201231136 六、發明說明： 【發明所屬之技術領域】 本發明係與一種萃取裝置有關’特別是關於一種液相 萃取裝置以及應用液相萃取裝置的樣品前處理方法。 【先前技術】 樣品的前處理具有減少基質效應、提高分析物含量以 及提昇分析靈敏度等作用，因此前處理技術為樣本分析中 最重要的一個步驟’例如中華民國第1284198號發明專利 便提供了一種快速化學物質檢測預處理之方法及設備，以 減少樣品前處理之時間。 一般而言，樣品前處理技術區分為固相萃取(solid pha se extraction)以及液相萃取(liquid phase extraction)，其中 固相萃取可以減少或避免有機溶劑的使用，但是成本較為 昂貴，並且會造成許多廢棄物；液相萃取則使用較為大量 的有機溶劑’而容易造成環境污染，但操作簡單，並且成 本較為低廉。 為了解決舊有萃取方法的問題，具有簡便、低劑量、 萃取時間短，並且可與其他分析儀器結合，進行微量分析 等特點的微量萃取法，便成為近年來重要的研究方向，其 中又以液相微萃取法(liquid phase microextraction)的開發 最為受到重視。 1996年Liu與Dasgupta研發出單滴微萃取法(drop_in-drop system) ’運用液液萃取中分配作用之觀念，將與水不 201231136 互不相溶的有機溶劑液滴，放置在一個流動的大液滴中， 不斷進行萃取的技術。（Liu，H.，Dasgupta，P. K. (1996).201231136 VI. Description of the Invention: Field of the Invention The present invention relates to an extraction apparatus, particularly relating to a liquid phase extraction apparatus and a sample preparation method using the liquid phase extraction apparatus. [Prior Art] The pretreatment of the sample has the effects of reducing the matrix effect, increasing the analyte content, and improving the sensitivity of the analysis. Therefore, the pretreatment technique is the most important step in the sample analysis. For example, the invention patent of the Republic of China No. 1284198 provides a kind of invention patent. Rapid chemical detection and pretreatment methods and equipment to reduce sample preparation time. In general, sample pretreatment techniques are divided into solid phase extraction (solid phase extraction) and liquid phase extraction (liquid phase extraction), in which solid phase extraction can reduce or avoid the use of organic solvents, but it is more expensive and causes Many wastes; liquid phase extraction uses a relatively large amount of organic solvent' and is prone to environmental pollution, but it is simple to operate and relatively inexpensive. In order to solve the problem of the old extraction method, the micro-extraction method with simple, low-dose, short extraction time, and can be combined with other analytical instruments for microanalysis has become an important research direction in recent years, among which liquid The development of liquid phase microextraction has received the most attention. In 1996, Liu and Dasgupta developed a drop-in-drop system. Using the concept of partitioning in liquid-liquid extraction, the organic solvent droplets that are incompatible with water not 201231136 are placed in a large flow. In the droplets, the technique of continuous extraction. (Liu, H., Dasgupta, P. K. (1996).
Analytical chemistry in a drop. Solvent extraction in a microdrop. Analytical Chemistry, ¢5(11), 1817-1821.) 同年Jeannot與Cantwell發表新的單滴微萃取技術， 以鐵氟龍棒固定一個有機溶劑液滴；並且將有機溶劑液滴 浸入到由磁石擾拌的轉動水樣溶液中，經過一段時間的萃 取後，注入氣相層析儀分析。（jeann〇t，μ. A.，& Cantwel 1, F. F. (1996). Solvent microextraction into a single dro p. Analytical Chemistry, 68(13), 2236-2240.) 1997年Jeannot與Cantwell進一步改良前述的單滴溶 劑微萃取法，使用微量注射的斜口針尖代替鐵氟龍棒來固 定有機溶劑液滴，並且將有機溶劑液滴浸入水相樣品中進 行萃取’而可以更便利地注入氣相層析儀分析。（Jeann〇t， Μ. Α., & Cantwell, F. F. (1997). Mass transfer characte ristics of solvent extraction into a single drop at the tip of a syringe needle. Analytical Chemistry, 69(2), 235-2 39.) 1998年Ma與Cantwell首度把萃取與反萃取(extracti〇 n/back-extraction)結合在同一步驟中。當分析物屬於鹼性分 子的情況下’在樣品相(donor phase)pH值為13時，分析 物為中性分子，而可將分析物分配萃取至有機相(〇rganic phase);在接收相(acceptor phase) pH值為2.1時，分析物 則會質子化，進而將使分析物反萃取至接收相。（Ma, M., 201231136 & Cantwell, F. F. (1999). Solvent microextraction with s imultaneous back-extraction for sample cleanup and prec oncentration: preconcentration into a single microdrop. A nalytical Chemistry, 70(18), 3912-3919.) 然而’ Ma與Cantwell的萃取裝置進行單滴溶劑微萃 取時，由於液滴不易維持，並且容易受到樣品中氣泡和基 質微小粒子的影響’而可能在萃取過程中掉落，因此無法 提高萃取過程的溫度及攪拌速率等條件。 鲁 1999 年 Pedersen-Bjergaard 與 Rasmussen 發表中空纖 維液相微萃取法(hollow fiber liquid-phase microextractio π ’ HF-LPME) ’ 運用多孔性聚丙稀(p〇iypr〇piyene，pp)材 質的中空纖維管來保護溶劑，以改善上述缺點。（pedersen_ Bjergaard, S., & Rasmussen, K. Ε. (1999). Liquid-liquid-liquid microextraction for sample preparation of biologic al fluids prior to capillary electrophoresis. Analytical Ch • emistry, 7/(14), 2650-2656.) 2002年Hou與Lee進而提出以微量注射針結合一段 中空纖維管進行液相微萃取，萃取後直接以注射針抽取中 空纖維管内的溶劑注入氣相層析儀分析，且將中空纖維液 相微萃取法中有分為兩液相(two-phase)與三液相(three-pha se)兩種模式。先將中空纖維管之孔洞中充滿有機溶劑，用 微量注射針將有機溶劑（兩液相）或者鹽酸水溶液(三液相） 從一端注入中空纖雄管内，再浸入樣品中萃取，隨後反萃 取至鹽酸中取得分析物。（H〇u，L” & Lee，Η. K. (2003). 201231136Analytical chemistry in a drop. Solvent extraction in a microdrop. Analytical Chemistry, ¢ 5(11), 1817-1821.) In the same year, Jeannot and Cantwell published a new single-drop microextraction technique to fix an organic solvent droplet with a Teflon rod. And immersing the organic solvent droplets into the rotating water sample solution disturbed by the magnet, and after a period of extraction, injecting into a gas chromatograph for analysis. (jeann〇t, μ. A., & Cantwel 1, FF (1996). Solvent microextraction into a single dro p. Analytical Chemistry, 68(13), 2236-2240.) In 1997, Jeannot and Cantwell further improved the aforementioned Single-drop solvent micro-extraction method, using micro-injected oblique tip to replace Teflon rod to fix organic solvent droplets, and immersing organic solvent droplets in aqueous phase for extraction', which can be more conveniently injected into gas chromatography. Instrument analysis. (Jeann〇t, Μ. Α., & Cantwell, FF (1997). Mass transfer characte ristics of solvent extraction into a single drop at the tip of a syringe needle. Analytical Chemistry, 69(2), 235-2 39 .) In 1998, Ma and