Elm wood shaving glue and application thereof in preparation of whitening and moisturizing products

Abstract

The invention discloses elm wood shaving glue and application thereof to preparation of whitening and moisturizing products, and relates to the field of traditional Chinese medicine, a preparation method of the elm wood shaving glue comprises the following steps: mixing elm wood shavings and deionized water, heating and extracting to obtain an extracting solution, carrying out alcohol precipitation on the extracting solution by using absolute ethyl alcohol, centrifugally collecting precipitates after alcohol precipitation, and freeze-drying. The elm wood shaving glue comprises arabinoxylan and glucose according to the mass ratio of (95-97): (3-5), and the arabinoxylan is composed of arabinose and xylose. The elm wood shaving glue has good whitening and moisturizing effects, is suitable for development of cosmetics and skin care products, and has a wide market prospect. Through a reasonable preparation method and experimental verification, the product provides a new natural component choice for the field of skin care.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine, and in particular to elm wood shavings glue and application thereof in preparing whitening and moisturizing products. Background Art

[0002] As people's demand for skin care and beauty products increases, natural ingredients with dual functions of moisturizing and whitening are gaining more and more attention. As a widely used wood, the potential value of elm extract in cosmetics needs further research and development. Summary of the invention

[0003] The purpose of the present invention is to solve at least one of the technical problems existing in the prior art, and to provide an elm wood shavings glue and its application in preparing whitening and moisturizing products.

[0004] The technical solution of the present invention is as follows: The invention discloses an application of elm wood shavings glue in preparing whitening and moisturizing products. The elm wood shavings glue comprises arabinoxylan and glucose in a mass ratio of (95-97):(3-5), and the arabinoxylan is composed of arabinose and xylose.

[0005] As a preferred embodiment of the present invention, the preparation method of the elm wood shavings glue is as follows: The elm wood shavings are mixed with deionized water, and heated for extraction to obtain an extract, and the extract is precipitated with anhydrous ethanol, and the precipitate after the alcohol precipitation is collected by centrifugation and freeze-dried.

[0006] As a preferred embodiment of the present invention, the pH value of the elm wood shavings glue is neutral.

[0007] As a preferred embodiment of the present invention, the elm wood shavings glue can reduce the accumulation level of melanin.

[0008] As a preferred embodiment of the present invention, the elm wood shavings glue can alleviate the shrinkage of the zebrafish tail.

[0009] As a preferred embodiment of the present invention, the elm wood shavings and deionized water are added in a solid-liquid ratio of 1:(8-12).

[0010] As a preferred embodiment of the present invention, the temperature of the heating extraction is 100-120°C.

[0011] As a preferred embodiment of the present invention, the extraction times are 2-4 times.

[0012] The beneficial effects of the present invention are as follows: the elm wood shavings glue of the present invention has good whitening and moisturizing effects, is suitable for the development of cosmetics and skin care products, and has broad market prospects. Through reasonable preparation methods and experimental verification, the product provides a new natural ingredient selection for the skin care field. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The effects of the samples in Example 1 on HacaT cells induced by hydrogen peroxide (H2O2); wherein, (a): elm wood shavings working solution (YM) toxicity screening on HacaT cells; (b): H2O2 toxicity screening on HacaT cells; (c): the effect of YM on the survival rate of HacaT cells induced by H2O2; (d): the effect of YM on the level of reactive oxygen species (ROS) in HacaT cells induced by H2O2; Figure 2 The effects of the samples in Example 1 on the levels of related proteins in HacaT cells induced by hydrogen peroxide; wherein, (a): the effect of YM on the expression levels of related proteins in HacaT cells induced by H2O2; (b): the expression levels of HO-1 protein in each group of HacaT cells; (c): the expression levels of Nrf2 protein in each group of HacaT cells; (d): the expression levels of Keap1 protein in each group of HacaT cells; (e): the expression levels of AQP3 protein in each group of HacaT cells; (f): the expression levels of HAS1 protein in each group of HacaT cells; (g): the expression levels of HAS2 protein in each group of HacaT cells; (h): the expression levels of HAS3 protein in each group of HacaT cells; Figure 3 The tail area of ​​zebrafish in each group; (a): Schematic diagram of the moisturizing effect of YM on the tail of zebrafish; (b): Image J statistical results of the tail area of ​​zebrafish in each group; Figure 4 The effects of the samples in the examples on B16F10 cells induced by H2O2; wherein, (a): cytotoxicity screening of elm wood shavings working solution (YM) on B16F10 cells; (b): cytotoxicity screening of H2O2 on B16F10 cells; (c): effect of YM on the survival rate of B16F10 cells induced by H2O2; (d): effect of YM on the ROS level of B16F10 cells induced by H2O2; (e): effect of YM on the tyrosinase activity of B16F10 cells induced by H2O2; (f): effect of YM on the melanin level of B16F10 cells induced by H2O2; Figure 5The figures show the effects of the samples in the examples on the expression levels of related proteins of B16F10 cells induced by hydrogen peroxide, wherein: (a): the effect of YM on the expression levels of related proteins of B16F10 cells induced by H2O2; (b): the expression levels of HO-1 protein of each group of B16F10 cells; (c): the expression levels of Keap1 protein of each group of B16F10 cells; (d): the expression levels of TYR protein of each group of B16F10 cells; (e): the expression levels of MITF protein of each group of B16F10 cells; (f): the expression levels of TRP1 protein of each group of B16F10 cells; (g): the expression levels of TRP2 protein of each group of B16F10 cells.

