A method for enhancing the inhibitory activity of sophora flavescens against xanthine oxidase by acid heat treatment

Abstract

The application discloses a method for enhancing the xanthine oxidase inhibitory activity of sophora fruit by acid heat treatment, and belongs to the technical field of food deep processing. The method specifically comprises the following steps: cleaning fresh sophora fruit, adding an organic acid solution according to a certain material-liquid ratio, and soaking at room temperature, draining water from the sophora fruit after the soaking treatment, and performing hot air drying or high-temperature stir-frying drying, and the sophora fruit is obtained. The polyphenol components in the sophora fruit product prepared by the method change significantly, and the sophora fruit product has better in-vivo xanthine oxidase inhibitory activity than before the treatment; and the method is simple in operation, and can be easily used for industrialized production of sophora fruit products with xanthine oxidase inhibitory activity.

Description

Technical Field

[0001] This invention relates to a method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, belonging to the field of food deep processing technology. Background Technology

[0002] Uric acid is produced by the metabolism of purines in the human body, including adenine nucleotides, hypoxanthine nucleotides, and guanine nucleotides. Purine nucleotides are first metabolized into xanthine, which is then catalyzed by xanthine oxidase to produce uric acid. Disorders of purine metabolism can lead to hyperuricemia, which is diagnosed when serum uric acid levels are >7 mg / dL in men and >5.7 mg / dL in women. Due to the intake of high-purine foods and unhealthy dietary habits, the global population with hyperuricemia is showing a year-on-year upward trend, becoming a significant threat to human health. Xanthine oxidase is a key enzyme controlling uric acid production in the human body; inhibiting xanthine oxidase activity is an important pathway for regulating uric acid levels in the body.

[0003] Currently available xanthine oxidase inhibitors include allopurinol and febuxostat, which all work by inhibiting xanthine oxidase activity and blocking uric acid production to lower uric acid levels. However, most patients with hyperuricemia have serum uric acid levels near the borderline and do not require medication. Furthermore, these medications have shown various side effects during long-term clinical use. Therefore, developing naturally derived xanthine oxidase inhibitors has become a hot topic for researchers and manufacturers.

[0004] Sophora japonica buds (Flos Sophorae Immaturus), native to my country, are also known as white sophora japonica, sophora flower rice, or sophora seeds. They are made from the dried flower buds of the Sophora japonica L., a legume. As a plant used for both food and medicine, Sophora japonica buds were already being used by the ancients in the Ming Dynasty to treat aphonia, stop bleeding, and sore throat. Modern research shows that Sophora japonica bud tea can be consumed as a health food to lower blood sugar, blood lipids, and blood pressure, and also has antioxidant and immune-boosting effects. Sophora japonica buds are rich in flavonoids (dry basis content can reach over 20%), mainly including rutin, quercetin, kaempferol-3-O-rutin, and narcissin. As previously studied, Chinese patent CN115211481A discloses a Sophora japonica bud tea and its preparation method. This method uses ultrasonic treatment combined with high-temperature drying to increase the flavonoids in Sophora japonica buds, thereby showing a good effect in inhibiting xanthine oxidase activity in vitro. However, the product obtained by this method still has relatively low inhibitory activity against xanthine oxidase in vivo. Summary of the Invention

[0005] To address the shortcomings and deficiencies of existing technologies, this invention provides a method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment. This method can effectively solve problems such as reduced xanthine oxidase inhibitory activity in existing Sophora japonica bud processing products and cumbersome processing procedures.

[0006] The purpose of this invention is to provide a method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment. The method includes: washing fresh Sophora japonica buds, adding an organic acid solution at a certain material-to-liquid ratio and soaking at room temperature, draining the soaked Sophora japonica buds, and then drying them with hot air or by high-temperature stir-frying.

[0007] In one embodiment, the method further includes pulverizing and extracting dried Sophora japonica buds to obtain Sophora japonica bud extract.

[0008] In one embodiment, the material-to-liquid ratio refers to the ratio formed by adding fresh Sophora japonica flowers to an organic acid solution, and the material-to-liquid ratio is 1:0.5 to 1.5.

