| The non-Benzodiazepines drugs(NBZD) is an inhibitor which has direct role in the central nervous system, belongs to the third generation of sedative hypnotics. The drug appeared in the late 1980 s, azole pyrazole(Zolpidem), Zopiclone(Zopiclone),(Zaleplon) as the representative drugs, which have a function of sedative hypnotic, anti-anxiety, anticonvulsants and muscle relaxation, mainly used in the treatment of insomnia. This class of drugs(BZ) has a specific selectivity to benzodiazepine receptors, and fall asleep quickly, significantly increase the deep sleep, while don’t change the basic physiological structure of normal sleep, wake up without hangover feeling.Because of its consumption, good hypnotic effect, quick effect, so it is effective to meet insomnia people at the same time, some criminals also use the new type hypnotics commit suicide, anesthesia robbery, murder, anesthesia rape or other crimes, therefore, the suicides in society, the crime occurred frequently. So it is necessary to establish a rapid and effective identification method, to match the clinical rescue work and the court of criminal case detection. And to provide scientific evidences to determine whether the subject has taken such drugs and what drugs the subject has taken.Ultra high performance liquid chromatography- mass spectrometry(UPLC/MS/MS) has efficient liquid phase separation ability and high sensitivity and strong structure of mass spectrum, and it is a specific, sensitive, rapid accurate and reliable to become a powerful analysis tool for chemical identification, this study adopted the UPLC –MS/MS technology to establish the non-benzodiazepines sedative hypnotics analysis method, the zopiclone, Eszopiclone, Zaleplon in plasma and urine samples were measured, and provides a simple, rapid and reliable method for the rapid identification, quantitative analysis of this class of drug. The first part Detection of dexzopiclone, zolpidam and zaleplon in the human plasma by UPLC/MS/MS methodObjective:To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) for determination of dexzopiclone, zolpidam and zaleplon in human plasma.Methods:The plasma samples were extracted with dichloromethane:n-hexane:acetoacetate(5:4:1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column(1.8 μm,2.1×100 mm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 ml/min in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring(MRM) via electrospray ionization(ESI) source in positive mode.Results:Good linear relation were obtained over the investigated concentration range, with all correlation coefficients higher than 0.99. LOQ, precision, accuracy, stability, extraction and recovery of the matrix effect meets the requirements. Extraction recovery range of 87.4%~108.0%, RSD was 4.5%~14.6%.Conclusion:It is simple, rapid, high sensitivie, and can be used to determination of Zaleplon, zopiclone and dexzopiclone in human plasma at the same time. The second part Detection of dexzopiclone, zolpidam and zaleplon in the human urine by UPLC/MS/MS methodObjective: To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) for determination of dexzopiclone, zolpidam and zaleplon in human urine.Methods: The urine samples were extracted with dichloromethane:n-hexane:acetoacetate(5:4:1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column(1.8 μm,2.1×100 mm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 ml/min in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring(MRM) via electrospray ionization(ESI) source in positive mode.Results:Good linear relation were obtained over the investigated concentration range, with all correlation coefficients higher than 0.99. The precision of three analytes(RSD) are less than 15.0%. Extraction recovery were range of 79.8 ~ 105.5%, the matrix effect were range of 0.85 ~ 1.09. After three freezing- thawing cycles, the difference of initial concentration between quality control samples and fresh sample is within 15.1%, proved that stability is good after three freezing- thawing cycle. LOD, precision, accuracy, stability, extraction and recovery of the matrix effect meets the requirements.Conclusion:It is simple, rapid, high sensitivie, and can be used to determination of Zaleplon, zopiclone and dexzopiclone in human urine. |
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