[0014] Figure 6 The melanin accumulation levels of zebrafish in each group, where (a): Schematic diagram of the whitening effect of YM on zebrafish; (b): Image J statistical results of melanin accumulation in zebrafish in each group; Figure 7 It is a PMP-pre-column derivatization liquid chromatogram, where 1-mannose; 2-rhamnose; 3-galacturonic acid; 4-glucose; 5-galactose; 6-xylose; 7-arabinose; Figure 8 This is the infrared spectrum of the elm wood shavings glue of Example 1; Fig. 9 This is the one-dimensional carbon spectrum of elm wood shavings glue; Fig.10 This is the one-dimensional hydrogen spectrum of elm wood shavings glue; Fig.11 This is the COSY spectrum of elm wood shavings glue; Fig.12 This is the HMBC spectrum of elm wood shavings glue; Fig.13 This is the HSQC spectrum of elm wood shavings glue; Fig.14 This is the NOESY spectrum of elm wood shavings glue. DETAILED DESCRIPTION

[0015] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used that do not specify the manufacturer are all conventional products that can be obtained commercially.

[0016] The technical solution of the present invention is described below with specific experiments.

[0017] Example 1 Preparation of elm wood shavings glue Elm wood shavings were mixed with deionized water in a solid-liquid ratio of 1:10, extracted at 100°C for 3 hours, extracted three times, impurities were filtered out with gauze, the three filtrates were combined, the alcohol concentration was adjusted to 80% with anhydrous ethanol, the precipitate after alcohol precipitation was collected by centrifugation, and freeze-dried to obtain elm wood shavings glue.

[0018] The following performance tests were performed on the elm wood shavings glue prepared in Example 1.

[0019] Test 1 Moisturizing effect test Preparation of elm shavings working solution: After confirming the solubility, stability and storage conditions of the sample to be tested, the lyophilized powder prepared in Example 1 above was prepared into a 40 mg / mL mother solution, centrifuged at 4000 rpm for 5 min, the insoluble precipitate was discarded, and the supernatant was diluted into a series of working solutions of different concentrations.

[0020] Preparation of standard dilution water for zebrafish experiment: weigh the calculated reagents according to the mass ratio: sodium chloride 8g / L, potassium chloride 0.2g / L, calcium chloride 0.24g / L, magnesium sulfate 0.2g / L, sodium bicarbonate 1.2g / L, add to deionized water, stir until the reagents are dissolved, and adjust the pH value of the solution to 7.2-7.4. Filter with a 0.22µm filter, store in a sterile bottle, and store in a 4℃ refrigerator.

