CN110025980B - Efficient concentrated solution-liquid micro-extraction device and method capable of realizing seamless combined chromatographic analysis - Google Patents
Efficient concentrated solution-liquid micro-extraction device and method capable of realizing seamless combined chromatographic analysis Download PDFInfo
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Abstract
The invention discloses a high-efficiency concentrated solution-liquid micro-extraction device and a method capable of realizing seamless combined chromatographic analysis.A trace extraction liquid collecting bottle is arranged at the upper end or the lower end, and is connected through an extraction enricher with an opening converging gradually, so that a sample can be automatically positioned at the upper layer or the lower layer after extraction layering, the collection effect of the extraction liquid containing high-concentration components can be greatly improved through further enrichment of the extraction enricher, the concentration and enrichment times of the sample are greatly improved under the condition that the extraction rate is not changed greatly, and the detection limit and the measurement limit of sample measurement are greatly reduced; the method achieves the purpose that the same instrument can be used for measuring trace substances, and greatly improves the measuring range of the instrument, thereby completing the work of high-grade instruments by using low-grade cheap instruments and measuring trace substances by using high-grade instruments.
Description
Technical Field
The invention belongs to the technical field of chemical analysis, and relates to a high-efficiency concentrated solution-liquid microextraction device and method capable of realizing seamless combined chromatographic analysis.
Background
The liquid-liquid extraction method is also called solvent extraction or extraction, and refers to the separation and extraction of components in a liquid mixture by using a solvent; liquid-liquid extraction is one of the most commonly used methods in the pretreatment of analytical test samples. The liquid-liquid extraction process includes adding selected solvent immiscible with liquid mixture, and utilizing the different solubility of its components in solvent to reach the separation or extraction aim.
In gas chromatography, the sample should be treated to an organic solvent, not to an aqueous solution; therefore, gas chromatography analysis of test liquid samples is often necessary with organic solvent extraction. The development trends of sample pretreatment are the minimization of solvent, the minimization of pollution, the continuity of operation and the zero-gap connection of sample treatment and analysis instruments. However, liquid-liquid extraction is a process in which two immiscible solvents compete for the component to be measured, so that at least one organic solvent must be involved, and the pollution and potential danger caused by the use of the organic solvent are difficult to avoid and can only be reduced as much as possible. The less organic solvent used, the less pollution and the less risk of burning and explosion, until the environmental pollution is minimized. However, at present, the national standard, the international standard and the domestic standard detect pollutants such as phenols, nitrobenzene and the like in a water sample, and during quantitative analysis, the sample amount is hundreds to one thousand milliliters, while the theoretical concentration multiple is only dozens of times. As specified in determination of aqueous nitrobenzene compounds by liquid-liquid extraction/solid phase extraction-gas chromatography (HJ 648-2013): 200ml of water sample is measured and added with 10.00ml of toluene for extraction, the theoretical concentration multiple is only 20 times, and the extraction system is huge, the organic solvent consumption is large, the pollution is large, and the organic solvent has hidden dangers of combustion, explosion and the like.
The extraction effect and enrichment degree of the liquid-liquid extraction method can directly influence the analysis and detection results. For some extraction, the enrichment degree is low, and the concentration of the extracted sample cannot reach the measurement limit of the instrument, so that the measurement work cannot be carried out. For some gas chromatographs, when the split sampling is not carried out, the response value of the lowest concentration of a standard curve specified by the national standard is only about 3 times of the noise, which barely reaches the detection limit and does not reach the measurement limit at all.
Disclosure of Invention
The invention solves the technical problem of providing a high-efficiency concentrated solution-liquid micro-extraction device and a method capable of realizing seamless combined chromatographic analysis, which can be used for on-line measurement by a chromatographic automatic sample injector without transferring after high-efficiency concentrated solution-liquid extraction of a sample, and overcomes the defects of large dosage of an extracting agent, serious pollution and waste, low extraction enrichment degree, processed samples even not reaching the measurement limit of an instrument and difficult continuous operation existing in the conventional liquid-liquid extraction.
The invention is realized by the following technical scheme:
a high-efficiency concentrated liquid-liquid micro-extraction device capable of realizing seamless combined chromatographic analysis comprises an uplink liquid-liquid micro-extraction device and a downlink liquid-liquid micro-extraction device, wherein the solvent density of the uplink liquid-liquid micro-extraction device is less than that of a sample to be detected, and the solvent density of the downlink liquid-liquid micro-extraction device is greater than that of the sample to be detected;
the ascending liquid-liquid micro-extraction device comprises a big tripe extraction bottle and an ascending micro extraction liquid collection bottle; the bottle diameter of the big belly extraction bottle gradually converges from the bottom to the bottle mouth at the top end, and the side wall at the bottom end is provided with a solvent supplement port;
the size of the ascending micro-extraction liquid collecting bottle is required to be placed in a sample tray of the automatic chromatographic sample injector, a funnel-shaped collector is arranged in the bottle, the bottom of the funnel-shaped collector is arranged at the bottom of the bottle, and the top of the funnel-shaped collector is connected with the bottle wall at the upper end; the bottom of the ascending micro extraction liquid collecting bottle is matched with the top bottle mouth of the big belly extraction bottle and can extend into the big belly extraction bottle to be connected with the sealed ground mouth, and the top bottle mouth is provided with a flexible bottle cap; the ascending micro extraction liquid collecting bottle is also provided with an inverted T-shaped bottom bottle stopper which can extend into and block the funnel-shaped collector;
the descending liquid-liquid micro-extraction device comprises an extraction bottle and a descending micro-extraction liquid collection bottle; the upper end of the extraction bottle is provided with a bottle opening, and the bottom of the lower bottle is communicated with a connecting pipe; the connecting pipe is a bell mouth or a columnar pipe with an upward big end, and the lower port of the connecting pipe is provided with a chromatographic sample bottle screw cap;
the size of the descending trace extraction liquid collecting bottle can be required to be placed in a sample plate of the automatic chromatographic sample injector, a bottle opening at the upper end of the descending trace extraction liquid collecting bottle is provided with a screw and can accommodate a lower opening of a connecting pipe, and the bottom of the descending trace extraction liquid collecting bottle is a sealed flat bottom; the upper bottle mouth is provided with a flexible bottle cap.
