JP4968815B2 - Emulsion automatic separation method - Google Patents

Description

  The present invention relates to a method for separating an emulsion into two phases through a filter.

  In research and development in the fields of food, medicine, cosmetics, paints, adhesives, oils, etc., liquids that do not dissolve in each other may be stirred and mixed to be emulsified, and then separated into two phases again. For example, it is a process necessary for liquid-liquid extraction for crude purification of a synthetic product, extraction of a drug in blood, or extraction of residual agricultural chemicals from agricultural products. In that case, instead of the centrifugal separation method, Patent Document 1 and Patent Document 2 disclose a method of separating the emulsion into two phases through a filter.

  In patent document 1, the emulsion which consists of water and a polycarbonate solution is isolate | separated into the aqueous solution phase and the polycarbonate solution phase by passing through the filter whose contact angle with water is 40 degrees or less.

  In Patent Document 2, an emulsion containing oil in a liquid is separated through a first separator and a second separator each composed of a plurality of layers of filters each having a fiber diameter and fiber density changed stepwise. Specifically, the liquid to be treated is passed through the first separator from the larger fiber diameter and the smaller fiber density toward the smaller fiber diameter and the smaller fiber density to remove coarse dust and oil to some extent. To enlarge. Next, the emulsion is separated into two phases by passing the liquid to be treated through the second separator from the smaller fiber diameter and the smaller fiber density toward the larger fiber diameter and the coarse fiber density.

Japanese Examined Patent Publication No. 46-41622 (third column) JP-A-8-309102 (paragraph numbers 0020 to 0021, FIG. 2)

  The inventions described in Patent Documents 1 and 2 are suitable for uniformly processing a specific emulsion in large quantities, but may not be suitable for processing a small amount of various test solutions. In laboratories, etc., a wide variety of emulsions are handled for the development of new drugs and detection of contained components, and the amount handled is small. In each test, it is necessary to replace the container and filter for each test in order to prevent mixing of different test solutions. However, in Patent Documents 1 and 2, once the device is assembled, the filter is replaced unless clogging or aging occurs. This is because it cannot be applied to the separation method at the laboratory level.

  Most of the test operations in laboratories are performed manually, and the test procedure for each reagent is basically the same. Therefore, the researcher needs a lot of time and labor just by repeating simple operations such as dispensing of the test solution, which is very complicated. Although it was a simple task, a certain level of skill was required to ensure the accuracy of the data, and there was a waste that a competent researcher was restrained by a complicated task.

  Accordingly, an object of the present invention is to automate and unmanned a series of operations such as dispensing of a wide variety of small amounts of test solutions and separation of emulsions, and to reduce the cost required for testing. An object of the present invention is to obtain a highly accurate data by mechanizing and always creating a constant reagent.

In the method for automatically separating an emulsion of the present invention, an emulsified reagent solution is passed through a filter to separate into two phases, and an arm 4 that can move in the XYZ directions on the processing table 2 and a reagent solution dispensing provided on the arm 4 a probe 5 of use, has an automatic dispensing device 1 comprising a dispensing means for discharging the test solution from the probe 5 suction-holding the reagent in probe 5.

As shown in FIG. 3, the separation method includes the first step of dispensing the dispersion medium 16 from the dispersion medium container 15 to the emulsion container 7 by the probe 5, and the dispersion 18 from the dispersion container 17 by the probe 5. The second step of dispensing into the container 7, the third step of stirring and mixing the dispersion medium 16 and the dispersion liquid 18 separated into two upper and lower phases in the emulsion container 7, and emulsifying the emulsion by the probe 5. and the reduction have been reagent from the emulsion vessel 7 to the separation vessel 11, by dispenses through a filter 13 disposed on the separation container 11 min, a dispersion medium 16 and emulsified the reagent in the separation vessel 11 dispersed And a fourth step of re-separating the liquid 18 into two phases . In the second process diagram of FIG. 3, the dispersion medium 16 and the dispersion liquid 18 are stacked in the emulsion container 7 downward, and depending on the specific gravity of the dispersion medium 16 and the dispersion liquid 18, this may be reversed upside down. Sometimes.

