CN114145347A - Phospholipid composition capable of improving learning and memory ability and application thereof - Google Patents

Phospholipid composition capable of improving learning and memory ability and application thereof Download PDF

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CN114145347A
CN114145347A CN202111453109.7A CN202111453109A CN114145347A CN 114145347 A CN114145347 A CN 114145347A CN 202111453109 A CN202111453109 A CN 202111453109A CN 114145347 A CN114145347 A CN 114145347A
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composition
phospholipid
amount
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food
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王雯丹
吉塞拉·阿德里安娜·怀斯
司徒文佑
叶文慧
刘彪
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Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
Inner Mongolia Dairy Technology Research Institute Co Ltd
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00Milk preparations; Milk powder or milk powder preparations
    • A23C9/152Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1528Fatty acids; Mono- or diglycerides; Petroleum jelly; Paraffine; Phospholipids; Derivatives thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12Fatty acids or derivatives thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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Abstract

The invention provides a phospholipid composition capable of improving learning and memory abilities and application thereof, wherein the phospholipid composition comprises Dihydrosphingosine (DSPH), Phosphatidylcholine (PC) and Phosphatidylinositol (PI), and the total phospholipid mass in the composition is 100 percent: the amount of sphinganine in the composition is 2% -10%; the amount of phosphatidylcholine in the composition is 20% to 59%; the amount of phosphatidylinositol in the composition is 3% to 15%. The phospholipid composition can effectively improve the learning and/or memory ability of organisms, and can be used for preparing various health foods and health foods such as infant formula powder, supplementary food or nutritional supplements and the like.

Description

Phospholipid composition capable of improving learning and memory ability and application thereof
Technical Field
The invention relates to a phospholipid composition and application thereof, in particular to a phospholipid composition containing Dihydrosphingosine (DSPH), Phosphatidylcholine (PC) and Phosphatidylinositol (PI) and application thereof in improving learning and/or memory capacity, belonging to the technical field of lipid.
Background
Phospholipids are a complex group of lipids. It is an important component of natural cell membrane structure, and its amphiprotic characteristics are derived from its hydrophobic tail and hydrophilic head. This property affects its role and function in the organism. They belong to the group of polar lipids, which are defined as lipids containing phosphorus, as they are literally defined. Polar lipids, which are the basic components of milk, can be used to emulsify fat in water because, along with proteins, they are the main component of the milk fat globule membrane, encapsulating the fat droplets secreted by mammary cells. Like other biofilm components, milk fat globule membranes contain, in addition to phospholipids, glycoproteins, glycolipids (such as cerebrosides and gangliosides), total and partial glycerols, free fatty acids and cholesterol, and the like.
In dairy products, glycerophospholipids and sphingolipids are the most important phospholipids in high amounts. The phospholipids account for about 0.5-1% of milk fat, and about 60-70% of the phospholipids in milk are located on the milk fat globule membrane, mainly on the outer bilayer membrane, including sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, etc. The origin of phospholipids, like the milk fat globule membrane, is the superficial plasma membrane of mammary secretory cells. Scientists hypothesize that most phospholipids, including those containing choline, and sphingomyelin, are located on the outside of the membrane. And phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol are located on the inside surface of the membrane.
Phospholipids play an important role in different cellular physiological processes. Phospholipid metabolism plays an important role in regulating intercellular signal transduction, proliferation and apoptosis, and systemic response to inflammation. There is evidence that phospholipids have major health benefits, including modulating immune responses, some prophylactic and therapeutic effects on some cancers, and inhibition of cholesterol absorption; has good regulation and control effects on cardiovascular and cerebrovascular diseases, inflammation and gastrointestinal infection, pressure, nervous system myelination, neurodevelopment and the like. The phospholipids have good emulsifiability and can be used as delivery systems of fat-soluble substances.
The total fat in the colostrum is about 2% of the total mass, and in the mature milk, the total fat is increased to 3-5% of the total mass. There were some differences between the studies, but the phospholipid concentration in colostrum was usually highest, with the total phospholipid content gradually decreasing with the lactation period. Studies have shown that diet, time of delivery, genetic factors and even the sex of the infant, etc., may affect the composition of the milk fat globule membrane in human milk. Human milk was analyzed by Benoit et al and Garcia et al sequentially and was found to contain lower phosphatidylethanolamine and higher sphingomyelin than other animal milks. In addition Garcia et al found that human milk is more abundant in phospholipids than other species, especially phosphatidylethanolamine plasmalogens.
