Quinoa alkali extract with anti-fatigue activity and preparation method thereof

Abstract

The invention discloses a chenopodium quinoa alkali extract with anti-fatigue activity and a preparation method of the chenopodium quinoa alkali extract. According to the method, chenopodium quinoa is used as a raw material, and a food-grade calcium oxide mixed solution medium is adopted for extraction. Chenopodium quinoa willd is rinsed, dried, crushed and sieved, a food-grade calcium oxide mixed solution is extracted and centrifuged, and supernate is diluted and spray-dried to obtain the quinoa willd alkali extract. The method is simple to operate, low in cost and high in extraction rate, secondary chemical components of quinoa starch and crude fibers can be removed to a large extent, the most important nutritional components with health-care functions of quinoa are reserved, and the biological activity of the nutritional components of the extract is kept. The quinoa alkali extract disclosed by the invention can remarkably prolong the swimming time of mice, reduce the content of blood lactic acid and urea nitrogen and increase the content of hepatic glycogen at the same time, and shows a relatively good anti-fatigue effect. The obtained quinoa alkali extract can be used for making anti-fatigue functional foods such as compressed biscuits and tablet candies.

Description

TECHNICAL FIELD

[0001] The present application relates to a quinoa alkaline extract with anti-fatigue activity and a preparation method thereof, belonging to the technical field of natural product extraction and functional food. BACKGROUND

[0002] Fatigue is a common physiological and psychological state characterized by physical and mental decline, often accompanied by symptoms such as lack of concentration, slow reaction, and muscle weakness. It is widespread in modern society, especially after high-intensity work, study, or exercise. Fatigue is related to various factors, including prolonged physical or mental work, sleep deprivation, malnutrition, and psychological stress. In recent years, research has found that fatigue is closely related to the immune system, neuroendocrine system, oxidative stress, and changes in gut flora. Chronic fatigue not only affects quality of life, but also can lead to various chronic diseases such as cardiovascular disease, immune dysfunction, and nervous system disease (Li Ruili, Hu Di, Cui Yilong, et al. Neuroinflammatory mechanisms of chronic fatigue syndrome [J]. International Journal of Psychiatry, 2020, 47(06): 1113-1117). Currently, anti-fatigue products on the market are mainly chemical synthetic drugs, which may have side effects with long-term use. Therefore, developing natural, safe, and efficient anti-fatigue active ingredients has become a research hotspot.

[0003] Quinoa (Chenopodium quinoa Wild.) is a pseudo-cereal originally from the Andes region of South America. It is considered an important crop for global food security due to its outstanding nutritional value and adaptability (Zhang Shuwei, Zong Yingjie, Huang Linli, et al. Analysis of research progress and hotspots of quinoa in China and abroad in the past decade [J]. China Agricultural Bulletin, 2024, 40(05): 145-152). Quinoa seeds are rich in high-quality protein, dietary fiber, vitamins, minerals, and various bioactive components such as polyphenols, flavonoids, and saponins. The protein content is about 13.1% to 16.7%, significantly higher than traditional cereals such as wheat, corn, and rice, and the amino acid composition is balanced, especially the high content of lysine, which can meet the human body's demand for essential amino acids (Vilcacundo R, Hernandez-Ledesma B. Nutritional and biological value of quinoa (Chenopodium quinoa Willd.) [J]. Current Opinion in Food Science, 2016, 14: 1-6).

[0004] Quinoa, as a functional food with anti-fatigue potential, has a broad development prospect. Quinoa is rich in high-quality protein, dietary fiber, vitamins, minerals, and various antioxidant components such as polyphenols and flavonoids. These components have significant antioxidant activity, can scavenge free radicals in the body, reduce oxidative stress, and thus play an anti-fatigue role. For example, high-quality protein and balanced amino acid composition, especially the high content of branched-chain amino acids, can effectively promote muscle repair after exercise; including arabinoxylan with immune regulation function and rhamnose galacturonic acid glycan with anti-fatigue properties; in addition, saponins in quinoa have anti-inflammatory and antioxidant properties, which can enhance the body's endurance and recovery ability (Hu Y C, Zhao G, Qin P Y, et al. Research progress of active components in quinoa [J]. Crop Science, 2018, 44(11): 1579-1591).

