JP4431465B2 - Sample supply method and sample supply apparatus in supercritical fluid chromatography - Google Patents

Description

  The present invention relates to a method and apparatus for supplying a sample to a mobile phase in a supercritical fluid chromatography apparatus.

  In a supercritical fluid chromatography apparatus, a supercritical fluid is used as a mobile phase. A supercritical fluid has higher diffusibility and lower viscosity than a normal solvent. For this reason, in the supercritical fluid chromatography apparatus, for example, optical isomers that are difficult to separate can be separated at high speed.

  In a supercritical fluid chromatography device, an injection device such as an injector that injects a sample into a mobile phase is usually used. A supercritical fluid is a fluid that exists at an appropriate pressure and temperature. For this reason, in a supercritical fluid chromatography device, if pressure fluctuations occur during sample injection, the composition of the mobile phase changes, which may adversely affect the separation of the target substance.

  As a technique for preventing pressure fluctuation at the time of injection of a sample in a supercritical fluid chromatography apparatus, for example, a sample is supplied to a sample loop by a sample pump against a back pressure of a mobile phase containing a supercritical fluid, A technology that increases the pressure of the sample in the sample loop to the pressure of the mobile phase, then connects the sample loop to the flow path of the mobile phase, injects the sample in the sample loop into the flow path of the mobile phase, and introduces it to the column It is known (for example, refer to Patent Document 1).

The above technique is excellent from the viewpoint of preventing pressure fluctuations during sample injection, but it requires the use of pressure transmission means and the complexity of the channel shape in order to transmit the pressure of the mobile phase to the sample before injection. However, there remains room for study from the viewpoint of application to an existing supercritical fluid chromatography apparatus.
Japanese Patent Laid-Open No. 5-3007026

  It is an object of the present invention to supply a sample to a mobile phase with a simple configuration without pressure fluctuation in a supercritical fluid chromatography apparatus.

  In the present invention, when a mobile phase containing a supercritical fluid and a solvent is produced, a sample or a sample is replaced with a solvent which is pressed into a mobile phase flow path from a normal pressure state, or in place of the solvent. A method and apparatus for supplying a predetermined amount of a sample solution containing a solvent and supplying a sample to a mobile phase are provided.

  That is, the present invention relates to a sample supplied to a mobile phase containing a supercritical fluid and a solvent, and in supercritical fluid chromatography that separates a target substance from a sample containing the target substance. Supercritical fluid chromatography is a method of mixing a liquefied gas or a supercritical fluid and a solvent for constituting a supercritical fluid in a mobile phase flow path whose pressure is adjusted. In the method of supplying a sample, a predetermined amount of a sample solution containing a sample and a solvent is intermittently connected to the mobile phase flow path instead of the solvent mixed with the liquefied gas or the supercritical fluid in the mobile phase flow path. To provide a method of supplying automatically.

In supercritical fluid chromatography, the mobile phase is stabilized in the mobile phase flow path by mixing a lower pressure (usually normal pressure) solvent with a liquefied gas or supercritical fluid that is at a higher pressure than the solvent. Is generated. Therefore, in the above method, instead of the solvent mixed with the liquefied gas or the supercritical fluid, a predetermined amount of the sample solution is supplied to the liquefied gas or the supercritical fluid by the same method as the solvent, so that the sample is contained. The mobile phase is stably generated in the mobile phase channel. Therefore, the pressure fluctuation of the mobile phase accompanying the sample supply is prevented.

  The present invention also relates to a supercritical fluid chromatography apparatus for separating a target substance from a sample supplied to a mobile phase containing a supercritical fluid and a solvent and containing the target substance. The supercritical fluid chromatography apparatus is configured to supply a mobile phase channel, a pressure adjusting unit for adjusting the pressure of the mobile phase channel, and a pressure by the pressure adjusting unit. Mobile phase generation means for generating a mobile phase containing a supercritical fluid and a solvent in the adjusted mobile phase flow path, and the mobile phase generation means includes a liquefied gas for constituting the supercritical fluid or Supercritical fluid high pressure pump for supplying supercritical fluid to mobile phase flow path, solvent high pressure pump for supplying solvent to mobile phase flow path, and solvent high pressure pump for supplying solvent Suction side flow for solvent Has bets, apparatus for supplying samples, a sample solution of a predetermined amount containing a sample and a solvent to provide an apparatus having a means for supplying the suction side flow passage solvent.

  In the apparatus, since a predetermined amount of sample is supplied to the solvent mixed with the liquefied gas or supercritical fluid, the mobile phase containing the sample is stably generated in the mobile phase flow path as in the above method. Thus, fluctuations in the pressure of the mobile phase accompanying the sample supply are prevented. In addition, since the sample is appropriately supplied to the solvent flow path with respect to the mobile phase flow path, the sample is supplied with a simpler configuration.

  The present invention also relates to a supercritical fluid chromatography apparatus for separating a target substance from a sample supplied to a mobile phase containing a supercritical fluid and a solvent and containing the target substance. The supercritical fluid chromatography apparatus is configured to supply a mobile phase channel, a pressure adjusting unit for adjusting the pressure of the mobile phase channel, and a pressure by the pressure adjusting unit. Mobile phase generation means for generating a mobile phase containing a supercritical fluid and a solvent in the adjusted mobile phase flow path, and the mobile phase generation means includes a liquefied gas for constituting the supercritical fluid or The supercritical fluid high pressure pump for supplying the supercritical fluid to the mobile phase flow path, the solvent high pressure pump for supplying the solvent to the mobile phase flow path, and the solvent discharged from the solvent high pressure pump Supply to mobile phase channel An apparatus for supplying a sample having a solvent discharge side flow path for supplying a sample solution containing a sample and a solvent to the solvent discharge side flow path The sample solution in the discharge side flow path for sample solution is supplied to the discharge side flow path switching means capable of switching between the discharge path, the sample solution discharge side flow path, and the solvent discharge side flow path. And a high-pressure pump for the sample solution for performing the operation, and a device that is a device in which a predetermined amount of the sample solution is intermittently supplied to the mobile phase flow path by switching the flow path by the discharge-side flow path switching means. .

  In the apparatus, the sample solution is supplied to the liquefied gas or the supercritical fluid by means configured in the same manner as the means for supplying the solvent to the mobile phase flow path. The mobile phase to be generated is stably generated in the mobile phase channel, and the pressure fluctuation of the mobile phase accompanying the supply of the sample is prevented. In addition, since the solvent and the sample solution are respectively supplied to the liquefied gas or the supercritical fluid by different supply means configured similarly to the mobile phase flow path, the flow path for supplying the solvent and the pump Contamination by the sample is prevented.

According to the present invention, in the supercritical fluid chromatography including the step of mixing the liquefied gas or the supercritical fluid and the solvent in the mobile phase flow path whose pressure is adjusted, the solvent mixed in the mobile phase flow path Instead, a predetermined amount of sample solution containing a sample and a solvent is supplied to the mobile phase flow path, so in the supercritical fluid chromatography device, the sample is supplied to the mobile phase with a simple configuration without pressure fluctuations. can do.

