CN111398181A - Method for measuring ee value of chiral molecule - Google Patents
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
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Abstract
The invention discloses a method for measuring ee value of chiral molecules, which comprises the steps of preparing a main body solution containing preset main body molecules; adding a guest solution of chiral molecules to be measured with saturated concentration into the host solution to obtain a solution to be detected with saturated CD signals; and measuring a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate the ee value of the chiral molecule to be detected. Compared with the traditional chromatography, the method for measuring the ee value by using the CD spectrum has the advantages that less detection solvent is required, the detection cost is reduced, the ee value of the chiral molecule can be detected more quickly and conveniently, and the method is suitable for screening of high-flux asymmetric catalytic reaction conditions and real-time monitoring of catalytic reaction.
Description
Technical Field
The invention relates to the technical field of chemical property detection, in particular to a method for measuring an ee value of a chiral molecule.
Background
Chirality is a phenomenon commonly occurring in nature (the term chirality means that an object cannot coincide with its mirror image, e.g. our hands, the left hand does not coincide with the right hand, which is a mirror image of each other). In addition to naturally occurring, artificially synthesized compounds such as drugs, pesticides, food additives, etc. also have chirality. Enantiomers of chiral compounds have similar chemical and physical properties, but exhibit distinct pharmacological and physiological activities. Because of the important significance of chiral compounds in the aspects of life science, medicine development and the like, the synthesis of chiral compounds through asymmetric catalytic reaction has become a hot point of chemical research.
The enantiomeric excess value (ee value for short) is an evaluation standard of the effect of the asymmetric catalytic reaction, so that the detection of the ee value is important in the screening of asymmetric catalytic conditions, namely, the control of the ee value directly influences whether the chiral compound can be successfully synthesized through the asymmetric catalytic reaction. The chiral molecular ee value is traditionally detected mainly by adopting a chromatography, although the method has high accuracy, the detection process is long and consumes a solvent, the time consumption is long, and the cost is high, so that the method is not suitable for high-flux asymmetric catalytic condition screening and real-time monitoring of catalytic reaction.
Disclosure of Invention
The invention mainly aims to provide a chiral molecule ee value measuring method, and aims to solve the technical problems that the traditional method for detecting the chiral molecule ee value is long in time consumption and high in cost, and is not suitable for high-flux asymmetric catalytic condition screening and real-time monitoring.
In order to achieve the above object, the present invention provides a method for measuring ee value of chiral molecule, comprising the following steps:
preparing a main body solution containing preset main body molecules;
adding an object solution of chiral molecules to be measured with saturated concentration into the host solution to obtain a solution to be detected with saturated Circular Dichroism (CD, Circular Dichroism) spectrum signals;
and measuring a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate the ee value of the chiral molecule to be detected.
Further, after the step of preparing the host solution containing the predetermined host molecule, the method further comprises:
adding a single chiral object solution into the host solution to obtain a mixed solution saturated with CD signals;
and drawing the preset standard curve equation based on the mixed solution and the standard object solution with the known ee value.
Further, the step of drawing the preset standard curve equation based on the mixed solution and the standard guest solution with the known ee value includes:
detecting the saturated concentration of the CD signal of the mixed solution, and adding a standard guest solution with a known ee value under the saturated concentration into the host solution;
reading a second CD signal intensity value of the host solution added with the standard guest solution at the preset position of the CD spectrum;
and drawing a standard curve by taking the ee value of the standard object solution as a horizontal coordinate and the second CD signal intensity value as a vertical coordinate so as to obtain the preset standard curve equation.
Further, the preparation of the host solution, the chiral guest solution and the standard guest solution is performed in an aqueous solution.
Further, the chiral guest solution of single chirality includes a chiral guest solution of R chirality and a chiral guest solution of S chirality.
Further, the preset host molecule is an amide molecule tube.
Further, after the step of determining a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate an ee value of the chiral molecule to be measured, the method further includes:
recovering the amide molecule tube from the solution to be detected for preparing a new main solution.
Further, the step of recovering the amide molecule tube from the solution to be detected comprises:
adjusting the pH value of the solution to be detected to precipitate to obtain the amide molecular tube;
and carrying out ultrasonic cleaning and drying treatment on the preset main molecules obtained by precipitation so as to complete the recovery of the amide molecule tube.
Further, the concentration of the host solution is between 0.01mM and 0.1 mM.
Further, the preset position is between 250nm and 260nm of the CD spectrum.
