Tool for preparing crystal structure analysis sample, and method for manufacturing the same, tool for crystal structure analysis, and method for manufacturing the same, and method for determining molecular structure of target molecule

Abstract

To provide a tool for preparing a crystal structure analysis sample and a method for manufacturing the same that allow execution of a series of operations in a molecular structure determination method by a crystal sponge method without contact of an experimental instrument or fingers with single crystals, and contribute to full automation of the series of operations.SOLUTION: A tool for preparing a crystal structure analysis sample has a single crystal holder and one or two or more single crystals. The single crystal holder has a shape that can be fixed to a goniometer of a crystal structure analyzer. The single crystal has a plurality of regularly arranged target molecule accommodation spaces. The single crystals are directly fixed to a surface of the single crystal holder.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to an instrument for preparing a sample for crystal structure analysis and a method for producing the same, an instrument for crystal structure analysis and a method for producing the same, and a method for determining the molecular structure of a target molecule. [Background technology]

[0002] Conventionally, single crystal structure analysis methods such as single crystal X-ray diffraction have been known as one of the highly reliable methods for determining molecular structures. However, since single crystal structure analysis requires the use of high-quality single crystals as measurement samples, when performing crystal structure analysis of unknown compounds or compounds that are difficult to crystallize, it can take a lot of time and effort to prepare the measurement samples.

[0003] As a method for solving this problem, a method using a single crystal of a porous polymer compound is known. For example, Patent Document 1 describes a method in which molecules of a compound to be analyzed are regularly arranged within the pores of a single crystal of a polymer complex, and the resulting guest molecule inclusion complex is used as a measurement sample to perform crystal structure analysis (the so-called "method for determining molecular structure by the crystalline sponge method"). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2014 / 038220 Summary of the Invention [Problem to be solved by the invention]

[0005] Figure 1 is a simple diagram showing an example of the procedure for determining the molecular structure of a molecule of a target compound (hereinafter, the "molecule of the target compound" may be referred to as the "target molecule") using the crystalline sponge method. In this example, first, as shown in Figs. 1(a) and (b), a raw material solution of the polymer complex is prepared, and then allowed to stand to precipitate single crystals of the polymer complex. Next, as shown in Figures 1(c) and (d), a portion of the obtained single crystal is sucked up together with the solution using a pipette or the like, and the single crystal is transferred to a container containing the solution in which the target molecule has been dissolved, and the single crystal is immersed in the solution in which the target molecule has been dissolved. Next, as shown in FIG. 1(e), the solution in which the target molecules are dissolved is concentrated while the target molecules are incorporated into the single crystal. Next, as shown in Figure 1(f) and (g), the single crystal incorporating the target molecule is fixed to the tip of a glass rod using adhesive or the like, or is packed into a capillary and fixed, or is fixed to a sample loop, and then the single crystal is placed in a crystal structure analyzer for measurement.

[0006] As described above, conventional methods require procedures in which laboratory equipment directly contacts the single crystal, such as moving the single crystal with a pipette or fixing the single crystal before performing crystal structure analysis, and these procedures pose the risk of deteriorating the single crystal. Furthermore, in order to perform structural analysis more efficiently, it would be desirable to fully automate the series of steps in the molecular structure determination method using the crystalline sponge method; however, single crystals can be very fragile, and it has been difficult to mechanize tasks in which laboratory equipment, etc., as described above, directly contacts the single crystals.

[0007] The present invention solves these problems, and aims to provide a crystal structure analysis sample preparation instrument and its manufacturing method which enable a series of steps in a molecular structure determination method using the crystalline sponge method to be performed without bringing laboratory equipment or fingers into contact with the single crystal, and which contributes to the complete automation of the series of steps; a crystal structure analysis instrument obtained using the crystal structure analysis sample preparation instrument and its manufacturing method; and a method for determining the molecular structure of a target molecule. [Means for solving the problem]

[0008] In order to solve the above problems, the present inventors have extensively investigated a series of steps in a molecular structure determination method using the crystalline sponge method. As a result, they discovered that by using an instrument having a shape that can be fixed to the goniometer of a crystal structure analysis instrument and a single crystal that functions as a crystalline sponge, it is possible to perform a series of steps in a molecular structure determination method using the crystalline sponge method without touching the single crystal with laboratory equipment or fingers, and that it is possible to fully automate the series of steps, thereby completing the present invention.

[0009] Thus, according to the present invention, there are provided instruments for preparing a sample for crystal structure analysis as described below in [1] to [8], a method for manufacturing an instrument for preparing a sample for crystal structure analysis as described in [9], instruments for crystal structure analysis as described in

[10] and

[11] , a method for manufacturing an instrument for crystal structure analysis as described in

[12] , and methods for determining the molecular structure of a target molecule as described in

[13] to

[15] .

[0010] [1] An apparatus for preparing samples for crystal structure analysis having a single crystal holder and one or more single crystals, wherein the single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analysis apparatus directly or via other parts, the single crystal has a plurality of regularly arranged target molecule accommodation spaces, and the single crystal is directly fixed to the surface of the single crystal holder. [2] The apparatus for preparing a sample for crystal structure analysis according to [1], wherein the shape of the entire or a part of the single crystal holder is capillary-shaped, rod-shaped, wire-shaped, mesh-shaped, multi-hole plate-shaped, dish-shaped, plate-shaped or thin-film-shaped. [3] The apparatus for preparing a sample for crystal structure analysis according to [1] or [2], wherein the single crystal holder is made entirely or in part of glass, resin or carbon. [4] The apparatus for preparing a sample for crystal structure analysis according to any one of [1] to [3], wherein the single crystal is directly fixed to the surface of the single crystal holder by having grown on the surface of the single crystal holder. [5] The apparatus for preparing a sample for crystal structure analysis according to any one of [1] to [4], wherein the single crystal is one in which, when the smallest rectangular parallelepiped capable of containing the single crystal is imagined, the length of the longest side is 500 μm or less. [6] The apparatus for preparing a sample for crystal structure analysis according to any one of [1] to [5], wherein the single crystal is a crystal of a polynuclear metal complex. [7] The apparatus for preparing a sample for crystal structure analysis according to any one of [1] to [6], wherein the single crystal holder is a glass capillary, the single crystal is directly fixed to the inner wall surface of the glass capillary, and the single crystal has a length of the longest side of the smallest imaginary rectangular parallelepiped capable of containing the single crystal, which is 100 μm or less. [8] The apparatus for preparing a sample for crystal structure analysis according to [7], wherein the number of single crystals fixed to the inner wall of the glass capillary is 1 to 1000 per glass capillary. [9] A method for manufacturing an apparatus for preparing a sample for crystal structure analysis according to any one of [1] to [8] above, comprising the steps of contacting a single crystal holder with a solution for preparing a single crystal and precipitating a single crystal on a surface of the single crystal holder.

[10] A crystal structure analysis instrument having a single crystal holder and one or more crystal structure analysis samples, wherein the crystal structure analysis samples are formed by regularly accommodating target molecules in the target molecule accommodating space of a single crystal constituting the crystal structure analysis sample preparation instrument described in any one of [1] to [6].

[11] A crystal structure analysis instrument having a single crystal holder and one or more crystal structure analysis samples, wherein the crystal structure analysis samples are formed by regularly accommodating target molecules in the target molecule accommodating space of a single crystal constituting the crystal structure analysis sample preparation instrument described in [7] or [8].

[12] A method for producing a crystal structure analysis instrument according to

[10] or

[11] , comprising a step (Step I) of contacting target molecules with a single crystal constituting the instrument for preparing a sample for crystal structure analysis according to any one of [1] to [8] above, and regularly accommodating the target molecules in the target molecule accommodating space of the single crystal, characterized in that the operation of Step I is performed without a laboratory instrument or a finger coming into contact with either the single crystal constituting the instrument for preparing a sample for crystal structure analysis or the crystal structure analysis sample constituting the instrument for crystal structure analysis.

