Rotatable chromogenic functional molecule as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of functional food detection, and provides a rotatable chromogenic functional molecule as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing a coenzyme derivative solution, an alkaline compound solution and a crocetin solution, and carrying out conjugate coupling reaction to obtain the rotatable chromogenic functional molecule. The method is green, environment-friendly, easy to operate and relatively high in yield; the obtained rotatable chromogenic functional molecule has the advantages of high sensitivity coefficient, good pH stability, good solvent polarity tolerance, excellent light stability and low detection lower limit, and can efficiently, accurately and visually measure the viscosity of the liquid syrup fluid in situ.

Description

Technical Field

[0001] The present invention relates to the technical field of functional food detection, and in particular to a rotatable chromogenic functional molecule and a preparation method and application thereof. Background Art

[0002] Most liquid syrup fluid products are high-sugar drinks extracted from natural plant juices. They contain a variety of minerals, organic acids, and ingredients such as calcium and magnesium, and have both high nutritional value and application value. The viscosity of liquid syrup water refers to its viscosity, which is closely related to taste and mellowness. Taking maple syrup water as an example, maple syrup water can present different consistencies and colors through blending, which is directly related to the grade classification of the final product. High-grade maple syrup water has higher viscosity, rich and mellow taste, poor fluidity, and a fuller mellowness as a whole, and the solid content of internal nutrients is also higher. In contrast, low-grade maple syrup water has lower viscosity, good fluidity, and is smoother to swallow, but has a poor taste, lower mellowness, and a relatively low solid content of internal nutrients. It can be seen that the precise control of the consistency is extremely critical. It not only reflects the grade difference, but also an intuitive reflection of the content of multiple components. Specifically, light-colored maple syrup water has a relatively light taste, is transparent and has good fluidity, and is generally suitable for breakfast, afternoon tea or making healthy drinks; medium-colored maple syrup has an improved taste, which can give pastries, cakes and bread a unique aroma and flavor, and is therefore often used to make such foods; dark-colored maple syrup has a richer taste and aroma, which can make food more condensed and formable, and have a better texture, and is generally used to make strong-flavored desserts, barbecue, scones and cooking seasonings.

[0003] One of the key quality indicators of liquid syrup fluid products is viscosity. For low-viscosity syrup-like liquid fluids, the detection accuracy of existing detection tools is insufficient. For example, when measuring low-viscosity fluids, the traditional falling ball, capillary, and vibration viscometers have relatively weak measurement signals due to the small resistance of the fluid to the rotor, and are easily affected by various factors such as the mechanical error of the instrument itself and environmental vibration, resulting in large deviations in the error of the measurement results, making it difficult to effectively measure the viscosity of low-viscosity syrup fluids. In contrast, although the rotational viscometer can effectively measure low-viscosity syrup liquids by switching the viscosity sensing mechanical component, the relative force of the fluid is mainly used to sense the mechanical stirring process, and the micro-area environment of the object to be tested will be destroyed during the detection process, which belongs to shear destructive detection, which will distort the measurement results and make it difficult to obtain accurate and close values ​​of the actual viscosity of the micro-environment. In actual applications, many problems are also exposed. The amount of sample required for the test is large, and the entire test process is highly dependent on the equipment, which is not only time-consuming, but also consumes a lot of manpower.

[0004] Therefore, there is an urgent need to develop a rotatable chromogenic functional molecular measurement tool that can achieve accurate, in situ measurement, high efficiency, ease of operation, and visual detection, as well as its preparation method and application. Summary of the invention

[0005] In view of this, the present invention provides a rotatable chromogenic functional molecule and a preparation method and application thereof, so as to solve the problem that the viscosity of liquid syrup fluid is difficult to be accurately measured by existing detection tools and the measurement process is complicated and inefficient.

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

[0007] The present invention provides a rotatable chromogenic functional molecule, the structural formula of the rotatable chromogenic functional molecule is shown in Formula I:

[0008]

[0009] Wherein, the Y is a coenzyme derivative group, and the coenzyme derivative group includes

[0010]

[0011] The present invention also provides a method for preparing a rotatable chromogenic functional molecule, comprising the following steps:

[0012] The coenzyme derivative solution, the alkaline compound solution and the saffron aldehyde solution are mixed to carry out a conjugated coupling reaction to obtain a rotatable color-forming functional molecule; wherein the coenzyme derivative in the coenzyme derivative solution includes

[0013]

[0014] Preferably, the molar ratio of the coenzyme derivative in the coenzyme derivative solution, the alkaline compound in the alkaline compound solution and the saffron aldehyde in the saffron aldehyde solution is 1:1-10:1-5.

[0015] Preferably, the temperature of the conjugation coupling reaction is 40 to 120° C., and the time is 1 to 32 hours.

[0016] Preferably, the alkaline compound in the alkaline compound solution includes one or more of sodium carbonate, cesium carbonate, aluminum hydroxide, sodium bicarbonate, potassium carbonate, potassium bicarbonate, triethylamine, trimethylamine, lithium bistrimethylsilylamide, lithium diisopropylamide, sodium bis(trimethylsilyl)amide, potassium bis(trimethylsilyl)amide, imidazole, pyridine, ammonia water and N,N,N',N'-tetramethylethylenediamine.