The method for measuring the ee value of the chiral molecule comprises the steps of preparing a main body solution containing preset main body molecules; adding a guest solution of chiral molecules to be measured with saturated concentration into the host solution to obtain a solution to be detected with saturated circular dichroism spectrum CD signals; and measuring a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate the ee value of the chiral molecule to be detected.
The invention relates to a method for determining chiral molecule configuration based on CD spectrum, which comprises the steps of adding an object solution of chiral molecules to be measured with saturated concentration into a pre-prepared main body solution containing preset main body molecules to form a solution to be detected with saturated CD signals, then determining a first CD signal intensity value of the solution to be detected at a preset position of the CD spectrum, and substituting the first CD signal intensity value into a pre-drawn and calculated preset standard curve equation, thereby calculating the ee value of the chiral molecules to be measured.
Compared with the traditional mode of measuring by adopting a chromatography, the method for measuring the ee value of the chiral molecule by using the CD spectrum has the advantages that less detection solvent is required, the detection cost is reduced, the ee value of the chiral molecule can be detected more quickly and simply, and the method is suitable for high-flux asymmetric catalytic condition screening and real-time monitoring of catalytic reaction.
Drawings
FIG. 1 is a schematic flow chart of an embodiment of a method for measuring ee value of chiral molecule according to the present invention;
FIG. 2 is a schematic flow chart of another embodiment of the method for measuring ee value of chiral molecule of the present invention;
FIG. 3 is a schematic flow chart of a method for measuring ee value of chiral molecule according to another embodiment of the present invention;
FIG. 4(1a) is a schematic diagram of a predetermined host molecule in an embodiment of the method for measuring ee of a chiral molecule according to the present invention;
FIG. 4(1b) is a schematic diagram showing stereoisomers of a predetermined host molecule in an example of the method for measuring ee of a chiral molecule according to the present invention;
FIG. 5 is a schematic diagram of a representative structure of a chiral molecule capable of detecting a portion of the chiral molecule in an embodiment of a method for measuring an ee value of the chiral molecule according to the present invention.
The implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it should be understood that the specific embodiments described herein are only used for explaining the present invention, and are only a part of the embodiments of the present invention, rather than all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention.
The invention provides a method for measuring an ee value of a chiral molecule.
Referring to fig. 1, fig. 1 is a schematic flow chart of a method for measuring ee of chiral molecules according to a first embodiment of the present invention.
While a logical order is shown in the flow chart, in some cases, the steps shown or described may be performed in an order different than that shown or described herein.
The method for measuring the ee value of the chiral molecule comprises the following steps:
step S100, preparing a host solution containing preset host molecules.
In this embodiment, the preparation of the main solution is performed in an aqueous solution to reduce the detection cost of the ee value of the chiral molecule; in addition, in this embodiment, the preset host molecule is an amide molecule tube having a structure shown in fig. 4(1a), or a stereoisomer of the amide molecule tube shown in fig. 4(1b), and the amide molecule tube having a structure shown in fig. 4(1a) may be selected to prepare a host solution, or the amide molecule tube having a stereoisomer shown in fig. 4(1b) may be selected to prepare a host solution, as required, for different chiral molecules.
In an aqueous solution, a host solution is prepared having a predetermined concentration of a predetermined host molecule.
In this embodiment, the predetermined concentration is a concentration of the prepared host solution, and the concentration is between 0.01mM (millimolar) and 0.1mM, such as 0.01mM, 0.02mM, 0.03mM, 0.04mM, 0.05mM, 0.06mM, 0.07mM, 0.08mM, 0.09mM, or 0.1 mM.
Specifically, for example, 0.56mg of an amide molecule tube having a structure shown in FIG. 4(1a) or FIG. 4(1b) was taken and introduced into a 10m L volumetric flask, and dissolved with double ultrapure water to prepare a bulk solution having a concentration of 0.05mM for the subsequent measurement of ee value of chiral molecule.
In this embodiment, an amide molecular tube having a structure as shown in fig. 4(1a) or a stereoisomer as shown in fig. 4(1b) is selected to prepare a host solution, and the organic small molecules in the solution can be identified based on the hydrophobic effect and the hydrogen bond of the amide molecular tube, so that the host solution prepared by the amide molecular tube can be used for detecting ee values of molecules such as chiral epoxy, chiral alcohol, chiral amine, chiral imine, chiral ester, and chiral ether (fig. 5 shows a representative structure of each chiral molecule from left to right in sequence), and the universality of optical measurement of the ee value of the chiral molecule is improved.
