Preparation method of pecan phospholipid capable of enhancing learning and memory ability

Abstract

The invention discloses pecan phospholipid of high-purity total phospholipid capable of enhancing learning and memory ability and a preparation method of the pecan phospholipid. The method comprises the following steps: 1) preparing a coarse phospholipid extract: mixing pecan cake meal with a solvent, performing ultrasonic treatment, recovering the solvent from an extracting solution under reduced pressure, standing for layering, removing an upper oil layer to obtain a lower extracting solution, extracting and washing with acetone, and drying to obtain the pecan coarse phospholipid extract; and 2) purification: dissolving the pecan crude phospholipid extract with an elution solvent, stirring a sample with silica gel, loading on a chromatographic column, eluting with the elution solvent, and collecting the eluent to obtain a chromatographic feed liquid. And (3) decolorizing: mixing the chromatography feed liquid with activated carbon, stirring, filtering, carrying out vacuum concentration on the filtrate, and carrying out vacuum low-temperature drying to obtain the pecan phospholipid with high-purity total phospholipid. The incubation period of mice entering a darkroom can be obviously prolonged by high and medium doses of the obtained pecan phospholipid, and the shuttle frequency of a high-dose group is obviously reduced; high dose of pecan phospholipid can obviously increase the incubation period of mouse step-down, and the step-down times of high and medium dose groups are also obviously reduced; the high and medium doses of pecan phospholipid can significantly shorten the platform discovery latency period of mice, and also can significantly increase the platform crossing frequency of mice. Therefore, the pecan phospholipid can enhance the learning and memory ability and can be used for preparing related products.

Description

Technical Field

[0001] The invention relates to the field of pecan processing, and in particular to a high-purity pecan total phospholipid extract (pecan phospholipid for short) capable of enhancing learning and memory abilities and a preparation method thereof. Background Art

[0002] Pecans are a species of the genus Carya in the family Julandaceae. Carya ) Perennial tall tree Carya serrata Carya illinoinensis (Wangenh.) K. Koch], native to North America, is one of the world's famous woody oil nuts. The pecan kernel is rich in healthy oils and has a high oil content, with an average oil content of about 60%, and some new varieties can be as high as 76%. The proportion of unsaturated fatty acids in the oil is high (more than 90%). Animal experiments and clinical experiments have shown that pecans have a good effect on reducing low-density lipoprotein and triglycerides in the blood, so they can play a role in lowering blood lipids, protecting blood vessels and heart health, and are recommended by the American Heart Association as "heart-protecting food". In addition to oil, pecans also contain about 10-15% high-quality plant protein, which has a complete range of essential amino acids and a reasonable ratio. It is a good raw material for preparing plant protein beverages. At present, oil extraction and preparation of protein beverages are the two main directions of deep processing of pecans. There are some patents and literature reports on pecan oil extraction and preparation of protein beverages.

[0003] The byproducts of pecan oil extraction and protein beverage preparation contain a valuable ingredient - phospholipids. Lecithin is a lipid containing phosphoric acid and belongs to complex polar lipids. Lecithin is the main component of biological membranes. Human nerve cells and brain cells are covered by cell membranes composed of phospholipids. Insufficient phospholipids can cause membrane damage, resulting in intellectual impairment and mental tension. The acetyl group contained in phospholipids enters the intercellular space and combines with choline to form acetylcholine. Acetylcholine is a signal molecule that transmits information between various nerve cells and brain cells. It can speed up the speed of information transmission between nerve cells and brain cells, enhance memory, and prevent Alzheimer's disease. Therefore, daily intake of a certain amount of phospholipids can enhance brain function, enhance brain memory and intelligence. At present, soybean lecithin, egg choline, walnut lecithin, etc. are widely used in industry. Compared with animal-derived lecithin, plant-derived lecithin has no risk of virus contamination, does not contain cholesterol, and is safer and healthier. The health effects of lecithin from different sources are different, and the differences mainly come from the different phospholipid components and fatty acid substitution chains contained. The inventor team's preliminary research found that the content of phosphatidylcholine in pecan lecithin is the highest (accounting for about 41% of the total phospholipids), followed by phosphatidylethanolamine (28%), phosphatidic acid (15%), phosphatidylserine (8%), phosphatidylglycerol (5%), and the lowest content is lysophosphatidylinositol (<0.1%). The proportion of phosphatidylcholine in pecan lecithin is relatively high, and it can be predicted that it has good development and utilization value.