[0009] In one embodiment, the organic acid solution includes one or more of malic acid, lactic acid, and oxalic acid; preferably oxalic acid.

[0010] In one embodiment, the organic acid solution has a mass concentration of 2% to 10%.

[0011] In one embodiment, the soaking time is 60 min to 120 min.

[0012] In one embodiment, the hot air drying temperature is 160℃~200℃, and the drying time is 30min~90min.

[0013] In one embodiment, the high-temperature stir-frying temperature is 180℃~220℃, the time is 20min~60min, and the stir-frying speed is 30r / min~120r / min.

[0014] Another object of the present invention is to provide a Sophora japonica flower obtained after processing by the above method.

[0015] A third objective of this invention is to provide the application of the aforementioned Sophora japonica flowers in the preparation of food and health products.

[0016] The beneficial effects of this invention are:

[0017] This invention uses fresh Sophora japonica buds as raw material. The buds are soaked in an organic acid solution, allowing the acid radicals to fully infiltrate the cell walls and interact with polyphenols. This is followed by heat treatment such as hot air or high-temperature roasting, which significantly alters the composition of the phenolic compounds in the buds at 160℃–220℃ for 20–90 minutes (rutin, hyperoside, and quercetin are converted to quercetin; kaempferol-3-O-rutinoside is converted to kaempferol; and narcisin is converted to isorhamnetin). Furthermore, the acid-heat treatment significantly enhances the xanthine oxidase inhibitory activity of the polyphenols in the Sophora japonica buds, resulting in better biological activity. This method is simple to operate and suitable for industrial production. Attached Figure Description

[0018] Figure 1 High performance liquid chromatography and liquid mass spectrometry ion fragmentation diagrams of polyphenol changes in the Sophora japonica products of Example 6 and Comparative Example 1. Detailed Implementation

[0019] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention.

[0020] Example 1

[0021] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0022] (1) Clean the fresh Sophora japonica flowers, add a 5% malic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 60 minutes;

[0023] (2) Drain the water from the soaked Sophora japonica flowers in step (1), dry them with hot air at 180℃ for 30 minutes.

[0024] Example 2

[0025] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0026] (1) Clean the fresh Sophora japonica flowers, add 5% lactic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 60 minutes;

[0027] (2) Drain the water from the soaked Sophora japonica flowers in step (1), dry them with hot air at 180℃ for 30 minutes.

[0028] Example 3

[0029] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0030] (1) Clean the fresh Sophora japonica flowers, add 5% oxalic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 60 minutes;

[0031] (2) Drain the water from the soaked Sophora japonica flowers in step (1), dry them with hot air at 180℃ for 30 minutes.

[0032] Example 4

[0033] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0034] (1) Clean the fresh Sophora japonica flowers, add a 10% malic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes.

[0035] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature for 30 minutes at 200℃ and 60r / min.

[0036] Example 5

[0037] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0038] (1) Clean the fresh Sophora japonica flowers, add 2% oxalic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 60 minutes;

[0039] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature for 30 minutes at 180℃ and 60r / min.

[0040] Example 6

[0041] A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, comprising the following steps:

[0042] (1) Clean the fresh Sophora japonica flowers, add 10% oxalic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0043] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature, adjust the temperature to 180℃, stir-fry for 60 minutes, and rotate at 60 r / min.

[0044] Comparative Example 1

[0045] (1) Clean the fresh Sophora japonica flowers, add distilled water at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0046] (2) Drain the water from the soaked Sophora japonica flowers in step (1) and let them air dry for 1-2 days, turning them over every 30 minutes during the drying process.

[0047] Comparative Example 2

[0048] (1) Clean the fresh Sophora japonica flowers, add distilled water at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0049] (2) Drain the water from the soaked Sophora japonica flowers in step (1), dry them with hot air at 60°C for 300 minutes.

[0050] Comparative Example 3

[0051] (1) Clean the fresh Sophora japonica flowers, add distilled water at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0052] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature, adjust the temperature to 180℃, stir-fry for 60 minutes, and rotate at 60 r / min.

