Anti-inflammatory soothing turmeric stock solution as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of plant processing and extraction, and particularly relates to an anti-inflammatory soothing turmeric stock solution as well as a preparation method and application thereof. The invention provides a preparation method of an anti-inflammatory soothing turmeric stock solution. The preparation method comprises the following steps: S1, pretreating turmeric to obtain turmeric to be squeezed; s2, squeezing the rhizoma curcumae longae to be squeezed by using a squeezer to obtain a rhizoma curcumae longae liquid A; s3, adding a preservative into the turmeric liquid A, and then adjusting the pH value to obtain turmeric liquid B; s4, centrifuging the turmeric liquid B to obtain supernatant; and S5, adding a suspending agent into the supernatant to obtain the turmeric stock solution. The invention also provides the turmeric stock solution obtained by the preparation method. A squeezing method is applied to preparation of the turmeric extract, and a 100% turmeric stock solution is obtained; compared with a traditional water extraction method, more active matters insoluble in water are reserved, so that the content of total polyphenols and total flavonoids in the turmeric stock solution is higher, and the turmeric stock solution has the effect of reducing the level of inflammatory factors.

Description

Technical Field

[0001] The invention belongs to the technical field of plant processing and extraction, and particularly relates to an anti-inflammatory and soothing turmeric stock solution and a preparation method and application thereof. Background Art

[0002] Turmeric (Curcuma longa L.) is a perennial herb of the genus Curcuma in the Zingiberaceae family. It is a traditional Chinese medicine with anti-cancer, antioxidant, cardiovascular disease prevention, and antibacterial effects. The main components of turmeric are curcumin and volatile oil compounds, as well as flavonoids, oils, sugars, sterols, peptides, metal elements, etc. Due to the presence of these natural ingredients, turmeric can have anti-aging and anti-wrinkle effects in cosmetics. Inflammation is related to hair loss such as male pattern baldness and female pattern baldness. Relieving scalp or hair follicle inflammation may be one of the strategies to alleviate hair loss.

[0003] The traditional method of extracting active ingredients from turmeric is mainly the solvent method, which uses organic solvents such as water, ethanol or acetone for extraction. However, the traditional extraction method requires high temperature extraction, which pollutes the environment to a certain extent; and it is difficult to obtain 100% turmeric stock solution using the traditional extraction method. Some effective ingredients that are difficult to dissolve in water (such as curcumin, flavonoid active substances, etc.) cannot be retained in the solution obtained by the water extraction method. Summary of the invention

[0004] In order to solve the above technical problems, the present invention adopts the following technical solutions: The present invention provides a method for preparing an anti-inflammatory and soothing turmeric stock solution, comprising the following steps: S1. Pre-treating turmeric to obtain turmeric to be squeezed; S2, squeezing the turmeric to be squeezed with a squeezer to obtain turmeric liquid A; S3, adding a preservative to the turmeric solution A and adjusting the pH, collecting and obtaining turmeric solution B; S4, centrifuging the turmeric solution B to obtain a supernatant; S5. Add a suspending agent to the upper layer liquid to obtain an anti-inflammatory and soothing turmeric stock solution.

[0005] Furthermore, the suspending agent includes one or both of xanthan gum and hydroxypropyl methylcellulose.

[0006] Furthermore, the mass ratio of the suspending agent to the turmeric solution A is 0.1% to 1.0%; and the solid content of the anti-inflammatory and soothing turmeric stock solution is 2.7% to 4.0%.

[0007] Furthermore, in the anti-inflammatory and soothing turmeric stock solution, the total polyphenol content includes 2.4-3.0 mg / ml, and the total flavonoid content includes 1.6-2.1 mg / ml.

[0008] Furthermore, the press in step S2 uses a pressing plate as a pressing component; the pressing pressure in step S2 includes 10 to 15 MPa; the single pressing time in step S2 includes 10 to 30 minutes; the number of pressings in step S2 includes 3 to 7 times; the centrifugal speed in step S4 includes 1000 to 10000 rmp / min, and the time includes 30 minutes.