In recent years, complex lipids contained in the milk fat globule membrane in human milk or bovine milk-based infant formula have been considered to play an important role in infant brain development and cognition. There is currently no unified understanding of how complex lipids in the diet participate in the process of cognitive development and its mechanism, but it is hypothesized that there may be three reasons: (1) the human brain is an organ composed of lipids, complex lipids such as phospholipids and gangliosides constitute 35% of the total lipid of the brain; (2) complex lipids such as gangliosides, etc., are rapidly ingested and utilized by brain development during the fourth month of pregnancy to the 4-5 years of childhood; (3) human milk fat globule membrane is a rich source of complex lipids, accounting for 24% of the total lipid intake for breast-fed infants.
From a food source, milk is rich in Sphingomyelin (SPM or SM), an important component of the myelin membrane that surrounds nerve cells, and is widely distributed in the brain and heart. SPM and sphingolipids are essential components of the central nervous system. Myelination is important for early brain development. Dihydrosphingosines (abbreviated DSPH or DHS), an important signaling molecule or second messenger (Zhang et al,2005), belongs to the class of sphingolipids and is distributed in large numbers in the central nervous system. The metabolite not only plays a role in the construction of biological membranes, but also plays a role in the signal transduction process of countless cells as a second messenger. In the brain, imbalance of different sphingolipid substances may lead to dysfunction of the nervous system and apoptosis of brain cells, eventually leading to lesions such as alzheimer's disease (Li et al, 2016). Sphinganine plays an important role in the biosynthesis and metabolism of sphingolipids. It was found that the dihydrosphingosine content is reduced in the plasma of animals suffering from alzheimer's disease. It was shown that sphinganine can be used as a biomarker for diagnosing alzheimer's disease (Li et al, 2015). Phosphatidylethanolamine (PE) has a high content in cow's milk, egg white and soybean, is the most abundant phospholipid in mammalian cell membranes, and can affect a series of cell processes and the stability and function of membrane proteins. While dietary sources of Phosphatidylserine (PS) are mainly meat and fish. The phospholipid has negative charge, is located on the inner page of a cell membrane, and can participate in cell activities including enzyme reaction, signal transduction and the like. The human brain and nerve cell membranes are rich in PS, and the PS also plays an important role in early brain development and normal function of nerves of infants. Lecithin is rich in Phosphatidylcholine (PC), and in cow milk and human milk, PC content is also high. It is a precursor of neurotransmitter acetylcholine, and has important effects on neuronal communication, memory formation and the like. Phosphatidylinositol (PI) is contained in food such as soybean and cow milk. It is a trace component of the membrane structure and can maintain the structure and the integrity of the cell membrane; it also has effects in improving immunity and increasing HDL content.
There is a need for solutions that can regulate the nervous system and improve learning and memory in the field of infant formula powders, complementary foods and nutritional supplements. Meanwhile, in the field of children, adolescents and adults over 3 years old, it is also necessary to maintain the nervous system, including the brain, to function stably and normally.
CN106106753A discloses an infant formula powder rich in various milk phospholipids, wherein the phospholipid content of the infant formula powder is increased by adding some raw materials rich in phospholipids. However, the prior art does not clearly show the technical effect of the increase of the phospholipid content.
CN101316521A discloses a polar lipid mixture comprising glycerophospholipids such as choline Phosphatidate (PC), Phosphatidylethanolamine (PE), Phosphatidylserine (PS) and inositol Phosphatidate (PI), and sphingolipids such as Sphingomyelin (SM). Most importantly, the proportion of phospholipids in the mixture is comparable to that in HMF, represented by SM > PC > PE > PS > PI or SM ═ PC > PE > PS > PI. Can be used for supporting and enhancing cognitive development in infants.
Disclosure of Invention
An object of the present invention is to provide a phospholipid composition with improved learning and memory abilities.
The invention also aims to provide the application of the phospholipid composition in preparing food with the effect of improving learning and memory abilities.
In experimental research, the inventor finds that a phospholipid composition prepared by compounding a plurality of specific phospholipids can obviously influence the chemotactic capacity of caenorhabditis elegans.
