CN110433177B - DHA algae oil soft capsule and preparation method thereof - Google Patents
DHA algae oil soft capsule and preparation method thereof Download PDFInfo
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- CN110433177B CN110433177B CN201910788359.2A CN201910788359A CN110433177B CN 110433177 B CN110433177 B CN 110433177B CN 201910788359 A CN201910788359 A CN 201910788359A CN 110433177 B CN110433177 B CN 110433177B
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Abstract
The invention relates to the technical field of soft capsules, and in particular relates to a DHA algae oil soft capsule which comprises, by weight, 5-10 parts of DHA algae oil, 15-20 parts of fish oil, 3-6 parts of phosphatidylserine, 0.5-1.0 part of beeswax, 20-30 parts of gelatin, 20-30 parts of purified water, 10-15 parts of glycerol and 0.5-1.0 part of caramel color. The animal function test and the human body function test show that the invention has exact auxiliary memory improving function. The invention adopts modern preparation technology, is prepared by a reasonable quality control method and strict product management measures, and has stable quality and convenient taking.
Description
Technical Field
The invention relates to the technical field of soft capsules, and particularly relates to a DHA algae oil soft capsule and a preparation method thereof.
Background
Current products are primarily intended to supplement neurotransmitter precursors, potentiate their receptors or reduce their metabolism. However, neurotransmitters and receptors are widely distributed in various organs and tissues in the body, and side effects caused by these products are also systemic. Therefore, on the basis of understanding the nature of these diseases, the development of effective and low-side-effect health care products is urgently needed. And the health food industry in China develops rapidly, and new resource food becomes a hot spot of modern research.
DHA, docosahexaenoic acid, commonly known as NAOHUANGJIN, is a polyunsaturated fatty acid very important to human body, and belongs to an important member in Omega-3 unsaturated fatty acid family. And a large amount of data show that the health food containing docosahexaenoic acid (DHA) has the effects of enhancing immunity and assisting in improving memory, meanwhile, the DHA algae oil has important physiological functions in the aspects of the central nervous system, the cardiovascular system, the visual nervous system and the like, and the DHA is used as long-chain unsaturated fatty acid, so that the increase of the content of the DHA in a biological membrane has certain influence on the fluidity of the membrane, the trafficability of substances, and the activity of receptors, can improve the physiological function of the membrane, is favorable for enhancing the transmission of neural information, enhances the activity of the brain and the nervous system, and plays a very important role in maintaining the development and the function of the brain. DHA can also inhibit the occurrence, growth and metastasis of cancers, and regulate immune function by inhibiting the production of proinflammatory factors and regulating the expression of adhesion molecules, has been widely accepted by the society, and can be used as a nutrition enhancer and a food additive according to the regulation of DHA in various countries in the world. At present, DHA has been used in the fields of infant food, dairy products, beverages, soymilk, edible oil and fat and the like, and the market demand is very large, so that the production of DHA has become a development trend.
Meanwhile, many methods for improving the memory capacity of human or animals by adding a certain amount of DHA algal oil are disclosed in the prior art, and have certain effect, but the influence of the individual DHA algal oil on the memory capacity is limited, so that further improvement of the memory capacity on the basis of the DHA algal oil is a technical problem which needs to be solved at present.
Patent application CN107836717A discloses an algae oil DHA soft capsule with memory improving function and a preparation method thereof, wherein the algae oil DHA soft capsule comprises the following raw materials in parts by weight: 20-35 parts of DHA algae oil, 40-60 parts of gelatin, 50-75 parts of purified water, 10-18 parts of glycerol, 2-5 parts of light-shielding agent and 1.2-2.8 parts of preservative. The invention has scientific and reasonable design, can well control the hardness and the flexibility of the capsule shell, but can further improve the stability of the product and the performance of improving the memory.
Patent application CN 109259236a discloses a DHA algae oil soft capsule capable of enhancing memory and improving intelligence, which comprises a filling and a skin, wherein the filling comprises edible glucose, DHA algae oil, linseed oil, taurine and nutritional powder; the skin comprises edible glucose, edible gelatin, glycerol, sorbitol, citric acid, and water. The invention has the functions of promoting the development and growth of the brain and enhancing the immunity, and achieves the effects of enhancing the memory and improving the intelligence.
In addition, in the process of preparing the soft capsule by using the DHA algae oil, the problems of complicated preparation process, serious waste, low production efficiency, low yield and poor quality stability of finished products are easy to occur. In particular, the problems of low yield and poor stability caused by the improper experimental conditions in the preparation process of ingredients, sol and pressed pellets.
Patent application CN108477601A discloses a preparation method of DHA algae oil soft capsules, which is prepared by the processes of burdening, material dissolving, sol, pelleting, shaping, drying, packaging, detecting, warehousing and the like, and the whole process is processed in the same line, so that the nutrition loss caused by unstable factors is avoided, the influence of the environment on the capsules is reduced to the maximum extent, but the setting of the temperature and the degree of vacuum pumping in each step in the patent application can lead to the stability to be influenced to a certain degree, thereby influencing the yield.
Therefore, the technical problem to be solved at present is to provide a modern preparation process for preparing the soft capsule, and a reasonable quality control method.
Disclosure of Invention
The invention aims to overcome the defects of the existing soft capsule and application technology, and provides a DHA algae oil soft capsule and a preparation method thereof.
On one hand, the DHA algae oil soft capsule comprises, by weight, 5-10 parts of DHA algae oil, 15-20 parts of fish oil, 3-6 parts of phosphatidylserine, 20-30 parts of gelatin, 20-30 parts of purified water, 10-15 parts of glycerol and 0.5-1.0 part of caramel color.
Further, the phosphatidylserine is sieved by a 100-mesh sieve.
Further, 0.5-1.0 part of beeswax is also included.
