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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳家揚(Chia-Yang Chen) | |
dc.contributor.author | Wan-Yu Shih | en |
dc.contributor.author | 施宛妤 | zh_TW |
dc.date.accessioned | 2021-06-07T23:59:40Z | - |
dc.date.copyright | 2021-02-23 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2021-02-05 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17176 | - |
dc.description.abstract | 台灣與全球已開發國家隨著醫療的進步,都面臨人口老化的問題,促成了預防保健觀念的推廣。其中重要的一環,即為各式機能性食品的出現。除了類似台灣針對「健康食品」相關法規的規範之外,大部分的保健食品沒有嚴格的安全把關機制。如果製造過程中原物料來源或製程的安全性沒有受到保障,經萃取、濃縮等製程後,有可能導致環境污染物、天然毒素、農藥、加工副產品等殘留,造成身體暴露到有害物質、並引起健康風險。 此篇研究以之前藉由血液樣本廣篩建立之技術平台,進一步針對中藥草本類保健食品基質優化,找出最適合之樣本前處理方式,並使用液相層析/高解析串聯式質譜儀進行非標的物分析。雞精、蜆精在本研究為液體基質的代表基質,用以比較兩種前處理方法,包括QuEChERS搭配Oasis PRiME HLB萃取匣及固相萃取(SPE)。固態基質部分,則選定五味子芝麻素、金盞花黑醋栗葉黃素基質用於比較兩種前處理方法,包括QuEChERS搭配Oasis PRiME HLB萃取匣及QuEChERS搭配EMR-lipid tube。樣本通過安捷倫超高效液相層析搭配飛行時間串聯式質譜儀(UHPLC-Q-TOF MS),以正電荷電灑游離(positive electrospray ionization, ESI+)以及負電荷電灑游離(positive electrospray ionization, ESI-)分開分析,進行全離子掃描。以其溶劑使用量、樣本通量、可鑑定波峰數、色素處理能力、訊號強度及穩定性以及方法的萃取效率,對製備方法進行優化。並在最後選擇QuEChERS搭配Oasis PRiME HLB萃取匣作為液態樣本及固態樣本的優先化的方法。 使用QuEChERS搭配Oasis PRiME HLB萃取匣的步驟,先以QuEChERS的方法進行初步的萃取,而後藉由將上層液通過Oasis PRiME HLB萃取匣來達到淨化基質的效果。在溶劑使用的評估上,此優先化前處理方始處理單一樣本只需花費21 mL的溶劑,符合綠色化學的觀念。再者,方法的通量是篩查研究中十分重要的面向,此方法的操作容易、步驟簡單,單一樣本進行前處理的通量落在10.6分鐘,可以在短時間內完成大量樣本的前處理步驟。此外,為評估此方法保留小分子化合物的能力,本研究針對雞精與蜆精及綠茶萃取膠囊與靈芝萃取膠囊進行可供鑑定波峰數(molecular feature)的測試。由結果可知使用QuEChERS搭配Oasis PRiME HLB萃取匣進行前處理,四個基質在ESI+可以偵測到多至1200的波峰數,而在ESI-約落在200-350個之間,最多可多至將近800個波峰數。在液態樣本方法的比較上,使用QuEChERS搭配Oasis PRiME HLB萃取匣進行前處理的可供鑑定波峰數總數較以SmartPrep Automated Cartridge Extractor II進行固相萃取多200個以上;而在固態樣本方法的比較上,使用QuEChERS搭配Oasis PRiME HLB萃取匣進行前處理的可供鑑定波峰數總數則比QuEChERS搭配EMR-lipid tube略低,兩者皆可達到1500個以上。此外,我們也選用五味子芝麻素及金盞花黑醋栗葉黃素為基質,評估此方法的色素處理能力。結果顯示,相較於EMR-lipid tube,Oasis PRiME HLB的淨化較能有效移除樣本中的色素,減少保健食品中色素對於分析的干擾。 在訊號強度的評估,則是在雞精及蜆精與五味子芝麻素及金盞花黑醋栗葉黃素,以前添加方式(pre-spiked)添加12種待測物,包含ESI+的Diethyl phthalate (DEP)、Benzyl butyl phthalate (BBP)、Di(2-ethylhexyl) phthalate (DEHP)、Di-n-octyl phthalate (DNOP) 以及ESI-的Perfluorohexanoic acid (PFHxA)、Perfluorooctanoic acid (PFOA)、Perfluorononanoic acid (PFNA)、Perfluorodecanoic acid (PFDA)、Perfluoroundecanoic acid (PFUnDA)、Sodium Perfluorohexanesulfonate (PFHxS)、Perfluorooctanesulfonic acid (PFOS)及Perfluorododecanoic acid (PFDoDA),以波峰面積(peak area)測試此方法之訊號穩定度,以及藉由前添加(pre-spiked)樣本與後添加(post-spiked)樣本測試其萃取效率。液態樣本在ESI+下,peak area都落在104-106,相對標準偏差(relative standard deviation,%RSD)則落在3.3%-12.1%之間;ESI- peak area則落在104-105,%RSD落在3.7%-13.6%之間,並且萃取效率在ESI+及ESI-中皆在50%上下;固態樣本在ESI+下,peak area都大多落在104-105,%RSD落在2.5%-15.8%之間,而ESI- peak area則落在104-105,%RSD落在0.9%-17.6%之間,萃取效率在ESI+及ESI-皆約落在50%,金盞花黑醋栗葉黃素中的PFOA、PFNA、PFDA、PFUnDA、PFHxS、PFOS的萃取效率在ESI-的萃取效率更可以高至70%。