[0005] Yuan J J, Jiang Y R, Sun X T, et al. Optimization of polysaccharide extraction process from quinoa by response surface methodology and its variety differences [J]. Food Science and Technology, 2016, 41(01): 154-159) determined the optimal water extraction process of quinoa seed polysaccharides through single factor and response surface experiments: under the conditions of extraction temperature 91℃, time 1.5h, and solid-liquid ratio 1:34, the theoretical extraction rate of crude polysaccharides reached 15.81%. Studies have shown that the degree of influence of each factor is in the order of solid-liquid ratio > extraction temperature > extraction time, which provides a basis for the industrial extraction of quinoa polysaccharides. However, this method still has limitations: single water extraction method has insufficient extraction rate of combined polysaccharides and long-time heating may cause damage to heat-sensitive active ingredients, especially for active polysaccharide components combined with protein in quinoa, conventional water extraction method is difficult to effectively release. Therefore, developing an efficient, stable and anti-fatigue active extraction method for quinoa has become a technical problem to be solved in the field.

[0006] Bai H J, Pang W Q, Zhang Z H, et al. Fermentation process of quinoa-blue huckleberry compound juice and antioxidant and anti-fatigue effects of its supernatant [J]. Food and Machinery, 2024, 40(07): 148-154) further explored the anti-fatigue efficacy of quinoa through fermentation and other technical means. For example, some studies found that the fermentation product of quinoa-blue huckleberry compound juice by yeast and lactic acid bacteria had significant antioxidant and anti-fatigue effects, and could prolong the exhaustive swimming time of mice under load. This fermentation process not only improves the functionality of quinoa, but also provides technical support for the development of new anti-fatigue foods. However, this fermentation technology needs to strictly control pH and temperature, and the fermentation period is as long as 48h, which significantly increases the cost of industrial production and makes it difficult to achieve large-scale application.

[0007] Zhou Yali et al. (Zhou Yali, Jiang Quan. Study on the Anti-fatigue Effect of Quinoa Protein Peptide QPP-I in Mice [J]. Food Science and Technology, 2024, 49(07): 196-204) used quinoa protein as raw material to prepare quinoa protein peptide by enzymatic hydrolysis and membrane ultrafiltration separation method and conducted anti-fatigue experiment in mice. The results showed that the swimming time of mice ingesting quinoa protein peptide was significantly prolonged, the levels of lactic acid and urea nitrogen in blood were reduced, and the contents of liver and muscle glycogen were increased. This study provides certain theoretical and technical support for the development of high-value natural quinoa antioxidant peptide products.

[0008] With the increasing demand for healthy food among consumers, quinoa has a broad market application prospect in the field of anti-fatigue. Its extracts and fermented products can be widely used in food, health care and pharmaceutical industries. CN106107463A patent discloses a super quinoa anti-fatigue milk tea making process. This patent relates to a super quinoa anti-fatigue milk tea making process, which includes the steps of raw material processing, soaking, grinding, filtering, homogenizing and concentrating, freeze-drying, crushing and sieving, and mixing and stirring. The final product has the effects of tonifying liver and kidney, invigorating spleen and stomach, nourishing heart and calming mind, etc., and can relieve physical fatigue, improve anti-hypoxia tolerance, and enhance immune function. This patent effectively solves the problems of low active ingredient content, unclear efficacy, and single application form in existing quinoa anti-fatigue products, and provides a reliable technical solution for the development of a new generation of natural anti-fatigue functional food.

[0009] In summary, quinoa has great development potential and is expected to play an important role in the field of healthy food and health care products due to its rich nutritional ingredients, significant anti-fatigue effects, and broad market demand. Through retrieval, no patent disclosure literature related to this patent application has been found. SUMMARY

[0010] The purpose of the present application is to extract active ingredients from quinoa using an alkali method and provide its application in anti-fatigue activity. The specific preparation steps of the quinoa alkali extract of the present application are as follows:

[0011] (1) Wash the quinoa raw material with deionized water, drain the water, and then place it in an oven at 60℃ for drying. After crushing and passing through a 50 mesh sieve, quinoa powder is obtained;

[0012] (2) Put the quinoa powder obtained in step (1) into an extraction tank, mix it with deionized water at a ratio of 1:10 to 1:14, add 0.2% food-grade calcium oxide mixed solution to adjust the pH of the extraction liquid to 8.0 to 9.0, set the extraction temperature to 30℃ to 50℃, and extract for 1.5 to 2.5 hours under these conditions;

[0013] (3) The extraction solution obtained in step (2) is transferred to a centrifuge for solid-liquid separation, and the centrifuge is set to rotate at 6000 r / min for 10 min to obtain a precipitate and a supernatant;

[0014] (4) The supernatant obtained in step (3) is diluted to twice the original volume and then subjected to spray drying under the conditions of an inlet temperature of 128 ℃, an outlet temperature of 60-65 ℃, a fan speed of 97%, a peristaltic pump speed of 25%, and a needle interval of 5 s to obtain a quinoa alkaline extract.

