Application and analysis method of cyclodextrin in the analysis of catecholamine molecular isomers

Abstract

The invention relates to the field of analysis and testing, and in particular to an application of cyclodextrin in analyzing catecholamine molecular isomers and an analysis method thereof; the steps include: S1, mixing a sample to be tested with cyclodextrin, and non-covalently combining the catecholamine in the sample to be tested with the cyclodextrin to obtain a mixed solution; S2, treating the mixed solution with an ionizing device to generate composite gas-phase ions, and then measuring the ion mobility or ion collision cross section of the composite gas-phase ions to distinguish the isomers of catecholamines; the analysis method of the invention overcomes the difficulty that mass spectrometry, spectral analysis and other methods cannot analyze and identify the molecular types of catecholamine isomers, and compared with traditional isomer molecular type analysis techniques and methods, the method is simple, rapid and accurate, and can analyze each isomer in the mixed sample and its content; the analysis method of the invention requires a small amount of sample and has high sensitivity, and has broad application prospects in the fields of pharmaceuticals, clinical medical testing, chemical industry, etc.

Description

Technical Field

[0001] The invention relates to the field of analysis and testing, and in particular to the application of cyclodextrin in analyzing catecholamine molecular isomers and an analysis method. Background Art

[0002] Catecholamines are an important class of biologically active molecules and important neurotransmitters in the body. They have a wide range of physiological effects, such as exciting nerve receptors, stimulating vasoconstriction, and increasing cardiac output and heart rate. Catecholamines are also a commonly used clinical therapeutic drug. Commonly used catecholamine drugs in clinical practice include dopamine, epinephrine, norepinephrine, etc. Their chemical composition and molecular structure are as follows: Figure 1 As shown, it can be seen that each catecholamine has a different molecular structure, or even different chiral isomers, which exhibit different physiological functions in clinical practice, such as:

[0003] 1. Dopamine, its clinical manifestations are that when used in small doses, it can dilate blood vessels and increase urine volume, etc., when used in medium doses, it can increase cardiac output and raise systolic blood pressure, and when used in large doses, it has a significant effect of raising systolic and diastolic blood pressure.

[0004] 2. Epinephrine has the clinical manifestation of contracting the skin, mucous membranes, and visceral blood vessels, causing coronary artery dilation, increased heart rate, and relaxation of bronchial and gastrointestinal smooth muscles. It is mainly used clinically to treat diseases such as bronchial asthma, anaphylactic shock, hypotension, and cardiac arrest.

[0005] 3. Norepinephrine has the effect of causing vasoconstriction, increasing systolic and diastolic blood pressure, and reflexively slowing the heart rate. It is mainly used clinically to treat hypotension and hypovolemia symptoms caused by acute myocardial infarction, as well as to maintain blood pressure after cardiac arrest resuscitation.

[0006] Due to the different clinical effects of different catecholamine drugs, improper selection and use can cause serious side effects, such as local tissue necrosis and renal failure, aggravation of heart failure, induction of acute coronary syndrome, and increased blood pressure and blood sugar, which have the opposite effect of the therapeutic effect. Therefore, whether in the fields of drug research and development and production, clinical medicine, etc., accurate analysis and identification of catecholamine types and isomer structure types, and the content of various catecholamine isomers have important scientific significance and application value.

[0007] At present, most catecholamine drugs used in clinical practice usually exist in the form of racemates, that is, two isomers of different chirality exist in the drug at the same time, that is, the catecholamine content of the two chiral isomers is almost relative. However, since the biological effects and effects of chiral drug molecules are closely related to the chiral environment of molecules such as enzymes, receptors and antibodies in animals, the use of racemic drugs may lead to incorrect pharmacokinetic behavior and effects. For example, clinical results show that the L-enantiomers of adrenaline and norepinephrine have stronger physiological activity than their D-enantiomers; L-dopamine has clear pharmacological activity in the treatment of neurodegenerative diseases, but the therapeutic effect of D-dopamine is not obvious and may even produce adverse reactions. Therefore, the accurate identification and separation of different catecholamine chiral isomers are of great significance in drug development and pharmaceuticals, clinical medicine, etc.

