Functional hydrogen water formula capable of promoting uric acid excretion of gout and preparation technology of functional hydrogen water formula

Abstract

The invention discloses a functional hydrogen water formula capable of promoting uric acid excretion of gout, the functional hydrogen water formula comprises the following components: hydrogen, purified water, sodium bicarbonate, a magnesium ion compound and a vitamin C derivative, the dissolution concentration of the hydrogen in water is 0.8-2.0 mg / L, the addition amount of the sodium bicarbonate is 0.5-3.0 g / L, the concentration of the magnesium ion compound is 10-50 mg / L in terms of magnesium element, the concentration of the vitamin C derivative is 10-50 mg / L in terms of magnesium element, and the concentration of the vitamin C derivative is 0.5-1.0 mg / L. The addition amount of the vitamin C derivative is 0.1-1.0 g / L. The functional hydrogen water can accurately promote uric acid excretion, the serum uric acid (SUA) is reduced by 43.9% (216.3 + / -20.7 [mu] mol / L) compared with a model group and the urinary uric acid (UUA) is increased by 90.7% (238.9 + / -19.8 [mu] mol / 24h) in 24 hours after the functional hydrogen water intervenes in a hyperuricemia rat for 21 days, and the functional hydrogen water is superior to common hydrogen water and can be comparable to benzbromarone. The mechanism is as follows: sodium bicarbonate adjusts the pH value to 7.5-8.5 to help to dissolve urate, magnesium + hydrogen down-regulates URAT1 (reduced by 46.5%), up-regulates GLUT9 (increased by 93.1%) to reduce uric acid reabsorption, and also improves the activity of liver uricase (increased by 61.1%) to assist decomposition, so that a multi-channel excretion promoting system is formed.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of hydrogen water preparation, in particular to a functional hydrogen water formula capable of promoting uric acid excretion in gout and a preparation technology thereof. BACKGROUND

[0002] Gout, as a common metabolic disease, is closely related to the disorder of uric acid metabolism and the excretion disorder of uric acid in the body. In recent years, due to the influence of factors such as changes in dietary structure and changes in lifestyle, the incidence rate has been continuously increasing, and the patient population is gradually becoming younger. In clinical practice, promoting uric acid excretion is the core intervention direction for controlling gout attacks and improving patient prognosis. The existing intervention methods mainly include drug treatment and dietary conditioning: although drugs (such as benzbromarone) can quickly promote uric acid excretion, long-term use may cause side effects such as gastrointestinal discomfort, liver and kidney function damage, and drug dependence; although dietary conditioning (such as low-purine diet and increasing water intake) is highly safe, it is slow-acting and difficult to meet the rapid intervention needs of patients with acute attack or hyperuricemia, and there is an urgent need for functional drinks with high safety and strong targeting to assist in improving uric acid excretion.

[0003] Hydrogen water has gradually attracted attention in the field of metabolic disease auxiliary conditioning due to its good antioxidant and anti-inflammatory properties, but its existing technology has significant defects and is difficult to adapt to the uric acid excretion needs of gout patients: on the one hand, the formula design has limitations, ordinary functional hydrogen water mostly only takes "hydrogen gas + purified water" as the core ingredient, with single function, lacking of synergistic ingredients for uric acid excretion - without adding sodium bicarbonate that can adjust the pH value of body fluid and promote the dissolution of uric acid salt, without adding magnesium ion compounds that can activate uric acid transporters and assist the kidney in metabolizing uric acid, and without introducing vitamin C derivatives that can protect the stability of hydrogen gas and enhance antioxidant effect, resulting in that it can only play a basic antioxidant role and cannot effectively target promote uric acid excretion; at the same time, the hydrogen gas solubility concentration of some hydrogen water fluctuates greatly, and the concentration is too low, which is insufficient, and too high, which is easy to cause poor drinking experience due to hydrogen gas escaping, and lacks scientific matching of trace elements such as zinc ions, further limiting the functional extension.