The caenorhabditis elegans as a model organism has better application prospect in preclinical research and evaluation. It has the characteristics of short life cycle (21 days), strong reproducibility and regeneration, simple and convenient operation, transparency, easy culture and the like. The genome has been completely sequenced and one quarter of the genes are homologous to the human genome. The nematode organism with gene mutation produced by editing nematode gene can be used as experimental means for gene analysis. Nematodes are not currently considered an animal in european legislation. It is widely used as an in vitro assay such as transcriptomics, proteomics, metabolomics, and the like. As a model organism, it is also often used as a step in the evaluation of raw materials.
The molecular basis for studying cognitive impairment in vivo relies on the use of animal models. Among the many alternatives, nematodes have become good models of various neurodegenerative diseases (Li and Le, 2013) and cognitive aging (Arey and Murphy, 2017). Nematodes possess 302 neurons, whose neurotransmitters and neuropeptides resemble the nervous system of mammals. All essential functions of the nematode, including development, feeding, and movement, are controlled by the nervous system. In addition, its conserved neurotransmitter biological composition and high homology to the mammalian system make nematodes a unique systemic organism for the study of neurodegenerative diseases. The end-point index method for studying the neuroprotective effects of nematodes involves studying the changes in their morphology, behavior, gene expression and neurotransmitters (nidesesh et al, 2016). In addition, nematodes show a series of behavioral characteristics and behavioral plasticity, and because individual nematodes have high gene background homology, the nematodes become a suitable animal model for studying behavioral indicators. Under some specific stress conditions, the production of excess reactive oxygen species ROS can cause nematode neuronal dysfunction. The integrity of damaged neurons can be assessed by measuring indicators of various animal motives (Wangchai et al, 2016).
Therefore, experimental studies of the present inventors have shown that a phospholipid composition prepared by compounding a plurality of specific phospholipids, including Dihydrosphingosine (DSPH), Phosphatidylcholine (PC), and Phosphatidylinositol (PI), can improve the learning and memory abilities of an organism.
In one aspect, the present invention provides a phospholipid composition comprising Dihydrosphingosine (DSPH), Phosphatidylcholine (PC), Phosphatidylinositol (PI), based on 100% total phospholipid mass in the composition:
the amount of sphinganine in the composition is 2% -10%;
the amount of phosphatidylcholine in the composition is 20% to 59%;
the amount of phosphatidylinositol in the composition is 3% to 15%.
According to a particular embodiment of the invention, the amount of sphinganine in the phospholipid composition of the invention is between 2% and 8%, preferably between 3% and 6%, based on 100% by mass of the total phospholipids in the composition.
According to a particular embodiment of the invention, the amount of phosphatidylcholine in the phospholipid composition of the invention is preferably between 25% and 55%, more preferably between 28% and 50%, based on 100% by weight of the total phospholipids in the composition.
According to a particular embodiment of the invention, the amount of phosphatidylinositol in the phospholipid composition of the invention is preferably between 12% and more preferably between 5% and 10% based on 100% by weight of the total phospholipids in the composition.
According to a particular embodiment of the invention, the ratio of the content of sphinganine to the content of phosphatidylcholine in the phospholipid composition of the invention is between 4 and 35: 100, preferably 6 to 20: 100, more preferably 6 to 18: 100.
according to a particular embodiment of the invention, the ratio of the content of sphinganine to the content of phosphatidylinositol in the phospholipid composition of the invention is 20-70: 100, preferably 35-65: 100, more preferably 41 to 58: 100.
according to a particular embodiment of the invention, the phospholipid composition of the invention comprises other Sphingomyelins (SPM) in addition to dihydrosphingosine, the total content of sphingomyelins in the composition being 15% to 26% based on 100% by mass of total phospholipids in the composition.
According to a particular embodiment of the invention, the total amount of sphingomyelin (including dihydrosphingosine), phosphatidylcholine and phosphatidylinositol in the composition is 50% or more, preferably 55% or more, more preferably 58% to 75%, based on 100% by weight of total phospholipids in the composition.
According to a particular embodiment of the invention, Phosphatidylserine (PS) may also be included in the phospholipid composition of the invention, in an amount of 6% to 15% phosphatidylinositol in the composition, based on 100% by mass of the total phospholipids in the composition.
According to a specific embodiment of the present invention, the total amount of sphingomyelin (including dihydrosphingosine), phosphatidylcholine, phosphatidylinositol and phosphatidylserine in the composition is 60% or more, preferably 65% or more, and more preferably 70% to 80%, based on 100% by weight of the total phospholipids in the composition.
