High performance liquid chromatography separation column for nanoparticle-metal organic framework chiral compound particles and resolution raceme

Abstract

The invention discloses a nanoparticle-metal organic framework chiral compound particle and a high performance liquid chromatography separation column for splitting a chiral compound. In the nanoparticle-metal organic framework chiral compound, chiral ligand nanoparticles are used as a chiral source, and a metal organic framework provides a nano confinement carrier and protection. The chiral ligand nanoparticles and a metal organic framework precursor solution are mixed and reacted to obtain nanoparticle-metal organic framework chiral compound particles with uniformly distributed chiral hot spots, and the nanoparticle-metal organic framework chiral compound particles are coated on the surface of silica gel as a stationary phase to obtain the high performance liquid chromatography separation column for splitting chiral compounds. The high performance liquid chromatographic column provided by the invention shows excellent chiral resolution performance on various different types of chiral compounds or chiral drugs, is wide in application range and high in stability, and shows obvious separation advantages compared with a commercially available Chiralcel AD-H chromatographic column.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of high performance liquid chromatography chiral separation, and particularly relates to a coated chiral nanoparticle metal organic framework high performance liquid chromatography separation column capable of separating a plurality of different types of racemic compounds. BACKGROUND

[0002] Efficient acquisition of a single stereoisomer in a racemic compound has important research significance in the fields of drug treatment and analysis, agriculture and chemical industry. Different stereoisomers of a racemic compound can exhibit different pharmacological or physiological activities in the action on active organisms, which promotes them to exhibit a "chiral preference" phenomenon in the action on active organisms, wherein the "superior isomer" often exhibits a "positive" activity on the active organisms, i.e., can exhibit a drug treatment activity on the active organisms, but the "inferior isomer" opposite to the "superior isomer" often cannot exhibit a positive drug treatment effect on the active organisms, and can exhibit an adverse effect opposite to the treatment of the "superior isomer" on the active organisms, or can even exhibit a serious toxic side effect on the active organisms. For example, in the thalidomide incident that shocked the world in the middle of the last century, the R-(+) configuration of the chiral drug thalidomide has a sedative and anti-emetic effect on pregnant women, effectively alleviating the effects of pregnancy, but the S-(-) configuration of thalidomide has a serious teratogenic effect, which leads to the birth of about 12,000 infants with serious developmental deformities worldwide. This incident makes people realize the importance of exploring the treatment ability of different configurations of chiral drugs on active organisms, and thus it has important research significance in the fields of effective separation of chiral compounds or chiral drugs to obtain a single configuration beneficial to active organisms and avoid harmful configurations, and treatment of active organisms.

[0003] At present, although the existing high performance liquid chromatography commercial columns all have good chiral separation performance, they still cannot separate some racemic compounds. Moreover, the stationary phase materials in these commercial columns also have problems such as complicated and complex synthesis process, high price and cost, etc. Therefore, it is of great significance to develop a high performance liquid chromatography chiral stationary phase which has a simple and easy-to-obtain synthesis process, low preparation cost, excellent chiral recognition and separation performance, and can separate racemic compounds that cannot be separated or cannot be well separated by the existing commercial columns. In recent years, a variety of chiral materials with chiral recognition and separation potential have been developed for use as chromatographic chiral stationary phases. Among them, the metal organic framework material (MOFs) formed by the coordination between the metal center ion or ion cluster and the organic ligand has excellent structure adjustability, and has the advantages of simple and easy-to-obtain synthesis, good stability, adjustable cavity structure and performance, etc., so that it has the ability to coat specific chiral nanoparticle materials, thereby having potential application value in the field of high performance liquid chromatography chiral stationary phase. SUMMARY

[0004] The present application aims at the deficiencies of the prior art, and provides a kind of coated chiral nanoparticle metal organic framework high performance liquid chromatography separation column capable of resolving multiple different types of racemic compounds.The chromatographic column can resolve multiple different types of racemic compounds, including chiral compounds or chiral drugs, and compared with commercially available Chiralcel AD-H chiral high performance liquid chromatography product column, it shows better chiral resolution performance and has a prospect of popularization and application.

