TWI400117B - A liquid phase extraction apparatus, and a sample pretreatment method using the liquid phase extraction apparatus - Google Patents

Description

Liquid phase extraction device and sample pretreatment method using the same liquid phase extraction device

The present invention relates to an extraction apparatus, and more particularly to a liquid phase extraction apparatus and a sample preparation method using the liquid phase extraction apparatus.

The pretreatment of the sample has the effects of reducing the matrix effect, increasing the analyte content and increasing 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. I284198 provides a rapid chemical detection. Pretreatment methods and equipment to reduce sample preparation time.

In general, sample pretreatment techniques are distinguished by solid phase extraction and liquid phase extraction, where solid phase extraction can reduce or avoid the use of organic solvents, but is more expensive and can result in many Waste; liquid phase extraction uses a relatively large amount of organic solvent, which is easy to cause environmental pollution, but it is easy 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, which uses the concept of partitioning in liquid-liquid extraction to place droplets of organic solvents that are not incompatible with water in a flowing In the large droplets, the technique of continuous extraction. (Liu, H., Dasgupta, PK (1996). Analytical chemistry in a drop. Solvent extraction in a microdrop. Analytical Chemistry, 68 (11), 1817-1821.)

In the same year, Jeannot and Cantwell published a new single-drop micro-extraction technique in which an organic solvent droplet was fixed with a Teflon rod, and the organic solvent droplet was immersed in a rotating water sample solution stirred by a magnet. After a period of extraction, Injected into a gas chromatograph for analysis. (Jeannot, MA, & Cantwell, FF (1996). Solvent microextraction into a single drop. Analytical Chemistry, 68 (13), 2236-2240.)

In 1997, Jeannot and Cantwell further improved the aforementioned single-drop solvent microextraction method, using a microinjected oblique tip to replace the Teflon rod to fix the organic solvent droplets, and immersing the organic solvent droplets in the aqueous phase sample for extraction, and It can be injected into the gas chromatograph more conveniently. (Jeannot, MA, & Cantwell, FF (1997). Mass transfer characteristics of solvent extraction into a single drop at the tip of a syringe needle. Analytical Chemistry, 69 (2), 235-239.)

In 1998, Ma and Cantwell first combined extraction and back extraction (extraction/back-extraction) in the same step. In the case where the analyte belongs to a basic molecule, the analyte is a neutral molecule at a pH of 13 in the donor phase, and the analyte is distributed to the organic phase; in the receiving 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., & Cantwell, FF (1999). Solvent microextraction with simultaneous back-extraction for sample cleanup and preconcentration: preconcentration into a single microdrop. Analytical Chemistry, 70 (18), 3912-3919.)

However, when Ma and Cantwell's extraction apparatus perform single-drop solvent microextraction, the droplets are difficult to maintain and are easily affected by bubbles and matrix microparticles in the sample, which may fall during the extraction process, thus failing to improve the extraction process. Conditions such as temperature and stirring rate.

In 1999, Pedersen-Bjergaard and Rasmussen published hollow fiber liquid-phase microextraction (HF-LPME), which uses a hollow fiber tube made of polyproplyene (PP) to protect the solvent. Disadvantages. (Pedersen-Bjergaard, S., & Rasmussen, KE (1999). Liquid-liquid-liquid microextraction for sample preparation of biological fluids prior to capillary electrophoresis. Analytical Chemistry, 71 (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 injected into the gas chromatograph for analysis, and the hollow fiber liquid phase was micro. The extraction 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 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. (Hou, L., & Lee, HK (2003). Dynamic three-phase microextraction as a sample preparation technique prior to capillary electrophoresis, Analytical Chemistry, 75 (11), 2784-2789.)

Compared with the single-drop microextraction method, when the extraction solvent volume of the two is the same, the hollow fiber tube can provide a large contact area between the organic solvent and the sample, and the solvent droplets are not easily lost due to the support of the hollow fiber tube support. In the water sample. In addition, the hollow fiber tube can be fixed to the microinjection needle tip, and the injection needle type push-pull injection needle push rod can be used to improve the extraction efficiency of the analyte. (Jiang, X., Oh, SY, & Lee, HK (2005). Dynamic liquid-liquid-liquid microextraction with automated movement of the acceptor phase. Analytical Chemistry, 77 (6), 1689-1695.)