[0009] Furthermore, the pretreatment in step S1 includes washing, drying, disinfecting, and cutting in sequence; the preservative in step S3 includes a mixture of phenoxyethanol and sodium benzoate, or a mixture of phenoxyethanol, benzoic acid, and dehydroacetic acid; in step S3, the pH is adjusted to 5.5; and step S5 includes: adding the suspending agent to the upper layer liquid, mixing evenly, and obtaining the anti-inflammatory and soothing turmeric stock solution.

[0010] Furthermore, the slicing in step S1 includes dicing; the mass ratio of the preservative in step S3 to the turmeric liquid A in step S2 includes 1% to 1.1%; compared with the mass of the turmeric liquid A, the preservative includes a mixture of 0.8% by mass of phenoxyethanol and 0.2% by mass of sodium benzoate, or a mixture of 0.9% by mass of phenoxyethanol, 0.12% by mass of benzoic acid and 0.08% by mass of dehydroacetic acid; citric acid is used to adjust the pH in step S3; and the mixing method in step S5 includes stirring.

[0011] The present invention also provides an anti-inflammatory and soothing turmeric stock solution obtained by the preparation method of the anti-inflammatory and soothing turmeric stock solution.

[0012] The present invention also provides an application of the anti-inflammatory and soothing turmeric stock solution, which is used for preparing a preparation with anti-inflammatory and soothing effects.

[0013] Furthermore, the anti-inflammatory effect includes inhibiting the expression of inflammatory factor IL-1α by keratinocytes under external stimulation.

[0014] Furthermore, the turmeric used in the present invention is fresh turmeric.

[0015] Furthermore, the temperature of the preparation method of the present invention is room temperature.

[0016] Furthermore, the solid content is calculated by mass.

[0017] The solid content determination method of the anti-inflammatory and soothing turmeric stock solution of the present invention is as follows: take out a weighing bottle from a dryer, weigh and record it as m1, add an appropriate amount of sample (turmeric stock solution), weigh and record it as m2, put it into an oven, bake it at 105° C. for 3 hours, take it out, put it into a dryer to cool, weigh and record it as m3, solid content = (m3-m1) / (m2-m1)*100%, and the solid content is calculated according to the solid content formula.

[0018] The preparation method of the invention adopts a pressing method and does not require solvent extraction to obtain 100% turmeric stock solution.

[0019] The mixed solution obtained after squeezing of the present invention has residues and the like remaining in it. Although centrifugation can remove starch, protein and residues, the supernatant after centrifugation still has precipitation after long-term storage. The turmeric stock solution of the present invention adds suspending agents such as xanthan gum to make the system more uniform and stable, and there is no precipitation after storage. The use of suspending agents stabilizes more water-insoluble active substances. Therefore, the anti-inflammatory and soothing turmeric stock solution obtained by the method of the present invention has a higher content of active ingredients than the solution obtained by water extraction and conventional squeezing methods, and accordingly, the anti-inflammatory effect is also better.

[0020] Keratinocytes are one of the immune cells of the skin / scalp. After being stimulated by external factors, keratinocytes can release IL-1α, causing biochemical reactions in other immune cells of the skin, inducing symptoms such as itching, inflammation, and redness. The anti-inflammatory and soothing turmeric stock solution of the present invention can inhibit the expression of keratinocyte pro-inflammatory factor IL-1α at a specific concentration, and has anti-inflammatory and soothing effects.

[0021] Studies have shown that IL-1α levels are related to hair cycles and are negative regulators of hair growth. They may be involved in the transition from growth phase to catagen phase, and also in the induction of growth phase. The IL-1α level of hair follicles in the growth phase is relatively lower. Therefore, the anti-inflammatory and soothing turmeric stock solution of the present invention is mild and non-irritating, has anti-inflammatory and soothing effects, and is suitable for the development of anti-hair loss and soothing scalp care products.

[0022] Compared with the prior art, the implementation of the present invention has the following beneficial effects: 1. The present invention applies the pressing method to the preparation of turmeric extract to obtain 100% anti-inflammatory and soothing turmeric stock solution, filling the technical gap in this field.

