JPS6150607A - Separation apparatus by chromatography - Google Patents

Abstract

PURPOSE:To automatically perform separating opertion so as to always obtain a predetermined recovery ratio, by performing the termination of recovery when the recovery ratio of the objective component flowed out from a column reaches the objective value or more. CONSTITUTION:An objective recovery ratio is inputted to an operation control part by an input device 12. Subsequently, a valve V1 is closed and a valve V2 is opened to flow solution to be separated to a separation agent packed column 6 while the valve V1 is opened and the valve V2 is closed to flow an eluate to the column 6. A valve V3 is closed and a valve V4 is opened and the recovery amount of the objective component flowed out from the column 6 is operated on the basis of the product of the outputs from a flow meter 4 and a concn. detector 7 and the elapse time and outlet concn. from the start point of the flow of the eluate by the operation control part 11. This calculated recovery ratio is compared with the recovery objective amount calculated from a preset recovery ratio and, at the point when the calculated recovery amount reaches the objective value or more, the valve V3 is opened and the valve V4 is closed to finish recovery.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a separation device by chromatography (hereinafter referred to as chromatography) using a column containing a separation packing material. A chromatography device configured to determine the boundary between the recovered fraction and the non-recovered fraction so that the recovery rate is determined, and to control means for separating the recovered fraction and the non-recovered fraction based on the determination result. The present invention relates to a separation device according to the present invention.

[Prior Art] Chromatography is used for the purpose of separating and purifying soluble substances, and is applied to separation and purification on an industrial scale in fields such as pharmaceuticals, foods, and fine chemistry.

In this chromatographic separation device, the recovered fraction (the recovered portion containing the target component in the liquid flowing out from the column) and the non-recovered fraction (the portion containing little or substantially no target component) are separated. It is important to accurately identify and separate.

In a conventional chromatographic separation device, the method for detecting the boundary between the recovered fraction and the non-recovered fraction of the liquid discharged from the separation column is as follows: Run it through the column under the same conditions as and find out its retention time.
Judge around this retention time using a timer. ■ Under conditions where the target component is completely separated from other components, judge based on the retention time of the target component. ■ Samples that are good even if the degree of separation is low. For this purpose, methods such as timer management are used to maintain empirically recognized retention times.

[Problems to be solved by the invention] However, in all of the above conventional methods, only the elapsed time since the sample was introduced into the separation column is determined, and the pH
1. Even if separation conditions such as salt concentration, temperature, flow rate, etc. change during separation operation, operation can only be performed under the conditions set before the change.

Unable to respond to change. Therefore, it was difficult to keep the recovery rate of the recovered fraction constant.

c. Means for Solving Problems] In order to solve the above problems, the present invention provides a solution to be separated and an eluent that are supplied to a column containing a separation packing material, and are passed through the column to collect the collected fraction. In an apparatus designed to separate fractions and unrecovered fractions, a target recovery rate of the target component is set in advance. Then, the arithmetic controller calculates the target amount of the target component to be recovered from the total amount of the target component introduced into the column and this target recovery rate. Further, the amount of liquid introduced into the column and the target component concentration in the liquid discharged from the column are input to the arithmetic controller. The arithmetic controller calculates the flow rate of the target component from the column based on the input data, calculates (integrates) the amount recovered up to that point, and compares the calculated recovery amount with the target recovery amount. When the actual recovery amount exceeds the target recovery difference, the means for separating the recovered fraction and the non-recovered fraction installed subsequent to the column is switched.

That is, the present invention comprises a column containing a separation packing material, a means for supplying a liquid to be separated and an eluent containing target components to the column, and a means for fractionating the liquid discharged from the column. , a concentration detector for detecting the concentration of a target component in a liquid, which is provided at least at an inlet and an outlet of the column, a flow rate detection means for detecting an inflow amount of the liquid to be separated into the column, and the concentration detection means. The output of the device and the flow rate detection means are input, and based on this, the flow rate of the target component from the column is calculated, and the recovery target of the target component is calculated from the total flow rate of the target component into the column and the preset recovery rate. The gist of the invention is a chromatographic separation device comprising: a calculation controller that calculates the amount of the target component flowing out and switches and controls the separation means when the amount of the target component flowing out exceeds the target amount of collection.

In the present invention, a flow meter may be used as the flow rate detection means, but if a metering pump is used as the liquid supply device to the column, the metering pump discharge amount is used as flow rate data, and this is sent to the arithmetic controller. You may also be asked to input it.

