CN219111061U - High-flux trace biological sample extracting solution purifying device - Google Patents

Abstract

The utility model discloses a high-flux trace biological sample extracting solution purifying device, which relates to the technical field of clinical examination and life science, and has the technical scheme that: the liquid storage device comprises a pump, a plurality of liquid delivery pipelines and a plurality of liquid storage bottles, wherein filter elements are arranged in the liquid storage bottles, the end parts of the liquid delivery pipelines are connected with the filter elements, and the end parts of the liquid delivery pipelines, which are far away from the filter elements, penetrate through the liquid storage bottles and are connected with the inlet ends of the pump; the outlet end of the pump is provided with an infusion pipeline, the end, far away from the pump, of the infusion pipeline is provided with a connector, the outlet end of the connector is provided with a liquid separating device, the outlet end of the liquid separating device is provided with a filtering device, and the outlet end of the filtering device is provided with a liquid collecting device. The device and method utilize microliter level extraction and purification of the extraction solution at higher back pressure.

Description

High-flux trace biological sample extracting solution purifying device

Technical Field

The utility model relates to the technical field of clinical examination and life science, in particular to a high-flux trace biological sample extracting solution purifying device.

Background

According to the needs of research, the detection of biological samples is mainly aimed at measuring small molecular index substances, effective components, active components, macromolecular compositions and the like in the biological samples so as to display the characteristics of the biological environment and biological process represented by the biological samples, and the biological samples are used for physiological and biochemical research. After the conventional sample is obtained, the steps of homogenization, extraction, purification and detection are generally carried out, and the common purification comprises the modes of solid phase extraction, liquid-liquid extraction, solid-liquid extraction, physical phase separation and the like, but in clinical and mass life science research, the common extraction and purification mode has the requirement of large sample demand due to the limitation and the requirement of sample taking, so that the purification requirement of the micro sample is difficult to meet, and under the condition of low concentration of the detected target substance, the purification contradiction is particularly prominent, and the problems of inaccurate test value and large test error are caused. In response to these problems, methods of microextraction, such as solid-phase microextraction and solid-dispersion extraction, have been developed in recent years.

The Solid Phase Microextraction (SPME) technology is based on the fact that fused quartz fibers coated with a stationary phase are adopted to adsorb and enrich substances to be detected in a sample, and has the greatest characteristics that analytes can be concentrated while extraction is carried out, but a mainstream adsorption extraction mode can be seen, the problem that the sample is partially sampled again exists, in addition, the selectivity of a filler also influences the enrichment and the accuracy of effective substances of the sample, and the inaccuracy and the poor quantitative effect of a detection result are caused.

Meanwhile, the traditional SPME technology in the market at present can only process one sample at a time, and the use flux is limited; in addition, in the purification of the solid dispersion method commonly used in recent years, certain requirements exist on the sample quantity, the treatment of the trace liquid sample is limited, and the corresponding target is difficult to effectively recover or enrich in a uniform environment completely, so that the detection is inaccurate.

Disclosure of Invention

The utility model aims to provide a high-flux trace biological sample extracting solution purifying device and method, which utilize micro liter level extracting and purifying of extracting solution under higher back pressure.

The technical aim of the utility model is realized by the following technical scheme: the high-flux trace biological sample extracting solution purifying device comprises a pump, a plurality of infusion pipelines and a plurality of liquid storage bottles, wherein filter elements are arranged in the liquid storage bottles, the end parts of the infusion pipelines are connected with the filter elements, and the end parts of the infusion pipelines, which are far away from the filter elements, penetrate through the liquid storage bottles and are connected with the inlet ends of the pump; the outlet end of the pump is provided with an infusion pipeline, the end, far away from the pump, of the infusion pipeline is provided with a connector, the outlet end of the connector is provided with a liquid separating device, the outlet end of the liquid separating device is provided with a filtering device, and the outlet end of the filtering device is provided with a liquid collecting device;

the connector is internally provided with porous fibers and a stainless steel sieve plate, and the bottom of the porous fibers is fixedly connected with the top of the stainless steel sieve plate;

the liquid separating device comprises a liquid separating disc, a plurality of conical pipes and a first porous plate; the first porous plate is positioned in the liquid separation disc to divide the liquid separation disc into an upper part and a lower part, and the bottom of the first porous plate is fixedly connected with the tops of the conical tubes at the same time; the top of the liquid separation disc is connected with the bottom of the connector;

the filtering device comprises a filtering disc, a porous plate II and a plurality of micro filling columns; the porous plate is positioned in the filter disc to divide the filter disc into an upper part and a lower part, and a plurality of micro filling columns are respectively arranged on the through holes of the porous plate II in a penetrating way up and down; the tops of the micro filling columns are in one-to-one correspondence with the bottoms of the conical tubes and are detachably connected with the bottoms of the conical tubes;

the micro-filling column is filled with purifying filler.