Further, the health-care food comprises, by weight, 6-8 parts of DHA algal oil, 18-20 parts of fish oil, 4-6 parts of phosphatidylserine, 0.5-0.7 part of beeswax, 26-28 parts of gelatin, 25-28 parts of purified water, 12-14 parts of glycerol and 0.8-1.0 part of caramel color.
On the other hand, the invention provides a preparation method of the DHA algae oil soft capsule, which comprises the following steps;
(1) preparing materials: taking fish oil, phosphatidylserine, DHA algae oil and beeswax; heating 45-55% fish oil to 48-52 deg.C, adding Cera flava, and melting; cooling to 38-42 deg.C, adding DHA algae oil and the rest fish oil, mixing, adding phosphatidylserine, stirring, grinding, and filtering; vacuumizing; removing air bubbles to obtain material liquid;
(2) sol: heating purified water, glycerol and caramel color to 70-80 deg.C, adding glycerol and caramel color, stirring, adding gelatin, vacuumizing, removing bubbles, filtering, and keeping at 60 + -5 deg.C to obtain required gelatin solution;
(3) pelleting: pressing the prepared content material liquid and glue solution into pills, controlling the temperature to be 60 +/-5 ℃, the spraying temperature to be 37-42 ℃ and the humidity to be less than 45%;
(4) shaping: and (4) shaping the pressed soft capsule.
Further, the specific parameters of the sizing treatment in the step (4) are as follows: drying at 18-26 deg.C and humidity less than 45% for 4-5 hr.
Further, the method also comprises the following steps: washing pills; washing the shaped soft capsule with ethanol, and removing stains on the surface of the soft capsule.
Further, ethanol is 95% ethanol.
Further, the method also comprises the following steps: drying; transferring the cleaned soft capsule into drying chamber, controlling temperature at 18-26 deg.C and relative humidity less than 35%, and drying for 22-28 hr.
Further, the vacuum pumping range in the step (1) and the step (2) is-0.08 to-0.06 MPa.
Further, the filtration in the step (1) and the step (2) uses a 100-mesh sieve.
Furthermore, the invention also provides the steps of selecting pills, internally packaging, externally packaging, inspecting, warehousing and the like, and each link in the production process is strictly controlled, and the material entering a clean area is purified by 10 ten thousand grades in liquid preparation, sol, pill pressing, pill washing, drying, pill selection and packaging material storage rooms.
Furthermore, the pill selecting step comprises the steps of selecting out partial pills, leaking pills and removing unqualified products.
Further, the inner packaging step of the invention comprises the steps of filling the soft capsules into PET plastic bottles and sealing. Each bottle is filled with 60 granules.
Further, the outer packaging steps of the invention comprise labeling, boxing and boxing.
Furthermore, the inspection and warehousing of the invention is that each quality index specified by the product is inspected item by item respectively, and after the product is qualified, the product is sealed and warehoused.
The soft capsule provided by the invention is more suitable for filling oil medicines, and due to the sealing property of the capsule membrane, the soft capsule can block air, increase the stability of raw materials, well cover bad taste and is more convenient for patients to take.
Compared with the prior art, the invention has the beneficial effects that:
(1) the animal function test and the human body function test show that the invention has exact auxiliary memory improving function.
(2) The invention provides DHA algae oil, fish oil and phosphatidylserine as main components, and the DHA algae oil, the fish oil and the phosphatidylserine have synergistic effect, can improve the memory function, and has the advantages of effectiveness and low side effect.
(3) The soft capsule provided by the invention is prepared by adopting a modern preparation process under the conditions of a reasonable quality control method and strict product management measures, and has stable quality and convenient taking.
(4) In the preparation method provided by the invention, the two heating temperatures provided in the step of compounding make the stability of the capsule good; meanwhile, the degree of vacuum pumping provided by the invention further improves the stability of the capsule.
(5) The raw and auxiliary materials of the soft capsule provided by the invention are all non-infectious bacterium components, so that the quality of the product can be ensured.
(6) When the fish oil provided by the invention is mixed with the beeswax, the melting temperature is reduced, namely the temperature is not higher than 62-67 ℃ (the melting temperature of the beeswax), and the beeswax begins to dissolve.
Drawings
FIG. 1 is a schematic view of a water maze apparatus;
labeling: wherein, A, the first day training position B, the second day training position S and the starting point training
E1, E2, E3 and E4 are respectively a fault G and a ladder
Detailed Description
The technical solutions of the present invention will be further described with reference to specific examples, but the scope of the claims is not limited thereto.
Example 1 DHA algal oil soft capsule and preparation method thereof
A DHA algae oil soft capsule comprises, by weight, 8 parts of DHA algae oil, 20 parts of fish oil, 6 parts of phosphatidylserine, 28 parts of gelatin, 28 parts of purified water, 14 parts of glycerol, 1.0 part of caramel color and 0.6 part of beeswax. Wherein the phosphatidylserine is sieved by a 100-mesh sieve.