Further, referring to fig. 2, in another embodiment of the method for measuring ee of chiral molecule according to the present invention, after step S100, the method for measuring ee of chiral molecule further comprises:
and S400, adding a single chiral object solution into the host solution to obtain a mixed solution saturated with CD signals.
It should be noted that, in this embodiment, the chiral guest solution of single chirality includes a chiral guest solution of R chirality and a chiral guest solution of S chirality, where the R chirality and the S chirality respectively correspond to two standard configurations of chiral molecules (i.e. for example, the right hand and the left hand of a human hand, and the R corresponds to the right, and the S corresponds to the left).
And gradually adding a pre-prepared chiral object solution with R chirality or S chirality into the prepared host solution with a preset concentration and containing preset host molecules to obtain a mixed solution with saturated CD signals.
Specifically, for example, a prepared chiral guest solution of R chirality is sequentially added to a prepared host solution with a concentration of 0.05mM, and the same prepared chiral guest solution of R chirality is previously added to the aqueous solution to form a mixed solution of the host solution and the chiral guest solution of R chirality, and during the process of sequentially adding the chiral guest solution of R chirality, CD detection is continuously performed on the formed mixed solution until a strongest CD signal is detected (i.e., a CD signal is saturated and is not enhanced with the addition of the chiral guest solution of R chirality), and the continuous addition of the chiral guest solution of R chirality is stopped, so as to obtain a mixed solution of the host solution saturated with the CD signal and the chiral guest solution of R chirality.
In this embodiment, a mixed solution saturated with CD signals is prepared by adding a chiral guest solution of a single chirality (R chirality or S chirality) to a host solution, so that the absolute chirality (R chirality or S chirality) and the ee value of the target substance of the chiral molecule to be measured can be detected simultaneously by the signal intensity and the negativity of the signal of the circular dichroism spectrum.
And step S500, drawing the preset standard curve equation based on the mixed solution and the standard object solution with the known ee value.
Adding a pre-prepared chiral object solution with R chirality or S chirality gradually into a prepared host solution with a preset concentration containing preset host molecules to obtain a mixed solution with saturated CD signals, then drawing a standard curve for marking the ee value of the chiral molecules based on successively adding standard object solutions of the chiral molecules with known ee values into the mixed solution and according to the continuously monitored CD signals of the mixed solution after the standard object solutions are added, and further solving a preset standard curve equation for calculating the ee value of the chiral molecules.
Further, step S500 includes:
step S501, detecting the saturation concentration of the CD signal of the mixed solution, and adding a standard object solution with a known ee value under the saturation concentration into the host solution;
step S502, reading a second CD signal intensity value of the subject solution added with the standard object solution at the preset position of the CD spectrum;
and S503, drawing a standard curve by taking the ee value of the standard object solution as a horizontal coordinate and the second CD signal intensity value as a vertical coordinate to obtain the preset standard curve equation.
It should be noted that, in this embodiment, in the process of using standard guest solutions with known ee values to draw a standard curve, so as to calculate a preset standard curve equation for calculating the ee value of the chiral molecule, all the used standard guest solutions need to satisfy the requirement of CD signal saturation.
In this embodiment, the predetermined position is any position between 250nm (nanometers) and 260nm on the CD spectrum, for example, 255 nm.
Specifically, for example, it is examined that a mixed solution saturated with CD signal is obtained by adding chiral guest solutions of R chirality to a prepared host solution having a concentration of 0.05mM in series, the saturated concentration of the chiral guest solution of R chirality is prepared, a standard guest solution of 1-phenylethyl alcohol of R chiral molecule having an ee value of + 100% is prepared in an aqueous solution according to the saturated concentration, then, the prepared standard guest solution of 1-phenylethyl alcohol is added to the prepared host solution having the concentration of 0.05mM in the same manner, a standard curve is plotted with the ee value of the standard guest solution of 1-phenylethyl alcohol added as abscissa and the CD signal intensity value at 255nm as ordinate on the CD spectrum of the mixed solution after the addition of the standard guest solution of 1-phenylethyl alcohol, and a standard curve equation y of 0.145x-0.443 is obtained, wherein, y is the CD signal intensity, and x is the ee value.