Further: the liquid collecting device comprises a base and a plurality of liquid collecting bottles, and the liquid collecting bottles are all arranged on the base; the openings of the liquid collecting bottles correspond to the outlet ends of the micro filling columns respectively.

Further: the purifying filler is silica gel, C ₁₈ One of diatomite and polystyrene-divinylbenzene.

Further: sealing rings are sleeved at the bottoms of the conical pipes.

A method for purifying a high-flux trace biological sample extracting solution is characterized by comprising the following steps: the method specifically comprises the following steps:

s1: connecting the pump, the infusion pipeline, the connector and the liquid separating device, starting a high flow rate, removing bubbles, and closing the pump when the solution flowing out of the conical pipe is continuous fluid;

s2: connecting the filtering device to the liquid separating device, then starting the pump, activating the prefilled purified filler, and closing the pump;

s3: and (3) placing the extracting solution in the activated micro-filling column, connecting the filtering device with the liquid separating device again, starting the pump to perform elution, and collecting the eluent in the liquid collecting bottle.

In summary, the utility model has the following beneficial effects:

1. the purification device utilizes the elution purification mode of a sample under a higher pressure environment, so that on one hand, the selectivity of the sample to a target substance is increased, and on the other hand, the loss of the sample target substance is reduced;

2. the micro-filling column is filled with purified filler, so that an arch bridging effect formed by the filler with different particle sizes can be utilized, a pressure-resistant filling environment is formed under unidirectional fluid, the use of a sieve plate is abandoned, in addition, under higher pressure, the compressibility and polarity of the fluid can be changed, the material distribution relationship formed between the fluid and the solid filler can be changed, and the extraction efficiency of the material is improved;

3. the sample adopts a full-pass elution method, so that the transfer loss of the sample is effectively solved, the recovery rate of the target substance is improved, and the accuracy of subsequent detection is also improved.

Drawings

FIG. 1 is a schematic diagram of a purification apparatus for high throughput trace biological sample extractive solution in accordance with embodiment 1 of the present utility model;

fig. 2 is a schematic structural view of a connector in embodiment 1 of the present utility model;

FIG. 3 is a schematic view showing the structure of a liquid separating apparatus in embodiment 1 of the present utility model;

FIG. 4 is a schematic view showing the structure of a filtering apparatus according to embodiment 1 of the present utility model;

FIG. 5 is a schematic view showing the structure of a micro-packed column in example 1 of the present utility model;

fig. 6 is a schematic structural view of a liquid collecting device in embodiment 1 of the present utility model.

In the figure: 1. a filter element; 2. a liquid storage bottle; 3. a pump; 4. a connector; 41. a porous fiber; 42. stainless steel sieve plate; 5. a liquid separating device; 51. a solvent bin; 52. a first porous plate; 53. a conical tube; 54. a seal ring; 6. a filtering device; 61. a filter tray; 62. a micro-packed column; 63. a porous plate II; 64. purifying the filler; 7. a liquid collecting device; 71. a base; 72. a liquid collecting bottle; 8. an infusion line; 9. an infusion pipeline.

Detailed Description

The utility model is described in further detail below with reference to fig. 1-6.

Example 1: the high-flux trace biological sample extracting solution purifying device comprises a pump 3, 2 infusion pipelines 8 and 2 liquid storage bottles 2, wherein filter elements 1 are arranged in the 2 liquid storage bottles 2, and the filter elements 1 are connected with the inlet end of the pump 3 through the infusion pipelines 8; in this embodiment, the pump 3 is preferably a dual-pump 3 head high-pressure pump 3, the outlet end of the pump 3 is connected with the connector 4 through an infusion pipeline 9, in this embodiment, the infusion pipeline 8 and the infusion pipeline 9 are preferably peek pipes or stainless steel pipes with narrow inner diameters (165-360 μm), the outlet end of the connector 4 is fixedly connected with the liquid separating device 5, the outlet end of the liquid separating device 5 is detachably connected with the inlet end position of the filtering device 6, the liquid collecting device 7 is positioned at the outlet end position of the filtering device 6, and the liquid collecting port corresponds to the outlet of the filtering device 6;