Cantwell first combined extraction and back extraction (extracti〇n/back-extraction) in the same step. When the analyte belongs to a basic molecule, 'when the pH of the donor phase is 13, the analyte is a neutral molecule, and the analyte can be extracted and extracted into the organic phase (〇rganic phase); (acceptor phase) When the pH is 2.1, the analyte will be protonated, which in turn will back-extract the analyte to the receiving phase. (Ma, M., 201231136 & Cantwell, FF (1999). Solvent microextraction with s imultaneous back-extraction for sample cleanup and prec oncentration: preconcentration into a single microdrop. A nalytical Chemistry, 70(18), 3912-3919. However, 'Ma and Cantwell's extraction device for single-drop solvent microextraction, because the droplets are difficult to maintain, and are susceptible to bubbles and matrix microparticles in the sample' may fall during the extraction process, so the extraction process cannot be improved Conditions such as temperature and stirring rate. In 1999, Pedersen-Bjergaard and Rasmussen published hollow fiber liquid-phase microextractio π 'HF-LPME' using hollow fiber tubes made of porous polypropylene (p〇iypr〇piyene, pp). The solvent is protected to improve the above disadvantages. (Pedersen_ Bjergaard, S., & Rasmussen, K. Ε. (1999). Liquid-liquid-liquid microextraction for sample preparation of biologic al fluids prior to capillary electrophoresis. Analytical Ch • emistry, 7/(14), 2650- 2656.) In 2002, Hou and Lee further proposed liquid microextraction with a microinjection needle combined with a hollow fiber tube. After extraction, the solvent in the hollow fiber tube was directly extracted by injection needle into a gas chromatograph for analysis, and the hollow fiber solution was The phase microextraction method is divided into two modes: two-phase and three-phase. First, the pores of the hollow fiber tube are filled with an organic solvent, and an organic solvent (two liquid phase) or an aqueous hydrochloric acid solution (three liquid phase) is injected into the hollow fiber male tube from one end with a micro injection needle, and then immersed in the sample for extraction, followed by back extraction to The analyte was taken in hydrochloric acid. (H〇u, L" & Lee, Η. K. (2003). 201231136
Dynamic three-phase microextraction as a sample prepa ration technique prior to capillary electrophoresis, Analyt ical Chemistry, 75(11), 2784-2789.) 相較於單滴微萃取法’當兩者之萃取溶劑體積相同 時，中空纖維管可提供有機溶劑與樣品較大接觸面積，並 且溶劑液滴因中空纖維管支撐保護之故，不容易逸失於水 樣申。此外’中空纖維管可固定於微量注射針尖，配合注 射針式幫浦推拉注射針推桿，來提高分析物的萃取效率。(J iang, X., 〇h, S. Y., & Lee, Η. K. (2005). Dynamic liq uid-liquid-liquid microextraction with automated moveme nt of the acceptor phase. Analytical Chemistry, 77(6), 16 89-1695.) 2004年Jang與Lee又發展出溶劑棒微萃取法(s〇lvent bar microextraction)’在適當長度的中空纖維管中注入萃取 用溶劑後，將中空纖維管兩端密封形成一溶劑棒，再將溶 劑棒置入溶液中隨磁石轉動，以進行分配作用而達到萃取 效果。（X. Jang，& Η. K. Lee (2004). Solvent bar micro extraction. Analytical Chemistry, 76(18), 5591-5596 ) 此方法相較於單滴微萃取與靜態的中空纖維管液相萃 取’溶劑棒微萃取法藉由增加其界面接觸面積及攪動速率 來達到更高的萃取效率。 2004年Schellm、Hauser與P〇pp將中空纖維管的容 量再擴大，由於可叫載大體積的溶劑，搭配氣相層析儀 大體積注射，因此能夠有效提昇分析物之偵測極限。（驗 201231136 llin, M., Hauser, B., & Poppa, P. (2004). Determination of organophosphorus pesticides using membrane-assisted solvent extraction combined with large volume injection-gas chromatography-mass spectrometric detection. Journa l of Chromatography A, 1040(2), 251-258.) 2008年Pan等人則使用PCR離心管做為材料進行三 液相萃取，先將接收相放入PCR離心管前端，再將有機相 放入並覆蓋接收相’隨後用樣品相將PCR離心管填滿，再 將其插入裝滿樣品相的樣品瓶中，躺置進行三液相萃取。(P an, W., Xu, H., Cui, Y., Song, D., & Feng, Y. Q. (200 8). Improved liquid-liquid-liquid microextraction method and its application to analysis of f〇ur phenolic compou nds in water samples. Journal of Chromatography A, 12 03(1), 7-12.) 綜合以上各類液相萃取技術，單滴溶劑微萃取法因為 微滴不易維持，所以發展出利用中空纖維管用來支撐及保 護萃取溶劑’並且增加萃取表面積與溶義敎度。由於 中空纖維管表面的孔洞具有選，謂大分子阻擋而使 大分子無法進人萃取溶劑，因此可降低樣品中的基質干 擾，而有助於顧在複雜基質樣品中有機化合物的分析。 但是中空_管躲祕縣親品村齡造成容易汗 染及孔洞阻塞關題，使得中空纖㈣-般皆無法重複使 用。此外’使用過於微小體積的有機相，亦容易產生溶離 及揮發性損失關題而使可應用之有機相麵受限制。 201231136 【發明内容】 本發明之主要目的在於提供一種液相萃取裝置以及應 用前述液相萃取裝置的樣品前處理方法，其中該液相萃取 裝置之成本低廉、可以重複使用，並且應用該液相萃取裝 置的樣品前處理方法具有較簡單之操作步驟。 為達成則述目的，本發明提供一種液相萃取裝置，包 3有第谷器、一第一容器以及一管件，其中該第-容 器容置於該第-容器，且該第二容器具有—盛置部以^ 身部，該盛置部可盛裝溶液，該身部具有一連通該第二容 器内外之開口；該管件容置於該第二容器。 本發明另外提供一種應用前述液相萃取裝置的樣品前 處理方法’包含有下列步驟：⑻加人有機相於盛裝有樣品 相之該第一容器；（b)置放該管件於該第二容器，並且將接 收相加入該第二容器；（C)將該管件與盛裝有接收相之該第 二容器一同置入該第一容器’使有機相經由該開口進入該 第二容器；以及(d)從該管件内取出接收相。 據此，本發明所提供之液相萃取裝置及應用該液相萃 取裝置的樣品前處理方法，其結構簡單便於製作，並且經 過清洗後便能重複使用，操作上也十分穩定及便利。 【實施方式】 為了詳細說明本發明之構造及特點所在，茲舉以下之 較佳實施例並配合圖式說明如后，其中： 201231136 第一圖為本發明一較佳實施例之立體圖； 第二圖為本發明一較佳實施例之剖視圖； 第三圖為本發明一較佳實施例中，第一容器加入磁性 攪拌子、樣品相以及有機相，第二容器加入接收相並置入 S'件’且第二容器尚未置入第一容器内之示意圖； 第四圖為本發明一較佳實施例中，第一容器加入磁性 攪拌子、樣品相以及有機相，第二容器加入接收相並置入 管件，且第二容器置入第一容器内之示意圖； 第五圖為本發明一較佳實施例中，從管件取出接收相 之示意圖； 第六圖為尿液樣本添加曱酚標準品溶液萃取前後與甲 酚標準品溶液之比較分析圖，其中標號(1)為尿液樣本添加 IfAg/ml之曱紛標準品溶液，並且未經過萃取之實驗圖譜；標 號(2)為水溶液添加ιμδ/ιη丨之曱酚標準品溶液之實驗圖譜；標 號(3)為尿液樣本添加1 之甲酚標準品溶液，並且經過萃 取之實驗圖譜； 第七圖為尿液樣本添加苄基硫醇酸標準品溶液萃取前 後與苄基硫醇酸標準品溶液之比較分析圖，其中標號(1)為 尿液樣本添加1μβ/πι1之苄基硫醇酸標準品溶液，並且未經過 萃取之實驗圖譜；標號(2)為水溶液添加ipg/mi之苄基硫醇酸 標準品溶液之實驗圖譜;標號⑶為尿液樣本添加1 μβ/ιη1之苄 基硫醇酸標準品溶液’並且經過萃取之實驗圖譜；以及 第八圖為管柱後注入測試之比較分析圖，其中標號(1) 為尿液樣本添加〇·2Μ磷酸水溶液，並且經過萃取之實驗 201231136 圖譜；標號(2)為甲酸鹽類水溶液之實驗圖譜；標號(3)為尿 液樣本添加G.2M雜水溶液，並且未經料取之實驗圖 譜。 如第-圖與第二圖所示，本發明一較佳實施例所提供 之液相萃取裝置10包含有一第一容器2〇、一第二容器3〇 以及一管件40。 該第一容器20之一端封閉，另一端具有一容器口 22。 該第一今器30穿過该容器口 22而容置於該第一容器 2〇。該第二容器30具有一盛置部32以及一身部％，該盛 置部32之-端賴，散該盛置部32之雜朝封閉端方 向逐漸收縮而概呈圓錐狀結構；對照第一圖與第二圖所 示’該身部34之壁面相對開設有二開口 36。該第二容器3 〇更具有-ϋ定件38，該固定件38蓋合於該身部34。 