[0017] Preferably, the solvent in the coenzyme derivative solution, the solvent in the alkaline compound solution and the solvent in the saffron aldehyde solution independently include one or more of acetonitrile, tetrahydrofuran, N,N-dimethylformamide, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, propanol, n-butanol, ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol and 1,3-butanediol.

[0018] Preferably, the molar concentration of the coenzyme derivative solution is 1 to 3 mol / L, the molar concentration of the alkaline compound solution is 1 to 10 mol / L, and the molar concentration of the saffron aldehyde solution is 1 to 5 mol / L.

[0019] The present invention also provides an application of a rotatable chromogenic functional molecule prepared by the preparation method of the rotatable chromogenic functional molecule in detecting the viscosity of a liquid syrup fluid.

[0020] It can be seen from the above technical solution that compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. The rotatable chromogenic functional molecule of the present invention shows extremely high sensitivity to viscosity, and the viscosity sensitivity coefficient can be as high as 0.78, which can realize accurate, in-situ, and visual detection of the viscosity of maple syrup water containing pseudoplastic fluid components. At the same time, the rotatable chromogenic functional molecule of the present invention has a stable chemical structure, and can exist stably for a long time even in maple syrup water with complex ingredients. Moreover, the color wavelength peak of this rotatable chromogenic functional molecule is 510nm, showing a typical green light, bright color, and excellent visualization effect. In addition, the rotatable chromogenic functional molecular tool of the present invention can release stable light signals within a wide pH range, and can still maintain the signal well even if it is excited by an external light source for a long time, and has excellent solvent tolerance.

[0022] 2. The rotatable chromogenic functional molecule of the present invention is formed by conjugating and coupling a coenzyme derivative and a saffron extract, saffron aldehyde. The preparation method is simple, the yield is high, the raw material source is rich and the cost is low. At the same time, the rotatable chromogenic functional molecule of the present invention has the characteristics of low carbon, environmental protection and degradability. When the rotatable chromogenic functional molecule is used to measure the viscosity of maple syrup water, the use cost is as low as milligram level, and the measurement process is also simple. It can efficiently, accurately, in situ and visualize the measurement of the viscosity of liquid syrup fluid, and is suitable for large-scale preparation and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0024] Figure 1 Schematic diagram of the mechanism of testing the viscosity of maple syrup water micro-areas using the rotatable chromogenic functional molecules prepared in Example 1;

[0025] Figure 2 This is the mass spectrum of the rotatable chromogenic functional molecule prepared in Example 1;

[0026] Figure 3 This is the nuclear magnetic resonance spectrum of the rotatable chromogenic functional molecule prepared in Example 1;

[0027] Figure 4 This is a fluorescence intensity diagram of the rotatable chromogenic functional molecule prepared in Example 1 in different maple syrup waters;

[0028] Figure 5 Spectra of the rotatable chromogenic functional molecules prepared in Example 1 in solutions with different viscosities;

[0029] Figure 6 This is a linear fitting diagram between the optical signal intensity and viscosity of the rotatable chromogenic functional molecule prepared in Example 1;

[0030] Figure 7 This is a graph showing the photostability test results of the rotatable chromogenic functional molecule prepared in Example 1 in glycerol and purified water;

[0031] Figure 8 The spectra of the rotatable chromogenic functional molecules prepared in Example 1 in different pH atmospheres;

[0032] Fig. 9 The absorption spectra of the rotatable chromogenic functional molecules prepared in Example 1 in different solvents;

[0033] Fig.10 This is a graph showing the detection limit test results of the rotatable chromogenic functional molecule prepared in Example 1. DETAILED DESCRIPTION

[0034] The present invention provides a rotatable chromogenic functional molecule, the structural formula of the rotatable chromogenic functional molecule is shown in Formula I:

[0035]

[0036] Wherein, the Y is a coenzyme derivative group, and the coenzyme derivative group includes

[0037]

[0038] The present invention also provides a method for preparing a rotatable chromogenic functional molecule, comprising the following steps:

[0039] The coenzyme derivative solution, the alkaline compound solution and the safranal solution are mixed to carry out a conjugated coupling reaction to obtain a rotatable color-producing functional molecule (CoenQX-Saf);

[0040] Wherein, the coenzyme derivative in the coenzyme derivative solution includes

[0041] In the present invention, the mixing is preferably performed by first mixing the coenzyme derivative solution and the alkaline compound solution, heating them, and then adding the saffron aldehyde solution after heating to the conjugated coupling reaction temperature; the stirring rate of the mixing is preferably 1000-2200rpm, more preferably 1200-2000rpm, and more preferably 1600-1800rpm; the heating rate of the heating is preferably 1°C / min-10°C / h, more preferably 50°C / h-20°C / h, and more preferably 30°C / h; the addition rate of the saffron aldehyde solution is preferably 1-10mL / min, more preferably 2-8mL / min, and more preferably 3-5mL / min.

[0042] In the present invention, the molar ratio of the coenzyme derivative in the coenzyme derivative solution, the alkaline compound in the alkaline compound solution and the saffron aldehyde in the saffron aldehyde solution is 1:1-10:1-5, preferably 1:2-8:2-4, and more preferably 1:3-6:3-4.

[0043] In the present invention, the temperature of the conjugated coupling reaction is 40 to 120°C, preferably 50 to 100°C, more preferably 60 to 90°C, and more preferably 70 to 80°C; the time of the conjugated coupling reaction is 1 to 32h, preferably 5 to 28h, more preferably 10 to 20h, and more preferably 12 to 16h.