[0005] The present application is realized by the following technical solutions: metal organic framework (MOFs) material coated with chiral nanoparticles is effectively prepared by organic reaction, is uniformly distributed on the surface of silica gel by "net wrapping method" to prepare high performance liquid chromatography chiral stationary phase, is prepared into high performance liquid chromatography chiral column by high pressure homogenization method and 30MPa nitrogen pressure, and is applied to realize resolution of multiple different types of racemic compounds.

[0006] In order to achieve the above object, the present application adopts the following technical solutions:

[0007] Preparation of chiral nanoparticles: based on the reaction of chiral ligand and metal ion, chiral nanoparticle (Chiral-NCs) based on chiral ligand is prepared in high-purity water as a reaction solvent under the condition of 50-90 DEG C reaction;

[0008] Preparation of Chiral-NCs@MOFs composite coated with Chiral-NCs: based on the reaction of organic ligand and metal center in the synthesis of MOFs with acid as catalyst, the prepared Chiral-NCs is added to participate in the reaction, and Chiral-NCs@MOFs is prepared in the reaction system;

[0009] Preparation of chiral Chiral-NCs@MOFs stationary phase: in order to make the prepared chiral metal organic framework material more uniformly crosslinked on the surface of silica gel to form a uniform chiral stationary phase, the silica gel and the synthesized chiral metal organic framework material are immersed in a n-hexane solution containing trimesoyl chloride, and after treatment, piperazine aqueous solution is slowly added dropwise.Polymerization and drying obtain Chiral-NCs@MOFs stationary phase;

[0010] Preparation of Chiral-NCs@MOFs chromatographic column: the Chiral-NCs@MOFs stationary phase prepared in step

[0009] is taken, and high pressure homogenization method is adopted to press into a clean high performance liquid chromatography empty column under the pressure of 30Mpa, so that the required Chiral-NCs@MOFs chromatographic column capable of resolving multiple different types of racemic compounds is prepared;

[0011] The chiral Chiral-NCs@MOFs chromatographic column prepared in the application is connected to a high performance liquid chromatograph, and the separation of different types of racemic compounds is realized by optimizing the experimental conditions.

[0012] Compared with the prior art, the application has the following advantages:

[0013] (1) The chiral Chiral-NCs@MOFs chromatographic column has strong chiral separation ability and can separate various types of racemic compounds including chiral compounds or chiral drugs.

[0014] (2) Compared with the commercial Chiralcel AD-H chiral column, the chiral Chiral-NCs@MOFs chromatographic column can separate racemic compounds that cannot be separated by the Chiralcel AD-H chiral column, and has obvious chiral recognition and separation advantages.

[0015] (3) The chiral Chiral-NCs@MOFs separation column has the characteristics of good separation reproducibility, high stability and long service life, and the fixed phase material used is simple to synthesize and has low price, which is conducive to popularization and application. BRIEF DESCRIPTION OF DRAWINGS

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiment or prior art description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments described in the present application, and those skilled in the art can obtain other drawings according to these drawings without creative labor.

[0017] Figure 1 TEM analysis diagram of one of the Chiral-NCs@MOFs complexes used in the application;

[0018] Figure 2 and Table 1 are the separation chromatograms and separation data of part of the racemates using the chromatographic column of the application;

[0019] Figure 3 and Table 2 is a comparison diagram of the separation effect of part of the racemic compounds using the chromatographic column of the application and the commercial Chiralcel AD-H high performance liquid chromatographic column (250mmx4.6mm i.d., Daicel Chiral Technologies, Shanghai). DETAILED DESCRIPTION

[0020] The application will be further described in detail below in conjunction with the drawings and examples, but the drawings and examples are not intended to limit the technical solutions of the application, and all modifications, equivalent transformations and improvements made based on the teaching of the application shall fall within the protection scope of the application.