[0004] The residue of pecans after oil removal or partial removal by squeezing, solvent extraction and other methods is pecan cake. Pecan cake is a good raw material for preparing protein beverages. The applicant team has previously determined that its phospholipid content is about 2-3%. In the preparation process of pecan protein beverages, it is necessary to remove the pigments and tannins in the seed coat to achieve the effects of decolorization and deastringency, and improve the color and taste of the final pecan protein beverage. The by-products of the decolorization / deastringency step are rich in phospholipids. Utilizing the by-products of the decolorization / deastringency process in the preparation process of pecan protein beverages to prepare high-content pecan total phospholipids can significantly increase the added value of the industry, rationally utilize precious resources, and effectively reduce environmental pollution. In the future, with the continuous development of the pecan deep processing industry, more phospholipid-rich by-products will inevitably be produced. The present invention provides an advanced preparation and application method for the rational development and utilization of these by-products, which is of great significance to the pecan industry.

[0005] Currently, no reports on the preparation method and application of phospholipids obtained from pecan processing by-products have been found. Summary of the invention

[0006] In order to overcome the shortcomings of the existing pecan processing method and improve the development and utilization rate of pecan, one of the purposes of the present invention is to provide a method for preparing pecan lecithin that can enhance learning and memory ability.

[0007] The second object of the present invention is to provide pecan lecithin with high purity total lecithin prepared by the above preparation method.

[0008] The third object of the present invention is to provide the use of the pecan lecithin in products that enhance learning and memory abilities, wherein the products are medicines, foods or health products.

[0009] The purpose of the present invention is achieved through the following technical solutions:

[0010] 1) Preparing crude lecithin extract: mixing pecan cake with an extraction solvent, performing ultrasonic extraction, filtering to obtain an extract, decompressing the extract to recover the solvent and then standing, discarding the upper oil layer after stratification to obtain a lower extract, extracting and washing with acetone, and drying the extracted and washed solution to obtain the pecan crude lecithin extract;

[0011] 2) Purification: dissolve the pecan lecithin extract with an elution solvent, mix the sample with silica gel, apply it to a chromatography column, elute with an elution solvent, collect the eluate, and obtain a chromatography feed solution;

[0012] 3) Decolorization: Mix the chromatography liquid with activated carbon, stir and filter, concentrate the filtrate under reduced pressure, and then dry under vacuum at low temperature to obtain pecan lecithin;

[0013] In the step 1), the extraction solvent is one or more of ethanol, methanol and dichloromethane, the solid-liquid ratio of cake to extraction solvent is 1:6~1:10 g / ml, the ultrasonic extraction conditions are: temperature 30±5℃, power 100W, time 30min, the number of acetone washings is 2 times, and the volume ratio of the extract to acetone is 1:0.3~1.

[0014] In the step 1), the drying method is to first concentrate to a small volume under reduced pressure at 30-60°C, and then dry at 0-30°C under vacuum.

[0015] In the step 2), the elution solvent is a mixed solvent of dichloromethane and methanol, with a volume ratio of 10:1 to 1:1.

[0016] In the step 2), the chromatography column is a silica gel chromatography column.

[0017] In the step 3), the ratio of the chromatography liquid to the activated carbon is 1 mL: 0.01-0.03 g, the stirring temperature is 15-30° C., the reduced pressure concentration temperature is 30-60° C., and the vacuum low-temperature drying temperature is 0-30° C.

[0018] Preferably, the method for preparing the pecan lecithin capable of enhancing learning and memory ability according to the present invention comprises the following steps:

[0019] 1) Preparation of crude lecithin extract: taking pecan cake, adding an extraction solvent mixed with one or more solvents selected from ethanol, methanol and dichloromethane, ultrasonic extraction, the solid-liquid ratio of cake to extraction solvent is 1:6-1:10 g / ml, ultrasonic extraction conditions: temperature 30 ± 5°C, power 100W, time 30min, filtering to obtain an extract, decompressing the extract to recover the solvent and then standing, discarding the upper oil layer after stratification to obtain a lower extract, extracting and washing with acetone twice, the volume ratio of the lower extract to acetone is 1:0.3-1, the liquid after extraction and washing is first concentrated to a small volume under reduced pressure at 30-60°C, and then vacuum-dried at 0-30°C, and the dried liquid is the pecan crude lecithin extract;