In 2004, Jang and Lee developed a solvent bar 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 rod, and then the solvent rod was placed. The solution is rotated with the magnet to perform the distribution effect to achieve the extraction effect. (X. Jang, & HK Lee (2004). Solvent bar micro extraction. Analytical Chemistry, 76 (18), 5591-5596.)

Compared with single-drop microextraction and static hollow fiber tube liquid extraction, solvent rod microextraction method achieves higher extraction efficiency by increasing the interface contact area and agitation rate.

In 2004, Schellin, Hauser and Popp expanded the capacity of the hollow fiber tube. Because it can be loaded with a large volume of solvent and combined with a large volume injection of a gas chromatograph, it can effectively increase the detection limit of the analyte. (Schellin, 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. Journal of Chromatography A, 1040 ( 2), 251-258.)

In 2008, Pan et al. used a PCR centrifuge tube as a material for three-liquid phase extraction. The receiving phase was first placed in the front end of the PCR centrifuge tube, and then the organic phase was placed and covered in the receiving phase. Then, the PCR phase tube was filled with the sample phase. When it is full, it is inserted into a sample bottle filled with the sample phase, and placed in a three-phase extraction. (Pan, W., Xu, H., Cui, Y., Song, D., & Feng, YQ (2008). Improved liquid-liquid-liquid microextraction method and its application to analysis of four phenolic compounds in water samples. Journal of Chromatography A, 1203 (1), 7-12.)

In combination with the above various liquid phase extraction techniques, the single-drop solvent micro-extraction method has been developed to utilize hollow fiber tubes to support and protect the extraction solvent, and to increase the extraction surface area and solvent stability, because the droplets are not easily maintained. Since the pores on the surface of the hollow fiber tube are selective, the macromolecule can be blocked and the macromolecule cannot enter 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, hollow fiber tubes may also cause problems of easy contamination and hole blockage for complex matrix samples, making hollow fiber tubes generally incapable of being reused. In addition, the use of an organic phase that is too small in volume tends to cause problems of dissolution and loss of volatility, and the type of organic phase that can be applied is limited.

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, reusable, and sample pretreatment using the liquid phase extraction apparatus The method has a relatively simple operation step.

To achieve the foregoing objective, the present invention provides a liquid phase extraction apparatus comprising a first container, a second container, and a tube member, wherein the second container is housed in the first container, and the second container has a shelf And a body portion, the container portion can hold a solution, the body having an opening that communicates with the inside and outside of the second container; the tube member is received in the second container.

The present invention further provides a sample pretreatment method using the liquid phase extraction apparatus described above, comprising the steps of: (a) adding an organic phase to the first container containing the sample phase; (b) placing the tube in the second a container, and a receiving phase is added to the second container; (c) placing the tube member with the second container containing the receiving phase into the first container, such that the organic phase enters the second container through the opening; d) Remove the receiving phase from the tube.

Accordingly, the liquid phase extraction apparatus and the sample preparation method using the liquid phase extraction apparatus provided by the present invention have a simple structure and are easy to manufacture, and can be repeatedly used after being cleaned, and are also very stable and convenient in operation.