[0021] Example 1

[0022] Preparation of chiral nanoparticles Chiral-NCs based on chiral ligand: a certain mass of chiral ligand is taken into a 50 mL high-temperature-resistant glass tube, 20 mL of deionized water is added, then metal ions are added, and the reaction is carried out at 50-90°C for 10-24 hours to obtain chiral nanoparticles Chiral-NCs based on chiral ligand;

[0023] Preparation of chiral metal-organic framework composite (Chiral-NCs@MOFs) material: 0.78 mol of organic ligand and 0.51 mol of metal center are respectively taken into a 50 mL high-temperature-resistant glass tube, an acid is used as a catalyst, 5 mL of Chiral-NCs prepared in step

[0023] is added, and the reaction is carried out at 70-100°C for 10-24 hours, and after cooling to room temperature, the product is washed and centrifuged with anhydrous ethanol and deionized water (3 times) to obtain Chiral-NCs@MOFs;

[0024] Preparation of stationary phase: in order to make the prepared chiral metal-organic framework material more uniformly crosslinked on the surface of silica gel balls to form a uniform chiral stationary phase, 3.0 g of silica gel and 0.9 g of the synthesized chiral metal-organic framework composite Chiral-NCs@MOFs are immersed in a n-hexane solution containing trimesoyl chloride (50 m L, 0.05 M), stirred at room temperature for 30 min, and the excess n-hexane solution is removed, then a piperazine (50 m L, 0.5 M) aqueous solution is slowly added dropwise to the silica gel. After polymerization for 30 min, the aqueous solution is poured out, and the solid product is vacuum dried to obtain a chiral Chiral-NCs@MOFs stationary phase;

[0025] The chiral stationary phase material (1.2 g) obtained in step

[0025] is dispersed in a mixture of n-hexane / isopropyl alcohol (9:1, v / v) for 5 minutes. Then the suspension is packed into an empty high-performance liquid chromatography column (150 mm long×2.1 mm i.d.) at 30 MPa by using a high-pressure homogenization method, and n-hexane / isopropyl alcohol (9:1, v / v) is used as a displacement liquid, thereby obtaining a chiral Chiral-NCs@MOFs chromatographic column capable of separating a plurality of different racemic compounds.

[0026] Example 2

[0027] The chiral Chiral-NCs@MOFs separation column obtained in Example 1 is used to separate part of racemic compounds, and the separation chromatograms are as shown inFigure 2 .

[0028] Figure 2 The order of separation of the racemic mixtures and the chromatographic conditions were as follows: (a) benzoin, (b) chlorpheniramine maleate, (c) 1-(9-anthrayl)-2,2,2-trifluoroethanol, (d) DL-phenyllactic acid; the mobile phase was CH3(CH2)4CH3 / (CH3)2CHOH (9:1 / v:v); the flow rate of the mobile phase was 0.1 mL / min. Figure 2 It can be seen that the chromatographic column of this invention enables baseline separation of many different types of racemic compounds, demonstrating excellent chiral recognition and separation capabilities. Some analytical results are shown in Table 1:

[0029] Table 1: Resolution results of some racemic compounds by the chromatographic column of the present invention

[0030]

[0031] Chromatographic parameters: CH3(CH2)4CH3 / (CH3)2CHOH (9:1 / v:v) was used as the mobile phase, the flow rate was 0.1 mL / min, the detector was an ultraviolet detector, the detection wavelength was 210~254nm, and the column size was 15cm×2.1mm id.

[0032] Example 3

[0033] Using the chiral Chiral-NCs@MOFs separation column obtained in Example 1 and the existing commercial Chiralcel AD-H high-performance liquid chromatography column, separation experiments were performed on racemic chlorpheniramine maleate under suitable chromatographic conditions. The chromatograms comparing the separation performance of the column of the present invention with the widely used commercial Chiralcel AD-H high-performance liquid chromatography column are attached. Figure 3 Compare the chromatographic data in Table 2.