[0020] 2) Purification: using a mixed solvent of dichloromethane and methanol in a ratio of 10:1 to 1:1 as an elution solvent, dissolving the crude lecithin extract of pecans, mixing the sample with silica gel, applying it to a silica gel chromatography column, eluting with an elution solvent, collecting the eluate, and obtaining a chromatography feed solution;

[0021] 3) Decolorization: Mix the chromatography liquid with activated carbon in a ratio of 1 mL: 0.01-0.03 g. Stir at a temperature of 15-30°C. Filter after stirring. Concentrate the filtrate under reduced pressure at 30-60°C to a viscous state, and then dry at 0-30°C under vacuum to obtain pecan lecithin.

[0022] The present invention has the following advantages and beneficial effects:

[0023] (1) The preparation method provided by the present invention effectively utilizes the extract produced during the decolorization / deastringency process of the pecan protein beverage, which is a byproduct of the pecan processing industry. It achieves the purpose of high-value utilization, reduces environmental pollution, and has a wide range of applications.

[0024] (2) The method for preparing pecan lecithin provided by the present invention is to obtain the optimal extraction steps, solvents used and related parameters through a large number of experimental studies. The pecan lecithin is obtained by only three steps of preparing a crude lecithin extract, purifying and decolorizing. The operation is simple and easy. The extract contains high-purity total lecithin in pecans, and the total lecithin content reaches more than 50%.

[0025] (3) The pecan lecithin provided by the present invention contains effective ingredients such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine and phosphatidic acid, and does not contain cholesterol substances that may be contained in lecithin from animal sources. The fatty acids in the lecithin are mainly unsaturated fatty acids such as oleic acid and linoleic acid. It can significantly enhance learning and memory abilities, and can be used for people who wish to supplement lecithin during the growth and development period, those who use their brains a lot, or those who suffer from memory loss caused by physiological aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1The present invention provides a process flow chart for preparing pecan lecithin capable of enhancing learning and memory abilities. DETAILED DESCRIPTION

[0027] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0028] Example 1

[0029] A method for preparing pecan lecithin capable of enhancing learning and memory ability comprises the following steps:

[0030] 1) Preparation of crude pecan lecithin: 1 kg of pecan cake was mixed with 6 L of 85% ethanol and ultrasonicated at 25°C and 100 W for 30 min. The extract was decompressed to recover the solvent, and the upper oil layer was discarded to obtain the lower extract. The extract was extracted and washed twice with 30% acetone solvent. The extracted and washed liquid was concentrated to a small volume under reduced pressure and then vacuum dried at room temperature to obtain crude pecan lecithin.

[0031] 2) Purification: Dissolve the crude pecan lecithin in a mixed solvent of dichloromethane:methanol (10:1, v / v), mix the sample with silica gel, apply it to a silica gel chromatography column, and elute with a mixed solvent of dichloromethane-methanol in ratios of 10:1, 5:1 and 1:1 in sequence, collect the eluates, and combine them to obtain the chromatography feed solution.

[0032] 3) Decolorization: Add 0.03g / mL of activated carbon to the chromatography liquid, stir at 25℃, filter, and concentrate the filtrate at 60℃ to a viscous state under reduced pressure, and then vacuum dry at 30℃ to obtain pecan lecithin with a total lecithin content of 56.1%. The total lecithin content is determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0033] Example 2

[0034] A method for preparing high-purity pecan lecithin capable of enhancing learning and memory ability comprises the following steps:

[0035] 1) Preparation of crude pecan lecithin: 1 kg of pecan cake was mixed with 8 L of methanol and ultrasonicated at 30°C and 100 W for 30 min, and the extract was filtered to obtain the extract. The extract was decompressed to recover the solvent and then allowed to stand. After stratification, the upper oil layer was discarded to obtain the lower extract. The lower extract was extracted and washed twice with 50% acetone solvent. The extracted and washed liquid was decompressed and concentrated to a small volume, and then vacuum dried at room temperature to obtain crude pecan lecithin.

