CN219111061U - A high-throughput micro-biological sample extraction liquid purification device - Google Patents

Abstract

The utility model discloses a purification device for extracting liquid of a high-throughput trace biological sample, which relates to the technical field of clinical examination and life science. Each of the liquid storage bottles is provided with a filter element, the ends of a plurality of infusion lines are connected to the filter element, and the ends of the plurality of infusion lines away from the filter element pass through the liquid storage bottle and are connected to the inlet port of the pump; The outlet end is provided with an infusion pipeline, the end of the infusion pipeline away from the pump is provided with a connector, the outlet end of the connector is provided with a liquid separator, and the outlet end of the liquid separator is provided with a filter device, and the filter The outlet end of the device is provided with a liquid collecting device. The device and method utilize the micro-level extraction and purification of the extract liquid under relatively high back pressure.

Description

A high-throughput micro-biological sample extraction liquid purification device

technical field

The utility model relates to the technical field of clinical examination and life science, more specifically, it relates to a high-throughput micro biological sample extraction liquid purification device.

Background technique

According to the needs of research, the detection of biological samples is mainly aimed at the determination of small molecule index substances, active ingredients, active ingredients, macromolecular composition, etc., to show the characteristics of the biological environment and biological process they represent, and to be used for physiological and biochemical research. . After the conventional sample is obtained, it generally goes through the steps of homogenization, extraction, purification, and detection. The commonly used purification methods include solid-phase extraction, liquid-liquid extraction, solid-liquid extraction, physical phase separation, etc., but in clinical and a large number of life science research Among them, due to the limitations and trace requirements of samples, the common extraction and purification methods require a large amount of samples, and it is difficult to meet the purification requirements of trace samples. In the case of low concentrations of target substances, This purification contradiction is particularly prominent, resulting in inaccurate test values and large test errors. In response to these problems, microextraction methods have been developed in recent years, such as solid phase microextraction and solid dispersion extraction.

Among them, the solid phase microextraction (SPME) technology is based on the use of fused silica fibers coated with a stationary phase to absorb and enrich the analyte in the sample. The biggest feature is that it can concentrate the analyte while extracting, but it can be seen The mainstream adsorption extraction method has the problem of re-sampling part of the sample. In addition, the selectivity of the filler will also affect the enrichment and accuracy of the effective substance in the sample, which will lead to inaccurate test results and poor quantitative results. Difference.

At the same time, the traditional SPME technology currently on the market can only process one sample at a time, and the throughput used is limited; in addition, the purification of the solid dispersion method commonly used in recent years also has certain requirements for the sample volume, and there is a solution for trace liquid samples. Due to the limitation of processing, it is difficult to effectively recover or enrich the corresponding target in a uniform environment, resulting in inaccurate detection.

Utility model content

The purpose of the utility model is to provide a high-throughput micro biological sample extraction liquid purification device, the device and method utilizes the micro-level extraction and purification of the extraction liquid under relatively high back pressure.

The above-mentioned technical purpose of the utility model is achieved through the following technical solutions: a high-throughput micro biological sample extraction liquid purification device, including a pump, a plurality of infusion pipelines and a plurality of liquid storage bottles, and a plurality of said liquid storage bottles There are filter elements inside, the ends of a plurality of infusion pipelines are connected to the filter elements, and the ends of the plurality of infusion pipelines away from the filter elements pass through the liquid storage bottle and are connected to the inlet end of the pump; the outlet end of the pump is provided with An infusion pipeline, the end of the infusion pipeline away from the pump is provided with a connector, the outlet end of the connector is provided with a liquid distributing device, the outlet end of the liquid dispensing device is provided with a filter device, and the outlet end of the filter device Equipped with a liquid collection device;

A porous fiber and a stainless steel sieve are arranged inside the connector, and the bottom of the porous fiber is fixedly connected to the top of the stainless steel sieve;

The liquid separator includes a liquid separator, a plurality of conical tubes and a porous plate; The top of a tapered tube is fixedly connected; the top of the liquid separator plate is connected with the bottom of the connector;

The filter device includes a filter disc, a porous plate 2 and a plurality of micro-packed columns; the porous plate 2 is located in the filter disc to separate the filter disc into upper and lower parts, and a plurality of the micro-filled columns are respectively installed on the porous plate up and down. On the second perforation; the tops of the plurality of micro-filled columns correspond to the bottoms of the plurality of tapered tubes and are detachably connected;

The micro packing column is filled with purification packing.

