CN115813882A - A method for enhancing absorption of vitamins and minerals in intestinal tract - Google Patents

Abstract

The invention discloses a method for enhancing the absorption of vitamins and mineral substances in intestinal tracts, which comprises the steps of complexing plant polyphenol and mineral substances to form a nano-network material, and combining the nano-network material with vitamin molecules to form a polyphenol-based vitamin compound; the polyphenol-based vitamin complex has long retention time in intestinal tract, and can promote the absorption of vitamins and minerals in intestinal tract. The mineral is Zn ²⁺ Or Fe ²⁺ . The plant polyphenol is one of tannic acid, epigallocatechin gallate, rutin, and anthocyanidin. The vitamins are selected from one or more of oil soluble vitamin A, vitamin D, and vitamin E, or one or more of water soluble vitamin C, vitamin B2, vitamin B12, calcium pantothenate, and folic acid. The invention utilizes plant polyphenol and metal ions to modify vitamin molecules, constructs an effective vitamin intestinal delivery system, and utilizes the interaction of the plant polyphenol and intestinal wall cell surface protein to enhance the absorption of vitamins and minerals.

Description

A method for enhancing absorption of vitamins and minerals in intestinal tract

Technical Field

The invention relates to the technical field of vitamin delivery, in particular to a method for enhancing the absorption of vitamins and minerals in intestinal tracts.

Background

Vitamins are a kind of nutrient substances necessary for maintaining body health, are low molecular organic compounds, and have very important effects on metabolism, growth, development and health of human and animal organisms. Minerals are a class of inorganic salts that, although less than 5% of the body weight is present in the human body, do not provide energy, but also play an important role in the physiological actions of human tissues.

Vitamins are a class of organic substances that are essential to the maintenance of human life and can only be obtained from food, and are classified into oil-soluble and water-soluble. Common oil-soluble vitamins are Vitamin A (VA), vitamin D (VD) and Vitamin E (VE). The water-soluble vitamins include Vitamin C (VC), vitamin B group such as B2, B5 (calcium pantothenate), B9 (folic acid), and B12. Supplements and products for vitamins and minerals in the market are of various brands and varieties. Under the call of modern nutrition advocating 'diversification', the composite supplement is popular with vitamin-nutrient mineral nutritional supplements. However, the prior product has serious technical homogeneity, is mainly subjected to iterative updating on formulas, namely the types and the dosages of vitamins and minerals, has the problems of insufficient additive dosage or excessive dosage, and does not fundamentally solve the problems of absorption and utilization rate of the vitamins. Apart from a balanced match, absorption is the key to the use of vitamins and minerals.

Oil-soluble vitamins can be analogized to common hydrophobic drugs. The delivery of hydrophobic drugs, the manner of preparation of oral formulations, can be applied to oil-soluble vitamin applications. It is challenging to deliver water-soluble vitamins. Water-soluble vitamins are generally administered in the form of chewable tablets, oral tablets, or granules. Such formulations require rapid disintegration of the dosage form when exposed to water to render the vitamin as water soluble as possible. The vitamins dissolved in water are small molecules which are free in the solution, and due to good reducibility, the vitamins are easily oxidized and decomposed by other substances and cannot be fully absorbed and utilized. The polarity of hydrophobic drugs is not suitable for the processing of water-soluble drugs. Therefore, improving the delivery and absorption efficiency of water-soluble vitamin molecules is key to effective vitamin supplementation. In addition, the existing products are all prepared into tablets or granules by blending different vitamins, and the antagonism among different vitamins is not considered. For example, large amounts of vitamin E consume vitamin a stores in the human body; vitamin C and vitamin B are easy to generate oxidation-reduction reaction, so that the vitamins lose efficacy; vitamin C and folic acid should be used off peak; the presence of large amounts of vitamin E can affect the absorption of vitamin K.

There are reports in the literature of modifications to fat-soluble and water-soluble vitamins, respectively. For example, patents CN113727706a and CN113747886a, select some biopolymers, such as alginate, chitosan, pectin, cyclodextrin, etc. These substances protect vitamins, are stable in the gastric acid environment and decompose in the intestinal environment. These materials and techniques enable the delivery of vitamins in the intestinal tract. However, these materials have not been shown to aid in the absorption of vitamins in the gut; only a sufficient absorption, vitamin and mineral supplement is of interest.