The batch extraction bottle tray is configured for the ascending micro extraction liquid collecting bottle, a plurality of pocket-shaped fixing cavities are arranged on a tray body of the batch extraction bottle tray, cushion pads are arranged in the pocket-shaped fixing cavities, and reinforcing covers are further arranged on the pocket-shaped fixing cavities; the bottom of the extraction bottle tray is provided with a sucker which can be fixed on the shaking table.
The device also comprises a double-layer extraction bottle tray configured for a descending trace extraction liquid collecting bottle, wherein the outer layer of the double-layer extraction bottle tray is in a stainless steel disc shape and can be fixed on a shaking table, and the inner layer of the double-layer extraction bottle tray is a foam buffer layer; a plurality of placing cavities capable of containing descending trace extraction liquid collecting bottles are formed in the foam buffer layer.
The bottle bottom of the ascending trace extract collecting bottle and the bottle mouth at the top end of the big belly extraction bottle are both provided with frosted surfaces;
the solvent supplementing port is made into a spiral port same as the chromatographic sample bottle;
and a flexible bottle cap of the ascending micro-extraction liquid collecting bottle is internally provided with a bottle pad, and the flexible bottle cap and the bottle pad allow a chromatographic syringe to penetrate through the spacer without liquid leakage.
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis, which is used for liquid-liquid micro-extraction when the solvent density is less than that of a sample to be detected, comprises the following operations:
1) accurately adding a certain volume of sample to be detected into the big belly extraction bottle, and accurately measuring 50-500 ul of extractant to inject into the big belly extraction bottle, wherein the liquid level of the sample to be detected is 2-4 ml away from the bottle mouth;
2) sealing the big belly extraction bottle after adding the sample and the extractant, and fully shaking for extraction; after extraction is balanced, placing and layering;
3) inserting a ground port at the lower end of a trace extraction liquid collecting bottle into a bottle port at the upper end of a big belly extraction bottle, and opening a spiral port cover at the upper end of the trace extraction liquid collecting bottle;
4) absorbing saturated saline as lifting liquid, slowly injecting the lifting liquid into a big belly extraction bottle from a solvent supplementing port to enable the liquid level in the bottle to rise until all organic layers containing samples to be detected enter a micro extraction liquid collecting bottle from a funnel-shaped collector, and taking down the collecting bottle after a screw cap with a screw opening at the upper end is covered; plugging an opening below the micro extraction liquid collecting bottle by using an inverted T-shaped bottom plug to finish primary extraction;
5) the collection vial is placed in a chromatographic autosampler sample tray to select chromatographic conditions to initiate chromatographic testing.
When the batch sample is detected, the operation of fully shaking and extracting is as follows: and (3) fixing the big belly extraction bottles of all the samples on a tray, placing the tray on a shaking table, continuously shaking for 20 minutes, then taking down the big belly extraction bottles, and layering the bottles.
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis, which is used for liquid-liquid micro-extraction when the solvent density is greater than that of a sample to be detected, comprises the following operations:
1) a spacer with a hole in the middle is arranged at the bottom end of the connecting extension pipe of the extraction bottle in a cushioning manner, and then a descending trace extraction liquid collecting bottle is tightly connected with the connecting extension pipe and sealed by a bottle cap; accurately adding a certain amount of sample to be detected from the upper end of the extraction bottle, and accurately measuring 50-500 ul of extracting agent with a certain volume and injecting into the extraction bottle;
2) fully shaking for extraction, standing for layering after extraction is balanced, and shaking the extraction bottle to allow the extractant to fully sink in a descending trace extraction liquid collecting bottle;
3) tightly covering the cover at the upper end of the extraction bottle, opening a connecting cover between the connecting extension pipe and the descending trace extraction liquid collecting bottle, and taking down the descending trace extraction liquid collecting bottle;
4) and after extraction is finished, placing the collection bottle in a sample tray of the chromatographic automatic sample injector, and performing chromatographic detection after chromatographic conditions are selected.
When the batch sample is detected, the operation of fully shaking and extracting is as follows:
firstly, fixing the automatic extraction trays of a batch of samples on a shaking table, fixing the extraction bottles of all the samples in an accommodating cavity, placing the fixed extraction bottles on a shaking table for continuously shaking for 20 minutes, then taking down the extraction bottles and layering the extraction bottles.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention uses the principle of liquid-liquid distribution, and selects the solvent which is not soluble with the sample solution but can well dissolve the substance to be detected as the micro-extraction agent. By arranging the micro extraction liquid collecting bottle at the upper end or the lower end, the micro extraction bottle is connected through an extraction enricher (a funnel-shaped collector or an extension tube) with an opening which is gradually converged, and a sample can be automatically positioned at the upper layer or the lower layer after extraction layering, so that the collection effect of the extraction liquid containing high-concentration components can be greatly improved by further enriching through the extraction enricher, the concentration enrichment multiple of the sample is greatly improved under the condition that the extraction rate is not changed greatly, and the detection limit and the measurement limit of sample measurement are greatly reduced; the method achieves the purpose that the same instrument can be used for measuring trace substances, and greatly improves the measuring range of the instrument, thereby completing the work of high-grade instruments by using low-grade cheap instruments and measuring trace substances by using high-grade instruments.