  Further, according to the present invention, in the supercritical fluid chromatography apparatus having the mobile phase generating means for generating the mobile phase containing the supercritical fluid and the solvent in the mobile phase flow path whose pressure is adjusted by the pressure adjusting means. The supercritical fluid has means for supplying a predetermined amount of the sample solution to the solvent suction side channel for supplying the solvent to the solvent high pressure pump for supplying the solvent to the liquefied gas or supercritical fluid. In a chromatography apparatus, a sample can be supplied to a mobile phase with a simple configuration without pressure fluctuation.

  Further, according to the present invention, in the supercritical fluid chromatography apparatus having the mobile phase generating means for generating the mobile phase containing the supercritical fluid and the solvent in the mobile phase flow path whose pressure is adjusted by the pressure adjusting means. A solvent discharge side channel and a sample solution discharge from a solvent high pressure pump for supplying a solvent to a liquefied gas or supercritical fluid to a solvent discharge side channel for supplying solvent to a mobile phase channel Supercritical because it has a discharge-side flow path switching means for switching between the side flow paths and a sample solution high-pressure pump for supplying the sample solution in the discharge-side flow path for the sample solution to the discharge-side flow path switching means In a fluid chromatography device, the sample can be supplied to the mobile phase with a simple configuration without pressure fluctuations, and contamination of the solvent flow path due to the sample supply is prevented. Door can be.

  In the present invention, the target substance is an optical isomer, and a column containing a separation agent for separating the target substance from a sample is used, and the separation agent contains an optically active polysaccharide or polysaccharide derivative. The use of the agent is more effective from the viewpoint of increasing the separation efficiency of the target substance and suppressing the diffusion of the sample in the mobile phase flow path. The separation agent is cellulose, amylose, their ester derivatives, or their carbamates. The use of a derivative is even more effective.

<Sample supply method>
The present invention relates to a sample supplied to a mobile phase containing a supercritical fluid and a solvent and separates the target substance from the sample containing the target substance. It is a method of supplying.

  The supercritical fluid chromatography includes a step of mixing a liquefied gas or a supercritical fluid to constitute a supercritical fluid and a solvent in a mobile phase flow path in which the pressure is adjusted. The supercritical fluid chromatography is not particularly limited as long as it is a method including the mixing step. In addition, the supercritical fluid chromatography can further include other steps than the mixing step as long as the effects of the present invention are not impaired.

  In the mixing step, the liquefied gas or the supercritical fluid for forming the supercritical fluid and the solvent are mixed in the mobile phase flow path whose pressure is adjusted. The pressure of the mobile phase flow path is not particularly limited as long as it is sufficient to form a supercritical state of the supercritical fluid. The pressure in the mobile phase channel can be adjusted by known pressure adjusting means such as a back pressure adjusting valve.

  The liquefied gas for constituting the supercritical fluid is not particularly limited as long as it is a liquefied gas that becomes a supercritical fluid by at least one of pressurization and heating. The liquefied gas is appropriately selected according to the type of supercritical fluid to be configured. Examples of the means for pressurizing the liquefied gas include a high-pressure pump that is usually used in a high-performance liquid chromatography apparatus. Examples of means for heating the liquefied gas include a heat exchanger for heating the liquefied gas. Can be mentioned.

The supercritical fluid is a substance that is in one or both of a state above a critical pressure and a point above a critical temperature (ie, a supercritical state). Examples of substances used as the supercritical fluid include carbon dioxide, ammonia, sulfur dioxide, hydrogen halide, nitrous oxide, hydrogen sulfide, methane, ethane, propane, butane, ethylene, propylene, halogenated hydrocarbons, water, and the like. Can be mentioned.

  As the solvent, one kind or two or more kinds are selected from various known solvents according to the kind of the target substance, the kind of the supercritical fluid, and the like. Examples of the solvent include methanol, lower alcohols such as ethanol and 2-propanol, ketones such as acetone, acetonitrile, ethyl acetate, and water.

  The method for mixing the liquefied gas or supercritical fluid and the solvent is not particularly limited as long as the method can be mixed in the mobile phase flow path. As such a method, for example, there is a method in which a liquefied gas or a supercritical fluid is quantitatively supplied to the mobile phase flow path with a high-pressure pump, while a solvent is quantitatively supplied to the mobile phase flow path with a high-pressure pump. Can be mentioned.

  The mixing ratio of the liquefied gas or supercritical fluid and the solvent is the type of the target substance, the type of column used to separate the target substance, the type of separating agent contained in the column, the type of supercritical fluid, and the solvent. The ratio of the solvent with respect to the entire mobile phase is 5 to 30% by volume, so that the target substance is separated with high separation efficiency in supercritical fluid chromatography. It is preferable from the viewpoint. The ratio of the solvent in the mobile phase is determined based on the supply amount of the liquefied gas or supercritical fluid to the mobile phase flow path and the supply amount of the solvent to the mobile phase flow path. Can be adjusted by relatively controlling by.

  In the present invention, instead of the solvent mixed with the liquefied gas or the supercritical fluid in the mobile phase channel, a predetermined amount of the sample solution containing the sample and the solvent is intermittently supplied to the mobile phase channel. The method for intermittently supplying the sample solution is not particularly limited as long as the sample is intermittently present in the solvent supplied to the mobile phase flow path. Examples of such a method include a method of supplying a predetermined amount of sample to the solvent, and a method of switching between the supply of the solvent and the supply of the sample solution to the mobile phase flow path.

  The method of supplying a predetermined amount of sample to the solvent can be performed using a known injector (injector) used in, for example, high performance liquid chromatography. The method of switching between the supply of the solvent and the supply of the sample solution to the mobile phase flow path is, for example, a means capable of opening and closing the flow path such as a two-way valve or a three-way valve, or a liquid feeding means such as the high-pressure pump. Can be used.

  The sample solution is not particularly limited as long as it is a solution containing a sample and a solvent. The solvent in the sample solution becomes one component that forms the mobile phase when supplied to the mobile phase flow path. Therefore, if the amount of the solvent in the sample solution is too small, the separation of the target substance may be hindered due to a change in the composition of the mobile phase. Efficiency may be reduced. From such a viewpoint, the content of the sample in the sample solution is preferably 0.2 to 15% by mass, and more preferably 0.5 to 10% by mass with respect to the entire sample solution.

  Further, the sample solution may contain components other than the sample and the solvent as long as they do not adversely affect the separation of the target substance or adversely affect the change in the composition of the mobile phase. Such other components can be selected from, for example, known organic solvents.