Further: the liquid collection device includes a base and a plurality of liquid collection bottles, and the plurality of liquid collection bottles are installed on the base; the openings of the plurality of liquid collection bottles correspond to the outlet ends of the micro-filling column respectively.

Further: the purification filler is one of silica gel, C ₁₈ , diatomaceous earth, and polystyrene-divinylbenzene.

Further: the bottoms of the plurality of tapered tubes are all sleeved with sealing rings.

A method for purifying a high-throughput trace biological sample extract, characterized in that it specifically includes the following steps:

S1: Connect the pump, infusion line, connector and dispensing device, turn on the high flow rate, remove air bubbles, and turn off the pump when the solution flowing out of the conical tube is continuous fluid;

S2: Connect the filter device to the liquid separation device, then turn on the pump, and turn off the pump after activating the pre-filled purification packing;

S3: Put the extract on the activated micro-packed column, connect the filter device to the liquid separation device again, turn on the pump for elution, and collect the eluate in the liquid collection bottle at the same time.

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

1. The purification device uses the elution and purification method of the sample pass-through under a relatively high pressure environment, which increases the selectivity of the target substance on the one hand, and reduces the loss of the sample target substance on the other hand;

2. Filling the purification packing in the micro-packing column can use the arch bridging effect formed by the packing with different particle sizes, and form a pressure-resistant filling environment under the one-way fluid, abandoning the use of the sieve plate, and in addition Under higher pressure, the compressibility and polarity of the fluid itself will change, and the material distribution relationship formed between it and the solid filler will change, which improves the extraction efficiency of the material;

3. The sample adopts the all-through elution method, which effectively solves the transfer loss of the sample, improves the recovery rate of the target substance, and the accuracy of the subsequent detection will also increase accordingly.

Description of drawings

Fig. 1 is a schematic structural diagram of a high-throughput trace biological sample extraction solution purification device in Example 1 of the present invention;

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

Fig. 3 is a schematic structural view of the liquid separator in Example 1 of the utility model;

Fig. 4 is the structural representation of filter device in the utility model embodiment 1;

Fig. 5 is the schematic structural view of the trace packing column in the utility model embodiment 1;

Fig. 6 is a schematic structural view of the liquid collection device in Embodiment 1 of the present utility model.

In the figure: 1. Filter element; 2. Liquid storage bottle; 3. Pump; 4. Connector; 41. Porous fiber; 42. Stainless steel sieve plate; 53. Conical tube; 54. Sealing ring; 6. Filtration device; 61. Filter plate; 62. Trace packing column; 63. Porous plate 2; 64. Purification filler; Liquid collection bottle; 8. Infusion pipeline; 9. Infusion pipeline.

Detailed ways

Below in conjunction with accompanying drawing 1-6 the utility model is described in further detail.

Embodiment 1: A high-throughput microbiological sample extraction liquid purification device, as shown in Figures 1 to 6, includes a pump 3, 2 infusion lines 8 and 2 liquid storage bottles 2, and 2 liquid storage bottles 2 Both are equipped with a filter element 1, and the filter element 1 is connected to the inlet end of the pump 3 through the infusion line 8; The infusion pipeline 9 is connected. In the present embodiment, the infusion pipeline 8 and the infusion pipeline 9 are preferably peek tubes or stainless steel pipes with a narrow inner diameter (165-360 μm), and the outlet end of the connector 4 is fixedly connected to the liquid dispensing device 5. The outlet end of the device 5 is detachably connected to the inlet end of the filter device 6, the liquid collecting device 7 is located at the outlet end of the filter device 6, and the liquid collecting port corresponds to the outlet of the filter device 6;

Porous fiber 41 and stainless steel sieve plate 42 are installed in the connector 4, as shown in Figure 2, the bottom of porous fiber 41 is fixedly connected with the top of stainless steel sieve plate 42; Maintain back pressure and achieve effective liquid dispensing, eliminating air bubbles.