Furthermore, the extraction and the chromatogram are connected in a seamless manner, and after the sample is transferred to the ascending micro-extraction bottle or the descending micro-extraction bottle through the extraction enricher, the micro-extraction bottle can be directly placed into the chromatogram sample tray, and the injector can be pricked into the bottle cap for sampling, so that the operation is convenient, and the pollution is avoided; the batch oscillation extraction and the batch sample introduction can be realized through the arrangement of the batch extraction device, so that the complexity of the detection operation is greatly simplified; the extraction device and the sample prepared by the method can be used for gas chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, flow injection analysis, microcell ultraviolet-visible spectrum and fluorescence spectrum analysis, and can realize automatic sample injection determination of the chromatography analysis.
The invention reduces the whole extraction system due to the micro-extraction agent, so the environmental pollution is greatly reduced; the potential danger of organic solvent combustion explosion is greatly reduced; the testing cost is greatly reduced; the sampling cost is also greatly reduced; the test sensitivity is greatly improved; the extraction process is mechanized and automated, and the sample treatment conditions are uniform, so that the repeatability and accuracy of the test result are greatly improved.
Drawings
FIG. 1 is a schematic diagram of a liquid-liquid microextraction extraction flask with a solvent density less than that of a sample to be tested;
FIG. 2 is a schematic diagram of a liquid-liquid micro-extraction liquid collection bottle with a solvent density less than that of a sample to be tested;
FIG. 3 is a drawing of a liquid-liquid microextraction extraction flask with solvent density greater than that of the sample to be tested;
FIG. 4 is a collection bottle for liquid-liquid microextraction extract with solvent density greater than that of the sample to be tested;
FIG. 5 is a liquid-liquid batch micro-extraction gas chromatography tray with solvent density less than that of the sample to be tested;
FIG. 6 is a liquid-liquid bulk micro-extraction flask tray with solvent density greater than that of the sample to be tested.
Fig. 7 is a graph of the response behavior of different amounts of extractant for each of the substances, annotated: ai in the figure is the peak area of the object to be measured; a. thebIs the peak area of the side-normal n-heptane.
Detailed Description
The present invention will now be described in further detail with reference to the attached drawings, which are illustrative, but not limiting, of the present invention.
The invention designs a whole set of liquid-liquid micro-extraction device and an operation method, which can realize solvent micro-quantification, minimize environmental pollution and maximize the concentration ratio of a substance to be detected. The invention also designs a series of devices used for implementing the method, so that the method can realize automatic shaking extraction, extraction liquid does not need to be transferred, extraction phases are collected in micro-extraction sample bottles after being layered, and then the sample bottles are directly placed in a sample tray of the automatic chromatograph sample feeder for automatic sample feeding and determination.
Referring to fig. 1-7, a high-efficiency concentrated liquid-liquid microextraction device capable of seamless joint chromatographic analysis comprises an uplink liquid-liquid microextraction device with a solvent density smaller than that of a sample to be detected and a downlink liquid-liquid microextraction device with a solvent density larger than that of the sample to be detected;
the ascending liquid-liquid micro-extraction device comprises a big tripe extraction bottle 1 and an ascending micro extraction liquid collection bottle 4; the bottle diameter of the big belly extraction bottle 1 gradually converges from the bottom to the top bottle mouth 3, and the side wall of the bottom end is provided with a solvent supplement port 2;
the size of the ascending micro-extraction liquid collecting bottle 4 can be required to be placed in a sample tray of the automatic chromatographic sample injector, a funnel-shaped collector 5 is arranged in the bottle, the bottom of the funnel-shaped collector 5 is arranged at the bottom of the bottle, and the top of the funnel-shaped collector is connected with the bottle wall at the upper end; the bottom of the ascending micro extraction liquid collecting bottle 4 is matched with the top bottle mouth 3 of the big belly extraction bottle 1 and can extend into the ground mouth to be connected, and the top bottle mouth is provided with a flexible bottle cap; the ascending micro extraction liquid collecting bottle 4 is also provided with an inverted T-shaped bottom bottle stopper 12 which can extend into and block the funnel-shaped collector 5;
the descending liquid-liquid micro-extraction device comprises an extraction bottle 6 and a descending micro-extraction liquid collection bottle 9; the upper end of the extraction bottle is provided with a bottle opening, the diameter of the bottle is converged downwards from the middle lower part of the bottle, and the bottom of the lower end of the bottle is communicated with a connecting pipe 7; the connecting pipe 7 is a horn mouth or a columnar pipe with an upward big end, and the outer side of the lower end of the connecting pipe 7 is provided with a screw and is provided with a screw cap;
the descending trace extraction liquid collecting bottle 9 can be placed in a sample plate of the automatic chromatographic sample injector according to the size requirement, the bottle opening at the upper end of the descending trace extraction liquid collecting bottle is provided with a screw opening and can accommodate the lower opening of the extension tube 7, and the bottle bottom is a sealed flat bottom; the upper mouth is provided with a flexible cap 10.