[13] A method for determining the molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument described in

[10] or

[11] , characterized in that the crystal structure analysis sample constituting the crystal structure analysis instrument is placed in a crystal structure analysis apparatus without contacting the crystal structure analysis sample with laboratory instruments or fingers.

[14] A method for determining the molecular structure of a target molecule by crystal structure analysis, comprising the following steps 1 and 2: [Step 1] A step of irradiating a crystal structure analysis sample constituting the crystal structure analysis instrument according to the above

[11] with synchrotron X-rays and collecting diffraction intensity data. [Step 2] A step of performing the above step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data.

[15] A method for determining the molecular structure of a target molecule according to

[14] , which is carried out using a plurality of crystal structure analysis tools according to

[11] , characterized in that the plurality of crystal structure analysis tools are arranged in parallel to each other and installed in a crystal structure analysis apparatus in a state forming a pseudo-plane. Effect of the Invention

[0011] According to the present invention, there are provided a crystal structure analysis sample preparation instrument and a method for manufacturing the same, which enable a series of steps in a molecular structure determination method using the crystalline sponge method to be performed without bringing laboratory equipment or fingers into contact with the single crystal, and which contribute to the complete automation of the series of steps, a crystal structure analysis instrument obtained using the crystal structure analysis sample preparation instrument and a method for manufacturing the same, and a method for determining the molecular structure of a target molecule. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 is a simplified diagram of a general method for determining molecular structure by the crystalline sponge method. [Diagram 2]FIG. 1 is a schematic diagram showing an example of an instrument for preparing a sample for crystal structure analysis according to the present invention. [Diagram 3] FIG. 1 is a diagram showing a simplified manufacturing process of a crystal structure analysis device having a capillary-shaped single crystal holder. [Figure 4] 1 is a diagram simply illustrating a manufacturing process of a crystal structure analysis tool having a plate-shaped single crystal holder. [Diagram 5] FIG. 1 is a diagram showing a simplified manufacturing process of a dish-shaped crystal structure analysis tool. [Figure 6] 1 is a photograph of the crystals produced in Example 1. [Figure 7] FIG. 1 shows the molecular structure of [(tpt)2(ZnCl2)3·x(n-hexane)]n obtained in Example 4. [Figure 8] FIG. 1 shows the results of the crystal structure analysis carried out in Example 8. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention will be described in detail below, dividing it into the following sections: 1) an instrument for preparing a sample for crystal structure analysis and a method for producing the same, 2) an instrument for crystal structure analysis and a method for producing the same, and 3) a method for determining the molecular structure of a target molecule.

[0014] 1) Crystal structure analysis sample preparation equipment and its manufacturing method The crystal structure analysis sample preparation device of the present invention is a crystal structure analysis sample preparation device having a single crystal holder and one or more single crystals, wherein the single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analysis apparatus directly or via other parts, the single crystal has a plurality of regularly arranged target molecule accommodation spaces, and the single crystal is directly fixed to the surface of the single crystal holder.

[0015] As used herein, the term "target molecule" refers to a molecule of a compound to be analyzed. The term "target molecule-accommodating space" refers to the space inside the single crystal in which the target molecule is accommodated. The shape of the space is not limited as long as it can accommodate the target molecule, and may be elongated, for example, like a "pore." The term "orderly arranged" in "plurality of target molecule-accommodating spaces regularly arranged" means that the structure of the molecules surrounding the target molecule-accommodating spaces can be determined by crystal structure analysis. In this specification, the term "crystal structure analysis sample" refers to a single crystal in which a target molecule is included in the target molecule accommodation space. In addition, in this specification, the term "single crystal" refers, in principle, to a crystal having single crystallinity and not having a target molecule in the target molecule accommodation space (i.e., a precursor of a crystal structure analysis sample). However, in some cases, the term "single crystal" is used to include a crystal having a target molecule in the target molecule accommodation space (i.e., a crystal structure analysis sample).

[0016] The crystal structure analysis sample preparation device of the present invention is used for preparing a crystal structure analysis sample when determining the molecular structure of a target molecule by the crystal sponge method. In the crystal structure analysis sample preparation device of the present invention, the single crystal is fixed to the surface of the single crystal holder. Therefore, in the series of steps in the molecular structure determination method using the crystal sponge method, the single crystal can be moved by holding the single crystal holder without touching the single crystal with laboratory equipment or fingers.

[0017] [Single crystal holder] The single crystal holder constituting the device for preparing a sample for crystal structure analysis of the present invention functions as a member for fixing a single crystal.

[0018] The single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analyzer directly or via another part. Because the single crystal holder has such a shape, it is possible to omit the task of fixing the crystal structure analysis sample (a single crystal incorporating a target molecule) to the tip of a glass rod or inside a capillary, which has traditionally been performed prior to crystal structure analysis.

[0019] The shape of the single crystal holder that can be fixed to the goniometer of the crystal structure analyzer includes, for example, a capillary shape, a rod shape, a plate shape, etc. However, the shape is not limited to these. When the single crystal holder has a capillary or rod-shaped shape that can be fixed to the goniometer of a crystal structure analysis apparatus, the single crystal holder can be efficiently fixed to the goniometer of a crystal structure analysis apparatus by utilizing the goniometer head of a conventional crystal structure analysis apparatus.

[0020] From the viewpoint of reliably fixing a single crystal to the surface of the single crystal holder and from the viewpoint of irradiating a crystal structure analysis sample derived from the single crystal with X-rays or the like to obtain sufficient diffraction intensity data, it is preferable that the shape of all or a part of the single crystal holder is capillary-shaped, rod-shaped, wire-shaped, mesh-shaped, multi-hole plate-shaped, dish-shaped, plate-shaped or thin-film-shaped. That is, when at least the portion of the single crystal holder to which the single crystal is fixed has the above-mentioned shape, the single crystal is fixed securely and sufficient diffraction intensity data can be easily obtained.

[0021] The single crystal holder is preferably made entirely or partially of glass, resin or carbon. In general, in single crystal structure analysis using X-rays or electron beams, a single crystal is fixed on a substrate made of glass, resin or carbon and then measured. Therefore, by making at least a portion of the single crystal holder from glass, resin or carbon, the single crystal fixed to that portion can be suitably used as a precursor of a crystal structure analysis sample.

[0022] The single crystal holder may be subjected to a surface treatment, such as a chemical treatment with an oxidizing agent, an acid treatment, an alkali treatment, a primer treatment, or a silane compound treatment, or a physical treatment such as a corona discharge treatment or a plasma treatment. In a surface-treated single crystal holder, the single crystal tends to be more securely fixed to the surface.

[0023] [Single crystal] The single crystal constituting the device for preparing a sample for crystal structure analysis of the present invention is a precursor of a sample for crystal structure analysis. The single crystal has a plurality of regularly arranged target molecule accommodating spaces. A crystal structure analysis sample is produced by using the single crystal having a plurality of regularly arranged target molecule accommodating spaces.

[0024] The single crystal is directly fixed to the surface of the single crystal holder. In this specification, "the single crystal is directly fixed" means that the single crystal is fixed to the surface of the single crystal holder without using a fixing agent such as an adhesive or a viscous substance. The single crystal fixed directly to the surface of the single crystal holder includes a single crystal that precipitates in the presence of the single crystal holder and grows on the surface of the single crystal holder.