Liquid separator 5 comprises liquid separator, 96 conical tubes 53 and perforated plate one 52; As shown in Figure 3, perforated plate one 52 is positioned at liquid separator and divides liquid separator into upper and lower two parts, and liquid separator top It is a solvent chamber 51 with a volume of less than 2000 μL. The bottom of the porous plate 1 52 is fixedly connected to the tops of 96 conical tubes 53 at the same time, so that the liquid in the solvent chamber 51 can evenly flow into each conical tube 53; the conical tubes 53 Can be used repeatedly for cleaning, the upper port is a funnel-shaped design with a caliber of 400-500 μm, and the lower port is adapted to the caliber of the micro-packed column 62. Preferably, a sealing ring 54 is also sleeved at the bottom of the conical tube 53, so that the The tapered tube 53 is tightly connected to the micro-filling column 62, and the top of the liquid separator is connected to the bottom of the connector 4;

Filtration device 6 comprises filter disc 61, porous plate 2 63 and 96 micro-packed columns 62; As shown in Figure 4, porous plate 2 63 is positioned at filter disc 61 and filter disc 61 is divided into upper and lower two parts, 96 micro-filled columns 62 are respectively installed up and down on the perforations of the porous plate 2 63; the tops of the 96 micro-filled columns 62 correspond to the bottoms of the 96 conical tubes 53 and are detachably connected; in this embodiment, the micro-filled columns 62 are Made of pressure-resistant glass or quartz glass, with an inner diameter of 250 μm, an outer diameter of 360 μm, a length of 20 cm, and a tip outlet of 20 μm, it is connected to the tapered tube 53 through a sealing ring 54 without dead volume.

The micro-packed column 62 is filled with purification filler 64, and the length of the filler accounts for 1/3 of the total length of the micro-packed column 62. The order of filling the purification filler 64 in the micro-packed column 62 is to fill 1 cm with a filler of 30 μm particle size. , can withstand a pressure of 500psi, and then use fillers with a particle size of 5μm to fill 5cm. Since the large fillers form an arched bridging effect, the front-end fillers form a sieve-like effect, and the subsequent fillers will not be flushed due to the liquid phase. A micro-packed column 62 is produced, and the corresponding micro-packed column 62 can be inserted into the vacancy on the filter disc 61. The vacancy has a certain length, and the purification packing 64 is one of silica gel, C18, diatomaceous earth, polystyrene-divinylbenzene ; In this embodiment, take C18 packing as an example, connect the overall device, use water as the solvent, realize the flow rate of 800-1200 microliters/minute, back pressure test fifteen times, the average is 230psi, and the long-term state can be maintained within 12 hours above.

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

Embodiment 2: A high-throughput method for purifying microbiological sample extracts. In this embodiment, 300 μL of serum is used as the sample to perform the purification operation;

S1: Connect the pump, infusion line, connector and dispensing device, turn on the maximum flow rate of 2mL/min, remove air bubbles, and turn off the pump when the solution flowing out of the conical tube is a continuous fluid;

S2: Connect the filter device to the liquid separation device, and then turn on the pump so that the flow rate is 0.5mL/min. After the micro-packed column of the pre-filled purification filler C ₁₈ (30 μm thick, 5 μm particle size) is activated by methanol , turn off the pump;

S3: Put 300 μL serum sample on the activated micro-packed column, connect the filter device to the separator again, turn on the pump for elution, and elute with methanol water (60:40V/V) at a flow rate of 0.2mL /min, elute for 5 minutes, and collect the eluate in the collection bottle at the same time. The vitamin A in the sample detection is compared with the added sample, and the recovery rate is 97%-103%.