For the supplement of the minerals iron and zinc, the metal salts such as ferrous sulfate or the oxides such as zinc oxide are mainly used in the market at present. The metal salt and the oxide are fully dissolved in gastric juice to form an ion form and then enter the intestinal tract for absorption. However, this method is not efficient in mineral absorption. In gastric juice and intestinal juice, various digestive enzymes and substances produced by metabolism of other nutrients are usually present, and the substances are easy to react with the metal ions to regenerate water-insoluble metal salts, such as oxalates, so that the metal ions are rapidly precipitated and directly discharged out of the body without being absorbed. Therefore, the absorption of metal ions is effectively improved, and the mineral supplement is very important.

The gastrointestinal tract is an important site for vitamin absorption, and both vitamins and minerals are absorbed into the blood through a unique transport mechanism of intestinal wall cells. Due to the acceleration of modern life rhythm, irregular diet, diseases, treatment medicines and dysbacteriosis, intestinal tract reaction can be caused, the normal function of epithelial cells on the intestinal wall is influenced, and the absorption of vitamins is seriously hindered. Currently there is no effective way to specifically target the enhanced intestinal absorption of vitamins and minerals.

Disclosure of Invention

In order to enhance the absorption of vitamins and minerals in the gastrointestinal tract, the present invention provides a method for enhancing the absorption of vitamins and minerals in the gastrointestinal tract.

The method for enhancing the absorption of vitamins and minerals in the gastrointestinal tract provided by the invention utilizes plant Polyphenol and minerals to complex to form a nano-network Material (MPN), the nano-network material is combined with vitamin molecules, and a Polyphenol-based vitamin complex (PVC) is formed by coating or agglomeration adsorption. The polyphenol-based vitamin complex has long retention time in the gastrointestinal tract, and can promote the absorption of vitamins and minerals in the gastrointestinal tract.

The plant polyphenol is one of tannin, epigallocatechin gallate, rutin, and anthocyanins.

The mineral is Zn ion ²⁺ Or ferrous ion Fe ²⁺ 。

The method is suitable for oil-soluble vitamins and water-soluble vitamins. The oil soluble vitamin may be one or more of vitamin A, vitamin D, and vitamin E. The water soluble vitamin can be one or more of vitamin C, vitamin B2, vitamin B12, calcium pantothenate, and folic acid.

Aiming at oil-soluble vitamins, the preparation method of the polyphenol-based vitamin complex comprises the following steps:

(1) Dissolving vitamins in soybean oil to prepare an oil agent, and adding the oil agent into a phosphate buffer solution for ultrasonic treatment to form a vitamin emulsion;

(2) Dissolving plant polyphenol in ultrapure water to form a polyphenol water solution; and will contain Zn ²⁺ Or Fe ²⁺ Dissolving the soluble salt in another part of ultrapure water to obtain a mineral water solution;

(3) Adding vitamin emulsion into polyphenol water solution, performing ultrasonic treatment, adding mineral water solution, mixing, centrifuging, and collecting upper emulsion to obtain polyphenol-based vitamin complex.

Aiming at water-soluble vitamins, the preparation method of the polyphenol-based vitamin complex comprises the following steps:

(1) Adding vitamins into a phosphate buffer solution to form a vitamin solution;

(2) Dissolving plant polyphenol in ultrapure water to form a polyphenol water solution; and will containZn ²⁺ Or Fe ²⁺ Dissolving the soluble salt in another part of ultrapure water to obtain a mineral water solution;

(3) Adding a vitamin solution into a polyphenol water solution, and stirring at room temperature to ensure that the vitamin is fully contacted with the plant polyphenol to form nanoparticles by agglomeration; then adding mineral water solution, stirring well, dialyzing the obtained solution under low concentration phosphate buffer solution for 2 days to remove impurities, and cutting off 3.0kDa of molecular weight; and freeze-drying the obtained polyphenol-based vitamin compound solution to prepare polyphenol-based vitamin compound dry powder.