[0025] When a separately manufactured single crystal is fixed to a substrate such as a single crystal holder using an adhesive or the like, there is a risk that the adhesive components will dissolve during the process of incorporating the target molecule into the single crystal, causing the single crystal to peel off, or that the dissolved adhesive components will be contained in the target molecule containing space of the single crystal, thereby affecting the analysis results. Furthermore, the task of fixing the single crystal with an adhesive or the like involves direct contact of laboratory equipment with the single crystal, which may cause deterioration of the single crystal.

[0026] On the other hand, since the single crystal constituting the crystal structure analysis sample preparation device of the present invention is fixed directly to the surface of the single crystal holder, these problems do not occur in the crystal structure analysis sample preparation device of the present invention.

[0027] The size of the single crystal is not particularly limited. When the smallest rectangular parallelepiped capable of accommodating the single crystal is imagined, the length of its long side (longest side) is usually 500 μm or less. However, it is preferable that the single crystal has a size suitable for the crystal structure analysis device to be used. For example, when performing crystal structure analysis using a commercially available X-ray crystal structure analyzer, assuming the smallest rectangular parallelepiped capable of accommodating a single crystal, the length of its long side is usually 10 to 500 μm, preferably 50 to 500 μm, and the length of its short side (shortest side) is usually 10 to 500 μm, preferably 10 to 200 μm.

[0028] When a plurality of crystal structure analysis samples are irradiated with synchrotron X-rays and the molecular structure of the target compound is determined based on the collected diffraction intensity data, when the smallest rectangular parallelepiped capable of accommodating a single crystal is imagined, the length of its long side is usually 0.1 to 100 μm, preferably 1 to 100 μm, and the length of its short side is usually 0.1 to 100 μm, preferably 1 to 10 μm.

[0029] When performing crystal structure analysis by microelectron diffraction, assuming the smallest rectangular parallelepiped capable of accommodating a single crystal, the length of its long side is usually 0.1 to 1000 nm, preferably 1 to 1000 nm, and the length of its short side is usually 0.1 to 100 nm, preferably 0.1 to 100 nm.

[0030] The single crystal is preferably a crystal of a polynuclear metal complex, since molecular design is relatively easy. Polynuclear metal complexes are metal complexes that contain two or more metal ions and ligands. The polynuclear metal complex used in the present invention is, for example, a metal complex having a three-dimensional structure containing two or more metal ions and a ligand having two or more coordinating sites, and having an internal space.

[0031] The metal ion is not particularly limited as long as it can form a polynuclear metal complex. The metal ion is preferably an ion of an element selected from the group consisting of Ti, Fe, Co, Ni, Cu, Zn, Ru, Rh, Pd, Cd, Os, Ir, and Pt, and more preferably an ion of an element of Group 8, 9, or 10 of the periodic table. The valence of the metal ion is not particularly limited, and is usually 1 to 4, preferably 1 to 3, and more preferably 2.

[0032] The ligands include those that form the framework or walls of the space that contains the target molecule (hereinafter, these may be referred to as "ligand (α)"), and those that have other roles (such as adjusting the charge of the polynuclear metal complex or occupying vacant coordination sites of the metal ion (hereinafter, these may be referred to as "ligand (β)").

[0033] The ligand (α) is preferably a multidentate ligand having a π-conjugated system. When the ligand (α) is a multidentate ligand having a π-conjugated system, a stable target molecule storage space is easily formed. The multidentate ligand having a π-conjugated system is preferably a multidentate ligand containing an aromatic group as a central skeleton. A multidentate ligand containing an aromatic group as a central skeleton is relatively rigid and has excellent planarity, so that a stable target molecule storage space is easily formed. An example of the ligand (α) is one represented by the following formula (1).

[0034] [ka]

[0035] In formula (1), A is an m-valent group having aromaticity. X is a divalent organic group or a single bond directly connecting A and Y. Y is a coordinating atom or a monovalent group containing a coordinating atom. m represents an integer of 2 to 6. Multiple Xs may be different from each other, and multiple Ys may be different from each other.

[0036] The group represented by A typically has 6-100, preferably 6-60, and more preferably 6-30, atoms (excluding hydrogen atoms). Examples of the group represented by A include a six-membered aromatic group, a group in which a plurality of six-membered aromatic groups are linked by single bonds, and a group having a porphyrin skeleton.

[0037] Examples of the six-membered aromatic group include groups having an aromatic ring such as a benzene ring, a triazine ring, a pyridine ring, and a pyrazine ring. The six-membered aromatic group may have a substituent other than -(-XY). Examples of the substituent include an alkyl group having 1 to 10 carbon atoms; and a halogen atom such as a fluorine atom, a bromine atom, or a chlorine atom.

[0038] Examples of the group represented by A include, but are not limited to, the following: In addition, "*" represents a bond (position of bonding to X).

[0039] [ka]

[0040] [ka]

[0041] In the above formula, M represents a metal ion. Examples of the metal ion include the same as those exemplified as the metal ions constituting the polynuclear metal complex. Among these, zinc ions are preferred.

[0042] The group represented by X typically has 1-30, preferably 2-20, and more preferably 2-10, atoms (excluding hydrogen atoms). Examples of the group represented by X include, but are not limited to, the divalent group represented by A, hydrocarbon groups such as methylene, ethylene, 1,2-ethenediyl, 1,2-ethynediyl (acetylene), p-phenylene, and m-phenylene, amide groups (-C(=O)-NH-), ester groups (-C(=O)-O-), oxymethylene groups (-O-CH2-), and oxyethylene groups (-O-CH2CH2-). The group represented by X may be a group formed by bonding two or more of these groups. Examples of such groups include, but are not limited to, the following:

[0043] [ka]

[0044] The coordinating atom represented by Y includes an oxygen atom, a sulfur atom, a nitrogen atom, a phosphorus atom, and the like. The monovalent group represented by Y typically has 1-20, preferably 1-15, and more preferably 1-10, atoms (excluding hydrogen atoms). Examples of the monovalent group represented by Y include a pyridyl group, an amino group, a hydroxyl group, a deprotonated amide group, a carboxylate group, a sulfonate group, a phosphonate group, a dithiocarboxylate group, a cyano group, and groups containing these groups as a substituent. Examples of the monovalent group represented by Y include the following.

[0045] [ka]

[0046] Examples of the ligand (α) include, but are not limited to, the following:

[0047] [ka]

[0048] [ka]

[0049] [ka]

[0050] As the ligand (α), a ligand other than that represented by formula (1) can also be used. Examples of such a ligand include the following.

[0051] [ka]

[0052] The ligand (β) is preferably a ligand having a relatively low molecular weight, since a low molecular weight ligand is less likely to have adverse effects such as steric hindrance on the coordination of the ligand (α). The ligand (β) may be a monodentate ligand or a chelating ligand.

[0053] The monodentate ligand used as the ligand (β) is an oxide ion (O 2- ) and other divalent anions; hydroxide ion (OH - ), chloride ion (Cl - ), bromide ion (Br - ), iodide ion (I - ), thiocyanate ion (SCN - ) and other monovalent anions; water, ammonia, electrically neutral coordinating compounds such as monoalkylamines, dialkylamines, and trialkylamines; and the like.

[0054] Examples of chelating ligands used as ligand (β) include bidentate chelating ligands such as ethylenediamine, N,N'-dimethylethylenediamine, N,N,N',N'-tetramethylethylenediamine, 2,2'-bipyridyl, and 1,2-cyclohexanediamine, but are not limited to these.

[0055] The single crystal of the polynuclear metal complex may contain components other than metal ions and ligands. Components other than metal ions and ligands contained in the single crystal of a polynuclear metal complex include, for example, molecules capable of affinity interactions with the ligands, such as π-π interactions, π-σ interactions, etc. Single crystals of polynuclear metal complexes containing such molecules tend to have more stable target molecule accommodation spaces.