According to a specific embodiment of the present invention, Phosphatidic Acid (PA) may also be included in the phospholipid composition of the present invention, and the amount of phosphatidic acid in the composition is 0.3% to 12%, preferably 0.35% to 10%, more preferably 0.4% to 8%, further preferably 0.45% to 6%, still further preferably 0.48% to 4%, and still further preferably 0.49% to 2.5%, based on 100% by mass of total phospholipids in the composition.
In the phospholipid compositions of the present invention, the source of each phospholipid component may be an animal source including, but not limited to, bovine and/or ovine milk and/or lecithin sources and the like, and/or a plant source including, but not limited to, soy sources and the like.
On the other hand, the invention also provides the application of the phospholipid composition in preparing the composition with the effect of improving the learning and/or memory ability.
According to a specific embodiment of the present invention, the composition for improving learning and/or memory is a food. For example, the food product may be a powdered milk, a liquid milk, a complementary food, a nutritional supplement, a powdered milk tea, or a milk tablet. The milk powder can be infant formula powder (infant formula food) or non-infant milk powder such as milk powder for children and milk powder for adults (including milk powder for middle-aged and elderly people). In some embodiments of the invention, the food product is an infant formula powder or an infant liquid milk.
In another aspect, the present invention further provides a food having an effect of improving learning and/or memory abilities, wherein the raw material composition of the food comprises the phospholipid composition of the present invention.
According to a particular embodiment of the invention, the phospholipid composition is applied to the food product in an amount of:
the application amount in the powder food is as follows: the mass ratio of the phospholipid composition to the total mass of the product is 0.05-20%, preferably 0.08-5%, and more preferably 0.1-2%;
the application amount in the liquid form food is as follows: the weight of the phospholipid composition is 0.0065-2.821g/100mL, preferably 0.0103-0.705g/100mL, and more preferably 0.0129-0.282g/100mL based on the total volume of the product.
The phospholipid composition and the food containing the same are beneficial to improving the learning and/or memory ability of organisms.
Drawings
Fig. 1 and 2 show the evaluation of the effect of composition 1 in the learning and memory model of nematode in example 1 of the present invention.
Fig. 3 and 4 show the evaluation of the effect of composition 2 in the online worm learning and memory model in example 2 of the present invention.
Fig. 5 and 6 show the evaluation of the effect of composition 3 in the online worm learning and memory model in example 3 of the present invention.
Fig. 7 and 8 show the evaluation of the effect of composition 4 in the nematode learning and memory model in example 4 of the present invention.
Fig. 9 and 10 show the evaluation of the effect of composition 5 in the nematode learning and memory model in example 5 of the present invention.
Fig. 11 and 12 show the evaluation of the effect of composition 6 in the nematode learning and memory model in example 6 of the present invention.
FIG. 13 shows the percentage increase in the learning coefficient of the organism after 45 minutes exposure compared to the control for the different phospholipid compositions of example 7.
FIG. 14 shows representative pictures of the chemotaxis assay for each of the compositions of example 8.
Detailed Description
For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description of the technical aspects of the present invention with reference to specific examples, which are intended to illustrate the present invention and not to limit the scope of the present invention.
Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art. The content of each phospholipid component was measured by a method conventional in the art in the examples. The operating conditions not specified in detail in the examples were carried out according to the usual procedures in the art.
In addition, in order to avoid repetition, general procedures required to be followed in the experiments in each example, such as preparation of a culture medium, and the like, are listed below.
Different phospholipid raw materials (comprising solid and/or liquid PC raw materials, milk phospholipid raw materials, PS raw materials and the like) are selected to prepare each phospholipid composition sample of the invention, wherein the content of each phospholipid component is shown in the table 1, and the rest components are small amount of protein, fat and the like (the components mainly play roles in constructing tissues and supplying energy in organisms). Dissolving the raw materials with distilled water to obtain raw solution. The material was then added to the agar surface and Nematode Growth Medium (NGM) was added. Considering the difference of the total phospholipid content of each composition raw material, each composition raw material is prepared to ensure that the final dosage of each group added into the culture medium in the experiment keeps the total phospholipid content consistent and is 20 mu g.