[0004] On the other hand, the preparation technology has many shortcomings: first, the pretreatment of purified water is not complete, most of the processes do not use vacuum degassing or nitrogen replacement method to remove dissolved oxygen (the concentration of dissolved oxygen is often higher than 0.5 mg / L), the residual oxygen is easy to react with hydrogen, which leads to rapid attenuation of hydrogen concentration, and the conductivity (often higher than 10 μS / cm) is not controlled by multiple processes such as quartz sand filtration, activated carbon adsorption and reverse osmosis, and the impurities in the water may affect the subsequent component solubility; second, the hydrogen dissolving efficiency is low and unstable, the micro-nano bubble generator (bubble diameter is more than 500 nm) is not used, the aeration rate and temperature and pressure control are not proper, which makes it difficult to maintain the hydrogen solubility concentration at 0.8-2.0 mg / L, and some processes do not introduce ultrasonic treatment or use continuous ultrasonic method, which further reduces the dispersion and stability of hydrogen; third, the component adding sequence and process parameters are unreasonable, the adding sequence is not planned according to the solubility and reaction characteristics of the components (such as adding sodium bicarbonate to adjust pH first, then adding magnesium ion and vitamin C derivative), which is easy to cause the oxidation of vitamin C derivative and makes the components not mixed evenly due to improper stirring speed and time control; fourth, the filling and storage process is not perfect, the filling is not carried out in a hundred-level clean workshop, the 0.22 μm ceramic membrane filtration is not used for sterilization, the sealing container is mostly ordinary plastic bottle (not aluminum composite membrane or 304 stainless steel tank) and the inner wall is not passivated, which causes the residual oxygen content in the container to be higher than 0.1%, the hydrogen is easy to escape, and the light shielding and low temperature (2-10℃) storage measures are missing, which further shortens the product shelf life and affects the function stability.

[0005] In summary, the existing intervention means for gout uric acid excretion have the problem of difficult to balance safety and effectiveness, and functional hydrogen water has potential, but there are obvious shortcomings in formula synergy and preparation technology stability, which cannot meet the needs of gout patients for safe and efficient functional drinks for promoting uric acid excretion, therefore, it is a technical problem to be solved in the current field to develop a functional hydrogen water with scientific formula, stable preparation and targeted promotion of gout uric acid excretion. SUMMARY

[0006] (I) Technical problems solved

[0007] In view of the shortcomings of the prior art, the present application provides a functional hydrogen water formula and its preparation technology for promoting gout uric acid excretion, which solves the problems in the above background art.

[0008] (II) Technical solutions

[0009] To achieve the above object, the present application provides the following technical scheme: a functional hydrogen water formula capable of promoting uric acid excretion of gout, which is composed of hydrogen, purified water, sodium bicarbonate, magnesium ion compound and vitamin C derivative, wherein the solubility concentration of hydrogen in water is 0.8-2.0 mg / L, the addition amount of sodium bicarbonate is 0.5-3.0 g / L, the concentration of magnesium ion compound in terms of magnesium element is 10-50 mg / L, and the addition amount of vitamin C derivative is 0.1-1.0 g / L.

[0010] Preferably, the magnesium ion compound is at least one selected from magnesium chloride, magnesium sulfate and magnesium gluconate.

[0011] Preferably, the vitamin C derivative is sodium ascorbate phosphate or ascorbyl palmitate.

[0012] Preferably, 0.01-0.1 g / L of zinc ion compound is further included, and the zinc ion compound is selected from zinc sulfate or zinc gluconate.

[0013] A preparation technology of functional hydrogen water capable of promoting uric acid excretion of gout, for preparing the formula of any one of claims 1-4, characterized by comprising the following steps:

[0014] (1) pretreating the purified water: treating the purified water by vacuum degassing method at a pressure of-0.08 to-0.1 MPa for 20-40 minutes, or treating the purified water by nitrogen replacement method by passing nitrogen gas at a flow rate of 0.5-2 L / min for 30-60 minutes to remove dissolved oxygen in the water to a concentration of≤0.5 mg / L;

[0015] (2) contacting the pretreated purified water with hydrogen gas through a micro-nano bubble generator in a sealed reaction kettle, controlling the bubble diameter to be 50-500 nm, the aeration rate to be 1-5 L / h, the reaction pressure to be maintained at 0.1-0.5 MPa, and the temperature to be controlled at 2-15℃, and continuously contacting for 60-120 minutes to dissolve the hydrogen gas to a concentration of 0.8-2.0 mg / L to obtain hydrogen-rich water;

[0016] (3) transferring the hydrogen-rich water into a stirring tank, stirring at a speed of 300-500 rpm and adding sodium bicarbonate at a temperature of 20-30℃, dissolving and adjusting the pH to 7.0-8.0; then adding the magnesium ion compound and continuing to stir for 5-10 minutes; then adding the vitamin C derivative and maintaining the stirring for 10-20 minutes, and finally controlling the pH value of the solution to be 7.5-8.5;

[0017] (4) The functional hydrogen water is filtered and sterilized by a 0.22 μm ceramic membrane, and then filled into a sealed container in a hundred-level clean workshop at a speed of 50-100 ml / s. After filling, the container is sealed by heat sealing or a screw cap, and the residual oxygen content in the container is ensured to be ≤0.1%, and then the container is stored in a dark environment at 2-10°C.