[0021] (1) Verification of cell moisturizing efficacy A. Modeling drug: hydrogen peroxide (H2O2). A preliminary experiment should be conducted to confirm that the survival rate of HacaT cells in the model group is significantly lower than that in the control group before it can be used as a modeling drug. The specific modeling method is as follows: HaCaT cells are cultured at a rate of 1×10 4 100 μL of MTT solution were added to each well and incubated in a 37°C incubator for 3 h. The culture medium in all wells was removed and 200 μL of dimethyl sulfoxide (DMSO) was added to each well in turn. The culture plate was wrapped with tin foil and placed on a shaker and incubated for 3 h in the dark.

[0022] B. Therapeutic drugs: elm wood shavings glue working solution. A preliminary experiment should be done first to determine the maximum detection concentration of the sample to be tested, which provides a reference for the concentration setting of the formal experiment. The stock solution of the sample to be tested is diluted with standard dilution water in a geometric series of ≤2.2 times to different concentrations, which are 3.125, 6.25, 12.5, 25, 50, 100, 200, 400, 800, and 1600 μg / mL.

[0023] C. Experimental Grouping: Control group: HacaT cells and PBS.

[0024] Model group: HacaT cells, H2O2, and PBS. The working concentration of H2O2 was 500µM.

[0025] Sample group: HacaT cells, H2O2, and elm wood shavings glue working solution.

[0026] D. Formal experiment: HacaT cells were used to construct the model, and 500µM H2O2 was used to model the model. A control group, a model group, and a sample group were set up. HaCaT cells were cultured at a concentration of 1×10 4 The cells were inoculated into 96-well culture plates at 100 μg / well. When the cells grew to 80%-90% of the culture dish, PBS was added to the control group and the model group, and elm wood shavings glue working solution (6.25, 12.5, 25, 50, 100 μg / mL) was added to the sample group for incubation for 1 h. Then H2O2 (500 μM) was added to the model group and the sample group, and PBS was added to the control group. After incubation at 37°C for 24 h, the survival rate of HacaT cells in each group was calculated by MTT method; the reactive oxygen species (ROS) level of HacaT cells in each group was detected by DCFHDA method; the expression levels of Keap1, Nrf2, HO-1, HAS1, HAS2, HAS3, and AQP3 in HacaT cells in each group were detected by Western Blot method.

[0027] E. Statistical Analysis Use SPSS software or any other software to perform statistical analysis on the test data, and use the model group as the standard to compare the differences in cell survival rate, ROS, and protein expression levels between the model group and each experimental group. If the test data is normally distributed, use t The test method is used for statistical analysis; if it is a non-normal distribution, the rank sum test method is used for statistical analysis, and the statistical method uses a two-tailed test. p <0.05 was considered to be significantly different.

[0028] F. Experimental Results Results Figure 1-Figure 2 As can be seen from the figure, compared with the control group, the cell survival rate of the H2O2-induced model group was significantly reduced, and the cell survival rate was significantly increased and the ROS level was significantly reduced after intervention with elm wood shavings glue; further detection of related proteins found that elm wood shavings glue can increase the expression levels of Keap1, Nrf2, and AQP3 proteins in HacaT cells induced by H2O2, and reduce the expression levels of HO-1, HAS1, HAS2, and HAS3 proteins.

[0029] (2) Verification of the moisturizing effect of zebrafish A. Modeling drug: sodium chloride. A preliminary experiment should be done to determine that the tail area of ​​zebrafish in the model group is reduced and significantly different from that in the control group before it can be used as a modeling drug. The specific modeling process is as follows: wild AB strain zebrafish, select normal zebrafish embryos under a stereomicroscope, and place them in a 1 mg / mL Pronase® protease solution for about 3 minutes. When the fertilized eggs develop to 2 dpf, remove the outer egg membrane of the zebrafish embryo; add sodium chloride solution and culture in a constant temperature incubator at 28°C for 24 hours.

[0030] B. Positive drug: Sodium hyaluronate. A preliminary experiment should be conducted to confirm that the tail area of ​​zebrafish in the positive group is increased and significantly different from that in the model group before it can be used as a positive drug.

[0031] C. Therapeutic drugs: elm wood shavings glue working solution. A preliminary experiment should be done first to determine the maximum detection concentration of the sample to be tested, which provides a reference for the concentration setting of the formal experiment.