[0044] In the present invention, the alkaline compound in the alkaline compound solution includes one or more of sodium carbonate, cesium carbonate, aluminum hydroxide, sodium bicarbonate, potassium carbonate, potassium bicarbonate, triethylamine, trimethylamine, lithium bistrimethylsilylamide, lithium diisopropylamide, sodium bis(trimethylsilyl)amide, potassium bis(trimethylsilyl)amide, imidazole, pyridine, ammonia water and N,N,N',N'-tetramethylethylenediamine.

[0045] In the present invention, the solvent in the coenzyme derivative solution, the solvent in the alkaline compound solution and the solvent in the saffron aldehyde solution independently include one or more of acetonitrile, tetrahydrofuran, N,N-dimethylformamide, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, propanol, n-butanol, ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol and 1,3-butanediol.

[0046] In the present invention, the molar concentration of the coenzyme derivative solution is 1-3 mol / L, preferably 2 mol / L; the molar concentration of the alkaline compound solution is 1-10 mol / L, preferably 2-8 mol / L, further preferably 3-6 mol / L, more preferably 4-5 mol / L; the molar concentration of the saffron aldehyde solution is 1-5 mol / L, preferably 2-4 mol / L, more preferably 3 mol / L.

[0047] In the present invention, the method for preparing the coenzyme derivative solution is preferably mixing the coenzyme derivative with a solvent and stirring to obtain a coenzyme series derivative solution; the method for preparing the alkaline compound solution is preferably mixing the alkaline compound with a solvent and stirring to obtain an alkaline compound solution; the method for preparing the saffron aldehyde solution is preferably mixing saffron aldehyde with a solvent and stirring to obtain a saffron aldehyde solution.

[0048] In the present invention, the stirring temperature when preparing the coenzyme derivative solution, the alkaline compound solution and the saffron aldehyde solution is independently preferably room temperature 25°C; the stirring rate is independently preferably 100-1000rpm, further preferably 200-800rpm, more preferably 300-500rpm; the stirring time is independently preferably 1-12h, further preferably 2-10h, more preferably 3-6h.

[0049] In the present invention, the method for preparing the rotatable chromogenic functional molecule preferably further comprises the steps of extracting, purifying, centrifuging, crystallizing and drying the obtained rotatable chromogenic functional molecule in sequence.

[0050] In the present invention, the extraction is preferably performed by first reducing the pressure on the rotatable chromogenic functional molecules to remove the solvent, and then performing the extraction; the pressure of the reduced pressure is preferably -0.09 to -0.06 MPa, and more preferably -0.08 to -0.07 MPa; the extraction solution is preferably a mixed solution of dichloromethane and purified water; the volume ratio of dichloromethane to purified water is preferably 1 to 15:1, more preferably 2 to 13:1, and more preferably 5 to 10:1.

[0051] In the present invention, the purification is preferably to first remove the solvent from the rotatable chromogenic functional molecules obtained by extraction under reduced pressure, and then place the rotatable chromogenic functional molecules from which the solvent has been removed in a silica gel chromatographic column filled with silica gel powder for purification; the pressure of the reduced pressure is preferably -0.09 to -0.06 MPa, and more preferably -0.08 to -0.07 MPa; the mesh number of the silica gel powder is preferably 200 to 1000 mesh, further preferably 300 to 800 mesh, and more preferably 500 to 700 mesh; the solvent for the purification is preferably a mixed solvent of dichloromethane and methanol; the volume ratio of dichloromethane to methanol is preferably 1 to 100:1, further preferably 5 to 80:1, and more preferably 10 to 50:1.

[0052] In the present invention, the centrifugation is preferably to first reduce the pressure on the purified rotatable chromogenic functional molecules to remove the solvent, and then place the rotatable chromogenic functional molecules from which the solvent has been removed in a centrifuge for centrifugal treatment; the pressure of the reduced pressure is preferably -0.09 to -0.06 MPa, more preferably -0.08 to -0.07 MPa; the solution for the centrifugal treatment is preferably a mixed solution of ethanol and purified water; the volume ratio of ethanol to purified water is preferably 1 to 15:1, more preferably 2 to 12:1, more preferably 5 to 10:1; the centrifugal treatment The solid content is preferably 1-20 mg / mL, more preferably 5-18 mg / mL, and more preferably 10-12 mg / mL; the centrifugal treatment rate is preferably 2000-12000 r / min, more preferably 3000-10000 r / min, and more preferably 5000-8000 r / min; the centrifugal treatment time is preferably 0.1-2 h, more preferably 0.5-1.5 h, and more preferably 0.8-1.1 h; the number of centrifugal treatments is preferably 1-4 times, and more preferably 2-3 times.