[0018] Preferably, the step (2) further comprises ultrasonic treatment of the dissolved hydrogen-rich water: ultrasonic treatment is performed at a frequency of 20-40 kHz and a power of 100-300 W for 5-15 minutes, and the temperature is kept ≤15°C during the ultrasonic treatment.

[0019] Preferably, in the step (3), if the zinc ion compound is added, the zinc ion compound is added after the vitamin C derivative is added, and the stirring is continued for 5-10 minutes until the zinc ion compound is completely dissolved.

[0020] Preferably, in the step (4), the sealed container is an aluminum composite film packaging bag or a food-grade 304 stainless steel tank, and the inner wall of the container is passivated.

[0021] Preferably, in the step (1), the purified water needs to be filtered through quartz sand, adsorbed by activated carbon, and treated by reverse osmosis so that the conductivity is ≤10 μS / cm.

[0022] Preferably, the ultrasonic treatment is intermittent operation, that is, working for 30 seconds and pausing for 10 seconds, and the cycle is repeated until the set treatment time is reached.

[0023] (Three) Beneficial effects

[0024] Compared with the prior art, the present application provides a functional hydrogen water formula capable of promoting uric acid excretion and a preparation technology thereof, and has the following beneficial effects:

[0025] 1. High-efficiency targeted uric acid excretion promotion effect: The functional hydrogen water can accurately promote uric acid excretion: after 21 days of intervention on high uric acid rats, the serum uric acid (SUA) is reduced by 43.9% (216.3±20.7 μmol / L) compared with the model group, and the 24-hour urine uric acid (UUA) is increased by 90.7% (238.9±19.8 μmol / 24h), which is better than ordinary hydrogen water and comparable to benzbromarone. The mechanism is: sodium bicarbonate adjusts pH 7.5-8.5 to help urate dissolution, magnesium + hydrogen down-regulates URAT1 (decreases by 46.5%) and up-regulates GLUT9 (increases by 93.1%) to reduce uric acid reabsorption, and also increases the activity of liver uricase (increases by 61.1%) to assist decomposition, forming a multi-pathway excretion system.

[0026] 2. Excellent ingredient and storage stability effect: With optimized preparation technology, the hydrogen water ingredient is stable and easy to store for a long period of time: dissolution stability: the purified water is treated in multiple steps (conductivity ≤ 10 μS / cm), oxygen is controlled to ≤ 0.5 mg / L by degassing / nitrogen replacement, and the hydrogen concentration is stabilized at 0.8-2.0 mg / L by micro-nano bubbles + intermittent ultrasound, for example, 1.2 mg / L of hydrogen at 5°C for 7 days in the dark (decrease of 14.3%); mixing stability: add in the order of "sodium bicarbonate → magnesium → C (add zinc after)" and stir at 300-500 rpm, and the retention rate of C is ≥ 95%, for example, 1.5 mg / L of hydrogen at 8°C for 10 days in the dark and clear.

[0027] 3. High safety and no side effects: biological safety: after 21 days of drinking for high uric acid rats, the serum Cr (55-70 μmol / L) and BUN (4.5-6.0 mmol / L) are normal, and there is no difference with the blank group; the indicators of the benzbromarone group are significantly increased, and there is a kidney burden; ingredient technology safety: the raw materials are food grade, the zinc is added at 0.01-0.1 g / L, the preparation is sterilized by 0.22 μm (≥ 99.9%), and the hundred-level filling is sealed to control the oxygen to ≤ 0.1%, which meets the food grade standard. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of the present application.

[0029] Example 1: Preparation of functional hydrogen water of basic formula

[0030] 1. Formula composition (based on 1 L of functional hydrogen water)

[0031] Purified water: 997.15 g (conductivity ≤ 10 μS / cm)

[0032] Hydrogen: dissolved concentration 1.4 mg / L

[0033] Sodium bicarbonate: 1.5 g

[0034] Magnesium ion compound: magnesium chloride (25 mg of magnesium element, corresponding to 0.13 g of magnesium chloride)

[0035] Vitamin C derivative: sodium ascorbate phosphate 0.5 g

[0036] No zinc ion compound is added

[0037] 2. Preparation steps

[0038] (1) Purified water pretreatment: The purified water was sequentially subjected to quartz sand filtration (to remove suspended solids), activated carbon adsorption (to remove organic matter and residual chlorine), and reverse osmosis treatment (to reduce the electrical conductivity to 8 μS / cm); then vacuum degassing was performed at a pressure of -0.09 MPa for 30 minutes, and the dissolved oxygen concentration in the water was detected to be 0.3 mg / L, meeting the requirement of ≤0.5 mg / L.