該管件40穿過該固定件38而容置於該第二容器3〇， 藉由該固定件38使該管件38維持在直立狀態，且該管件 40之一端伸入該第二容器3〇而位於該盛置部32，該管件 40之另一端可開合地蓋合有一蓋件42。 藉由上述結構與技術特徵，以下進一步說明應用該液 相萃取裝置10的樣品前處理方法。 首先，依照所欲萃取之分析物穩類分別製備樣品相 D、有機相〇以及接收相Α溶液，其中樣品相D、有機相 〇以及接收相Α溶液係依照不同樣本、分析物以及實驗設 叶的需要而有不同的配置方法。如第三圖所示，上述溶液 製備完成之後，先將樣品相D與一攪拌子24加入該第一 201231136 容器20後，再加入有機相0。 然後將該管件40穿過該固定件38而置放於該第二容 器40,並且將接收相Α加入該第二容器3〇,此時接收相A 盛裝於該盛置部32 ’使接收相A之液面低於該開口 36。 如第四圖所示，接著將該管件40以及盛裝有接收相A 之第二容器30 —起放入該第一容器20，並且將該第二容 器30放置至該開口 36低於有機相〇之液面，使得有機相 Ο經由該開口 36流入該第二容器30而與接收相A相互接 • 觸後，蓋上該蓋件42，攪拌一段時間進行萃取。 萃取完成之後，如第五圖所示，便可打開該蓋件42， 從該管件40内取出接收相A。取出接收相a並且經過中 和之後，便可將中和後之接收相A輸入檢測儀器中分析。 據此，本發明所提供之液相萃取裝置及應用該液相萃 取裝置的樣品前處理方法，可同時進行萃取及反萃取，且 該液相裝置之結構簡單便於製作，經過清洗後便能重複使 • 帛，而可降低不必要的耗材與成本，操作上也十分便利簡 單。另一方面，本發明所提供之液相萃取裝置將接收相盛 置並且保護在第二容器内，即使有機相溶液因為萃取時間 而揮發或者是溶失於水中時，樣品相也無法與接收相^ 通，因此在萃取過程中具有較佳的穩定度，也能夠增加有 機相中有機溶劑的選擇性。 以下藉由一實驗例進一步說明本發明。 <實驗例〉 f苯(toluene methyU)en職)是一種芳香環類的破 201231136 氫化合物，廣泛使用於工業的有機溶劑，無色，微甜且具 有刺激性的可燃性液體。自然界中的曱苯主要來源是原油 燃料及香膠樹中’甲苯的產生，主要經由石化工業製造過 程而來。然而曱苯常被用於化學產品中，如油漆、膠水及 稀釋劑、製造塗料、塗料稀釋劑、黏著劑及橡膠，也被用 於印刷及皮革鞣製過程等。 曱苯具有脂溶性特性，很快會經由血液進入中樞神經 系統。吸食後產生幻覺及欣快感，且對外界刺激極為敏感， 易衝動而產生行為上的偏差，若吸食過量隨著血液中濃度 籲 增加會產生精神錯亂、運動失調、無方向感等中樞抑制行 為°在人類’急性口服曱苯中毒則會造成有口咽灼熱的感 覺、胃痛、腹部疼痛、咳血、噁心、嘔吐，及虛弱的感覺。 其他可能的症狀包括暈眩、欣快感、胸悶、步履蹣跚。更 嚴重則包括視覺糢糊、顫抖、呼吸淺而快速、痲痺癱瘓、 無意識及痙攣，大部份的症狀都為中樞神經系統毒性的表 現，所以監控其甲苯曝露量有其重要性。 在我國相關的研究報告中提出，生物偵測是量測生物 _ 檢體中的有害物及其代謝物，此受檢測物質稱為生物指標 (biomarker)，由於它可以彌補環境偵測方面的不足近年 來已有許多研究在使用。生物指標從化學物質的暴露到疾 病產生的過程中，需要經過許多中間步驟，其間有許多生 物指標可用來評估人體暴露量與損傷情況。一般將生物指 標分成3類，分別為暴露生物指標（exp〇sure bi〇marke r)、健康效應生物指標（health effect biomarker)與易感性 12 201231136 生物指標（susceptibility biomarker)，可以再細分成5類， 各別為内在劑量、生物有效劑量（biological effective dos e)、早期效應（early response)、結構或功能上的改變（ait ered structure/function)與易感性生物指標；内在劑量與生 物有效劑量被分類為暴露生物指標，早期效應與結構或功 能上的改變為健康效應指標，各有不同的生理意義。 在甲苯代謝的文獻中，馬尿酸(hippuric acid)與對位曱 酚(para-cresol)為人體胺基酸的主要代謝物，容易受到飲食 及其他有機溶劑共同暴露而影響甲苯在人體内的代謝，因 而影響到尿液中對位甲酚與馬尿酸的濃度，因此馬尿酸與 對位曱酚較不適合作為低濃度甲苯暴露指標物。因此尿中 代謝物鄰位曱紛(ortho-cresol)與苄基硫醇酸(benzylmercapt uric acid，BMA)，是曱苯較佳之暴露指標物。 萃取裝豎之製作 本實驗中之萃取裝置使用0.4ml以及1.5ml的離心管 所組裝而成’ 一般離心管為聚丙稀(p〇lypr〇pylene)材質，由 於本實驗涉及較為激烈的酸驗溶液，因此選用矽處理過的 離心管(pre-lubricated tube/siliconized tube)。 在1.5ml離心管的兩侧管壁剪出一個適當開口，並且 在管蓋上鑽洞’將0.4ml離心管的底端切除後穿入1.5ml 的離心管管蓋中，再以超音波清洗器震盪清洗三十分鐘， 放置數分鐘後乾燥即可使用。 樣品之配詈 收集未曾暴露於曱笨溶劑之身體健康成人尿液，並將 13 201231136 尿液以3000g的轉速離心五分鐘後，取出上清液備用。 精秤鄰位甲紛;(para-cresol 99%，購買自 Alfa Aesar)l Omg放置於l〇ml的量液瓶中，以去離子水稀釋至標線後， 均勻混和而得濃度lOOOpg/ml之鄰位甲酚溶液。重複前述 步驟’可得濃度lOOOpg/ml之對位甲紛(ortho-cresol 99%， 購貝自 Alfa Aesai)以及节基硫醇酸(benzylmercapturic aci d 99%，購買自 Alfa Aesar)溶液。 將濃度lOOOpg/ml之鄰位甲酚、對位甲酚以及苄基硫 醇酸稀釋並且混合成為I各自濃度分別為250pg/ml之標準混 合溶液後，取出40μβ加入尿液樣本至i〇m卜而成為各自濃度 分別為lpg/ml之混合標準添加樣品。 萃取裝晉之細厶乃萃 於樣品瓶(10ml)中放入約6.0ml的樣品相及磁性攪拌子 後’於樣品瓶中加入1.5ml有機相，其中樣品相依照實驗設計 上之需求而有不同配方，有機相則是由甲苯(t〇luene，購買自 Merck ’德國)與乙醚(dithyl ether，購買自Merck，德國)以一 比一體積混合而成。 於先前製作完成的離心管組件内加入〇 4ml、〇 1M氫氧化 納溶液作為接收她，雜心管崎放人裝有樣品相的樣品瓶 中，在50 C環境下攪拌約15〇分鐘進行萃取，萃取完畢後利 用注射針將接收相取出巾和後，注人檢測儀H中分析。 件與參數設定 在液相層析儀串連質譜儀的分析中，液相層析儀部份 使用的疋咼效能液相層析系統(Waters ACQUITY Ultraper 201231136 formance LC system，購買自美國），分離管柱則是將phe i nomenex Kintex Cl8(3.5μιη’2.1x30mm)與 Supelico Diphenyl(2. :6μιη，2.1x150mm)串聯，並且在管柱溫度4(rc的環境下以〇j 5ml/min的流速進行等梯度沖提，動相組成為水與乙腈(acet〇ni • 1 1 ，GR，購買自Merck，德國)以60與40的體積比例調配 而成。 質譜儀(Waters TQD mass spectrometer，購買自英國） 4伤以負離子模式彳貞測(negative i〇n mode)，在全掃描模 式(full-scan mode)中，以質量範圍為80〜35〇m/z進行偵 測。先改變毛細管出口端電壓之後，再改變碰撞室能量， 便可以得到欲分析之化合物在不同能量下的斷裂離子訊號 強度變化。並且在tune模式下，依序改變毛細管出口端之 電壓後’在子代離子掃描模式(Daughters ion scan，Dau-S can)中將分析前趨離子(precurs〇r i〇n，[m_h]·)選出，並依 序改變碰撞室能量值’可得到各離子不同子代離子訊號， 而我們選擇能得到訊號最大的子代離子作為所採用的碰撞 至月匕1。多重反應監控模式(Multiple reaction monitoring) 之參數如下：Dynamic three-phase microextraction as a sample preparation technique prior to capillary electrophoresis, Analyt ical Chemistry, 75(11), 2784-2789.) Compared to single droplet microextraction method, when the extraction solvent volume of the two is the same, hollow The fiber tube can provide a large contact area between the organic solvent and the sample, and the solvent droplets are not easily lost to the water sample due to the support of the hollow fiber tube support. In addition, the hollow fiber tube can be fixed to the micro-injection needle tip, and the injection needle-type push-pull needle can be used to improve the extraction efficiency of the analyte. (Jiang, X., 〇h, SY, & Lee, Η. K. (2005). Dynamic liq uid-liquid-liquid microextraction with automated moveme nt of the acceptor phase. Analytical Chemistry, 77(6), 16 89-1695.) In 2004, Jang and Lee developed a solvent rod microextraction method. After injecting the solvent for extraction into a hollow fiber tube of appropriate length, the two ends of the hollow fiber tube were sealed to form a solvent. The rod is placed in the solution and rotated with the magnet to perform the distribution to achieve the extraction effect. (X. Jang, & Η. K. Lee (2004). Solvent bar micro extraction. Analytical Chemistry, 76(18), 5591-5596) This method is compared to single-drip microextraction and static hollow fiber tube liquid phase. Extraction 'Solvent rod microextraction method achieves higher extraction efficiency by increasing its interface contact area and agitation rate. In 2004, Schellm, Hauser and P〇pp expanded the capacity of the hollow fiber tube. Because it can be called a large volume of solvent, combined with a large volume injection of a gas chromatograph, it can effectively increase the detection limit of the analyte. (Test 201231136 llin, M., Hauser, B., & Poppa, P. (2004). Determination of organophosphorus pesticides using membrane-assisted solvent extraction combined