The batch extraction bottle tray is configured for the ascending micro extraction liquid collecting bottle 4, a plurality of pocket-shaped fixing cavities 10 are arranged on a tray body of the batch extraction bottle tray, cushion pads are arranged in the pocket-shaped fixing cavities, and reinforcing covers are further arranged on the pocket-shaped fixing cavities; the bottom of the extraction bottle tray is provided with a sucker which can be fixed on the shaking table.
The device also comprises a double-layer extraction bottle tray configured for the descending trace extraction liquid collecting bottle 9, the outer layer of the double-layer extraction bottle tray is in a stainless steel disc shape and can be fixed on the shaking table, and the inner layer is a foam buffer layer; a plurality of placing cavities capable of containing descending trace extraction liquid collecting bottles are formed in the foam buffer layer.
The bottle bottom of the ascending trace extract collecting bottle 4 and the bottle mouth at the top end of the big belly extraction bottle 1 are both provided with frosted surfaces;
the solvent supplementing port 2 is made into a spiral port same as the chromatographic sample bottle;
and a bottle pad is also arranged in the flexible bottle cap of the ascending micro-extraction liquid collecting bottle 4, and the flexible bottle cap and the bottle pad allow a chromatographic syringe to penetrate through the spacer without liquid leakage.
The ratio of the diameter of the opening at the upper end of the connecting pipe 7 to the diameter of the opening at the bottom of the extraction bottle 6 is 1: 5-20.
The extraction method is given below.
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis, which is used for liquid-liquid micro-extraction when the solvent density is less than that of a sample to be detected, comprises the following operations:
1) accurately adding a certain volume of sample to be detected into the big belly extraction bottle, and accurately measuring 50-500 ul of extractant to inject into the big belly extraction bottle, wherein the liquid level of the sample to be detected is 2-4 ml away from the bottle mouth;
2) sealing the big belly extraction bottle after adding the sample and the extractant, and fully shaking for extraction; after extraction is balanced, placing and layering;
3) inserting a ground port at the lower end of a trace extraction liquid collecting bottle into a bottle port at the upper end of a big belly extraction bottle, and opening a screw port at the upper end of the trace extraction liquid collecting bottle;
4) absorbing saturated saline as lifting liquid, slowly injecting the lifting liquid into a big belly extraction bottle from a solvent supplementing port to enable the liquid level in the bottle to rise until all organic layers containing samples to be detected enter a micro extraction liquid collecting bottle from a funnel-shaped collector, and taking down the collecting bottle after a screw cap with a screw opening at the upper end is covered; plugging an opening below the micro extraction liquid collecting bottle by using an inverted T-shaped bottom plug to finish primary extraction;
5) the collection vial is placed in a chromatographic autosampler sample tray to select chromatographic conditions to initiate chromatographic testing.
When the batch sample is detected, the operation of fully shaking and extracting is as follows: and (3) fixing the big belly extraction bottles of all the samples on a tray, placing the tray on a shaking table, continuously shaking for 20 minutes, then taking down the big belly extraction bottles, and layering the bottles.
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis, which is used for liquid-liquid micro-extraction when the solvent density is greater than that of a sample to be detected, comprises the following operations:
1) a spacer with a hole in the middle is arranged at the bottom end of an extension tube of the extraction bottle in a cushioning manner, and then a descending trace extraction liquid collecting bottle is tightly connected with the extension tube and sealed by a connecting cover with threaded openings at both ends; accurately adding a certain amount of sample to be detected from the upper end of the extraction bottle, and accurately measuring 50-500 ul of extracting agent with a certain volume and injecting into the extraction bottle;
2) fully shaking for extraction, standing for layering after extraction is balanced, and shaking the extraction bottle to allow the extractant to fully sink in a descending trace extraction liquid collecting bottle;
3) tightly covering the upper end of the extraction bottle, opening a connecting cover between the extension pipe and the descending trace extraction liquid collecting bottle, and taking down the descending trace extraction liquid collecting bottle;
4) and after extraction is finished, placing the collection bottle in a sample tray of the chromatographic automatic sample injector, and performing chromatographic detection after chromatographic conditions are selected.
When the batch samples are detected, the operation of fully shaking and extracting is as follows:
firstly, fixing the automatic extraction trays of a batch of samples on a shaking table, fixing the extraction bottles of all the samples in an accommodating cavity, placing the fixed extraction bottles on a shaking table for continuously shaking for 20 minutes, then taking down the extraction bottles and layering the extraction bottles.
Specific examples are given below.
The invention provides a whole set of liquid-liquid micro-extraction device and operation, which comprises:
the solvent density is less than the liquid-liquid micro-extraction device of the sample to be measured and the solvent density is greater than the liquid-liquid micro-extraction device of the sample to be measured;
1) the liquid-liquid micro-extraction device with the solvent density smaller than that of the sample to be detected comprises an extraction bottle and a trace extraction liquid collecting bottle; the main body of the extraction bottle is similar to a pycnometer, and the upper bottle mouth of the extraction bottle is tightly matched with the lower ground mouth of the micro extraction liquid collecting bottle; the lower end of the extraction bottle main body is provided with a solvent replenishing port which is made into a spiral port same as the universal chromatographic sample bottle;
the micro-extraction liquid collecting bottle can be placed in a sample plate of the automatic chromatographic sample injector, the outer part of the lower port of the micro-extraction liquid collecting bottle can be tightly matched with the main body of the extraction bottle through fine grinding, so that liquid leakage cannot occur, and the bottom end is not sealed; the upper port is provided with a screw port, and the bottle cap and the rubber spacer are required to ensure that the chromatographic syringe can freely penetrate through the spacer without leakage;
2) the liquid-liquid micro-extraction device with the solvent density larger than that of the sample to be detected comprises an extraction bottle and a trace extraction liquid collecting bottle;
the extraction bottle is characterized in that a 2 cm-saving glass tube is connected to the lower end of the headspace bottle, a chromatographic sample bottle screw cap is sleeved on the glass tube in advance, and the lower end opening of the glass tube is fired into a bell mouth and ground flat;
the micro extraction liquid collecting bottle can be placed in a sample plate of the automatic chromatographic sample injector; the lower end of the trace extraction liquid collecting bottle is a sealed flat bottom, the upper end opening is provided with a screw, and the bottle cap and the rubber spacer are required to ensure that the chromatographic syringe can be freely pricked through the spacer but cannot leak liquid.