[0023] 2. Compared with the traditional water extraction method or the conventional pressing method, the method of the present invention retains and stabilizes more water-insoluble active substances, so that the total polyphenol and total flavonoid content in the turmeric stock solution of the present invention is higher.

[0024] 3. The method of the present invention does not require high-temperature preparation and is safer and more environmentally friendly than other methods.

[0025] 4. The anti-inflammatory and soothing turmeric stock solution of the present invention has the effect of inhibiting the expression of the inflammatory factor IL-1α of keratinocytes under external stimulation and reducing the level of inflammatory factors.

[0026] 5. The anti-inflammatory and soothing turmeric stock solution of the present invention is mild, non-irritating, and free of pesticide residues. It can be applied to the formulation of scalp care products, especially anti-hair loss products, and has the effect of preventing consumers' hair from falling out. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The endpoint is that 10% of the anti-inflammatory and soothing turmeric stock solution of the present invention acts on the CAM membrane for 5 minutes; Figure 2 A partial picture of the press used in the present invention (pressing plate); Figure 3 The cytotoxicity test results of the anti-inflammatory and soothing turmeric stock solution of the present invention; Figure 4 The anti-inflammatory and soothing test experiment grouping of the anti-inflammatory and soothing turmeric stock solution of the present invention; Figure 5 The anti-inflammatory and soothing test IL-1α comparison results of the anti-inflammatory and soothing turmeric stock solution of the present invention. DETAILED DESCRIPTION

[0028] To make the purpose, technical scheme and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific implementation methods and drawings. Unless otherwise specified, the reagents and materials used in the following examples are conventional commercial products.

[0029] The relevant information of the press used in the embodiment of the present invention is as follows: Experimental press (5L, 20T tonnage), customized by Zhucheng Hengbin Machinery Co., Ltd.; the press plate is used as the main pressing component to press the plant, the press plate picture is shown in Figure 2 .

[0030] Example 1 (1) Wash, dry and disinfect the turmeric.

[0031] (2) Weigh 610 g of detoxified turmeric, cut it into cubes, put it into a pressing bag, and place it into the press barrel.

[0032] (3) Set the pressing pressure range to 13-15 MPa and continue pressing for 20 min. Take the residue for the next pressing. Repeat the pressing for 5 times and collect the liquid. A total of 212 g of turmeric liquid A was obtained, with a pressing rate of 34.8%.

[0033] (4) Add 1% preservative by weight (0.8% phenoxyethanol, 0.2% sodium benzoate).

[0034] (5) Add appropriate amount of citric acid to adjust the pH to 5.5.

[0035] (6) Centrifuge (3000 rpm, 30 min), add 0.3% by weight of suspending agent (0.2% xanthan gum + 0.1% hydroxypropyl methylcellulose), stir well to obtain turmeric stock solution, and measure the solid content of the turmeric stock solution to be 3.93%.

[0036] Example 2 (1) Wash, dry and disinfect the turmeric.

[0037] (2) Weigh 613 g of detoxified turmeric, cut it into cubes, put it into a pressing bag, and place it into the press barrel.

[0038] (3) Set the pressing pressure range to 13-15 MPa and continue pressing for 20 min. Take the residue for the next pressing. Repeat the pressing for 7 times and collect the liquid. A total of 344 g of turmeric liquid A was obtained, with a pressing rate of 56.1%.

[0039] (4) Add 1% preservative by weight (0.8% phenoxyethanol, 0.2% sodium benzoate).

[0040] (5) Add appropriate amount of citric acid to adjust the pH to 5.5.

[0041] (6) Centrifuge (3000 rpm, 30 min), add 0.1% by weight of suspending agent (xanthan gum), stir thoroughly to obtain turmeric stock solution, and measure the solid content of the turmeric stock solution to be 3.15%.

[0042] Example 3 (1) Wash, dry and disinfect the turmeric.