In the method, the plant polyphenol is a plant secondary metabolite, the molecular structure contains a large amount of pyrogallol and catechol groups, and the plant polyphenol has the characteristic of strong complexation with metal ions. The network structure Material (MPN) composed of metal ions and plant polyphenol is a nano delivery platform with great potential, the preparation process is simple and rapid, and the network structure material has the characteristic of modularized raw material selection. It contains multiple edible plant polyphenols and multiple metal ions (mineral: ferrous ion Fe) ²⁺ Zinc ion Zn ²⁺ Etc.) may be selected. Plant polyphenols, also known as plant tannins, have long been used as tanning agents in the tanning industry. The leather tanning process is a process for crosslinking phenolic hydroxyl of plant polyphenol with skin collagen. The collagen is a kind of protein, so the plant polyphenol and the protein can be strongly combined, namely the plant polyphenol and the protein of the intestinal wall cell can be strongly combined through hydrogen bonds. The phenolic hydroxyl and aromatic group of the plant polyphenol can interact with different functional groups in various modes such as hydrogen bond, chelation, amphipathy, pi-pi superposition and the like. Therefore, MPN formed by complexing plant polyphenol and metal ions can be combined with vitamin molecules, and on the other hand, the metal ions in the structure can be simultaneously used as a mineral source for nutrition supplement. Based on the method, the plant polyphenol and the metal ions are used for modifying vitamin molecules, and the interaction of the plant polyphenol and the surface protein of intestinal wall cells is used for enhancing the absorption of vitamins and minerals.

Compared with the prior art, the invention has the advantages that:

(1) The modification of vitamins by the metal-polyphenol network material MPN can prolong the retention time of the vitamins in intestinal tracts.

(2) Modification of water-soluble vitamins by the metal-polyphenol network material MPN can form vitamin-containing nanoparticles, and can help absorption of vitamins compared with water-soluble small molecules. Meanwhile, the metal ions can be used as the supplement of minerals, so that the concentration of the metal ions in blood is increased, and the absorption of the mineral metal ions is promoted. Pharmacokinetics and biodistribution studies show that PVC can significantly prolong the retention and absorption time of vitamins in intestinal tracts. Blood concentration tests also show that the content of vitamins and minerals in blood is increased.

(3) The method for constructing the vitamin delivery carrier is a process of self-assembly of plant polyphenol and metal ions on the surface of the vitamin, and is convenient and rapid to operate.

Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.

Drawings

PVC prepared in FIG. 1, examples 1 and 2 _E Oil-soluble vitamin model (a) and PVC _C Transmission electron micrograph of water-soluble vitamin model (b).

FIG. 2 is a UV-visible diagram of a nutritional model for PVC water-soluble (a-e) and oil-soluble (f-g) vitamins.

FIG. 3, measurement of Zeta potential of PVC water-soluble (a) and oil-soluble (b) vitamin nutrition models.

FIG. 4 is a graph showing the particle size before and after vitamin modification.

Fig. 5, an anatomical diagram of the jejunum and ileum.

FIG. 6, pharmacokinetic analysis chart. In the figure: (a) -oil soluble vitamins, (b) -water soluble vitamins.

FIG. 7 comparison of iron content in blood 3h after feeding rat multivitamin mineral supplement and PVC multivitamin mineral supplement.

Detailed Description

The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.

The plant polyphenol used in the following examples is epigallocatechin gallate (EGCG), tannic Acid (TA), rutin or anthocyanins. Epigallocatechin gallate is a typical plant polyphenol, is the main component of green tea polyphenol, and accounts for 40-60%. More and more studies have shown that EGCG has multiple biological functions, such as anti-inflammatory, antioxidant, immunomodulating, antitumor and modulating intestinal flora. Tannin is extracted from Galla chinensis, is hydrolyzed tannin, and has astringent and astringent effects. The molecular weight is about 1700, and the structure contains a plurality of pyrogallol groups. In medicine, gallic acid is a hemostatic astringent and has antibacterial effect. It has good reducibility, and can be used as antiseptic and antioxidant.