[0015] The advantages and positive effects of the present application are:

[0016] (1) The present application can maximize the removal of quinoa starch and crude fiber secondary chemical components, and retain the most important nutritional components with health care functions, including soluble quinoa protein, quinoa polysaccharide, quinoa total saponins, quinoa total flavonoids, quinoa total polyphenols, vitamins, and various mineral nutrients.

[0017] (2) The main active ingredients in the quinoa alkaline extract extracted by the present application can be greatly concentrated and enriched, and the solubility is also significantly improved. In the mouse weight-loaded swimming experiment, the swimming time of the mice is prolonged by 26.98%-109.26% compared with the blank; the blood lactic acid (LA) content is reduced by 25.08%-44.96% compared with the blank group; the urea nitrogen (BUN) content is reduced by 14.51%-37.09% compared with the blank group; and the liver glycogen content is increased by 28.29%-42.11% compared with the blank group.

[0018] (3) The traditional alkaline extraction method has a high extraction temperature or pH value, which not only easily denatures the protein and affects the functional properties of the product, but also increases the viscosity of the crude protein mother liquor, making separation difficult and increasing losses. When processed at high temperatures, the protein will undergo chemical changes such as racemization, hydrolysis, desulfurization, and deamidation, most of which are irreversible. In contrast, the present application has the advantages of short extraction time, mild conditions, low energy consumption, and suitability for large-scale industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 The raw material (quinoa raw material) of the anti-fatigue quinoa alkaline extract of the present application;

[0020] Figure 2 The anti-fatigue quinoa alkaline extract preparation process flow chart;

[0021] Figure 3 The anti-fatigue quinoa alkaline extract product (quinoa alkaline extract) of the present application;

[0022] Figure 4Effects of feeding different doses of quinoa alkaline extract on muscle tissue pathological morphology of exercise mice (A1 blank control group (before exercise), A2 blank control group (after exercise); B1 rhodioside control group (before exercise), B2 rhodioside control group (after exercise); C1 low-dose quinoa group (before exercise), C1 low-dose quinoa group (after exercise));

[0023] Figure 5 Effects of feeding different doses of quinoa alkaline extract on muscle tissue pathological morphology of exercise mice (A1 blank control group (before exercise), A2 blank control group (after exercise); B1 rhodioside control group (before exercise), B2 rhodioside control group (after exercise); D1 medium-dose quinoa group (before exercise), D2 medium-dose quinoa group (after exercise));

[0024] Figure 6 Effects of feeding different doses of quinoa alkaline extract on muscle tissue pathological morphology of exercise mice (A1 blank control group (before exercise), A2 blank control group (after exercise); B1 rhodioside control group (before exercise), B2 rhodioside control group (after exercise); E1 high-dose quinoa group (before exercise), E2 high-dose quinoa group (after exercise)). DETAILED DESCRIPTION

[0025] In order to better understand the present application, the present application will be further described in detail below in conjunction with examples, but the scope of protection claimed by the present application is not limited to the scope represented by the examples.

[0026] The following experimental steps are applied throughout the examples:

[0027] (1) The Kjeldahl method was used to determine the protein content of the quinoa alkaline extract.

[0028] (2) The phenol-sulfuric acid method was used to determine the polysaccharide content of the quinoa alkaline extract with glucose as the standard control.

[0029] (3) The vanillin-glacial acetic acid colorimetric method was used to determine the total saponin content of the quinoa alkaline extract with oleanolic acid as the standard control.

[0030] (4) The Folin phenol method was used to determine the total polyphenol content of the quinoa alkaline extract with gallic acid as the standard control.

[0031] (5) The sodium nitrite-aluminum nitrate method was used to determine the total flavonoid content of the quinoa alkaline extract with rutin as the standard control.

[0032] The present application will be further described below in conjunction with specific examples.