  In each of the first to fourth steps, the probe 5 is washed by the rinse station 20 before moving to the next step. Emulsion is composed of two liquid phases that do not dissolve each other, and one liquid phase (dispersion) is dispersed in the form of fine particles in the other liquid phase (dispersion medium), and is also called an emulsion.

  According to the present invention, since a series of operations from the first to the fourth steps ranging from the dispensing of the reagent solution to the separation of the emulsion can be automatically performed, it is not necessary for researchers to be involved in this operation. Human costs can be greatly reduced. Since the dispensing amount and the like can be accurately measured by the dispensing means of the automatic dispensing device, a constant amount of the test solution can be always dispensed, and highly accurate data can be obtained.

  If the residual reagent adhering to the inner and outer surfaces of the probe 5 is washed away by the rinsing station 20 before proceeding to the next process, the other test liquids are not mixed with the reagent used in each process, which is advantageous for obtaining accurate data. It is.

  1 and 2 show an automatic dispensing apparatus 1 used for carrying out the method of the present invention, which is arranged adjacent to a processing table 2. The automatic dispensing apparatus 1 includes a main body 25 that drives the arm 4, an arm 4 that is connected to the main body 25 and moves on the processing table 2 in the XYZ directions, and a hollow needle-like stainless steel probe provided on the arm 4. 5 and a dispensing means for applying a positive pressure or a negative pressure to the probe 5 to suck and hold the test solution on the probe 5 or to discharge the test solution from the probe 5. The dispensing means uses a three-way valve syringe pump (not shown), and the three-way valve syringe pump and the probe 5 are connected by a flexible tube.

  The operation of these devices is controlled by a computer, and dedicated program software for controlling the operation procedure of the automatic dispensing apparatus 1 over time is incorporated. Before conducting the test, the dispensing procedure is determined by inputting the test parameters such as the type of reagent solution, its containing position, and the dispensing volume with a keyboard, for example, and the arm 4 can be placed at any place in any dispensing process. By moving the sample, the probe 5 can be accurately sucked and held by the probe 5, and then the held reagent can be discharged.

  On the processing table 2, an emulsion production rack 8 that holds the emulsion container 7, a stirrer 9 that is installed below the emulsion production rack 8 and that stirs and mixes the test solution in the emulsion container 7 to produce an emulsion, and a separation container 11, a filter rack 12 that holds the dispersion medium 16, a dispersion medium container 15 that contains the dispersion medium 16, a dispersion liquid container 17 that contains the dispersion liquid 18, and a rinse station 20 that cleans the probe 5 are arranged at arbitrary positions. It is. The arrangement of the devices in FIGS. 1 and 2 is an example, and other different arrangements may be used. This is because all the movements of the arm 4 are controlled by the computer as described above, so that the movement of the arm 4 can be set in accordance with the arrangement.

  A plurality of emulsion containers 7 and separation containers 11 are held upright in the emulsion production rack 8 and the filter rack 12, and a stirrer is placed in each emulsion container 7. The stirrer 9 incorporates a plurality of magnets corresponding to the positions of the respective emulsion containers 7. When the magnets are rotated, the stirrer in the emulsion container 7 rotates, and the sample solution in the emulsion container 7 is stirred and mixed. it can.

  A filter is disposed above each separation container 11 on the filter rack 12. Specifically, in the fourth process diagram of FIG. 3, a circular cylindrical portion fitted into the upper end of the opening of the separation container 11, a flange provided at the outer peripheral edge of the upper end of the cylindrical portion, and a narrow tube continuously provided at the lower end of the cylindrical portion And a disc-like filter 13 is arranged in the cylindrical portion of the column 14. The column 14 is mounted inside the upper end of the separation container 11 from above, and an insertion hole through which the probe 5 is inserted in close contact is provided in the center of the upper lid.