Example 1

Preparation of a polyphenol-based vitamin complex PVC:

mixing oil soluble vitamin E (or vitamin D, vitamin A) with EGCG and Fe ²⁺ Preparation of PVC by mixing _E (or PVC) _D 、PVC _A ) Oil soluble vitamin nutrition model.

Dissolving vitamin E in soybean oil (with an equal volume ratio v: v = 1:1) to prepare an oil agent; treating the oil solution with KQ-100DE ultrasonic cleaner (Kunshan ultrasonic apparatus Co., ltd.) in phosphate buffer solution (0.05mmol, pH7.2) by ultrasonic wave (100W) for 180s to form VE emulsion; wherein the concentration of vitamin E is 5mgmL ^-1 . FeSO (ferric oxide) is added ₄ ·7H ₂ O(2.8mgmL ^-1 ) And EGCG (1 mgmL) ^-1 ) Dissolved in two portions of ultrapure water, respectively. Adding 1mLVE emulsion into 0.5mLEGCG aqueous solution, and performing ultrasonic treatment at 100W for 120s; then 27. Mu.L of Fe was added ²⁺ Solution of Fe ²⁺ The molar ratio to EGCG is 1:4; the total volume was made up to 2ml using PBS. Centrifuging the obtained solution (5000 r/min) for 5 min, and collecting the upper emulsion to obtain PVC _E An emulsion.

The PVC oil-soluble vitamins marked by fluorescence are used for animal experiments to verify the vitamin absorption kinetics. Will fluoresceSubstance coumarin C6 (0.2mL, 1mgmL) ^-1 ) And 1mL vitamin A, D, E dissolved in 10mL dichloromethane stirring overnight; the solvent was removed using a rotary evaporator to give a fluorescently labeled vitamin A, D, E mixture. The PVC is then built up as described above _ADE Oil-soluble vitamins.

Example 2

Preparation of a polyphenol-based vitamin complex PVC:

mixing water soluble VC (or VB2, VB5, VB9 and VB 12) with EGCG and Fe ²⁺ Preparation of PVC by mixing _C A water-soluble vitamin nutrition model.

VC(5mg mL ^-1 ) Prepared with phosphate buffer (0.05mmol, pH 7.2) to obtain VC solution. Respectively reacting FeSO ₄ ·7H ₂ O(2.8mg mL ^-1 ) And EGCG (1 mg mL) ^-1 ) Dissolving in two portions of ultrapure water to obtain two aqueous solutions. The 1ml of lvc solution was added to the 0.5ml of gcg aqueous solution and stirred at room temperature for 30min to allow the vitamins to fully contact EGCG to agglomerate and form nanoparticles. Then 27. Mu.L of Fe was added ²⁺ Solution of Fe ²⁺ The molar ratio to EGCG was 1:4.Fe ²⁺ The agglomerated particles can be stabilized. The total volume was made up to 4ml using PBS. The resulting solution was dialyzed against a low concentration phosphate buffer (0.01mmol, pH 7.2) for 2 days to remove impurities (molecular weight cut-off 3.0 kDa). Then, PVC _C The solution was prepared as a dry powder using a freeze-drying method.

The fluorescence labeled EGCG is used for constructing a PVC water-soluble vitamin nutrient substance model, and the fluorescein FITC labeled EGCG (or TA) is used. EGCG (or TA) (50 mg) and FITC (1 mg) were dissolved in 9mL of physiological saline and 1mL of phosphate buffer (0.05mmol, pH 9.0). FITC-labeled EGCG (TA) was dialyzed against PBS (0.01M, pH 7.2) in the dark for 2 days (molecular weight cut-off 3.0 kDa) and lyophilized to obtain. Then, the PVC water-soluble vitamin is constructed according to the method.

Comparative example

Preparation of vitamin mineral complex for control group:

mixing oil soluble vitamin emulsion (10 mgmL) ^-1 ) With a water-soluble vitamin solution (10 mgmL) ^-1 ) Mixing the mixture in a volume ratio of 1:1, adding FeSO ₄ ·7H ₂ O(2.8mgmL ^-1 ) 54 μ L of the solution. Stirring at room temperature for 1h, and lyophilizing to obtain powder, i.e. constructing common vitamin mineral complex of control group.

