Detection system and method for detecting chiral compounds

Abstract

A detection system and a method for detecting chiral compounds, wherein the multi-mode analytical system is suitable for separating non-chiral and chiral components and for resolving and detecting enantiomers, and the analytical system includes an injection system, a liquid chromatography system as a first dimension chromatograph, a supercritical chromatography system as a second dimension chromatograph, a trap system, a connection system, and a detector, the liquid chromatography system includes a reversed-phase liquid column, the supercritical chromatography system includes one or more chiral columns, the connection system includes a first multi-port valve and a second multi-port valve, and the detector is a mass spectrometry detector.

Description

[Technical Field]

[0001] The present invention belongs to the field of chemical detection, and specifically relates to a detection and analysis system for chiral compounds, which is applicable to the rapid and automatic separation of non-chiral and chiral components, as well as the efficient resolution of enantiomers, and a method for detecting chiral chemicals using the detection and analysis system. [Background technology]

[0002] Chiral compounds are a unique structural phenomenon in small molecules. Due to the presence of chiral centers (chiral atoms), small molecules have the ability to exhibit different spatial isomers. Furthermore, due to differences in spatial structure, even if they have the same molecular formula, chemical structure, or atomic composition, they can exhibit completely different properties and activities.

[0003] Chiral compounds typically have one or more chiral centers (chiral atoms), and when synthesized or extracted, various isomers may exist. These isomers may be diastereomers or enantiomers. Therefore, from a research perspective, it is necessary to investigate the differences in properties and activities between different isomers, which requires the separation of single-structure chiral compounds for separate study. On the other hand, separation is also necessary to obtain chiral compounds with specific activities and specific spatial configurations. Furthermore, as research deepens, the needs for these two aspects are increasing.

[0004] Chiral drugs have generally been found to have different biological activities and pharmacological effects in the body. For example, the right-handed form of thalidomide has sedative properties, while the left-handed form can cause fetal malformations. Therefore, in the field of drug synthesis, it is particularly important to study the chiral structures of synthesized compounds and the differences between different chiral structures. Furthermore, in terms of the synthesis of chiral compounds, especially pharmaceutical chiral compounds, it is also desirable to obtain chiral compounds with specific spatial structures by using various catalysts and other control measures.

[0005] Chromatography is also widely used for the separation, qualitative analysis, and quantitative analysis of compounds. Chromatography has also been used to separate and analyze non-chiral and chiral components in fields such as drug synthesis. Chromatography techniques commonly used to resolve chiral drugs include gas chromatography, high-performance liquid chromatography (HPLC), and supercritical fluid chromatography (SFC). SFC, in particular, uses a supercritical fluid as the mobile phase and a chiral column. Due to the different solubilities and mechanisms of action of the chiral components in the mobile and stationary phases, SFC has been successful in separating chiral drug enantiomers with high resolution and good selectivity.

[0006] Furthermore, when performing chiral resolution, one-dimensional chromatography is currently mainly applied to the analysis of single components. Non-chiral components are generally separated and detected by liquid chromatography (LC), while chiral components are generally separated and detected by normal-phase chromatography.

[0007] Cited Document 1 reports a method for separating tetrahydropalmatine using high-performance liquid chromatography (HPLC), in which chiral resolution of tetrahydropalmatine is performed using an α-acid glycoprotein (AGP) chiral column and a cellulose tris(4-methylbenzoate) chiral column, based on the difference in chiral recognition due to π-π interactions and dipole-dipole interactions between THP enantiomers and the chiral stationary phase.

[0008] However, it is usually difficult to separate the non-chiral and chiral components in a chiral compound sample containing complex components using only LC, etc. Therefore, conventional one-dimensional chromatography does not provide sufficient separation, and currently, general one-dimensional chromatographic systems are no longer able to meet the detection needs of complex samples.

[0009] In addition, attempts have been made to combine multiple dimensional chromatography, i.e., multidimensional chromatography technology, from the perspectives of improving separation efficiency, convenience of detection, and accuracy. Multidimensional chromatography technology has been found to play an important role in separating complex components. In particular, multidimensional chromatographic systems combining SFC have attracted increasing attention due to the special selectivity, efficient separation capabilities, energy saving, and environmental protection characteristics of supercritical fluid separation technology.

[0010] Among current multidimensional chromatographic systems, two-dimensional (2D) chromatography is one of the most widely used multidimensional chromatographic techniques. Two-dimensional chromatography involves connecting two independent chromatographic systems in series, allowing components that could not be successfully separated in the first chromatographic system to be further separated in the second chromatographic system. Compared to first-dimensional chromatography, two-dimensional chromatography can provide larger peak capacity and better resolution, selectivity, and sensitivity. Currently, two-dimensional chromatographic systems are widely used in fields such as pharmaceutical analysis and food analysis.

[0011] In Reference 2, the heart-cutting, two-dimensional liquid-phase, chiral resolution method was used to separate and analyze the enantiomers of tetrahydropalmatine in Yuanhu analgesic tablets.

[0012] The conversion between the two separation modes and the transfer of separated components in two-dimensional chromatography rely on the valve interface switching technology, and the loss of first-dimensional separation power can be reduced by designing an appropriate interface. Multidimensional chromatographic systems based on valve switching technology are playing a larger role.

[0013] Cited Document 3 discloses a method for separating and analyzing complex chiral compounds using RPLC-SFC, in which the analytes are eluted or passed through a mobile phase in the first dimension of RPLC, and a UV detector is used in combination to provide separation results for non-chiral and chiral components, the analytes are focused using a trap column, and then the components are injected into the second dimension of SFC to perform chiral separation, and the separation results of the chiral enantiomers are detected using MS.