  A rinsing station 20 is provided on the processing table 2, and a container containing cleaning water is disposed here. The inner and outer surfaces of the probe 5 are cleaned by an overflow method in which the probe 5 enters the rinse station 20 and sucks and discharges the cleaning water.

<Preparation process>
The emulsion production rack 8 and the filter rack 12 on the processing table 2 are set with the required number, that is, the type of the test solution and the number of test emulsion containers 7 and the separation containers 11, and the test solution is provided above each separation container 11. Install the filter. The dispersion medium container 15 and the dispersion liquid container 17 may be of a single type as shown in FIGS. 1 and 2 in the case where a certain amount of the test solution is mixed by changing the blending amount. Arrange everything. Next, the moving process and moving location of the arm 4 in each process, the dispensing amount of the reagent, and the like are input to the program software of the computer. Although these operations are performed manually, the automatic dispensing apparatus 1 performs the subsequent first to fourth steps.

<First step>
First, the arm 4 of the automatic dispensing apparatus 1 moves onto the dispersion medium container 15, and a predetermined amount of the dispersion medium 16 is sucked and held by the probe 5 by the dispensing means, and the dispersion medium 16 is moved from the probe 5 onto the emulsion generation rack 8. Discharge into one emulsion container 7. By repeating this, a predetermined amount of the dispersion medium 16 in the dispersion medium container 15 is sequentially dispensed into each emulsion container 7 of the emulsion production rack 8. When the dispersion medium 16 has been dispensed into all the emulsion containers 7, the probe 5 is washed by the rinse station 20.

<Second step>
Next, the arm 4 moves onto the dispersion liquid container 17, and a predetermined amount of the dispersion liquid 18 is sucked and held by the probe 5 by the dispensing means, and the dispersion liquid 18 is drawn from the probe 5 to one emulsion container on the emulsion production rack 8. 7 to discharge. By repeating this, the dispersion 18 in the dispersion container 17 is sequentially dispensed into each emulsion container 7 of the emulsion production rack 8 by a predetermined amount. The dispensing order in the second step is the same as the dispensing order in the first step. When the dispersion liquid 18 has been dispensed into all the emulsion containers 7, the probe 5 is washed again at the rinse station 20.

<Third step>
When dispensing of the dispersion medium 16 and the dispersion liquid 18 into each emulsion container 7 is completed, the stirrer 9 is activated and the dispersion medium 16 and the dispersion liquid 18 in the emulsion container 7 are stirred and mixed to be emulsified. The start timing and agitation time of the stirrer 9 are also controlled by computer program software, and activation, agitation, and stop are performed fully automatically.

<4th process>
When the dispersion medium 16 and the dispersion liquid 18 in the emulsion container 7 are emulsified by the stirrer 9, after the entire emulsion 19 in the emulsion container 7 is sucked and held by the probe 5, the probe 5 is inserted into the insertion hole in the upper lid of the column 14. Then, the emulsion 19 is discharged into the column 14, and the emulsion 19 is dispensed into the separation container 11 through the filter 13. When the probe 5 is inserted into the insertion hole of the upper lid of the column 14, the column 14 is sealed, and when the emulsion 19 is injected therein, the internal pressure of the column 14 rises and the emulsion 19 passes through the filter 13 in a pressurized state. Thus, the emulsion 19 is separated into the dispersion medium 16 and the dispersion liquid 18 in the separation container 11. Further, since the column 14 and the separation container 11 are not in close contact with each other, an escape route for the internal air of the separation container 11 is also secured.