A method for preparing DHA algae oil soft capsules comprises the following steps: the method comprises the following steps: (1) preparing materials: weighing fish oil, phosphatidylserine, DHA algal oil and beeswax according to the formula ratio; heating 50% fish oil to 50 deg.C, adding Cera flava, and melting; cooling to 40 deg.C, adding DHA algae oil and the rest 50% fish oil, mixing, adding phosphatidylserine, stirring for 30 min, grinding the feed liquid with colloid mill for 3 times, and filtering with 100 mesh sieve; vacuumizing (-0.07Mpa) to remove bubbles to obtain material liquid; (2) sol: weighing gelatin, purified water, glycerol and caramel color according to the formula ratio, heating the purified water to 75 ℃, adding the glycerol and the caramel color, stirring for 30 minutes, adding the gelatin, continuously stirring for 1 hour, vacuumizing (-0.07Mpa) to remove bubbles, filtering by a 100-mesh sieve, and preserving heat at 60 ℃ to obtain the required glue solution for later use; (3) pelleting: placing the prepared content material liquid and glue solution on a soft capsule machine to be pressed into pills, controlling the temperature of a glue box to be 60 ℃, the temperature of a spraying body to be 40 ℃, the humidity to be 40 percent, and filling 0.6g of the content in each capsule; (4) shaping: shaping the pressed soft capsule (temperature 22 deg.C, humidity 38%, and drying for 4 hr). (5) Washing pills: washing the shaped soft capsule with 95% ethanol, and removing stains on the surface of the soft capsule. (6) And (3) drying: and transferring the cleaned soft capsule into a drying room, controlling the temperature at 22 ℃ and the relative humidity at less than 30%, and drying for 24 hours, wherein the dried soft capsule has no sticky and damp feeling. (7) Selecting pills: and (5) selecting partial pills and missed pills, and removing unqualified products. (8) Inner packaging: and (5) filling the soft capsules into a PET plastic bottle, and sealing. Each bottle is filled with 60 granules. (9) And (3) outer packaging: labeling, boxing and boxing. (10) And (6) inspection and warehousing: checking the quality indexes according to the specification of the product one by one, and stamping and warehousing after the quality indexes are qualified.
Example 2 DHA algal oil soft capsule and preparation method thereof
A DHA algae oil soft capsule comprises, by weight, 10 parts of DHA algae oil, 19 parts of fish oil, 5 parts of phosphatidylserine, 20 parts of gelatin, 20 parts of purified water, 10 parts of glycerol, 0.9 part of caramel color and 0.7 part of beeswax. Wherein the phosphatidylserine is sieved by a 100-mesh sieve.
A method for preparing DHA algae oil soft capsules comprises the following steps: the method comprises the following steps: (1) preparing materials: weighing fish oil, phosphatidylserine, DHA algal oil and beeswax according to the formula ratio; heating 45% fish oil to 52 deg.C, adding Cera flava, and melting; cooling to 38 deg.C, adding DHA algae oil and the rest 55% fish oil, mixing, adding phosphatidylserine, stirring for 30 min, grinding the feed liquid with colloid mill for 3 times, and filtering with 100 mesh sieve; vacuumizing (-0.06Mpa) to remove bubbles to obtain material liquid; (2) sol: weighing gelatin, purified water, glycerol and caramel color according to the formula ratio, heating the purified water to 70 ℃, adding the glycerol and the caramel color, stirring for 30 minutes, adding the gelatin, continuously stirring for 1 hour, vacuumizing (-0.08Mpa) to remove bubbles, filtering by a 100-mesh sieve, and preserving heat at 65 ℃ to obtain the required glue solution for later use; (3) pelleting: placing the prepared content material liquid and glue solution on a soft capsule machine to be pressed into pills, controlling the temperature of a capsule box to be 65 ℃, the temperature of a spraying body to be 42 ℃ and the humidity to be 38 percent, and filling 0.6g of the content in each capsule; (4) shaping: shaping the pressed soft capsule (temperature is 26 deg.C, humidity is 36%, and drying for 5 hr). (5) Washing pills: washing the shaped soft capsule with 95% ethanol, and removing stains on the surface of the soft capsule. (6) And (3) drying: the cleaned soft capsule is transferred to a drying room, the temperature is controlled at 18 ℃, the relative humidity is 25 percent, the drying is carried out for 28 hours, and the dried soft capsule has no sticky and damp feeling. (7) Selecting pills: and (5) selecting partial pills and missed pills, and removing unqualified products. (8) Inner packaging: and (5) filling the soft capsules into a PET plastic bottle, and sealing. Each bottle is filled with 60 granules. (9) And (3) outer packaging: labeling, boxing and boxing. (10) And (6) inspection and warehousing: checking the quality indexes according to the specification of the product one by one, and stamping and warehousing after the quality indexes are qualified.
Example 3 DHA algal oil soft capsule and preparation method thereof
A DHA algae oil soft capsule comprises, by weight, 5 parts of DHA algae oil, 15 parts of fish oil, 3 parts of phosphatidylserine, 0.5 part of beeswax, 30 parts of gelatin, 30 parts of purified water, 15 parts of glycerol and 0.5 part of caramel color. Wherein the phosphatidylserine is sieved by a 100-mesh sieve.
A method for preparing DHA algae oil soft capsules comprises the following steps: the method comprises the following steps: (1) preparing materials: weighing fish oil, phosphatidylserine, DHA algal oil and beeswax according to the formula ratio; heating 55% fish oil to 48 deg.C, adding Cera flava, and melting; cooling to 42 deg.C, adding DHA algae oil and the rest 45% fish oil, mixing, adding phosphatidylserine, stirring for 30 min, grinding the feed liquid with colloid mill for 3 times, and filtering with 100 mesh sieve; vacuumizing (-0.08Mpa) to remove bubbles to obtain content liquid; (2) sol: weighing gelatin, purified water, glycerol and caramel color according to the formula ratio, heating the purified water to 80 ℃, adding the glycerol and the caramel color, stirring for 30 minutes, adding the gelatin, continuously stirring for 1 hour, vacuumizing (-0.06Mpa) to remove bubbles, filtering by a 100-mesh sieve, and preserving heat at 55 ℃ to obtain the required glue solution for later use; (3) pelleting: placing the prepared content material liquid and glue solution on a soft capsule machine to be pressed into pills, controlling the temperature of a capsule box to be 55 ℃, the spraying temperature to be 37 ℃ and the humidity to be 38%, and filling the content in 0.6g per capsule; (4) shaping: shaping the pressed soft capsule (temperature is 18 deg.C, humidity is 36%, and drying for 5 hr). (5) Washing pills: washing the shaped soft capsule with 95% ethanol, and removing stains on the surface of the soft capsule. (6) And (3) drying: the cleaned soft capsule is transferred to a drying room, the temperature is controlled at 26 ℃, the relative humidity is 30 percent, the drying time is 22 hours, and the dried soft capsule has no sticky and damp feeling. (7) Selecting pills: and (5) selecting partial pills and missed pills, and removing unqualified products. (8) Inner packaging: and (5) filling the soft capsules into a PET plastic bottle, and sealing. Each bottle is filled with 60 granules. (9) And (3) outer packaging: labeling, boxing and boxing. (10) And (6) inspection and warehousing: checking the quality indexes according to the specification of the product one by one, and stamping and warehousing after the quality indexes are qualified.