[0033] Example 1:

[0034] The quinoa raw material is washed with distilled water, drained and then placed in an oven at 60°C to dry. The dried material is ground and sieved through a 50 mesh sieve to obtain quinoa powder. 833g of the quinoa powder is added to 10L of deionized water in an extraction tank. The pH of the extraction solution is adjusted to 7.5 using a food-grade calcium oxide mixture. The extraction temperature is set to 40°C and the extraction is carried out for 2.0h. After the extraction is complete, the centrifuge speed is set to 6000r / min and the centrifugation is carried out for 10min. The supernatant is collected and diluted to 20L with deionized water. The solution is then spray dried under the following conditions: inlet temperature 128°C, outlet temperature 60°C-65°C, fan speed 97%, peristaltic pump speed 25%, and needle interval time 5s. Quinoa alkaline extract 183g is obtained. The polysaccharide content in the quinoa alkaline extract is 28.3%, the protein content is 40.2%, the total saponin content is 3.5%, the total polyphenol content is 3.6%, and the total flavonoid content is 1.5%.

[0035] Example 2:

[0036] The quinoa raw material is washed with distilled water, drained and then placed in an oven at 60°C to dry. The dried material is ground and sieved through a 50 mesh sieve to obtain quinoa powder. 833g of the quinoa powder is added to 10L of deionized water in an extraction tank. The pH of the extraction solution is adjusted to 8.0 using a food-grade calcium oxide mixture. The extraction temperature is set to 40°C and the extraction is carried out for 2.0h. After the extraction is complete, the centrifuge speed is set to 6000r / min and the centrifugation is carried out for 10min. The supernatant is collected and diluted to 20L with deionized water. The solution is then spray dried under the following conditions: inlet temperature 128°C, outlet temperature 60°C-65°C, fan speed 97%, peristaltic pump speed 25%, and needle interval time 5s. Quinoa alkaline extract 200g is obtained. The polysaccharide content in the quinoa alkaline extract is 30.50%, the protein content is 43.41%, the total saponin content is 4.1%, the total polyphenol content is 5.2%, and the total flavonoid content is 2.0%.

[0037] Example 3:

[0038] The quinoa raw material is washed with distilled water, drained and then placed in an oven at 60°C to dry. The dried material is ground and sieved through a 50 mesh sieve to obtain quinoa powder. 833g of the quinoa powder is added to 10L of deionized water in an extraction tank. The pH of the extraction solution is adjusted to 8.5 using a food-grade calcium oxide mixture. The extraction temperature is set to 40°C and the extraction is carried out for 2.0h. After the extraction is complete, the centrifuge speed is set to 6000r / min and the centrifugation is carried out for 10min. The supernatant is collected and diluted to 20L with deionized water. The solution is then spray dried under the following conditions: inlet temperature 128°C, outlet temperature 60°C-65°C, fan speed 97%, peristaltic pump speed 25%, and needle interval time 5s. Quinoa alkaline extract 220g is obtained. The polysaccharide content in the quinoa alkaline extract is 32.37%, the protein content is 45.27%, the total saponin content is 5.38%, the total polyphenol content is 6.0%, and the total flavonoid content is 2.4%.

[0039] Example 4:

[0040] Quinoa raw materials were washed with distilled water, drained, and dried in an oven at 60℃. The dried quinoa was then pulverized and passed through a 50-mesh sieve to obtain quinoa flour. 833g of quinoa flour was weighed and added to 10L of deionized water in an extraction tank. The pH of the extraction solution was adjusted to 9.0 with a food-grade calcium oxide mixture. The extraction temperature was set at 40℃, and extraction was carried out for 1.0 h under these conditions. After extraction, the mixture was centrifuged at 6000 rpm for 10 min. The supernatant was collected and diluted to 20L with deionized water, then spray-dried. The inlet temperature was set at 128℃, the outlet temperature at 60℃~65℃, the fan speed at 97%, the peristaltic pump speed at 30%, and the needle-passing interval at 5s. Under these conditions, 210g of quinoa alkaloid extract was obtained. The polysaccharide content, protein content, total saponin content, total polyphenol content, and total flavonoid content of the quinoa alkaloid extract were measured to be 31.87%, 44.5%, 4.72%, 5.4%, and 2.2%, respectively.