[0053] In the present invention, the crystallization is preferably performed by first dispersing the rotatable chromogenic functional molecules obtained by centrifugation in a solution, performing static crystallization, and then vacuum filtering the rotatable chromogenic functional molecules obtained by crystallization; the solution is preferably a mixed solution of ethanol and purified water; the volume ratio of the ethanol and purified water is preferably 1:1-30, more preferably 1:5-25, and more preferably 1:10-20; the temperature of the static crystallization is preferably 1-10°C, more preferably 2-8°C, and more preferably 3-5°C; the time of the static crystallization is preferably 1-24h, more preferably 3-20h, and more preferably 5-15h; the solid content of the static crystallization is preferably 5-6 0mg / mL, further preferably 10~50mg / mL, more preferably 20~35mg / mL; the pressure of the vacuum filtration is preferably -0.09~-0.06MPa, more preferably -0.08~-0.07MPa; the filter paper for the vacuum filtration is preferably slow filter paper; the number of layers of the slow filter paper is preferably 1~3 layers, more preferably 2 layers; the washing solution for the vacuum filtration is preferably a mixed solution of ethanol and purified water; the volume ratio of ethanol and purified water is preferably 1:1~30, further preferably 1:5~25, more preferably 1:10~20; the number of washing times for the vacuum filtration is preferably 1~4 times, more preferably 2~3 times.

[0054] In the present invention, the drying is preferably carried out by placing the rotatable chromogenic functional molecules that have been filtered under reduced pressure in a freeze dryer for drying; the drying temperature is preferably -50 to -5°C, further preferably -45 to -10°C, and more preferably -40 to -25°C; the drying time is preferably 1 to 36 hours, further preferably 5 to 30 hours, and more preferably 10 to 18 hours.

[0055] In the present invention, the chemical structure of the rotatable chromogenic functional molecule contains rotatable conjugated single and double bonds, which can present different rotation states in maple syrup water with different micro-region viscosities, thereby releasing different apparent light signals; in high-viscosity maple syrup water, it mainly releases light signals in the form of radiation transition, showing a strong fluorescence signal, and in low-viscosity maple syrup water, it mainly dissipates excited state energy in the form of mechanical rotation, and the fluorescence signal is relatively weak or even invisible.

[0056] The present invention also provides an application of a rotatable chromogenic functional molecule prepared by the preparation method of the rotatable chromogenic functional molecule in detecting the viscosity of a liquid syrup fluid.

[0057] In the present invention, the viscosity of the liquid syrup fluid is preferably 1 cP to 1000 cP.

[0058] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0059] Example 1

[0060] 1. Dissolve 300.44 g (molar weight is 2 mol) of saffron aldehyde in acetonitrile, stir evenly at room temperature (25°C), the stirring rate is 500 rpm, the stirring time is 6 hours, and a 2 mol / L saffron aldehyde solution is obtained;

[0061] 2. 318.41 g (1 mol) of coenzyme derivative Dissolve in methanol, stir evenly at room temperature (25°C) at a stirring rate of 500 rpm for 6 h to obtain a 1 mol / L coenzyme derivative solution;

[0062] 3. Disperse 651.64 g (molar weight is 2 mol) of cesium carbonate in acetonitrile, stir thoroughly at room temperature (25°C), at a stirring rate of 500 rpm, for 6 hours, to obtain a 2 mol / L cesium carbonate solution;

[0063] 4. Directly mix the coenzyme derivative solution obtained in step 2 and step 3 and the cesium carbonate solution, heat up directly after mixing, stir while heating, the heating rate is 30°C / h, the stirring rate is 1600rpm, after heating to 80°C, add the saffron aldehyde solution obtained in step 1 to carry out conjugated coupling reaction, wherein the addition rate of the saffron aldehyde solution is 5mL / min, the reaction time is 16h, and after the reaction is completed, a rotatable color-forming functional molecule crude product is obtained;

[0064] 5. After the crude product of the rotatable chromogenic functional molecule obtained in step 4 is cooled to room temperature (25°C), the solvent is removed under a pressure of -0.09MPa; then, a mixed solution of dichloromethane and purified water in a volume ratio of 10:1 is used to extract the crude product of the rotatable chromogenic functional molecule from which the solvent is removed, and the organic phase is collected; then, the rotatable chromogenic functional molecule obtained by extraction is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a silica gel column filled with 600 mesh silica gel powder for purification, wherein the purification solvent is a mixed solvent of dichloromethane and methanol in a volume ratio of 50:1; then, the purified rotatable chromogenic functional molecule is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a centrifuge for centrifugal treatment twice, wherein each centrifugal treatment has a solution volume ratio of 1:1. A mixed solution of ethanol and purified water in a volume ratio of 0:1, a solid content of each centrifuge tube is 10 mg / mL, the centrifugal speed is 8000 r / min, and the centrifugal time is 1 hour; then the rotatable chromogenic functional molecules obtained by centrifugation are dispersed in a mixed solution of ethanol and purified water in a volume ratio of 1:15, and crystallized at a temperature of 5°C for 12 hours and the solid content of the crystals is controlled to be 30 mg / mL, and then the rotatable chromogenic functional molecules obtained by crystallization are subjected to vacuum filtration at a pressure of -0.09 MPa, wherein the filter paper for vacuum filtration is a 2-layer slow filter paper, and the mixed solution of ethanol and purified water in a volume ratio of 1:15 is used for washing twice during the filtration process; finally, the rotatable chromogenic functional molecules after vacuum filtration are placed in a freeze dryer at a temperature of -25°C for drying for 18 hours to obtain a light yellow solid powder of rotatable chromogenic functional molecules, referred to as CoenQ2-Saf.

[0065] After testing, the yield of the rotatable chromogenic functional molecule obtained in this example was 415.0 g, and the yield was 92.1%.