Human body function test
Human body eating trial experiment for assisting in improving memory function
(1) Sample preparation: capsules nos. 1, 2, 3 and 4 are all provided by the Guangdong Tongde pharmaceutical Co., Ltd. The four are basically consistent in packaging, appearance, color and taste, wherein the capsule No. 1 is the DHA algae oil soft capsule provided by the embodiment 1 of the invention; the capsule No. 2 is the DHA algae oil soft capsule provided by the embodiment 2 of the invention; the capsule No. 3 is the DHA algae oil soft capsule provided by the embodiment 3 of the invention; capsule No. 4 is placebo. The recommended dose is 2 times daily, 2 capsules each time.
(2) Subject selection
Inclusion criteria were: the subjects were from a certain unit in Shanghai city, age 20-60 years, with no restriction. The health condition is good, no obvious brain, heart, liver, kidney and blood diseases exist, no long-term medicine taking history exists, similar tests (memory quotient and intelligence quotient tests) are not accepted, medicines or health-care foods related to memory improvement are not taken within one year, and the cooperation of voluntary subjects is guaranteed.
Exclusion criteria: pregnant or lactating women who are allergic to health foods; patients with serious diseases such as heart, liver, kidney and hematopoietic system; taking articles related to the tested function in a short time, and influencing the result judgment; if the test sample is not in compliance with the inclusion standard, the test sample is not eaten as specified, and the efficacy or the safety is not judged to be affected by the insufficiency of the test sample.
(3) Design and grouping of experiments
A control, double-blind, random design approach was used. Carrying out a first test before taking the sample, randomly dividing the sample into a test food group and a control group according to memory quotient, considering the balance of main factors influencing the result such as culture level, age and the like as much as possible to ensure comparability among the groups, and randomly extracting three groups as a test group 1, a test group 2 and a test group 3 after checking the balance of two groups of memory quotient, wherein the other group is the control group; each group had 52 subjects; the test group 1 took capsule No. 1, the test group 2 took capsule No. 2, the test group 3 took capsule No. 3, and the control group took capsule No. 4.
(4) Experimental methods
The test group and the control group start from 12 and 01 in 2018, and the samples are taken according to the recommended dose. The special person is responsible for taking samples and supervising administration. Health food or medicine related to memory improvement is not taken during the test period, and memory quotient or intelligence quotient test which is not related to the test is not participated. A second test was performed 45 days after the continuous administration.
(5) Index of efficacy
Testing using a clinical memory scale, testing the original score scale with each score after testing: pointing memory, associative learning, image free memory, meaningless graph recognization and portrait feature associative memory. And adding the test table points to obtain a total table point, and checking the memory quotient by using the total table point.
(6) Test equipment: a magnetic tape recording guide words and stimulating words pointing to memory and associative learning; picture material, picture freely remembering object picture, two groups are 15 pieces each, 30 pieces total (divide into A, two sets of B); re-recognizing pictures with meaningless figures, namely 20 pictures of target stimulation for first presentation, and 40 pictures of target stimulation for second re-recognition and 20 pictures of mixed stimulation pictures respectively, wherein the total number of the pictures is 60 (divided into two sets of pictures A and B); the portrait characteristics are related to the recall picture, 6 human faces are sketched in black and white for the first presentation, and 6 human faces are presented for the second recall, wherein the contents of the human faces and the human faces are the same, and the sequence is different, and the total number of the human faces is 12; the back of each portrait picture is marked with the characteristics of the portrait, such as surname, occupation, hobby and the like (divided into two sets, namely A and B). Recorder, stopwatch, recording paper.
(7) Test methods and requirements
The test method is that the front test and the back test of each test subject are performed by the same main tester, so that the system error is reduced.
And (3) testing time point: the test of each subject is carried out at the same time point, so as to avoid the influence of biological rhythm.
And in the second test, the main tester does not see the grouping list and tests in a blind method.
General requirements at test: testing the subject in a quiet room by trained personnel; except for the subject and the main tester, the presence of other people is avoided as much as possible; three sub-tests related to vision are arranged in the scale, and indoor light must ensure that a clear stimulating picture can be seen; the influence on memory performance caused by poor hearing or eyesight is eliminated as much as possible; attention must be paid to the mental state of the subject, and the test is carried out under the conditions that the subject is normal in mood, does not object to the test and is relatively concentrated in attention; whether the subject is tired, whether the attention is concentrated, whether the test is matched, whether the test is tense or not, whether the test is confident or not and the like are recorded on the first page of the recording paper; the test of the same subject requires to be finished at one time; when comparing point test performance with age scale points, care must be taken whether different point tests are performed under the same mental state. The original score of each score test result can be filled in the original score without error.
Data statistics
The test data is measured data, and two groups of test amount and memory quotient can be analyzed by t test. The self-contrast can adopt a paired t test, the parallel comparison between groups adopts a t test of the mean of two samples, the latter needs to carry out a homogeneity test of variance, carries out proper variable conversion on data with non-normal distribution or uneven variance, and carries out the t test by using the converted data after meeting the normality or the homogeneity of variance; if the converted data can not meet the requirement of normal variance, the t test or the rank sum test is adopted; but data with too large a coefficient of variation (e.g., CW > 50%) are tested using rank sum. And performing statistics by using INSTAT and SPSS statistical software.