Further, in the present invention, the concentration detector may be provided only on the outlet side of the column, or may be provided on both the inlet side and the outlet side.

If the concentration detector on the inlet side is omitted, the concentration of the target component in the liquid to be separated should be analyzed automatically or manually in advance.
The results of this analysis are input into the arithmetic controller.

In the present invention, as the separation filler, any of various conventionally known fillers such as silica gel, alumina, ion exchange resins, active materials, etc. can be employed.

[Operation] According to the present invention, the recovery rate of the target component flowing out from the column ((amount of target component flowing out up to that point)/(
The separation operation is performed while detecting the total amount of the target component introduced into the column, and the flow path of the separation means is automatically switched to achieve a preset recovery rate.

[Example] Examples will be described below with reference to the drawings.

FIG. 1 is a system diagram of a chromatographic separation apparatus according to an embodiment of the present invention.

Reference numeral 1 denotes an eluent storage tank, which is connected to a separation agent-filled column 6 through a pipe 21 in which a liquid feed pump 3, a flow meter 4, and a concentration detector 5 on the inlet side are installed in this order.

Reference numeral 2 denotes a liquid storage tank to be separated, which is connected to a portion between the eluent storage tank l and the pump 3 via a pipe 22. V
1 and V2 are valves provided on the liquid outflow sides of the eluent storage tank 1 and the liquid to be separated storage tank 2, respectively.

A piping 23 for connecting to the one-separation means 8 is provided on the exit side of the separating agent-filled column 6, and a concentration detector 7 on the exit side is provided in the middle of this piping 23.

To explain the configuration of the fractionation means 8, the downstream part of the pipe 23 branches into pipes 24 and 25, the pipe 24 is provided with an outlet valve v3 for the non-recovered fraction, and the pipe 25 is provided with an outlet for the collected fraction. A valve v4 is provided, and a recovered fraction recovery tank 9 is provided downstream of the valve v4.

10 is an arithmetic controller that controls this device,
It has an arithmetic control section 11 and an input section 12 for inputting a preset recovery rate into the arithmetic control section 11.

Data signals are input to the calculation control section 11 from the flow meter 4, the concentration detector 5.7, and the input section 12, respectively. The arithmetic control unit 11 includes an A/D converter that converts these signals into digital signals, an input/output device, a central processing unit, a storage device that stores programs and data, a data bus that connects these, and the like. Further, the arithmetic control section 11 includes a D/A converter for outputting the arithmetic result as a control signal.

Note that the calculation control section 11 also has a built-in timer.

Next, the operation of the apparatus of the above embodiment will be explained, taking as an example a case where the development speed of both recovered fractions is higher than the development speed of the non-recovered fraction.

(2) First, the input device 12 inputs the target recovery rate to the arithmetic control section 11 .

■ Then, open valve vI, close valve v2, operate pump 3, and allow some eluent to flow into column 6 filled with separation agent, then close valve v1, open valve v2, and flow the liquid to be separated into column 6. , At this time, valve V3 is open and valve V4 is closed.

Detection signals are sent from the flow meter 4 and the detector 5 to the calculation control unit 11, and a timer built in the calculation control unit 11 measures the elapsed time from the start of the flow of the liquid to be separated.

■ After flowing the liquid to be separated through the column for a predetermined period of time, close the valve V.
+ is opened, v2 is closed, and the eluent is allowed to flow through column 6.

■ Next, close valve v3 and open ■4.

(2) The calculation control unit 11 calculates the total amount of the target component supplied to the column 6 based on the product of the outputs from the flowmeter 4 and the detector 5 and the time during which the liquid to be separated is allowed to flow.

■ The detection signal output value is also input from the outlet side detector 7 to the calculation control unit 11, and the calculation control unit 11 receives the output of the flowmeter 4, the elapsed time from the start of eluent flow, and the concentration at the exit side. By the product of
Calculate the cumulative amount (recovery amount) of the target component that has flowed out.

■ This calculated collection amount is compared with the collection target amount determined from the preset collection rate, and when the calculated collection amount is equal to or larger than the set collection target amount, the valve v3 is opened and 4 is closed to complete the collection of the collected fraction into the collection tank 9.

Note that since the above calculation example is an example of the present invention, the present invention is not limited thereto.

For example, after (2), the recovery amount may be divided by the total inflow amount of the target component to convert it into a recovery rate, and this recovery rate may be compared with the target recovery rate.