[0014] Although the above-mentioned research has been conducted on the separation of non-chiral and chiral components, and enantiomers, it cannot be said that the results are sufficient in terms of improving the detection efficiency of separation and resolution, convenience and accuracy of detection, and there is room for further improvement. [Prior art document]

[0015] References: Reference 1: "Separation of Tetrahydroberberine Enantiomers by High-Performance Liquid Chromatography Using Chiral Stationary Phases," Hong Zhanying et al., "Chinese Journal of Pharmacopoeia," 2006, 03, 223-226 Reference 2: "Quality Analysis of Yuanhu Analgesic Tablets by Fingerprinting and Enantiomer Ratio Methods," Liu Juanru et al., Journal of Drug Analysis, 2022, 42(08), 1472-1481 Cited document 3: CN107073358B Summary of the Invention [Problem to be solved by the invention]

[0016] As mentioned above, analytical methods using one-dimensional or multidimensional chromatographic systems have been attempted for chiral analysis of target substances or separation and detection of non-chiral and chiral components. Among these, two-dimensional chromatographic detection methods have demonstrated their advantages, allowing for smooth separation of non-chiral and chiral components and separation and detection of enantiomers. For example, in Reference 3, for compounds with multiple chiral centers, an independent first detector (e.g., a spectrophotometric detector) is used as a monitor, and different eluates from the first-dimension chromatograph are collected in different containers of a trap (each container collects a pair of enantiomers). Further, chiral separation is performed by a second-dimension chromatograph, and detection is performed by an independent second detector (e.g., a mass spectrometer).

[0017] Although documents such as Reference 3 can be applied to the separation and detection of enantiomers of compounds with multiple chiral centers, the analytical efficiency is not sufficient, and the detection system is not simple. In particular, whether separating and detecting enantiomers of compounds with one chiral center or multiple chiral centers, an independent first detector must be used to determine the retention time of specific eluting components.

[0018] Furthermore, there is currently a significant demand for catalyst screening for large-scale and rapid detection of enantiomers, and the product components obtained by each catalyst can be simple (e.g., containing mainly one pair of enantiomer products) or complex (containing multiple pairs of enantiomers). However, the use of a detection system such as that in Reference 3 cannot meet the needs of the industry, especially when detecting products with only one pair of enantiomers, or when the target detectable product is only one pair of enantiomers, the design of the detection system in Reference 3 is too complicated. Currently, other conventional technologies do not provide a simpler and faster detection method.

[0019] Therefore, the first object of the present invention is to provide a two-dimensional detection analysis system that has a simpler design and is more efficient in separating non-chiral and chiral components and separating enantiomers, and that includes a liquid chromatography system as the first dimension chromatograph and a supercritical fluid chromatography system as the second dimension chromatograph, and that, by designing the connected system, allows detection to be completed using only one mass spectrometric detector. Overall, the present invention not only satisfies the separation of non-chiral and chiral components and the effective resolution of one or more pairs of enantiomers, but also significantly improves the efficiency of resolution detection, especially when there is only one pair of enantiomers or when the target analyte is only one pair of enantiomers.

[0020] Furthermore, the present invention also provides a method for detecting separation of non-chiral and chiral components and resolution of enantiomers using the above detection system, which exhibits excellent detection efficiency, particularly in the resolution detection of enantiomers having one chiral center or a pair of enantiomers.

[0021] Another object of the present invention is to provide a method for screening catalysts that can efficiently evaluate the selectivity of catalysts with respect to chiral properties using the above-mentioned detection method of the present invention.

[0022] Furthermore, the present invention also provides a method for evaluating the quality of herbal medicines using the analytical system of the present invention. [Means for solving the problem]

[0023] As a result of long-term research, the inventors of the present invention have found that the above-mentioned problems can be solved by implementing the following invention.

[0024] [1] First, the present invention provides a multi-mode analytical system for use in detecting chiral compounds, comprising: The analytical system includes an injection system, a liquid chromatography system as a first-dimension chromatograph, a supercritical chromatography system as a second-dimension chromatograph, a trap system, a connection system, and a detector; the liquid chromatographic system includes a reversed-phase liquid column, the supercritical chromatographic system includes one or more chiral columns, the connection system includes a first multi-port valve and a second multi-port valve, and the detector is a mass spectrometry detector; The first multi-way valve is configured to: i) providing a connection between said liquid chromatographic system and said trapping system; or ii) providing a connection between said trapping system and said supercritical chromatography system; The second multi-way valve is configured to switch the valve position so that the mass spectrometric detector: i') detecting the eluate from said liquid chromatographic system; or ii') Providing an analytical system adapted to detect effluent from said supercritical chromatograph. [2] The analytical system according to [1], wherein the injection system is an automatic injection system and is in communication with the reversed-phase liquid column. [3]. The analytical system according to [1] or [2], wherein the liquid chromatograph system further comprises a liquid chromatography mobile phase supply device, and the reversed-phase liquid column is a silica gel column modified with a hydrophobic group. [4] The analytical system according to any one of [1] to [3], wherein the trap system is a trap ring or a trap having one or more storage portions. [5] The analytical system according to any one of [1] to [4], wherein the supercritical chromatography system includes a supercritical chromatography mobile phase supply device and a back pressure control device. [6] The analytical system according to any one of [1] to [5], wherein the supercritical chromatography system includes two or more chiral columns. [7] The analytical system according to any one of [1] to [6], wherein the first multi-port valve and the second multi-port valve are both six-port valves. [8]. A first working path in which the reversed-phase liquid column communicates with the trap system via the first multi-way valve, and the trap system further communicates with the detector via the second multi-way valve; The analytical system according to any one of [1] to [7], further comprising: a second operating path in which the trapping system is connected to the supercritical chromatography system via the first multi-way valve, and the chiral column further communicates with the detector via the second multi-way valve. [9]. The analytical system described in [8], wherein the connection in the second operating path connects a mobile phase supply device of a supercritical chromatography system with the trap system via the first multi-way valve.