Further, a preferred embodiment for calculating the preset standard curve equation is proposed:
preparing standard object solutions of high-concentration chiral molecules with different ee values, adding the standard object solutions into a 0.05mM host solution, uniformly mixing to enable the concentration of the standard object solutions to reach the CD saturation concentration, and then measuring the CD spectra generated by the host solution induced by the standard object solutions with different ee values. The scanning interval of the CD spectrum is 240nm to 400nm, the CD spectrum is scanned for 3 times in total, and the spectrum is subjected to average treatment. And taking the ee value as an abscissa, and selecting the signal intensity value at 255nm of the CD spectrum as an ordinate to be used as a CD standard curve to obtain a corresponding standard equation.
And step S200, adding a guest solution of chiral molecules to be measured with saturated concentration into the host solution to obtain a solution to be detected with saturated circular dichroism spectrum CD signals.
Adding a guest solution of chiral molecules to be measured with saturated concentration into a prepared host solution with preset concentration and containing preset host molecules to obtain a solution to be detected with saturated CD signals.
Specifically, for example, in the process of detecting that a chiral guest solution of the R chirality is added successively to a prepared host solution with a concentration of 0.05mM to obtain a mixed solution with a saturated CD signal, the saturated concentration of the chiral guest solution of the R chirality is prepared in advance in an aqueous solution to be measured with a certain saturated concentration of a guest solution of a chiral molecule to be measured (unknown ee value), and then the guest solution of the chiral molecule to be measured is added to the prepared host solution with a concentration of 0.05mM to obtain a solution to be detected with a saturated CD signal.
In this embodiment, the guest solution of the chiral molecule to be measured is also prepared in advance based on an aqueous solution.
In the embodiment, the host solution containing the preset host molecules, the chiral guest solution with single chirality, the standard guest solution with known ee value and the guest solution of the chiral molecules to be measured are all prepared in the aqueous solution, so that the detection solvent of the invention is water, and compared with the traditional non-aqueous solvent, the detection solvent of the invention is green and environment-friendly, and the detection cost is reduced.
Further, in another embodiment, when a guest solution of the chiral molecule to be measured is added to a host solution containing a preset host molecule and having a preset concentration to prepare a solution to be detected with saturated CD signals, after the solution to be detected is sufficiently and uniformly mixed, the incubation time of the solution to be detected can be kept to be not more than 1 minute, or after the solution to be detected is sufficiently and uniformly mixed, the CD signal intensity value at a preset position can be directly measured to calculate the ee value of the chiral molecule to be measured. Therefore, when the solution to be detected for detecting the ee value of the chiral molecule to be detected is prepared, the measurement and calculation of the ee value of the chiral molecule to be detected can be carried out only by fully and uniformly mixing the mixed solution and the guest solution of the chiral molecule to be detected and keeping extremely short incubation time or waiting for incubation, so that the rapid measurement of the ee value is realized, the method for detecting the ee value of the chiral molecule is ensured, and the method is suitable for high-flux asymmetric catalytic condition screening and real-time monitoring of catalytic reaction.
And step S300, measuring a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate the ee value of the chiral molecule to be measured.
After a solution to be detected for detecting the ee value of the chiral molecule to be detected is prepared, a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum is determined based on the CD spectrum of the solution to be detected, and then the first CD signal intensity value is directly substituted into a preset standard curve equation for calculating the ee value of the chiral molecule to be detected, so that the ee value of the chiral molecule to be detected is calculated.
Specifically, for example, if the chiral molecule to be measured is 1-phenylethyl alcohol, after a guest solution with unknown ee value of 1-phenylethyl alcohol prepared in advance in an aqueous solution is added successively to the mixed solution with saturated CD signal to prepare a solution to be detected with saturated CD signal, based on the determination of the CD spectrum of the solution to be detected (determination 3 times), the CD signal intensity value of the solution to be detected at 255nm (average value of the CD signal intensity values at 255nm of the 3 times CD spectrum) is selected, then the CD signal intensity value at 255nm is substituted as "y" into a standard guest solution with 1-phenylethyl alcohol with known ee value added in advance in the mixed solution with saturated CD signal, with the ee value of the standard guest solution with 1-phenylethyl alcohol added as an abscissa, and (3) on a CD spectrum of a mixed solution added with the standard guest solution of the 1-phenethyl alcohol, taking a CD signal intensity value at 255nm as a vertical coordinate to draw a standard curve, and solving the obtained standard curve equation y to be 0.145x-0.443 to obtain the ee value of the chiral molecule 1-phenethyl alcohol to be measured in the asymmetric catalytic reaction.