[0043] (2) Weigh 807.8 g of detoxified turmeric, cut it into cubes, put it into a pressing bag, and place it into the press barrel.

[0044] (3) Set the pressing pressure range to 10-15 MPa and continue pressing for 20 min. Take the residue for the next pressing. Repeat the pressing for 5 times and collect the liquid. A total of 540 g of turmeric liquid A was obtained, with a pressing rate of 66.8%.

[0045] (4) Add 1.1% by weight of preservatives (phenoxyethanol 0.9%, benzoic acid 0.12%, dehydroacetic acid 0.08%).

[0046] (5) Add appropriate amount of citric acid to adjust the pH to 5.5.

[0047] (6) Centrifuge (3000 rpm, 30 min), add 0.3% by weight of suspending agent (hydroxypropyl methylcellulose), stir well to obtain turmeric stock solution, and measure the solid content of the turmeric stock solution to be 2.78%.

[0048] Example 4 (1) Wash, dry and disinfect the turmeric.

[0049] (2) Weigh 600 g of detoxified turmeric, cut it into cubes, put it into a pressing bag, and place it into the press barrel.

[0050] (3) Set the pressing pressure range to 10-15 MPa and continue pressing for 20 min. Take the residue for the next pressing. Repeat the pressing for 3 times and collect the liquid. A total of 288 g of turmeric liquid A was obtained, with a pressing rate of 48.0%.

[0051] (4) Add 1.1% preservatives by weight (phenoxyethanol 0.9%, benzoic acid 0.12%, dehydroacetic acid 0.08%).

[0052] (5) Add appropriate amount of citric acid to adjust the pH to 5.5.

[0053] (6) Centrifuge (3000 rpm, 30 min), add 0.3% by weight of suspending agent (0.1% xanthan gum + 0.2% hydroxypropyl methylcellulose), stir well to obtain turmeric stock solution, and measure the solid content of the turmeric stock solution to be 3.03%.

[0054] Test Example 1 (Determination of polyphenols and flavonoids in turmeric stock solution) 1. Test steps (1) Determination of flavonoid content in turmeric stock solution Take 10 mg of rutin standard, dilute to 100 mL with 70% ethanol solution, and mix well. Prepare rutin standard solutions with concentrations of 20 ug / mL, 40 ug / mL, 60 ug / mL, 80 ug / mL, and 100 ug / mL, add 0.4 mL of 5% NaNO2 solution, shake well and let stand for 6 minutes, add 0.4 mL of 10% Al(NO3)3 solution, shake well and let stand for 6 minutes. Then absorb 4 mL of 4% NaOH and add it, let stand at room temperature for 15 minutes, and measure the absorbance at 510 nm. Draw a rutin standard curve based on the absorbance.

[0055] Accurately pipette 1.5 mL of turmeric stock solution into a 10 mL volumetric flask, add 5% NaNO2, 10% Al(NO3)3 and 4% NaOH as above, add distilled water to the mark, shake well, let stand at room temperature for 15 min, and measure the absorbance at a wavelength of 510 nm. Calculate the flavonoid content in the turmeric stock solution based on the measured absorbance and the above rutin standard curve.

[0056] (2) Determination of polyphenol content in turmeric stock solution Prepare gallic acid standard solutions with mass concentrations of 2, 3, 4, 5, 6, and 7 μg / mL, draw 0.05 mL of each, add 1.0 mL of Folin phenol reagent, add 4 mL of 7.5% Na2CO3 solution after 6 minutes, make up to 10 mL with deionized water, let stand for 60 minutes, and measure the absorbance at a wavelength of 765 nm. Draw a standard curve with mass concentration as the horizontal axis and absorbance as the vertical axis, and the regression equation of the standard curve is Y=0.133 X+0.0574, R 2 =0.998. Take the turmeric stock solution and measure its absorbance in the same way. Calculate the polyphenol content in the turmeric stock solution based on the measured absorbance and the standard curve regression equation.

[0057] 2. Turmeric stock solution test results Example 1: Total polyphenol content: 2.96±0.22 mg / ml; total flavonoid content: 2.08±0.07 mg / ml.