[0056] Examples of molecules capable of affinity interactions such as π-π interaction and π-σ interaction with a ligand include compounds represented by the following formulas.

[0057] [ka]

[0058] [ka]

[0059] Examples of the polynuclear metal complexes include those represented by the following formulas (2) to (4).

[0060] [ka]

[0061] In formulas (2) to (4), L 1 represents a ligand represented by the formula (1) in which m is 3 (tridentate ligand), and M 1 represents a divalent metal ion of Groups 8 to 12 of the periodic table, Q represents a monovalent anionic monodentate ligand, "solv" represents a solvent molecule or the like used during synthesis, and T represents a molecule capable of affinity interaction with the ligand, such as π-π interaction or π-σ interaction. a, b, and n are arbitrary numbers.

[0062] Specific examples of the polynuclear metal complexes represented by formulas (2) to (4) include those represented by the following formulas.

[0063] [ka]

[0064] In the formulas (2a) to (4a), "tpt" represents 2,4,6-tris-(4-pyridyl)-1,3,5-triazine, and "solv", "T", a, b, and n have the same meanings as above.

[0065] Details of these polynuclear metal complexes are described in documents such as JP-A-2006-188560, JP-A-2008-214584, and WO2011 / 062260.

[0066] Further, examples of single crystals of polynuclear metal complexes other than those mentioned above include single crystals formed by three-dimensionally regularly assembling host molecules having internal spaces. Examples of host molecules constituting such single crystals include those shown in the diagram below.

[0067] [ka]

[0068] In the above diagram, Pd represents a palladium ion coordinated with a bidentate chelating ligand (eg, ethylenediamine).

[0069] [ka]

[0070] In the above diagram, Pd represents a palladium ion coordinated with a bidentate chelating ligand (eg, ethylenediamine).

[0071] Details of these host molecules are described, for example, in WO 2018 / 159692.

[0072] [Apparatus for preparing samples for crystal structure analysis] 2(a) to (c) are schematic diagrams showing an example of the crystal structure analysis sample preparation device of the present invention. Fig. 2(a) is a schematic diagram of a crystal structure analysis sample preparation device (1) having a single crystal (1b) on the inner wall surface of a capillary-shaped single crystal holder (1a). The single crystal holder (1a) has a portion (1c) that can be fixed to a goniometer of a crystal structure analyzer via, for example, a commercially available goniometer head.

[0073] Fig. 2(b) is a schematic diagram of a crystal structure analysis sample preparation tool (2) having a single crystal (2b) on the surface of a plate-shaped single crystal holder (2a). The single crystal holder (2a) has a portion (2c) that can be fixed to a goniometer of a crystal structure analyzer, for example, via fixing members that sandwich the plate from the front and back sides.

[0074] Fig. 2(c) is a schematic diagram of a crystal structure analysis sample preparation device (3) having a single crystal (3b) on the surface of a single crystal holder (3a) having both a dish-like and a rod-like shape. The single crystal holder (3a) has a portion (3c) that can be fixed to a goniometer of a crystal structure analyzer via, for example, a commercially available goniometer head.

[0075] The crystal structure analysis sample preparation devices (1) to (3) have portions to which a single crystal is not fixed, such as the vicinity of the portions (1c) to (3c) of the single crystal holders (1a) to (3a) that can be fixed to a goniometer of a crystal structure analysis apparatus. Therefore, according to the crystal structure analysis sample preparation device of the present invention, in the series of steps in the molecular structure determination method using the crystalline sponge method, the single crystal can be moved by grasping the single crystal holder without touching the single crystal with laboratory equipment or fingers.

[0076] Furthermore, among the apparatus for preparing a sample for crystal structure analysis of the present invention, the single crystal holder is a glass capillary, the single crystal is directly fixed to the inner wall surface of the glass capillary, and the single crystal has a longest side of 100 μm or less when the smallest rectangular parallelepiped capable of containing the single crystal is imagined (hereinafter, this apparatus for preparing a sample for crystal structure analysis may be abbreviated as "apparatus for preparing a sample for crystal structure analysis (A)"), which is suitably used when preparing a sample for crystal structure analysis to be used when determining the molecular structure of a target molecule by irradiating synchrotron X-rays.

[0077] In other words, when irradiating synchrotron X-rays, multiple fine crystal structure analysis samples are usually prepared, and these crystal structure analysis samples are irradiated with synchrotron X-rays. The molecular structure of the target molecule is determined based on the collected diffraction intensity data. Therefore, the process of incorporating the target molecule into the single crystals also needs to be performed on multiple fine single crystals. With the conventional crystalline sponge method, the experience of the experimenter could have a significant impact on the success of the results.

[0078] On the other hand, when the crystal structure analysis sample preparation tool (A) is used, it is possible to avoid deterioration of the single crystal or the crystal structure analysis sample caused by contact with laboratory tools or fingers. Furthermore, since the number of single crystals can be determined with a certain degree of accuracy before the target molecule is incorporated into the single crystal, crystal structure analysis samples can be produced efficiently even when only a small amount of the target molecule is available.

[0079] In addition, when determining the molecular structure of a target molecule based on diffraction intensity data from multiple crystal structure analysis samples, it is preferable that the crystal structure analysis samples used are homogeneous. In this regard, the crystal structure analysis samples in the same capillary are multiple single crystals crystallized under the same conditions, and the target molecule is included in the single crystals under the same conditions, and are therefore excellent in homogeneity.

[0080] The length of the glass capillary is usually 0.5 to 15 cm, preferably 1 to 8 cm. The diameter (inner diameter) of the part of the glass capillary where the single crystal is fixed is usually 0.1 to 3.0 mm, preferably 0.2 to 1.0 mm. The wall thickness of the glass capillary is usually 5 to 100 μm, preferably 10 to 30 μm.

[0081] The single crystal constituting the crystal structure analysis sample preparation tool (A) has a longest side of 100 μm or less, preferably 0.1 to 100 μm, more preferably 1 to 50 μm, when the smallest rectangular parallelepiped capable of accommodating the single crystal is imagined, and the shortest side is usually 0.1 to 100 μm, preferably 1 to 10 μm.

[0082] The number of single crystals constituting the crystal structure analysis sample preparation tool (A) is usually 1 to 1000, preferably 5 to 500, and more preferably 10 to 100.

[0083] [Method of manufacturing an instrument for preparing a sample for crystal structure analysis] The method for producing an instrument for preparing a sample for crystal structure analysis of the present invention comprises the steps of contacting a single crystal holder with a solution for preparing a single crystal and precipitating a single crystal on the surface of the single crystal holder.

[0084] A single crystal preparation solution is a solution containing components of a single crystal and from which a single crystal is to be precipitated. For example, in the case of a single crystal of a polynuclear metal complex, a solution containing components of the polynuclear metal complex such as metal ions and ligands is used as the single crystal preparation solution. The solution for preparing a single crystal may be a single-phase system, or may be a two-phase system in which two types of solutions are layered on top of each other.

[0085] Regarding the concentration of the solution for preparing a single crystal, for example, in the case of a single crystal of a polynuclear metal complex, the metal ion concentration is usually 1 μM to 1 M, preferably 1 to 100 mM, and the ligand concentration is usually 1 μM to 1 M, preferably 100 μM to 10 mM. The temperature for precipitating the single crystal is usually from -80 to 150°C, and preferably from 10 to 30°C. The standing time for precipitating the single crystals is usually from 1 minute to 1 month, preferably from 1 hour to 1 week.

[0086] In some cases, it is possible to control the size and quality of the resulting single crystal by adjusting the concentration of the solution for preparing the single crystal and the precipitation conditions.