In this example, a host solution of a predetermined concentration of a predetermined host molecule having a structure and a stereoisomer as shown in fig. 4 was prepared in an aqueous solution; adding a pre-prepared chiral object solution with R chirality or S chirality into a prepared host solution with a preset concentration and containing preset host molecules to saturate CD signals of the mixed solution so as to obtain the saturated concentration of the chiral object solution; respectively adding standard guest solutions of chiral molecules with known ee values of saturated concentrations into a host solution with preset concentration based on the data, and drawing a standard curve for marking the ee values of the chiral molecules according to detected CD signals of a mixed solution after the standard guest solutions are added, so as to calculate a preset standard curve equation for calculating the ee values of the chiral molecules; adding an object solution of chiral molecules to be measured with saturated concentration into a prepared host solution with preset concentration and containing preset host molecules to obtain a solution to be detected with saturated CD signals; and selecting a first CD signal intensity value of the solution to be detected at a preset position based on the CD spectrum of the solution to be detected, and directly substituting the first CD signal intensity value into a preset standard curve equation for calculating the ee value of the chiral molecule to calculate the ee value of the chiral molecule to be detected.
The method realizes that the ee value of the chiral molecules is measured by using the method for measuring the chiral molecule configuration by using the circular dichroism spectrum, and compared with the traditional method for measuring by using the chromatography, the method can more quickly, simply and conveniently detect the ee value of the chiral molecules, thereby being suitable for high-flux asymmetric catalytic condition screening and real-time monitoring of catalytic reaction.
Further, in another embodiment, the process of applying the chiral molecular ee value measurement method of the invention to real-time monitoring of asymmetric catalytic reaction may specifically be:
1mM R-1-phenylethanol acetate and 0.05mM macrocyclic host are added to a circular dichroism cell, followed by a final concentration of 100mM sodium hydroxide solution. And (3) placing the sample cell in a circular dichroism chromatograph, and testing the CD signal of the 255nm position of the sample in real time. The R-acetic acid-1-phenethyl alcohol ester is hydrolyzed under the alkaline condition to generate chiral R-1-phenethyl alcohol, and CD signals are changed. The real-time monitoring of the asymmetric catalytic reaction can be realized through the monitoring of the CD signal.
Further, in another embodiment, the ee value measurement method of chiral molecules of the present invention can be applied to monitor the racemization reaction specifically as follows:
a round bottom flask of 50m L was charged with a 25m L aqueous solution of 80mM R-1-phenylethyl alcohol to which was added 0.5g Amberlyst-15 (an acidic catalyst), heated at 65 ℃ under reflux with rapid stirring at intervals, the 25 μ L solution was mixed with a 2m L0.05.05 mM bulk solution and the CD spectrum was measured 3 times, after averaging the intensity at 255nm was substituted into a standard curve equation to calculate the ee value, allowing real-time monitoring of the racemization process.
Further, in another embodiment, the ee value measuring method for chiral molecules of the invention can be applied to the determination of optical purity of chiral drugs specifically as follows:
the chiral drug is prepared into an aqueous solution and then added into a main solution, and the CD signal value of the system is measured. And substituting the signal intensity value at 255nm into a standard curve equation to calculate the optical purity of the chiral drug.
Further, based on the above first embodiment of the method for measuring an ee value of a chiral molecule, a second embodiment of the method for measuring an ee value of a chiral molecule of the present invention is provided.
Referring to fig. 3, in the present embodiment, after the step S300, the method for measuring ee of chiral molecule further includes:
step S600, recovering the amide molecule tube from the solution to be detected for preparing a new main body solution.
The method comprises the steps of determining a first CD signal intensity value of a solution to be detected at a preset position based on a CD spectrum of the solution to be detected, substituting the first CD signal intensity value into a preset standard curve equation obtained through calculation in advance, calculating an ee value of the chiral molecule to be measured, and recovering preset main body molecules from the solution to be detected for preparing a new main body solution based on adjusting the pH value of the solution to be detected, so that the used main body molecules are recovered and reused.
Further, step S600 includes:
step S601, adjusting the pH value of the solution to be detected to obtain the amide molecular tube through precipitation;
step S602, the amide molecular tube obtained by precipitation is subjected to ultrasonic cleaning and drying treatment to complete the recovery of the amide molecular tube.