此結果顯示經由QuEChERS搭配Oasis PRiME HLB萃取匣前處理過後,各化合物的訊號強度穩定,代表此方法可以穩定地萃取不同質量範圍的小分子有機物,更可以藉由萃取效率的結果得知此前處理方法在液態及固態基質中,大多皆可以保留50%以上不同質量範圍的小分子有機物,並以金盞花黑醋栗葉黃素的萃取效率最佳。 本研究實測十二種保健食品樣本,以QuEChERS搭配Oasis PRiME HLB萃取匣進行樣本前處理,搭配UHPLC Q-TOF MS進行樣本分析,並與六個資料庫進行篩查比對,包含Agilent廠商建立之黴菌毒素資料庫(Mycotoxins)、農藥資料庫(Pesticides)、藥物與毒物資料庫(Forensic and Toxicology)、畜產用藥資料庫(Veterinary)及水體環境新興污染物資料庫(Water)以及一個本研究團隊自建高度關注物質資料庫(Concerned Chemicals)。本研究發現:辨識頻率較高的化合物大多為藥物與毒物如gentisic acid 、valdetamide等等,比較不屬於一般會用於保健食品中的成分,意味著日常使用的藥物、香料等物質會存在保健食品當中而可能影響到人體健康。本篇研究藉由此快速、穩定、簡易、安全的QuEChERS方法優化以運用到保健食品的基質,以利日後對市面上保健食品潛在污染物進行快速篩查及檢視,為人們攝取的保健食品進行安全把關。 | zh_TW |
dc.description.abstract | Many developed countries, including Taiwan, are facing population aging problems, and disease prevention and health promotion are getting more attentions. Nutraceuticals are wildly used in Taiwan due to its potential to improve health. Although most of the ingredients included in nutraceuticals are natural, there are insufficient safety data regarding trace contaminants and residues that may lead to undesirable health effects. Therefore, comprehensive screening of nutraceuticals is warranted to identify potential hazardous compounds. Chicken extracts and clam extracts were used as the representative liquid matrixes for comparing two sample preparation methods, which were QuEChERS with the cleanup of Oasis PRiME HLB cartridges and solid-phase extraction (SPE). Regarding solid matrixes, sesamin with schisandra extracts and blackcurrant anthocyanins with lutein were used for comparing two sample preparation methods, including QuEChERS with the cleanup of either EMR-lipid tubes or Oasis PRiME HLB cartridges. Samples were analyzed by Agilent UHPLC-Q-TOF MS at both positive and negative electrospray ionization (ESI) with All Ions mode. The optimization of sample preparation methods was based on their used solvent amount, throughput, numbers of molecular features, and the ability to remove pigments in samples. QuEChERS with the cleanup of Oasis PRiME HLB cartridges was suggested as the best method for both liquid and solid matrixes. For QuEChERS with the cleanup of Oasis PRiME HLB cartridges, samples are extracted with QuEChERS first and then cleaned up using the Oasis PRiME HLB cartridges. Only 21 mL solvents were needed for preparing one sample, which meets the principals of Green Chemistry. Also, the extraction process for one sample took only an average of 10.6 minutes. We used the numbers of identified peaks after molecular features extraction (MFE) to determine the ability of this method to keep small molecules in liquid matrixes (chicken extracts and clam extracts) and solid matrixes (green tea extracts and Ganoderma lucidum extracts). We detected up to about 1200 molecular features in ESI+ and up to about 800 molecular features in ESI-. For liquid matrixes, QuEChERS with the cleanup of Oasis PRiME HLB cartridges saved more than 200 molecular features than SPE did. For solid matrixes, QuEChERS with the cleanup of both Oasis PRiME HLB cartridges and EMR-lipid tubes could save up to 1500 molecular features. As a result, we considered QuEChERS with the cleanup of Oasis PRiME HLB cartridges as the best method to keep small molecules for both liquid matrixes and solid matrixes. In the study, we also compared QuEChERS with the cleanup of Oasis PRiME HLB cartridges to QuEChERS with the cleanup of EMR-lipid tube to determine the remove of pigments from solid matrixes (sesamin with schisandra extracts and blackcurrant anthocyanins with luteins). We found that QuEChERS with the cleanup of Oasis PRiME HLB cartridges was better for eliminating pigments from the matrixes. To confirm whether QuEChERS with the cleanup of Oasis PRiME HLB cartridges is stable enough to be served as a sample preparation method for suspect screening analysis, 12 analytes, including Diethyl phthalate (DEP), Benzyl butyl phthalate (BBP), Di(2-ethylhexyl) phthalate (DEHP) and Di-n-octyl phthalate (DNOP) for ESI+ and Perfluorohexanoic acid (PFHxA), Perfluorooctanoic acid (PFOA), Perfluorononanoic acid (PFNA), Perfluorodecanoic acid (PFDA), Perfluoroundecanoic acid (PFUnDA), Sodium Perfluorohexanesulfonate (PFHxS), Perfluorooctanesulfonic acid (PFOS) and Perfluorododecanoic acid (PFDoDA) for ESI-, were added in liquid matrixes (chicken extracts and clam extracts) and solid matrixes (sesamin with schisandra extracts and blackcurrant anthocyanins with lutein) using both pre-spiked and post-spiked approaches with four duplicates. Stability was evaluated using the relative standard deviation (%RSD) calculated based on the peak areas of the pre-spiked samples (four duplicates). Extraction efficiency was evaluated by comparing the peak areas of pre-spiked samples to post-spiked samples. In liquid matrixes, the %RSD of peak areas of 12 analytes was 3.3%-12.1% in ESI+ and 3.7%-13.6% in ESI-. The extraction efficiencies of all the analytes in the liquid matrixes were at about 50%. As for solid matrixes, the %RSD of peak areas of analytes was 2.5%-15.8% in ESI+ and 0.9%-17.6% in ESI-. The extraction efficiencies of analytes in ESI+ were at about 50% while those in ESI- were at about 70%. We considered that QuEChERS with the cleanup of Oasis PRiME HLB cartridges is a stable method for both liquid and solid matrixes with the extraction efficiencies of about 50%. In this study, we further implemented QuEChERS with the cleanup of Oasis PRiME HLB cartridges in 12 nutraceuticals. We analyzed the extracted samples using Agilent UHPLC-Q-TOF MS at both