[0087] When the portion of the single crystal holder where the single crystal is fixed is shaped like a capillary, a device for preparing a sample for crystal structure analysis can be obtained by pouring a solution for preparing a single crystal into the capillary. For example, in Fig. 3 (a simplified diagram showing the manufacturing process of a crystal structure analysis tool having a capillary-shaped single crystal holder), a single crystal preparation solution (4) is poured into the capillary-shaped single crystal holder (1a) shown in Fig. 3(a) and allowed to stand in the state shown in Fig. 3(b), resulting in the precipitation of a single crystal (1b) (Fig. 3(c)). Thereafter, the single crystal preparation solution (4) is removed, and cleaning or other treatments are performed as necessary to obtain a crystal structure analysis sample preparation tool (1) (Fig. 3(d)).

[0088] When the shape of the portion of the single crystal holder on which the single crystal is fixed is rod-shaped, wire-shaped, mesh-shaped, plate-shaped, thin-film-shaped, etc., a single crystal preparation solution can be placed in a container and the portion of the single crystal holder on which the single crystal is fixed can be immersed in the solution to obtain an instrument for preparing a sample for crystal structure analysis. For example, in Fig. 4 (a simplified diagram showing the manufacturing process of a crystal structure analysis tool having a plate-shaped single crystal holder), the plate-shaped single crystal holder (2a) in Fig. 4(a) is immersed in a single crystal preparation solution (4) and left to stand in the state shown in Fig. 4(b), whereby a single crystal (2b) precipitates (Fig. 4(c)). Thereafter, the plate-shaped single crystal holder (2a) is taken out and washed as necessary to obtain a crystal structure analysis sample preparation tool (2) (Fig. 4(d)).

[0089] When the shape of the portion of the single crystal holder where the single crystal is fixed is a multi-hole plate, dish, plate, thin film, etc., a solution for single crystal preparation can be poured or applied to the portion of the single crystal holder where the single crystal is fixed, thereby obtaining an instrument for preparing a sample for crystal structure analysis. For example, in Fig. 5 (a simplified diagram showing the manufacturing process of a dish-shaped crystal structure analysis tool), a single crystal holder (3a) having a dish-shaped shape in Fig. 5(a) is filled with a single crystal preparation solution (4) and left to stand in the state of Fig. 5(b), whereby a single crystal (3b) precipitates (Fig. 5(c)). Thereafter, the single crystal preparation solution (4) is removed, and cleaning treatment, etc., is carried out as necessary, to obtain a crystal structure analysis sample preparation tool (3) (Fig. 5(d)).

[0090] The washing treatment can be carried out by bringing a washing solvent into contact with the single crystal or the single crystal holder. The obtained crystal structure analysis sample preparation tool may be subjected to a solvent exchange operation within the single crystal. By carrying out a solvent exchange operation within the single crystal, it may be possible to efficiently accommodate the target molecule. The solvent exchange within the single crystal may be carried out by placing the crystal structure analysis sample preparation tool at rest with the single crystal in contact with the solvent to be exchanged.

[0091] 2) Crystal structure analysis equipment and its manufacturing method The crystal structure analysis instrument of the present invention is an instrument for crystal structure analysis having a single crystal holder and one or more crystal structure analysis samples, wherein the crystal structure analysis samples are formed by a single crystal constituting the instrument for preparing a crystal structure analysis sample, and having target molecules regularly accommodated in its target molecule accommodation space. Thus, the crystal structure analysis instrument of the present invention is similar to the crystal structure analysis sample preparation instrument described above, except that it has a crystal structure analysis sample instead of a single crystal (i.e., a target molecule is present in the target molecule storage space of the single crystal).

[0092] [Crystal structure analysis sample] The crystal structure analysis sample is formed by accommodating target molecules regularly in the target molecule accommodating space of the single crystal constituting the above-mentioned crystal structure analysis sample preparation tool.

[0093] By "the single crystal accommodates the target molecule in an orderly manner" is meant that the single crystal accommodates the target molecule in such a way that the molecular structure of the target molecule can be determined by crystal structure analysis. The single crystal constituting the crystal structure analysis sample preparation tool has a plurality of target molecule accommodation spaces arranged regularly (i.e., has a large number of homogeneous target molecule accommodation spaces). Therefore, as a result of each of the target molecules taken into the single crystal being accommodated in a more stable state, the target molecules as a whole are regularly accommodated.

[0094] The size of the target molecule is not particularly limited as long as the target molecule can enter the target molecule accommodation space of the single crystal. The molecular weight of the target molecule is usually 20 to 3,000, preferably 100 to 2,000.

[0095] The occupancy rate of the target molecule in the crystal structure analysis sample is preferably 20% or more, more preferably 50 to 100%, and even more preferably 80 to 100%. The occupancy rate is a value obtained by crystal structure analysis, and represents the amount of the target molecule actually present in the crystal structure analysis sample, when the amount of the target molecule in an ideal inclusion state is taken as 100%.

[0096] [Instrument for crystal structure analysis] In the conventional crystalline sponge method, the obtained crystal structure analysis sample (single crystal incorporating the target molecule) was fixed to the tip of a glass rod using adhesive or the like, or was packed into a capillary and fixed, or was fixed to a sample loop, and then the crystal structure analysis sample was placed in a crystal structure analyzer for measurement. This may result in laboratory equipment or fingers coming into contact with the crystal structure analysis sample, which may cause the crystal structure analysis sample to deteriorate.

[0097] On the other hand, the crystal structure analysis instrument of the present invention has a crystal structure analysis sample fixed to the surface of a single crystal holder having a shape that can be fixed to a goniometer of a crystal structure analysis apparatus, either directly or via other parts, so that the crystal structure analysis sample can be placed on the crystal structure analysis apparatus together with the crystal structure analysis instrument. In this manner, according to the crystal structure analysis instrument of the present invention, the crystal structure analysis sample can be placed in the crystal structure analysis apparatus without the need for fixing the crystal structure analysis sample to a glass rod or the like.

[0098] Among the crystal structure analysis instruments of the present invention, a crystal structure analysis instrument obtained using the crystal structure analysis sample preparation instrument (A) (hereinafter, this crystal structure analysis instrument may be abbreviated to "crystal structure analysis instrument (B)") is suitably used when determining the molecular structure of a target molecule by irradiating synchrotron X-rays.

[0099] That is, when the crystal structure analysis instrument (B) is used, the number and positions of crystal structure analysis samples can be known in advance, so that synchrotron radiation X-rays can be irradiated more efficiently.

[0100] [Method of manufacturing a crystal structure analysis instrument] The method for manufacturing a crystal structure analysis instrument of the present invention includes a step (Step I) of bringing target molecules into contact with a single crystal constituting the instrument for preparing a sample for crystal structure analysis and regularly storing the target molecules in the target molecule storage space of the single crystal, and is characterized in that the operation of Step I is performed without laboratory instruments or fingers coming into contact with either the single crystal constituting the instrument for preparing a sample for crystal structure analysis or the crystal structure analysis sample constituting the instrument for crystal structure analysis.

[0101] Step I can be carried out, for example, by preparing a solution (or suspension) containing the target molecule and contacting the single crystal with this solution (or suspension), or, if the target molecule is a liquid or gas, by directly contacting the single crystal with the target molecule.

[0102] The solvent of the solution (or suspension) containing the target molecule is appropriately selected from among those that do not dissolve the single crystal and dissolve at least a portion of the target molecule.

