A black goji berry composition with uric acid-lowering effects

Abstract

This invention belongs to the field of novel food and health food formulation technology, specifically relating to a black goji berry composition with uric acid-lowering effects, comprising black goji berry freeze-dried powder and celery seed extract; the mass ratio of the black goji berry freeze-dried powder to the celery seed extract is 2.5–10:1. Through experiments on the effect of xanthine oxidase activity, it was determined that a ratio of black goji berry freeze-dried powder to celery seed extract of 5:1 showed a more effective xanthine oxidase inhibitory effect than using black goji berry freeze-dried powder or celery seed extract alone.

Description

Technical Field

[0001] This invention belongs to the field of food and health food technology, and specifically relates to a black goji berry composition with uric acid-lowering effects. Background Technology

[0002] Hyperuricemia is classified into primary and secondary hyperuricemia. Xanthine oxidase (XOD) is a key enzyme in purine metabolism, continuously catalyzing the oxidation of xanthine or hypoxanthine into uric acid. Xanthine oxidase is the crucial enzyme in the final step of uric acid synthesis; therefore, inhibiting XOD activity is a treatment for hyperuricemia. It is also considered a promising target for treating hyperuricemia and gout.

[0003] Black goji berries are the dried fruit of the wild plant *Lycium ruthenicum* Murr., belonging to the Solanaceae family. The berries are spherical, turning purplish-black when ripe. They are non-toxic, sweet, and neutral in nature, possessing benefits such as improving eyesight, nourishing the liver, and tonifying the kidneys and replenishing essence. Studies have shown that long-term intake of black goji berry anthocyanin extract can effectively inhibit the rise of serum uric acid levels and exhibit preventative and therapeutic effects on acute gouty arthritis. Celery seeds are the seeds of celery plants, containing various physiologically active substances, mainly including volatile oil compounds, unsaturated fatty acids, flavonoids, butylphthalides, amino acids, dietary fiber, coumarins and their derivatives, etc., and have physiological functions such as lowering blood pressure and blood lipids. Angelica dahurica has analgesic and dampness-removing effects. It is a traditional Chinese medicine that is commonly used for rheumatic pain and can be used as both food and medicine. In vitro experiments have shown that the water extract of Angelica dahurica has a strong inhibitory effect on xanthine oxidase [Wang Xiaomei, Zhao Yan, Liu Dan, et al. Inhibitory effect of three Chinese medicine extracts on xanthine oxidase [J]. Amino Acids and Biological Resources, 2013, 35(1):57-60].

[0004] The main active ingredient in black goji berries is anthocyanin, a water-soluble natural plant pigment that is readily soluble in polar solvents such as methanol, dilute alkalis, and dilute acids. It is relatively stable under acidic conditions but degrades and loses its activity under alkaline conditions. To address the stability issue of anthocyanins in black goji berries, we prepared a black goji berry composition. In vitro studies showed that a combination of black goji berry and celery seed extract in a certain proportion exhibited a more effective inhibitory effect on xanthine oxidase. Summary of the Invention

[0005] In view of the drawback of the easy degradation of anthocyanins in black goji berries in existing technologies, the purpose of this invention is to provide a black goji berry composition that can effectively improve the stability of anthocyanins, significantly lower blood uric acid, and has no toxic side effects. Through experiments on the effect of xanthine oxidase activity, it was determined that a ratio of 5:1 between freeze-dried black goji berry powder and celery seed extract showed a more effective xanthine oxidase inhibitory effect than using freeze-dried black goji berry powder or celery seed extract alone.

[0006] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0007] A black goji berry composition with uric acid-lowering effects includes black goji berry freeze-dried powder and celery seed extract; the mass ratio of the black goji berry freeze-dried powder to the celery seed extract is 2.5 to 10:1.

[0008] Studies have shown that celery seeds contain 50 times more apigenin than celery, which has a good effect on lowering blood uric acid. In in vitro experiments, this invention found that the combination of freeze-dried black goji berry powder and celery seed extract exhibited strong inhibitory activity against XOD.

[0009] Preferably, the mass ratio of the freeze-dried black goji berry powder to the celery seed extract is 5:1.

[0010] When the mass ratio of the freeze-dried black goji berry powder to the celery seed extract is 5:1, it exhibits significantly better xanthine oxidase (XOD) inhibition than using the freeze-dried black goji berry powder alone.