The amount of the sample solution supplied intermittently instead of the solvent is not particularly limited as long as it is an appropriate amount for separating the target substance. If the amount of the sample solution supplied intermittently is too large, the sample may spread over a wide range in the mobile phase flow path, and the target substance may not be sufficiently separated. If the amount of the sample solution supplied intermittently is too small, the separation efficiency of the target substance may be lowered.
From such a viewpoint, it is preferable that the amount of the sample solution supplied intermittently is 0.1 to 1200 mL with respect to the amount of the solvent supplied to the mobile phase flow path. The amount of the sample solution supplied intermittently can be adjusted by the injection amount of the sample, the time for switching to the supply of the sample solution, the operation of the high-pressure pump, and the like.

  The present invention employs various methods for suppressing a decrease in the separation efficiency of the target substance when the sample diffuses in the mobile phase channel, and for suppressing the diffusion of the sample in the mobile phase channel. It is preferable. For example, as a method for suppressing a decrease in separation efficiency due to sample diffusion, for example, a method using a column having a high resolution as a column for separation can be mentioned, and as a method for suppressing sample diffusion, a flow path for mobile phase is used. A method of increasing the flow rate of the mobile phase in is used.

  The column used in supercritical fluid chromatography is appropriately selected according to the type of the target substance. For example, when the target substance is an optical isomer, a column containing an optically active polysaccharide or polysaccharide derivative as a separating agent can be used as a column having a high resolution of the target substance.

  The polysaccharide may be any of a synthetic polysaccharide, a natural polysaccharide, and a natural product-modified polysaccharide as long as it is an optically active polysaccharide. The polysaccharide is preferably a polysaccharide with a high regularity of the binding mode, and is preferably a chain polysaccharide. Examples of the polysaccharide include various polysaccharides, and cellulose and amylose are particularly preferable.

  The polysaccharide derivative is not particularly limited as long as it is a polysaccharide derivative that can be used for separation of optical isomers. As such a polysaccharide derivative, for example, a polysaccharide derivative containing the polysaccharide as a skeleton, wherein at least a part of the hydroxyl group and amino group of the polysaccharide is substituted with a functional group that acts on an optical isomer in a sample. Can be mentioned. Examples of the polysaccharide derivative include various polysaccharide derivatives, and at least one selected from an ester derivative of cellulose, a carbamate derivative of cellulose, an ester derivative of amylose, and a carbamate derivative of amylose is particularly preferable. Specific examples of the polysaccharide derivative include various polysaccharide derivatives described in International Publication No. 95/23125 pamphlet.

  The separating agent may be formed into particles, or may be supported on a particulate carrier and filled in the column, or may be accommodated in the column while being supported on an integral carrier accommodated in the column. Alternatively, it may be accommodated in the column as an integral molded product made of a separating agent.

  For example, as described in Japanese Patent No. 2783819, a particulate separation agent packed in a column is prepared by dissolving the polysaccharide or polysaccharide derivative as a separation agent in a solvent, and using the resulting solution as water or the like. The insoluble solvent in which the separating agent does not dissolve, preferably in the insoluble solvent containing a dispersing agent such as an anionic surfactant, can be produced by dropping the insoluble solvent with stirring. The separating agent obtained as described above can be used in the present invention.

  A packing material composed of a particulate carrier packed in a column and a separating agent supported on the column can be produced, for example, by supporting the separating agent on the carrier by physical adsorption or chemical adsorption. The separation agent is supported on the carrier by immersing the carrier in a solution containing the separation agent and, if necessary, another compound, reacting with the other compound as necessary, and distilling off the solvent in the solution, It can be carried out by a method of replacing the solvent in the solution with another solvent. The filler thus obtained can be used in the present invention.

Examples of the carrier include porous organic carriers such as polystyrene, polyacrylamide, polyacrylate, and derivatives thereof; porous materials such as silica, alumina, magnesia, glass, kaolin, titanium oxide, silicate, and hydroxyapatite. Inorganic carrier; and the like.

  A column in which a separating agent is supported on an integrated carrier accommodated in the column is accommodated in the column, or the integral carrier accommodated in the column is filled with the solution of the separating agent, The solvent can be produced by distilling off the solvent from the monolithic carrier filled with the solution or by replacing the solvent from the monolithic carrier filled with the solution with another solvent. The column thus obtained can be used in the present invention.

  Examples of the integrated carrier accommodated in the column include, for example, a porous shaped body described in JP 2000-515627 A, an integrated adsorbent described in JP 2002-505005, A known inorganic carrier such as an inorganic porous column described in JP-A-6-265534 or an improved product thereof can be used.

  As an integrated carrier accommodated in the column, that is, a column in which the integral carrier is accommodated, for example, Chromolith (registered trademark of MERCK) can be mentioned.

  The separating agent formed into an integral molded product accommodated in the column can be produced by forming a porous body having a predetermined shape with the polysaccharide or the polysaccharide derivative. As such a production method, for example, the insoluble solvent, bubbles, or polygen is dispersed in the solution of the separating agent, the dispersed solution is stored in a column tube, and the solvent is distilled off from the solution stored in the column tube. Alternatively, a method may be used in which the polygen is dissolved by passing through a column tube a cleaning solvent that replaces and dissolves the polygen if necessary. As the polygen, known particles such as resins and salts that are excellent in solubility in the washing solvent for the molded product are used. The separating agent thus obtained can be used in the present invention.

  It is preferable to use a column filled with particles consisting only of a separating agent or a column containing an integrated molded article consisting only of a separating agent from the viewpoint of enhancing the separation ability of optical isomers. In addition, the use of a column containing an integrated carrier carrying a separating agent or a column containing an integrated molded article made only of a separating agent suppresses pressure loss due to the column and reduces the mobile phase. This is preferable from the viewpoint of increasing the flow rate of the mobile phase in the working channel.

  The flow rate of the mobile phase in the mobile phase channel can be appropriately set according to the type of column used. If the flow rate of the mobile phase in the mobile phase flow path is too high, the target substance may not be sufficiently separated by the column. If the flow rate of the mobile phase in the mobile phase flow path is too slow, the target substance may diffuse into the mobile phase flow path, and the target substance may not be sufficiently separated by the column. From such a viewpoint, the flow rate of the mobile phase in the mobile phase flow path varies depending on the scale of the supercritical fluid chromatography apparatus, but is preferably 10 mL / min to 120 L / min. The flow rate of the mobile phase in the mobile phase flow path is adjusted by the supply speed of the liquefied gas or supercritical fluid and solvent to the mobile phase flow path, the inner diameter of the tube constituting the mobile phase flow path, the type of column, etc. It is possible.

<Sample supply device>
A sample supply device of the present invention is a sample supplied to a mobile phase containing a supercritical fluid and a solvent, and is a supercritical fluid chromatography device for separating a target substance from a sample containing the target substance The apparatus for supplying a sample to the mobile phase.

The supercritical fluid chromatography apparatus includes a mobile phase channel, a pressure adjusting unit for adjusting the pressure of the mobile phase channel, and a mobile phase channel whose pressure is adjusted by the pressure adjusting unit. The apparatus is not particularly limited as long as the apparatus includes a mobile phase generation unit that generates a mobile phase containing a critical fluid and a solvent.