The properties are characterized as follows:

(1) Observation of PVC with Transmission Electron microscopy (TEM, thermoFisher scientific Inc., USA) _E 、PVC _C Vitamin nutrition model. As shown in FIG. 1, the negative-stained TEM photograph shows PVC _E The oil-soluble vitamin model exhibits a core-shell structure, whereas PVC _C The water-soluble vitamin model is irregular-shaped particles, and the irregular shape shows that the polyphenol-vitamin-metal particles form the composite nano particles.

(2) As shown in FIG. 2, the UV-visible spectrum (UV-Vis, thermoNanoDrop2000, USA) shows passage through EGCG-Fe ²⁺ The modification of the complex, the absorption peak of water-soluble and oil-soluble vitamins appears around 330nm, which proves that EGCG-Fe ²⁺ The complex forms a thin PVC shell on the surface of the vitamin.

(3) The results of the Zeta-potentiometer analysis are shown in FIG. 3, which shows the formation of a PVC skin on the vitamin surface. The Zeta potential values of the water-soluble vitamins are respectively reduced from-3.9 (VC), -14.6 (VB 2), -12.3 (VB 5), -12.4 (VB 9) and-5.2 (VB 12) to-9.2 (VC), -24.8 (VB 2), -18.4 (VB 5), -28.1 (VB 9) and-18.7 (VB 12); the Zeta potential value of the oil-soluble vitamin is reduced from-14.6 (VA), -15.8 (VD 3), -12.3 (VE) to-20.1 (VA), -24.8 (VD 3), -21.4 (VE). Through EGCG-Fe ²⁺ The modified nutrient, zeta potential, has shifted negatively in value because the polyphenol-metal complex has a more negative potential, changing the charge properties of the vitamin particle surface.

(4) Dynamic Light Scattering (DLS) test. FIG. 4 is a graph showing the particle size before and after vitamin modification. The figure (a) is oil-soluble vitamin A, (b) is vitamin D3, (c) is vitamin E, and (D) the water-soluble vitamin is EGCG-Fe ²⁺ And (4) characterizing the particle size before and after modification. As shown in the figure, the oil-soluble vitamin can be dispersed in water to form nano-scale colloidal particles with the particle sizes of 243 (VA), 239 (VD 3) and 242 (VE) nm respectively. Through EGCG-Fe ²⁺ After the modification, the particle size is not obviously changed, which indicates that EGCG-Fe ²⁺ A thin shell layer is formed on the surface of the oil-soluble vitamin. The water solution vitamin has no particle size in the solution, and EGCG-Fe is added ²⁺ Then, nanoparticles are formed, the particle size is 100-400nm, and EGCG-Fe is shown ²⁺ Reacts with water-soluble vitamins and is agglomerated to form particles.

(5) Animal experiments

To investigate the extended residence and absorption time of PVC vitamins, 2mL concentration 2mgmL ^-1 PVC of _ADE The oil-soluble vitamins and the PVC water-soluble vitamins are mixed into a solution, the PVC mixed vitamins are prepared to be orally taken by healthy rats, and the rats are subjected to intragastric administration by taking the common mixed vitamins as a control group. Blood is drawn for 0-8h respectively for blood vitamin content detection. After 24h the rats were sacrificed and the gastrointestinal conditions were dissected. A part of rats are sacrificed at 2h for dissection, and 2-3 cm jejunal tissues are cut and fixed by 4% paraformaldehyde for morphology observation.

To evaluate the effect of PVC intake in vivo and taking into account the metabolism of vitamins, some rats were sacrificed after 2h of gavage, the gastrointestinal tract was dissected and the tissue morphology of the jejunum and ileum was observed at 2h (FIG. 5). The difference between PVC vitamins and ordinary vitamins in jejunum and ileum was significant at 2 h. The photographs of the dissection show that the PVC vitamins adhere to the mucosal side of the jejunum and ileum after 2 hours (the dark material in the figure shows the adhesion of the PVC complex nutrients to the intestinal wall). This was not observed in the group of rats receiving the common vitamins.