[10] The analysis system according to [8] or [9], further comprising an operating path that uses the first operating path as a backup operating path.

[11] The analytical system according to any one of [1] to

[10] , wherein the first and second multi-way valves of the connection system switch positions using a self-control system.

[12] . The analytical system according to any one of [1] to

[11] , wherein the mass spectrometry detector is configured to display or respond only to the characteristic mass spectrometry signal of the target chiral compound.

[13] . A method for detecting a chiral compound, comprising: A detection method in which detection is performed using the analytical system according to any one of [1] to

[12] , wherein the chiral compound has one or more chiral centers.

[14] The detection method according to

[13] , wherein the chiral compound is a low molecular weight compound.

[15] .a. When the test sample only has one pair of enantiomer compounds or the target detection product in the test sample is only one pair of enantiomers, after the pair of enantiomers are collected in the trap system according to the corresponding retention time, or after the corresponding mass spectrometry peak detection signal of the compound appears in the detector, switch the first multi-way valve and the second multi-way valve to enter the second operation mode; b. If the test sample contains two or more pairs of enantiomer compounds, collect each pair of enantiomers into the trap system according to the corresponding retention time, and after the collection is completed, switch the first and second multi-way valves to enter the second operation mode; a first operation mode in which the reversed-phase liquid column is connected to the trapping system via the first multi-way valve, and the trapping system is further connected to the detector via the second multi-way valve, and one or more pairs of enantiomers in a test sample are sequentially collected in the trapping system; The detection method according to

[13] or

[14] , further comprising the second operating mode in which the trapping system is connected to the supercritical chromatography system via the first multi-port valve, and the chiral column is further connected to the detector via the second multi-port valve, and the peak intensity or area of ​​a characteristic mass spectrometry peak for each pair of enantiomers eluted from the chiral column is further detected by the detector.

[16] . The detection method described in

[15] , wherein the retention times of the enantiomers under condition a and the retention times of the multiple pairs of enantiomers under condition b are determined by a preliminary operation mode, in which the reversed-phase liquid column is connected to the trap system via the first multi-way valve, and the trap system is further connected to the detector via the second multi-way valve.

[17] . The detection method according to any one of

[13] to

[16] , wherein the detector detects the eluate from the chiral column and determines the ratio of the peak intensities or areas of the characteristic mass spectrometry peaks of two isomers of one or more pairs of enantiomers, thereby determining the content ratio of the two isomers.

[18] The present invention also provides a method for evaluating a catalyst for synthesizing a chiral compound, comprising: The present invention also provides an evaluation method for evaluating the corresponding catalyst by synthesizing the chiral compound using each catalyst to be evaluated, detecting each product obtained from each catalyst using the detection method described in any one of

[13] to

[17] , and measuring the peak intensity or area ratio of the two characteristic mass spectrometry peaks of a pair of target enantiomers.

[19] . The present invention further provides a method for synthesizing a drug containing a chiral compound, wherein the synthesis of the chiral compound includes screening a catalyst using the evaluation method described in

[16] .

[20] Furthermore, the present invention provides a method for evaluating the quality of herbal medicines, which comprises analyzing the chiral components in the herbal medicines using the detection method described in any one of

[13] to

[17] and measuring the relative proportions of the enantiomers, thereby evaluating the quality of the herbal medicines. [Effects of the Invention]

[0025] By implementing the above invention, the present invention can achieve the following effects. 1) The detection and analysis system and detection method provided by the present invention can separate non-chiral and chiral compounds in complex composition systems, efficiently separate enantiomers, and detect and analyze their relative contents. 2) The detection analysis system and detection method provided by the present invention can efficiently separate and detect enantiomers of chiral compounds having one or more chiral centers, especially chiral compounds having one chiral center, or when the target detectant is only one pair of enantiomers, and exhibits very good detection efficiency. 3) Because of its extremely high detection efficiency, it can provide an effective evaluation method for large-scale and efficient screening of catalysts in drug synthesis. 4) Compared with conventional two-dimensional chiral detection systems, the present invention can complete the detection process with only one mass spectrometric detector, which simplifies the device configuration and improves economy. [Brief explanation of the drawings]

[0026] [Figure 1] Physical diagram of the analytical system of the present invention [Figure 2] Schematic diagram of the analytical system of the present invention (first operating path / first operating mode) [Figure 3] Schematic diagram of the analytical system of the present invention (second operating path / second operating mode): 10: reversed-phase liquid chromatography; 11: liquid phase pump (supplying mobile phase A); 12: liquid phase pump (supplying mobile phase B); V1 / V2: six-way valve; 20: group of chiral columns (6 columns); 21: supercritical fluid supply device (supplying mobile phase C); 22: liquid phase pump (supplying mobile phase D); 23: backpressure control device; 30: trap ring; 40: mass spectrometry detector [Figure 4] Chromatogram of a mixed solution of L-THP and D-THP reference materials analyzed with an LC-SFC multidimensional chromatographic system [Figure 5] Chromatogram of Yuanhu analgesic tablets analyzed by LC-SFC multidimensional chromatographic system DETAILED DESCRIPTION OF THE INVENTION

[0027] The present invention will be described in detail below. The technical features described below will be explained based on representative embodiments and specific examples of the present invention, but the present invention is not limited to these embodiments and specific examples. Unless otherwise defined, the following applies.