The following is a detailed description of the preferred embodiments of the invention and the accompanying drawings in which: FIG. A cross-sectional view of a preferred embodiment; in a preferred embodiment of the invention, a first container is provided with a magnetic stirrer, a sample phase and an organic phase, a second container is added to the receiving phase and a tubular member is placed, and the second container is not yet A schematic diagram of being placed in a first container; in a preferred embodiment of the invention, a first container is added with a magnetic stirrer, a sample phase and an organic phase, a second container is added to the receiving phase and a tubular member is placed, and the second A schematic view of the container being placed in the first container; the fifth figure is a schematic view of the receiving phase taken out from the pipe member in a preferred embodiment of the present invention; and the sixth figure is a standard for adding cresol standard solution to the urine sample before and after extraction with cresol standard Comparative analysis of the solution, wherein the label (1) is a 1 μg/ml cresol standard solution for the urine sample, and the experimental map is not extracted; the label (2) is a 1 μg/ml cresol standard added to the aqueous solution. The experimental chart of the liquid; the label (3) is a 1 μg/ml cresol standard solution for the urine sample, and the experimental chart is extracted; the seventh picture is the addition of the benzyl thiol acid standard solution to the urine sample before and after extraction. A comparative analysis diagram of a benzyl thiol acid standard solution, wherein the label (1) is a 1 μg/ml benzyl thiouric acid standard solution added to the urine sample, and the experimental map is not subjected to extraction; the label (2) is an aqueous solution. Add 1 μg/ml of the experimental map of the benzyl thiol acid standard solution; label (3) to add 1 μg / ml of benzyl thiol acid standard solution to the urine sample, and extract the experimental map; and the eighth figure A comparative analysis chart for the post-injection test, wherein the label (1) is a 0.2 M phosphoric acid aqueous solution for the urine sample, and the extraction experiment is performed. Map; label (2) is the experimental map of the formate brine; label (3) is the experimental sample of the urine sample added with 0.2M aqueous phosphoric acid and not extracted.

As shown in the first and second figures, a liquid phase extraction apparatus 10 according to a preferred embodiment of the present invention includes a first container 20, a second container 30, and a tube member 40.

The first container 20 is closed at one end and has a container opening 22 at the other end.

The second container 30 is received through the container opening 22 and received in the first container 20. The second container 30 has a receiving portion 32 and a body portion 34. One end of the receiving portion 32 is closed, and the inner diameter of the receiving portion 32 is gradually contracted toward the closed end to form a conical structure. As shown in the second figure, the wall surface of the body portion 34 is oppositely opened with two openings 36. The second container 30 further has a fixing member 38 that covers the body portion 34.

The tube member 40 is received in the second container 30 through the fixing member 38. The tube member 40 is maintained in an upright state by the fixing member 38, and one end of the tube member 40 extends into the second container 30. The cover portion 32 has a cover member 42 that is openably closed at the other end of the tubular member 40.

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 O, and receiving phase A solution are prepared according to the type of analyte to be extracted, wherein sample phase D, organic phase O, and receiving phase A solution are in accordance with different samples, analytes, and experimental design requirements. There are different configuration methods. As shown in the third figure, after the preparation of the above solution is completed, the sample phase D and a stir bar 24 are first added to the first vessel 20, and then the organic phase O is added.

The tube member 40 is then placed in the second container 40 through the fixing member 38, and the receiving phase A is added to the second container 30. At this time, the receiving phase A is contained in the holding portion 32, so that the receiving phase A is received. The liquid level is lower than the opening 36.

As shown in the fourth figure, the tube 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 O. The liquid level is such that the organic phase O flows into the second container 30 through the opening 36 to come into contact with the receiving phase A, and the lid member 42 is covered and stirred for a while 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 neutralized, 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 can reduce unnecessary consumables and costs, and it is 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 mutually received even if the organic phase solution is volatilized due to the extraction time or is dissolved in the water. Therefore, it 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>

Toluene or methylbenzene is an aromatic ring hydrocarbon widely used in industrial organic solvents, colorless, slightly sweet and irritating flammable liquid. The main source of toluene 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, and 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 central disorders such as mental confusion, movement disorders, and non-directional sense. In humans, acute oral toluene 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 cases include blurred vision, tremors, shallow and rapid breathing, paralysis, unconsciousness, and paralysis. Most of the symptoms are manifestations of central nervous system toxicity, so it is important to monitor the amount of toluene exposure.