  As the mobile phase flow path, a normal pressure-resistant tube used for the mobile phase flow path in high performance liquid chromatography or supercritical fluid chromatography can be used. As the pressure adjusting means, a known means such as a back pressure adjusting valve can be used as described in the sample supply method.

  The mobile phase generation means supplies a supercritical fluid high-pressure pump for supplying a liquefied gas or supercritical fluid for forming a supercritical fluid to the mobile phase flow channel, and a solvent to the mobile phase flow channel. A solvent high-pressure pump and a solvent suction-side flow path for supplying the solvent to the solvent high-pressure pump. Alternatively, the mobile phase generation means has a solvent discharge side flow path for supplying the solvent discharged from the solvent high pressure pump to the mobile phase flow path instead of or in addition to the solvent suction side flow path. .

  The high-pressure pump for supercritical fluid and the high-pressure pump for solvent are not particularly limited as long as the fluid can be sent to the mobile phase flow path in which the pressure at which the supercritical fluid can exist is adjusted. As these high-pressure pumps, as described above in the sample supply method, high-pressure pumps that are usually used in high-performance liquid chromatography apparatuses can be used.

  The solvent suction side flow path is not particularly limited as long as it is a flow path capable of supplying a normal pressure solvent to the solvent high pressure pump. A stainless steel pressure-resistant tube or a flexible tube made of a resin such as a fluororesin can be used for such a channel. The solvent discharge-side flow path can be configured in the same manner as the mobile phase flow path.

  The mobile phase generation means may further include another device for generating a mobile phase. Examples of such other devices include a device for pressurizing a mixed fluid containing a liquefied gas and a solvent, and a device for heating the mixed fluid. For example, a known device such as a high pressure pump or a back pressure adjusting valve can be used as the device for pressurization, and a known device such as a heat exchanger can be used as the device for heating.

  The supercritical fluid chromatography apparatus is provided with other components that are usually used to separate a target substance in a sample. Examples of such other components include a column for separating a target substance in a sample, a detector for detecting a component in a mobile phase after passing through the column, and a target separated by a column. Examples thereof include a fraction collector for taking out a target substance from a mobile phase containing the substance and whose pressure adjustment by the pressure adjusting means is released.

  As the column, it is preferable to use a column having a high ability to separate the target substance from the viewpoint of suppressing a decrease in the separation efficiency of the target substance when the sample diffuses in the mobile phase flow path. When the target substance is an optical isomer, a column containing the separating agent or packing material described above in the sample supply method is preferably used as the column.

  In the sample supply device of the present invention, either the first sample supply device or the second sample supply device is used depending on the configuration of the mobile phase generating means.

  The first sample supply device is a device that can be used when the mobile phase generating means has the solvent suction side flow path, and a sample solution containing a sample and a solvent is supplied to the solvent suction side flow. Means for supplying to the road. The solvent and the sample solution are the same as the solvent and the sample solution in the sample supply method described above.

  The means for supplying the sample solution may be a means for supplying the sample solution to the solvent suction side flow path, or a sample is supplied to the solvent suction side flow path to which the solvent is supplied to suck the solvent. It may be a means for supplying the sample solution to the side channel.

  Among the means for supplying the sample solution, the means for supplying the sample solution to the solvent suction side channel supplies, for example, a sample solution tank for storing the sample solution and a sample solution stored in the sample solution tank. And a suction side switching means that can switch between the sample solution suction side channel and the solvent suction side channel.

  The sample solution suction side channel is not particularly limited as long as it is capable of supplying a normal pressure sample solution to the solvent suction side channel. As such a channel, a channel similar to the solvent suction side channel can be used.

  The suction side switching means is not particularly limited as long as it can switch between the solvent suction side flow path and the sample solution suction side flow path with respect to the solvent high-pressure pump. Examples of such means include a two-way valve provided in each flow path extending from a connection point between the sample solution suction side flow path and the solvent suction side flow path, and a three-way valve provided at the connection point. Is mentioned.

  Among the means for supplying the sample solution, the means for supplying the sample to the solvent suction side channel includes, for example, an injector used in a high performance liquid chromatography apparatus.

  The first sample supply device can appropriately supply two types of fluids with a single high-pressure pump, and can generate a mobile phase containing a sample with a simple configuration.

  The second sample supply device is a device that can be used when the mobile phase generation means has the solvent discharge side flow path, and is a device that supplies a predetermined amount of the sample solution to the mobile phase flow path. It is. The second sample supply device is capable of switching between a discharge side channel for sample solution for supplying the sample solution, a discharge side channel for sample solution, and a discharge side channel for solvent. And a sample solution high-pressure pump for supplying the sample solution in the sample solution discharge-side flow path to the discharge-side flow path switching means.

  A normal pressure-resistant tube can be used for the sample solution discharge-side flow channel in the same manner as the mobile phase flow channel. Further, the discharge side flow path switching means is provided at the connection point of each flow path to be switched or a two-way valve provided in each flow path to be switched, similarly to the suction side flow path switching means. A three-way valve or the like can be used. As the high-pressure pump for the sample solution, a high-pressure pump usually used in a high-performance liquid chromatography apparatus can be used as in the high-pressure pump for supercritical fluid and the high-pressure pump for solvent.

  The second sample supply device may have other devices than the above-described components. As such other devices, for example, a sample solution bypass pipe connecting the discharge side and the suction side of the sample solution high-pressure pump in the sample solution discharge side flow path, and the sample solution bypass pipe can be freely opened and closed. And a two-way valve. According to such a configuration, the sample solution discharge-side channel is depressurized between the sample solution high-pressure pump and the discharge-side channel switching means, and the amount of sample solution supplied to the sample solution discharge-side channel Can be adjusted.

In the case of using the second sample supply device, the mobile phase generation means includes a solvent bypass pipe connecting the discharge side and the suction side of the solvent high-pressure pump in the solvent discharge side flow path, and the solvent bypass. You may further have a two-way valve which can open and close a pipe | tube. According to such a configuration, it is possible to depressurize the solvent discharge side flow path between the solvent high pressure pump and the discharge side flow path switching means and to adjust the amount of solvent supplied to the sample solution discharge side flow path. It becomes possible.

  The second sample supply device generates a mobile phase containing a sample with a simple configuration in which a sample solution flow path provided with a high-pressure pump is connected to a solvent flow path via a flow path switching means. Is possible. In addition, the second sample supply device can prevent contamination of the solvent high-pressure pump and the solvent flow path by the sample.

  Operations such as switching of a flow path by a valve, opening and closing of a valve, and operation of a high-pressure pump in the sample supply apparatus of the present invention may be performed by, for example, a worker of a supercritical fluid chromatography apparatus or a supercritical fluid chromatography. It may be performed by a control device for instructing and controlling various operations in the graphic apparatus.

  Hereinafter, embodiments of the present invention will be described more specifically.