[0028] As used herein, a numerical range expressed as "numerical value A to numerical value B" means a range including the limit values ​​A and B. As used herein, "basic" or "substantial" means within a 5%, preferably 3%, and more preferably 1% standard deviation from the theoretical model or data. As used herein, "plurality" refers to a number greater than or equal to two. As used herein, "%" refers to volume percentage, or "V%." As used herein, the terms "communication" or "connection" mean that a plurality of devices or members can form a flow path for the mobile phase and / or the test component. In this specification, the terms "chromatography column" and "column" have the same meaning. In this specification, the term "switching" refers to switching the position or state of a multi-way valve to block a path that was originally connected and to connect a device or component that was originally disconnected. In this specification, the term "may" includes both cases where some processing is performed and cases where some processing is not performed. As used herein, "normal temperature" or "room temperature" refers to an indoor temperature of 23±2°C. As used herein, "optional" or "optional" means that the event or circumstance described below may or may not occur, and the description includes both cases where the event occurs and cases where it does not occur. As used herein, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "embodiments," etc., mean that the particular element (e.g., a feature, structure, property, and / or characteristic) described in connection with that embodiment is included in at least one embodiment described herein and may or may not be present in other embodiments. Furthermore, it should be understood that such elements may be combined in any manner in various embodiments.

[0029] The present invention provides a method for separating non-chiral and chiral components of a target substance having a complex composition, and for resolving and detecting the enantiomers in the chiral component, and a corresponding analytical system, which is particularly suitable for resolving and detecting the enantiomers of a low molecular weight compound having one chiral center. In the present invention, by connecting a liquid chromatography system as a first-dimension chromatograph, a supercritical chromatography system as a second-dimension chromatograph, a trap system, and a detector via a connection system, different operating paths or operating modes can be formed to perform the above detection simply and efficiently.

[0030] <First aspect> In a first aspect of the present invention, there is provided a multi-mode analytical system for use in detecting chiral compounds, the analytical system comprising an injection system, a liquid chromatography system as a first dimension chromatograph, a supercritical chromatography system as a second dimension chromatograph, a trap system, a connection system, and a detector.

[0031] (Chiral compounds and detection targets) The chiral compounds described in the present invention are low molecular weight compounds, and mainly refer to non-polymeric single molecular compounds, which are distinguished from so-called "oligomers" and "polymers."

[0032] In some specific embodiments of the present invention, chiral compounds may have one or more chiral centers or chiral atoms, such as those having 1 to 5 chiral centers, preferably 1 to 3 chiral centers, more preferably 1 or 2 chiral centers, and most preferably 1 chiral center.

[0033] A compound with one chiral center may have one pair of enantiomers, and a compound with multiple (e.g., n) chiral centers may have two n It may have multiple diastereomers and enantiomers, and may also have multiple pairs of enantiomers.

[0034] Chiral compounds have different properties depending on their spatial structure, and different spatial structures often show obvious differences in drug activity and properties. Therefore, studying compounds with different spatial structures is particularly important in drug screening, which has increased the need for enantiomer resolution.

[0035] In principle, there are no special requirements for the detection target of the present invention, and it may be, for example, any composition containing the chiral compound of the present invention. In some preferred embodiments of the present invention, the detection target may be a finished drug, for example, a solid, liquid, suspension, or slurry-based drug, and may be subjected to any necessary pretreatment to purify or concentrate the test component using a method such as organic solvent or water extraction before detection. In other preferred embodiments of the present invention, the detection target may be a (direct) reaction product obtained at the end of a synthetic reaction of a pharmaceutical molecule compound, or a purified product thereof purified by any method.

[0036] In some specific embodiments of the present invention, the target substance to be detected may contain a non-chiral component and a chiral component, or may contain only a chiral component having at least one pair of enantiomers.

[0037] (Injection System) The injection system in the analytical system of the present invention is not particularly limited, but a manual sample injector or an automatic sample injector can be used, and preferably an automatic sample injector can be used.

[0038] (1st dimension chromatograph) The first-dimensional chromatograph in the analytical system of the present invention is a liquid chromatography system, more specifically, a reversed-phase liquid chromatography system, and is preferably a high-speed reversed-phase liquid chromatography system or an ultra-high-speed reversed-phase liquid chromatography system, such as the ultra-high-performance liquid chromatography system Nexera LC-40 manufactured by Shimadzu Corporation.

[0039] In the present invention, a reversed-phase liquid chromatographic system comprises one or more reversed-phase liquid columns, which in some embodiments are placed in a column oven during use.

[0040] The stationary phase in the reversed-phase liquid chromatographic system of the present invention may be silica gel modified with a hydrophobic group, and the hydrophobic group may be various hydrocarbon groups such as a C8 group, a C18 group, or a phenyl group. In some preferred embodiments of the present invention, silica gel modified with C18 is used as the stationary phase.

[0041] The mobile phase in the reversed phase liquid chromatography system of the present invention can generally include an aqueous phase (hereinafter simply referred to as "mobile phase A") and an organic phase (hereinafter simply referred to as "mobile phase B").

[0042] In some embodiments, mobile phase A may be an aqueous solution containing a good volatility buffer salt, such as an aqueous solution of highly volatile ammonium bicarbonate.

[0043] The mobile phase B can be selected from organic phases. The organic phase can be a nitrile-based substance or an alcohol-based substance, and typically, acetonitrile can be used as the organic phase.

[0044] Furthermore, the liquid chromatographic system of the present invention can further include a mobile phase supply device for supplying the mobile phase to the liquid chromatographic system. In some specific embodiments, a (liquid phase) pump is used as the mobile phase supply device. In some preferred embodiments, two (liquid phase) pumps are used to supply the mobile phase A and the mobile phase B, respectively.

[0045] (trap system) The trapping system of the present invention functions as a device for capturing eluted components (chiral components) from the first-dimensional chromatograph, and then separates the enantiomers using the second-dimensional chromatograph described below.

[0046] In some specific embodiments, the trapping system of the present invention may be a trap having one or more eluted component storage portions, and in other specific embodiments, may be a single trap ring.

[0047] The trap is suitable for collecting multiple analytes, for example, when the analyte has multiple chiral compounds with different molecular formulas, or when the analyte has multiple chiral centers but the same molecular formula, etc. In this case, the trap can be programmed to collect different eluting components in different receptacles at different times.