[0011] Preferably, it also includes ascorbic acid, sodium ascorbate, maltodextrin, xanthan gum, and acidulants.

[0012] Preferably, the raw materials include the following parts by weight: 400.0 parts of the freeze-dried black goji berry powder, 80.0 parts of the celery seed extract, 1.0 part of the mixture of ascorbic acid and sodium ascorbate in a mass ratio of 3:1, 100.0 parts of the maltodextrin, 10.0 parts of the xanthan gum, and 0.3 to 3.5 parts of the acid agent.

[0013] Preferably, the acidic agent includes one or more of citric acid and malic acid.

[0014] A method for preparing the above-mentioned black goji berry composition with uric acid-lowering effect includes the following steps:

[0015] A. Mix the maltodextrin with the xanthan gum, add purified water, heat, stir until completely dissolved, cool, and obtain a first mixture;

[0016] B. Add the black goji berry freeze-dried powder and the celery seed extract to the first mixture, and continue stirring until completely dissolved to obtain the second mixture;

[0017] C. Add the mixture of ascorbic acid and sodium ascorbate to the second mixture while stirring to obtain a mixed solution;

[0018] D. Sterilize the mixed solution to obtain a sterilized mixed solution;

[0019] E. Freeze-dry the sterilized mixed solution to obtain the black goji berry composition.

[0020] Preferably, step A includes: mixing the maltodextrin and xanthan gum, adding purified water, heating to 70-80°C, stirring until completely dissolved, and cooling to 20-30°C to obtain the first mixture; step C includes: adding a mixture of ascorbic acid and sodium ascorbate in a mass ratio of 3:1 to the second mixture, adding an acid while stirring, adjusting the pH to 4-6, and continuing to stir for 30 minutes to obtain the mixed solution; in step D, the sterilization includes filter membrane sterilization.

[0021] Preferably, the pore size of the filter membrane for sterilization includes 0.2 to 0.45 μm.

[0022] The application of the above-mentioned black goji berry composition with uric acid-lowering effect in the preparation of preparations for treating or preventing hyperuricemia.

[0023] Preferably, its use in the preparation of formulations that inhibit xanthine oxidase.

[0024] Compared with the prior art, implementing the present invention has the following beneficial effects:

[0025] 1. Black goji berry freeze-dried powder and celery seed extract, when in a certain ratio, have a synergistic effect in inhibiting xanthine oxidase.

[0026] 2. This invention has discovered a composition of freeze-dried black goji berry powder and celery seed extract: which can be used to prepare drugs that lower uric acid. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments, so that those skilled in the art can better understand and implement this invention. However, the embodiments are not intended to limit the invention. Modifications or substitutions made to the methods, steps, or conditions of this invention without departing from the spirit and substance of this invention are all within the scope of this invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0028] Example 1

[0029] A method for preparing a black goji berry composition with uric acid-lowering effects includes the following steps:

[0030] Weigh out 100.0g of maltodextrin and 10.0g of xanthan gum, mix them, add purified water, heat to 75℃, stir until completely dissolved, and cool to 25℃. Then, weigh out 400.0g of freeze-dried black goji berry powder and 80.0g of celery seed extract, continue stirring until completely dissolved, then add 1.0g of a mixture of ascorbic acid and sodium ascorbate in a 3:1 ratio. While stirring, add malic acid or citric acid to adjust the pH to 4-6, and continue stirring for 30 minutes to obtain a mixed solution. Sterilize the dissolved mixed solution by passing it through a filter membrane with a pore size of 0.22μm to obtain a sterilized mixed solution. Prepare the freeze-dried black goji berry composition powder by freeze-drying.

[0031] Example 2

[0032] The effects of xanthine oxidase activity inhibition were compared using black goji berry freeze-dried powder and celery seed extract alone, as well as in combination with different proportions of these mixtures, through a xanthine oxidase activity inhibition assay. The specific steps included:

[0033] (1) Weigh 0.2g of NaOH and dissolve it in 50mL of distilled water to prepare a NaOH solution with a concentration of 0.1mol / L.