TABLE 1 phospholipid compositions tested and the ratio ranges of the monomers
Figure BDA0003385679890000071
Model for experimental learning and memory of nematode
Nematode chemotaxis assays were modeled with reference to published studies (Stein and Murphy, 2014). Wild type N2 nematodes were first cultured on NGM plates and were of uniform age, and E.coli OP50 was added to the medium as feed for the nematodes. Various combinations of phospholipid materials (including soy or lecithin, milk phospholipids enriched in phosphatidylserine, etc.) are added to the NGM plates. The total phospholipid content of these different combinations of phospholipid material was consistent, allowing for lateral comparisons of the different results.
The nematodes will first be in a starvation environment and will then be exposed to a butanone environment to train them for a positive association of food with butanone (the trained nematode group). To assess how phospholipids affect nematode learning ability (rate of nematode learning), different butanone exposure times (e.g. 0, 15, 30, 45 and 60 minutes) will be studied and corresponding chemotaxis tests performed, and learning index under each condition is recorded (Kauffman et al, 2010).
Young adult nematodes, which may be untrained or butanone-trained, will be collected, washed with buffer, and placed in chemotaxis plates according to established methods (Kauffman et al, 2011; Margie et al, 2013). The plate was divided into four quadrants and would contain either the chemokine (test quadrant) or the control solution (control quadrant). The chemotactic Index Chemotaxis Index, CI ═ number of attracted nematodes (number of attracted nematodes accumulating in test quadrant-number in control quadrant)/total number of nematodes, was obtained in untrained or trained nematodes. Thereafter, the Learning Index, LI, is calculated, and LI is trained-untrained CI. The learning index of the control group without phospholipid added will then be compared to the learning index of the phospholipid test group at different time points. Representative chemotaxis test plate pictures are also shown in the results.
Instruments and materials
Stereo microscope (Motic SMZ-168)
Microorganism safety console (Telstar Bio-II-A)
Incubator (Memmert)
Single channel liquid-moving device (Gilson)
Vortex mixing instrument
Platinum wire
Alcohol burner
NGM Medium (3g/L sodium chloride, 17g/L agar, 2.5g/L peptone, 1mM magnesium sulfate, 25mM potassium phosphate buffer, 5. mu.g/mL cholesterol)
Culture plate (55mm X15 mm)
Escherichia coli OP50
Distilled water
Filter head
Conical centrifuge tube
1.5mL small test tube
Butanone (Sigma-Aldrich, reference number W217012)
96% ethanol (reference number ET0003005P)
Sodium azide (Sigma-Aldrich, reference number S2002)
Time-meter
Data analysis
To optimize the experimental workload, the test concentrations were first specified. For each dose and sample, two independent experiments will be performed simultaneously and subjected to mathematical statistical analysis. Statistical analysis was performed with GraphPad Prism 9. At the same time point, the differences between the control and experimental groups were statistically analyzed using t-test. The significance of the differences between the groups is indicated by an asterisk, P <0.05, P <0.01, P < 0.001. The letter NS indicates that the difference is not significant.
Example 1: composition 1 evaluation of Effect in an nematode learning and memory model
The preparation steps and specific experimental methods before the experiment are described in the preceding paragraphs. The learning capacity of the test substances for nematodes was found to be slightly improved at the 30 min butanone exposure time point (P <0.05), with a significant improvement at 60 min, while at 45 min a very good effect was produced (P <0.01) on significantly increasing chemotactic and learning coefficients (fig. 1, fig. 2).
Example 2: composition 2 evaluation of Effect in an nematode learning and memory model
The preparation steps and specific experimental methods before the experiment are described in the preceding paragraphs. Based on previous experimental data, the same total phospholipid content, i.e. 20 μ g, was set for all other subsequent phospholipid compositions tested.
Nematodes co-cultured with composition 2 showed significant increases in chemotactic index after 45 min (P <0.001) and 60 min (P <0.01) exposure compared to the control (figure 3). Similarly, there was also a significant improvement in the learning coefficient at these two time points, where P <0.01 at 45 minutes and P <0.05 at 60 minutes (fig. 4).
Example 3: composition 3 evaluation of Effect in an nematode learning and memory model
Nematodes co-cultured with composition 3 showed a significant increase in chemotactic index after 45 min exposure (P <0.05) compared to the control (figure 5). Similarly, the learning coefficient at this time point was also significantly improved (P <0.05), as shown in fig. 6. There was a slight, but not significant, trend toward a slight increase in chemotactic and learning coefficients after incubation for 30 minutes.