In China's relevant research report, biodetection is to measure the harmful substances and their metabolites in biological samples. This detected substance is called biomarker, because it can make up for the deficiency of environmental detection. Many studies have been used in recent years. Biological indicators range from chemical exposure to disease production, and many intermediate steps are required. 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, exposure biomarker, health effect biomarker and susceptibility biomarker, which can be further subdivided into five categories, each being an intrinsic dose. Biological effective dose, early response, structural or functional changes and susceptibility to biological indicators; intrinsic doses and biologically effective doses are classified as exposed biological indicators, early effects and Structural or functional changes are indicators of health effects, each with different physiological implications.

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 urine, so hippuric acid and para-cresol are less suitable as low-concentration toluene exposure indicators. Therefore, the urinary metabolites ortho-cresol and benzylmercapturic acid (BMA) are preferred indicators of exposure to toluene.

Production of extraction equipment

The extraction device in this experiment was assembled using 0.4ml and 1.5ml centrifuge tubes. The general centrifuge tube is made of polypropylene. Because this experiment involves more intense acid-base solution, the sputum-treated centrifuge tube is used. (pre-lubricated tube/siliconized tube).

Cut a suitable opening in the tube wall on both sides of the 1.5ml centrifuge tube, and drill a hole in the tube cover, cut the bottom end of the 0.4ml centrifuge tube, insert it into the 1.5ml centrifuge tube cap, and then clean it by ultrasonic wave. The device is shaken for 30 minutes, placed in a few minutes and then dried for use.

Sample configuration

The urine of healthy adults who had not been exposed to the toluene solvent was collected, and the urine was centrifuged at 3000 g for five minutes, and then the supernatant was taken out for use.

Separate cresol (para-cresol 99%, purchased from Alfa Aesar) 10mg was placed in a 10ml liquid bottle, diluted with deionized water to the mark, and evenly mixed to obtain a concentration of 1000μg/ml. Phenol solution. The above procedure was repeated to obtain a solution of para-cresol (ortho-cresol 99%, purchased from Alfa Aesar) and benzylmercaptoic acid 99% (purchased from Alfa Aesar) at a concentration of 1000 μg/ml.

After diluting and mixing o-position cresol, para-cresol and benzyl mercaptan acid at a concentration of 1000 μg/ml into a standard mixed solution each having a concentration of 250 μg/ml, 40 μg of the urine sample was taken and added to 10 ml, and each became a respective one. Samples were added at a mixing standard of 1 μg/ml.

Combination and extraction of extraction devices

After placing about 6.0 ml of the sample phase and the magnetic stirrer in the sample vial (10 ml), 1.5 ml of the organic phase was added to the vial, wherein the sample phase was formulated according to the experimental design requirements, and the organic phase was Toluene (purchased from Merck, Germany) and diethyl ether (dithyl ether, purchased from Merck, Germany) were mixed in a one to one volume.

After adding 0.4 ml of 0.1 M sodium hydroxide solution as the receiving phase to the previously prepared centrifuge tube assembly, the centrifuge tube assembly was placed in a sample bottle containing the sample phase, and stirred at 50 ° C for about 150 minutes for extraction. After the extraction is completed, the receiving phase is taken out and neutralized by an injection needle, and then injected into a testing instrument for analysis.

Conditions and parameter settings of liquid chromatography tandem mass spectrometer

In the analysis of the liquid chromatograph tandem mass spectrometer, the liquid chromatograph uses a high-performance liquid chromatography system (Waters ACQUITY Ultraperformance LC system, purchased from the United States), and the separation column is Phenomenex Kintex C18 (3.5 μm, 2.1×30 mm) was connected in series with Supelco Diphenyl (2.6 μm, 2.1×150 mm), and isocratic extraction was carried out at a column temperature of 40 ° C at a flow rate of 0.15 ml/min. It was prepared by mixing water and acetonitrile (GR, purchased from Merck, Germany) in a volume ratio of 60 to 40.