the connector 4 is internally provided with a porous fiber 41 and a stainless steel sieve plate 42, and as shown in fig. 2, the bottom of the porous fiber 41 is fixedly connected with the top of the stainless steel sieve plate 42; the stainless steel sieve plate 42 has a pore diameter of 0.22 μm, maintains back pressure with the infusion pipeline 9, realizes effective liquid separation, and eliminates bubbles.

The liquid separating device 5 comprises a liquid separating disc, 96 conical pipes 53 and a first porous plate 52; as shown in fig. 3, the first porous plate 52 is positioned in the liquid separating disc to divide the liquid separating disc into an upper part and a lower part, the upper part of the liquid separating disc is provided with a solvent bin 51 with a volume smaller than 2000 mu L, and the bottom of the first porous plate 52 is fixedly connected with the tops of 96 conical pipes 53 at the same time, so that the liquid in the solvent bin 51 can uniformly flow into each conical pipe 53; the conical tube 53 can be repeatedly cleaned and used, the upper opening of the conical tube is in a funnel-shaped design with the caliber of 400-500 mu m, the lower opening of the conical tube is matched with the caliber of the micro filling column 62, and preferably, the bottom of the conical tube 53 is also sleeved with a sealing ring 54, so that the conical tube 53 can be in sealing connection with the micro filling column 62, and the top of the liquid separation disc is connected with the bottom of the connector 4;

the filter device 6 comprises a filter disc 61, a porous plate II 63 and 96 micro-filling columns 62; as shown in fig. 4, the second perforated plate 63 is located in the filter disc 61 to divide the filter disc 61 into an upper part and a lower part, and 96 micro filling columns 62 are respectively installed on the perforations of the second perforated plate 63 in a penetrating manner; the tops of the 96 micro filling columns 62 are in one-to-one correspondence with the bottoms of the 96 conical tubes 53 and are detachably connected; in this embodiment, the micro-packed column 62 is made of pressure-resistant glass or quartz glass, has an inner diameter of 250 μm, an outer diameter of 360 μm, a length of 20cm, and a tip outlet of 20 μm, and is connected to the conical tube 53 via the seal ring 54 without creating a dead volume.

The micro filling column 62 is filled with a purifying filler 64, the filler length is 1/3 of the total length of the micro filling column 62, the sequence of filling the purifying filler 64 in the micro filling column 62 is that the filler with the particle diameter of 30 mu m is used for filling 1cm, after pressurized filling, the pressure can resist 500psi, and further the filler with the particle diameter of 5 mu m is used for filling 5cm, because the large filler forms an arch bridging function, the front filler forms a sieve plate-like function, the subsequent filler does not wash out the micro filling column 62 due to liquid phase, the corresponding micro filling column 62 can be inserted into a vacancy on the filter disc 61, the vacancy has a certain length, and the purifying filler 64 is one of silica gel, C18, diatomite and polystyrene-divinylbenzene; in this example, taking a C18 packing as an example, the whole device is connected, water is used as a solvent, a flow rate of 800-1200 microliters/min is realized, back pressure test is performed for fifteen times, average is 230psi, and long-term state can be maintained for more than 12 hours.

The liquid collecting device 7 comprises a base 71 and a plurality of liquid collecting bottles 72, and 96 liquid collecting bottles 72 are arranged on the base 71; the openings of the 96 liquid collection bottles 72 correspond to the outlet ends of the micro-fill column 62, respectively.

Example 2: a high-flux trace biological sample extracting solution purifying method, the embodiment uses 300 mu L serum as sample to carry out purifying operation;

s1: connecting a pump, a transfusion pipeline, a connector and a liquid separating device, starting a large flow rate of 2 mL/min, removing bubbles, and closing the pump when the outflow solution of the conical pipe is continuous fluid;

s2: the filter unit was connected to the liquid-separating unit, and then the pump was started to flow the purified filler C at a rate of 0.5 mL/min ₁₈ (coarse 30 μm, fine 5 μm particle size) of the packed column was subjected to methanol activation treatment, and then the pump was turned off;

s3: 300 mu L of serum sample is put in the activated micro-filling column, the filtering device is connected with the liquid separating device again, the pump is started for eluting, methanol water (60:40V/V) is used for eluting, the flow rate is 0.2 mL/min, the eluting time is 5 min, and meanwhile the eluent is collected in the liquid collecting bottle. The recovery rate of vitamin A in the sample detection is 97% -103% by contrast with the added sample.