[0008] The results of drug development and clinical use show that the biological function of a drug is closely related to the chemical composition and molecular structure of the drug molecule. For analytical chemistry, due to the same molecular composition and similar molecular structure, the analysis and identification of molecular isomers is often very difficult, especially for the analysis and identification of small molecule chiral isomers. This is because the chemical properties or physical properties of different isomers are often very similar, such as having exactly the same molecular mass, which makes mass spectrometry analysis powerless, and the chiral structure of the molecule makes it impossible for conventional mass spectrometry, chromatography, and spectral analysis methods to distinguish its chiral species. In the past few decades, molecular structure analysis has been one of the research goals of analytical scientists, and the structural analysis of molecular isomers is one of the difficulties. This is because the molecules of isomers have exactly the same molecular mass, so they cannot be directly analyzed by mass spectrometry. Some isomers have very small structural differences, such as chiral isomers, and general spectroscopy and nuclear magnetic resonance cannot distinguish them. Summary of the invention

[0009] The purpose of the present invention is to provide an analytical method for high-sensitivity, rapid and accurate analysis and identification of several catecholamine isomer molecules in view of the deficiencies in the prior art.

[0010] To achieve the above object, the technical solution adopted by the present invention is:

[0011] The first aspect of the present invention is to provide a method for analyzing catecholamine molecular isomers using cyclodextrin, the steps comprising:

[0012] S1, mixing the sample to be tested with cyclodextrin, and the catecholamine in the sample to be tested is non-covalently bound to the cyclodextrin to prepare a mixed solution;

[0013] S2. Treating the mixed solution with an ionization device to generate complex gas-phase ions, and then measuring the ion mobility or ion collision cross section of the complex gas-phase ions to distinguish the isomers of catecholamines.

[0014] Preferably, in step S1, the cyclodextrin includes at least one of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, α-cyclodextrin derivatives, β-cyclodextrin derivatives, and γ-cyclodextrin derivatives.

[0015] Preferably, in step S1, the catecholamines include at least one of L-epinephrine, D-epinephrine, L-norepinephrine, D-norepinephrine, L-dopamine, and D-dopamine.

[0016] Preferably, in step S1, the sample to be tested includes: medicine, blood or urine.

[0017] Preferably, in step S2, the composite gas phase ions include: [LE / DE+αCD / βCD / γCD+H] + , [LD / DD+αCD / βCD / γCD+H] + , [LNE / DNE+αCD / βCD / γCD+H] + At least one of .

[0018] Preferably, in step S2, the ionization device comprises: an electrospray ionization source, a laser-assisted desorption ionization source, and a desorption electrospray ionization source.

[0019] The second aspect of the present invention is to provide the use of cyclodextrin in analyzing catecholamine molecular isomers.

[0020] Preferably, the cyclodextrin includes at least one of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, α-cyclodextrin derivatives, β-cyclodextrin derivatives and γ-cyclodextrin derivatives.

[0021] Preferably, the catecholamines include at least one of L-type adrenaline, D-type adrenaline, L-type norepinephrine, D-type norepinephrine, L-type dopamine, and D-type dopamine.

[0022] The present invention adopts the above technical solution, and has the following technical effects compared with the prior art:

[0023] The analysis method of the present invention overcomes the difficulty that mass spectrometry, spectral analysis and other methods cannot analyze and identify the types of catecholamine isomer molecules. Compared with traditional isomer molecule type analysis techniques and methods, the method is simple, rapid and accurate, and can analyze each isomer and its content in a mixed sample; the analysis method of the present invention requires a small amount of sample and has high sensitivity, and has broad application prospects in the fields of pharmaceuticals, clinical medical testing, chemical industry, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The diagrams are the types and chemical structures of catecholamine molecules and the molecular structure of cyclodextrin; wherein a is the enantiomer of catecholamine; and b is the cyclodextrin (CD) molecule.