The mass spectrometer (Waters TQD mass spectrometer, purchased from the UK) was partially detected in a negative ion mode and detected in a mass range of 80-350 m/z in a full-scan mode. After changing the voltage at the outlet end of the capillary and then changing the energy of the collision cell, the intensity of the broken ion signal at different energies of the compound to be analyzed can be obtained. And in the tune mode, after sequentially changing the voltage at the outlet end of the capillary, the precursor ion ([MH] ^- ) is selected in the daughter ion scan mode (Dau-Scan), and By changing the energy value of the collision cell, the ion signals of different ions of each ion can be obtained, and we choose to obtain the daughter ion with the largest signal as the energy of the collision cell used. The parameters of the multiple reaction monitoring mode are as follows:

Comparative analysis of urine samples before and after extraction

The sixth figure shows the comparison of the experimental results of the cresol standard solution before and after extraction with the cresol standard solution. The sample phase of the sample (1) is a sample of 3.1 ml of urine and 1 μg/ml of the sample is added. A phenol standard solution, plus 3.1 ml, 0.2 M aqueous phosphoric acid solution, and the sample phase is not subjected to extraction analysis data; (2) the sample phase is 3.1 ml aqueous solution added 1 μg / ml cresol standard solution, and then added Analytical data of 3.1ml, 0.2M phosphoric acid aqueous solution; (3) The sample phase is 3.1ml urine sample, adding 1μg/ml cresol standard solution, then adding 3.1ml, 0.2M phosphoric acid aqueous solution, and the sample phase passes Analytical data for extraction. 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 benzyl thiol acid standard solution before and after the extraction of the benzyl thiol acid standard solution in the urine sample. The sample phase of the sample (1) is 3.1 ml of the urine sample. Add 1 μg/ml of benzyl thiol acid standard solution, add 3.1 ml, 0.2 M aqueous phosphoric acid solution, and analyze the data without extraction; (2) add 1.3 μl/ml benzyl sulfide to the sample phase of the sample. Alkyd standard solution, the analysis data of adding 3.1ml, 0.2M phosphoric acid aqueous solution; (3) The sample phase of the sample is 3.1ml urine, adding 1μg/ml of benzyl thiol acid standard solution, and then adding 3.1ml, 0.2 M 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 mercaptan is approximately 1000 times different before and after extraction; the extracted o-benzyl mercaptan signal and the benzyl thiol acid standard solution Compared with the base 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.

Post-injection test

The post-column injection test analysis mainly involves injecting ortho-cresol, para-cresol and benzyl mercaptan acid into the standard mixed solution after the column, and causing a continuous analyte signal on the mass spectrometer. Then, in front of the column, the salt solution before, after and after the extraction of the urine sample was used to compare the effects of the matrix effect on the analyte ion signal before and after the extraction.

The analysis and comparison of the experimental results are as shown in the eighth figure. (1) The sample phase is 3 ml of urine sample, 3 ml, 0.2 M phosphoric acid aqueous solution is added, and the sample phase is analyzed after extraction; (2) the sample phase of the sample It is the analysis data of 6 ml of formate aqueous solution; (3) the sample phase is 3 ml of urine sample, 3 ml, 0.2 M aqueous phosphoric acid solution, and the analytical data without extraction. Through analysis and comparison, it can be known that the urine sample is similar to the inhibition of the ion signal by the formate solution after the extraction, and the signal suppression in about two minutes is probably caused by the formate, but after passing through There is no inhibition after half a minute. However, the unextracted urine sample caused significant inhibition of the analyte ion signal, and even after about six minutes, the retention time of cresol was significantly inhibited.

in conclusion

The liquid phase extraction device provided by the invention can simultaneously perform extraction and back extraction, reduce the limitation of the liquid solvent extraction technology on the types of organic solvents, and greatly reduce the matrix effect of the urine sample on mass spectrometry. The production and materials of this device are taken from the centrifuge tubes and sample bottles commonly used in the laboratory, so the cost is quite low. In addition to being reusable, it is convenient to carry out a large number of sample pretreatment at the same time. The liquid phase extraction device was used to analyze the ortho-cresol and the benzyl mercaptan acid in the urine sample. The experimental results show that the liquid phase extraction device and the extraction method are a very simple pretreatment method. Urine samples can simultaneously purify and concentrate.