[0048] The trap ring is applicable to chiral compounds that contain only one pair of enantiomers with the same molecular formula in the analyte, or when the target substance in the detection subject contains only one pair of enantiomers.

[0049] In some preferred embodiments of the present invention, a trap ring is used as the trapping system.

[0050] (2nd dimension chromatograph) The second dimensional chromatograph in the analytical system of the present invention is a supercritical chromatography system.

[0051] Supercritical fluid chromatography (SFC, hereinafter also referred to as "supercritical chromatography") is chromatography that uses a supercritical fluid as the mobile phase.

[0052] Supercritical fluid chromatography has the properties of both gas chromatography and liquid chromatography, making it possible to analyze samples with high boiling points or low volatility that are not suitable for gas chromatography. It also has faster analytical speeds and better conditions than high-performance liquid chromatography.

[0053] In the present invention, one or more chiral chromatography columns (multiple columns used in parallel) can be used as the chromatography column in the supercritical chromatography system. There is no particular requirement for the number of chiral chromatography columns, and the number of chiral chromatography columns contributes to improved separation ability and (screening) efficiency. In some preferred embodiments of the present invention, two or more or three or more, for example, five to seven, chiral chromatography columns can be used. Examples of chiral chromatography include silica gel-based polar group-modified chromatography columns, cellulose-modified chromatography columns, and starch-modified chromatography columns. These columns are generally commercially available. Examples include Opti-Chiral® A1, A2, A3, A4, and A5 (amylose series), Opti-Chiral® C1, C2, C3, C4, C5, and C6 (cellulose series), CHIRALPAK® brand polysaccharide derivative-coated chiral columns for SFC, and polysaccharide derivative solvent-resistant chiral columns for SFC (bonded chiral columns). In some embodiments of the invention, the chiral column is disposed entirely within a column oven when in use.

[0054] Furthermore, the mobile phase in the supercritical chromatography of the present invention includes a supercritical fluid (mobile phase C) and a modifier fluid (mobile phase D). The mobile phase formed from both can be supplied via a mobile phase supply device.

[0055] The supercritical fluid refers to a state of matter intermediate between gas and liquid under conditions above the critical temperature and above the critical pressure. Suitable supercritical fluids may include supercritical carbon dioxide or supercritical ethane. In some embodiments of the present invention, the supercritical fluid of the present invention is selected from supercritical carbon dioxide. The operating temperature and pressure are primarily determined by the type of supercritical fluid selected.

[0056] In the present invention, when supercritical carbon dioxide is used as mobile phase C, the operating temperature is 31°C or higher, preferably 35°C or higher, and the operating pressure is 7.3 MPa or higher, preferably 7.5 MPa or higher. In some preferred embodiments, the operating temperature may be 40 to 60°C, and the operating pressure is preferably 7.5 to 15 MPa, and typically the operating temperature may be 50°C, and the pressure may be 10 MPa.

[0057] In the present invention, the modifier fluid used as the mobile phase D can be used as a substance for adjusting the polarity of the supercritical fluid.

[0058] The modifier fluid can be selected from an aqueous solution of an alcohol-based or nitrile-based substance, and a pH adjusting component can be used as an auxiliary agent if necessary. Examples of alcohol-based substances include various aliphatic alcohols such as methanol and isopropanol. Examples of nitrile-based substances include acetonitrile.

[0059] Furthermore, the mobile phase C of the supercritical chromatography of the present invention can be supplied by an independent supercritical fluid supply device, and the mobile phase D can be supplied to the chromatography column by a (liquid phase) pump. In some embodiments, the supercritical fluid column of the present invention can be placed in a column oven. Also, as shown in FIG. 2, a pressure control unit (e.g., a backpressure control unit BPR) is also provided along with the supercritical fluid column. In some embodiments, a pressure control unit is placed at the end of this supercritical fluid chromatography section to remove the supercritical fluid.

[0060] As a supercritical chromatograph that can be used in the present invention, for example, a commercially available product such as "Nexera UC" manufactured by Shimadzu Corporation (Kyoto, Japan) is available.

[0061] (Connection System) The connection system of the present invention includes at least two multi-port valves, and switching the position of the multi-port valves allows switching between the first and second dimension chromatographs, or more specifically, switching at least two multi-port valves allows different operating paths or modes of operation between the first and second dimension chromatographs, as described below.

[0062] In some preferred embodiments of the present invention, the connection system includes two multi-way valves, a first multi-way valve and a second multi-way valve; The first multi-way valve is configured to: i) providing a connection between said liquid chromatographic system and said trapping system; or ii) providing a connection between said trapping system and said supercritical chromatography system; The second multi-way valve is configured to switch the valve position so that the mass spectrometric detector: i') detecting the eluate from said liquid chromatographic system; or ii') detecting effluent from said supercritical chromatographic system; "Or" here means switching the position of the multi-way valve to transition from one operating state to another.

[0063] In addition, in some specific embodiments, the multi-port valve used in the present invention may be a six-port valve or a ten-port valve, and preferably, the first multi-port valve and the second multi-port valve are both six-port valves.

[0064] Furthermore, when a plurality of chiral columns are used in a supercritical chromatography system, it is preferable to connect two or more multi-way valves to realize a screening function for different chiral columns.

[0065] (detector) The detector of the present invention is a mass spectrometric detector, and the above-mentioned analytical system of the present invention can achieve all desired functions using only one mass spectrometric detector by switching the operating path or operating mode. In other words, the analytical system of the present invention makes it possible to use only one mass spectrometric detector without using more mass spectrometric detectors or other types of detectors (e.g., UV-visible type detectors).

[0066] Furthermore, in some preferred embodiments of the present invention, the mass spectrometric detector can be further configured to reflect or respond only to a characteristic mass spectrometric peak of the detected compound, which may be the peak of the most intense ion fragment of the detected compound, or one to three peaks of the most intense ion fragments.