[0103] Specific examples of the solvent include aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene, 1,2-dichlorobenzene, and nitrobenzene; aliphatic hydrocarbons such as n-butane, n-pentane, n-hexane, and n-heptane; alicyclic hydrocarbons such as cyclopentane, cyclohexane, and cycloheptane; nitriles such as acetonitrile and benzonitrile; sulfoxides such as dimethyl sulfoxide (DMSO); amides such as N,N-dimethylformamide and n-methylpyrrolidone; ethers such as diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, and 1,4-dioxane; alcohols such as methanol, ethanol, and isopropyl alcohol; ketones such as acetone, methyl ethyl ketone, and cyclohexanone; cellosolves such as ethyl cellosolve; halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride, and 1,2-dichloroethane; nitros such as nitromethane and nitroethane; esters such as methyl acetate, ethyl acetate, ethyl lactate, and ethyl propionate; water; and the like. These solvents can be used alone or in combination of two or more.

[0104] The temperature when performing step I is usually −20 to 150° C., preferably 0 to 50° C. The period of step I is usually 1 second to 10 days, preferably 1 second to 5 days, more preferably 1 minute to 1 day.

[0105] In the conventional crystalline sponge method, when a single crystal was moved to bring it into contact with a target molecule, the solution was sucked up using a pipette or other device, making it impossible to avoid contact between the single crystal and the experimental equipment.

[0106] On the other hand, in the manufacturing method of the crystal structure analysis instrument of the present invention, the operation of step I can be performed without laboratory instruments or fingers coming into contact with either the single crystal constituting the instrument for preparing a sample for crystal structure analysis or the crystal structure analysis sample constituting the instrument for crystal structure analysis.

[0107] When the shape of the part of the single crystal holder to which the single crystal is fixed is rod-shaped, wire-shaped, mesh-shaped, plate-shaped, thin film-shaped, etc., and the single crystal is moved into a container containing a solution containing the target molecule, by grasping the part of the single crystal holder where the single crystal is not present, it is possible to avoid experimental equipment, etc., coming into contact with the single crystal. For example, as shown in Figures 4(d) and (e), in the case of a crystal structure analysis sample preparation tool (2) having a plate-shaped single crystal holder, the edge of the tool can be grasped to move the single crystal into a container containing a solution (5) containing the target molecule. By leaving the single crystal in the state shown in Figure 4(e), the target molecule is taken up into the single crystal (2b), a crystal structure analysis sample (2d) is generated (Figure 4(f)), and a crystal structure analysis tool is obtained (Figure 4(g)).

[0108] Furthermore, when the shape of the portion of the single crystal holder on which the single crystal is fixed is a multi-hole plate, dish, plate, thin film, capillary, etc., the operation of step I can be performed by applying or injecting a solution containing the target molecule, and therefore there is no need to move the single crystal in the first place. For example, as shown in Figures 3(d), (e), and (f), in the case of a crystal structure analysis sample preparation tool (1) having a capillary-shaped single crystal holder, the operation of step I can be performed by pouring a solution (5) containing the target molecule into the capillary and leaving it in that state, thereby obtaining a crystal structure analysis tool (Figure 3(g)). Furthermore, as shown in Figures 5(d), (e), and (f), in the case of a crystal structure analysis sample preparation tool (3) having a dish-shaped single crystal holder on which the single crystal is fixed, the operation of step I can be performed by filling the dish with a solution (5) containing the target molecule and leaving it in that state, thereby obtaining a crystal structure analysis tool (Figure 5(g)).

[0109] 3) Method for determining the molecular structure of a target molecule The method for determining the molecular structure of a target molecule of the present invention is a method for determining the molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument, and is characterized in that the crystal structure analysis sample constituting the crystal structure analysis instrument is placed in a crystal structure analysis apparatus without laboratory instruments or fingers coming into contact with the crystal structure analysis sample.

[0110] As described above, the crystal structure analysis instrument used in the method for determining the molecular structure of a target molecule of the present invention has a crystal structure analysis sample fixed to the surface of a single crystal holder having a shape that allows it to be fixed to a goniometer of a crystal structure analysis apparatus, either directly or via other parts, so that the crystal structure analysis sample can be placed on the crystal structure analysis apparatus without laboratory equipment or fingers coming into contact with the crystal structure analysis sample that constitutes the crystal structure analysis instrument.

[0111] After placing the crystal structure analysis sample in the crystal structure analyzer, the crystal structure analysis sample is irradiated with X-rays or electron beams in accordance with standard methods, and the molecular structure of the target molecule can be determined based on the obtained diffraction intensity data.

[0112] X-rays are electromagnetic waves with wavelengths of approximately 0.05 to 0.25 nm. In an X-ray tube, thermoelectrons emitted from a filament are accelerated in an electric field and collided with an anode (target: a metal such as Cu or Mo), generating two types of X-rays: continuous X-rays and characteristic X-rays. When thermal electrons collide with a metal anode, K-shell electrons are ejected, creating vacant orbitals. When an outer shell electron transitions to this vacant orbit, X-rays with energy equal to the energy difference between the electron orbitals are emitted. The wavelength of the X-rays is unique to the target atoms, so they are called characteristic X-rays, and these characteristic X-rays are used in crystal structure analysis.

[0113] An electron beam is a stream of electrons moving in a certain direction, and is usually obtained by accelerating thermal electrons in an electric field. While X-rays are a flow of particles (photons) that have no electric charge, electron beams are a flow of particles (electrons) that have a negative charge. Therefore, when an electron beam is irradiated onto a sample, it strongly interacts with the atomic nuclei of the atoms that make up the sample and the electrons around them due to the Coulomb force, so that in crystal structure analysis using an electron beam (microelectron diffraction method), sufficient diffraction intensity data can be obtained even for thin and small crystals that are not suitable for X-ray crystal structure analysis. The electron beam used has an acceleration voltage of preferably 10 to 1000 kV, more preferably 100 to 400 kV.

[0114] In the method for determining the molecular structure of a target molecule of the present invention, when the crystal structure analysis instrument used is the crystal structure analysis instrument (B), the molecular structure of the target molecule can usually be determined by a method having the following steps 1 and 2. [Step 1] A step of irradiating a crystal structure analysis sample constituting a crystal structure analysis instrument (B) with synchrotron radiation X-rays and collecting diffraction intensity data. [Step 2] A step of performing the above step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data.

[0115] Step 1 is a step of irradiating a crystal structure analysis sample constituting a crystal structure analysis instrument (B) with synchrotron X-rays and collecting diffraction intensity data. Since the crystal structure analysis sample constituting the crystal structure analysis instrument (B) is very small, when such a crystal structure analysis sample is irradiated with X-rays, it tends to be difficult to obtain sufficient diffraction intensity data. Also, the scattering background due to the materials around the crystal structure analysis sample tends to be relatively high.

[0116] For these reasons, when a general X-ray crystal structure analyzer is used, there is a risk that it may not be possible to determine the molecular structure of the target molecule in the crystal structure analysis sample that constitutes the crystal structure analysis instrument (B); however, by irradiating the crystal structure analysis sample that constitutes the crystal structure analysis instrument (B) with synchrotron X-rays, sufficient diffraction intensity data can be collected and the molecular structure of the target molecule can be determined.

[0117] Crystal structure analysis using synchrotron radiation X-rays is a known technique for determining the molecular structures of biopolymers such as proteins, inorganic compounds, organic compounds, etc. In particular, crystal structure analysis using synchrotron radiation X-rays has the characteristic that sufficient diffraction intensity data can be obtained even if the measurement sample is a microcrystal, and is therefore preferably used when determining the molecular structure of a target molecule in a crystal structure analysis sample constituting the crystal structure analysis instrument (B).