[0041] Example 5:

[0042] Quinoa raw materials were washed with distilled water, drained, and dried in an oven at 60℃. The dried quinoa was then pulverized and passed through a 50-mesh sieve to obtain quinoa flour. 1000g of quinoa flour was weighed and added to 10L of deionized water in an extraction tank. The pH of the extraction solution was adjusted to 8 with a food-grade calcium oxide mixture. The extraction temperature was set at 50℃, and extraction was carried out for 2.5 hours under these conditions. After extraction, the mixture was centrifuged at 6000 rpm for 10 minutes. The supernatant was collected and diluted to 20L with deionized water before spray drying. The inlet temperature was set at 128℃, the outlet temperature at 60℃~65℃, the fan speed at 97%, the peristaltic pump speed at 25%, and the needle-passing interval at 5s. Under these conditions, 188g of quinoa alkaloid extract was obtained. The polysaccharide content, protein content, total saponin content, total polyphenol content, and total flavonoid content of the quinoa alkaloid extract were measured to be 29.3%, 42.8%, 4.23%, 4.0%, and 1.6%.

[0043] Example 6:

[0044] Quinoa raw materials were washed with distilled water, drained, and dried in an oven at 60℃. The dried quinoa was then pulverized and passed through a 50-mesh sieve to obtain quinoa flour. 714.3g of quinoa flour was weighed and added to 10L of deionized water in an extraction tank. The pH of the extraction solution was adjusted to 8.5 with a food-grade calcium oxide mixture. The extraction temperature was set at 30℃, and extraction was carried out for 2.0 hours under these conditions. After extraction, the mixture was centrifuged at 6000 rpm for 10 minutes. The supernatant was collected and diluted to 20L with deionized water, then spray-dried. The inlet temperature was set at 128℃, the outlet temperature at 60℃~65℃, the fan speed at 97%, the peristaltic pump speed at 25%, and the needle-passing interval at 5s. Under these conditions, 210g of quinoa alkaloid extract was obtained. The polysaccharide content, protein content, total saponin content, total polyphenol content, and total flavonoid content of the quinoa alkaloid extract were measured to be 31.28%, 43.7%, 4.6%, 5.5%, and 2.1%, respectively.

[0045] Example 7:

[0046] Quinoa raw materials were washed with distilled water, drained, and dried in an oven at 60℃. The dried quinoa was then pulverized and passed through a 50-mesh sieve to obtain quinoa flour. 833g of quinoa flour was weighed and added to 10L of deionized water in an extraction tank. The pH of the extraction solution was adjusted to 8.5 with a food-grade calcium oxide mixture. The extraction temperature was set at 50℃, and extraction was carried out for 1.5 hours. After extraction, the mixture was centrifuged at 6000 rpm for 10 minutes. The supernatant was collected and diluted to 20L with deionized water before spray drying. The inlet temperature was set at 128℃, the outlet temperature at 60℃~65℃, the fan speed at 97%, the peristaltic pump speed at 25%, and the needle-passing interval at 5s. Under these conditions, 215g of quinoa alkaloid extract was obtained. The polysaccharide content, protein content, total saponin content, total polyphenol content, and total flavonoid content of the quinoa alkaloid extract were measured to be 32%, 44.78%, 5.0%, 5.7%, and 2.3%, respectively.

[0047] Example 8:

[0048] Quinoa raw materials were washed with distilled water, drained, and dried in an oven at 60℃. The dried quinoa was then pulverized and passed through a 50-mesh sieve to obtain quinoa flour. 714.3g of quinoa flour was weighed and added to 10L of deionized water in an extraction tank. The pH of the extraction solution was adjusted to 9.0 with a food-grade calcium oxide mixture. The extraction temperature was set at 40℃, and extraction was carried out for 2.0 hours under these conditions. After extraction, the mixture was centrifuged at 6000 rpm for 10 minutes. The supernatant was collected, diluted to 20L, and then spray-dried. The inlet temperature was set at 128℃, the outlet temperature at 60℃~65℃, the fan speed at 97%, the peristaltic pump speed at 25%, and the needle-passing interval at 5s. Under these conditions, 205g of quinoa alkaloid extract was obtained. The polysaccharide content, protein content, total saponin content, total polyphenol content, and total flavonoid content of the quinoa alkaloid extract were measured to be 31%, 42.7%, 4.3%, 5.3%, and 1.8%, respectively.

