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標題: | 檢測人體唾液鈦、鉻、砷、鎘、鉛之最佳化分析方法探討 Optimization of an Analytical Method for the Determination of Titanium, Chromium, Arsenic, Cadmium and Lead in Saliva |
作者: | 卓芸郁 Yun-Yu Cho |
指導教授: | 黃耀輝 Yaw-Huei Huang |
關鍵字: | 鈦,鉻,砷,鎘,鉛,唾液,感應耦合電漿質譜儀, titanium,chromium,arsenic,cadmium,lead,saliva,inductively coupled plasma mass spectrometry, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 生物偵測可反映個人在採樣當下的身體內金屬暴露負擔。而唾液樣本採樣簡單、便宜且侵入性低,具有作為生物偵測工具的潛在價值,但採樣方法、樣品前處理和分析儀器皆可能影響唾液樣本的分析結果。先前研究使用的唾液分析方法並不一致,且較少探討使用唾液樣本進行人體內金屬暴露生物偵測的效能。
本研究的目的分為兩部分。第一,開發並驗證一種使用感應耦合電漿質譜儀(inductively coupled plasma mass spectrometry)檢測人體唾液中鈦、鉻、砷、鎘和鉛的方法,並評估不同的樣本前處理方式對金屬濃度分析結果的影響,以優化分析方法。第二,招募20名健康成年志願者為受試者,並使用上述新開發的方法來測定人類唾液中的金屬濃度,同時利用問卷調查受測者的人口學、生活習慣、飲食頻率及口腔衛生相關的資料。 分析唾液樣本金屬濃度的前處理方法,首先使用人工唾液作為檢測唾液中金屬濃度的檢量線基質。唾液樣本以3500 rpm(≅1000 xg)離心10分鐘,取上清液後保存於4℃冰箱,並於5天內分析完。分析前,每個唾液上清液樣本以1%硝酸稀釋5倍,充分混合及過濾後進行金屬測定。該方法在測定鈦、鉻、砷、鎘和鉛的準確度分別為84.0%、102.7%、107.8%、116.3%、117.8%;且重複性及再現性皆小於8%。根據上述新開發方法所測定的受測者唾液中鈦、鉻、砷、鎘和鉛的平均濃度分別為140.2 ± 34.1 µg/L、0.21 ± 0.14 µg/L、0.16 ± 0.06 µg/L、0.03 ± 0.03 µg/L、0.85 ± 1.14 µg/L。此外,在兩小時內採樣五次的樣本分析結果發現,唾液中金屬平均濃度非常相近,顯示不同時點採樣的唾液中金屬濃度穩定。 本研究開發的方法僅需要簡單的樣品前處理,可準確地測量唾液中金屬濃度,並期許可作為人體暴露於金屬後之健康風險評估的重要工具之一。 Biomonitoring could reflect individual's current body burden of metal exposure. Saliva sampling is simple, inexpensive and non-invasive, leading it a valuable medium for the assessment of human exposure. Unfortunately, the methods for saliva sampling, preparations, and analysis varied across previous studies, which might affect the analytical results. However, limited research was conducted to study the efficacy of using saliva samples to monitor human exposure to metals. The purposes of this study were two aspects. The first was to develop and validate an analytical method for the determination of titanium, chromium, arsenic, cadmium, and lead in human saliva using inductively coupled plasma mass spectrometry (ICP-MS). Tests were performed to evaluate the effects of sample preparation procedures on the measurement results of salivary metal concentrations to optimize the analytical method. Second, 20 healthy adult subjects were recruited for saliva collection, and the newly developed method mentioned above was applied to determine the concentrations of metals in the saliva samples. Meanwhile, a structured questionnaire was administered to survey the study subjects' demographic information, lifestyle, food consumption frequency, and oral hygiene. The concluded preparation procedures for the analysis of the study metals in saliva samples are as follows. Artificial saliva was applied for matrix control to establish the calibration curves. The saliva samples were first centrifuged at 3500 rpm (≅1000 xg) for 10 minutes, and then the supernatant of saliva was stored in a 4°C refrigerator and analyzed within 5 days after sampling. Each saliva supernatant sample was diluted 5 times in volume with 1% nitric acid, vortexed, and filtered prior to metal measurements. The measurement accuracy of this method for titanium, chromium, arsenic, cadmium, and lead was 84.0%, 102.7%, 107.8%, 116.3%, and 117.8%, respectively. And the precision and reproducibility were both less than 8%. With the newly developed method mentioned above, the average concentrations of salivary titanium, chromium, arsenic, cadmium, and lead in the saliva of the study subjects were determined as 140.2 µg/L, 0.21 µg/L, 0.16 µg/L, 0.03 µg/L, and 0.85 µg/L, respectively. In addition, the analysis results show that the average concentrations of these study metals in the 5 saliva samples collected in sequence within a two-hour period were very close. The method developed in this study requires only simple and reliable sample preparation procedures to accurately determine metal concentrations in saliva. This newly developed method is expected to be used as part of risk assessment tools for human exposure to metals. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90254 |
DOI: | 10.6342/NTU202302889 |
全文授權: | 同意授權(限校園內公開) |
顯示於系所單位: | 環境與職業健康科學研究所 |
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