[0118] Synchrotron radiation X-rays are X-rays with high brightness and small beam size, making them suitable for structural analysis of microcrystals. The synchrotron radiation X-rays used have a beam size of 0.1×0.1~100×100μm at the sample position. 2 Preferably, the size is 1×1 to 10×10 μm. 2 is more preferred. The synchrotron radiation X-rays used have a photon flux of 1×10 11 Photons / s or more is preferable, 1×10 12 ~5×10 13 photons / s is preferred.

[0119] Step 2 is a step of performing step 1 on a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data. Step 2 also utilizes known techniques in crystallography using synchrotron X-rays. In other words, in the past, when a sample was irradiated with synchrotron X-rays, the sample would sometimes deteriorate during the irradiation, so molecular structures were determined by irradiating multiple samples with synchrotron X-rays and integrating the data obtained. In recent years, programs used for integrating data have been developed.

[0120] For example, small-wedge data collection is a scheme in which multiple samples are used to collect vibration photographic data at angles of about 5 to 10° per sample. When data processing is performed manually, the data can be selected using BLEND (a program that performs hierarchical clustering based on lattice constants) and then merged.

[0121] In recent years, data collection schemes have also been automated. For example, the SPring-8 beamline BL32XU has been automated as the ZOO system. The ZOO system is an automatic data collection system that combines several component technologies, including KUMA, a technology that collects high-quality data by reducing radiation damage to sample crystals; a high-speed two-dimensional scanning system that uses micro-X-rays to scan and detect crystals that cannot be seen by optical microscope observation; SHIKA, a system that identifies the location of crystals by analyzing the large number of diffraction images output from the scanning system; and KAMO, a technology that almost automatically outputs diffraction data as data to be used for structural analysis.

[0122] When determining the molecular structure of a target molecule in a crystal structure analysis sample that constitutes the crystal structure analysis instrument (B), steps 1 and 2 can be performed more efficiently by using an automatic data collection system such as the ZOO system.

[0123] The method for determining the molecular structure of a target molecule, which comprises steps 1 and 2, can be carried out using a plurality of crystal structure analysis tools (B). It is preferable that the multiple crystal structure analysis tools (B) are arranged in parallel to each other and installed in the crystal structure analysis apparatus in a state where they form a pseudo-plane. A pseudo-plane is formed by multiple crystal structure analysis tools (B), allowing the synchrotron radiation X-rays to be efficiently irradiated onto the crystal structure analysis sample. EXAMPLES

[0124] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to the following examples in any way.

[0125] Example 1: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Manufacturing A suspension containing 20.2 mg of 2,4,6-tris-(4-pyridyl)-1,3,5-triazine (tpt) and 14.4 ml of a mixed solvent of chloroform / methanol = 12:1 was sonicated to obtain a solution with a tpt concentration of 4.48 mM [tpt solution (1a)]. A mixed solvent of chloroform / methanol = 12:1 was prepared as a dilution solvent, and tpt solution (1a) was diluted with this mixed solvent to obtain a solution with a tpt concentration of 2.24 mM [tpt solution (1b)]. A 30 mM ZnCl2 methanol solution [ZnCl2 solution (2a)] and a 50 wt% RbOH aqueous solution were prepared, and 1 ml of the ZnCl2 solution (2a) was mixed with 2.36 μl of the 50 wt% RbOH aqueous solution to obtain a solution with an RbOH concentration of 20 mM and a ZnCl2 concentration of 30 mM [metal ion solution (3a)]. 300 μl of tpt solution (1b) and 33.6 μl of metal ion solution (3a) were mixed and stirred using a vortex. 20 μl of the resulting solution was poured into the opening of a capillary (Hampton Research, Glass Number 50 Capillary HR6-112) using a pipette, and then the solution was allowed to reach the tip of the capillary using a hand-cranked centrifuge. The capillary was left to stand in a slightly tilted state in a lidded test tube for several hours to crystallize the polynuclear metal complex. As a result, the formation of several dozen crystals was confirmed in the capillary. The size (long side length) of the crystals was 100-200 μm. Some of the crystals were overlapping, but some were isolated. FIG. 6 shows a photograph of the capillary after crystal formation (FIG. 6(a)) and an enlarged photograph of the crystal (FIG. 6(b)).

[0126] Example 2: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Manufacturing A solution with a tpt concentration of 1.49 mM [tpt solution (1c)] was prepared, and the polynuclear metal complex was crystallized in the same manner as in Example 1, except that tpt solution (1c) was used instead of tpt solution (1b) and the amount of metal ion solution (3a) used was changed to 22.4 μl. As a result, the formation of several dozen crystals was confirmed in the capillary. The size (long side length) of the crystals was about 100 μm. Some of the crystals were overlapping, but some were isolated.

[0127] Example 3: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Manufacturing A solution with a tpt concentration of 0.896 mM [tpt solution (1d)] was prepared, and the polynuclear metal complex was crystallized in the same manner as in Example 1, except that tpt solution (1d) was used instead of tpt solution (1b) and the amount of metal ion solution (3a) used was changed to 13.4 μl. As a result, the formation of several dozen crystals was confirmed in the capillary. The size (long side length) of the crystals was 20 to 50 μm, and almost no overlapping of the crystals was observed.

[0128] Example 4: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Solvent substitution (solvent: n-hexane) and crystal structure analysis of In the same manner as in Example 3, [(tpt)2(ZnCl2)3·x(CHCl3)] was added to the capillary. nA single crystal of was produced. Next, the tip of the capillary was cut with a capillary cutter, and Kimwipe was brought into contact with the solution in the capillary, and the solution in the capillary was drained by capillary action. After that, the capillary was filled with chloroform using capillary action, and the solution in the capillary was drained several times to remove unreacted ligands and metal ions remaining in the capillary. Next, the capillary was immersed in a test tube filled with n-hexane and left at 20°C for one day to perform solvent replacement in the single crystal. Next, the capillary was cut to about 4-5 cm, both ends were sealed with Araldite resin, and the capillary was fixed to the goniometer head with Araldite resin and measured at BL17A, KEK. The wavelength of the synchrotron radiation X-rays used was 0.9 Å, and the camera length was 70 mm. Sixteen of the crystals formed in the capillary were irradiated with synchrotron radiation X-rays, and the diffraction data were processed by XDS software. To obtain sufficient completeness, 12 data sets with high resolution and lattice constants that matched those of the crystalline sponge were integrated by KAMO software. The initial structure was determined by SHELXT, and the structure was refined by SHELXL software. As a result, the host skeleton was observed without applying restraint or constraint commands. Since it was not possible to clearly observe the electron density that could be assigned to the molecular structure in the voids, it was assumed that many hexane molecules were severely disordered, and the effects of the disordered parts were removed by PLATON / SQUEEZE software. [(tpt)2(ZnCl2)3·x(n-hexane)] n The molecular structure of is shown in Figure 7. Figure 7(a) shows the structure of the asymmetric unit, and Figure 7(b) shows the packing as viewed from the b-axis. The molecular skeleton revealed by this crystal structure analysis was found to be the same as that of known crystalline sponges (crystalline sponges formed by conventional methods and solvent replacement).

[0129] Example 5: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Solvent substitution (solvent: cyclohexane) and crystal structure analysis of Solvent replacement in the single crystal and crystal structure analysis were performed in the same manner as in Example 4, except that cyclohexane was used instead of n-hexane as the replacement solvent, and a mixture of protectant oil (Immersion oil for microscopy Type A, Cargille) and cyclohexane was injected into and discharged from the capillary in order to prevent the crystal from drying out before the crystal structure analysis. As a result, it was found that the molecular skeleton revealed by this crystal structure analysis is the same as the molecular skeleton of known crystalline sponges (crystalline sponges formed by conventional methods and solvent replacement).