<First embodiment>
As shown in FIG. 1, the supercritical fluid chromatography apparatus used in the present embodiment includes a cylinder 1 filled with high-pressure carbon dioxide, and a liquefied gas by cooling the carbon dioxide supplied from the cylinder 1. A heat exchanger 2, a high-pressure pump 3 that quantitatively pumps the liquefied gas generated in the heat exchanger 2, a solvent tank 4 that contains a solvent, and a solvent tank 4 that is supplied to the liquefied gas sent by the high-pressure pump 3. And a heat exchanger 6 for heating the fluid mixture of the liquefied gas and the solvent to convert the liquefied gas in the fluid mixture into a supercritical fluid.

  The supercritical fluid chromatography apparatus includes a sample solution tank 7 for storing a sample solution containing a sample containing a target substance (for example, an optical isomer) and a solvent, and a flow path on the suction side of the high-pressure pump 5. A three-way valve 8 for switching between a flow path to the solvent tank 4 and a flow path to the sample solution tank is provided.

  The supercritical fluid chromatography apparatus includes a column 9 for separating a target substance in a sample supplied to the mobile phase, a detector 10 for detecting the substance in the mobile phase passing through the column 9, a high pressure And a back pressure valve 11 for keeping the pressure in the system from the pump 3 to the detector 10 at a predetermined pressure.

  Further, the supercritical fluid chromatography device contains a plurality of gas-liquid separation devices 12 for gas-liquid separation of the mobile phase that has passed through the back pressure valve 11, and a plurality of liquid phases separated by each gas-liquid separation device 12. The first tank 13 and a plurality of second tanks 14 connected to the respective first tanks 13.

  Further, the supercritical fluid chromatography apparatus includes a gas recovery pipe 15 that connects each gas-liquid separation apparatus 12 and the heat exchanger 2, and a gas-liquid separation apparatus for separating a liquid from the recovery gas flowing through the gas recovery pipe 15. 16, a third tank 17 containing the liquid phase separated by the gas-liquid separator 16, and a fourth tank 18 connected to the third tank 17.

Furthermore, the supercritical fluid chromatography apparatus includes a regulator 19 for supplying carbon dioxide gas from the cylinder 1 at a predetermined pressure, and a check valve for preventing a backflow of gas from the heat exchanger 2 to the cylinder 1. 20, a buffer tank 21 for storing the liquefied gas generated in the heat exchanger 2, a two-way valve 22 provided between the back pressure valve 11 and each gas-liquid separator 12, the first tank 13, and the first tank 13 A two-way valve 23 provided between the second tank 14, a check valve 26 for preventing a backflow of gas to each gas-liquid separator 12, a third tank 17 and a fourth tank 18. And a two-way valve 27 provided between the two.

  Each device from the high pressure pump 3 to the back pressure valve 11 is connected by a pressure resistant tube (for example, a stainless steel tube). The discharge side of the high pressure pump 5 is connected to a pipe a connecting the high pressure pump 3 and the heat exchanger 6 by a pressure resistant pipe b. The suction side of the high-pressure pump 5, the three-way valve 8, the three-way valve 8 and the solvent tank 4, and the three-way valve 8 and the sample solution tank 7 are respectively connected by ordinary tubes (for example, fluororesin flexible tubes) c to e. It is connected.

  Each component between the high pressure pump 3 and the back pressure valve 11 corresponds to the mobile phase flow path. The back pressure valve 11 corresponds to the pressure adjusting means. The high pressure pump 3 corresponds to the supercritical fluid high pressure pump, the high pressure pump 5 corresponds to the solvent high pressure pump, the tubes c and d correspond to the solvent suction side flow path, and the heat exchanger 6 Corresponds to the mobile phase generating means. The tube e corresponds to the sample solution suction side flow path, the three-way valve 8 corresponds to the suction side flow path switching means, and these correspond to the sample supply apparatus of the present invention (the first sample supply apparatus). .

  As the gas-liquid separators 12 and 16, for example, the gas-liquid separator shown in FIGS. 2 and 3 is used. This gas-liquid separator is formed along a cylindrical outer cylinder 31 having both ends open, a flange portion 32 that closes an opening at one end of the outer cylinder 31, and a tangent to the inner peripheral surface of the cross-sectional shape of the outer cylinder 31. And an inner cylinder that is open at the outer cylinder 31 and has both ends open, and extends through the flange 32 to the other end side of the outer cylinder 31 from the introduction section 33. 34, a jacket 35 that covers the outer peripheral wall surface of the outer cylinder 31 and forms a circulation path for the heat medium, and a mobile phase supply pipe 36 that is connected to the introduction portion 33 through the jacket 35.

  The jacket 35 is provided with a heat medium supply port 37 on the other end side and a heat medium discharge port 38 on one end side. The mobile phase supply pipe 36 passes through the jacket 35 from one end portion of the jacket 35, exits from the other end side of the jacket 35, and is connected to the introduction portion 33. The mobile phase supply pipe 36 is spirally wound around the outer periphery of the outer cylinder 31 in the jacket 35.

  The mobile phase supply pipe 36 is connected to the two-way valve 22 (the check valve 26 in the gas-liquid separator 16). The inner cylinder 34 is connected to the check valve 26 (the gas recovery pipe 15 connected to the heat exchanger 2 in the gas-liquid separator 16). The other end of the outer cylinder 31 is connected to the first tank 13 (the third tank 17 in the gas-liquid separator 16). The supply port 37 and the discharge port 38 are connected to a heat medium circulating means (not shown).

  Note that the outer cylinder 31 has an elliptical or multi-walled inner peripheral wall shape for the purpose of causing a bias in the flow velocity of the mobile phase introduced into the outer cylinder 31 or applying an appropriate impact. A non-circular cylinder such as a square may be used.

  Next, separation of the target substance by the supercritical fluid chromatography apparatus will be described.

  First, carbon dioxide is supplied from the cylinder 1 to the heat exchanger 2 at an appropriate initial pressure. The carbon dioxide cooled by the heat exchanger 2 becomes liquefied gas and is stored in the buffer tank 21. The liquefied gas stored in the buffer tank 21 is quantitatively pumped by the high-pressure pump 3 and pressurized to a predetermined pressure (for example, critical pressure) defined by the back pressure valve 11, while the heat exchanger is passed through the pipe a. 6 is supplied.

  In the case of generating only the mobile phase, the three-way valve 8 connects the pipe c and the pipe d, and the solvent (for example, methanol) is supplied from the solvent tank 5 to the high-pressure pump 5 through the pipe d and the pipe c. Is done. The solvent supplied to the high-pressure pump 5 is quantitatively pumped, supplied to the pipe a through the pipe b, and merged with the liquefied gas in the pipe a and mixed. The fluid mixture of the liquefied gas and the solvent is supplied to the heat exchanger 6 and heated to a predetermined temperature (for example, a critical temperature or a set temperature of the column 9). By this heating, the liquefied gas in the mixed fluid becomes a supercritical fluid, and a mobile phase containing the supercritical fluid and the solvent is generated.