[0036] 2) Purification: Dissolve the crude pecan lecithin in a mixed solvent of dichloromethane:methanol (10:1, v / v), mix the sample with silica gel, apply it to a silica gel chromatography column, and elute with a mixed solvent of dichloromethane-methanol in a ratio of 10:1 and 3:1 in sequence, collect the eluates, and combine them to obtain the chromatography feed solution.

[0037] 3) Decolorization: Add 0.02g / mL of activated carbon to the chromatography liquid, stir at 15℃, filter, and concentrate the filtrate under reduced pressure at 30℃ to a viscous state, and then vacuum dry at 0℃ to obtain pecan lecithin with a total lecithin content of 54.8%. The total lecithin content is determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0038] Example 3

[0039] A method for preparing high-purity pecan lecithin capable of enhancing learning and memory ability comprises the following steps:

[0040] 1) Preparation of crude pecan lecithin: 1 kg of pecan cake was added with 10 L of a mixed solvent of dichloromethane and methanol (1:1, v / v), and then ultrasonicated at 35°C and 100 W for 30 min. The extract was filtered to obtain an extract. The extract was decompressed to recover the solvent and then allowed to stand. After stratification, the upper oil layer was discarded to obtain a lower extract. The lower extract was extracted and washed twice with 100% volume acetone solvent. The extracted and washed liquid was decompressed and concentrated to a small volume, and then vacuum dried at room temperature to obtain crude pecan lecithin.

[0041] 2) Purification of pecan lecithin: dissolve the crude pecan lecithin in a mixed solvent of dichloromethane:methanol (10:1, v / v), mix the sample with silica gel, apply it to a silica gel chromatography column, and elute it with a 4:1 mixed solvent of dichloromethane-methanol in sequence, collect the eluates, and combine them to obtain the chromatography feed solution.

[0042] 3) Decolorization: Add 0.01g / mL activated carbon to the chromatography liquid, stir at 30℃, filter, and concentrate the filtrate at 45℃ to a viscous state under reduced pressure, and then vacuum dry at 15℃ to obtain pecan lecithin with a total lecithin content of 53.7%. The total lecithin content is determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0043] Comparative Example 1

[0044] Pecan lecithin was extracted by the method described in the reported "CN 103665029 A method for preparing powdered soybean lecithin". 6 volumes of acetone were added to the pecan oil residue, stirred at room temperature for 30 minutes, and then allowed to stand for stratification. The acetone layer was discarded, and the water layer was retained and the above steps were repeated for extraction once, and the water layers were combined. After the water layer was decompressed to recover the solvent to a small volume, it was dried at low temperature in vacuum. The total lecithin content was determined to be 3.6%. The total lecithin content was determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0045] Comparative Example 2

[0046] A preparation method similar to that of Example 1 was used, except that the dichloromethane in the eluent was replaced with ethyl acetate. The steps are as follows:

[0047] 1) Preparation of crude pecan lecithin extract: 1 kg of pecan cake was mixed with 6 L of 85% ethanol and ultrasonicated at 25°C 100W for 30 min, and the extract was filtered to obtain an extract. The extract was decompressed to recover the solvent and then allowed to stand. After stratification, the upper oil layer was discarded, and the lower extract was separated and extracted and washed twice with 30% volume acetone solvent. The liquid after extraction and washing was decompressed and concentrated to a small volume, and then vacuum dried at room temperature to obtain crude pecan lecithin.

[0048] 2) Purification: Dissolve the crude pecan lecithin in a mixed solvent of ethyl acetate:methanol (10:1, v / v), mix the sample with silica gel, apply it to a silica gel chromatography column, and elute with a mixed solvent of ethyl acetate-methanol in ratios of 10:1, 4:1, and 1:1 in sequence, collect the eluates, and combine them to obtain the chromatography feed solution.