[0039] (2) Preparation of hydrogen-rich water: The pretreated purified water was transferred into a sealed reaction kettle, hydrogen gas was introduced through a micro-nano bubble generator, the bubble diameter was controlled to be 200 nm, the aeration rate was 3 L / h, the reaction pressure was maintained at 0.3 MPa, and the temperature was 8°C; after 90 minutes of continuous contact, the hydrogen gas solubility concentration was detected to be 1.4 mg / L; then ultrasonic treatment was performed at a frequency of 25 kHz and a power of 200 W in an intermittent mode (30 seconds of operation and 10 seconds of pause), and the total treatment time was 10 minutes; during the process, the temperature was maintained at 12°C to 15°C, and hydrogen-rich water was obtained.

[0040] (3) Mixing of functional ingredients: The hydrogen-rich water was transferred into a stirring tank, the temperature was controlled to be 25°C, stirring was performed at a speed of 400 rpm, and 1.5 g of sodium bicarbonate was added; after dissolution, the pH value was detected to be 7.5; then 0.13 g of magnesium chloride was added, and stirring was continued for 8 minutes until complete dissolution; then 0.5 g of sodium ascorbate phosphate was added, and stirring was maintained at a speed of 400 rpm for 15 minutes, and the pH value was finally adjusted to 8.0.

[0041] (4) Filling and storage: The mixed functional hydrogen water was filtered and sterilized through a 0.22 μm ceramic membrane (sterilization rate ≥99.9%), and was filled into aluminum composite film packaging bags (the inner wall was passivated) at a speed of 80 ml / s in a hundred-level clean workshop; after filling, heat sealing was performed, and the residual oxygen content in the bag was detected to be 0.08% to 0.1%; finally, the product was stored in the dark at 5°C, and after 7 days of standing, the hydrogen gas concentration was still maintained at 1.2 mg / L, and the solution was clear and transparent without precipitation.

[0042] Example Two: Preparation of a Functional Hydrogen Water Containing Zinc Ions

[0043] 1. Formula composition (based on 1 L of functional hydrogen water)

[0044] Purified water: 995.37 g (electrical conductivity ≤10 μS / cm)

[0045] Hydrogen gas: solubility concentration 1.8 mg / L

[0046] Sodium bicarbonate: 2.5 g

[0047] Magnesium ion compound: magnesium sulfate (40 mg of magnesium element, corresponding to 0.24 g of magnesium sulfate)

[0048] Vitamin C derivative: Ascorbic acid palmitate 0.8 g

[0049] Zinc ion compound: Zinc sulfate (0.05 g of zinc element, corresponding to 0.13 g of zinc sulfate)

[0050] 2. Preparation steps

[0051] (1) Purified water pretreatment: The purified water was filtered through quartz sand, adsorbed by activated carbon, and treated by reverse osmosis, with an electrical conductivity of 6 μS / cm; nitrogen was introduced at a flow rate of 1.2 L / min for 45 min using the nitrogen replacement method, and the dissolved oxygen concentration was detected to be 0.4 mg / L.

[0052] (2) Preparation of hydrogen-rich water: The purified water was transferred into a sealed reaction kettle, and hydrogen was introduced through a micro-nano bubble generator, with a bubble diameter of 300 nm and an aeration rate of 4 L / h, a reaction pressure of 0.4 MPa and a temperature of 12°C, and the hydrogen-rich water was obtained by continuous contact for 110 min without ultrasonic treatment, with a hydrogen dissolved concentration of 1.8 mg / L.

[0053] (3) Mixing of functional ingredients: The temperature of the stirring tank was controlled at 28°C, and the hydrogen-rich water was stirred at a speed of 500 rpm and 2.5 g of sodium bicarbonate was added, and after dissolution, the pH value was 7.8; 0.24 g of magnesium sulfate was added and stirred for 10 min until dissolved; then 0.8 g of ascorbic acid palmitate was added and stirred for 20 min; finally, 0.13 g of zinc sulfate was added and continued to stir for 7 min until completely dissolved, and the final pH value was adjusted to 8.3.