with large volume injection-gas chromatography-mass spectrometric detection. Journa l of Chromatography A, 1040(2), 251-258.) In 2008, Pan et al. used a PCR centrifuge tube as a material for three-liquid extraction. The receiving phase was first placed in the front end of the PCR centrifuge tube, and the organic phase was placed and covered. The receiving phase 'fills the PCR tube with the sample phase, inserts it into the vial filled with the sample phase, and lays it down for a three-phase extraction. (P an, W., Xu, H., Cui, Y., Song, D., & Feng, YQ (200 8). Improved liquid-liquid-liquid microextraction method and its application to analysis of f〇ur phenolic Compo nds in water samples. Journal of Chromatography A, 12 03(1), 7-12.) Combining the above various liquid phase extraction techniques, single-drop solvent micro-extraction method is developed for the use of hollow fiber tubes because the droplets are not easily maintained. To support and protect the extraction solvent' and increase the extraction surface area and solubility. Since the pores on the surface of the hollow fiber tube are selected, that is, the macromolecular blockage prevents the macromolecule from entering the extraction solvent, thereby reducing the matrix interference in the sample and facilitating the analysis of the organic compound in the complex matrix sample. However, the hollow _ tube hides the secret village age, causing easy sweating and hole blocking, so that the hollow fiber (four) - can not be reused. In addition, the use of an organic phase that is too small in volume is also prone to elution and loss of volatility, which limits the applicable organic phase. 201231136 SUMMARY OF THE INVENTION The main object of the present invention is to provide a liquid phase extraction apparatus and a sample pretreatment method using the same as the liquid phase extraction apparatus, wherein the liquid phase extraction apparatus is low in cost, can be reused, and is applied by the liquid phase extraction. The sample preparation method of the device has a relatively simple operation step. To achieve the above, the present invention provides a liquid phase extraction apparatus, the package 3 having a trough, a first container, and a tube member, wherein the first container is housed in the first container, and the second container has - The holding portion is a body portion, and the holding portion can hold a solution, and the body has an opening that communicates with the inside and outside of the second container; the tube member is accommodated in the second container. The present invention further provides a sample pretreatment method for applying the liquid phase extraction apparatus described above, comprising the steps of: (8) adding an organic phase to the first container containing the sample phase; (b) placing the tube in the second container And adding a receiving phase to the second container; (C) placing the tube together with the second container containing the receiving phase into the first container 'to allow the organic phase to enter the second container through the opening; and (d ) take the receiving phase out of the tube. Accordingly, the liquid phase extraction apparatus provided by the present invention and the sample pretreatment method using the liquid phase extraction apparatus are simple in structure, easy to manufacture, and can be repeatedly used after washing, and are also very stable and convenient in operation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In order to explain the structure and features of the present invention in detail, the following description of the preferred embodiments and the accompanying drawings, wherein: FIG. BRIEF DESCRIPTION OF THE DRAWINGS In a preferred embodiment of the invention, a first container is provided with a magnetic stirrer, a sample phase and an organic phase, and a second container is added to the receiving phase and placed in the S' A schematic view of the second container is not placed in the first container; the fourth figure is a first embodiment of the present invention, the first container is added with a magnetic stirrer, a sample phase and an organic phase, and the second container is added to the receiving phase. Schematic diagram of placing a tubular member and placing the second container into the first container; Figure 5 is a schematic view of the receiving phase taken out from the tubular member in a preferred embodiment of the present invention; and Figure 6 is a diagram showing the addition of phenolic standard to the urine sample. Comparative analysis of the cresol standard solution before and after solution extraction, wherein the label (1) is a standard sample solution of IfAg/ml added to the urine sample, and the experimental map is not extracted; label (2) The experimental solution of the solution of ιρ/ιη丨's phenolic standard solution is added to the aqueous solution; the label (3) is the cresol standard solution of the urine sample, and the extracted experimental spectrum is obtained; the seventh figure is the addition of the benzyl group to the urine sample. A comparative analysis of a thiol acid standard solution before and after extraction with a benzyl thiol acid standard solution, wherein the label (1) is a 1 μβ/πι 1 benzyl thiol acid standard solution added to the urine sample, and is not extracted. Experimental map; label (2) is an experimental map