[0049] 3) Decolorization: Add 0.03g / mL of activated carbon to the chromatography liquid, stir at 25℃, filter, and concentrate the filtrate at 60℃ to a viscous state under reduced pressure, and then vacuum dry at 30℃ to obtain pecan lecithin with a total lecithin content of 39.0%. The total lecithin content is determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0050] Comparative Example 3

[0051] A similar preparation method as in Example 1 was used, using only ethyl acetate as the eluent. The steps are as follows:

[0052] 1) Preparation of crude pecan lecithin extract: 1 kg of pecan cake was mixed with 6 L of 85% ethanol and ultrasonicated at 25°C 100W for 30 min, and the extract was filtered to obtain an extract. The extract was decompressed to recover the solvent and then allowed to stand. After stratification, the upper oil layer was discarded to obtain a lower extract. The lower extract was extracted and washed twice with 30% acetone solvent. The extracted and washed liquid was decompressed and concentrated to a small volume, and then vacuum dried at room temperature to obtain crude pecan lecithin.

[0053] 2) Purification: dissolve the crude pecan lecithin with ethyl acetate, mix the sample with silica gel, apply it to a silica gel chromatography column, elute with ethyl acetate, collect the eluate, and combine them to obtain the chromatography feed solution.

[0054] 3) Decolorization: Add 0.03g / ml activated carbon to the chromatography liquid, stir at 25℃, filter, and concentrate the filtrate at 60℃ to a viscous state under reduced pressure, and then vacuum dry at 30℃ to obtain pecan lecithin with a total lecithin content of 30.7%. The total lecithin content is determined by "GB / T 5537-2008 Grain and Oil Inspection - Determination of Lecithin Content - Method 1".

[0055] Experimental Example 1

[0056] Evaluation of pecan lecithin in enhancing learning and memory

[0057] Experimental materials: Pecan lecithin prepared in Example 1 and commercially available soybean lecithin (total lecithin content greater than 95%) were ground with 0.2% CMC-Na to prepare a suspension for oral administration.

[0058] Experimental animals and feeding: ICR mice, SPF grade, 18-22g, male, Hangzhou Medical College Experimental Animal Center, production license number: SCXK (Zhejiang) 2019-0002. Mice were raised in the Animal Center of China Pharmaceutical University (Animal Use License Number: SYXK (Su) 2021-0011). The temperature of the breeding laboratory was 24±2℃; relative humidity was 60-80%; air exchange times per hour: 10-15 times / hour; light cycle: 12 (day) / 12 (night) hours, no more than 5 per cage. Feed: Complete mouse pellet feed, purchased from Yizheng Anlimao Biotechnology Co., Ltd., its quality meets GB14924.1-2010 "General Quality Standard for Experimental Animal Formula Feed". Drinking water: Drinking purified water.

[0059] Main reagents for the experiment: Scopolamine hydrobromide (S817762), 98%, Shanghai MacLean Biochemical Technology Co., Ltd.

[0060] The main experimental instruments: Morissa water maze (1056001), dark avoidance shuttle jumping platform experiment video analysis system (1056305), Beijing Zhongshidichuang Technology Co., Ltd.

[0061] Dosage setting: According to the literature and experimental material preparation process, the dosage of pecan lecithin samples is set at 120, 60, and 30 mg / kg; the dosage of soybean lecithin samples is set at 400 and 200 mg / kg.

[0062] Experimental animal grouping: 210 ICR mice were randomly divided into 7 groups after 1 week of adaptive feeding: (1) normal group; (2) model group; (3) soybean lecithin 400 mg / kg; (4) soybean lecithin 200 mg / kg; (5) pecan lecithin 120 mg / kg; (6) pecan lecithin 60 mg / kg; (7) pecan lecithin 30 mg / kg, with 30 mice in each group.

[0063] Experimental method: According to the above grouping, the test samples were gavaged once a day, with a dosing volume of 10mL / kg for 30 consecutive days. In the last few days of dosing, mice in each group started three behavioral trainings in advance according to different behavioral training standards. On the day of behavioral testing, 60 minutes after the last administration, 5 mg / kg of scopolamine was injected intraperitoneally to establish a memory impairment model. 30 minutes after the injection of scopolamine hydrobromide, 30 mice in each group were subjected to three behavioral tests: dark avoidance, platform jumping, and Morris water maze; 10 mice were used for each behavior.