[0025] Figure 2 The results are the mass spectrometry analysis of several catecholamine-cyclohexine molecular mixed solutions and the ion mobility spectrometry analysis of gas phase complex ions; where a is the IMS spectrum of the binary complex of adrenaline enantiomers (LE: L-adrenaline, DE: D-adrenaline) and CD, including [LE / DE+α / β / γ-CD+H] + ; b is the IMS spectrum of norepinephrine enantiomers (LNE: L-norepinephrine, DNE: D-norepinephrine) and CD binary complexes, including [LNE / DNE+α / β / γ-CD+H] + ; c is the IMS spectrum of the binary complex of dopa enantiomers (LD: L-dopamine, DD: D-dopamine) and CD, including [LD / DD+α / β / γ-CD+H] + ; The IMS containing the L-type chiral isomer complex is marked in blue, and the IMS containing the D-type chiral isomer complex is marked in red; the mass-to-charge ratio of the complex ion is expressed in m / z, and the Rp-p value represents the resolution of the ion mobility spectra. DETAILED DESCRIPTION

[0026] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0028] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but they are not intended to limit the present invention.

[0029] Example

[0030] This embodiment provides an analytical method for analyzing catecholamine molecular isomers using cyclodextrin, the steps comprising:

[0031] S1. Weigh the sample to be tested and cyclodextrin, dissolve them in ultrapure water respectively and mix them, so that the catecholamine in the sample to be tested is non-covalently bound to the cyclodextrin to prepare a mixed solution (the final solution of catecholamine and cyclodextrin is about 10 μM);

[0032] S2. The mixed solution is treated with an electrospray ionization-trapped ion mobility spectrometry time-of-flight mass spectrometer (TIMS TOF MS, Bruker Daltonik, Germany) to generate complex gas-phase ions, and then the ion mobility or ion collision cross section of the complex gas-phase ions is measured. The types of catecholamine enantiomers contained in the above-mentioned complex ions are obtained by analyzing and calculating the ion mobility or ion collision cross section of each ion in the IMS spectrum.

[0033] The analysis results are as follows Figure 2 As shown, different catecholamine isomers are clearly distinguished.

[0034] In summary, the analysis method of the present invention overcomes the difficulty that mass spectrometry, spectral analysis and other methods cannot analyze and identify the types of catecholamine isomer molecules. Compared with traditional isomer molecular type analysis techniques and methods, the method is simple, rapid and accurate, and can analyze each isomer and its content in a mixed sample; the analysis method of the present invention requires a small amount of sample and has high sensitivity, and has broad application prospects in the fields of pharmaceuticals, clinical medical testing, chemical industry, etc.

[0035] The above description is only a preferred embodiment of the present invention, and does not limit the implementation mode and protection scope of the present invention. For those skilled in the art, it should be aware that all solutions obtained by equivalent substitutions and obvious changes made using the description and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for analyzing catecholamine molecular isomers using cyclodextrin, characterized in that the steps include: S1, mixing the sample to be tested with cyclodextrin, and non-covalently binding the catecholamine in the sample to be tested with the cyclodextrin to obtain a mixed solution; S2. Treating the mixed solution with an ionization device to generate complex gas-phase ions, and then measuring the ion mobility or ion collision cross section of the complex gas-phase ions to distinguish the isomers of catecholamines.

2. The analysis method according to claim 1, characterized in that In step S1, the cyclodextrin includes at least one of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, α-cyclodextrin derivatives, β-cyclodextrin derivatives, and γ-cyclodextrin derivatives.

3. The analysis method according to claim 1, characterized in that In step S1, the catecholamines include at least one of L-epinephrine, D-epinephrine, L-norepinephrine, D-norepinephrine, L-dopamine, and D-dopamine.

4. The analysis method according to claim 1, characterized in that In step S1, the sample to be tested includes: medicine, blood or urine.

5. The analysis method according to claim 1, characterized in that In step S2, the composite gas phase ions include: [LE / DE+αCD / βCD / γCD+H] + , [LD / DD+αCD / βCD / γCD+H] + , [LNE / DNE+αCD / βCD / γCD+H] + At least one of .

6. The analysis method according to claim 1, characterized in that In step S2, the ionization device includes: an electrospray ionization source, a laser-assisted desorption ionization source, and a desorption electrospray ionization source.

7. Application of cyclodextrin in the analysis of catecholamine molecular isomers.

8. The use according to claim 7, characterized in that: Cyclodextrins include: At least one of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, α-cyclodextrin derivatives, β-cyclodextrin derivatives, and γ-cyclodextrin derivatives.

9. The use according to claim 7, characterized in that: The catecholamines include at least one of L-type adrenaline, D-type adrenaline, L-type norepinephrine, D-type norepinephrine, L-type dopamine, and D-type dopamine.

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