[0032] D. Experimental Grouping: Control group: contained zebrafish and standard dilution water.

[0033] Model group: Contains sodium chloride, zebrafish and standard dilution water. The working concentration of sodium chloride is 1% (g / mL).

[0034] Positive group: Contains sodium hyaluronate, zebrafish and standard dilution water. The working concentration of sodium hyaluronate is 0.1% (g / mL).

[0035] Sample group: Contains elm wood shavings glue working solution, zebrafish and standard dilution water.

[0036] E. Formal experiment: 2dpf AB zebrafish were used to construct the model, the membrane of zebrafish embryos was removed, and 1% sodium chloride was used to model. The control group, model group, positive group, and sample group were set up. Wild AB zebrafish were selected, normal zebrafish embryos were placed in 1 mg / mL Pronase® protease solution for about 3 min under a stereomicroscope. When the fertilized eggs developed to 2 dpf, the outer egg membrane of the zebrafish embryo was removed. The treated zebrafish were randomly divided into a control group, a model group (1% sodium chloride), a drug group (elm wood shavings glue working solution 2.5, 5mg / mL), and a positive group (0.1% sodium hyaluronate), with 10 tails in each group. Except for the control group (zebrafish breeding water), sodium chloride solution was added to all other groups and cultured in a constant temperature incubator at 28℃ for 24h. 8-10 zebrafish were randomly selected from each group, observed and photographed under a microscope for analysis, and the changes in the tail area of ​​zebrafish were observed and recorded to evaluate the moisturizing effect of elm wood shavings glue.

[0037] F. Statistical Analysis Use SPSS software or any other software to perform statistical analysis on the test data, and use the model group as the standard to compare the difference in zebrafish tail area between the model group and each experimental group. t The test method is used for statistical analysis; if it is a non-normal distribution, the rank sum test method is used for statistical analysis, and the statistical method uses a two-tailed test. p <0.05 was considered to be significantly different.

[0038]

[0039] G. Experimental Results Results Figure 3 Compared with the control group, the tail area of ​​zebrafish in the model group was significantly reduced and severely wrinkled; after intervention with elm wood shavings glue, the wrinkling of the zebrafish tail was alleviated.

[0040] 2. Whitening effect verification (1) Verification of cell whitening efficacy A. Modeling drugs: H2O2 should be used as a modeling drug after a preliminary experiment to confirm that the survival rate of B16F10 cells in the model group is significantly lower than that in the control group. The specific modeling process is as follows: B16F10 cells were cultured at 1×10 4 100 μL of MTT solution were added to each well of the 96-well culture plate. After culturing in the incubator for 24 h, different concentrations of H2O2 (31.25, 62.5, 125, 250, 500, 1000 μM) were added to each well to treat the cells. After incubation at 37 °C for 24 h, 50 μL of 2 mg / mL MTT solution was added to each well and incubated in a 37 °C incubator for 3 h. The culture medium in all wells was removed, and 200 μL of dimethyl sulfoxide (DMSO) was added to each well in turn. The culture plate was wrapped with tin foil and placed on a shaker, incubated for 3 h in the dark.

[0041] B. Therapeutic drugs: elm wood shavings glue working solution. A preliminary experiment should be done first to determine the maximum detection concentration of the sample to be tested, which provides a reference for the concentration setting of the formal experiment. The stock solution of the sample to be tested is diluted with standard dilution water in a geometric series of ≤2.2 times to different concentrations, which are 3.125, 6.25, 12.5, 25, 50, 100, 200, 400, 800, and 1600 μg / mL.

[0042] C. Experimental Grouping: Control group: B16F10 cells and PBS.

[0043] Model group: B16F10 cells, H2O2, and PBS. The working concentration of H2O2 was 250μM.

[0044] Sample group: B16F10 cells and H2O2, elm wood shavings glue working solution.