[0064] (1) Dark-avoiding shuttle experiment method: The dark-avoiding apparatus consists of a bright room and a dark room. A 50W light bulb is hung above the bright room (20cm×20cm×20cm), and the bottom of the dark room (20cm×20cm×20cm) is covered with stainless steel rods with a diameter of 2mm, with a spacing of 1cm. Turn on the power switch and pass 36V AC. The two rooms are connected by a door that allows mice to enter and exit freely. First, place the mouse in the bright room to adapt for 10 seconds. When the mouse enters the dark room, turn on the power to give it an electric shock for 3 seconds, then turn off the power and the experiment is over. Mice that do not enter the dark room after 20 seconds are forced to enter and are shocked for 3 seconds. Repeat the test 24 hours later, and record the time (latency) when the mouse first enters the dark room from the bright room and the number of times it enters the dark room within 3 minutes.

[0065] (2) Jumping platform experiment method: The mouse jumping platform consists of a circular rubber platform (4.5 cm in diameter and 4.5 cm in height) and a floor composed of metal rods connected to an electric shock generator. When the mouse's limbs touch the metal floor, it will receive a 36V electric shock, while jumping onto the rubber platform will protect it from the electric shock. In the learning experiment, the mouse was placed on the rubber platform and allowed to adapt for 3 minutes. Then, it was placed on the metal floor, the power was turned on, and the electric shock was applied for 10 seconds. A test experiment was conducted 24 hours later, in which the mouse was placed on the rubber platform, and the time (latency) when it first jumped off the platform and the number of times it jumped off within 3 minutes were recorded.

[0066] (3) Morris water maze experimental method: The water depth in the maze was 25 cm (1 cm above the platform), the water temperature was (25±0.5)℃, and the circular platform was placed in the center of quadrant III. The mouse entered the water at any quadrant I, II, or IV, and the entry point was the midpoint of the quadrant. Each mouse was trained 3 times a day, each training session lasting 60 s, for 5 consecutive days. If the mouse found the platform within 60 s, it was allowed to stay on the platform for 10 s, and the time required for the mouse to find the platform from entering the water was recorded as the escape latency; if the mouse did not find the platform within 60 s, the operator guided it to the platform and it stayed there for 10 s, and its escape latency was recorded as 60 s.

[0067] Spatial exploration experiment: On the 6th day, the platform was removed and the mice entered the water from quadrant I for 60 s. The time when the mice first crossed the platform (latency) and the number of platform crossings were recorded.

[0068] Data processing and statistical methods: All data were expressed as mean ± standard error (M ± SE), and GraphPad Prism 6 software was used to perform ANOVA analysis to examine the significance of the inter-group comparisons.

[0069] Results of the mouse dark avoidance experiment: As shown in Table 1, compared with normal mice, the model group mice had a significantly shorter shuttle latency and a significantly increased shuttle frequency (P<0.01). Compared with the model group, high and medium doses of pecan lecithin significantly increased the latency of mice entering the dark room, and the shuttle frequency in the high-dose group was also significantly reduced (P<0.05). The latency and shuttle frequency in the high-dose soybean lecithin group were also significantly improved (P<0.05).

[0070] Table 1 Effects on latency and shuttle times in the dark avoidance test in mice (M±SE, N=10)

[0071]

[0072] **P<0.01, compared with the normal group; # P<0.05, compared with the model group.

[0073] Results of the mouse platform jumping experiment: As shown in Table 2, compared with normal mice, the model group mice had a significantly shorter platform jumping latency (P<0.01), and the number of shuttles was also significantly increased (P<0.05). Compared with the model group, high-dose pecan lecithin significantly increased the platform jumping latency of mice, and the number of steps off the platform in the high- and medium-dose groups was also significantly reduced (P<0.05). The latency and shuttle times of the high-dose soybean lecithin group were significantly improved, and the number of steps off the platform in the low-dose group was also significantly reduced (P<0.05).

[0074] Table 2 Effects on latency and number of steps off the platform in the mouse jumping test (M±SE, N=10)

[0075]

[0076] *P<0.05, **P<0.01, compared with the normal group; # P<0.05, compared with the model group.

[0077] Results of the Moriss water maze test in mice: As shown in Table 3, compared with normal mice, the model group mice found that the latency of the platform was significantly prolonged (P<0.01), and the number of crossing the platform was also significantly reduced (P<0.01). Compared with the model group, high and medium doses of pecan lecithin can significantly shorten the latency of mice to find the platform (P<0.01), and can also significantly increase the number of platform crossings of mice (P<0.05). The latency and platform crossing times of the high-dose soybean lecithin group were significantly improved, and the number of platform crossings of the low-dose group was also significantly increased (P<0.05).