of ipg/mi benzyl thiol acid standard solution in aqueous solution; label (3) is added 1 μβ/ιη1 of benzyl thiol acid standard solution for urine sample and extracted The experimental map; and the eighth figure is a comparative analysis of the post-injection test, wherein the label (1) is a urine sample added with a 〇·2Μ phosphoric acid solution, and the extracted experiment 201231136 map; the label (2) is formate An experimental map of an aqueous solution; label (3) is an experimental sample in which a G.2M heteroaqueous solution is added to a urine sample and is not taken. As shown in the first and second figures, a liquid phase extraction apparatus 10 according to a preferred embodiment of the present invention comprises a first container 2, a second container 3, and a tube 40. The first container 20 is closed at one end and has a container opening 22 at the other end. The first instant 30 passes through the container opening 22 and is received in the first container 2''. The second container 30 has a receiving portion 32 and a body portion. The end portion of the receiving portion 32 is folded toward the closed end to form a conical structure. The figure has two openings 36 opposite to the wall surface of the body portion 34 shown in the second figure. The second container 3 further has a retaining member 38 that covers the body portion 34. The tube member 40 is received in the second container 3 through the fixing member 38, and the tube member 38 is maintained in an upright state by the fixing member 38, and one end of the tube member 40 extends into the second container 3'' Located at the receiving portion 32, the other end of the tubular member 40 is openably engaged with a cover member 42. With the above structural and technical features, a sample pretreatment method using the liquid phase extraction apparatus 10 will be further described below. First, sample phase D, organic phase enthalpy, and phase enthalpy solution are separately prepared according to the analytes to be extracted, wherein the sample phase D, the organic phase enthalpy, and the receiving phase enthalpy solution are according to different samples, analytes, and experimental leaves. There are different configuration methods for the needs. As shown in the third figure, after the preparation of the above solution is completed, the sample phase D and a stirrer 24 are first added to the first 201231136 vessel 20, and then the organic phase 0 is added. The tube member 40 is then placed in the second container 40 through the fixing member 38, and the receiving phase is added to the second container 3, and the receiving phase A is contained in the receiving portion 32' to receive the phase. The liquid level of A is lower than the opening 36. As shown in the fourth figure, the tube member 40 and the second container 30 containing the receiving phase A are then placed in the first container 20, and the second container 30 is placed to the opening 36 below the organic phase. The liquid level is such that the organic phase flows into the second container 30 through the opening 36 to be in contact with the receiving phase A. After touching, the lid member 42 is covered and stirred for a period of time for extraction. After the extraction is completed, as shown in the fifth figure, the cover member 42 can be opened, and the receiving phase A can be taken out from the tube member 40. After the receiving phase a is taken out and after neutralization, the neutralized receiving phase A can be input into the detecting instrument for analysis. Accordingly, the liquid phase extraction device provided by the present invention and the sample pretreatment method using the liquid phase extraction device can simultaneously perform extraction and back extraction, and the liquid phase device has a simple structure and is easy to manufacture, and can be repeated after washing. It saves unnecessary supplies and costs, and is also very convenient and simple to operate. In another aspect, the liquid phase extraction apparatus provided by the present invention holds the receiving phase and protects it in the second container, and the sample phase cannot be combined with the receiving phase even if the organic phase solution is volatilized due to the extraction time or is dissolved in the water. ^ Pass, therefore, has better stability during the extraction process, and can also increase the selectivity of the organic solvent in the organic phase. The invention is further illustrated by an experimental example below. <Experimental Example> fbenzene (toluene methy U) is a kind of aromatic ring. 201231136 Hydrogen compound, widely used in industrial organic solvents, colorless, slightly sweet and irritating flammable liquid. The main source of terpene in nature is the production of toluene in crude oil fuels and gum trees, mainly through the petrochemical industry manufacturing process. However, toluene is often used in chemical products such as paints, glues and thinners, in the manufacture of coatings, in coating thinners, in adhesives and in rubber, as well as in printing and leather tanning processes. Toluene is fat-soluble and will soon enter the central nervous system via blood. It produces hallucinations and euphoria after smoking, and is extremely sensitive to external stimuli. It is easy to be impulsive and produces behavioral deviations. If excessive intake increases with blood