[0054] (4) Filling and storage: After sterilization by 0.22 μm ceramic membrane filtration, it was filled into food-grade 304 stainless steel tanks (inner wall passivation treatment) at a speed of 100 ml / s in a hundred-level clean workshop; sealed with a screw cap, and the residual oxygen content in the tank was detected to be 0.09%; stored in the dark at 8°C, and after 10 days, the hydrogen concentration was 1.5 mg / L, and the solution had no odor and no stratification.

[0055] Example Three: Preparation of functional hydrogen water with low concentration of ingredients

[0056] 1. Formula composition (based on 1 L of functional hydrogen water)

[0057] Purified water: 998.82 g (electrical conductivity ≤ 10 μS / cm)

[0058] Hydrogen: dissolved concentration 0.9 mg / L

[0059] Sodium bicarbonate: 0.8 g

[0060] Magnesium ion compound: Magnesium gluconate (15 mg of magnesium element, corresponding to 0.21 g of magnesium gluconate)

[0061] Vitamin C derivative: sodium ascorbate phosphate 0.2 g

[0062] Zinc ion compound: zinc gluconate (0.02 g of zinc element, corresponding to 0.08 g of zinc gluconate mass)

[0063] 2. Preparation step

[0064] (1) Purified water pretreatment: the purified water was filtered through quartz sand, adsorbed by activated carbon, and treated by reverse osmosis, and the conductivity was 5 μS / cm; vacuum degassing was used, and the dissolved oxygen concentration was reduced to 0.2 mg / L under the pressure of-0.08 MPa for 25 minutes.

[0065] (2) Preparation of hydrogen-rich water: in a sealed reaction kettle, the micro-nano bubble generator controlled the bubble diameter to be 80 nm and the aeration rate to be 1.5 L / h, the reaction pressure was 0.15 MPa and the temperature was 5℃, the contact time was 65 minutes, and the hydrogen gas dissolved concentration was 0.9 mg / L; ultrasonic treatment was performed, and the frequency was 40 kHz and the power was 100 W, intermittent operation (working for 30 seconds and pausing for 10 seconds), the total treatment time was 5 minutes, and the temperature was maintained at 4℃.

[0066] (3) Mixing of functional ingredients: the temperature of the stirring tank was 22℃, 0.8 g of sodium bicarbonate was added and stirred at the speed of 300 rpm, and after dissolution, the pH value was 7.2; 0.21 g of magnesium gluconate was added and stirred for 5 minutes; then 0.2 g of sodium ascorbate phosphate was added and stirred for 10 minutes; finally, 0.08 g of zinc gluconate was added and stirred for 5 minutes, and the final pH value was adjusted to 7.6.

[0067] (4) Filling and storage: after sterilization by 0.22 μm ceramic membrane filtration, it was filled into food-grade 304 stainless steel tanks (inner wall passivated) at the speed of 50 ml / s; the residual oxygen content was 0.07% after screw cap sealing; it was stored in the dark at 2℃, and after 15 days, the hydrogen concentration was 0.8 mg / L, which met the stability requirements of functional hydrogen water.

[0068] Experimental example: effect of functional hydrogen water on uric acid excretion of hyperuricemia rats

[0069] I. Purpose of the experiment

[0070] The serum uric acid reducing effect, uric acid excretion promoting effect and safety of the functional hydrogen water (prepared based on the formula of Example One) which can promote uric acid excretion of gout according to the application were verified, the intervention differences of ordinary hydrogen water and positive drugs were compared, and the effectiveness and advantages of the functional hydrogen water in promoting uric acid excretion were determined.

[0071] II. Experimental materials

[0072] Experimental animals: 40 SPF male SD rats, weighing 220-250 g, purchased from an experimental animal center, the feeding environment temperature is 22-25 DEG C, humidity is 50-60 %, 12h light-dark cycle, free food and water (ordinary feed + purified water), after 7 days of adaptation feeding, the experiment is started.

[0073] Reagents and instruments: potassium oxonate (uric acid production inhibitor, used for constructing a high uric acid model), uricase method serum / urine uric acid detection kit, creatinine (Cr) detection kit, urea nitrogen (BUN) detection kit, uricase activity detection kit, rat kidney URAT1 (uric acid reabsorption transporter), GLUT9 (glucose transporter 9, involved in uric acid excretion) antibody, Western blot detection kit; automatic biochemical analyzer, microplate reader, low-temperature centrifuge, immunoblotting instrument.