[0053] Comparative Example 4

[0054] (1) Clean the fresh Sophora japonica flowers, add 10% acetic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0055] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature, adjust the temperature to 180℃, stir-fry for 60 minutes, and rotate at 60 r / min.

[0056] Comparative Example 5

[0057] (1) Clean the fresh Sophora japonica flowers, add 10% citric acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0058] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature, adjust the temperature to 180℃, stir-fry for 60 minutes, and rotate at 60 r / min.

[0059] Comparative Example 6

[0060] (1) Clean the fresh Sophora japonica flowers, add 10% oxalic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0061] (2) Drain the water from the soaked Sophora japonica flowers in step (1), stir-fry them at high temperature, adjust the temperature to 100℃, stir-fry for 60 minutes, and rotate at 60 r / min.

[0062] Comparative Example 7

[0063] (1) Clean the fresh Sophora japonica flowers, add 10% oxalic acid solution at a material-to-liquid ratio of 1:1 and soak at room temperature for 120 minutes;

[0064] (2) Drain the water from the soaked Sophora japonica flowers in step (1), dry them with hot air at 60°C for 300 minutes.

[0065] Comparative Example 8

[0066] (1) Wash the fresh Sophora japonica flowers clean, add distilled water solution according to the material-to-liquid ratio of 1:1, place in a hydrothermal processor, adjust the temperature to 180℃, and time to 60min.

[0067] (2) Drain the water from the Sophora japonica flowers after hydrothermal treatment in step (1), and dry them with hot air at 60°C for 300 minutes.

[0068] Performance testing

[0069] The Sophora japonica buds obtained in Examples 1-6 and Comparative Examples 1-8 were pulverized using a pulverizer and passed through an 80-mesh sieve to obtain Sophora japonica bud powder. The obtained Sophora japonica bud powder was added to a 70% ethanol solution at a material-to-liquid ratio of 1:400 (m / v), and extracted for 40 min on a magnetic stirrer at 250 r / min at an extraction temperature of 25℃. The extraction was repeated three times. The extracts were combined and rotary evaporated to near dryness. The solution was dissolved in 70% ethanol and diluted to 500 mL to obtain the Sophora japonica bud extract. The content of the main polyphenols in the Sophora japonica buds was determined by high performance liquid chromatography, and the results are shown in Table 1.

[0070] Table 1 shows the changes in the content of rutin, hyperoside, quercetin, quercetin, kaempferol-3-O-rutin, kaempferol, narcissin, and isorhamnetin in the examples and comparative examples.

[0071]

[0072]

[0073] Sophora japonica extract was purified using D101 macroporous resin. The extract was loaded into a column (inner diameter × L = 2 cm × 30 cm) at a rate of 1.5 mL / min. Sugars and other components were then washed with distilled water until the eluent was colorless. Subsequently, the polyphenolic extract of Sophora japonica was desorbed from the macroporous resin with 70% ethanol at a rate of 1.5 mL / min. The eluent was collected and rotary evaporated at 50 °C, followed by freeze-drying to obtain the purified polyphenolic extract of Sophora japonica.

[0074] A mouse model of hyperuricemia was established by gavage administration of hypoxanthine (500 mg / kg bw) and intraperitoneal injection of potassium oxonate (200 mg / kg bw). The mice were then administered purified Sophora japonica polyphenol extract (200 mg / kg bw) by gavage. The control group received 0.5% sodium carboxymethyl cellulose by gavage, while the positive control group received allopurinol (10 mg / kg bw) by gavage. Serum and liver xanthine oxidase activity were measured using a kit. The results are shown in Table 2.

[0075] Table 2. In vivo xanthine oxidase inhibitory activity of the examples and comparative examples

[0076]

[0077]

[0078] The results in Tables 1 and 2 show that:

[0079] (1) Compared with Examples 1-6, the Sophora japonica flowers in Comparative Examples 1-2 were not soaked in organic acid and subjected to high-temperature heat treatment. Their serum and liver xanthine oxidase activities were significantly lower than those in Examples 1-6, indicating that acid heat treatment can significantly enhance the xanthine inhibitory activity of Sophora japonica flowers.