[0058] Example 2: Total polyphenol content: 2.65±0.11 mg / ml; total flavonoid content: 1.70±0.09 mg / ml.

[0059] Example 3: Total polyphenol content: 2.45±0.08 mg / ml; total flavonoid content: 1.60±0.13 mg / ml.

[0060] Example 4: Total polyphenol content: 2.55±0.04 mg / ml; total flavonoid content: 1.68±0.17 mg / ml.

[0061] Test Example 2 (Pesticide Residue Detection) The turmeric stock solution of Example 1 was subjected to GC-MS gas chromatography-mass spectrometry analysis. The analysis results are shown in Table 1.

[0062] Table 1 GC-MS gas chromatography-mass spectrometry analysis results of turmeric stock solution (pesticide residues)

[0063] The results in Table 1 show that the turmeric stock solution in Example 1 has no pesticide residue. The turmeric stock solutions in Examples 2 to 4 were subjected to the same analysis, and the results were the same as in Example 1. Therefore, the turmeric stock solutions of the present invention have no pesticide residue, which meets one of the conditions for plant extracts to be used as cosmetic raw materials.

[0064] The turmeric stock solutions used in the following test examples 3 to 5 are all the turmeric stock solutions prepared in Example 3.

[0065] Test Example 3 (mildness test) A 10% solution was prepared from turmeric stock solution and applied to CAM membrane for mildness test with 5 minutes as the end point. A 1% SLS aqueous solution was used as the positive control. The test method was derived from the chicken embryo chorioallantoic membrane test for eye irritation / corrosion of cosmetics (SN / T2329-2009).

[0066] The eye irritation scores of CAM membranes are shown in Table 2. The pictures of CAM membranes at the end point are shown in Table 2. Figure 1 shown.

[0067] Table 2 CAM membrane eye irritation score (scoring method: IS)

[0068] The formal test results are calculated from Table 2, IS = 0, which shows that the 10% turmeric stock solution is classified as non-irritating to the eyes.

[0069] Test Example 4 (Cytotoxicity Screening) The cultured cells were inoculated in a 96-well plate and placed in an incubator for 18-24 h until the cells were 80% confluent. The original culture medium was removed, and 100 μL of turmeric stock solution dilutions of different mass concentrations were added to each well and exposed in the incubator for 24±0.5 h. 20 μL of MTT solution was added to each well and incubated at 37°C for 3±0.5 h. Then the MTT solution was removed, 100 μL of DMSO was added to each well, and the absorbance value was measured at a wavelength of 570 nm after oscillation in the dark for 10-15 min.

[0070] Cytotoxicity test results are shown in Figure 3 The horizontal axis is the concentration of the turmeric stock solution dilution (where control is the blank control group), and the vertical axis is the cell activity. Figure 3 It can be seen that when the concentration is lower than 0.2006%, the cell survival rate is above 90%.

[0071] 0.100%, 0.050% and 0.025% were used as test concentrations for subsequent anti-inflammatory and soothing experiments.

[0072] Test Example 5 (Anti-inflammatory and soothing test) 1. Test steps: (1) HaCaT cells were routinely cultured by seeding the cells into 12-well plates and returning them to the incubator for 18-24 hours.

[0073] (2) Remove the culture plate and discard the original culture medium in the wells. Add DMEM culture medium to the negative control (NC), add DMEM culture medium containing LPS to the model control group (M), add DMEM culture medium containing LPS and different mass concentrations of turmeric stock solution to each well of the sample group (TA), and add DMEM culture medium containing LPS and dexamethasone to each well of the positive control group (PC). Each group has 3 replicate wells and cultured for 24±1 h.

[0074] (3) Collect the supernatant and store it at -80°C. Then measure the cytokines using ELISA kit.

[0075] 2. Test results: Experimental groups Figure 4 , the comparison results of IL-1α in different groups (Mean±SD) are shown in Figure 5 .

