2 ml urine sample. Following 8 min of extraction, the organic solvent is withdrawn into the syringe and injected into a GC with a pulsed discharge helium ionization detector (PDHID). The effects of different extraction solvents and times, pH and sample preparation were studied. The optimized method was capable of detecting drugs in urine at concentrations below Substance Abuse and Mental Health Services Administration (SAMHSA) established cut-off values for preliminary testing. Good linearity and reproducibility of extraction were obtained. The limits of detection were 0.5 microg/ml for amphetamine, 0.1 microg/ml for methamphetamine and MDA, 0.05 microg/ml for MDMA, 0.025 microg/ml for MDEA and 0.015 microg/ml for PCP. Relative standard deviation (R.S.D.) values ranged between 5 and 20% for the studied drugs.     

Simultaneous identification and quantification of several opiates and derivatives by capillary gas chromatography and nitrogen selective detection

A capillary column gas chromatographic method is described for the simultaneous determination of morphine, codeine, heroin, 3- and 6-monoacetylmorphine, nalorphine, naloxone, ethylmorphine, and naltrexone. The drugs were extracted from 2 ml plasma, urine, or other biological samples, including tissue under alkaline conditions in chloroform-isopropanol-n-heptane (50:17:33, v/v), with levallorphan as an internal standard. The drugs were extracted into acid and then reextracted into chloroform after the acid had been alkalinized. After derivatization with trifluoroacetic anhydride, an aliquot was injected into a 25m capillary column equipped with a nitrogen phosphorus detector. The lower limits of detectability, extraction recovery, and the within-run and day-to-day precision of results were determined for each drug. Our results indicate that the procedure is suitable for use in overdose screening and therapeutic drug monitoring.     

快速溶剂萃取法及其在刑事技术中的应用

快速溶剂萃取技术是近几年出现的一种新的适用于复杂检材前处理的、高效、快速、简便、理想的萃取方法。快速溶剂萃取法的原理、特点、仪器装置与操作步骤、影响因素在刑事技术领域的应用。     

Comparison of extraction methods for methamphetamine and its metabolites in tissue

Five extraction methods were examined for analysis of methamphetamine and its major metabolites in tissue samples. The extraction methods studied were an acetone extraction method, an ethanol extraction method, an ammonium sulfate method, dialysis, and a direct solvent extraction. Acetone, ethanol, and dialysis methods showed no interference from endogenous components using thin-layer chromatography and gas chromatography, and gave satisfactory recovery of methamphetamine, amphetamine, and p-hydroxymethamphetamine when added to rabbit liver. These methods, however, proved time-consuming. The ammonium sulfate method and direct solvent extraction method were simple and more rapid, but recovery of the polar metabolite was poor.     

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.     

Solid-phase extraction and analysis of 20 antidepressant drugs in human plasma by LC/MS with SSI method

A simultaneous determination of 20 antidepressant drugs (imipramine, amitriptyline, desipramine, trimipramine, nortriptyline, clomipramine, amoxapine, lofepramine, dosulepin, maprotiline, mianserin, setiptiline, trazodone, fluvoxamine, paroxetine, milnacipran, sulpiride, tandspirone, methylphenidate and melitracen) in human plasma was developed using LC/MS with sonic spray ionization (SSI) method. These drugs showed good separation and sensitivity by LC-MS using an Inertsil C-8 column with methanol:10mM ammonium acetate (pH 5.0):acetonitrile (70:20:10) as mobile phase at 0.10 mL/min at 35 degrees C. Solid-phase extraction of these drugs added to the human plasma was performed with an Oasis HLB cartridge column. Recovery and limit of detection of these drugs were between 69 and 102% and between 0.03 and 0.63 microg/mL, respectively. The present procedure offers an easier and more convenient screening method for antidepressants, and will be useful for forensic toxicology investigations.     

A routine screening method for the major metabolite of methyl phenidate in the urine of greyhounds

A qualitative urine screening procedure for the determination of the major urinary metabolite of methyl phenidate, α-phenyl-2-piperidine acetic acid (ritalinic acid), has been devised.The method involves a salting-out extraction, conversion of the metabolite to the parent drug using methanol-hydrogen chloride, and simultaneous two-column gaschromatographic detection. Final confirmation of the presence of the drug is by two-system thin-layer chromatography. The method is suitable for the detection of illicitly administered methyl phenidate in greyhounds.