[0045] D. Formal experiment: B16F10 cells were used to construct the model, H2O2 (125 μM) was used to model, and a control group, a model group, and a sample group were set up. B16F10 cells were cultured at a rate of 1×10 4 The cells were inoculated into 96-well culture plates at 100 μg / well. When the cells grew to 80%-90% of the culture dish, PBS was added to the control group and the model group, and elm wood shavings glue working solution (6.25, 12.5, 25, 50, 100 μg / mL) was added to the sample group for incubation for 1 h. Then H2O2 (125 μM) was added to the model group and the sample group, and PBS was added to the control group. After incubation at 37°C for 24 h, the survival rate of B16F10 cells in each group was calculated by MTT method; the level of reactive oxygen was detected by DCFHDA method; the tyrosinase activity and melanin accumulation level were detected; the expression levels of Keap1, HO-1, MITF, TYR, TRP1, and TRP2 in B16F10 cells in each group were detected by Western Blot method.

[0046] E. Statistical Analysis Use SPSS software or any other software to perform statistical analysis on the test data, and use the model group as the standard to compare the differences in cell survival rate, reactive oxygen species, tyrosinase activity, melanin accumulation, and protein expression levels between the model group and each experimental group. If the test data is normally distributed, use t The test method is used for statistical analysis; if it is a non-normal distribution, the rank sum test method is used for statistical analysis, and the statistical method uses a two-tailed test. p <0.05 was considered to be significantly different.

[0047] F. Experimental Results Results Figure 4-Figure 5 Compared with the control group, the cell survival rate of the hydrogen peroxide-induced model group was significantly decreased. After intervention with elm wood shavings glue, the cell survival rate was significantly increased, the reactive oxygen level was significantly decreased, the tyrosinase activity was significantly decreased, and the melanin accumulation water was significantly decreased. Further detection of related proteins showed that elm wood shavings glue could increase the expression level of Keap1 protein in B16F10 cells induced by hydrogen peroxide, and reduce the expression levels of HO-1, MITF, TYR, TRP1, and TRP2 proteins.

[0048] (2) Verification of zebrafish whitening efficacy A. Modeling drug: α-melanocyte stimulating hormone (α-MSH). A preliminary experiment should be done first to determine that the melanin content of zebrafish in the model group is increased and significantly different from that in the control group before it can be used as a modeling drug. The specific modeling process is as follows: 7-9 hpf zebrafish embryos are randomly divided into a control group and a model group, and zebrafish are given α-MSH induction treatment every day according to the group until 72 hpf. The 72 hpf zebrafish are placed under a microscope and photographed to observe the distribution of melanin in the zebrafish.

[0049] B. Positive drug: α-arbutin. A preliminary experiment should be conducted to confirm that the melanin content of zebrafish in the positive group is significantly lower than that in the model group before it can be used as a modeling drug.

[0050] C. Therapeutic drugs: elm wood shavings glue working solution. A preliminary experiment should be done first to determine the maximum detection concentration of the sample to be tested, which provides a reference for the concentration setting of the formal experiment.

[0051] D. Experimental Grouping: Control group: contained zebrafish and standard dilution water.

[0052] Model group: Contains α-MSH, zebrafish and standard dilution water. The working concentration of α-MSH is 3 mg / mL.

[0053] Positive group: Contains α-arbutin, zebrafish and standard dilution water. The working concentration of α-arbutin is 3 mg / mL.

[0054] Sample group: Contains elm wood shavings glue working solution, zebrafish and standard dilution water.

[0055] E. Formal experiment: AB zebrafish at 7-9 hpf were induced with 1.5μM α-MSH and treated with drugs according to the groups. The culture medium was renewed every 24 hours until 72 hpf, when photos were taken and records were taken. A control group, model group, positive group, and sample group were set up to observe and record the melanin content of zebrafish and evaluate the whitening effect of elm wood shavings glue.

[0056] F. Statistical Analysis SPSS software or any other software was used to perform statistical analysis on the test data, and the model group was used as the standard to compare the difference in the amount of melanin produced in the zebrafish skin between the model group and each experimental group. If the test data was normally distributed, the t-test method was used for statistical analysis; if it was non-normally distributed, the rank sum test method was used for statistical analysis. The statistical methods all used two-tailed tests, and p<0.05 was considered to be significantly different.