[0076] Depend on Figure 5 The results showed that the relative content of IL-1α in the turmeric stock solution at the three test concentrations was reduced to different levels compared with the model control group; the relative content of IL-1α in the turmeric stock solution sample at the 0.025% test concentration was significantly reduced compared with the model control group (p<0.05). In summary, the turmeric stock solution of the present invention has anti-inflammatory and soothing effects at the three test concentrations, among which the anti-inflammatory and soothing effect at the 0.025% mass concentration is the best.

[0077] The above disclosure is only the preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. A method for preparing an anti-inflammatory and soothing turmeric stock solution, characterized in that: The steps include: S1. Pre-treating turmeric to obtain turmeric to be squeezed; S2, squeezing the turmeric to be squeezed with a squeezer to obtain turmeric liquid A; S3, adding a preservative to the turmeric solution A and adjusting the pH, collecting and obtaining turmeric solution B; S4, centrifuging the turmeric solution B to obtain a supernatant; S5. Add a suspending agent to the upper layer liquid to obtain an anti-inflammatory and soothing turmeric stock solution.

2. The method for preparing the anti-inflammatory and soothing turmeric stock solution according to claim 1, characterized in that: The suspending agent includes one or both of xanthan gum and hydroxypropyl methylcellulose.

3. The method for preparing the anti-inflammatory and soothing turmeric stock solution as claimed in claim 1, characterized in that: The mass ratio of the suspending agent to the turmeric solution A is 0.1% to 1.0%; the solid content of the anti-inflammatory and soothing turmeric stock solution is 2.7% to 4.0%.

4. The method for preparing the anti-inflammatory and soothing turmeric stock solution as claimed in claim 1, characterized in that: The anti-inflammatory and soothing turmeric stock solution has a total polyphenol content of 2.4 to 3.0 mg / ml and a total flavonoid content of 1.6 to 2.1 mg / ml.

5. The method for preparing the anti-inflammatory and soothing turmeric stock solution as claimed in claim 1, characterized in that: The press described in step S2 uses a pressing plate as a pressing component; the pressing pressure described in step S2 includes 10 to 15 MPa; the single pressing time described in step S2 includes 10 to 30 minutes; the number of pressings described in step S2 includes 3 to 7 times; the centrifugal speed described in step S4 includes 1000 to 10000 rmp / min, and the time includes 30 minutes.

6. The method for preparing the anti-inflammatory and soothing turmeric stock solution as claimed in claim 1, characterized in that: The pretreatment in step S1 includes washing, drying, disinfecting and cutting in sequence; the preservative in step S3 includes a mixture of phenoxyethanol and sodium benzoate, or a mixture of phenoxyethanol, benzoic acid and dehydroacetic acid; the pH is adjusted to 5.5 in step S3; and step S5 includes: adding the suspending agent to the upper layer liquid, mixing evenly, and obtaining the anti-inflammatory and soothing turmeric stock solution.

7. The method for preparing the anti-inflammatory and soothing turmeric stock solution as claimed in claim 6, characterized in that: The slicing in step S1 includes dicing; the mass ratio of the preservative in step S3 to the turmeric liquid A in step S2 includes 1% to 1.1%; compared with the mass of the turmeric liquid A, the preservative includes a mixture of 0.8% by mass of phenoxyethanol and 0.2% by mass of sodium benzoate, or a mixture of 0.9% by mass of phenoxyethanol, 0.12% by mass of benzoic acid and 0.08% by mass of dehydroacetic acid; citric acid is used to adjust the pH in step S3; and the mixing method in step S5 includes stirring.

8. An anti-inflammatory and soothing turmeric stock solution obtained by the preparation method of the anti-inflammatory and soothing turmeric stock solution as claimed in claim 1.

9. An application of the anti-inflammatory and soothing turmeric stock solution as claimed in claim 8, characterized in that: Used to prepare preparations with anti-inflammatory and soothing effects.

10. The use according to claim 9, characterized in that The anti-inflammatory effect includes inhibiting the expression of inflammatory factor IL-1α by keratinocytes under external stimulation.

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