3) Batch extraction bottle tray
Liquid-liquid batch micro-extraction bottle tray with solvent density less than that of the sample to be detected: the extraction bottle tray is a plastic tray body, 25-36 pocket-shaped devices are arranged on the extraction bottle tray, and the extraction bottle tray is fixed on a special shaking table for extraction after being covered.
The solvent density is larger than the tray of the liquid-liquid micro-extraction bottle of the sample to be detected: the outer layer is in a stainless steel disc shape and is fixed on the shaking table; the inner layer is a cube made of foam and is slightly lower than the outer layer. The inner layer has 25-36 holes, and the bottom of the holes has a hole with a diameter of 1.3cm, into which an extract collecting bottle can be inserted.
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis is characterized by comprising the following operations:
1) liquid-liquid microextraction with solvent density less than that of sample to be tested
(1) Manual extraction of the sample: accurately adding a certain volume of sample to be detected into the extraction flask, reserving 2-4 ml of liquid surface space according to the size of the extraction flask, and accurately measuring 50-500 ul of extracting agent with a certain volume and injecting into the extraction flask.
(2) After the sample and the extractant are added, a polytetrafluoroethylene grinding port cover is used for covering an extraction bottle, and the extraction time is slightly longer than that of the traditional extraction because the extractant has small volume and small contact area with the water phase by sufficient shaking extraction); after the extraction is balanced, the layers are placed (if necessary with slight ultrasound) and the teflon cover is removed slightly to make the layers clear.
(3) And replacing the polytetrafluoroethylene cover with a trace extraction liquid collecting bottle, enabling a ground port at the lower end of the collecting bottle to be matched with a ground port at the upper end of the extraction bottle, and opening a spiral port of the collecting bottle.
(4) The saturated saline salt is absorbed by a syringe with proper volume to have salting-out function, so that the extraction efficiency can be improved, and different effects of extracting various salts are different, so that the salt is determined by specific extraction). Slowly injecting the organic solvent into the extraction bottle from the solvent supplementing port to enable the liquid level in the bottle to rise until all the organic layers enter the collection bottle, covering the spiral cover, and taking down the collection bottle (because the cover is sealed, the extractant cannot flow out under the action of atmospheric pressure);
in order to ensure that the extractant does not flow out during subsequent sample injection, a special inverted T-shaped polytetrafluoroethylene plug is used for tightly plugging an opening below a collecting bottle, so that one extraction is completed, and then the next extraction is sequentially carried out.
5) After the whole extraction is completed, the collection bottle is placed in a sample tray of a chromatographic automatic sampler for selecting chromatographic conditions for determination.
6) Automatic extraction of batch samples: and (3) operating according to the step 1) and the step 1), when shaking extraction is needed, putting the whole extraction flask on the extraction flask tray for fixing, continuously shaking on a shaking table for 20 minutes, taking down the extraction flask, and layering, wherein the subsequent operation is the same as the steps (3) to (4) in the step 1).
A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis comprises the following operations:
liquid-liquid microextraction with solvent density greater than that of sample to be tested
(1) Manual extraction of individual samples: punching a through hole with the aperture of 2-2.3 mm in the center of a spacer of a chromatographic sample bottle by using a puncher, filling the through hole at the joint of a spiral cover and an extractant collecting bottle to ensure that the solution is not leaked, connecting a screw port with an extraction main body bottle and the collecting bottle, opening a cover at the upper end of the extraction bottle, accurately adding a certain amount of sample to be detected, accurately measuring 50-500 ul of extractant with a certain volume, and injecting the extractant into the bottle;
(2) fully shaking for extraction (because the volume of the extractant is small and the contact area with a water phase is small, the extraction time is longer than that of the traditional extraction), placing for layering after the extraction is balanced, and shaking the extraction bottle to ensure that the extractant fully sinks in a small collection bottle;
(3) tightly covering the cover at the upper end of the extraction bottle, slightly opening the connecting screw of the extraction bottle and the collecting bottle, taking down the collecting bottle, replacing a new cover and a spacer, screwing, placing to be tested, and continuously processing the next sample;
(4) after the whole extraction is finished, placing the collection bottle in a sample tray of a chromatographic automatic sample injector, and selecting chromatographic conditions for determination;
(5) automatic extraction of batch samples: firstly fixing automatic extraction discs of a batch of samples on a shaking bed body, then operating according to the steps 1) (1), covering an extraction bottle added with the samples and an extractant, then placing the extraction bottle in a fixed sleeve, and automatically extracting for 20 minutes on the shaking bed;
adjusting the angle of the fixing sleeve to enable the extraction flask to be vertical to the horizontal plane, standing for layering (slightly knocking the wall of the flask if necessary) to enable the extractant to be collected in a collection small flask, and performing the following operations like (3) to (4) in the step 2).