[0057]

[0058] G. Experimental Results Results Figure 6 Compared with the control group, the melanin accumulation level of zebrafish in the model group was significantly increased; after intervention with elm wood shavings glue, the melanin accumulation level of zebrafish was significantly reduced.

[0059] Test 2: Structural test (1) Chemical composition of elm wood shavings glue: The chemical composition (water content, ash content, protein, fat) of the elm shavings glue prepared in Example 1 was determined according to the pharmacopoeia method; the total sugar content of the elm shavings glue was determined by the phenol-sulfuric acid method; and the monosaccharide composition was determined by ion chromatography. The results are shown in Table 1. The total sugar content of the elm shavings glue was 96.30%, indicating that carbohydrates were the main component of the elm shavings glue.

[0060] Table 1 Chemical composition of elm wood shavings glue

[0061] (2) Molecular weight distribution Using the molecular weight of the reference substance as the ordinate and the corresponding retention time as the abscissa, the GPC software was used to draw a standard curve and calculate the regression equation: y =16.663281-0.909183 x + 0.016227 x 2 - 0.000061 x 3 , and the molecular weights of elm wood shavings are shown in Table 2.

[0062] Table 2 Molecular weight distribution of elm wood shavings glue

[0063] The monosaccharide composition analysis is as follows: The results of the determination of monosaccharide composition in elm wood shavings rubber samples are shown in Figure 7 , the peak area percentage of each monosaccharide component was calculated by normalization method (see Table 3). The results showed that the elm wood shavings gum sample was mainly composed of arabinose and xylose, and the total molar proportion of the two monosaccharides exceeded 96%, so it was defined as arabinoxylan; the monosaccharide with a smaller content was glucose, accounting for about 3%. It is worth noting that no uronic acid was found in the composition, confirming that the polysaccharide is neutral.

[0064] Table 3 Test results of monosaccharide composition of polysaccharides

[0065] (3) Infrared analysis The infrared spectrum results of elm wood shavings glue are shown in Figure 8 . The infrared spectrum data were analyzed by using Omnic software and combined with literature. Generally speaking, 3700-3100 cm-1 (3391.34 cm -1 The strong broad peak that appears is the stretching vibration peak of OH, 3000~2800 cm -1 (2934.35 cm -1 The strong shoulder peak of ) is derived from the CH asymmetric stretching vibration peak in the sugar ring. These two are typical characteristic peaks of polysaccharides. -1 (1641.20 cm -1 ), indicating the presence of the asymmetric stretching vibration peak of the carbonyl C=O. At the same time, combined with the 1465-1360 cm -1 (1333.46 cm -1 ) shows that the polysaccharide contains uronic acid. 1040.99 cm -1 The absorption peak at 901.40 cm -1 The strong absorption peak at 840.93 cm -1 The sharp peak at indicates the possible existence of α-glycosidic bond. The infrared spectrum of elm wood shavings glue shows that elm wood shavings glue is a polysaccharide containing carboxyl, β-glycosidic bond, pyran ring structure, and possibly α-glycosidic bond.

[0066] (4) Methylation analysis The monosaccharide composition and glycosidic bond composition were further verified by methylation experiments. From the results in Table 4, we can see that the main residual sugar components are arabinose and xylose, and the total ratio is close to the monosaccharide composition, thus verifying the results of the monosaccharide composition; the residual sugar composition of arabinose is mainly residual sugar 1,3-Ara f and t-Ara f It is speculated that the main branched end sugar of the polysaccharide is furanose arabinose, and there is also about 8% pyranose arabinose end sugar, which is speculated to be the connecting component with xylose; in the other main component monosaccharide xylose, the main residual sugar is pyranose 2,3,4-Xyl p , indicating that xylose constitutes the main chain part of the polysaccharide and there are multi-branched residual sugars.