Example 4: composition 4 evaluation of Effect in an nematode learning and memory model
Figures 7 and 8 show the performance of the learning and cognition models after co-culture of nematodes with composition 4. The results show the highest increase in the degration coefficient after 45 minutes of butanone exposure, with a more pronounced effect on the increase in chemotactic coefficient at 60 minutes. And the improvement of the learning coefficient is reflected in the 45 th minute, and the difference between the experimental group and the control group is significant (P < 0.05).
Example 5: composition 5 Effect assessment in an nematode learning and memory model
Experiments showed that composition 5 did not have any significant improvement in both chemotactic and learning coefficients at this tested concentration at different butanone exposure times (fig. 9, 10).
Example 6: composition 6 evaluation of Effect in an nematode learning and memory model
Composition 6 showed significant increases in both chemotactic and learning coefficients (P <0.01) after 45 min of butanone exposure in co-incubation with nematodes, and slightly, but not significantly, after 60 min of exposure, as shown in fig. 11 and 12.
Example 7: comparison of the Effect of different phospholipid compositions on improving learning ability
To better compare the effects of different tested phospholipid compositions, fold change calculations (learning coefficient/control group learning coefficient for experimental group) were performed after butanone exposure for 45 minutes (time point where the highest fold change, i.e. the most significant effect, was observed) based on the values of the learning coefficients (table 2). In addition, the percent increase in learning coefficient compared to the control for each group after 45 minutes exposure is shown in fig. 13. In all groups tested, composition 3 exhibited the most significant improvement in learning and memory in the nematode model (34.7% improvement in learning coefficient over the NGM control by a factor of 1.34), followed by composition 1, 30% improvement in learning coefficient over the NGM control by a factor of 1.31. The composition 2 and the composition 4 improve the learning and memory abilities as well, the learning coefficients are respectively increased by 20.7 percent and 25.9 percent, and the improvement multiples are respectively 1.22 and 1.25. As for composition 5, there was no boosting effect. Composition 6, although statistically significant, had the least effect on learning ability improvement.
TABLE 2
Phospholipid composition group Percentage of learning coefficient improvement over control Multiple of change Whether is significant or not
Composition 1 30.0 1.31 Is that
Composition 2 20.7 1.22 Is that
Composition 3 34.7 1.34 Is that
Composition 4 25.9 1.25 Is that
Composition 5 -3.1 0.98 Whether or not
Composition 6 8.2 1.06 Is that
The multiple of 1 indicates no change to the learning coefficient;
a multiple >1 indicates a positive boosting effect on the learning coefficient;
a factor <1 indicates a reduced negative effect on the learning coefficient.
Example 8: representative photographs of chemotaxis assays for each phospholipid composition
Figure 14 shows representative pictures of chemotaxis assays for each composition. Where the plate was divided into four quadrants, T is the quadrant containing the chemotactic agent butanone and C is the control solution quadrant. The left chemotaxis test plate is the NGM control plate, while the right chemotaxis test plate is the plate co-cultured with the nematode and phospholipid composition, composition 1 (panel a), composition 2 (panel B), composition 3 (panel C), composition 4 (panel D), composition 5 (panel E), composition 6 (panel F). Different effects of the compositions on the increase in chemotactic capacity of nematodes can be observed.
Example 9: infant formula containing phospholipid composition (1000 kg prepared)
The starting materials for the infant formula containing the phospholipid composition of this example: 1000 kg of raw milk, 320 kg of lactose, 30 kg of whey protein powder WPC 80%, 90175 kg of desalted whey powder D, 40 kg of corn oil, 50 kg of soybean oil, 140 kg of OPO structural fat, 27 kg of alpha-whey protein powder, 9 kg of beta-casein powder, 1 kg of anhydrous cream, 17 kg of fructo-oligosaccharide powder and 40 kg of galacto-oligosaccharide syrup, 10 kg of any one of the phospholipid compositions 1 to 4, 16 kg of compound nutrient, 12 kg of DHA, 22 kg of ARA and 0.2 kg of bifidobacterium.
The compound nutrient comprises about 2.5 kg of compound vitamin nutrient package, about 0.75 kg of choline chloride nutrient package, about 6 kg of calcium powder nutrient package, about 1 kg of mineral nutrient package, about 1.5 kg of magnesium chloride nutrient package and about 2 kg of potassium chloride nutrient package, and the base material of each nutrient package is lactose.