Specific detection examples are given below, aiming at comparing sensitivity, peak area amplification, signal response values, and assay stability in the case of high sensitivity with the new method.
The benzene series is one of main pollutants in water, and is taken as a measuring object in the early national standard, and benzene, toluene, three xylene isomers and styrene are mainly measured. The invention is issued for improving the enrichment degree of extraction, solving the gapless butt joint of sample treatment and chromatographic test and reducing environmental pollution.
As a performance test study, it is not necessary to test all substances, for which purpose C in the benzene series is selected6C7C8Each of which is as follows: benzene, toluene, styrene as molecular probes were examined for process performance and compared to the conventional process.
1. Material
A self-made liquid-liquid extraction bottle kit (comprising a specially-made extraction liquid collecting small bottle, a polytetrafluoroethylene lower opening plug of the collecting small bottle and a polytetrafluoroethylene upper opening plug of the extraction bottle); micro syringes of 100ul, 250ul and 500ul, medical syringes of 1.0ml and 5.0 ml; a shaking table special for extraction is made; a gas chromatograph equipped with an FID detector; 2L volumetric flask; 8L reagent bottles; 250ml of pear-shaped separating funnel; laboratory conventional glassware was several.
N-hexane (chromatographically pure, benzene-free, n-heptane); benzene, toluene, styrene (analytically pure); blue oil soluble dyes, or other oil soluble dyes whose color is readily observable.
2. Preparation of samples
2.1 extractant a n-hexane solution containing side mark 1ul/1ml and blue layer separating indicator 0.5 mg/ml. The chromatographic analysis has an internal standard method, i.e. quantitative and stable substances which can be separated from all peaks are respectively added into a standard sample and a sample for testing, so as to achieve the satisfactory testing purpose. There is also an external standard method, i.e. a method of testing after making a working curve with a standard sample.
In order to verify the extraction effect, correct the chromatographic sample introduction amount deviation and the experimental deviation caused by the volatilization and concentration of a small amount of extractant in the subsequent operation after extraction balance, a reagent is added into the extractant in advance, the performance of the reagent is required to be easily soluble in the extractant and insoluble in water, chromatographic peaks can be completely separated from all peaks, the reagent is added into the extractant, the reagent not only replaces an internal standard but also can play other deviation rectifying performances, and in order to distinguish from the internal standard and an external standard, the reagent is called a side standard in the experiment, and the side standard reagent in the experiment is n-heptane.
Extracting agent b is n-hexane solution containing 0.2ul/1ml blue layered indicator 0.5mg/ml side mark
2.2 mixing original three benzene series by 500ul micro syringe accurately take benzene, toluene, styrene each 500ul in 2.0ml sample bottle shake.
2.3 preparation of simulated water sample (1.00 ul/L.), adding water to the scale in 2000ml volumetric flask, accurately sucking 18.00ul of mixed benzene original sample in 2.2 with 25ul micro syringe, injecting into the flask, shaking fully until the mixed benzene is completely dissolved in water, transferring the solution to 8L reagent bottle, measuring distilled water twice with proper 2000ml volumetric flask, adding to 8L reagent bottle, and shaking up. The concentrations of benzene, toluene and styrene in the solution were all 1.00 ul/L. Preparation of a simulated water sample (0.200 ul/L.). The simulated water samples (1.00 ul/L.) were diluted 5-fold.
3. Extracting and collecting
Taking 8 clean extraction bottles, opening the bottle caps, sucking 3.0ml of water out of a water sample (about 103ml) filled with 2.3 water samples by using a 5ml syringe, adding a certain amount of extractant, covering a polytetrafluoroethylene cover, manually or semi-automatically extracting for 20min to ensure extraction balance, standing for layering, removing the polytetrafluoroethylene cover, replacing an extract collection bottle, unscrewing a spiral cover of the collection bottle, fully sucking the water by using the 5ml syringe, slowly injecting the water from the lower port of the extraction bottle to ensure that all organic phases in the extraction bottle enter the collection bottle, injecting excessive water with the amount of less than 200ul when the amount of the extractant is small (less than 200ul), removing the syringe, tightly covering the spiral cover of the collection bottle to ensure air tightness, slightly taking down the collection bottle, observing the liquid level, injecting air slowly by using the 1ml syringe when a small amount of water exists at the lower end of the collection bottle, penetrating through a rubber spacer at the upper end of the collection bottle to ensure that all the water is pushed out, and covering the opening at the lower end of the collecting bottle by using an inverted T-shaped polytetrafluoroethylene plug, and placing the collecting bottle on a gas chromatography automatic sample introduction disc for automatic determination. In the experiment, Agilent 7890 with two instruments is used for sample injection, and GC9800(N/TF) with the same creation is used for sample injection manually without an autosampler.
4. Analytical testing
4.1 measurement of chromatographic Peak area response behavior
Testing was performed by gas chromatography: gas chromatograph 1.GC7890 agilent company usa (this instrument is used only for spot check of results, main test is performed on GC 9800N/TF); GC9800N/TF Shanghai was co-inventively equipped with FID detectors. Chromatographic conditions are as follows: chromatographic column DB-5MS, 30m × 0.25mm × 0.5um, column temperature 80 deg.C, injection port temperature 200 deg.C, and detector temperature 200 deg.C. The pressure of carrier gas high-purity nitrogen is 0.22Mpa, the pressure of hydrogen is 0.03Mpa, the pressure of air is 0.03Mpa, and the split-flow and sample injection split ratio is 10: 1. Under the condition, 1.0ul of sample is injected for chromatographic test, and the test results are recorded in the following table, and the variation trend is shown in the following figure 7.