  In the case of supplying the sample to the mobile phase, the three-way valve 8 is operated to connect the tube c and the tube e. A sample solution (for example, a racemic methanol solution) is supplied from the sample solution tank 7 to the high-pressure pump 5 through the pipe e and the pipe c. The sample solution supplied to the high-pressure pump 5 is quantitatively pumped, supplied to the pipe a through the pipe b, and merged with the liquefied gas in the pipe a and mixed. The sample mixed fluid containing the liquefied gas, the solvent, and the sample is supplied to the heat exchanger 6 and heated to the predetermined temperature. By this heating, the liquefied gas in the sample mixed fluid becomes a supercritical fluid, and the mobile phase containing the sample is generated.

  When a predetermined time elapses after the pipe c and the pipe e are connected by the three-way valve 8, the three-way valve 8 is operated to connect the pipe c and the pipe d again. By operating the three-way valve 8, the liquid supplied to the high-pressure pump 5 is switched from the sample solution to the solvent. In this way, a predetermined amount of the sample solution is intermittently supplied to the tube a instead of the solvent mixed with the liquefied gas in the tube a.

  The mobile phase produced by containing a predetermined amount of the sample solution is composed of a column 9 (for example, a unitary molded product of silica) containing a separating agent (for example, a polysaccharide derivative) corresponding to the target substance, and the carrier. The polysaccharide derivative supported thereon is sent to a column) housed in a column tube. In the column 9, the target substance is separated from the sample. The target substance is detected by the detector 10.

  When the target substance is detected by the detector 10, for example, a detection signal from the detector 10 is transmitted to a control device (not shown), and the control device opens any one of the two-way valves 22 and the other two-way valve. Each two-way valve 22 is operated so that 22 is closed. In this way, the supply of the mobile phase to the gas-liquid separator 12 is switched according to the target substance.

  On the other hand, the mobile phase containing the target substance is sent to the back pressure valve 11. The mobile phase that has passed through the back pressure valve 11 is released from the pressure adjustment by the back pressure valve 11, the supercritical state of carbon dioxide in the mobile phase is released, and is supplied to a predetermined gas-liquid separator 12.

  The mobile phase sent to the gas-liquid separator 12 is gas-liquid separated. Carbon dioxide forming the supercritical fluid is separated as a gas phase, and the target substance and solvent are separated as a liquid phase. The liquid phase is stored in the first tank 13 and further stored in the low pressure second tank 14.

  The liquid phase accommodated in the second tank 14 is taken out from the second tank 14, and the solvent and the target substance are separated from the liquid phase by a known method such as vacuum concentration or vacuum distillation. Is obtained. The solvent may be purified as necessary and reused in the mobile phase. The carbon dioxide separated by the gas-liquid separator 12 is sent to the gas-liquid separator 16.

  When another target substance is detected by the detector 9, the two-way valve 22 corresponding to the gas-liquid separator 12 is closed. Then, the two-way valve 22 corresponding to the other gas-liquid separator 12 is opened, and the gas-liquid separation of the mobile phase containing the other target substance is similarly performed by the other gas-liquid separator 12.

Carbon dioxide (recovered gas) sent to the gas-liquid separator 16 is gas-liquid separated by the gas-liquid separator 16. A small amount of liquid phase (solvent) contained in the recovered gas is stored in the third tank 17 and then stored in the fourth tank 18 and discarded.

  The recovered gas purified by the gas-liquid separator 16 is sent to the heat exchanger 2 through the gas recovery pipe 15. When the pressure of the recovered gas is higher than the initial pressure defined by the regulator 19, the recovered gas is supplied to the heat exchanger 2 and liquefied. When the pressure of the recovered gas is lower than the initial pressure defined by the regulator 19, new carbon dioxide gas from the cylinder 1 is supplied to the heat exchanger 2 and liquefied.

  In the present embodiment, the operation of the three-way valve 8 may be performed by an operator. However, for example, the control device described above connects the pipes c and d with a preset operation time (for example, the three-way valve 8). The three-way valve 8 may be operated according to the time during which the pipes c and e are connected).

  In the present embodiment, the supercritical fluid chromatography apparatus includes components from the high pressure pump 3 to the back pressure valve 11, the high pressure pump 5, the pipes c to e, the three-way valve 8, and the heat exchanger 6. Since the sample solution is supplied to the mobile phase flow path in the same manner as the supplied solvent for generating the mobile phase, the sample solution is supplied to the mobile phase flow path without causing pressure fluctuation. And a mobile phase containing a predetermined amount of sample can be generated without causing pressure fluctuations.

  In the present embodiment, the supply of the sample solution described above can be realized with a simple configuration of the three-way valve 8, the pipe e, and the sample solution tank 7.

  In the present embodiment, since the supercritical fluid chromatography device further includes the second tank 14 from the gas-liquid separation device 12, the target substance is taken out from the mobile phase from which the supercritical state of the supercritical fluid is released. be able to. Therefore, it can be used for production of a target substance such as an optical isomer. In the production of the target substance, there is no pressure fluctuation associated with the supply of the sample, so that the target substance can be produced stably.

  In the present embodiment, since the supercritical fluid chromatography apparatus further includes the fourth tank 18 from the gas recovery pipe 15, the carbon dioxide is recovered from the mobile phase from which the supercritical state of the supercritical fluid is released. In addition, the recovered carbon dioxide can be reused to produce a supercritical fluid in the mobile phase. Therefore, the amount of new carbon dioxide used can be reduced, and the cost required for carbon dioxide, which is a raw material for the supercritical fluid, can be suppressed.

  In the present embodiment, a gas recovery pipe 15 that supplies recovered gas and a cylinder 1 that supplies new gas are connected to the heat exchanger 2 that generates liquefied gas. The cylinder 1 includes a regulator 19 and a check valve 20. Since it is connected to the heat exchanger 2 via the recovered gas, the recovered gas can be reused in preference to the new gas according to the pressure of the recovered gas. Therefore, the cost required for carbon dioxide can be further suppressed, which is more effective from the viewpoint of increasing the productivity of the target substance.

  In the present embodiment, an example in which a column having a low pressure loss is used for the column 9 has been shown. However, by using such a column 9, the flow rate of the mobile phase can be easily increased. The diffusion of the sample in the tube up to the column 9 is suppressed, and the target substance can be separated with high separation efficiency.

In addition, instead of the column 9 with low pressure loss, a column filled with particles formed with a separating agent such as a polysaccharide derivative, or a column containing an integral molded product formed with the separating agent is used. When a column having a high separation ability for the substance is used, even if the sample diffuses in the tube from the heat exchanger 6 to the column 9, the high separation ability of the column and the decrease in separation efficiency due to the diffusion of the sample are offset, Again, the target substance can be separated with high separation efficiency.