[Experimental Example 1] In the apparatus shown in Fig. 1, 1010
0O Yonokuichiru (il mark) HW manufactured by Toyo Soda Kogyo Co., Ltd.
-40c to a Pyrex glass column (inner diameter 50m)
m, length 500 mm), and fill the column 6 with packing material.
And so. As pump 3, an acid personal pump (metering pump) manufactured by Nippon Seimitsu Co., Ltd. was used. A differential refractometer was used as the detector 5.7.

As the liquid to be separated, a starch syrup solution adjusted to have a sugar concentration of 10% was used, and sugars containing no glucose were recovered.

Distilled water was used as the eluent.

Note that since a metering pump was used as the pump 3, this metering pump was used instead of the flow meter 4. The target recovery rate was set at 60%.

First, a target recovery rate of 0.6 was input from the input unit 12. After slightly flowing the eluent into column 6, close valve V+, open V2, close ■4, and open v3, and pass 100 mu of the above starch syrup solution at a flow rate of 4 or 1 minute, then close valve VI. With valve v4 open and valve v3 closed, the eluent (distilled water) was passed through at a rate of 40 m/min.

In the arithmetic control unit 11, the detected concentration signal from the detector 5.7 is read every 0.5 seconds, and the sugar concentration without glucose detected by the detector 5 is multiplied by the flow rate and the liquid flow time. to calculate the total wave person-borrow. And this is the target collection XJ
Compare the value multiplied by 0.6 and the integrated value (recovered amount) of the value detected from output 1 side detector 7 multiplied by 40 m of concentrated acid + 7 minutes, and compare the former value with the latter. When the values of V3. are equal to or greater than V3. I performed an operation to switch V4. The results are shown in Table 1.

In addition, tests were conducted in the same manner as in section ①- using starch syrup solutions in which the sugar concentration of the liquid to be separated was changed to 7% and 15%. The results are shown in Table 1.

Table 1 From Table 1, it is clear that sufficient separation was achieved with the recovery rate meeting the target value.

[Experimental Example 2] Using the same apparatus and the same packing material as those used in Experimental Example 1, separation operations were performed under the same conditions for sugar concentrations of 7% and 15%.

However, the recovery end point is determined by the detector 7 at the outlet of the separation force ram 6 when the recovery rate is 60% and the sugar concentration is 10% in Experimental Example 1.
When the output value of the detector 7 becomes below this value, the valve is switched between recovery and non-recovery. The results are shown in Table 2. In the method in which the recovery/non-recovery valve is switched when the value of the detector 7 on the outlet side of the separation column 6 becomes smaller than the set value, the recovery rate and the glucose elimination rate of the recovered liquid are as shown in Table 2. It fluctuated greatly.

Table 2 [Experimental Example 3] Using the same apparatus and packing material as those used in Experimental Example 1, a separation operation was performed under the same conditions at a sugar concentration of 7.15%.

However, the recovery end point is based on the time when the valve is switched between recovery and non-recovery when the recovery rate is 60% and the sugar concentration is 10% in Experimental Example 1. The switch was made. The results are shown in Table 3.

As shown in Table 3, when the recovery and non-recovery valve was operated using only the timer, it was observed that the recovery rate and the glucose elimination rate of the recovery liquid varied greatly as the concentration of the liquid to be separated changed.

Table 3 [Effects] As described above, the device of the present invention is configured to calculate the recovery rate of the target component flowing out from the column, and to terminate the recovery when this recovery rate exceeds the target recovery rate. The separation operation is automatically performed so that a predetermined recovery rate is always achieved.

[Brief explanation of the drawing]

FIG. 1 is a system diagram of a chromatographic separation apparatus according to an embodiment of the present invention. 1... Eluent storage tank, 2... Liquid to be separated storage tank, 3... Liquid sending pump, 4...
・Flowmeter, 5.7...Concentration detector, 6...Separating agent packed column, 8...Preparation device, 9...Recovered fraction Recovery tank, 10... Arithmetic controller.

Claims (1)

[Claims] (1) A column containing a separation packing material, a means for supplying a liquid to be separated containing a component to be separated and an eluent to the column, a means for fractionating the liquid discharged from the column, and the column. a concentration detector for detecting the concentration of a target component in the liquid, which is provided at least at an inlet and an outlet of the column; a flow rate detection means for detecting the amount of inflow of the liquid to be separated into the column; and the concentration detector and the flow rate. The output of the detection means is input, and based on this, the amount of the target component flowing out from the column is calculated, and the target amount of the target component to be recovered is calculated from the total amount of the target component flowing into the column and the preset recovery rate. and an arithmetic controller that switches and controls the separation means when the outflow amount of the target component exceeds the target recovery amount.