[0078] Table 3 Effects on latency and platform crossing times in the Moriss water maze test in mice (M±SE, N=10)

[0079]

[0080] **P<0.01, compared with the normal group; # P<0.05, ## P<0.01, compared with the model group.

[0081] The above embodiments are only used to illustrate rather than limit the technical solutions of the present invention. Any changes, modifications, substitutions, combinations, and simplifications made under the spirit and principle of the present invention shall be equivalent replacement methods and shall be included in the protection scope of the present invention.

Claims

1. A method for preparing pecan lecithin capable of enhancing learning and memory ability, characterized in that: The following steps are involved: 1) Preparing crude lecithin extract: mixing pecan cake with an extraction solvent, performing ultrasonic extraction, filtering to obtain an extract, reducing the pressure on the extract to recover the solvent and then allowing the extract to stand, discarding the upper oil layer after stratification, separating the obtained lower extract, extracting and washing with acetone, and drying the extracted and washed solution to obtain the pecan crude lecithin extract; 2) Purification: dissolve the pecan lecithin extract with an elution solvent, mix the sample with silica gel, apply it to a chromatography column, elute with an elution solvent, collect the eluate, and obtain a chromatography feed solution; 3) Decolorization: Mix the chromatography liquid with activated carbon, stir and filter, concentrate the filtrate under reduced pressure, and then dry it at low temperature under vacuum to obtain pecan lecithin.

2. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: In the step 1), the extraction solvent is one or more of ethanol, methanol and dichloromethane, the solid-liquid ratio of cake to extraction solvent is 1:6~1:10 g / ml, the ultrasonic extraction conditions are: temperature 30±5℃, power 100W, time 30min, the number of acetone washings is 2 times, and the volume ratio of the extract to acetone is 1:0.3~1.

3. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: In the step 1), the drying method is to first concentrate to a small volume under reduced pressure at 30-60°C, and then dry at 0-30°C under vacuum.

4. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: In the step 2), the elution solvent is a mixed solvent of dichloromethane and methanol, with a volume ratio of 10:1 to 1:

1.

5. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: In the step 2), the chromatography column is a silica gel chromatography column.

6. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: In the step 3), the ratio of the chromatography liquid to the activated carbon is 1 mL: 0.01-0.03 g, the stirring temperature is 15-30° C., the reduced pressure concentration temperature is 30-60° C., and the vacuum low-temperature drying temperature is 0-30° C.

7. The method for preparing pecan lecithin capable of enhancing learning and memory ability as claimed in claim 1, characterized in that: The following steps are involved: 1) Preparation of crude lecithin extract: taking pecan cake, adding an extraction solvent mixed with one or more solvents selected from ethanol, methanol and dichloromethane, ultrasonic extraction, the solid-liquid ratio of cake to extraction solvent is 1:6-1:10 g / ml, ultrasonic extraction conditions: temperature 30 ± 5°C, power 100W, time 30min, filtering to obtain an extract, decompressing the extract to recover the solvent and then standing, discarding the upper oil layer after stratification to obtain a lower extract, extracting and washing with acetone twice, the volume ratio of the lower extract to acetone is 1:0.3-1, the liquid after extraction and washing is first concentrated to a small volume under reduced pressure at 30-60°C, and then vacuum-dried at 0-30°C, and the dried liquid is the pecan crude lecithin extract; 2) Purification: using a mixed solvent of dichloromethane and methanol in a ratio of 10:1 to 1:1 as an elution solvent, dissolving the crude lecithin extract of pecans, mixing the sample with silica gel, applying it to a silica gel chromatography column, eluting with an elution solvent, collecting the eluate, and obtaining a chromatography feed solution; 3) Decolorization: Mix the chromatography liquid with activated carbon in a ratio of 1 mL: 0.01-0.03 g. Stir at a temperature of 15-30°C. Filter after stirring. Concentrate the filtrate under reduced pressure at 30-60°C to a viscous state, and then dry at 0-30°C under vacuum to obtain pecan lecithin.

8. Pecan lecithin obtained by the preparation method according to any one of claims 1 to 7.

9. Use of the pecan lecithin as claimed in claim 8 in the preparation of a product for enhancing learning and memory abilities.

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