The preparation process of the infant formula containing the phospholipid composition of this example is as follows:
1) milk rough filtration: after coarse filtration and degassing in a balance cylinder, the milk is preheated by a plate heat exchanger, and impurities are separated by a separator.
2) Homogenizing and sterilizing milk: one part of the raw milk without impurities enters a homogenizer for homogenization, the other part of the raw milk is inhomogeneous, and the homogenized raw milk are mixed and enter a sterilization system for sterilization.
3) Adding powder: various powder raw materials are metered according to the formula, uniformly added into a powder preparation tank through an air conveying system, and sucked into a vacuum mixing tank through a vacuum system.
4) Dissolving and oil blending: the grease and phospholipid composition specified in the formula is placed into an oil dissolving chamber according to the formula requirement, the temperature of the oil dissolving chamber is kept between 50 and 90 ℃, after the oil is dissolved, the oil dissolving chamber is filled into a mixed oil storage tank, and the mixed oil is filled into a material mixing tank through an oil pump according to the formula requirement.
5) Dissolving and adding nutrients: respectively dissolving nutrient bags such as calcium powder, vitamins, minerals, etc. with purified water, and sequentially adding into a mixing tank to obtain mixed feed liquid.
6) And (3) filtering: filtering the mixed feed liquid by a filter screen to remove physical impurities possibly brought in the raw materials.
7) Homogenizing: homogenizing the mixed material liquid by a homogenizer, mechanically processing the fat balls, and dispersing the fat balls into uniform fat balls.
8) Cooling and storing: and (3) feeding the homogenized material liquid into a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storage cylinder, entering the next procedure within 6 hours, and starting the stirrer according to the set requirement.
9) Concentration and sterilization: double-effect concentration is adopted during production, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharge concentration was 50% dry matter.
10) Storing concentrated milk, preheating, filtering and spray drying: the concentrated milk is temporarily stored in a concentrated milk balancing tank. Preheating to 60 deg.C with a scraper preheater, filtering the preheated material with a filter with 1mm pore diameter, pumping into a drying tower with a high pressure pump, spray drying, and agglomerating fine powder at the tower top or fluidized bed as required. Air inlet temperature: 180 ℃, the exhaust temperature is 86 ℃, the pressure of the high-pressure pump is 200bar, and the negative pressure of the tower is about-4 mba.
11) Drying and cooling the fluidized bed: and (3) drying the powder from the drying tower for the second time by the fluidized bed (the first stage), and cooling to 30 ℃ by the fluidized bed (the second stage) to obtain the main milk powder material.
12) Subpackaging: and (3) weighing DHA, ARA or bifidobacterium, sealing bags and subpackaging by powder-making workshop personnel according to the formula requirements.
13) Dry mixing: and uniformly mixing the weighed DHA, ARA or bifidobacteria and the milk powder main material in a dry mixer.
14) Powder sieving: the granularity of the milk powder is uniform through the vibrating screen, and the powder residue is discarded.
15) Powder discharging: and (4) receiving the powder by using a sterilized powder collecting box, and conveying the powder to a powder feeding room from a powder discharging room.
16) Powdering: pouring the milk powder into a powder storage tank on a large and small packaging machine according to the packaging requirements.
17) Packaging: 400 g of the mixture is packaged by an automatic packaging machine in a nitrogen-filled mode. The oxygen content is lower than 1% when charging nitrogen. The oxygen content of the 900 g iron can in the automatic nitrogen-filled package is lower than 5 percent.
18) Boxing: and (4) filling the packaged small bags into a paper box, adding a powder spoon, and sealing by using a box sealing machine.
19) And (4) inspecting a finished product: and sampling and inspecting the packaged product according to an inspection plan.
20) And (4) warehousing and storing: and warehousing and storing the qualified product at normal temperature with the humidity less than or equal to 65 percent.
Through detection, all indexes of the milk powder respectively added with any one of the phospholipid compositions 1-4 prepared by the method meet the relevant standard requirements of infant formula food.

Claims (14)

1. A phospholipid composition comprising Dihydrosphingosine (DSPH), Phosphatidylcholine (PC), Phosphatidylinositol (PI), based on 100% total phospholipid mass in the composition:
the amount of sphinganine in the composition is 2% -10%;
the amount of phosphatidylcholine in the composition is 20% to 59%;
the amount of phosphatidylinositol in the composition is 3% to 15%.
