Determination of urinary markes of PAH and toluene exposure by dispersive liquid-liquid microextraction based on solidification of floating organic droplet/liquid chromatography- fluorescence

• Main Authors: Yi-Lin Chen, 陳奕霖
• Other Authors: 沈振峯
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/cncuc7

碩士 === 國立虎尾科技大學 === 生物科技系碩士班 === 104 === Toluene and polycyclic aromatic hydrocarbons(PAHs) were common hazard materials on the industry, long-term exposure to toluene and PAHs can cause central nervous system damage and cancer respectively.In this stady, simultaneously the biological detection method for hazard, shorten the sample preparation time and reduce the cost of analysis. Urinary 1-hydroxypyrene(1-OHP) and o-cresol are main metabolites of the two types of hazards, and detected with liquid chromatography-fluorescence(LC-FL).The technique of dispersive liquid-liquid microextraction based on solidification of floating organic droplet method (DLLME-SFO)is used for sample processing.At first 3 mL urine was hydrolyzed for 2 hours by enzyme of β-glucuronidase.followed by addition of 50 μL 1-undecanol(extraction solvent) and 1 mL acetone(dipsersive solvent) for DLLME-SFO(1 min).After centrifuged and frozen 30 min in-20 ℃, the solidification droplet(ice) was mixed by equal volumes methanol, transferred into 1.2 mL vial and injected to LC-FL analysis.The blank urine was stripped by charcoal powder and compared the relative standrd deviation with other source urine, the result of relative standrd deviation was less than 20 %. The developed method was calidated with spiked blank urine samples. The calibration curve was calculated with area against the spiked concentration. The linear ranges of the urinary markers were o-cresol: 2.81-180.00 ng mL-1、1-OHP:37.50-300.00 pg mL-1, the satisfid linearity was obtained for each markers (r2>0.99) , the obtained detection limits(S/N ratio≧3) were o-cresol: 0.35 ng mL-1, 1-OHP: 9.38 pg mL-1, the accuracy ( % Error) and precision(CV %) were range 3.74-12.22 %and ±20 %, respectively(3 spiked levels, 6replicates). The method was applied for 7 clinical smaples, the presence of o-cresol and 1-OHP were detected.