[0130] Example 6: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Solvent substitution (solvent: ethyl acetate) and crystal structure analysis of Solvent replacement in the single crystal and crystal structure analysis were performed in the same manner as in Example 4, except that ethyl acetate was used instead of n-hexane as the replacement solvent, and a mixture of protective oil (Immersion oil for microscopy Type A, Cargille) and ethyl acetate was injected into and discharged from the capillary in order to prevent the crystal from drying out before the crystal structure analysis. As a result, it was found that the molecular skeleton revealed by this crystal structure analysis is the same as the molecular skeleton of known crystalline sponges (crystalline sponges formed by conventional methods and solvent replacement).

[0131] Example 7: Single crystal [(tpt)2(ZnCl2)3·x(CHCl3)] n Solvent substitution (solvent: 2-butanone) and crystal structure analysis of Solvent replacement in the single crystal and crystal structure analysis were performed in the same manner as in Example 4, except that 2-butanone was used instead of n-hexane as the replacement solvent, and a mixture of protective oil (Immersion oil for microscopy Type A, Cargille) and 2-butanone was injected into and discharged from the capillary in order to prevent the crystal from drying out before the crystal structure analysis. As a result, it was found that the molecular skeleton revealed by this crystal structure analysis is the same as the molecular skeleton of known crystalline sponges (crystalline sponges formed by conventional methods and solvent replacement).

[0132] [Example 8] Inclusion of guest molecule (carvacrol) into single crystal and crystal structure analysis The crystal structure analysis sample preparation tool obtained by the same method as in Example 4 was immersed in a test tube containing carvacrol and stored for one day for soaking treatment. After that, the length was adjusted to a length that would not bend, and both ends were closed with araldite. This was fixed to the goniometer head with araldite resin and measured at KEK BL17A. The wavelength of the synchrotron radiation X-rays used was 0.9 Å, and the camera length was 70 mm. Of the crystal structure analysis samples in the capillary, 15 were irradiated with synchrotron radiation X-rays, and the diffraction data was processed by XDS software. To obtain sufficient completeness, 11 data sets with high resolution and lattice constants that matched those of the crystalline sponge were integrated by KAMO software. The initial structure was determined using SHELXT software, and the structure was refined using SHELXL software. As a result, 9 molecules of carvacrol were observed. SIMU, DFIX, DANG, and ISOR were used as appropriate for the observed guest molecules. Many Q peaks that could not be assigned to molecular structures were considered to be due to severe disorder of the solvent or carvacrol molecules, and the influence of the disordered parts was removed using PLATON / SQUEEZE software. The results are shown in Figure 8. FIG. 8(a) shows the structure of the asymmetric unit, and FIG. 8(b) is an enlarged view of the circled area in FIG. 8(a). Nine carvacrol molecules and one water molecule were identified in the asymmetric unit. The occupancy rates of the observed carvacrol were 1 molecule (100%), 2 molecules (75%), 2 molecules (50%), 1 molecule (40%), 1 molecule (33%), and 2 molecules (25%).

[0133] [ka] [Explanation of symbols]

[0134] 1, 2, 3: Equipment for preparing samples for crystal structure analysis 1a, 2a, 3a: Single crystal holder 1b, 2b, 3b: Single crystal 1c, 2c, 3c: Sites that can be fixed to the goniometer of a crystal structure analyzer 1d, 2d, 3d: Crystal structure analysis sample 4: Solution for single crystal preparation 5: Solution containing target molecule

Claims

1. A crystal structure analysis sample preparation device having a single crystal holder and one or more single crystals, The single crystal holder has a shape that allows it to be fixed to a goniometer of a crystal structure analyzer directly or via another part, The single crystal has a plurality of regularly arranged target molecule-accommodating spaces, The single crystal is directly fixed to the surface of the single crystal holder.

2. 2. The apparatus for preparing a sample for crystal structure analysis according to claim 1, wherein the shape of the entire or a part of the single crystal holder is capillary-like, rod-like, wire-like, mesh-like, multi-hole plate-like, dish-like, plate-like or thin film-like.

3. 2. The crystal structure analysis sample preparation device according to claim 1, wherein the single crystal holder is entirely or partially made of glass, resin or carbon.

4. 2. The crystal structure analysis sample preparation tool according to claim 1, wherein the single crystal is directly fixed to the surface of the single crystal holder by having grown on the surface of the single crystal holder.

5. 2. The crystal structure analysis sample preparation tool according to claim 1, wherein the single crystal has a longest side of 500 μm or less when the smallest rectangular parallelepiped capable of containing the single crystal is imagined.

6. 2. The apparatus for preparing a sample for crystal structure analysis according to claim 1, wherein the single crystal is a crystal of a polynuclear metal complex.

7. the single crystal holder is a glass capillary; The single crystal is directly fixed to the inner wall surface of the glass capillary; 2. The crystal structure analysis sample preparation tool according to claim 1, wherein the single crystal has a longest side of 100 μm or less when the smallest rectangular parallelepiped capable of containing the single crystal is imagined.

8. 8. The crystal structure analysis sample preparation instrument according to claim 7, wherein the number of single crystals fixed to the inner wall of the glass capillary is 1 to 1000 per glass capillary.

9. A method for producing the crystal structure analysis sample preparation instrument according to claim 1, comprising the steps of: A method for manufacturing an instrument for preparing a sample for crystal structure analysis, comprising the steps of contacting a single crystal holder with a solution for preparing a single crystal and precipitating a single crystal on the surface of the single crystal holder.

10. A crystal structure analysis apparatus having a single crystal holder and one or more crystal structure analysis samples, 2. A crystal structure analysis instrument, comprising: a single crystal constituting the crystal structure analysis sample preparation instrument according to claim 1, the single crystal having target molecules regularly accommodated in its target molecule accommodation space.

11. A crystal structure analysis apparatus having a single crystal holder and one or more crystal structure analysis samples, The crystal structure analysis sample is a single crystal constituting the crystal structure analysis sample preparation instrument described in claim 7, which has target molecules regularly accommodated in its target molecule accommodation space.

12. 11. A method for producing a crystal structure analysis instrument according to claim 10, comprising the steps of: bringing target molecules into contact with the single crystal constituting the instrument for preparing a sample for crystal structure analysis according to claim 1; and regularly accommodating the target molecules in the target molecule accommodating space of the single crystal (Step I), A method for manufacturing a crystal structure analysis instrument, characterized in that the operation of step I is performed without laboratory equipment or fingers coming into contact with either the single crystal constituting the instrument for preparing a sample for crystal structure analysis or the crystal structure analysis sample constituting the instrument for crystal structure analysis.

13. A method for determining a molecular structure of a target molecule by crystal structure analysis using the crystal structure analysis instrument according to claim 10, comprising: A method for determining the molecular structure of a target molecule, comprising placing a crystal structure analysis sample constituting a crystal structure analysis tool in a crystal structure analysis apparatus without touching the crystal structure analysis sample with laboratory equipment or fingers.

14. A method for determining the molecular structure of a target molecule by crystal structure analysis, comprising the following steps 1 and 2: Step 1 A step of irradiating a crystal structure analysis sample constituting the crystal structure analysis instrument according to claim 11 with synchrotron X-rays and collecting diffraction intensity data. Step 2 A step of performing the step 1 for a plurality of crystal structure analysis samples and determining the molecular structure of the target molecule based on the collected diffraction intensity data.

15. A method for determining the molecular structure of a target molecule according to claim 14, which is carried out using a plurality of crystal structure analysis instruments according to claim 11, A method for determining the molecular structure of a target molecule, comprising the steps of: placing a plurality of crystal structure analysis tools in a crystal structure analysis apparatus in a state in which the tools are arranged in parallel to each other and form a pseudo-plane;

Images