[0067] Table 4 Methylation test results of polysaccharides

[0068] (5) NMR analysis The structure of the polysaccharide was confirmed and represented by multiple one-dimensional and two-dimensional NMR spectra. 1In the anomeric region of H NMR (4.3-5.8 ppm), multiple groups of anomeric hydrogen H1 signals can be found, and the main peak positions are identified as 5.16, 5.03, 4.94, 4.89, 4.34 ppm, etc., confirming that the polysaccharide is composed of multiple residual sugars. In the 3.0-4.3 ppm range of the hydrogen spectrum, there are alkyl proton signals in the residual sugar structure except for the anomeric hydrogen. The signals in this region are numerous and mixed, which conforms to the general characteristics of the hydrogen spectrum of polysaccharides. No obvious methyl signals were found in the high-field region of the hydrogen spectrum, which corresponds to the result of the absence of rhamnose and other methylated residual sugars in the methylation.

[0069] Solid-state NMR carbon spectrum of the polysaccharide 13 In C NMR, no uronic acid carbonyl signal was found in the range of 170-210 ppm, which echoed the results of no acidic residual sugar in the monosaccharide composition and methylation results. The anomeric region of the carbon spectrum (90-110 ppm) mainly identified three groups of main anomeric carbon C1 signals, which were 108.42, 101.68, and 98.66 ppm, respectively, confirming that the polysaccharide is composed of at least three or more residual sugars; in the wide range of 58-90 ppm, there are multiple carbon peak signals of polysaccharide residual sugars except anomeric carbon, among which multiple signals near 63 ppm are attributed to the methylene -CH2- outside the five-membered pentose residual sugar ring or the C5 secondary carbon of the six-membered pentose residual sugar, and multiple signals near 80-85 ppm are attributed to multiple carbon positions of the five-membered ring residual sugar.

[0070] The structure of the polysaccharide was further analyzed by two-dimensional NMR spectrum. In the HSQC two-dimensional NMR spectrum reflecting the connection between hydrogen and carbon, multiple groups of H1 / C1 related signals in the anomeric region were identified, which were 5.16 / 108.39, 5.06 / 101.08, 4.97 / 99.41, 4.93 / 98.49 ppm, etc. The anomeric related signals at other positions were weak, which confirmed that they were residues with low content. According to the integral intensity of the anomeric hydrogen of the hydrogen spectrum and the residual sugar ratio of the methylation result, combined with the peak signals of previous research literature, the comprehensive correspondence of the main anomeric positions and residues was judged to be 5.16 / 108.39 for α-t-Ara f -(1→H1 / C1 signal, 5.06 / 101.08 is α-1,3-Ara f -(1→H1 / C1 signal, 4.97 / 99.41 for α-t-Xyl p -(1→ and α-t-Ara p -(1→H1 / C1 overlap signal, 4.93 / 98.49 for β-1,2,3,4-Xyl p -(1→H1 / C1 signal.

[0071] By combining multiple 2D NMR spectra, the positions and structural correspondence information of the carbon-hydrogen signals of various residual sugars in the arabinoxylan structure were assigned in turn. For example, the anomeric proton signal of 5.16 ppm was determined to be attributed to the residual sugar α-t-Ara f -(1→, the anomeric H1 of 4.27 ppm was found to be strongly correlated with it by identifying the COSY two-dimensional spectrum related to hydrogen and hydrogen and combining it with the NOESY spectrum, confirming that the positions of H1 and H2 were 5.16 and 4.27 ppm; combined with the HSQC spectrum, the signal of H2 / C2 was confirmed to be 4.27 / 79.12; according to the information of COSY and NOESY spectra, the signal related to H2 and H3 was found to be 4.27 / 3.87, confirming that the hydrogen spectrum position of C3 was at 3.87 ppm, and combined with HSQC to find 3.87 / 75.68, confirming that the position of C3 was 75.68 ppm, and the position information of H4 / C4 and H5 / C5 was determined to be 4.10 / 75.73 and 3.61 / 62.76 by the same method. According to this method, the positions of the other four main residual sugars were found, and the signals were assigned accordingly, forming the following Table 5: Table 5 Main residues of arabinoxylan

