| Summary: | 博士 === 中山醫學大學 === 醫學分子毒理學研究所 === 101 === Due to the drug abuse with items increasing and complicate, traditional urine test could easily lead to the loopholes of drug testing, and therefore need to develop alternative biological specimen testing methods to enhance the deterrent capability of drug test. However, drug concentration in alternative biological specimens was lower than in the urine, thus the method developments of reduced-size sample analysis and trace concentration drug detection were limited.
The purpose of this thesis is to develop sensitive hair and oral fluid testing method for the determination drugs of abuse. The 13 international common drug and its metabolites were selected, including amphetamine (AP), methylamphetamine (MA), methylenedioxy amphetamine (MDA), methylenedioxy methmphetamine (MDMA), Tetrahydrocannabinol (THC), tetrahydro cannabinolcarboxylic acid (THCA), morphine (MOR ), 6-acetyl morphine (6-AM), codeine (COD), cocaine (COC), benzoylecgonine (BZE), that ketamine (K), norketamine (NK).
In order to elevate the sensitivities on liquid chromatography - mass spectrometry (LC-MS), the strategy of pre-column chemical derivatization with dansyl chloride was employed in this thesis to modify the properties of analytes. The results showed: after dansyl chloride derivatization, the AP, MA, MDA, MDMA, THC, THCA, MOR, 6-AM could be derivated successfully. In addition, the sensitivity on phenolic compounds could be enhanced significantly on positive APCI.
Subsequent study further applied to simultaneous analysis of multi-drug abuse in reduced-size hair sample on ESI. The method was validated and applied to the real reduced-size sample hair (1 mg). Compared to the developed method on gas chromatography - mass spectrometry (GC-MS), the quantification results showed high consistence on LC-MS after dansyl chloride derivatization.
On oral fluid testing, due to the low concentration of THC-COOH in oral fluid, caused there is no LC-MS based method was developed with sufficient sensitivity. Thus, this LC-MS after derivatization method was applied to the real cannabis abusers oral fluid sample detection. Six suspect real samples could be further detected the THC-COOH when THC was positive (> 2 ng/mL). This is our current knowledge on the most sensitive LC-MS method.
On the other hand, due to the complication of preparation process on hair test, caused the time waste. Thus, desorption electrospray ionization (DESI) was applied to direct and fast detection abused drug in hair. We modified the laboratory LCQ classic ion trap mass spectrometer, and to install a straight line moving loading platform. The interface was called scanning-DESI. The parameter on spray solvent flow rate, spray voltage, and sample plate moving rate of scanning-DESI was optimized with rhodamine B. Then the sensitivity to drug abuse standards was evaluated. The result showed the lowest detection limit for AP, MDA and NK were 5 pg/mm2, MA, MDMA and K were 10 pg/mm2; for 6-AM was 20 pg/mm2; for morphine was 50 pg/mm2. The interface was applied to direct and fast detection on real high concentration hair sample, and further used on hair drug footprint scanning, and in-depth analysis of the drug distribution.
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