concentration, it will cause mental disorder, movement disorder, and non-directional sense. In humans, acute oral benzene poisoning can cause a feeling of oropharyngeal burning, stomach pain, abdominal pain, hemoptysis, nausea, vomiting, and feelings of weakness. Other possible symptoms include dizziness, euphoria, chest tightness, and faltering. More serious include blurred vision, tremors, shallow and rapid breathing, paralysis, unconsciousness, and paralysis. Most of the symptoms are manifestations of central nervous system toxicity, so monitoring the amount of toluene exposure is important. In China's relevant research report, biodetection is the measurement of harmful substances and their metabolites in biological samples. This detected substance is called biomarker because it can make up for the lack of environmental detection. Many studies have been used in recent years. Biomarkers require many intermediate steps from the exposure of chemicals to the development of diseases, and there are many biological indicators that can be used to assess human exposure and damage. Generally, the biological indicators are divided into three categories, namely exp暴露sure bi〇marke r, health effect biomarker and susceptibility biomarker, which can be further subdivided into 5 categories. , intrinsic dose, biological effective dos e, early response, structural or functional change (a ered structure/function) and susceptibility biomarkers; intrinsic dose and bioavailable dose Classified as exposed biological indicators, early effects and structural or functional changes are indicators of health effects, each with different physiological significance. In the literature on toluene metabolism, hippuric acid and para-cresol are the main metabolites of human amino acids, which are easily exposed to dietary and other organic solvents, affecting the metabolism of toluene in humans. Therefore, it affects the concentration of para-cresol and hippuric acid in the urine, so hippuric acid and para-p-phenol are less suitable as low-concentration toluene exposure indicators. Therefore, the urinary metabolite ortho-cresol and benzylmercapt uric acid (BMA) are preferred indicators of exposure to benzene. Preparation of the extraction vertical column The extraction device in this experiment was assembled using 0.4 ml and 1.5 ml centrifuge tubes. The general centrifuge tube is made of polypropylene (p〇lypr〇pylene), because this experiment involves a more intense acid test solution. Therefore, a pre-lubricated tube/siliconized tube is used. Cut a suitable opening on both sides of the 1.5 ml centrifuge tube and drill a hole in the tube cap. Cut the bottom end of the 0.4 ml centrifuge tube and insert it into a 1.5 ml centrifuge tube cap, then clean it with ultrasonic waves. The device is shaken for 30 minutes, placed in a few minutes and then dried for use. Sample Preparation Collect urine from healthy adults who have not been exposed to stupid solvents, and centrifuge the urine for 13 minutes at 3000g for 5 minutes. Remove the supernatant for use. The precision scale is adjacent to each other; (para-cresol 99%, purchased from Alfa Aesar) l Omg is placed in a l〇ml measuring bottle, diluted with deionized water to the mark, and evenly mixed to obtain a concentration of lOOpp/ml. An ortho-cresol solution. The above procedure was repeated to obtain a solution of ortho-cresol 99% (purchased from Alfa Aesai) and a solution of benzylmercapturic aci d 99% (purchased from Alfa Aesar) at a concentration of 1000 pg/ml. Diluting and mixing o-position cresol, para-cresol and benzyl mercaptan acid at a concentration of 1000 pg/ml into a standard mixed solution of respective concentrations of 250 pg/ml, respectively, and taking 40 μβ to add a urine sample to i〇m The samples were added as a mixing standard of respective concentrations of lpg/ml. The extract is extracted from the sample vial (10 ml) and about 6.0 ml of the sample phase and the magnetic stirrer are added. Then 1.5 ml of the organic phase is added to the vial, wherein the sample phase has the experimental design requirements. Different formulations, the organic phase was prepared by mixing toluene (t〇luene, purchased from Merck 'Germany) and diethyl ether (dithyl ether, purchased from Merck, Germany) in a one-to-one volume. 〇4ml, 〇1M sodium hydroxide solution was added to the previously prepared centrifuge tube assembly as a receiving sample, and the sample tube containing the sample phase was placed in the mixing chamber, and stirred in a 50 C environment for about 15 minutes for extraction. After the extraction, the receiving phase is taken out by the injection needle and then analyzed by the detector H. The parameters and parameters are set in the analysis of the liquid chromatography tandem mass spectrometer. The liquid chromatograph is partially used in the liquid chromatography system (Waters ACQUITY Ultraper 201231136 formance LC system, purchased from the United States), separation The column is connected in series with pel i nomenex Kintex Cl8 (3.5 μιη '2.1x30 mm) and Supelico Diphenyl (2. :6 μιη, 2.1 x 150 mm), and at a column temperature of 4 (rc) at a flow rate of 〇j 5 ml/min. For equi-gradient elution, the mobile phase consisted of water and acetonitrile (acet〇ni • 1 1 , GR, purchased from Merck, Germany) in a volume ratio of 60 to 40. Mass spectrometer (Waters TQD mass spectrometer, purchased from UK) 