Determination of results
On the premise that the two groups of memory quotient are balanced before the test, the memory quotient of the test food group after the test food is higher than that of the control group, the difference is significant, and the memory quotient of the test food group after the test food is higher than that of the memory quotient of the test food group before the test food is higher than that of the test food group before the test food is significant, so that the test sample can be judged to have the function of improving the memory function.
Influence of the invention on human memory efficacy index
TABLE 1 Effect of the invention on pointing to memory scores
| Before testing | After the test | |
| Control group | 22.27±3.49 | 23.41±4.76 |
| Test group 1 | 23.55±4.45 | 28.78±3.75*# |
| Test group 2 | 22.19±3.87 | 27.72±3.88*# |
| Test group 3 | 22.09±4.03 | 27.46±3.10*# |
< 0.05 compared to control; # compared to itself, # 0.05
The subject before the test is also the test subject;
as can be seen from Table 1, the directed memory content of the test group before sample taking is not significantly different from that of the control group (P is more than 0.05); after the sample is taken, the memory-oriented amount of the test group is obviously different from that of the control group and the test group before the sample is taken, wherein P is less than 0.05.
TABLE 2 influence of the invention on associative learning scale scores
| Before testing | After the test | |
| Control group | 26.26±5.35 | 27.18±4.59 |
| Test group 1 | 26.09±5.12 | 34.72±3.09*# |
| Test group 2 | 27.11±5.09 | 33.85±3.01*# |
| Test group 3 | 26.37±5.28 | 33.72±3.34*# |
< 0.05 compared to control; # compared to itself, # 0.05
As shown in Table 2, the associative study content of the test group before the sample taking was not significantly different from that of the control group ((P > 0.05), and the associative study content of the test group after the sample taking was significantly different from that of the control group and that of the test group before the sample taking itself ((P < 0.05)).
Table 3 influence of the invention on the image free recall score
| Before testing | After the test | |
| Control group | 23.34±4.85 | 23.42±4.66 |
| Test group 1 | 23.07±5.10 | 27.66±4.41*# |
| Test group 2 | 23.21±5.01 | 26.89±4.21*# |
| Test group 3 | 23.15±4.87 | 27.51±4.14*# |
< 0.05 compared to control; # compared to itself, # 0.05
As can be seen from Table 3, the image free recall ratio of the test group before sample taking was not significantly different from that of the control group (P >)
0.05); the image free memory content of the test group after sample taking is compared with that of the control group and the image free memory content before sample taking, and the difference is significant (P is less than 0.05).
Table 4 influence of the invention on meaningless graph re-recognition scores
| Before testing | After the test | |
| Control group | 15.47±5.43 | 15.44±5.69 |
| Test group 1 | 15.32±4.90 | 27.66±4.41 |
| Test group 2 | 15.44±4.98 | 26.80±4.58 |
| Test group 3 | 15.29±5.24 | 26.87±4.77 |
As can be seen from Table 4, the meaningless graph re-identification score of the test group before sample administration was not significantly different from that of the control group (P > 0.05); the meaningless graph re-identification amount of the test group after the sample was taken was not significantly different (P > 0.05) from that of the control group and that before the sample was taken.
TABLE 5 influence of the invention on the Portrait trait contact recall scale scores
| Before testing | After the test | |
| Control group | 18.96±5.47 | 19.40±4.52 |
| Test group 1 | 17.95±4.88 | 26.51±4.37*# |
| Test group 2 | 18.24±4.94 | 26.22±4.41*# |
| Test group 3 | 18.55±4.23 | 26.10±4.08*# |
< 0.05 compared to control; # compared to itself, # 0.05
As can be seen from Table 5, the human image characteristics of the test group before sample taking are compared with the recall value table and the control group, and no significant difference exists (P is more than 0.05); the human image characteristics of the test group after sample taking are compared with the memory scale before the control group and the self sample taking, and the significant difference exists (P is less than 0.05).
TABLE 6 Effect of the invention on memory quotient
| Before testing | After the test | |
| Control group | 96.74±11.48 | 98.45±11.32 |
| Test group 1 | 95.70±11.86 | 116.49±8.68*# |
| Test group 2 | 96.45±11.44 | 115.81±8.70*# |
| Test group 3 | 96.27±11.73 | 116.32±8.32*# |
< 0.05 compared to control; # compared to itself, # 0.05
As can be seen from Table 6, the memory quotient of the test group before sample taking was not significantly different from that of the control group (P > 0.05); the memory quotient of the test group after sample taking is obviously higher than that of the control group (P is less than 0.05); the memory quotient after the test group was taken was significantly higher than before the sample was taken (P < 0.05).
(8) Loss rate
After 45 days of experiments, 2 subjects in the control group and the test groups 1-3 are screened out because of taking the test articles discontinuously or being unable to judge the effect. Finally, the effective test population groups 1-3 and the control group are 52 cases, which are shown in Table 7.
TABLE 7 test loss
| Group of | Control group (example) | Test set 1 (example) | Test group 2 (example) | Test group 3 (example) |
| Before tasting | 54 | 54 | 54 | 54 |
| Number of loss | 2 | 2 | 2 | 2 |
| Loss rate | 3.70％ | 3.70％ | 3.70％ | 3.70％ |
The human body test results show that after the subject continuously takes the soft capsule provided by the invention for 45 days, the directional memory, the associative learning, the image free memory, the portrait characteristics, the memory scale and the memory quotient are improved and have significant difference (P is less than 0.05) compared with the control group and the subject before taking the sample. According to the evaluation standard of health food inspection and evaluation technical specification (2003 edition), the invention is prompted to have the function of assisting in improving the memory function of human body.
Functional test of animals
Animal experiment for assisting in improving memory function
(1) Materials and methods
Test article
The DHA algae oil soft capsule is from Guangdong Tongde pharmaceutical industry Co., Ltd, and the experiments in this part are carried out by adopting capsule contents, and are detected by Guangzhou disease prevention and control center.