[0067] Furthermore, the mass spectrometer detector may be a triple quadrupole mass spectrometer, a single quadrupole mass spectrometer, a high-resolution mass spectrometer Q-TOF, etc. These detectors are commercially available, for example, as LCMS-8050.

[0068] (Other devices or systems) In addition to the various systems and devices described above, the analytical system of the present invention may optionally use other auxiliary devices and systems, such as, but not limited to, a temperature control device, a guard column device for liquid chromatography, and an automatic control device, as long as they do not affect the manifestation of the effects of the present invention.

[0069] <Second aspect> In a second aspect of the present invention, there is provided a method for separating a non-chiral component from a chiral component, and for resolving and detecting enantiomers in a chiral component, using the analytical system of the first aspect above.

[0070] Specifically, the present invention provides different actuation paths and, in turn, different actuation modes by switching the position of the connection system.

[0071] Specifically, the analysis system of the present invention comprises: a first working path in which the reversed-phase liquid column communicates with the trapping system via the first multi-way valve, and the trapping system further communicates with the detector via the second multi-way valve; The trap system is connected to the supercritical fluid chromatography system via the first multi-port valve, and the chiral column operates via a second operating path in which the trap system is connected to the supercritical fluid chromatography system via the second multi-port valve. Furthermore, the term "connected" refers to the communication between the trap system and a mobile phase supplier of the supercritical fluid chromatography system via the first multi-port valve.

[0072] Optionally, the analytical system may also provide a backup working path, ie, use the first working path as the backup working path.

[0073] Therefore, the above different working paths can complete the detection through the following working states or working modes during the detection. In a first operation mode, the reversed-phase liquid column is connected to the trapping system through the first multi-port valve, and the trapping system is further connected to the detector through the second multi-port valve, and one or more pairs of enantiomers in a test sample are sequentially collected in the trapping system under the following conditions: a. If the test sample contains only a pair of enantiomer compounds as the analyte, after the pair of enantiomers is collected in the trap system according to the corresponding retention time, or after the corresponding characteristic mass spectrometry peak detection signal of the compound appears in the detector, the first multi-way valve and the second multi-way valve are switched to switch to the second operating mode. Preferably, in this case, the trap system uses a trap ring. Furthermore, the test sample containing only a pair of enantiomer compounds as the analyte means that the test sample contains only a pair of enantiomers, or the target analyte in the test sample is only a pair of enantiomers. b. When the test sample contains two or more pairs of enantiomers, each pair of enantiomers is collected in the trap system according to its corresponding retention time, and after the collection is completed, the first and second multi-way valves are switched to switch to the second operation mode. Preferably, under this condition, the trap system can use a trap with multiple storage compartments. In the second operating mode, the trapping system is connected to the supercritical chromatography system via the first multi-way valve, and the chiral column is further connected to the detector via the second multi-way valve, and the intensity or area of ​​the characteristic mass spectrometry peak for each pair of enantiomers eluted from the chiral column is further detected by the detector.

[0074] Furthermore, a preliminary operating mode based on a preliminary operating route (using a standard substance) determines the retention time of the enantiomer under the condition a and the retention times of multiple pairs of enantiomers under the condition b, and the preliminary operating mode can be performed by the first operating route described above.

[0075] 2 and 3, the detection of a target substance having one chiral center (i.e., a pair of enantiomers) will be described as an example.

[0076] In Figure 2, when detecting a mixture containing both non-chiral and chiral components, the six-way valves V1 and V2 form a first operating path, as shown by the bold black line. The corresponding detection system is in a first operating mode. In this mode, after the sample is supplied by the sample injector, liquid-phase pumps 11 and 12 are operated to supply the mobile phase (including mobile phase A and mobile phase B) of the liquid chromatograph system. At the same time, the mass spectrometer (configured to display or respond only to the characteristic mass spectrometric signal of the target chiral compound) is operated. The sample is separated using reversed-phase liquid chromatography 10. The components eluted from 10 enter the trap ring and then the mass spectrometer detector 40. The mobile phase composition can be adjusted to separate the non-chiral components and enantiomers. The eluted target chiral components enter the trap ring 30 and then the mass spectrometer detector 40, generating a signal response from the mass spectrometer detector. That is, when this response occurs, the operator also recognizes that the target component has actually been collected in the trap ring 30, and switches the positions of V1 and V2 to switch to the second operating path.

[0077] As shown in Figure 3, after communicating with the second operating path, the analytical system enters a second operating mode. In this mode, the supercritical fluid chromatograph system is activated, i.e., the supercritical fluid supply device 21 (supplying mobile phase C) and the liquid phase pump 22 (supplying mobile phase D) are activated, so that the supercritical fluid eluates from the trap ring 30 and is supplied to the chiral column group 20 for enantiomer separation. A pair of enantiomers eluted from the chiral column group 20 is separated by the chiral column and exhibits two separated chromatographic peaks with different retention times for the same characteristic ion in the mass spectrometer detector 40. Furthermore, by comparing the peak intensities or peak areas, the relative contents of the two compounds with different spatial conformations in the enantiomers in the detection target can be determined.

[0078] Furthermore, when the target compound contains a compound with multiple chiral centers, the trap 30 may be replaced with a trap having multiple storage compartments when performing detection as shown in Figure 2. Furthermore, the retention times of the different chiral components may be detected in advance using the above-described preliminary operating pathway / mode to determine the elution times of the different chiral components. For a target compound with three chiral centers, four pairs of enantiomers may be present. Detection using the preliminary operating pathway results in four response signals (one for each pair of enantiomer) appearing at different times on the mass spectrometer detector, allowing the retention times of the four pairs of enantiomers to be determined. The first operating mode described above is then performed, retaining each of the four pairs of enantiomers in a different storage compartment of the trap according to their corresponding retention times. The second operating mode described above is then performed, separating each of the four pairs of enantiomers.

