Screening Of Drugs Of Abuse,Pharmaceuticals And Their Metabolites In Blood And Urine By HPLC-LTQ-Orbitrap-MS

Due to the diversity of toxicologyically relevant substances, the uncertainty of untarget compounds and the specificity of samples, toxicological screening techniques have always been valued by the forensic toxicologists. It is often necessary for forensic toxicology scientistis to perform a systematic screening method for any toxicologically relevant substances. Therefore, toxicological screening has always been an important part in forensic toxicology and clinical toxicology. In our laboratory, the target analytes includes drug of abuse, pharmaceuticals, pesticides, rat poison, anabolic hormones and etc.Liquid chromatography-high resolution mass spectrometry (LC-HRMS) is a new technique which combines liquid chromatography with high-resolution mass spectrometry. In recent years, the high-resolution mass spectrometer was developped from large-scale equipment to desktop instrument, and it has been commonly used in laboratories. Due to the high resolution, accurate mass can be obtained by HRMS, which made it possible to distinguish co-eluting impurities from complex matrix. In theory, more than a hundred compounds can be simutenouly determined by LC-HRMS. Furthermore, owing to the full-scan acquisition at good sensitivity, LTQ-Orbitrap-MS offers the possibility to investigate the presence of compounds once the analysis has been performed and data required, without being dependent on the pre-selection of analytes. It is a target and non-target screening methods, which has been applied in forensic science.A rapid screening method was developped for1000drugs of abuse, pharaceuticals, pesticides, hormones. The method is based on an accurate mass database. The screening criteria were studied and discussed. HPLC-LTQ-Orbitrap methods for500compounds were developed and validation of the method was performed.1. Screening method for1000common toxicological relaventsDevelopped a screening method for1000common toxicological relavants. The method is based on an accurate mass database, which includes the compound formula, retention time (if available) and other information. A HPLC-LTQ-Orbitrap MS method was developped for500most common drugs of abuse (including illicit drugs, opiates, amphetamines, cannabinol), pesticides, organophosphorus, carbamate, benzodiazepine, epilepsy, asthma, antipyretic analgesics, antibiotics, and other common therapeutic agents, etc. Established two liquid chromatography separation system, the liquid pentafluorophenyl (PFP) column was used to analyze phenyl-containing alkaline compounds, the C18column was used to analyze polarity (neutral and weakly acidic) compounds. Using electrospray ionization source (ESI) in both positive mode and negative mode with full scan, the resolution was set at FWHM60,000for full scan analysis.2Screening and determination of drugs of abuse in urine by HPLC-LTQ-by OrbitrapA HPLC-LTQ-Orbitrap-MS screening method for63drugs of abuse was developed. With morphine-D3, methamphetamine-the D8, cocaine-D3as internal standard, the target compounds in the urine sample were enriched and purified by Oasis HLB solid phase extraction column, then separated by Hypersil Gold PFP column with0.1%formic acid water and acetonitrile, finally analyzed by LTQ-Orbitrap MS using ESI+full scan mode, resolution of FWHM60,000. The limit of detection of most compounds is less than lng/mL. Recovery was>50%and matrix effect was>70%, respectively. RSD were less than15%. The developed method is high sensitivity, good selectivity, and is appropriate for the analysis of drugs of abuse in forensic toxicology.3. Screening and confirmation of psychotropic drugs in blood and urine by HPLC-LTQ-Orbitrap MSDevelopped a screening and confirmation method for psychotropic drugs and their metabolites in human blood and urine by HPLC-LTQ-Orbitrap MS. The samples were pretreated with SiroccoTM protein precipitation plate, then separated by Hypersil Gold PFP column with0.1formic acid water and acetonitrile, finally analyzed by HPLC-LTQ Orbitrap MS, using ESI+full scan mode with a resolution of FWHM60,000. Collision-induced dissociation (CID) was employed for the qualitative accurate mass of fragmen ions.The method was validated in terms of the limit of detection. LOD of psychotropic substances was<0.0lng/mL, except lorazepam5ng/ml, N-methyl flunitrazepam20ng/ml, Mass accuracy for all compounds were less than1ppm. An accurate mass was created for psychotropic drugs screening. The developed method is simple, fast, accurate, and meet the need of court toxicology analysis. The method established by HPLC-LTQ-Orbitrap-MS effectively solved the problems that exist in the current study of drugs of abuse.In summary, the innovation of this paper are as follows:1The accurate mass database contains more than1,000toxicological relavants, covering drugs of abuse, theraputical drugs and their metabolites. Two liquid chromatography separation systems were established to ensure that most of the compounds showed a good chromatographic behavior in a liquid chromatography system. The retention time of500kinds of toxicological relevants were obtained. The developped screening method can also be used for compounds determination. The study meets the needs of the proof in forensic sciences, coming to the advanced international level.2The HPLC-LTQ-Orbitrap MS method for analysis of morphine, cocaine, amphetamines, hallucinogens, psychotropic in biological samples were developed, and the validation of the method was perfomed in biological matrix. The methods are for the first time in our country.