(2) Laboratory animal
18-22g male SPF-grade Kunming mice were used as subjects, and were provided by the institutional laboratory animal center. Production license number of experimental animal: SCXK- (military) 2016-0050289652.
(3) Main Instrument Water maze (LW-II) institute of medicine, institute of Chinese medical science, mouse electro-optical stimulation Condition Refraction jumping platform-accurate North Zhenghua Bio-Instrument Equipment Co., Ltd
(4) Animal feeding
The experimental animals are bred in an SPF animal room, the temperature range of the animal room is 20-25 ℃, and the relative mixing degree is 40-70%. The feed is provided by Guangzhou university of traditional Chinese medicine, and the production license number is as follows: SCXK (Yue) 2016-
(5) Grouping of laboratory animals
Two large groups are designed in the experiment and are respectively used for a water maze experiment and a diving platform experiment. Each group of 20 mice is divided into a control group and a test group 4, wherein the test group 4 uses the DHA algae oil soft capsules (10 mice in each group) in the embodiment 1, the dosage is 0.30g/kg & BW respectively, namely 3.0g of DHA algae oil soft capsule samples are weighed, peanut oil is added to 200mL, the mice in the low, medium and high dosage groups are respectively subjected to oral gavage, the gavage amount is 0.2mL/10g & BW, the control group is given the same amount of peanut oil, and the control group is subjected to the continuous gavage once a day for 30 days.
And (4) performing a diving platform experiment and a water maze experiment respectively the next day after the last intragastric administration, and recording corresponding indexes.
(6) Diving platform experiment
The next day after the last gavage. The animals were placed in a reaction chamber (on-table, under-table) and acclimated for 3min, then placed on a copper grid in the reaction chamber and immediately supplied with 36v ac. The animal is subjected to an electric shock, the normal response of which is to jump back to the platform (insulator) to avoid injurious stimuli. Most animals may jump to the cupboards again or many times, and quickly jump back to the platform after receiving an electric shock. After training once, the animals are placed on the platform of the reaction box, and the error times of each mouse jumping off the platform and the latency of the first jumping off the platform within 5min are recorded as the learning achievement. And performing a retest after 24h or 48 h. Memory regression experiments after 5 days with training stopped.
The latency for the first time each mouse jumped off the platform, the number of shocks 3min for each mouse, and the total number of shocked animals were recorded, along with the percentage of animals that had a false response (the number of shocked animals was a percentage of the total number of animals in the group).
(7) Water maze experiment
The last gavage began training the day after the last time, once a day. The training time of the mice was defined as 120s, and all mice that did not reach the end point within 120s were scored as 120 s.
The water maze swimming box is made of polyethylene plastic. The swimming box is 100cm long, 100cm wide, 30cm high, 90cm long, 90cm wide and 30cm high in inner diameter, 12cm wide, lanes are fixed in the moving direction (as shown in the following figure), the labyrinth lanes are trained once every day with the water depth of 15cm and the water temperature kept at 25 +/-1 ℃, the training is continued for 5 days, the mouse is placed near a ladder (G) in the first training, the mouse is automatically climbed for 3 times, and the mouse is placed near the ladder before each training and is automatically climbed for one time; carrying out experiments in stages; the distance is gradually increased according to the learning performance of the mice. The first day of training begins with the stop at gear death at a. The next day training was started at B, and this course was 3 times, until more than 80% of the mice reached the end point (G). Training was started on day 5 from the start (S) and finally evaluated for 5 days of total learning performance (i.e. number of mice to end, number of errors per 5 balances and time to end). Memory regression experiments were performed after 1 week.
Referring to fig. 1, the total number of errors, total time to endpoint and total number of animals to endpoint (percentage) within 2min were calculated for each group of animals for 5 training and testing.
Both the diving platform experiment and the water maze experiment were performed twice.
(8) Statistical analysis method
Statistical analysis was performed using SPSS for Windows software. The difference of multiple groups of measurement indexes adopts variance analysis: and performing proper variable transformation or a rank sum test on the data with abnormal distribution or variance. The difference of the counting indexes is detected by a chi-square method: and when the total number of the cases is less than 40 or the total number of the cases is equal to or more than 40 but the theoretical number is equal to or less than 1, applying an exact probability method.
(9) Determination of results
Any one of the incubation period, the error frequency and the number of animals jumping off the platform within 3min in the platform jump experiment is positive, and the experiment can be judged to be positive. In the water maze experiment, any index of the time reaching the end point, the error frequency and the number of animals reaching the end point is positive, and the test can be judged to be positive. Any one of the water maze experiment and the diving platform experiment is positive in result, and the repeated experiment results are consistent (the results of two times of the same repeated experiment are positive), so that the DHA algae oil soft capsule can be judged to be positive in animal experiment result for assisting in improving memory function.
(10) Influence of DHA algae oil soft capsule on improving memory function of mice
Diving platform experiment
First time experiment result
As can be seen in Table 8, the test incubation period of mice in test group 4 was longer than that of the control group in the mouse jump bench test, and the difference was statistically significant (P < 0.05).
TABLE 8 influence of DHA algal oil soft capsules on the incubation period of the mouse diving platform experiment
Second experimental results
As can be seen from Table 9, in the mouse jump bench test, the test incubation period of mice in test group 4 was longer than that in the control group, and the difference was statistically significant (P < 0.05).
TABLE 9 influence of DHA algal oil soft capsules on the incubation period of the mouse diving platform experiment
The results of the two experiments are consistent, namely in the mouse jump experiment, the test latency period of the mice of the test group 4 is longer than that of the control group, and the difference has statistical significance (P is less than 0.05),
water maze experiment
First time experiment result
As can be seen from Table 10, in the water maze experiment, the animals of test group 4 reached the endpoint longer than the control group, and the difference was statistically significant (P < 0.05).