4 injuries in negative ion mode (negative i〇n mode), in full-scan mode, with a mass range of 80~35〇m/z for detection. First change the capillary exit end After the voltage, the energy of the collision cell is changed, and the intensity of the broken ion signal intensity of the compound to be analyzed at different energies can be obtained. And in the tune mode, the voltage at the outlet end of the capillary is sequentially changed after the 'child ion scanning mode ( Daugh The ters ion scan, Dau-S can) analyzes the precursor ions (precurs〇ri〇n, [m_h]·) and sequentially changes the energy value of the collision cell to obtain different ion signals of each ion, and we Select the progeny ion that can get the most signal as the collision to the month 1. The parameters of the multiple reaction monitoring mode are as follows:
Capillary(KV) Cone(V) CE(V) 母離子 > 子離子 3.0 30V 20 252>123 3.0 30V 10 107>107 3.0 30V 10 107>107 分析物 ----- 下基硫醇酸 對位甲酚 ----_ 鄰位甲酚 201231136 屁通後品萃取前後之公折 第六圖所示為尿液添加曱酚標準品溶液萃取前後與曱 酚標準品溶液之實驗結果比較分析圖，其中（1)圖之樣品相 為3.1ml之展液樣本添加lpg/ml之甲紛標準品溶液，再加上 3.1ml、0.2M磷酸水溶液，並且該樣品相未經過萃取之分析數 據；（2)圖之樣品相為3.1ml水溶液添加lpg/mi曱盼標準品溶 液，再加入3.1m卜0.2M磷酸水溶液之分析數據；（3)圖之樣品 相為3.1ml尿液樣本添加.lpg/ml之甲酴標準品溶液，再加入 3.1m卜0.2M磷酸水溶液，並且該樣品相經過萃取之分析數據。 透過比較分析可以發現萃取前與萃取後，鄰位甲酚之訊號約有 7倍的差異；經過萃取後之鄰位甲酚訊號與甲酚標準品溶液之 鄰位甲酚訊號相比約有3.5倍的差距。 第七圖所示為尿液樣本添加节基硫醇酸標準品溶液萃 取前後與苄基硫醇酸標準品溶液之實驗結果比較分析圖， 其中（1)圖之樣品相為3.1ml尿液樣本添加lpg/ml之苄基硫 醇酸標準品溶液’再加入3.1ml、0.2M填酸水溶液，並且未經 過卒取之分析數據，（2)圖之樣品相為3.1ml水溶液添加lpg/ ml苄基硫醇酸標準品溶液’在加入3 lml、〇.2m磷酸水溶液 之分析數據；（3)圖之樣本相為3.1ml尿液添加1μδ/ιη1之苄基 硫醇酸標準品溶液，再加入3.1ml、〇.2]V[磷酸水溶液，並且該 樣本相經過萃取之分析數據。透過比較分析可以發現萃取前與 萃取後，鄰苄基硫醇酸之訊號約有將近1〇〇〇倍的差異；經過 萃取後之鄰苄基硫醇酸訊號與苄基硫醇酸標準品溶液之鄰苄 201231136 基硫醇酸訊號相比，則約有2倍的差距，顯見本發明對尿液樣 品之淨化對分析物訊號之影響甚矩。 ' 管柱德注I入測詖 管柱後注入測試分析主要是在管柱後注入鄰位甲酚、 蛉位甲酚以及节基硫醇酸稀釋並且混合成之標準混合溶 液，而在質譜儀上造成一連續之分析物訊號，接著在管柱 前方利用注人尿雜本萃取前、萃取後及無添加尿液樣本Capillary (KV) Cone (V) CE (V) parent ion > daughter ion 3.0 30V 20 252>123 3.0 30V 10 107>107 3.0 30V 10 107>107 Analyte----- Subunit thiol acid alignment Cresol----_ ortho-cresol 201231136 Folding before and after the product extraction, the sixth figure shows the comparative analysis of the experimental results of the urine and the indole standard solution before and after the extraction of the standard solution. The sample phase of (1) is a 3.1 ml sample of the liquid sample added with a standard solution of lpg/ml, plus 3.1 ml of a 0.2 M aqueous phosphoric acid solution, and the sample phase is not subjected to extraction analysis data; The sample phase of the sample is added to the 3.1 ml aqueous solution of lpg/mi 标准 standard solution, and then the analysis data of 3.1 m of 0.2 M phosphoric acid aqueous solution; (3) the sample phase of the sample is added to the 3.1 ml urine sample. lpg/ml The standard solution of the formazan was further added with a 3.1 m aqueous solution of 0.2 M phosphoric acid, and the sample phase was subjected to extraction analysis data. Through comparative analysis, it can be found that the signal of ortho-cresol is about 7 times different before and after extraction; the extracted ortho-cresol signal is about 3.5 compared with the ortho-cresol signal of the cresol standard solution. Double the gap. The seventh figure shows the comparison of the experimental results of the solution of the benzyl thiol acid standard solution before and after the extraction of the thiol acid standard solution in the urine sample. The sample phase of the sample (1) is 3.1 ml of the urine sample. Add lpg/ml of benzyl thiol acid standard solution' and add 3.1ml, 0.2M aqueous solution of acid, and did not pass the analysis data of stroke. (2) The sample phase of the sample is 3.1ml aqueous solution added lpg / ml benzyl Base thiol acid standard solution 'in the analysis data of adding 3 lml, 〇.2m phosphoric acid aqueous solution; (3) sample phase is 3.1ml urine added 1μδ / ιη1 benzyl thiol acid standard solution, and then added 3.1 ml, 〇.2] V [aqueous phosphoric acid, and the sample phase was subjected to extraction analysis data. Through comparative analysis, it can be found that the signal of o-benzyl thioalic acid is approximately 1〇〇〇 times different before and after extraction; the extracted o-benzyl thiol acid signal and benzyl thiol acid standard solution after extraction Compared with the benzyl 201231136-based thiol acid signal, there is about a difference of about 2 times. It is obvious that the purification of the urine sample of the present invention has a great influence on the analyte signal. After the pipe column is injected into the test column, the injection test is mainly carried out by injecting ortho-cresol, m-cresol and thiol acid in the column after the column is diluted and mixed into a standard mixed solution. A continuous analyte signal is generated, followed by pre-extraction, post-extraction, and no urine samples in the front of the column.