[0034] (2) Preparation of phosphate buffer (0.2 mol / L, pH = 7.4, PB): Weigh 9.742 g of K₂HPO₄·3H₂O, 0.995 g of KH₂PO₄, and 0.09305 g of EDTA-2Na, dissolve them in distilled water, adjust the pH to 7.4 with 0.1 mol / L NaOH solution, and dilute to a 250 mL volumetric flask. Then pour the solution into a reagent bottle to prepare a 0.2 mol / L phosphate buffer solution. Store at 4℃ for an extended period.

[0035] (3) Xanthine solution: Accurately weigh xanthine, dissolve it completely in 0.1 mol / L NaOH, and then dilute it with phosphate buffer.

[0036] (4) Xanthine oxidase solution: Dilute XOD with a certain amount of phosphate buffer.

[0037] (5) Sample solution: Weigh a certain amount of black goji berry freeze-dried powder and celery seed extract respectively, and prepare mixtures with different mass ratios (black goji berry freeze-dried powder: celery seed extract is 2.5:1, 5:1, 10:1). After dissolving the mixture in water, take the supernatant and pass it through a 0.45μm membrane.

[0038] (6) Add 98 μL of phosphate buffer to a 96-well plate, then add different concentration ratios of the composition and 0.05 mmol / L xanthine solution, incubate at 37°C for 5 min, and then add 1 μL of 3 U / L enzyme solution to the microplate. Place the plate in a microplate reader for incubation at 37°C and detection. The absorbance detected by the microplate reader at 0 min is OD4, and the absorbance detected by the microplate reader after 30 min is OD3. The reaction system without inhibitor and with other solvents is the control group. The absorbance detected at 0 min is OD1. The reaction is carried out at 37°C. The absorbance OD4 of the drug-treated group is measured at 294 nm after 30 min. Each group of experiments is repeated in triplicate. The formula for calculating the inhibition rate is as follows:

[0039] Inhibition rate (%) = (1 - OD4 - OD3 / OD2 - OD1) × 100

[0040] The inhibition rate results are shown in Table 1:

[0041] Table 1. Test results for each group

[0042]

[0043] The test results were statistically analyzed using SPSS software, and one-way ANOVA was used. P < 0.05 was considered statistically significant. In the multiple comparisons (LSD), compared with the use of black goji berry freeze-dried powder or celery seed extract alone, the ratio of black goji berry freeze-dried powder to celery seed extract of 2.5:1 to 10:1 significantly improved the inhibitory effect on xanthine oxidase (P < 0.05), and the inhibitory effect on purine oxidase was best when the mass ratio of black goji berry freeze-dried powder to celery seed extract was 5:1 (P < 0.01).

[0044] Comparative Example 1

[0045] The study compared the effects of using Angelica dahurica extract alone versus a mixture of black goji berry and Angelica dahurica extract on enzyme activity inhibition using a xanthine oxidase activity inhibition assay. The specific steps included:

[0046] (1) Weigh 0.2g of NaOH and dissolve it in 50mL of distilled water to prepare a NaOH solution with a concentration of 0.1mol / L.

[0047] (2) Preparation of phosphate buffer (0.2 mol / L, pH = 7.4, PB): Weigh 9.742 g of K₂HPO₄·3H₂O, 0.995 g of KH₂PO₄, and 0.09305 g of EDTA-2Na, dissolve them in distilled water, adjust the pH to 7.4 with 0.1 mol / L NaOH solution, and dilute to a 250 mL volumetric flask. Then pour the solution into a reagent bottle to prepare a 0.2 mol / L phosphate buffer solution. Store at 4℃ for an extended period.

[0048] (3) Xanthine solution: Accurately weigh xanthine, dissolve it completely in 0.1 mol / L NaOH, and then dilute it with phosphate buffer.

[0049] (4) Xanthine oxidase solution: Dilute XOD with a certain amount of phosphate buffer.

[0050] (5) Sample solution: Weigh the black goji berry composition, angelica extract, and black goji berry extract and angelica extract composition (mass ratio 5:1) prepared in Example 1, dissolve the mixture in water, take the supernatant, and pass it through a 0.45 μm membrane.