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 who have ordinary knowledge in the field of the invention </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt;

10. . . Liquid phase extraction device

20. . . First container

twenty two. . . Container mouth

twenty four. . . Stirrer

30. . . Second container

32. . . Survival Department

34. . . Body

36. . . Opening

38. . . Fastener

40. . . Pipe fittings

42. . . Cover

D. . . Sample phase

O. . . The organic phase

A. . . Receiving phase

The first figure is a perspective view of a preferred embodiment of the present invention;

The second drawing is a cross-sectional view of a preferred embodiment of the present invention;

In a third embodiment of the present invention, a first container is provided with a magnetic stirrer, a sample phase, and an organic phase, a second container is added to the receiving phase and the tubular member is placed, and the second container is not yet placed in the first container. schematic diagram;

4 is a schematic view showing a first container in which a magnetic stirrer, a sample phase and an organic phase are added, a second container is added to a receiving phase and a tubular member is placed, and a second container is placed in the first container. ;

Figure 5 is a schematic view showing the receiving phase taken out from the pipe member in a preferred embodiment of the present invention;

The sixth figure is a comparative analysis of the cresol standard solution before and after extraction with the cresol standard solution. The label (1) is a 1 μg/ml cresol standard solution added to the urine sample and is not extracted. The experimental map; the label (2) is an experimental map of adding 1 μg/ml of the cresol standard solution to the aqueous solution; the label (3) is a urine sample to which 1 μg/ml of the cresol standard solution is added, and the extracted experimental map is obtained;

The seventh figure is a comparative analysis of the solution of benzyl thiol acid standard solution before and after extraction with benzyl thiol acid standard solution. The label (1) is 1 μg/ml of benzyl thiol added to the urine sample. An acid standard solution, and an unextracted experimental map; label (2) is an experimental map of a solution of 1 μg/ml of a benzyl mercaptan acid standard solution added to an aqueous solution; and (3) a 1 μg/ml benzyl group for a urine sample. Base thiol acid standard solution, and an experimental profile of the extraction;

The eighth figure is a comparative analysis chart of the post-injection test. The label (1) is a 0.2M phosphoric acid aqueous solution added to the urine sample, and the experimental map is extracted; the label (2) is the experimental map of the formate aqueous solution; (3) An experimental map in which a 0.2 M aqueous phosphoric acid solution was added to the urine sample and was not extracted.

10. . . Liquid phase extraction device

20. . . First container

30. . . Second container

36. . . Opening

40. . . Pipe fittings

Claims (10)

A liquid phase extraction device comprising: a first container; a second container received in the first container, and the second container has a receiving portion and a body, the receiving portion can hold the solution, the The body has an opening that communicates with the inside and outside of the second container; and a tubular member is received in the second container, and one end of the tube is located at the receiving portion. The liquid phase extraction device according to claim 1, wherein the second container has a fixing member, and the fixing member is disposed on the second container, and the tube member is disposed through the fixing member. The liquid phase extraction apparatus according to claim 1, wherein the inner diameter of the holding portion gradually contracts toward the closed end to have a conical structure. A sample pretreatment method using a liquid phase extraction apparatus according to claim 1, comprising the steps of: (a) adding an organic phase to the first container containing the sample phase; (b) placing the tube member The second container, and the receiving phase is added to the second container; (c) the tube is placed into the first container together with the second container containing the receiving phase, and the organic phase enters the second container via the opening And (d) remove the receiving phase from the tube. The sample pretreatment method according to claim 4 of the patent application further comprises a step (e) of preparing a sample phase solution. The sample pretreatment method according to claim 4, further comprising a step (f) of preparing an organic phase solution. The sample pretreatment method according to claim 4 of the patent application further comprises a step (g) of preparing a receiving phase solution. The sample pretreatment method according to claim 4, further comprising a step (h) of bringing the organic phase and the receiving phase into contact with each other for a period of time. The sample preparation method of claim 4, further comprising a step (i) of neutralizing the receiving phase taken out of the tube. The sample pretreatment method according to claim 9, further comprising a step (j) of inputting the neutralized receiving phase into the detecting instrument.