[0079] <Third aspect> In a third aspect of the present invention, there is provided a method for evaluating or screening catalysts used to synthesize chiral compounds having at least one chiral center.

[0080] Here, the evaluation method includes synthesizing the chiral compound using each catalyst to be evaluated, detecting each product obtained from each catalyst using the detection method described in the second aspect, and measuring the ratio of the intensities or areas of two characteristic mass spectrometry peaks of one or each pair of target enantiomers to determine the spatial selectivity of the catalyst.

[0081] In some preferred embodiments of the present invention, the detection method of the present invention comprises: Catalysts for the synthesis of chiral compounds with one chiral center, or • Compounds with multiple chiral centers can be synthesized, but are particularly suited to screening catalysts where the target detectant is only one pair of enantiomers.

[0082] Only one mass spectrometer detector can be used, and the relative proportions of the two enantiomers in the synthesized product can be easily determined by switching a multi-way valve (without predetermining the retention times of the achiral and chiral components in the one-dimensional chromatogram), thus significantly improving the efficiency of catalyst screening.

[0083] The present invention also provides a method for synthesizing a drug containing a chiral compound, wherein the synthesis of the chiral compound includes screening of a catalyst using the evaluation method described above.

[0084] The present invention further provides a method for quality control of a traditional Chinese medicine containing chiral components, which realizes quality control of the traditional Chinese medicine by separating the chiral components and determining the content of the chiral components. [Example]

[0085] Hereinafter, embodiments of the present invention will be described in detail with reference to examples. However, those skilled in the art will understand that the following examples are merely for the purpose of illustrating the present invention and are not intended to limit the present invention. In the examples, unless specific conditions are specified, the experiments were carried out under standard conditions or conditions recommended by the manufacturer. Reagents and equipment used without a manufacturer's name are all commercially available standard products.

[0086] device In this example, a multidimensional chromatography-mass spectrometry system (see Figure 1) was constructed using an ultra-high performance liquid chromatograph Nexera LC-40, a supercritical fluid chromatograph Nexera UC, and a mass spectrometer LCMS-8050 manufactured by Shimadzu Corporation, and a method was developed.

[0087] The specific equipment is as follows: First dimension: Two LC-40XR pumps, SIL-30AC automatic sample injector, CTO-20AC column oven (equipped with two high-pressure two-position six-way valves) Second dimension: LC-30ADSF supercritical CO2 fluid pump, LC-30AD entrainer pump, SFC-30A backpressure control unit, CTO-20AC column oven (equipped with two high-pressure 6-position 7-way valves) CBM-20A system controller, LCMS-8050 triple quadrupole mass spectrometer, LabSolutions LCMS Ver.5.99SP2 workstation. Analysis conditions Liquid phase conditions: 1st dimension: Reversed phase separation column: Shim-pack GIST C18 2.0 x 100 mm, 2 μm Column temperature: 40℃ Reverse phase mobile phase: A: 5 mM NH3HCO3, B: AcCN; flow rate: 0.2 mL / min Elution method: Gradient elution, see Table 1

[0088] [Table 1]

[0089] Second dimension: SFC chiral column: Opti-Chiral C1-5 4.6 x 150 mm, 5 μm Column temperature: 40℃ SFC mobile phase: A: SF-CO2, B: 1% 2M NH3-MeOH; flow rate: 2mL / min Elution method: Gradient elution, see Table 2 BPR: 10 MPa, 50°C

[0090] [Table 2]

[0091] sample: The Yuanhu analgesic tablets were taken, the coating removed, 1 mg was accurately weighed, 1 mL of methanol was added, and the tablets were subjected to ultrasonic extraction. After filtering through a membrane filter, the tablets were analyzed by the instrument. Injection volume: 0.5 μL Sampling (100uL) valve switching time: 4.9min Mass spectrometer valve switching time: 7.0 min

[0092] Mass spectrometry conditions: [Table 3]

[0093] [Table 4]

[0094] Results and Discussion Tetrahydropalmatine reference material analysis A mixed solution of L-tetrahydropalmatine (L-THP) and D-tetrahydropalmatine (D-THP) reference materials was analyzed using an LC-SFC multidimensional chromatographic system. Under first-dimension gradient conditions, the L-THP and D-THP mixture eluted from the reversed-phase column in 4.9 minutes. Furthermore, by switching the valve, the L-THP and D-THP mixture was introduced into the second-dimension SFC, where a chiral column screening system was used. Finally, chiral separation was achieved within 5 minutes using an Opti-Chiral C1 chiral column (see Figure 4).

[0095] Yuanhu Pain Relief Tablet Analysis The methanol extract of Yuanhu analgesic tablets was analyzed using an LC-SFC multidimensional chromatographic system, and the MS detector was switched from the first dimension to the second dimension in 7 minutes. As can be seen from Figure 5, the mixture of L-THP and D-THP was separated from the other isomers in the Yuanhu analgesic tablet extract by 7 minutes. Furthermore, the L-THP and D-THP mixture, which showed a peak at 4.9 minutes, was introduced into the second-dimensional SFC chiral separation system using a high-pressure switching valve, and the enantiomers were rapidly separated within 5 minutes.

[0096] conclusion The LC-SFC multidimensional chromatographic system can rapidly and automatically separate complex samples containing chiral compounds into non-chiral and chiral fractions. This system is applicable to the automated screening of chiral catalysts and the rapid analysis of chiral components in traditional Chinese medicines.

[0097] Although the present invention has been described with reference to specific embodiments, those skilled in the art will understand that the present invention is not limited to these.