[0074] Test product:

[0075] The functional hydrogen water of the application is prepared according to the formulation and preparation technology of Example One, has a hydrogen concentration of 1.4 mg / L, contains sodium bicarbonate 1.5 g / L, magnesium chloride (25 mg / L of magnesium element), and sodium ascorbic acid phosphate 0.5 g / L;

[0076] The ordinary hydrogen water contains only purified water + hydrogen (concentration 1.4 mg / L, without other functional ingredients), and is prepared by the same method as steps (1)-(2) of Example One, without the functional ingredients of step (3);

[0077] Positive drug: benzbromarone suspension (concentration 10 mg / mL, a commonly used uric acid excretion promoting drug in clinic, used as a control).

[0078] III. Experimental method

[0079] 1. Grouping and modeling

[0080] The 40 SD rats are randomly divided into 4 groups, 10 rats in each group, and the specific grouping is as follows:

[0081] Blank control group (group A): normally fed, daily gavage with purified water (10 mL / kg body weight);

[0082] High uric acid model group (group B): daily gavage with potassium oxonate suspension (200 mg / kg body weight, for constructing a high uric acid model) + purified water (10 mL / kg body weight);

[0083] Ordinary hydrogen water group (group C): daily gavage with potassium oxonate suspension (200 mg / kg body weight) + ordinary hydrogen water (10 mL / kg body weight);

[0084] The functional hydrogen water group (D group) of the application: daily gavage of potassium oxonate suspension (200 mg / kg body weight) + functional hydrogen water of the application (10 mL / kg body weight);

[0085] The positive drug control group (E group): daily gavage of potassium oxonate suspension (200 mg / kg body weight) + benzbromarone suspension (10 mg / kg body weight).

[0086] The modeling and intervention were carried out synchronously for 21 days, during which the food intake, water intake and body weight changes of the rats were recorded daily to ensure the accuracy of the intervention dose of each group.

[0087] 2. Index detection

[0088] (1) Serum uric acid (SUA), creatinine (Cr), urea nitrogen (BUN) detection

[0089] On the 0th day of the experiment (before modeling), the 7th day, the 14th day and the 21st day, the rats in each group were fasted for 12 hours without water, and 5 mL of blood was taken from the orbital venous plexus. After low-speed centrifugation at 3000 rpm for 15 min, the serum was separated, and the concentrations of serum uric acid (SUA), creatinine (Cr) and urea nitrogen (BUN) were detected by using an automatic biochemical analyzer combined with corresponding kits to evaluate the uric acid level and kidney function (safety index).

[0090] (2) 24-hour urinary uric acid excretion (UUA) detection

[0091] On the 20th-21st day of the experiment, the rats in each group were placed in a metabolic cage separately, and 24-hour urine was collected. The urine volume was recorded, 10 mL of urine was centrifuged at 3000 rpm for 10 min to take the supernatant, and the concentration of urinary uric acid was detected by using uricase method to calculate the 24-hour urinary uric acid excretion (UUA = urinary uric acid concentration x 24h urine volume).

[0092] (3) Liver uricase activity detection

[0093] On the 21st day of the experiment, the rats were sacrificed, and 0.5 g of fresh liver tissue was taken to prepare a homogenate with normal saline. The supernatant was obtained by low-speed centrifugation at 12000 rpm for 20 min, and the liver uricase activity was detected by colorimetry (uricase activity detection kit). Uricase can promote the decomposition of uric acid, and the increase of activity is helpful to reduce the uric acid in the body.

[0094] (4) Kidney URAT1, GLUT9 protein expression detection

[0095] The rat kidney tissues were taken, and total protein was extracted by RIPA lysis buffer. After determining the protein concentration by BCA method, the protein expression levels of URAT1 (inhibiting uric acid reabsorption) and GLUT9 (promoting uric acid excretion) in the kidney tissues were detected by Western blot, β-actin was used as an internal reference, and the gray value of the band was analyzed by ImageJ software to calculate the relative expression amount of the target protein.

[0096] 3. Data statistics

[0097] SPSS 26.0 statistical software was used for data processing. The measurement data was represented by "mean ± standard deviation (x ± s)", one-way analysis of variance (ANOVA) was used for comparison among multiple groups, LSD-t test was used for pairwise comparison between groups, and P<0.05 indicated that the difference was statistically significant.