[0049] Example 9:

[0050] The quinoa raw materials were washed with distilled water, drained, dried in an oven at 60 °C, crushed, and passed through a 50-mesh sieve to obtain quinoa powder. Weigh 1000 g of quinoa powder, add 10 L of deionized water, and put it into an extraction tank. Adjust the pH of the extract to 8.5 with a food-grade calcium oxide mixture, set the extraction temperature to 40 °C, and extract for 2.5 h under this condition. After extraction, set the centrifuge speed to 6000 r / min and centrifuge for 10 min. Collect the supernatant, dilute it to 20 L, and then spray dry it. Set the inlet temperature to 128 °C, the outlet temperature to 60 °C - 65 °C, the fan speed to 97%, the peristaltic pump speed to 25%, and the needle passing interval to 5 s. Under this condition, 194 g of quinoa alkali extract was obtained. The polysaccharide content in the quinoa alkali extract was measured to be 29.6%, the protein content was 43.0%, the total saponin content was 4.8%, the total polyphenol content was 4.3%, and the total flavonoid content was 2.0%.

[0051] Example 10:

[0052] Anti-fatigue pharmacodynamic experiment

[0053] (1) Experimental grouping and administration

[0054] 120 SPF-grade inbred CD-1 male mice, weighing 20 ± 2 g, with the experimental animal license number: [SCXK(Beijing) 2021-0006], were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. After 1 week of adaptive feeding, the mice were randomly divided into 5 groups (n = 12 / group): blank control group, salidroside control group at 150 mg / kg, low-dose quinoa group at 200 mg / kg, medium-dose quinoa group at 400 mg / kg, and high-dose quinoa group at 800 mg / kg;

[0055] The quinoa alkali extract prepared in Example 7 was fully dispersed and dissolved with distilled water respectively, and at the same time, distilled water was used as the blank control group. After 28 days of intragastric administration, 6 mice in each group were randomly selected for the load-bearing swimming experiment, and 6 were used for the swimming fatigue experiment. During the feeding and administration period, the mice had free access to water and food, and the body weight of the mice was recorded. The experimental results are shown in Table 1. The results showed that during the 28-day intragastric administration period, all the mice grew well, and there were no significant differences among the five groups. However, compared with the quinoa administration dose group, at the same time, there were no abnormalities in the diet structure, mental state, and feces of the experimental group, indicating that the quinoa alkali extract has no significant toxic effect on mice.

[0056] Table 1 Body weight changes of mice over time

[0057]

[0058] (2) Organ index detection

[0059] All mice in the five groups were euthanized by cervical dislocation, and their hearts, livers, viscera, spleens, lungs, and kidneys were dissected. After rinsing with physiological saline and absorbing surface moisture with filter paper, the organs were weighed. The organ index was calculated as follows: Organ Index (%) = Organ Mass / Body Weight × 100%. The organ coefficient is an important indicator reflecting the development of animal organs and can directly reflect the toxic effects of the test substance on the animal. The effects of quinoa alkaloid extract on the organ coefficient of mice are shown in Table 2. The results showed no significant difference between the treatment group and the control group (P>0.05). This indicates that quinoa alkaloid extract does not cause organic lesions in mice.

[0060] Table 2. Effects of Quinoa Alkaloid Extract on Organ Coefficient in Mice

[0061]

[0062] (3) Mouse weight-bearing swimming test

[0063] The evaluation was conducted according to the method reported by Kim et al. (Kim J, Beak S, Ahn S, Moon BS, Kim BS, Lee SJ, Oh SJ, Park HY, Kwon SH, Shin CH, Lim K, Lee KP. Effects of taurine and ginseng extracts on energy metabolism during exercise and their anti-fatigue properties in mice. Nutr Res Pract. 2022, 16(1):33-45). Specifically, 30 minutes after the last administration, mice were fitted with lead weights equal to 2% of their body weight on their tails and swam in a swimming tank (approximately 50cm × 50cm × 40cm). The water depth was at least 30cm, and the water temperature was 25℃ ± 2.0℃. The time the mice remained submerged for the first 10 seconds after starting to swim was recorded; this was the weighted swimming time. The results are shown in Table 3. As shown in the table, the swimming time under load in the treatment groups was significantly improved compared with the blank control group. Among them, the medium dose group of quinoa alkaloid extract had the longest swimming time under load, which was highly significant (p<0.01). This indicates that quinoa alkaloid extract can effectively prolong the swimming time of mice and exert an anti-fatigue effect.