Example 2: a high-flux trace biological sample extracting solution purifying method, in the embodiment, 150 mu L saliva is taken as a sample for purifying operation;

s1: connecting a pump, a transfusion pipeline, a connector and a liquid separating device, starting a large flow rate of 2 mL/min, removing bubbles, and closing the pump when the outflow solution of the conical pipe is continuous fluid;

s2: the filter unit was connected to the liquid-separating unit, and then the pump was started to flow the purified filler C at a rate of 0.5 mL/min ₁₈ (coarse 30 μm, fine 5 μm particle size) of the packed column was subjected to methanol activation treatment, and then the pump was turned off;

s3: and (3) placing 150 mu L of saliva sample in the activated micro-filling column, connecting the filtering device with the liquid separating device again, starting a pump to perform elution, eluting with methanol water (50:50V/V) at the flow rate of 0.2 mL/min for 3 min, and collecting the eluent in a liquid collecting bottle. The recovery rate of vitamin A in the sample detection is 93% -95% by contrast with the added sample.

The present embodiment is only for explanation of the present utility model and is not to be construed as limiting the present utility model, and modifications to the present embodiment, which may not creatively contribute to the present utility model as required by those skilled in the art after reading the present specification, are all protected by patent laws within the scope of claims of the present utility model.

Claims (4)

1. A high-flux trace biological sample extracting solution purifying device is characterized in that: the liquid storage device comprises a pump (3), a plurality of liquid delivery pipelines (8) and a plurality of liquid storage bottles (2), wherein filter elements (1) are arranged in the liquid storage bottles (2), the end parts of the liquid delivery pipelines (8) are connected with the filter elements (1), and the end parts of the liquid delivery pipelines (8) far away from the filter elements (1) penetrate through the liquid storage bottles (2) to be connected with the inlet ends of the pump (3); the outlet end of the pump (3) is provided with an infusion pipeline (9), the end, far away from the pump (3), of the infusion pipeline (9) is provided with a connector (4), the outlet end of the connector (4) is provided with a liquid separating device (5), the outlet end of the liquid separating device (5) is provided with a filtering device (6), and the outlet end of the filtering device (6) is provided with a liquid collecting device (7);

porous fibers (41) and a stainless steel screen plate (42) are arranged in the connector (4), and the bottom of the porous fibers (41) is fixedly connected with the top of the stainless steel screen plate (42);

the liquid separating device (5) comprises a liquid separating disc, a plurality of conical pipes (53) and a first porous plate (52); the first porous plate (52) is positioned in the liquid separation disc to divide the liquid separation disc into an upper part and a lower part, and the bottom of the first porous plate (52) is fixedly connected with the tops of the plurality of conical pipes (53) at the same time; the top of the liquid separation disc is connected with the bottom of the connector (4);

the filtering device (6) comprises a filtering disc (61), a porous plate II (63) and a plurality of micro filling columns (62); the second perforated plate (63) is positioned in the filter disc (61) to divide the filter disc (61) into an upper part and a lower part, and the plurality of micro filling columns (62) are respectively arranged on the perforations of the second perforated plate (63) in a penetrating way up and down; the tops of the micro filling columns (62) are in one-to-one correspondence with the bottoms of the conical tubes (53) and are detachably connected;

the micro-packed column (62) is filled with a purification packing (64).

2. The high throughput micro biological sample extraction liquid purification apparatus of claim 1, wherein: the liquid collecting device (7) comprises a base (71) and a plurality of liquid collecting bottles (72), and the liquid collecting bottles (72) are all arranged on the base (71); the openings of the liquid collection bottles (72) respectively correspond to the outlet ends of the micro filling columns (62).

3. The high throughput micro biological sample extraction liquid purification apparatus of claim 1, wherein: the purifying filler (64) is one of silica gel, C18, diatomite and polystyrene-divinylbenzene.

4. The high throughput micro biological sample extraction liquid purification apparatus of claim 1, wherein: sealing rings (54) are sleeved at the bottoms of the conical pipes (53).

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