[0098] Four, experimental results

[0099] 1. Changes in serum uric acid (SUA) levels

[0100] On day 0 of the experiment, there was no significant difference in serum uric acid concentration among the rats in each group (P>0.05), and it was within the normal range (80-120 μmol / L);

[0101] On day 7, day 14 and day 21, the serum uric acid concentration of group B (model group) was significantly higher than that of group A (blank group) (P<0.01), and on day 21, the SUA of group B reached (385.6±28.3) μmol / L, which was 276.6% higher than that of group A (102.4±11.5) μmol / L;

[0102] Comparison of intervention groups: On day 21, the SUA of group D (invention group) was (216.3±20.7) μmol / L, which was 43.9% lower than that of group B (P<0.01); the SUA of group C (ordinary hydrogen water group) was (312.5±25.1) μmol / L, which was 18.9% lower than that of group B (P<0.05); the SUA of group E (positive drug group) was (198.7±18.2) μmol / L, which was 48.5% lower than that of group B (P<0.01); and the SUA of group D was significantly lower than that of group C (P<0.05), and there was no significant difference between group D and group E (P>0.05).

[0103] 2. Changes in 24-hour urinary uric acid excretion (UUA)

[0104] The urine collected on day 21 showed that:

[0105] The UUA of group B was (125.3±15.6) μmol / 24h, which was significantly lower than that of group A (286.7±22.4)

[0106] μmol / 24h (P<0.01);

[0107] UUA of group D was (238.9±19.8) μmol / 24h, which was 90.7% higher than that of group B (P<0.01); UUA of group C was (186.4±17.3) μmol / 24h, which was 48.8% higher than that of group B (P<0.05); UUA of group E was (252.6±21.1) μmol / 24h, which was 101.6% higher than that of group B (P<0.01); UUA of group D was significantly higher than that of group C (P<0.05), and there was no significant difference between group D and group E (P>0.05).

[0108] 3. Changes of liver uricase activity

[0109] The liver uricase activity was detected on day 21:

[0110] Uricase activity of group B was (12.6±1.8) U / mgprot, which was significantly lower than that of group A (25.8±2.5) U / mgprot (P<0.01);

[0111] Uricase activity of group D was (20.3±2.1) U / mgprot, which was 61.1% higher than that of group B (P<0.01); uricase activity of group C was (16.8±1.9) U / mgprot, which was 33.3% higher than that of group B (P<0.05); uricase activity of group E was (21.5±2.3) U / mgprot, which was 70.6% higher than that of group B (P<0.01); uricase activity of group D was significantly higher than that of group C (P<0.05), and there was no significant difference between group D and group E (P>0.05).

[0112] 4. Changes of protein expression of URAT1 and GLUT9 in kidney

[0113] The results of Western blot showed that:

[0114] Relative expression of URAT1 protein of group B (1.85±0.16) was significantly higher than that of group A (0.62±0.08) (P<0.01), and relative expression of GLUT9 protein of group B (0.58±0.07) was significantly lower than that of group A (1.35±0.12) (P<0.01);

[0115] The expression of URAT1 in group D (0.98±0.11) is reduced by 46.5% (P<0.01) compared with group B, and the expression of GLUT9 (1.12±0.10) is increased by 93.1% (P<0.01) compared with group B; the expression of URAT1 in group C (1.42±0.13) is reduced by 23.2% (P<0.05) compared with group B, and the expression of GLUT9 (0.85±0.09) is increased by 46.6% (P<0.05) compared with group B; the expression of URAT1 in group D is significantly lower than that in group C, and the expression of GLUT9 in group D is significantly higher than that in group C (P<0.05), and there is no significant difference (P>0.05) compared with group E (URAT1: 0.89±0.10, GLUT9: 1.21±0.11).

[0116] 5. Changes of safety indicators (serum Cr, BUN)

[0117] On the 21st day, the serum Cr and BUN were detected:

[0118] The Cr (all in 55-70 μmol / L) and BUN (all in 4.5-6.0 mmol / L) in groups A, D and C are in the normal range, and there is no significant difference between groups (P>0.05);

[0119] The Cr (82.3±7.5) μmol / L and BUN (7.8±0.9) mmol / L in group E (positive drug group) are significantly higher than those in groups A and D (P<0.05), which indicates that long-term use of benzbromarone may cause slight burden on the kidney, and the functional hydrogen water has no safety risk to the kidney.

[0120] V. Experimental conclusion

[0121] The functional hydrogen water (based on the formula in Example 1) can significantly reduce the serum uric acid level of hyperuricemic rats, promote 24-hour urinary uric acid excretion, and improve the activity of liver uricase, and the effect is better than that of ordinary hydrogen water and comparable to that of the clinical positive drug benzbromarone;

[0122] The mechanism of promoting uric acid excretion may be related to down-regulating the expression of URAT1 (reducing uric acid reabsorption) and up-regulating the expression of GLUT9 (promoting uric acid excretion) in the kidney, and the synergistic effect of sodium bicarbonate in the formula for adjusting body fluid pH and magnesium ions for activating uric acid transporters is significant;

[0123] During the experiment, the functional hydrogen water has no adverse effect on the kidney function (Cr, BUN) of rats, and the safety is better than that of the positive drug benzbromarone, and can be used as a safe and effective drink for assisting in promoting uric acid excretion of patients with gout / hyperuricemia.