<Second Embodiment>
As shown in FIG. 4, the supercritical fluid chromatography apparatus used in the present embodiment is used in the first embodiment except that the connection form of the solvent tank 4 and the sample solution tank 7 to the tube a is different. It is configured in the same manner as the supercritical fluid chromatography apparatus.

  In the supercritical fluid chromatographic apparatus used in the present embodiment, a pressure-resistant tube f is connected to a pressure-resistant tube a that connects the high-pressure pump 3 and the heat exchanger 6, and the tube f is connected to the tube f. A three-way valve 41 is connected. Pressure-resistant pipes g and i are further connected to the three-way valve 41, the pipe g is connected to the discharge side of the high-pressure pump 5, and the pipe i is connected to the discharge side of the high-pressure pump 42. . The suction side of the high-pressure pump 5 is connected to the solvent tank 4 by a normal pipe h, and the suction side of the high-pressure pump 42 is connected to the sample solution tank 7 by a normal pipe j.

  The pipes f and g correspond to the solvent discharge side flow path, and the high pressure pumps 3 and 5 and the heat exchanger 6 correspond to the mobile phase generating means. The pipes i and j correspond to the discharge side flow path for the sample solution, the three-way valve 41 corresponds to the discharge side flow path switching means, and the high pressure pump 42 corresponds to the high pressure pump for the sample solution. It corresponds to a sample supply device (the second sample supply device).

  In the present embodiment, the generation of the liquefied gas of carbon dioxide and the supply of the generated liquefied gas to the pipe a are performed in the same manner as in the first embodiment.

  In the present embodiment, the three-way valve 41 and the high-pressure pumps 5 and 41 are interlocked, and a pipe-side high-pressure pump that is opened by switching the three-way valve 41 is started and closed by the three-way valve 41. The high-pressure pump on the side is stopped. The linkage between the three-way valve 41 and the high-pressure pumps 5 and 41 is performed by, for example, the control device described above.

  In the case of generating only the mobile phase, the three-way valve 41 connects the pipe f and the pipe g, and the solvent (for example, methanol) is supplied from the solvent tank 5 to the high-pressure pump 5 through the pipe h. The solvent supplied to the high-pressure pump 5 is quantitatively pumped, supplied to the pipe a through the pipes g and h, and merged with the liquefied gas in the pipe a and mixed.

  When supplying the sample to the mobile phase, the three-way valve 41 is operated to connect the tube f and the tube i. By operating the three-way valve 41, the high-pressure pump 5 is stopped and the high-pressure pump 42 is started. The sample solution is supplied from the sample solution tank 7 to the high-pressure pump 42 through the pipe j. The sample solution supplied to the high-pressure pump 42 is quantitatively pumped, supplied to the pipe a through the pipe i and the pipe f, and merges with the liquefied gas in the pipe a and mixed.

  When a predetermined time elapses after the pipe f and the pipe i are connected by the three-way valve 8, the three-way valve 8 is operated to connect the pipe f and the pipe g again. By operating the three-way valve 8, the high pressure pump 42 is stopped, the high pressure pump 5 is started, and only the solvent is again supplied to the pipe a by the high pressure pump 5. In this way, a predetermined amount of the sample solution is intermittently supplied to the tube a instead of the solvent mixed with the liquefied gas in the tube a.

  For various operations such as generation of mobile phase after supply of solvent or sample solution to tube a or generation of mobile phase containing sample, separation of target substance, fractionation, and gas recovery, the first implementation This is performed in the same manner as in the above.

  In the present embodiment, the supercritical fluid chromatography apparatus includes components from the high pressure pump 3 to the back pressure valve 11, the high pressure pump 5, the pipes f, g, and i, the three-way valve 41, and the high pressure pump 42. And the heat exchanger 6, and the sample solution is intermittently supplied to the mobile phase flow path by switching the flow path by the three-way valve 41, so that the sample solution is used for the mobile phase without causing pressure fluctuations. A mobile phase that can be supplied to the flow path and contains a predetermined amount of sample can be generated without causing pressure fluctuations.

  In the present embodiment, the above-described supply of the sample solution can be realized with a simple configuration including the three-way valve 41, the pipe i, the high-pressure pump 42, the pipe j, and the sample solution tank 7.

  In the present embodiment, the same effects as those of the first embodiment are achieved with respect to stable production of the target substance, reduction of the cost required for carbon dioxide, and separation of the target substance with high separation efficiency.

<Third embodiment>
As shown in FIG. 5, the supercritical fluid chromatography apparatus used in the present embodiment includes a bypass pipe that bypasses the high-pressure pump and connects the suction side and the discharge side of the high-pressure pump, a high-pressure pump for solvent, and a sample. Except for the point provided with respect to the high-pressure pump for solution, it is comprised similarly to the supercritical fluid chromatography apparatus used in 2nd embodiment.

  That is, in the supercritical fluid chromatography apparatus used in the present embodiment, the solvent bypass pipe 51 that bypasses the high-pressure pump 5 and connects the pipe g and the pipe h, and the solvent bypass pipe 51 for opening and closing. The two-way valve 53, the sample solution bypass pipe 52 that bypasses the high-pressure pump 42 and connects the pipe i and the pipe j, and the two-way valve 54 that opens and closes the sample solution bypass pipe are shown in FIG. Further provided for the supercritical fluid chromatography apparatus shown.

  In the case of such a configuration, instead of starting / stopping the high-pressure pump 5 associated with the switching of the three-way valve 41, or in addition to starting / stopping the high-pressure pump 5, the two-way valve is opened / closed, thereby Can be removed. In this embodiment, after generation of the liquefied gas of carbon dioxide, supply of the generated liquefied gas to the tube a, supply of the solvent or sample solution to the tube a, supply of the solvent or sample solution to the tube a For the various operations such as the generation of the mobile phase or the generation of the mobile phase containing the sample, the separation of the target substance, fractionation, and the recovery of the gas, except for the operation of the two-way valves 53 and 54, the second This is performed in the same manner as the embodiment.

  Regarding the operation of the two-way valve, specifically, when only the mobile phase is generated, when the three-way valve 41 is operated and the pipe g is closed, the two-way valve 53 is opened and the high-pressure pump 5 is stopped. When the pipe g is closed, the solvent pumped by the high-pressure pump 5 is confined in the pipe g, but the internal pressure of the pipe g is maintained at a normal pressure by opening the two-way valve 53.

  Similarly, when supplying the sample to the mobile phase, when the three-way valve 41 is operated to close the pipe i, the two-way valve 54 is opened and the high-pressure pump 42 is stopped. When the pipe i is closed, the solvent pumped by the high-pressure pump 42 is confined in the pipe i, but the internal pressure of the pipe i is maintained at normal pressure by opening the two-way valve 54.

  When the pipe g and the pipe f are connected by operating the three-way valve 41, the two-way valve 53 is closed. When the pipe i and the pipe f are connected by operating the three-way valve 41, the two-way valve 54 is closed. It is done.