ESI+ and ESI-, and we conducted suspect screening based on six libraries, including five libraries provided by Agilent (Mycotoxins, Pesticides, Water, Veterinary Drugs and Forensic/Toxicology) and one self-built concerned chemicals libraries in Personal Compound Databases and Library (PCDL). We found that compounds with detection frequencies over 50%, such as gentisic acid and valdetamide, were mostly from the Forensic and Toxicology library. These compounds were detectable in our selected 12 nutraceuticals potentially and further investigation of their potential health effects is needed. In conclusion, this study found that QuEChERS with the cleanup of Oasis PRiME HLB cartridges is suitable for sample preparation of nutraceuticals for conducting suspect screening of chemical contaminants using UHPLC Q-TOF MS. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T23:59:40Z (GMT). No. of bitstreams: 1 U0001-0502202100250500.pdf: 3683319 bytes, checksum: b66980b9be1ace86751aa33f6e67b835 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 致謝 I 中文摘要 II 英文摘要 V 目錄 I 圖目錄 I 表目錄 II 第一章、 前言 1 1.1 研究緣起 1 1.2 保健食品及潛在隱憂 2 1.3 質譜技術探討(mass spectrometry) 3 1.4 非標的物分析(non-target analysis)與潛在物篩查(suspect screening) 5 1.5 多重殘留物質分析檢測之樣品之前處理方式 6 1.6 研究目的 8 第二章、 研究方法 9 2.1 試劑與材料 9 2.2 儀器使用 11 2.3 樣本收集 11 2.4 前處理方法 12 2.5 層析方法 13 2.6 四級柱飛行時間質譜儀(Q-TOF MS) 13 2.7 分析軟體與資料處理流程及規範 14 2.7.1 個人化合物數據資料庫管理B.07.00(PCDL) 14 2.7.2 化合物篩查流程及資料處理流程與規範 15 2.8 品質保證及品質管制 16 2.8.1 品質保證(QA) 16 2.8.2 品質管制(QC) 16 第三章、 結果與討論 19 3.1 優化樣本前處理方法 19 3.1.1 溶劑使用 20 3.1.2 樣本處理通量 21 3.1.3 可供鑑定波峰數(molecular feature) 23 3.1.4 色素處理能力 25 3.1.5 前處理方法之選擇 26 3.2 QuEChERS萃取搭配Oasis PRiME HLB萃取匣的方法測試 27 3.2.1 訊號強度 27 3.2.1.1 液態樣本中的訊號強度及穩定性 28 3.2.1.2 固態樣本中的訊號強度及穩定性 29 3.2.2 方法萃取效率 30 3.2.3 方法測試的品質管理與品質控制(QA/QC) 31 3.3 實際樣本運用前處理方法進行篩查分析 32 第四章、 結論與未來展望 37 參考資料 39 附圖 47 附表 69 | |
dc.language.iso | zh-TW | |
dc.title | 建立超高效液相層析搭配飛行時間串聯式質譜平台篩查保健食品中化學污染物 | zh_TW |
dc.title | Platform for Screening of Chemical Contaminant in the Nutraceuticals Using Ultra-high Performance Liquid Chromatography/Quadrupole Time-of-flight Mass Spectrometry | en |
dc.type | Thesis | |
dc.date.schoolyear | 109-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭錦樺(Ching-Hua Kuo),陳宏彰(Hong-Jhang Chen),呂廷璋(Ting-Jang Lu) | |
dc.subject.keyword | 超高效液相層析飛行時間串聯式質譜儀,篩查,保健食品,電灑游離法, | zh_TW |
dc.subject.keyword | suspect screening,functional food,UHPLC-Q-TOF MS,QuEChERS,electrospray ionization, | en |
dc.relation.page | 84 | |
dc.identifier.doi | 10.6342/NTU202100554 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2021-02-08 | |
dc.contributor.author-college | 公共衛生學院 | zh_TW |
dc.contributor.author-dept | 環境與職業健康科學研究所 | zh_TW |
顯示於系所單位: | 環境與職業健康科學研究所 |
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