TABLE 10 DHA algae oil soft capsulesEffect on time to endpoint in mouse Water maze experiments
Second experimental results
As can be seen from Table 11, in the water maze experiment, the animals of test group 4 reached the endpoint longer than the control group, and the difference was statistically significant (P < 0.05).
TABLE 11 influence of DHA algal oil soft capsules on the time to endpoint in the mouse Water maze experiment
The results of both experiments were consistent, i.e. animals in test group 4 reached the endpoint longer than the control group and the difference was statistically significant (P < 0.05).
In the experiment, mice are fed with the DHA algal oil soft capsules (0g/kg & BW, 0.30g/kg & BW) at the following doses for 30 days, and then a diving platform experiment and a water maze experiment are respectively carried out. The results show that the diving platform experiment latency of the mice of the dosage group in the DHA algae oil soft capsule is prolonged, and the time for the water maze experiment of the mice of the dosage group in the DHA algae oil soft capsule to reach the end point is shortened. The DHA algae oil soft capsule can be preliminarily judged to have the function of assisting in improving memory.
Comparative example 1
A soft capsule with memory improving function comprises a capsule shell and contents filled in the capsule shell; the content is prepared from the following raw materials in parts by weight: 8 parts of DHA algal oil and 0.7 part of beeswax; the capsule shell is prepared from the following raw materials in parts by weight: 28 parts of gelatin, 28 parts of purified water, 14 parts of glycerol and 1.0 part of caramel color.
Comparative example 2
A soft capsule with memory improving function comprises a capsule shell and contents filled in the capsule shell; the content is prepared from the following raw materials in parts by weight: 20 parts of fish oil and 0.7 part of beeswax; the capsule shell is prepared from the following raw materials in parts by weight: 28 parts of gelatin, 28 parts of purified water, 14 parts of glycerol and 1.0 part of caramel color.
Comparative example 3
A soft capsule with memory improving function comprises a capsule shell and contents filled in the capsule shell; the content is prepared from the following raw materials in parts by weight: 6 parts of phosphatidylserine and 0.7 part of beeswax; the capsule shell is prepared from the following raw materials in parts by weight: 28 parts of gelatin, 28 parts of purified water, 14 parts of glycerol and 1.0 part of caramel color.
Comparative example 4
A soft capsule with memory improving function comprises a capsule shell and contents filled in the capsule shell; the content is prepared from the following raw materials in parts by weight: 8 parts of DHA algae oil, 5 parts of fish oil, 6 parts of phosphatidyl serine and 0.7 part of beeswax; the capsule shell is prepared from the following raw materials in parts by weight: 28 parts of gelatin, 28 parts of purified water, 14 parts of glycerol and 1.0 part of caramel color.
Detection example 1: method for testing influence on learning and memory by diving platform method
1. Experimental animals: healthy and qualified NIH mice are half as male and female.
2. An experimental instrument: DT-200 diving tower automatic tester (Shanghai Xinman scientific Equipment Co., Ltd.).
3. Experimental groups: the soft capsules of example 1 and comparative examples 1 to 4 were fed at the same time in normal feeding, and the experimental dose: the dosage is 5g/Kg twice a day. The blank control group was fed normally without any drug.
4. Experimental methods
4.1 experimental animal feeding and grouping: selecting healthy and qualified NIH mice, randomly grouping, feeding 20 mice in each group with half of male and female in cages, and continuously feeding for 30 days according to the experimental dose, wherein the animal weight is 18 +/-2 g.
4.2 diving platform experiment
4.2.1 diving platform experimental principle: in an open space, animals move mostly around the edges and corners. A high platform is arranged in the center of the square space, a copper grid is laid at the bottom of the high platform, and the copper shed is electrified. When the animal is placed on the platform it jumps off the platform almost immediately and explores around. If the animal is shocked when jumping off the platform, its normal response is to jump back to the platform to avoid injurious stimuli. Most animals may jump to the copper again or many times and quickly jump back to the platform after receiving an electric shock.
4.2.2 diving platform experiment training: the platform jumping device is a long hair body, a mouse is placed on a copper grid during an experiment, when the copper grid is electrified, the mouse jumping on the copper grid is subjected to electric shock, the normal reaction is to avoid the electric shock to jump up to a platform, most of mice are likely to jump down to the platform again or for multiple times and jump back to the platform rapidly after receiving the electric shock, and the training is carried out in such a way. The training frequency was 1 per 2 days, and the training time was 15 times during the gavage period, which was 1 hour before gavage. The specific training scheme is as follows: and controlling the voltage to be 36V, putting the mouse on a copper net, adapting to the environment for 3min, then electrifying, jumping up the platform to avoid electric shock after the mouse is shocked, and regarding that the mouse touches the copper net simultaneously with two feet as electric shock if the mouse jumps down the platform as an error response. After 30 days of gavage, the test was performed. When the test is started, the mouse is placed on a diving platform, a stopwatch is pressed at the same time, the 1 st time of the mouse jumping down (namely the electric shock volt period) is recorded, the mouse jumping down times (namely the error times) within 5min are recorded and used as a memory index, the average value of the mouse jumping down times is taken, and the specific data are shown in table 1.
TABLE 12 influence of mouse learning and memory test data
| Electric shock latency (second) | Number of errors within 5min | |
| Blank control group | 176 | 4.1 |
| Example 1 | 286 | 1.4 |
| Comparative example 1 | 254 | 1.9 |
| Comparative example 2 | 249 | 1.8 |
| Comparative example 3 | 252 | 2.1 |
| Comparative example 4 | 262 | 1.7 |
And (3) analyzing an experimental result: table 12 shows the data of the test on the effect of learning and memory in mice, and the electric shock latency and the number of errors in 5min are mainly recorded, and it can be seen from the experimental data that the electric shock latency of the mice can be prolonged in example 1 and comparative examples 1 to 4 compared with the blank control group. In addition, both example 1 and comparative examples 1-4 resulted in a significant reduction in the number of errors in mice within 5min compared to the blank control. In addition, the effect of example 1 is more significant than that of comparative examples 1 to 4.