[0066] The schematic diagram of the mechanism of the rotatable chromogenic functional molecules prepared in this example on the viscosity of maple syrup water micro-regions is shown in FIG. Figure 1 As shown, from Figure 1 It can be seen that the rotatable chromogenic functional molecule CoenQ2-Saf can rotate freely in low-viscosity maple syrup water, and the energy it absorbs is dissipated through mechanical rotation, making it difficult to release a fluorescent signal. As the viscosity of the maple syrup water increases, its chemical structure is difficult to rotate, and the absorbed energy can only be dissipated through radiation transition, releasing a strong fluorescent signal, and thus the relative size of the maple syrup water viscosity can be obtained.

[0067] The relative molecular mass of the rotatable chromogenic functional molecule prepared in this example was analyzed, and the high-resolution mass spectrometry results are as follows: Figure 2 As shown, from Figure 2It can be seen that the relative molecular mass of the rotatable color-forming functional molecular tool of this embodiment is 450.62138 [M] + , molecular formula is C 29 H 38 O 4 The theoretical relative mass estimation value is 450.61, that is, in terms of relative molecular mass, it can be found that the product obtained in this embodiment is consistent with the target product; in addition, the chemical structure of the rotatable chromogenic functional molecule prepared in this embodiment is also confirmed, and the nuclear magnetic resonance spectrum results are as follows: Figure 3 As shown, from Figure 3 It can be seen that 13 C NMR (101 MHz, DMSO-d 6 )δ182.21,179.91,158.90,146.25,144.32,143.90,138.21,137.30,134.78,131.12,129.59,128.73,124.80,123.11,122.42,119.90,61.72,38.11,33.52,26.12,25.43,24.92,24.01,18.68,17.52,16.01, the carbon skeleton displacement in its molecular structure was confirmed, and it can be determined to be the target product rotatable chromogenic functional molecule.

[0068] Example 2

[0069] 1. Dissolve 150.22 g (molar weight is 1 mol) of saffron aldehyde in ethanol, stir evenly at room temperature (25°C), the stirring rate is 100 rpm, the stirring time is 12 h, and a 1 mol / L saffron aldehyde solution is obtained;

[0070] 2. 318.41 g (1 mol) of coenzyme derivative Dissolve in methanol, stir evenly at room temperature (25°C) at a stirring rate of 100 rpm for 12 h to obtain a 1 mol / L coenzyme derivative solution;

[0071] 3. Disperse 101.19 g (1 mol) of triethylamine in methanol, stir thoroughly at room temperature (25°C) at a stirring rate of 100 rpm for 12 h to obtain a 1 mol / L triethylamine solution;

[0072] 4. Directly mix the coenzyme derivative solution obtained in step 2 and step 3 and the triethylamine solution, heat up directly after mixing, stir while heating, the heating rate is 10°C / h, the stirring rate is 1000rpm, after heating to 40°C, add the saffron aldehyde solution obtained in step 1 to carry out conjugated coupling reaction, wherein the addition rate of the saffron aldehyde solution is 1mL / min, the reaction time is 32h, and after the reaction is completed, a rotatable chromogenic functional molecule crude product is obtained;

[0073] 5. After the crude product of the rotatable chromogenic functional molecule obtained in step 4 is cooled to room temperature (25°C), the solvent is removed under a pressure of -0.09MPa; then, a mixed solution of dichloromethane and purified water in a volume ratio of 1:1 is used to extract the crude product of the rotatable chromogenic functional molecule from which the solvent is removed, and the organic phase is collected; then, the rotatable chromogenic functional molecule obtained by extraction is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a silica gel column filled with 200-mesh silica gel powder for purification, wherein the purification solvent is a mixed solvent of dichloromethane and methanol in a volume ratio of 1:1; then, the purified rotatable chromogenic functional molecule is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a centrifuge for centrifugal treatment once, wherein, the centrifuge The solution treated by centrifugation is a mixed solution of ethanol and purified water in a volume ratio of 1:1, the solid content of each centrifuge tube is 1 mg / mL, the centrifugal speed is 2000r / min, and the centrifugal time is 2h; then the rotatable chromogenic functional molecules obtained by centrifugation are dispersed in a mixed solution of ethanol and purified water in a volume ratio of 1:1, and are allowed to stand for crystallization at a temperature of 1°C for 1h and the solid content of the crystals is controlled to be 5mg / mL, and then the rotatable chromogenic functional molecules obtained by crystallization are subjected to vacuum filtration at a pressure of -0.09MPa, wherein the filter paper for vacuum filtration is a 1-layer slow filter paper, and the mixed solution of ethanol and purified water in a volume ratio of 1:1 is used for washing 4 times during the filtration process; finally, the rotatable chromogenic functional molecules after vacuum filtration are placed in a freeze dryer at a temperature of -50°C for drying for 1h to obtain the rotatable chromogenic functional molecules.

[0074] After testing, the yield of the rotatable chromogenic functional molecule obtained in this example is 379.9 g, and the yield is 84.3%. The nuclear magnetic resonance spectrum and mass spectrum results of the rotatable chromogenic functional molecule prepared in this example are consistent with those in Example 1.