In the present embodiment, the supply amount of the solvent to the pipe a by the high-pressure pump 5 may be adjusted by opening the two-way valve 53 at an appropriate opening degree when the pipe g and the pipe f are connected. . Similarly, the supply amount of the sample solution to the pipe a by the high-pressure pump 42 may be adjusted by opening the two-way valve 54 at an appropriate opening when the pipe i and the pipe f are connected.

  The opening and closing of the two-way valves 53 and 54 described above can be controlled by, for example, the control device described above.

  In the present embodiment, sealing of the pressurized solvent or sample solution in the tube is prevented. Therefore, in this embodiment, it is possible to prevent an unexpected pressure fluctuation accompanying the supply of the sample solution or the solvent, such as a pressure fluctuation to the high pressure side. Further, in this embodiment, for example, the exhaustion of the seal of the joint of the pipe is suppressed, which is more effective from the viewpoint of maintenance management of the supercritical fluid chromatography apparatus. In addition to these effects, the present embodiment has the effects of the second embodiment.

It is a figure which shows roughly the structure of the supercritical fluid chromatography apparatus used for one embodiment of this invention. It is sectional drawing which shows the longitudinal cross-section of the principal part of an example of the gas-liquid separator used by this Embodiment. It is sectional drawing which shows the cross section of the principal part when the gas-liquid separator shown in FIG. 2 is cut | disconnected by the AA line. It is a figure which shows schematically the structure of the supercritical fluid chromatography apparatus used for other embodiment of this invention. It is a figure which shows schematically the structure of the principal part in other embodiment of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cylinder 2, 6 Heat exchanger 3, 5, 42 High pressure pump 4 Solvent tank 7 Sample solution tank 8, 41 Three-way valve 9 Column 10 Detector 11 Back pressure valve 12 Gas-liquid separator 13 First tank 14 Second tank DESCRIPTION OF SYMBOLS 15 Gas collection pipe 17 3rd tank 18 4th tank 19 Regulator 20, 26 Check valve 21 Buffer tank 22, 23, 27, 53, 54 Two-way valve 31 Outer cylinder 32 Flange part 33 Introduction part 34 Inner cylinder 35 Jacket 36 Mobile phase supply pipe 37 Heat medium supply port 38 Heat medium discharge port 51 Solvent bypass pipe 52 Sample solution bypass pipe a to j pipe

Claims (8)

A method of supplying a sample to a mobile phase in supercritical fluid chromatography, wherein the sample is supplied to a mobile phase containing a supercritical fluid and a solvent, and the target material is separated from a sample containing the target material. Because
The supercritical fluid chromatography includes a step of mixing a liquefied gas or a supercritical fluid for constituting a supercritical fluid and the solvent in a mobile phase flow path whose pressure is adjusted,
In the method of supplying the sample, a predetermined amount of sample solution containing the sample and the solvent is used for the mobile phase instead of the solvent mixed with the liquefied gas or the supercritical fluid in the mobile phase channel. A method characterized by intermittently supplying the flow path.   The supercritical fluid chromatography further includes a step of separating the target substance from the sample in the mobile phase by a column for separating the target substance, and the target substance is an optical isomer, The method according to claim 1, wherein a column containing a separating agent for separating optical isomers is used, and a separating agent containing an optically active polysaccharide or polysaccharide derivative is used as the separating agent. Cellulose or amylose is used as the polysaccharide, and at least one selected from cellulose ester derivatives, cellulose carbamate derivatives, amylose ester derivatives, and amylose carbamate derivatives is used as the polysaccharide derivative. Item 3. The method according to Item 2.
In a supercritical fluid chromatography apparatus for separating a target substance from a sample supplied to a mobile phase containing a supercritical fluid and a solvent and containing the target substance, the sample is added to the mobile phase. A device for supplying,
The supercritical fluid chromatography apparatus comprises: a mobile phase flow path; a pressure adjusting means for adjusting the pressure of the mobile phase flow path; and the mobile phase flow whose pressure is adjusted by the pressure adjusting means. Mobile phase generation means for generating a mobile phase containing a supercritical fluid and a solvent in the path,
The mobile phase generation means includes a high-pressure pump for supercritical fluid for supplying a liquefied gas or supercritical fluid for constituting a supercritical fluid to the mobile phase flow channel, and the solvent for the mobile phase flow channel. A solvent high-pressure pump for supplying to the solvent, and a solvent suction-side flow path for supplying the solvent to the solvent high-pressure pump,
The apparatus for supplying a sample has means for supplying a predetermined amount of a sample solution containing the sample and the solvent to the solvent suction side flow path.   The means for supplying the sample solution includes a sample solution tank for storing the sample solution and a sample solution suction side flow for supplying the sample solution stored in the sample solution tank to the solvent suction side flow path. 5. The apparatus according to claim 4, further comprising: a channel, and suction side channel switching means capable of switching between the sample solution suction side channel and the solvent suction side channel. In a supercritical fluid chromatography apparatus for separating a target substance from a sample supplied to a mobile phase containing a supercritical fluid and a solvent and containing the target substance, the sample is added to the mobile phase. A device for supplying,
The supercritical fluid chromatography apparatus comprises: a mobile phase flow path; a pressure adjusting means for adjusting the pressure of the mobile phase flow path; and the mobile phase flow whose pressure is adjusted by the pressure adjusting means. Mobile phase generation means for generating a mobile phase containing a supercritical fluid and a solvent in the path,
The mobile phase generation means includes a high-pressure pump for supercritical fluid for supplying a liquefied gas or supercritical fluid for constituting a supercritical fluid to the mobile phase flow channel, and the solvent for the mobile phase flow channel. A solvent high pressure pump for supplying to the solvent, and a solvent discharge side flow path for supplying the solvent discharged from the solvent high pressure pump to the mobile phase flow path,
The apparatus for supplying the sample comprises: a sample solution discharge side channel for supplying a sample solution containing the sample and the solvent to the solvent discharge side channel; and the sample solution discharge side flow A discharge side flow path switching means capable of switching between a path and the solvent discharge side flow path, and a sample solution for supplying the sample solution in the discharge side flow path for the sample solution to the discharge side flow path switching means A device having a high-pressure pump, wherein a predetermined amount of sample solution is intermittently supplied to the mobile phase flow channel by switching the flow channel by the discharge side flow channel switching means.   The target substance is an optical isomer, and the supercritical fluid chromatography apparatus uses a column containing a separating agent for separating optical isomers in a mobile phase stream downstream of the mobile phase generating means. The apparatus according to any one of claims 4 to 6, further comprising a path, wherein the separating agent is a separating agent containing an optically active polysaccharide or polysaccharide derivative. The polysaccharide is cellulose or amylose, a polysaccharide derivative is an ester derivative of cellulose, a carbamate derivative of cellulose, an ester derivative of amylose and, claim 7, wherein at least either selected from a carbamate derivative of amylose The device described.

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