Specifically, for example, a first CD signal intensity value of a solution to be detected at a preset position of a CD spectrum is determined based on the CD spectrum of the solution to be detected, then the first CD signal intensity value is substituted into a preset standard curve equation obtained by calculation in advance, after an ee value of a chiral molecule to be measured is calculated, a hydrochloric acid adjusting solution is added into the solution to be detected (the solution after the ee value of the chiral molecule is measured) until the pH value of the solution is adjusted to about 1, so that an amide molecular tube in the solution to be detected is precipitated under an acidic condition, then the precipitated amide molecular tube is filtered and recovered, dichloromethane is added into the recovered amide molecular tube for ultrasonic cleaning, and after the cleaning is performed twice, the pure amide molecular tube is dried to be reserved for a subsequent repeated preparation of a main body solution.
In this embodiment, a first CD signal intensity value of the solution to be detected at a preset position is determined based on a CD spectrum of the solution to be detected, then the first CD signal intensity value is substituted into a preset standard curve equation obtained by calculation in advance, and after an ee value of the chiral molecule to be detected is calculated, an amide molecular tube is recovered from the solution to be detected for preparing a new main body solution based on adjustment of the ph of the solution to be detected, so that recycling of the used amide molecular tube is realized, and the detection cost is further reduced.
It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or system that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or system. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a process, method, article, or system that comprises the element.
The above-mentioned serial numbers of the embodiments of the present invention are merely for description and do not represent the merits of the embodiments.
The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. A method for measuring an ee value of an enantiomeric excess of a chiral molecule is characterized by comprising the following steps:
preparing a main body solution containing preset main body molecules;
adding a guest solution of chiral molecules to be measured with saturated concentration into the host solution to obtain a solution to be detected with saturated circular dichroism spectrum CD signals;
and measuring a first CD signal intensity value of the solution to be detected at a preset position of a CD spectrum, and substituting the first CD signal intensity value into a preset standard curve equation to calculate the ee value of the chiral molecule to be detected.
2. The method for measuring ee of a chiral molecule according to claim 1, wherein said step of preparing a host solution containing a predetermined host molecule is followed by the steps of:
adding a single chiral object solution into the host solution to obtain a mixed solution saturated with CD signals;
and drawing the preset standard curve equation based on the mixed solution and the standard object solution with the known ee value.
3. The method for measuring ee of chiral molecule according to claim 2, wherein said step of plotting said predetermined standard curve equation based on said mixed solution and a standard guest solution of known ee comprises:
detecting the saturated concentration of the CD signal of the mixed solution, and adding a standard guest solution with a known ee value under the saturated concentration into the host solution;
reading a second CD signal intensity value of the host solution added with the standard guest solution at the preset position of the CD spectrum;
and drawing a standard curve by taking the ee value of the standard object solution as a horizontal coordinate and the second CD signal intensity value as a vertical coordinate so as to obtain the preset standard curve equation.
4. The method of claim 2, wherein the host solution, the chiral guest solution and the standard guest solution are prepared in an aqueous solution.
5. The method of claim 2, wherein the chiral guest solution of single chirality comprises a chiral guest solution of R chirality and a chiral guest solution of S chirality.
6. The method for measuring ee of a chiral molecule according to any one of claims 1 to 3, wherein said predetermined host molecule is an amide molecule tube.
7. The method for measuring ee of chiral molecule according to claim 6, further comprising, after said steps of determining a first CD signal intensity value of said solution to be detected at a predetermined position of CD spectrum, and substituting said first CD signal intensity value into a predetermined standard curve equation to calculate ee of said chiral molecule to be measured:
recovering the amide molecule tube from the solution to be detected for preparing a new main solution.
8. The method for measuring ee of chiral molecules according to claim 7, wherein said step of recovering said tube of amide molecules from said solution to be detected comprises:
adjusting the pH value of the solution to be detected to precipitate to obtain the amide molecular tube;
and carrying out ultrasonic cleaning and drying treatment on the amide molecular tube obtained by precipitation so as to complete the recovery of the amide molecular tube.
9. The method for measuring ee of chiral molecules according to claim 1, wherein the concentration of said host solution is between 0.01mM and 0.1 mM.
10. The method for measuring ee of chiral molecules according to claim 1, wherein the predetermined position is between 250nm and 260nm of the CD spectrum.
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| WO2020056012A1 (en) * | 2018-09-11 | 2020-03-19 | Georgetown University | Quantitative auxiliary-free chirality sensing with a metal probe |
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