TABLE 1 response behavior of different amounts of extractant for extraction of various substances
As can be seen from FIG. 7, the peak area of the measured substance rapidly increases with the decrease in the volume of the extractant, Ai/AbThe value is approximately multiplied with the multiplied reduction of the volume of the extractant, and when the extractant is 0.10ml (100ul), the concentration of each benzene series is approximately concentrated by 1000 times, namely the theoretical concentration is 1ul/ml, the concentration is approximately 1.0ul/ml of the side target, and A of the benzene series is Ai/AbThe value swings around 1, in line with theoretical predictions. However, not exactly equal to 1, due to the different chromatographic response values of the substances, the different distribution coefficients, and the tendency of the side marks to distribute to water. The response trend of benzene is not perfect as that of toluene and styrene, because the benzene peak is close to the solvent peak, and the trace impurities in the solvent have influence on the benzene peak, and the optimal condition can be preferably solved. It can be predicted that organic substances with high boiling points in water, which are difficult to measure by headspace gas chromatography, are very suitable for this method.
4.2 testing of extraction reproducibility
In consideration of the ease of operation of the experiment and the like, 0.2ul/L of the water sample containing the probe was extracted with 200ul (0.20ml, side-normal heptane concentration 0.20ul/ml) of the extractant in 8 extraction flasks and subjected to a reproducibility test. Extracting to obtain A by the extraction method in the patenti/AbValue, statistically calculated, of benzene ABenzene and its derivatives/AbRSD of ═ 2.1; toluene AToluene/AbRSD of ═ 1.4; styrene (meth) acrylic acid esterAStyrene (meth) acrylic acid ester/AbThe RSD of the method is 1.7 which is superior to that of the RSD of the liquid-gas chromatography reported by the national standard GB11890-89 of 4.1-11.9, and the RSD of the carbon disulfide extraction gas chromatography of 3.5-9.9.
The embodiments given above are examples of implementing the present invention, and the present invention is not limited to the above examples. Any non-essential addition and replacement made by the technical characteristics of the technical scheme of the invention by a person skilled in the art belong to the protection scope of the invention.
Claims (9)
1. A high-efficiency concentrated liquid-liquid micro-extraction device capable of realizing seamless combined chromatographic analysis is characterized by comprising an uplink liquid-liquid micro-extraction device and a downlink liquid-liquid micro-extraction device, wherein the solvent density of the uplink liquid-liquid micro-extraction device is less than that of a sample to be detected, and the solvent density of the downlink liquid-liquid micro-extraction device is greater than that of the sample to be detected;
the ascending liquid-liquid micro-extraction device comprises a big tripe extraction bottle (1) and an ascending micro extraction liquid collection bottle (4); the bottle diameter of the big belly extraction bottle (1) gradually converges from the bottom to the top bottle mouth (3), and the side wall of the bottom end of the big belly extraction bottle is provided with an ejection liquid input port (2);
the size of the ascending micro-extraction liquid collecting bottle (4) is required to be placed in a sample tray of the automatic chromatographic sample injector, a funnel-shaped collector (5) is arranged in the bottle, the bottom of the funnel-shaped collector (5) is opened, and the top of the funnel-shaped collector is connected with the bottle wall at the upper end; the bottom of the ascending micro extraction liquid collecting bottle (4) is matched with the top end bottle mouth (3) of the big belly extraction bottle (1), can extend into the bottle and is connected with the bottle mouth in a grinding sealing way, and the bottle mouth at the top of the collecting bottle is provided with a flexible bottle cap; the ascending trace extraction liquid collecting bottle (4) is also provided with an inverted T-shaped bottom bottle stopper, the bottom bottle stopper can extend into and block the lower opening of the funnel-shaped collector (5), and the collecting bottle (4) can be freely placed in a chromatographic sample tray after plugging;
the descending liquid-liquid micro-extraction device comprises an extraction bottle (6) and a descending micro-extraction liquid collection bottle (9); the upper end of the extraction bottle is provided with a bottle opening, the diameter of the bottle is converged downwards from the middle lower part of the bottle, and the bottom of the lower bottle is communicated with a connecting pipe (7); the connecting pipe (7) is a bell mouth or a columnar pipe with an upward big end, the lower end of the connecting pipe (7) is also in a bell mouth shape, and a spiral cap is sleeved outside the connecting pipe;
the descending trace extraction liquid collecting bottle (9) can be placed in a sample plate of the automatic chromatographic sample injector according to the size requirement, the bottle opening at the upper end of the descending trace extraction liquid collecting bottle is provided with a screw opening and can accommodate the lower end opening of the connecting pipe (7), and the bottle bottom is a sealed flat bottom; the upper end bottle mouth is provided with a flexible bottle cap (10).
2. The high-efficiency concentrated liquid-liquid microextraction device capable of realizing seamless joint chromatographic analysis according to claim 1, further comprising a batch extraction bottle tray configured for the ascending micro-extraction liquid collecting bottle (4), wherein a plurality of pocket-shaped fixed cavities (10) are arranged on a tray body of the batch extraction bottle tray, cushions are arranged in the pocket-shaped fixed cavities, and reinforcing covers are further arranged on the pocket-shaped fixed cavities; the bottom of the extraction bottle tray is provided with a sucker which can be fixed on the shaking table.