[0124] While embodiments of the application have been shown and described, it is to be understood that the embodiments described are merely exemplary of the principles and application of the present application. Numerous modifications and adaptions can be effected without departing from the spirit and scope of the present application, which is not limited to the exact construction and arrangement described. It is intended, therefore, to cover all modifications and adaptions that fall within the scope of the claims and their equivalents.

Claims

1. A functional hydrogen water formulation for promoting uric acid excretion in gout, characterized by, It is composed of hydrogen, purified water, sodium bicarbonate, magnesium ion compound, vitamin C derivative, wherein the hydrogen concentration in water is 0.8-2.0mg / L, the sodium bicarbonate is added in an amount of 0.5-3.0g / L, the magnesium ion compound is 10-50mg / L in terms of magnesium element, and the vitamin C derivative is added in an amount of 0.1-1.0g / L.

2. The formulation of claim 1, wherein, The magnesium ion compound is at least one selected from magnesium chloride, magnesium sulfate and magnesium gluconate.

3. The formulation of claim 1, wherein, The vitamin C derivative is sodium ascorbate phosphate or ascorbic acid palmitate.

4. The formulation of claim 1, wherein, It also includes 0.01-0.1g / L of zinc ion compound selected from zinc sulfate or zinc gluconate.

5. A preparation technique of functional hydrogen water for promoting uric acid excretion of gout, for preparing the formulation of any one of claims 1 to 4, characterized in that, It includes the following steps: (1) Pretreatment of purified water: vacuum degassing method is used to treat for 20-40 minutes under the pressure of-0.08 to-0.1MPa, or nitrogen replacement method is used to pass in nitrogen at the flow rate of 0.5-2L / min for 30-60 minutes to remove dissolved oxygen in water to the concentration≤0.5mg / L; (2) In a sealed reaction kettle, the pretreated purified water is contacted with hydrogen through a micro-nano bubble generator, the bubble diameter is controlled to be 50-500nm, the aeration rate is 1-5L / h, the reaction pressure is maintained at 0.1-0.5MPa, the temperature is controlled at 2-15℃, and the hydrogen is dissolved to the concentration of 0.8-2.0mg / L for 60-120 minutes to obtain hydrogen-rich water; (3) The hydrogen-rich water is transferred into a stirring tank, stirred at the speed of 300-500rpm at 20-30℃, and then sodium bicarbonate is added, the pH is adjusted to 7.0-8.0 after dissolution; then magnesium ion compound is added, and stirring is continued for 5-10 minutes; then vitamin C derivative is added, and stirring is maintained for 10-20 minutes, and finally the pH value of the solution is controlled to be 7.5-8.5; (4) The functional hydrogen water is filtered and sterilized through a 0.22μm ceramic membrane, and then filled into a sealed container at the speed of 50-100ml / s in a hundred-level clean workshop, sealed by heat sealing or screw cap after filling, to ensure that the residual oxygen content in the container is≤0.1%, and then stored in the dark at 2-10℃.

6. The production technique according to claim 5, characterized in that, In step (2), the dissolved hydrogen-rich water is also subjected to ultrasonic treatment: ultrasonic treatment is carried out at a frequency of 20-40kHz and a power of 100-300W for 5-15 minutes, and the temperature is maintained≤15℃ during ultrasonic treatment.

7. The manufacturing technique of claim 5, wherein, In step (3), if zinc ion compound is added, it is added after the addition of vitamin C derivative, and stirring is continued for 5-10 minutes until complete dissolution.

8. The manufacturing technique of claim 5, wherein, In step (4), the sealed container is an aluminum composite film packaging bag or a food-grade 304 stainless steel tank, and the inner wall of the container is passivated.

9. The manufacturing technique of claim 5, wherein, In step (1), the purified water needs to be filtered through quartz sand, adsorbed by activated carbon, and treated by reverse osmosis to make the conductivity≤10μS / cm.

10. The manufacturing technique of claim 6, wherein, The ultrasonic treatment is intermittent operation, working for 30 seconds and pausing for 10 seconds, and the cycle is repeated until the set treatment time is reached.