[0075] Example 3

[0076] 1. Dissolve 751.1 g (molar weight is 5 mol) of saffron aldehyde in ethanol, stir evenly at room temperature (25°C), the stirring rate is 1000 rpm, the stirring time is 1 h, and a 5 mol / L saffron aldehyde solution is obtained;

[0077] 2. 318.41 g (1 mol) of coenzyme derivative Dissolve in propanol, stir evenly at room temperature (25°C), at a stirring rate of 1000 rpm, for 1 h, to obtain a 3 mol / L coenzyme derivative solution;

[0078] 3. Disperse 1001.2 g (10 mol) of potassium bicarbonate in acetonitrile, stir thoroughly at room temperature (25°C) at a stirring rate of 1000 rpm for 1 h to obtain a 10 mol / L potassium bicarbonate solution;

[0079] 4. Directly mix the coenzyme derivative solution obtained in step 2 and step 3 and the potassium bicarbonate solution, heat up directly after mixing, stir while heating, the heating rate is 1°C / min, the stirring rate is 2200rpm, after heating to 120°C, add the saffron aldehyde solution obtained in step 1 to carry out conjugated coupling reaction, wherein the addition rate of the saffron aldehyde solution is 10mL / min, the reaction time is 1h, and after the reaction is completed, a rotatable color-forming functional molecule crude product is obtained;

[0080] 5. After the crude product of the rotatable chromogenic functional molecule obtained in step 4 is cooled to room temperature (25°C), the solvent is removed under a pressure of -0.09MPa; then, a mixed solution of dichloromethane and purified water with a volume ratio of 15:1 is used to extract the crude product of the rotatable chromogenic functional molecule from which the solvent is removed, and the organic phase is collected; then, the rotatable chromogenic functional molecule obtained by extraction is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a silica gel chromatographic column filled with 1000 mesh silica gel powder for purification, wherein the purification solvent is a mixed solvent of dichloromethane and methanol with a volume ratio of 100:1; then, the purified rotatable chromogenic functional molecule is subjected to a pressure of -0.09MPa to remove the solvent; then, the rotatable chromogenic functional molecule from which the solvent is removed is placed in a centrifuge for centrifugal treatment 4 times, wherein each centrifugation The treated solution is a mixed solution of ethanol and purified water in a volume ratio of 15:1, the solid content of each centrifuge tube is 20 mg / mL, the centrifugal rate is 12000 r / min, and the centrifugal time is 0.1 h; then the rotatable chromogenic functional molecules obtained by centrifugation are dispersed in a mixed solution of ethanol and purified water in a volume ratio of 1:30, and crystallized at a temperature of 10°C for 24 hours and the solid content of the crystals is controlled to be 60 mg / mL, and then the rotatable chromogenic functional molecules obtained by crystallization are subjected to reduced pressure filtration at a pressure of -0.09 MPa, wherein the filter paper for reduced pressure filtration is a 3-layer slow filter paper, and the mixed solution of ethanol and purified water in a volume ratio of 1:30 is used for washing once during the filtration process; finally, the rotatable chromogenic functional molecules after reduced pressure filtration are placed in a freeze dryer at a temperature of -5°C for drying for 36 hours to obtain the rotatable chromogenic functional molecules.

[0081] After testing, the yield of the rotatable chromogenic functional molecule obtained in this example is 38.4 g, and the yield is 85.2%. The nuclear magnetic resonance spectrum and mass spectrum results of the rotatable chromogenic functional molecule prepared in this example are consistent with those in Example 1.

[0082] Application Example 1

[0083] 2.25 mg of the rotatable chromogenic functional molecule (CoenQ2-Saf) prepared in Example 1 was dissolved in malic acid to obtain a solution with a molar concentration of 5 mmol / L, and then the solution was added to three common maple syrup waters, namely maple syrup water 1, maple syrup water 2 and maple syrup water 3, and the molar concentration of the rotatable chromogenic functional molecule in each maple syrup water was controlled to be 10 μmol / L; and then the test was carried out at room temperature (25° C.), wherein the external excitation light source during the test was 350 nm, and the spectral results of the test are as follows: Figure 4 The typical values ​​of specific test data are shown in Table 1.

[0084] Table 1 Optical signal intensity and viscosity of maple syrup water

[0085] Sample Optical signal strength Viscosity Viscosity (viscometer) Maple syrup water1 3676.94 99.0cP 100.20cP Maple Syrup Water2 12374.4 498.0cP 499.33cP Maple Syrup Water3 17820.7 801.0cP 801.86cP

[0086] Depend on Figure 4 As can be seen from the data in Table 1, the viscosity of the three maple syrup waters is quite different, which leads to a large difference in the apparent light signal intensity. Specifically, the light signal intensity of maple syrup water 1 is the lowest, indicating that its consistency is thinner. Figure 4 According to the test data in Table 1 and the equation log I = 2.00 + 0.78 log η (I is the fluorescence intensity, η is the viscosity), its viscosity is 99.0 cP; the light signal of maple syrup water 2 is of medium intensity, indicating that its consistency has been improved to a certain extent, and the overall consistency is medium, Figure 4 From the test data in Table 1 and the above equation, we can see that its viscosity is 498.0 cP; the optical signal intensity of maple syrup water 3 further increases, and the overall consistency is relatively large. Figure 4 From the test data in Table 1 and the above equation, we can see that its viscosity is 801.0cP. Different consistencies indicate that its final taste and mellowness are inconsistent. High viscosity indicates that maple syrup water has a better taste and mellowness, and the overall solid content is relatively high, but its fluidity is poor; low viscosity indicates that there are more liquid components in the maple syrup water, which has better fluidity and is convenient for drinking and further dilution, but its taste and mellowness may decrease. Therefore, the size of the viscosity is closely related to the taste and mellowness of the final maple syrup water. At the same time, Figure 4 It can also be seen from the test results in Table 1 that the rotatable chromogenic functional molecular tool provided by the present invention can present apparent light signals of different intensities to maple syrup water with different micro-region viscosities, and the peak release wavelength is 510nm, which is a bright green light signal with a visualized monitoring effect.