[0080] (2) Compared with Examples 1-6, in Comparative Example 3, the Sophora japonica flowers were not soaked in acid, and the degradation rates of rutin, hyperoside, quercetin, kaempferol-3-O-rutin, and narcissin were lower than those in Examples 1-6. The increase rates of quercetin, kaempferol, and isorhamnetin were also lower than those in Examples 1-6. The inhibitory activities of xanthine oxidase in serum and liver were significantly lower than those in Examples 1-6. However, compared with Comparative Examples 1-2, the degradation rate and increase rate of the main polyphenols, as well as the inhibitory activities of xanthine oxidase in serum and liver, of the product of Comparative Example 3 after high-temperature heat treatment were significantly higher than those in Comparative Examples 1-2. This indicates that acid treatment can synergistically promote the degradation and transformation of Sophora japonica polyphenols with heat treatment, and further enhance the xanthine oxidase inhibitory activity of Sophora japonica polyphenols.

[0081] (3) The serum and liver xanthine oxidase inhibitory activities of Examples 1-3 were, in descending order, Example 3 > Example 2 > Example 1, indicating that oxalic acid treatment had the most significant effect on enhancing xanthine oxidase inhibitory activity, followed by lactic acid and malic acid.

[0082] (4) Compared with Example 6 and Comparative Example 3, Comparative Examples 4-5 were soaked in acetic acid or citric acid and then subjected to high-temperature heat treatment. Their main polyphenol degradation rate and conversion rate, serum and liver xanthine oxidase inhibitory activity were not significantly different from those of Comparative Example 3, and were significantly lower than those of Example 6. This indicates that acetic acid or citric acid cannot synergistically enhance the xanthine oxidase inhibitory activity of Sophora japonica polyphenols with heat treatment.

[0083] (5) Compared with Example 6, Comparative Example 6 used oxalic acid soaking combined with 100°C heat treatment, and Comparative Example 7 used oxalic acid soaking combined with 60°C heat treatment. Neither of the two methods was subjected to high temperature treatment of 160°C to 220°C. The degradation rate and conversion rate of the main polyphenols, as well as the inhibitory activity of xanthine oxidase in serum and liver, were significantly lower than those of Example 6. This indicates that organic acid combined with ordinary heat treatment, without high temperature heat treatment, cannot achieve the purpose of enhancing the inhibitory activity of xanthine oxidase in Sophora japonica polyphenols.

[0084] (6) Compared with Example 6, Comparative Example 8 was treated with hydrothermal treatment. The heating temperature and time were the same as those in Example 6, but its main polyphenol degradation rate and conversion rate, as well as the inhibitory activity of xanthine oxidase in serum and liver, were significantly lower than those in Example 6, indicating that acid-heat combined treatment is more advantageous than hydrothermal treatment.

[0085] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for enhancing the inhibitory activity of Sophora japonica buds against xanthine oxidase through acid heat treatment, characterized in that, The method includes: washing fresh sophora japonica flowers clean, adding organic acid solution according to a certain material-liquid ratio and soaking at room temperature, draining the water after soaking, and then drying with hot air or high-temperature stir-frying. The material-to-liquid ratio is 1:0.5~1.5; the mass concentration of the organic acid solution is 2%~10%; The organic acid solution includes one or more of malic acid, lactic acid, and oxalic acid; The hot air drying temperature is 160℃~200℃, and the drying time is 30 min~90 min; Alternatively, the high-temperature stir-frying temperature is 180℃~220℃, the time is 20 min~60 min, and the stir-frying speed is 30 r / min~120 r / min.

2. The method according to claim 1, characterized in that, The method also includes pulverizing and extracting dried Sophora japonica buds to obtain Sophora japonica bud extract.

3. The method according to claim 1, characterized in that, The soaking time is 60 min to 120 min.

4. Sophora japonica buds or Sophora japonica bud extract obtained after treatment by the method according to any one of claims 1 to 3.

5. The application of the Sophora japonica buds or Sophora japonica bud extract as described in claim 4 in the preparation of food and health products.

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