2. A phospholipid composition according to claim 1 wherein the amount of sphinganine in the composition is between 2% and 8%, preferably between 3% and 6%, based on 100% by mass of total phospholipids in the composition; and/or
The content of phosphatidylcholine in the composition is 25% -55%, preferably 28-50%; and/or
The amount of phosphatidylinositol in the composition is 4% to 12%, more preferably 5% to 10%.
3. The phospholipid composition of claim 1, wherein the ratio of the content of sphinganine to the content of phosphatidylcholine is between 4-35: 100, preferably 6 to 20: 100, more preferably 6 to 18: 100.
4. a phospholipid composition according to claim 1 or claim 3 wherein the ratio of sphinganine content to phosphatidylinositol content is from 20 to 70: 100, preferably 35-65: 100, more preferably 41 to 58: 100.
5. a phospholipid composition according to claim 1 which comprises a Sphingomyelin (SPM) in addition to sphinganine, the total content of SPM in the composition being from 15% to 26% based on 100% by mass of total phospholipids in the composition.
6. A phospholipid composition according to claim 1 or claim 5 which further comprises Phosphatidylserine (PS) in an amount of from 6% to 15% phosphatidylinositol based on 100% total phospholipid mass in the composition.
7. A phospholipid composition according to claim 1, which further comprises Phosphatidic Acid (PA) in an amount of from 0.3% to 12%, preferably from 0.49% to 2.5%, based on 100% total phospholipids in the composition.
8. A phospholipid composition according to any one of claims 1 to 7 wherein the total amount of sphingomyelin, phosphatidylcholine and phosphatidylinositol in the composition is 50% or more, preferably 55% or more, more preferably 58% to 75% by weight of the total phospholipids in the composition as 100%; and/or
The total content of sphingomyelin, phosphatidylcholine, phosphatidylinositol and phosphatidylserine in the composition is more than 60%, preferably more than 65%, more preferably 70% -80%.
9. Use of a phospholipid composition according to any one of claims 1 to 8 in the preparation of a composition for enhancing learning and/or memory.
10. The use according to claim 9, wherein the composition having the effect of improving learning and/or memory is a food.
11. The use according to claim 10, wherein the food product is a powdered milk, liquid milk, a complementary food or a nutritional supplement;
preferably, the food is infant formula powder or infant liquid milk.
12. A food having an effect of improving learning and/or memory ability, which comprises the phospholipid composition according to any one of claims 1 to 8 as a raw material.
13. A food product as claimed in claim 12 wherein the phospholipid composition of any one of claims 1 to 8 is applied to the food product in an amount of:
the application amount in the powder food is as follows: the mass ratio of the phospholipid composition to the total mass of the product is 0.05-20%, preferably 0.08-5%, and more preferably 0.1-2%;
the application amount in the liquid form food is as follows: the weight of the phospholipid composition is 0.0065-2.821g/100mL, preferably 0.0103-0.705g/100mL, and more preferably 0.0129-0.282g/100mL based on the total volume of the product.
14. A food product according to claim 12 or 13 which is a milk powder, liquid milk, a complementary food or a nutritional supplement;
preferably, the food is infant formula powder or infant liquid milk.
CN202111453109.7A 2021-11-30 2021-11-30 Phospholipid composition capable of improving learning and memory ability and application thereof Pending CN114145347A (en)

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Citations (4)

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Publication number Priority date Publication date Assignee Title
WO2008081934A1 (en) * 2006-12-28 2008-07-10 Meiji Dairies Corporation Agent for facilitating the development of brain in infant comprising milk-derived phospholipid and food composition comprising the same
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CN106912614A (en) * 2017-03-10 2017-07-04 海普诺凯营养品有限公司 A kind of humanized formula milk and preparation method thereof
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WO2008081934A1 (en) * 2006-12-28 2008-07-10 Meiji Dairies Corporation Agent for facilitating the development of brain in infant comprising milk-derived phospholipid and food composition comprising the same
WO2015086169A1 (en) * 2013-12-12 2015-06-18 Nestec S.A. Synthetic milk compositions comprising polyunsaturated fatty acids (pufas) for promoting the healthy establishment of cognitive function in male and female infant and children
CN108366603A (en) * 2015-12-14 2018-08-03 雀巢产品技术援助有限公司 The alimentation composition and infant formula for promoting brain myelin sheath to be formed
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