Embodiment 2: A high-throughput purification method for microbiological sample extracts. In this embodiment, 150 μL of saliva is used as the sample for purification;

S1: Connect the pump, infusion line, connector and dispensing device, turn on the maximum flow rate of 2mL/min, remove air bubbles, and turn off the pump when the solution flowing out of the conical tube is a continuous fluid;

S2: Connect the filter device to the liquid separation device, and then turn on the pump so that the flow rate is 0.5mL/min. After the micro-packed column of the pre-filled purification filler C ₁₈ (30 μm thick, 5 μm particle size) is activated by methanol , turn off the pump;

S3: Put 150 μL of saliva sample on the activated micro-packed column, connect the filter device to the liquid separation device again, turn on the pump for elution, and elute with methanol water (50:50V/V) at a flow rate of 0.2mL / minute, eluted for 3 minutes, and the eluate was collected in the collection bottle at the same time. The vitamin A in the sample detection is compared with the added sample, and the recovery rate is 93%-95%.

This specific embodiment is only an explanation of the utility model, and it is not a limitation of the utility model. After reading this description, those skilled in the art can make modifications to this embodiment without creative contribution according to needs, but as long as it is within the utility model All new claims are protected by patent law.

Claims (4)

1. A high-throughput trace biological sample extraction liquid purification device is characterized in that: comprising a pump (3), a plurality of infusion lines (8) and a plurality of liquid storage bottles (2), a plurality of said liquid storage bottles ( 2) There are filter elements (1) inside, the ends of a plurality of infusion pipelines (8) are connected to the filter elements (1), and the ends of the plurality of infusion pipelines (8) away from the filter elements (1) pass through the storage The liquid bottle (2) is connected to the inlet end of the pump (3); the outlet end of the pump (3) is provided with an infusion pipeline (9), and the end of the infusion pipeline (9) away from the pump (3) is provided with a connection device (4), the outlet end of the connector (4) is provided with a liquid separator (5), the outlet end of the liquid separator (5) is provided with a filter device (6), and the filter device (6) The outlet end is provided with a liquid collecting device (7); The connector (4) is provided with a porous fiber (41) and a stainless steel sieve plate (42), and the bottom of the porous fiber (41) is fixedly connected with the top of the stainless steel sieve plate (42); The liquid separator (5) comprises a liquid separator, a plurality of conical tubes (53) and a porous plate one (52); the porous plate one (52) is located in the liquid separator and separates the liquid separator into upper and lower two part, the bottom of the porous plate one (52) is fixedly connected with the tops of a plurality of conical pipes (53) at the same time; the top of the liquid separator is connected with the bottom of the connector (4); Described filtering device (6) comprises filter disc (61), porous plate two (63) and a plurality of micro-packed columns (62); Described porous plate two (63) is positioned at filter disc (61) and filters disc (61) ) is divided into upper and lower parts, and a plurality of micro-packed columns (62) are installed up and down on the perforated plate two (63) respectively; The bottoms of (53) correspond one-to-one and are detachably connected; The micro packing column (62) is filled with purification packing (64). 2. A high-throughput purification device for trace biological sample extraction liquid according to claim 1, characterized in that: the liquid collection device (7) includes a base (71) and a plurality of liquid collection bottles (72), more Each of the liquid collection bottles (72) is installed on the base (71); the openings of the plurality of liquid collection bottles (72) correspond to the outlet ends of the micro-packed column (62) respectively. 3. A kind of high-throughput micro biological sample extraction liquid purification device according to claim 1, characterized in that: said purification filler (64) is made of silica gel, C18, diatomaceous earth, polystyrene-divinylbenzene one of them. 4. A high-throughput purification device for trace biological sample extraction liquid according to claim 1, characterized in that: the bottoms of the plurality of tapered tubes (53) are all sleeved with sealing rings (54).

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