[0064] Table 3. Effects of Quinoa Alkaloid Extract on Weight-Bearing Swimming Time in Mice

[0065]

[0066] Note: p<0.05, p<0.01, compared with the blank control group.

[0067] (4) Blood lactate content determination

[0068] The specific method was as follows: 30 minutes after the last administration, the mice swam in the aforementioned swimming tank without load for 90 minutes. 200 μL of blood was collected from the orbital venous plexus, allowed to stand for 30 minutes, and then centrifuged at 1500 r / min for 15 minutes. The serum was collected for later use. Relevant detection reagents were added according to the lactate kit instructions, and the absorbance was measured using an ELISA reader to calculate the relevant content. The experimental results are shown in Table 4. As can be seen from the table, the lactate content in the blood of mice was reduced in both the administered groups and the blank control group. The quinoa medium-dose group had the lowest blood lactate content, which was highly significant (p<0.01). This indicates that quinoa alkaloid extract can accelerate the clearance of lactate accumulation, reduce lactate production, and thus alleviate physical fatigue.

[0069] The results of the experiment on the effect of quinoa alkaloid extract on blood lactate levels in mice are shown in Table 4. The results showed that the treated groups reduced blood lactate levels in mice compared with the blank control group, with the medium-dose quinoa group showing the lowest blood lactate levels, which was highly significant (p<0.01). This indicates that quinoa alkaloid extract can accelerate the clearance of lactate accumulation, reduce lactate production, and thus alleviate physical fatigue.

[0070] Table 4. Effects of Quinoa Alkaloid Extract on Blood Lactate Levels in Mice

[0071]

[0072] Note: p<0.05, p<0.01, compared with the blank control group.

[0073] (5) Determination of urea nitrogen content

[0074] Thirty minutes after the last administration, the mice swam in the aforementioned swimming tank without load for 90 minutes. 200 μL of blood was collected from the orbital venous plexus, allowed to stand for 30 minutes, and then centrifuged at 1500 r / min for 15 minutes. The serum was collected for later use. Relevant detection reagents were added according to the urea nitrogen kit instructions, and the absorbance was measured using an ELISA reader to calculate the relevant content. The results of the experiment on the effect of quinoa alkaloid extract on urea nitrogen content in mice are shown in Table 5. As can be seen from the table, the urea nitrogen content in the administered groups was reduced compared to the blank control group. The low-dose quinoa group had the lowest urea nitrogen content, which was highly significant (p<0.01). This indicates that quinoa alkaloid extract can reduce the production of serum BUN in the body, thereby alleviating fatigue.

[0075] Table 5 Effects of Quinoa Extract on Urea Nitrogen Levels in Mice

[0076] Note: p<0.05, p<0.01, compared with the blank control group.

[0077] (6) Determination of liver glycogen content

[0078] Mice were sacrificed immediately 30 minutes after the last administration. The liver was removed, rinsed with physiological saline, and blotted dry with filter paper. 0.1 g of liver and muscle tissue were accurately weighed, and 0.75 mL of glycogen extract was added to each. The mixtures were then ground at low temperature using a tissue homogenizer, followed by boiling for 20 minutes with three shaking cycles to ensure complete reaction. After cooling, the mixture was diluted to 5 mL with distilled water, mixed thoroughly, and centrifuged at 8500 rpm at room temperature. The supernatant was used as the glycogen extract. Glycogen content was determined using the anthrone-sulfuric acid method. The results of the effect of quinoa alkaloid extract on mouse liver are shown in Table 6. The results showed that the administered groups all increased liver glycogen content compared to the control group. Low, medium, and high doses of quinoa significantly increased liver glycogen content (p<0.01), indicating that quinoa alkaloid extract can improve liver glycogen storage.

[0079] Table 6. Effects of Quinoa Alkaloid Extract on Mouse Liver Function

[0080] Note: p<0.05, p<0.01, compared with the blank control group.