Stability test
1.1 instruments
LHH-150GSP synthetic drug stability test chamber (Shanghai Lanbao test Equipment Co., Ltd.); an electric heating air-blast drying oven (Shanghai-Hengscientific instruments Co., Ltd.).
1.2 reagent
Soft capsules: specification: 0.6 g/pellet, the product obtained in example 1 of the production method of example 1, the product obtained in example 2 of the production method of example 2, the product obtained in example 3 of the production method of example 3, the product obtained under the production method of comparative example 5, and the product obtained under the production method of comparative example 6.
2 methods and results
Adopting methods such as long-term test investigation and the like to investigate the items: sensory requirement, physicochemical index, microorganism index, index of marker component, and loading difference.
2.1 Long term test
The test article (the product obtained by the production method of example 1, example 2, example 3, comparative example 5, and the product obtained by the production method of comparative example 6) was left at 25 ℃ 2 ℃ and 60% ± 10% relative humidity for 24 months, and sampled and inspected at 3 months, 6 months, 9 months, 12 months, 18 months, and 24 months, respectively. The test items and results are shown in tables 13 to 17.
The long-term test result shows that the soft capsule is placed for 24 months under the conditions of the temperature of 25 +/-2 ℃ and the relative humidity of 60 +/-10 percent and sampled for detection, the result is compared with 0 month, various indexes are not obviously changed, and the measured data is stable.
Comparative example 5
The difference from the preparation method of the soft capsule in example 1 is: the steps (1) and (3) are the same as those in example 1, and the formula is also the same as that in example 1.
In the step (1), preparing materials: weighing fish oil, phosphatidylserine, DHA algal oil and beeswax according to the formula ratio; heating 50% fish oil to 65 deg.C, adding Cera flava, and melting; cooling to 35 deg.C, adding DHA algae oil and the rest 50% fish oil, mixing, adding phosphatidylserine, stirring for 15 min, grinding the feed liquid with colloid mill for 2 times, and filtering with 100 mesh sieve; vacuumizing (-0.07Mpa) to remove bubbles to obtain material liquid;
(3) pelleting: placing the prepared content material liquid and glue solution on a soft capsule machine to be pressed into pills, controlling the temperature of a glue box to be 53 ℃, the temperature of a spraying body to be 30 ℃, the humidity to be 45 percent, and filling 0.6g of the content in each capsule;
comparative example 6
The difference from the preparation method of the soft capsule in example 1 is: the steps (1) and (2), and the rest of the steps are the same as those of the example 1, and the formula is also the same as that of the example 1.
In the step (1), preparing materials: weighing fish oil, phosphatidylserine, DHA algal oil and beeswax according to the formula ratio; heating 50% fish oil to 50 deg.C, adding Cera flava, and melting; cooling to 40 deg.C, adding DHA algae oil and the rest 50% fish oil, mixing, adding phosphatidylserine, stirring for 20 min, grinding the feed liquid with colloid mill for 2 times, and filtering with 100 mesh sieve; vacuumizing (-0.04Mpa) to remove bubbles to obtain material liquid;
in the step (2), sol: weighing gelatin, purified water, glycerol and caramel color according to the formula ratio, heating the purified water to 80 ℃, adding the glycerol and the caramel color, stirring for 20 minutes, adding the gelatin, continuously stirring for 1 hour, vacuumizing (-0.05Mpa) to remove bubbles, filtering by a 100-mesh sieve, and preserving heat at 65 ℃ to obtain the required glue solution for later use;
Claims (6)
1. a preparation method of a DHA algae oil soft capsule is characterized by comprising the following steps;
(1) preparing materials: taking 18-20 parts of fish oil, 4-6 parts of phosphatidylserine, 6-8 parts of DHA algae oil and 0.5-0.7 part of beeswax; heating 45-55% fish oil to 48-52 deg.C, adding 0.5-0.7 part Cera flava, and melting; cooling to 38-42 deg.C, adding 6-8 parts of DHA algae oil and the rest fish oil, mixing, adding 4-6 parts of phosphatidylserine, stirring, grinding, and filtering; vacuumizing; removing air bubbles to obtain material liquid;
(2) sol: taking 25-28 parts of purified water, 12-14 parts of glycerol and 0.8-1.0 part of caramel color, heating 25-28 parts of purified water to 70-80 ℃, adding 12-14 parts of glycerol and 0.8-1.0 part of caramel color, stirring, adding 26-28 parts of gelatin, vacuumizing, removing bubbles, filtering, and keeping the temperature at 60 +/-5 ℃ to obtain the required glue solution for later use;
(3) pelleting: pressing the prepared content material liquid and glue solution into pills, controlling the temperature to be 60 +/-5 ℃, the spraying temperature to be 37-42 ℃ and the humidity to be less than 45%;
(4) shaping: and (4) shaping the pressed soft capsule.
2. The method for preparing DHA algae oil soft capsules according to claim 1, wherein the specific parameters of the sizing treatment in the step (4) are as follows: drying at 18-26 deg.C and humidity less than 45% for 4-5 hr.
3. The method for preparing the DHA algae oil soft capsule according to claim 1, further comprising the following steps: washing pills; washing the shaped soft capsule with ethanol.
4. The method for preparing the DHA algae oil soft capsule according to claim 1, further comprising the following steps: drying; transferring the cleaned soft capsule into drying chamber, controlling temperature at 18-26 deg.C and relative humidity less than 35%, and drying for 22-28 hr.
5. The method for preparing DHA algae oil soft capsules according to claim 1, wherein the vacuum pumping range in the steps (1) and (2) is-0.08 to-0.06 MPa.
6. The method for preparing DHA algae oil soft capsules according to claim 1, wherein the filtering in step (1) and step (2) is performed with 100 mesh sieve.
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