[0087] Performance Testing:

[0088] The rotatable chromogenic functional molecular tool (CoenQ2-Saf) prepared in Example 1 was subjected to a variety of spectroscopy tests, including viscosity sensitivity test, photostability test, pH stability test, solvent polarity tolerance and detection limit test. The specific test methods and test results are as follows.

[0089] 1. Viscosity sensitivity test:

[0090] By preparing different proportions of glycerol and purified water, specifically, the proportion of glycerol and purified water is 0-99 vol%, solutions with different viscosities are obtained, namely, solutions with viscosities of 1.0 cP, 1.74 cP, 3.72 cP, 10.72 cP, 58.88 cP and 956.0 cP, the external excitation wavelength is controlled to be 350 nm, and rotatable chromogenic functional molecules with a molar concentration of 10 μmol / L diluted with malic acid are added to the above solutions, and then the viscosity sensitivity test is carried out at room temperature. The test results are as follows: Figure 5 As shown, from Figure 5 It can be seen that as the viscosity of the solution increases from 1.0 cP to 956.0 cP, the intensity of the light signal gradually increases, especially when the amount of glycerol added exceeds 50%, the intensity of the released light signal rises sharply until the volume fraction of purified water is 0%, the light signal intensity reaches the maximum value, compared with the pure water system without adding glycerol, the maximum light signal intensity increases by 339 times.

[0091] In addition, the relationship between the light signal intensity and the solution viscosity is established. After converting the light signal intensity and the solution viscosity into a logarithmic function, a straight line can be fitted, which is consistent with The specific logarithmic function value is shown in Table 2, and the specific curve is shown in Figure 6 shown.

[0092] Table 2 Logarithm of viscosity and logarithm of fluorescence intensity

[0093]

[0094] From the logarithmic results in Table 2 and Figure 6 It can be seen that the viscosity sensitivity coefficient of the rotatable chromogenic functional molecule is 0.78, and the fitting determination coefficient is 0.98, showing a high sensitivity to viscosity. It can also be seen that the rotatable chromogenic functional molecule provided by the present invention can be used as a molecular-level tool for measuring the viscosity of maple syrup water micro-areas, and the strength of the apparent light signal presented can be used to determine the size of its thinness and consistency.

[0095] 2. Light stability test:

[0096] 2.70 mg of the rotatable chromogenic functional molecule prepared in Example 1 was dissolved in malic acid to obtain a solution with a molar concentration of 6 mmol / L. During the test, the solution was diluted with malic acid to a molar concentration of 10 μmol / L, and added to low-viscosity purified water and high-viscosity glycerol, respectively. Under continuous irradiation with an external excitation light source of 350 nm, the change in light signal intensity within 60 min was tested. The test results are as follows: Figure 7 The obtained data are shown in Table 3.

[0097] Table 3 Light stability test results

[0098]

[0099]

[0100] Depend on Figure 7 From the data in Table 3, it can be seen that the photostability of the rotatable chromogenic functional molecular tool of the present invention can still maintain stable light signal release under continuous irradiation of an external excitation light source, whether in high-viscosity glycerol or low-viscosity purified water. This light signal stability indicates that it is suitable for long-term measurement of micro-area viscosity of maple syrup water, and will not be greatly affected even under long-term irradiation.

[0101] 3. pH stability test:

[0102] 0.90 mg of the rotatable chromogenic functional molecule prepared in Example 1 was dissolved in malic acid to obtain a solution with a molar concentration of 2 mmol / L. During the test, the solution was diluted with malic acid to a molar concentration of 10 μmol / L. The pH measurement range during the test was 3.0 to 12.0. Specifically, the test was performed at pH 3.0, 5.0, 6.8, 9.0 and 12.0. The test results are as follows: Figure 8 As shown, from Figure 8 It can be seen that the fluorescence intensity of the rotatable chromogenic functional molecular tool of the present invention does not change much in the pH range of 3.0 to 12.0 commonly found in maple syrup water, can show good light signal release stability, is not easily affected by pH fluctuations, and has high pH tolerance and universality.

[0103] 4. Solvent polarity tolerance test:

[0104] 3.60 mg of the rotatable chromogenic functional molecule prepared in Example 1 was dissolved in malic acid to obtain a solution with a molar concentration of 8 mmol / L. During the test, the above solution was diluted with malic acid to a molar concentration of 10 μmol / L, and the diluted solutions were added to conventional solvents of different polarities (toluene, dimethyl sulfoxide, tetrahydrofuran, dichloromethane, glycerol and ethanol), respectively. The optical signal absorption law of the rotatable chromogenic functional molecule in different polar solvent atmospheres was tested at room temperature. The test results are shown in FIG. Fig. 9 As shown, from Fig. 9 It can be seen that in solvents of different polarities, the absorbance of the rotatable chromogenic functional molecular tool of the present invention is around 0.6, and the peak of its absorption spectrum is around 350nm. The overall results show that the molecular tool is insensitive to the polarity of the solution and is suitable for regulating and monitoring the changes in the viscosity of gel water containing multiple polar components.