之鹽類溶液’以比較觀察萃取前後，基#效應對於分析物 離子訊號的影響。 實驗結果之分析比較如第八圖所示，⑴圖之樣品相為 3ml尿液樣本添加3mi、〇 2M磷酸 經過萃取後之分析數據；_之樣品相為6m丨甲酸^= 溶液之分析數據，圖之樣品相為3ml尿液樣本添加3m 卜〇通磷酸水溶液，並絲經過萃取之分析數據。透過 分析比較可以得知，尿液樣本經由萃取過後之情形，與甲 酸鹽類溶賴轉子訊號的抑麟_似，其麵分鐘左 右的訊號抑雜可能即為㈣魏所造成，但麵過半八 鐘後即無抑龍象。但是未經過萃取的尿液樣本造成分二 物離子訊號抑制相當日·，即使義六分鐘時 留時間時也有明顯抑制的現象。 / 結論 本發明所提供之液相萃取裝置，可同時進 卒取、減少液鮮取猶在有機溶_的 大幅降低尿樣樣品在質譜分析上的基質效應。此二 17 201231136 作與材料的取得皆取自實驗室常見的離心管與樣品瓶，故 成本相當低廉，除可重複使用外，更可便於同時進行大量 的樣品前處理。利用此液相萃取裝置來分析尿液樣品中甲 笨暴露的指標化合物鄰位曱酚與苄基硫醇酸，經實驗結果 證明此液相萃取裝置與萃取方法為一個十分簡便的前處理 方法’對於尿液樣品可同時達到淨化及濃縮之效果。 在此必須說明者為，以上配合圖式所為之詳細描述， 僅係為了說明本發明之技術内容及特徵而提供之一實施方 式，凡在本發明領域中具有一般通常知識之人，在暸解本籲 發明之技術内容及特徵之後，於不違背本發明之精神下， 所為之種種簡單之修飾、替換或構件之減省，皆應屬於以 下所揭示之申請專利範圍之内。 201231136 【圖式簡單說明】 第一圖為本發明一較佳實施例之立體圖； 第二圖為本發明一較佳實施例之剖視圖； ，第二圖為本發明一較佳實施例中，第一容器加入磁性 ㈣子、樣品相以及有機相，第二容器加入接收相並置入 管件’且第—容器尚未置人第—容器内之示意圖； ，第四圖為本發明一較佳實施例中，第一容器加入磁性 鲁 子、樣品相以及有機相，第二容器加入接收相並置入 e件，且第二容器置入第一容器内之示意圖； 一第五圖為本發明一較佳實施例中，從管件取出接收相 之示意圖； 第六圖為尿液樣本添加甲酚標準品溶液萃取前後與甲 酚標準品溶液之比較分析圖，其中標號⑴為尿液樣本添加 。kg ml之曱齡;^準品溶液，並且未經過萃取之實驗圖譜；標 號(2)為水;容液添加4_之甲紛標準品溶液之實驗圖譜· ^ ♦ 號⑶為尿液樣本添加Wml之甲紛標準品溶液，並且經過 取之實驗圖譜； /第七圖為尿液樣本添加节基硫醇酸標準品溶液萃取前 =與¥基硫_鮮品騎之峨讀圖，其巾標號⑴為 “液樣本添加1μβ/ιη1之节基硫醇酸標準品溶液，並且未經過 取之實驗H ^號(2)為水溶液添加丨^^之？基硫醇酸 標準品溶液之實驗圖譜;標號⑶為尿液樣本添加㈣W 基硫醇酸標準品溶液’並且經過萃取之實驗圖譜；以及 苐八圖為管柱後注入測試之比較分析圖，其中標號⑴ 201231136 為尿液樣本添加0.2M磷酸水溶液，並且經過萃取之實驗 圖譜；標號(2)為甲酸鹽類水溶液之實驗圖譜；標號(3)為尿 液樣本添加0.2M磷酸水溶液，並且未經過萃取之實驗圖 201231136 【主要元件符號說明】The salt solution was used to compare the effects of the base # effect on the analyte ion signal before and after the extraction. The analysis and comparison of the experimental results are shown in the eighth figure. (1) The sample phase is the analysis data of 3m urine sample added with 3mi and 〇2M phosphoric acid after extraction; the sample phase is the analysis data of 6m strontium formate ^= solution, The sample phase of the figure was added to a 3 ml urine sample by adding 3 m aqueous solution of dithizone phosphate, and the wire was subjected to extraction analysis data. Through analysis and comparison, it can be known that after the extraction of the urine sample, it is similar to the inhibition of the formic acid-like rotor signal. The signal suppression around the minute may be caused by (4) Wei, but over half of the surface. There is no dragon in the back of the bell. However, the unextracted urine sample caused a considerable suppression of the ion signal, which was significantly inhibited even when the time was left for six minutes. / Conclusion The liquid phase extraction device provided by the invention can simultaneously advance and reduce the liquid effect of the liquid sample, and greatly reduce the matrix effect of the urine sample on the mass spectrometry. These two 17 201231136 materials and materials are obtained from the centrifuge tubes and sample bottles commonly used in the laboratory, so the cost is quite low, in addition to reusable, it is easier to carry out a large number of sample pretreatment at the same time. The liquid phase extraction device was used to analyze the ortho-indolol and benzyl mercaptan acid in the urine sample, and the liquid phase extraction device and the extraction method were proved to be a very simple pretreatment method. For urine samples, the effect of purification and concentration can be achieved at the same time. It is to be understood that the foregoing detailed description of the drawings is merely illustrative of the technical aspects and features of the present invention, and those of ordinary skill in the field of the invention </ RTI> </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of a preferred embodiment of the present invention; FIG. 2 is a cross-sectional view of a preferred embodiment of the present invention; A container is provided with a magnetic (four) sub-sample, a sample phase and an organic phase, a second container is added to the receiving phase and placed in the tube member 'and the first container is not placed in the first container. FIG. 4 is a preferred embodiment of the present invention. Wherein the first container is added to the magnetic lining, the sample phase and the organic phase, the second container is added to the receiving phase and placed in the e-piece, and the second container is placed in the first container; In the preferred embodiment, the schematic diagram of the receiving phase is taken from the tube; the sixth figure is a comparative analysis of the cresol standard solution before and after the extraction of the phenol sample solution, wherein the label (1) is added to the urine sample. Kg ml of age; ^ standard solution, and unextracted experimental map; label (2) is water; liquid solution added 4_ of the standard solution of the standard solution · ^ ♦ No. (3) for the urine sample Wml's standard solution is taken, and the experimental map is taken; /The seventh picture is the addition of the thiol acid standard solution to the urine sample before the extraction = with the base sulphur _ fresh product riding the map, its towel The label (1) is the experimental sample of adding a 1 μβ/ιη1 thiol acid standard solution to the liquid sample, and adding the experimental H ^ number (2) to the aqueous solution to add the thiol acid standard solution. ; (3) Add the (IV) W-based thiol acid standard solution for the urine sample and extract the experimental map; and Figure 8 is the comparative analysis of the post-injection test, where the label (1) 201231136 adds 0.2M to the urine sample. An aqueous solution of phosphoric acid and an experimental chart of extraction; label (2) is an experimental map of a formate brine; label (3) is a 0.2 M aqueous solution of phosphoric acid for urine samples, and an unextracted experimental diagram 201231136 [Major component symbol description 】
液相萃取裝置10 第一容器20 容器口 22 攪拌子24 第二容器30 盛置部32 身部34 開口 36 固定件38 管件40 蓋件42 樣品相D 有機相〇 接收相ALiquid phase extraction device 10 First container 20 Container port 22 Stirrer 24 Second container 30 Surrounding portion 32 Body 34 Opening 36 Fixing member 38 Tube member 40 Cover member 42 Sample phase D Organic phase 接收 Receiving phase A
21twenty one