[0081] (7) HE staining of mouse muscle tissue

[0082] Tissues of skeletal muscle organs from mice in each experimental group were collected, fixed in 4% paraformaldehyde buffer, routinely embedded in paraffin, sectioned, stained with hematoxylin and eosin, and observed under a 100-degree optical microscope for pathological morphology. Figures 4-6 As shown, before exercise, the AE muscle fibers in each group were tightly arranged, intact in shape, and had uniform cell nuclei, with no obvious pathological changes. Figure 4 As can be seen, after exercise, the A2 muscle fibers in the blank control group were broken, the gaps were widened, the cell nuclei were swollen, and the local staining was uneven, indicating that exercise fatigue caused significant muscle damage. After exercise, the B2 muscle fibers in the rhodioloside control group had increased gaps and swollen cell nuclei, but the degree of damage was less than that in the blank control group. After exercise, the C2 muscle fibers in the low-dose quinoa group had local cracks and varying staining depths. Figure 5 After exercise, the degree of D2 muscle fiber tearing and cell nuclear swelling were reduced in the medium-dose quinoa group. Figure 6 After exercise, the E2 muscle fibers in the high-dose quinoa group were more neatly arranged, with no obvious nuclear swelling, and the degree of damage was significantly lower than that in the blank control group. It can be seen that with increasing dosage, the degree of muscle fiber tearing decreases, interstitial formation of muscle fibers lessens, and nuclear swelling decreases. The results show that quinoa alkaloid extract can alleviate skeletal muscle damage in fatigued mice.

[0083] Example 11:

[0084] Weigh out 12.0g of quinoa alkaloid extract powder, 25.0g of maltitol, 20.0g of coconut oil, 5.0g of skim milk powder, 8.0g of resistant dextrin, 0.8g of compound leavening agent, 0.5g of sea salt, 1.2g of lecithin, and 0.3g of natural vanillin. Sift all ingredients except coconut oil and lecithin and mix them evenly. Heat the coconut oil to 50℃ and add lecithin to emulsify. Mix the emulsion with the dry ingredients to form a dough, let it rise, roll it out, and bake it at 125℃ for 12 minutes. Crush the baked biscuits, add 15.0g of fructooligosaccharide syrup and 5.0g of coconut oil, mix and press into shape to obtain quinoa alkaloid extract compressed biscuits with anti-fatigue function.

[0085] Example 12:

[0086] Weigh out 20.0g of quinoa alkaloid extract, 65.0g of sorbitol, 10.0g of microcrystalline cellulose, 1.0g of magnesium stearate, and 0.5g of peppermint flavoring according to the following proportions: Quinoa alkaloid extract, sorbitol, and microcrystalline cellulose are passed through an 80-mesh sieve and mixed evenly; 5% ethanol solution is added for wet granulation, and the mixture is dried at 50℃ until the moisture content is ≤3%; magnesium stearate and peppermint flavoring are added and mixed evenly, and then compressed into 0.5g / tablet using a rotary tablet press at a pressure of 8kN; finally, hydroxypropyl methylcellulose is used for coating to increase the weight by 2%, thus obtaining anti-fatigue compressed candy.

[0087] Although embodiments of the invention have been disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, variations, and modifications are possible without departing from the spirit and scope of the invention and the appended claims. Therefore, the scope of the invention is not limited to the contents disclosed in the embodiments.

Claims

1. A method for preparing quinoa alkaloid extract with anti-fatigue activity, characterized in that... Produce according to the following steps: (1) Wash the quinoa raw material with deionized water, drain the water and dry it in an oven at 60°C. Then, pulverize and pass it through a 50-mesh sieve to obtain quinoa powder. (2) Add quinoa powder to the extraction tank, mix quinoa powder and deionized water at a ratio of 1:10 to 1:14, add 0.2% food-grade calcium oxide mixture to adjust the pH of the extract to 8.0 to 9.0, set the extraction temperature to 30℃ to 50℃, and extract for 1.5 to 2.5 hours under these conditions to obtain the extract; (3) Transfer the extract obtained in step (2) to a centrifuge for solid-liquid separation. Set the centrifuge speed to 6000 r / min and centrifuge for 10 min to obtain precipitate and supernatant. (4) After diluting the supernatant obtained in step (3) to twice the original volume, spray drying is carried out. The inlet temperature is controlled at 128℃, the outlet temperature is controlled at 60℃~65℃, the fan speed is controlled at 97%, the peristaltic pump speed is controlled at 25%, and the needle insertion interval is controlled at 5s to obtain quinoa alkaloid extract.

2. A quinoa alkaloid extract with anti-fatigue activity prepared by the method of claim 1, characterized in that: The quinoa alkaloid extract contains 40.2%–45.27% protein, 28.3%–32.37% polysaccharides, 3.5%–5.38% total saponins, 3.6%–6.0% total polyphenols, and 1.5%–2.4% total flavonoids.

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