(Test example)
Using the above-described automatic test apparatus, a test solution for measuring a drug component contained in horse serum and a test solution for a pigment amount test of orange juice were prepared. In this test example, the dispersion 16 is horse serum and orange juice of 100% fruit juice, and the dispersion medium 18 is n-hexane and chloroform for horse serum and 2,2,4-trimethyl for orange juice. Each pentane was used. The orange juice was adjusted to pH by adding 0.5 ml of hydrochloric acid to 5 ml of juice during the test.

<Preparation process>
Three vials were prepared as the emulsion container 7 and three test tubes were prepared as the separation containers 11, respectively, and set at predetermined positions of the emulsion production rack 8 and the filter rack 12. The filter 13 installed on the test tube 11 includes an incubation column (manufactured by Intavis) with a PTFE 20 μm pore filter for an n-hexane dispersion medium emulsion, and a PTFE 1 μm pore filter for a chloroform dispersion medium emulsion. A built-in column (Whatman) was used for the orange juice emulsion, and an incubation column (Intavis) with a 20 μm pore filter made of PE was used.

<First to fourth steps>
On the processing table 2, three test solution bottles containing n-hexane, chloroform and 2,2,4-trimethylpentane as the dispersion medium container 15 are arranged, and a test solution containing horse serum as the dispersion container 17. One bottle and one vial containing pH-adjusted orange juice were placed. After that, when the automatic dispensing apparatus 1 is started by inputting necessary data such as the moving procedure of the arm 4 and the dispensing amount of each reagent into the program software, the above first to fourth steps are automatically performed and the emulsion is formed. It was separated into two phases fully automatically. In this test example, in the first, second, and fourth steps, the probe 5 was washed with the rinse station 20 for each dispensing, in addition to before moving to the next step.

  After all the processes were completed, the filter 13 on the test tube 11 was removed by hand, the dispersion medium 18 was taken out with a dropper, and each test of the next step such as measurement of the contained drug amount was performed.

  In order to evaluate the reliability of the automatic separation method of the present invention, the inventors compared with the case where the emulsion was separated by a centrifuge. As a result, according to the present invention, it was confirmed that the emulsion was clearly separated into two phases as much as or more than when separated by a centrifuge.

  In the above-described embodiment, the arm 4 composed of three arms movable along the X axis, the Y axis, and the Z axis is illustrated, but this is not necessary. For example, a single arm used in an industrial robot or the like may be movable in the XYZ directions.

Perspective view of apparatus used in the present invention Schematic plan view showing an arrangement example of devices on the processing table Conceptual diagram showing each process

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Automatic dispensing apparatus 2 Processing table 4 Arm 5 Probe 7 Emulsion container 8 Emulsion production rack 9 Stirrer 11 Separation container 12 Filter rack 15 Dispersion medium container 17 Dispersion liquid container 20 Rinse station

Claims (2)

In order to pass the emulsified reagent through a filter and separate it into two phases, an arm (4) movable in the XYZ directions on the processing table (2), and a probe for dispensing the reagent provided on the arm (4) ( 5) comprises a probe (automatic dispensing device containing from 5) a reagent suction retained probe (5) and a dispensing means for discharging the reagent (1),
A first step of dispensing the dispersion medium (16) from the dispersion medium container (15) to the emulsion container (7) by the probe (5);
A second step of dispensing the dispersion (18) from the dispersion container (17) into the emulsion container (7) by means of the probe (5);
A third step of emulsifying the dispersion medium (16) and the dispersion liquid (18), which are separated into upper and lower two phases in the emulsion container (7), by stirring and mixing;
By the probe (5), and the emulsified been reagent from the emulsion vessel (7) to the separation vessel (11), by dispensing through a filter disposed on the isolation container (11) (13), an emulsion reduction has been reagent automatic separation method of an emulsion consisting of a fourth step of Ru re separate into two phases of the dispersion and the dispersion medium (16) in the separation vessel (11) (18).   The method for automatically separating emulsions according to claim 1, wherein in each of the first to fourth steps, the probe (5) is washed by a rinse station (20) before proceeding to the next step.

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