DPHP 和DINCH為DEHP與DINP的替代性塑化劑。DPHP 及DINCH使用量逐漸增加，這也提高了一般民眾暴露的可能性，而利用量測尿液中DPHP及DINCH的代謝物訊號可以評估DPHP及DINCH暴露的情形。因此本研究目的為利用高解析質譜儀及實驗室所開發的同位素標定追蹤方法(SMAIT)找尋體外培養樣本中DPHP及DINCH代謝物訊號，並用動物實驗驗證為暴露指標。樣本經由人類肝臟酵素進行24小時反應，以固相萃取裝置進行樣品淨化，接著以液相層析質譜儀進行樣品分析，分析時分別以負離子及正離子模式掃描，產生的質譜數據以SMAIT進行訊號篩選，在負離子模式分析中篩選出12個DPHP與18個DINCH可能的代謝物訊號，在正離子模式中篩選出5個DPHP與8個DINCH可能的代謝物訊號。接著，利用動物實驗進行代謝物之驗證，將老鼠分為7組，每組3隻，分別暴露7種不同劑量之DPHP與DINCH(0、75、150、300、600、1200、2400 mg/kg body weight)，收取暴露後24小時尿液，以固相萃取進行尿液樣本淨化，接著以高解析質譜儀進行分析。動物實驗之驗證結果，15個DPHP (負離子模式11個、正離子模式4個)與16個DINCH (負離子模式9個、正離子模式7個)代謝物訊號可以在老鼠尿液中被量測到，且代謝物訊號強度與暴露劑量呈正相關(r ≥ 0.7, p ≤ 0.001)。將SMAIT與其他找尋代謝物之方法進行比較，雖然SMAIT篩選得到之代謝物訊號數量較少，但是經過動物實驗驗證結果，SMAIT在篩選代謝物的效益較好。結果顯示，經由SMAIT篩選並經過動物實驗所驗證所得到的15個DPHP及16個DINCH代謝物，可作為DPHP與DINCH暴露評估的指標。

Di-2(propylheptyl) phthalate (DPHP) and di(isononyl) cyclohexane-1,2-dicarboxylate (DINCH) are substitutes for some phthalates like di-(2-ethylhexyl) phthalate (DEHP) and di-iso-nonyl-phthalate (DINP). The increase of DPHP and DINCH consumption suggests a possible exposure of the general population. Exposure of DPHP and DINCH can be estimated by measuring concentration of metabolites. The aim of this study is to identify biological markers of DPHP and DINCH exposure by using high resolution mass spectrometry (HRMS) and signal mining algorithm with isotope tracing (SMAIT) in the in vitro samples, and validate the exposure-related markers in a rat model. In vitro DPHP and DINCH incubation samples wew incubated with human liver enzyme (S9 fraction) for 24 hours, and solid phase extraction (SPE) was used to clean up and condense those samples. Those samples were analyzed by HRMS, and the SMAIT program was used to identify metabolite marker candidates from the HRMS data. The 12 DPHP and 18 DINCH metabolite marker candidates in negative ion mode and 5 DPHP and 8 DINCH candidates in positive ion mode were identified by using SMAIT, and validated by rat model. The rats were divided into seven groups (N=3), and the 24-hour urine were collected after the rats orally administerated with different dose (0, 75, 150, 300, 600, 1200, 2400 mg/kg body weight). The results of rat model showed that the intensity of 15 DPHP (11 in negative and 5 in positive) and 16 DINCH (9 in negative and 8 in positive) metabolite candidates were positively correlated with increasing administrated dose (r ≥ 0.7, p ≤ 0.001). The more metabolite candidates can be identified by Mass Defect Filter (324 DPHP and 299 DINCH) than SMAIT. Nevertheless, the efficiency of identify exposure markers in SMAIT (92% for DPHP and 50% for DINCH) was better than MDF (7% for DPHP and DINCH). The result suggested that the 15 DPHP and 16 DINCH metabolite signals can be used as exposure markers of DPHP and DINCH.

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