To further investigate the absorption of vitamins, we investigated the pharmacokinetics and biodistribution over 8 h. Tail blood was drawn every 2 hours after oral administration to healthy rats and the fluorescence concentration in plasma samples was determined. As shown in fig. 6, the time scale of the peak concentration of each PVC group in plasma was significantly longer than that of the corresponding unmodified vitamin group, while the area under the plasma concentration-time curve exceeding 8h was not reduced, which fully indicates that the absorption rate of the PVC-modified vitamin in the intestinal tract was significantly improved.

In order to explore the absorption effect of minerals in the PVC vitamins, oil-soluble vitamins and PVC water-soluble vitamins are mixed into PVC _ADE Solution (2mL, 2mgmL) ^-1 ) And orally administered to healthy rats, and the rats were gavaged with normal vitamin mineral nutrients as a control group. Blood is drawn for 3h respectively for blood vitamin content detection. As shown in FIG. 7, by observing the blood concentration of rats fed the vitamin mineral complex supplement for 3 hours, it was found that the content of Fe in the plasma of the PVC group was significantly higher than that of the ordinary vitamin mineral complex group, which fully illustrates that the absorption rate in the intestinal tract is significantly improved by the PVC-modified vitamins and minerals.

Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (6)

1. A method for enhancing the absorption of vitamins and minerals in intestinal tracts is characterized in that plant polyphenol and minerals are complexed to form a nano-network material, the nano-network material is combined with vitamin molecules, and a polyphenol-based vitamin compound is formed through the wrapping or agglomeration adsorption; the polyphenol-based vitamin complex has long retention time in gastrointestinal tract, and can promote absorption of vitamins and minerals in gastrointestinal tract.

2. The method of enhancing the absorption of vitamins and minerals in the intestinal tract of claim 1 wherein said mineral is zinc ion Zn ²⁺ Or ferrous ion Fe ²⁺ 。

3. The method of claim 1, wherein the plant polyphenol is one or more of tannic acid, epigallocatechin gallate, rutin, and anthocyanins.

4. The method of enhancing the absorption of vitamins and minerals in the intestinal tract according to claim 1 wherein said vitamin is selected from the group consisting of oil soluble vitamin a, vitamin D, vitamin E, and mixtures of water soluble vitamin C, vitamin B2, vitamin B12, calcium pantothenate, folic acid, and mixtures of one or more thereof.

5. The method of enhancing the absorption of vitamins and minerals in the intestinal tract according to claim 4, wherein said polyphenol based vitamin complex is prepared for oil soluble vitamins by:

(1) Dissolving oil-soluble vitamins in soybean oil to prepare an oil agent, and adding the oil agent into a phosphate buffer solution for ultrasonic treatment to form a vitamin emulsion;

(2) Dissolving plant polyphenol in ultrapure water to form a polyphenol water solution; and will contain Zn ²⁺ Or Fe ²⁺ Dissolving the soluble salt in another part of ultrapure water to obtain a mineral water solution;

(3) Adding vitamin emulsion into polyphenol water solution, performing ultrasonic treatment, adding mineral water solution, mixing, centrifuging, and collecting upper emulsion to obtain polyphenol-based vitamin complex.

6. The method of claim 4, wherein the polyphenol based vitamin complex is prepared for a water soluble vitamin by:

(1) Adding water-soluble vitamins into a phosphate buffer solution to form a vitamin solution;

(2) Dissolving plant polyphenol in ultrapure water to form a polyphenol water solution; and will contain Zn ²⁺ Or Fe ²⁺ Dissolving the soluble salt in another part of ultrapure water to obtain a mineral water solution;

(3) Adding a vitamin solution into a polyphenol water solution, and stirring at room temperature to ensure that the vitamin is fully contacted with the plant polyphenol to form nanoparticles by agglomeration; then adding mineral water solution, stirring well, dialyzing the obtained solution under low concentration phosphate buffer solution for 2 days to remove impurities, and cutting off 3.0kDa of molecular weight; and freeze-drying the obtained polyphenol-based vitamin compound solution to prepare polyphenol-based vitamin compound dry powder.

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