3. The high-efficiency concentrated liquid-liquid microextraction device capable of realizing seamless joint chromatographic analysis according to claim 1, further comprising a double-layer extraction bottle tray configured for descending trace extraction liquid collection bottles (9), wherein the outer layer is in the shape of a stainless steel disc capable of being fixed on a shaking table, and the inner layer is a foam buffer layer; a plurality of placing cavities capable of containing descending trace extraction liquid collecting bottles are formed in the foam buffer layer.
4. The high-efficiency concentrated solution-liquid micro-extraction device capable of realizing seamless joint chromatographic analysis according to claim 1, wherein the bottle bottom of the ascending micro-extraction liquid collecting bottle (4) and the bottle mouth at the top end of the big tripe extraction bottle (1) are provided with frosted surfaces;
the ejection liquid input port (2) is made into a spiral port which is the same as the chromatographic sample bottle;
and a bottle pad is also arranged in the flexible bottle cap of the ascending micro-extraction liquid collecting bottle (4), and the flexible bottle cap and the bottle pad allow a chromatographic syringe to penetrate through the spacer without liquid leakage.
5. The high-efficiency concentrated liquid-liquid microextraction device capable of realizing seamless joint chromatographic analysis according to claim 1, wherein the ratio of the diameter of the opening at the upper end of the connecting pipe (7) to the diameter of the opening at the bottom of the extraction bottle (6) is 1: 5-20.
6. A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis is characterized in that liquid-liquid micro-extraction when the density of a solvent is smaller than that of a sample to be detected comprises the following operations:
(1) accurately adding a certain volume of sample to be detected into the big belly extraction bottle, and accurately measuring 50-500 ul of extractant to inject into the big belly extraction bottle, wherein the liquid level of the sample to be detected is 2-4 ml away from the bottle mouth;
(2) sealing the big belly extraction bottle after adding the sample and the extractant, and fully shaking for extraction; after extraction is balanced, placing and layering;
(3) inserting a ground port at the lower end of a trace extraction liquid collecting bottle into a bottle port at the upper end of a big belly extraction bottle, and opening a screw port at the upper end of the trace extraction liquid collecting bottle;
(4) absorbing saturated saline water as lifting liquid, slowly injecting the lifting liquid into a big belly extraction bottle from an ejection liquid input port to enable the liquid level in the bottle to rise until all organic layers containing samples to be detected enter a micro extraction liquid collecting bottle from a funnel-shaped collector and slightly more liquid is collected, and taking down the collecting bottle after a screw cap of an upper end screw port is covered; plugging an opening below the micro extraction liquid collecting bottle by using an inverted T-shaped bottom plug to finish primary extraction;
(5) the collection vial is placed in a chromatographic autosampler sample tray to select chromatographic conditions to initiate chromatographic testing.
7. The method for high-efficiency concentrated liquid-liquid microextraction capable of seamless combined chromatography according to claim 6, wherein when detecting a batch of samples, the operation of sufficient shaking extraction is as follows: and (3) fixing the big belly extraction bottles of all the samples on a tray, placing the tray on a shaking table, continuously shaking for 20 minutes, then taking down the big belly extraction bottles, and layering the bottles.
8. A high-efficiency concentrated liquid-liquid micro-extraction method capable of realizing seamless combined chromatographic analysis is characterized in that liquid-liquid micro-extraction when the density of a solvent is greater than that of a sample to be detected comprises the following operations:
(1) a spacer with a hole in the middle is arranged at the bottom end of a connecting pipe of the extraction bottle in a cushioning manner, and then a descending trace extraction liquid collecting bottle is tightly connected with the connecting pipe and sealed by a connecting cover with threaded ports at both ends; accurately adding a certain amount of sample to be detected from the upper end of the extraction bottle, and accurately measuring 50-500 ul of extracting agent with a certain volume and injecting into the extraction bottle;
(2) fully shaking for extraction, standing for layering after extraction is balanced, and shaking the extraction bottle to allow the extractant to fully sink in a descending trace extraction liquid collecting bottle;
(3) tightly covering the cover at the upper end of the extraction bottle, opening a connecting cover between the connecting pipe and the descending trace extraction liquid collecting bottle, and taking down the descending trace extraction liquid collecting bottle;
(4) and after extraction is finished, placing the collection bottle in a sample tray of the chromatographic automatic sample injector, and performing chromatographic detection after chromatographic conditions are selected.
9. The method of claim 8, wherein the batch sample testing is performed by performing a full shake extraction process by:
firstly, fixing the automatic extraction trays of a batch of samples on a shaking table, fixing the extraction bottles of all the samples in an accommodating cavity, placing the fixed extraction bottles on a shaking table for continuously shaking for 20 minutes, then taking down the extraction bottles and layering the extraction bottles.
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| US4118313A (en) * | 1973-11-03 | 1978-10-03 | Dynamit Nobel Aktiengesellschaft | Process and apparatus for mass transfer between heterogeneous systems |
| US3988116A (en) * | 1975-02-27 | 1976-10-26 | The Dow Chemical Company | Apparatus for countercurrent liquid-liquid extraction |
| NO151900C (en) * | 1980-09-26 | 1985-06-26 | Elkem As | PROCEDURE AND APPARATUS FOR CONTINUOUS EXCLUSION OF SOLUBLE COMPONENTS FROM SOLIDS, PARTICULARLY BY THE EXCLUSION OF MINERALS |
| CN1004787B (en) * | 1986-11-25 | 1989-07-19 | 林梦生 | Pulsation type method of solid-liquid continuous reverse extraction |
| CN101279720A (en) * | 2007-12-25 | 2008-10-08 | 天津大学 | Extraction apparatus and method for producing hydrogen peroxide by anthraquinone method |
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