[0105] 5. Detection limit test:

[0106] 1.35 mg of the rotatable chromogenic functional molecule prepared in Example 1 was added to malic acid to obtain a solution with a molar concentration of 3 mmol / L. In the specific test, the above solution was diluted with malic acid to a molar concentration of 10 μmol / L, and the diluted solution was added to a mixed solution of purified water and glycerol with different viscosities to detect the lower limit of viscosity detection of the rotatable chromogenic functional molecule tool. The test results are as follows: Fig.10 As shown (I 510 is the emission fluorescence intensity value corresponding to the wavelength of 510nm). Fig.10 It can be seen that the logarithm of the viscosity value and the logarithm of the light signal release intensity of the rotatable chromogenic functional molecule of the present invention can be fitted into a straight line, the fitting coefficient of determination is 0.99, the linear equation is y=1.77+1.42x, and the detection limit of the rotatable chromogenic functional molecule is 1.13 cP through calculation, indicating that the rotatable chromogenic functional molecule of the present invention is very sensitive to the change of the viscosity of the maple syrup water micro-area, and is suitable for measuring and monitoring the viscosity of maple syrup water.

[0107] In summary, the present invention prepares a rotatable chromogenic functional molecule in one step by conjugating and coupling the coenzyme derivative and saffron aldehyde, wherein the raw material saffron aldehyde is extracted from natural saffron. The rotatable chromogenic functional molecule of the present invention has a conjugated chemical structure with alternating single and double bonds, and can present different rotation states in the solution atmosphere of different viscosities, and then convert it into a light signal and release it, so as to realize the rapid, efficient and visual detection of the viscosity (thinness) of the maple syrup water micro-area, especially for maple syrup water containing a large number of pseudoplastic macromolecular components. The traditional detection method may cause measurement errors due to shear thinning, and the molecular tool can be presented in situ through the strength of the light signal, and there will be no measurement errors, and the detection results are accurate. Moreover, the above-mentioned various test results show that the rotatable chromogenic functional molecule of the present invention has a higher sensitivity coefficient (x=0.78), better pH stability, better solvent polarity tolerance and excellent light stability. In addition, the detection limit of the rotatable chromogenic functional molecule is also low, which is suitable for the induction of weak changes in the viscosity of the maple syrup water micro-area. In addition, the preparation process of the rotatable chromogenic functional molecule is green, environmentally friendly and easy to operate, with a high yield. The raw materials are derived from natural products, which are abundant and have high added value. The dosage is in milligram level (trace amount) and the application cost is low, which is suitable for large-scale industrial production applications.

[0108] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A rotatable chromogenic functional molecule, characterized in that: The structural formula of the rotatable chromogenic functional molecule is shown in Formula I: Wherein, the Y is a coenzyme derivative group, and the coenzyme derivative group includes 2. The method for preparing a rotatable chromogenic functional molecule according to claim 1, characterized in that: The steps include: The coenzyme derivative solution, the alkaline compound solution and the safranal solution are mixed to carry out a conjugated coupling reaction to obtain a rotatable color-forming functional molecule; Wherein, the coenzyme derivative in the coenzyme derivative solution includes 3. The method for preparing a rotatable chromogenic functional molecule according to claim 2, characterized in that: The molar ratio of the coenzyme derivative in the coenzyme derivative solution, the alkaline compound in the alkaline compound solution and the saffron aldehyde in the saffron aldehyde solution is 1:1-10:1-5.

4. The method for preparing a rotatable chromogenic functional molecule according to claim 3, characterized in that: The temperature of the conjugation coupling reaction is 40-120° C., and the time is 1-32 hours.

5. The method for preparing a rotatable chromogenic functional molecule according to any one of claims 2 to 4, characterized in that: The alkaline compound in the alkaline compound solution includes one or more of sodium carbonate, cesium carbonate, aluminum hydroxide, sodium bicarbonate, potassium carbonate, potassium bicarbonate, triethylamine, trimethylamine, lithium bistrimethylsilylamide, lithium diisopropylamide, sodium bis(trimethylsilyl)amide, potassium bis(trimethylsilyl)amide, imidazole, pyridine, ammonia water and N,N,N',N'-tetramethylethylenediamine.

6. The method for preparing a rotatable chromogenic functional molecule according to claim 5, characterized in that: The solvent in the coenzyme derivative solution, the solvent in the alkaline compound solution and the solvent in the saffron aldehyde solution independently include one or more of acetonitrile, tetrahydrofuran, N,N-dimethylformamide, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, propanol, n-butanol, ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol and 1,3-butanediol.

7. The method for preparing a rotatable chromogenic functional molecule according to claim 6, characterized in that: The molar concentration of the coenzyme derivative solution is 1-3 mol / L, the molar concentration of the alkaline compound solution is 1-10 mol / L, and the molar concentration of the saffron aldehyde solution is 1-5 mol / L.

8. Use of the rotatable chromogenic functional molecule prepared by the method for preparing the rotatable chromogenic functional molecule according to any one of claims 2 to 7 in detecting the viscosity of liquid syrup fluid.

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