[0034] Table 2: Comparative analysis data of the chromatographic column of this invention and the commercial chiral column Chiralcel AD-H.

[0035]

[0036] Chromatographic parameters: The column used in this invention is 150 mm × 2.1 mm id, while the commercial Chiralcel AD-H column is 250 mm × 4.6 mm id. The mobile phase is CH3(CH2)4CH3 / (CH3)2CHOH (9:1 / v:v). The flow rate is 0.1 mL / min for the column used in this invention and 0.5 mL / min for the commercial Chiralcel AD-H column. The detector is an ultraviolet detector. The detection wavelength is 210–254 nm.

[0037] From Table 2 and Figure 3 It is known that chlorpheniramine maleate was not separated on the existing commercial Chiralcel AD-H high-performance liquid chromatography column, while the column of the present invention can separate these two chiral drugs very well, demonstrating that the column of the present invention has a significant advantage in chiral separation compared with the commercial Chiralcel AD-H high-performance liquid chromatography column.

[0038] The foregoing has shown and described the basic principles and main features of the present invention, as well as its advantages. It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered illustrative and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0039] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A high-performance liquid chromatography (HPLC) column for separating racemic chiral compounds, characterized in that: The separation column was filled with chiral nanoparticle-metal-organic framework (MOFs) composite particles (Chiral NCs@MOFs) as the stationary phase.

2. The high-performance liquid chromatography column capable of separating multiple types of racemic chiral compounds according to claim 1, specifically prepared as follows: (1) Preparation of ChiralNCs@MOFs: ① Chiral ligand nanoparticles (Chiral NCs): Thiol-based chiral ligands react with metal ions in deionized water to form Au-S bonded complexes. The reaction continues at 50–90 °C for 10–24 hours to generate Chiral NCs, characterized by: Size: 1–5 nm. The thiol chiral ligand is selected from at least one of L- / D-cysteine ​​and glutathione; the metal ion is selected from at least one of Au, Cu, and Ag. ②Chiral NCs@MOFs: Chiral NCs from ① are mixed with MOF precursors including organic ligands and metal ion solutions, and reacted at 70-100℃ for 10-24 hours to generate Chiral NCs@MOFs. The characteristic is that Chiral NCs form uniformly distributed chiral hot spots in the complex particles. The organic ligand is selected from at least one of terephthalic acid and 4,4'-biphenyl phthalic acid; the metal ion is selected from Zr. 4 + . (2) Preparation of the stationary phase for separation column: Chiral NCs@MOFs from (1) are mixed with silica gel and immersed in a pyromellitic chloride-n-hexane and piperazine-aqueous solution. After a polymerization and crosslinking reaction for 0.5-1 hours, the mixture is separated and dried to obtain a chiral stationary phase coated with Chiral NCs@MOFs@SiO2. The characteristic of this phase is that Chiral NCs@MOFs are uniformly coated on the surface of SiO2. (3) Preparation of high performance liquid chromatography separation column: The chiral silica gel stationary phase coated with Chiral NCs@MOFs in (2) is injected into an empty column by high pressure homogenization at 30 MPa to obtain the ChiralNCs@MOFs@SiO2 chromatographic column that can separate a variety of chiral compounds or chiral drugs.

3. A chiral separation column of chiral NCs@MOFs as described in claim 1, characterized in that: Used for chiral separation analysis in high-performance liquid chromatography, it can efficiently separate different types of racemic chiral compounds, including various chiral drugs and chiral small molecule intermediates.

4. The present invention will be further described in detail with reference to the accompanying drawings and embodiments. However, the drawings and embodiments are not intended to limit the technical solutions of the present invention. All modifications, equivalent transformations, and improvements made based on the teachings of the present invention shall fall within the protection scope of the present invention.

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