[0051] (6) Add 98 μL of phosphate buffer to a 96-well plate, then add different concentration ratios of the composition and 0.05 mmol / L xanthine solution, incubate at 37°C for 5 min, and then add 1 μL of 3 U / L enzyme solution to the microplate. Place the plate in a microplate reader for incubation at 37°C and detection. The absorbance detected by the microplate reader at 0 min is OD4, and the absorbance detected by the microplate reader after 30 min is OD3. The reaction system without inhibitor and with other solvents is the control group. The absorbance detected at 0 min is OD1. The reaction is carried out at 37°C. The absorbance OD4 of the drug-treated group is measured at 294 nm after 30 min. Each group of experiments is repeated in triplicate. The formula for calculating the inhibition rate is as follows:

[0052] Inhibition rate (%) = (1 - OD4 - OD3 / OD2 - OD1) × 100

[0053] The inhibition rate results are shown in Table 2:

[0054] Table 2. Test results for each group

[0055]

[0056] Note: The black goji berry composition is the one prepared in Example 1, namely, black goji berry freeze-dried powder / celery seed extract (5:1).

[0057] The test results were statistically processed using SPSS software, and one-way ANOVA was used. P < 0.05 was considered statistically significant. In the multiple comparisons (LSD) combined with Table 1, it was surprisingly found that although Angelica dahurica extract showed stronger inhibitory effects on xanthine oxidase compared to celery seed extract and Angelica dahurica extract alone, when combined with freeze-dried black goji berry powder, the combination of freeze-dried black goji berry powder and celery seed extract, at the same ratio, unexpectedly exhibited superior inhibitory effects on xanthine oxidase compared to different groups of freeze-dried black goji berry powder / Angelica dahurica extract.

[0058] The above-disclosed embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. Therefore, equivalent variations made in accordance with the claims of the present invention are still within the scope of the present invention.

Claims

1. A black goji berry composition with uric acid-lowering effects, characterized in that, The active ingredients are freeze-dried black goji berry powder and celery seed extract; the mass ratio of the freeze-dried black goji berry powder to the celery seed extract is 5:

1.

2. The black goji berry composition with uric acid-lowering effect as described in claim 1, characterized in that, It also includes ascorbic acid, sodium ascorbate, maltodextrin, xanthan gum, and acidifiers.

3. The black goji berry composition with uric acid-lowering effect as described in claim 2, characterized in that, The raw materials include the following parts by weight: 400.0 parts of the freeze-dried black goji berry powder, 80.0 parts of the celery seed extract, 1.0 part of the mixture of ascorbic acid and sodium ascorbate in a mass ratio of 3:1, 100.0 parts of the maltodextrin, 10.0 parts of the xanthan gum, and 0.3 to 3.5 parts of the acid agent.

4. The black goji berry composition with uric acid-lowering effect as described in claim 2, characterized in that, The acidic agent includes one or more of citric acid and malic acid.

5. A method for preparing a black goji berry composition with uric acid-lowering effect as described in claim 2, characterized in that, Includes the following steps: A. Mix the maltodextrin with the xanthan gum, add purified water, heat, stir until completely dissolved, cool, and obtain a first mixture; B. Add the black goji berry freeze-dried powder and the celery seed extract to the first mixture, and continue stirring until completely dissolved to obtain the second mixture; C. Add the mixture of ascorbic acid and sodium ascorbate to the second mixture while stirring to obtain a mixed solution; D. Sterilize the mixed solution to obtain a sterilized mixed solution; E. Freeze-dry the sterilized mixed solution to obtain the black goji berry composition.

6. The preparation method according to claim 5, characterized in that, Step A includes: mixing the maltodextrin and xanthan gum, adding purified water, heating to 70-80°C, stirring until completely dissolved, and cooling to 20-30°C to obtain the first mixture; Step C includes: adding a mixture of ascorbic acid and sodium ascorbate in a mass ratio of 3:1 to the second mixture, adding an acid while stirring, adjusting the pH to 4-6, and continuing to stir for 30 minutes to obtain the mixed solution; In Step D, the sterilization includes filter membrane sterilization.

7. The preparation method according to claim 6, characterized in that, The pore size of the filter membrane used for sterilization includes 0.2~0.45μm.

8. The application of a black goji berry composition with uric acid-lowering effect as described in claim 1, characterized in that, Application in the preparation of formulations for the treatment or prevention of hyperuricemia.

9. The application as described in claim 8, characterized in that, Application in the preparation of formulations that inhibit xanthine oxidase.

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