[0098] Although various embodiments of the present invention have been described above, the above descriptions are illustrative and not exhaustive, and are not limited to the disclosed embodiments. It will be apparent to those skilled in the art that various modifications and variations are possible without departing from the scope and spirit of the described embodiments. The terms used in this specification have been selected to best explain the principles, actual use, or technical improvements in the market of each embodiment, or to enable other ordinary skilled in the art to understand each embodiment disclosed herein. [Industrial Applicability]

[0099] The analytical system and detection method provided by the present invention can be implemented industrially.

Claims

1. 1. A multi-modal analytical system for use in detecting chiral compounds, comprising: The analytical system includes an injection system, a liquid chromatography system as a first-dimension chromatograph, a supercritical chromatography system as a second-dimension chromatograph, a trap system, a connection system, and a detector; the liquid chromatographic system includes a reversed-phase liquid column, the supercritical chromatographic system includes one or more chiral columns, the connection system includes a first multi-port valve and a second multi-port valve, and the detector is a mass spectrometry detector; The first multi-way valve is configured to: i) providing a connection between said liquid chromatographic system and said trapping system; or ii) providing a connection between said trapping system and said supercritical chromatographic system; The second multi-way valve is configured to switch the valve position so that the mass spectrometric detector: i') detecting the eluate from said liquid chromatographic system; or ii') An analytical system for detecting effluents from the supercritical chromatography system.

2. The analytical system of claim 1, wherein the injection system is an automatic injection system and is in communication with the reversed-phase liquid column.

3. 3. The analytical system according to claim 1, wherein the liquid chromatograph system further comprises a liquid chromatography mobile phase supply device, and the reversed-phase liquid column is a silica gel column modified with a hydrophobic group.

4. 4. The analytical system according to claim 1, wherein the trap system is a trap ring or a trap having one or more receiving portions.

5. 5. The analytical system according to claim 1, wherein the supercritical chromatography system further comprises a supercritical chromatography mobile phase supply device and a back pressure control device.

6. 6. The analytical system according to claim 1, wherein the supercritical chromatography system includes two or more chiral columns.

7. 7. The analytical system according to claim 1, wherein the first multi-port valve and the second multi-port valve are both six-port valves.

8. a first working path in which the reversed-phase liquid column communicates with the trapping system via the first multi-way valve, and the trapping system further communicates with the detector via the second multi-way valve; The analytical system according to any one of claims 1 to 7, further comprising a second operating path in which the trapping system is connected to the supercritical chromatography system via the first multi-way valve, and the chiral column further communicates with the detector via the second multi-way valve.

9. 9. The analytical system of claim 8, wherein the connection in the second working path communicates the trap system with a mobile phase supply of a supercritical chromatography system via the first multi-way valve.

10. 10. The analytical system according to claim 8, further comprising an operating path that uses the first operating path as a backup operating path.

11. 11. The analytical system according to claim 1, wherein the first and second multi-port valves of the connection system are switched between positions by a self-regulating system.

12. 12. The analytical system of claim 1, wherein the mass spectrometric detector is configured to display or respond only to the characteristic mass spectrometric signal of the target chiral compound.

13. 1. A method for detecting a chiral compound, comprising: Detection is performed using the analysis system according to any one of claims 1 to 12, The method for detecting said chiral compound is characterized in that it has one or more chiral centers.

14. The detection method according to claim 13, wherein the chiral compound is a low molecular weight compound.

15. a. When the test sample only has a pair of enantiomer compounds or the target detection product in the test sample is only a pair of enantiomers, after the pair of enantiomers are collected in the trap system according to the corresponding retention time, or after the corresponding characteristic mass spectrometry peak detection signal of the compound appears in the detector, switch the first multi-way valve and the second multi-way valve to enter a second operation mode; b. If the test sample contains two or more pairs of enantiomer compounds, collect each pair of enantiomers into the trap system according to the corresponding retention time, and after the collection is completed, switch the first and second multi-way valves to enter the second operation mode; a first operation mode in which the reversed-phase liquid column is connected to the trapping system via the first multi-way valve, and the trapping system is further connected to the detector via the second multi-way valve, and one or more pairs of enantiomers in a test sample are sequentially collected in the trapping system; The detection method described in claim 13 or 14, characterized in that it includes a second operating mode in which the trapping system is connected to the supercritical chromatography system via the first multi-port valve, and the chiral column is further connected to the detector via the second multi-port valve, and the peak intensity or area of ​​a characteristic mass analysis peak for each pair of enantiomers eluted from the chiral column is further detected by the detector.

16. The detection method described in claim 15, characterized in that the retention times of the enantiomers under condition a and the retention times of the pairs of enantiomers under condition b are determined by a preliminary operation mode, in which the reversed-phase liquid column is connected to the trap system via the first multi-way valve, and the trap system is further connected to the detector via the second multi-way valve.

17. The detection method according to any one of claims 13 to 16, characterized in that the detector detects the eluate from the chiral column and determines the ratio of the peak intensities or areas of characteristic mass spectrometry peaks of two isomers of one or more pairs of enantiomers, thereby determining the content ratio of the two isomers.

18. A method for evaluating a catalyst for synthesizing a chiral compound, comprising: An evaluation method characterized by synthesizing the chiral compound using each catalyst to be evaluated, detecting each product obtained for each catalyst using the detection method according to any one of claims 13 to 17, and measuring the peak intensity or area ratio of the characteristic mass spectrometry peaks of a pair of target enantiomers, thereby evaluating the corresponding catalyst.

19. A method for synthesizing a drug comprising a chiral compound, comprising: A drug synthesis method, characterized in that the synthesis of the chiral compound includes screening of a catalyst using the evaluation method described in claim 18.

20. A method for evaluating the quality of a herbal medicine, comprising: A quality evaluation method for evaluating the quality of a herbal medicine by analyzing the chiral components in the herbal medicine using the detection method according to any one of claims 13 to 17 and measuring the relative proportions of the enantiomers.

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