[0072] The glycosidic bonds in the structure of the arabinoxylan were determined by HMBC and NOESY 2D spectra. f -(1→ In the residual sugar, the anomeric hydrogen related signal was found in the spectrum: 5.18 / 77.35. According to the above residual sugar position assignment, it was confirmed that the anomeric hydrogen H1 of the residual sugar was related to β-1,2,3,4-Xyl p -(1→C3 correlation of residual sugars confirms the presence of α-t-Ara f -(1→3)-β-1,2,3,4-Xyl p -(1→ connected branched chain structure fragment, the glycosidic bond constitutes a part of the branch chain, NOESY related signal 5.16 / 3.90 confirms the glycosidic bond; NOESY signal 5.04 / 3.61 speculates that there is α-1,3-Ara in the structure constituting the main chain f -(1→4)-β-1,2,3,4-Xyl p -(1→ structural fragment; at the same time for α-t-Ara f Residual sugar, found NOE signal of 5.16 / 3.92, inferred the presence of α-t-Ara f -(1→3)-α-1,3-Ara f -(1→ structural fragment, confirming the presence of α-1,3-Ara fResidual sugar, another thing worth noting is the residual sugar α-1,3-Ara f -(1→ NOE signal 5.06 / 3.73 confirmed the possible existence of α-t-Ara f -(1→3)-α-1,3-Ara f -(1→2)-β-1,2,3,4-Xyl p -(1→fragment; NOE signals 4.95 / 3.73 and 4.96 / 3.90 confirmed the presence of α-t-Ara p -(1→2)-β-1,2,3,4-Xyl p -(1→ and α-t-Xyl p -(1→3)-β-1,2,3,4-Xyl p -(1→ glycosidic bonds, these two glycosidic bonds constitute short branches in the main chain. In terms of the self-correlation of residual sugars, NOE signals 5.06 / 3.92 and 4.93 / 3.61 confirm that α-1,3-Ara f -(1→ and β-1,2,3,4-Xyl p -(1→) has glycosidic bonds with its own residual sugars 1→3 and 1→4.

[0073] Based on the above structural analysis, it is speculated that the arabinoxylan has a high degree of branching, and its main structure is shown in the following formula. Figure 9-Figure 14 : Fig. 9 The response is a one-dimensional carbon spectrum. Fig.10 The response is a one-dimensional hydrogen spectrum. Fig.11 The COSY spectrum reflects the coupling relationship between HH; Fig.12 The HMBC spectrum reflects the coupling relationship between long-range HCs; Fig.13 The HSQC spectrum reflects the relationship between directly connected CH; Fig.14 The NOESY spectrum reflects the spatial coupling effect between the various hydrogens.

[0074] .

[0075] In summary, the elm wood shavings glue obtained by the present invention has good whitening and moisturizing effects and is suitable for the development of cosmetics and skin care products. Through reasonable preparation methods and experimental verification, this product provides a new natural ingredient selection for the skin care field.

[0076] The above-mentioned embodiments only express the preferred implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for ordinary technicians in this field, various other corresponding changes and deformations can be made according to the technical solutions and concepts described above, and all these changes and deformations should fall within the scope of protection of the claims of the present invention.

Claims

1. An application of elm wood shavings glue in preparing whitening and moisturizing products, characterized in that: The elm wood shavings glue comprises arabinoxylan and glucose in a mass ratio of (95-97):(3-5), wherein the arabinoxylan is composed of arabinose and xylose.

2. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 1, characterized in that: The preparation method of the elm wood shavings glue is as follows: The elm wood shavings are mixed with deionized water, and heated for extraction to obtain an extract, and the extract is precipitated with anhydrous ethanol, and the precipitate after the alcohol precipitation is collected by centrifugation and freeze-dried.

3. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 1, characterized in that: The pH value of the elm wood shavings glue is neutral.

4. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 1, characterized in that: The elm wood shavings glue can reduce the accumulation level of melanin.

5. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 1, characterized in that: The elm wood shavings glue can alleviate the shrinkage of the zebrafish tail.

6. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 2, characterized in that: The elm wood shavings and deionized water are added in a solid-liquid ratio of 1:(8-12).

7. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 2, characterized in that: The temperature of the heating extraction is 100-120°C.

8. The use of elm wood shavings glue in the preparation of whitening and moisturizing products according to claim 2, characterized in that: The extraction times are 2-4 times.

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