JP2743178B2 - How to use adsorbent for lipid - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a selective lipid adsorbent.

The modernization of living conditions and the advancement of food production technologies have allowed human beings to extend their life expectancy in recent decades. This trend has been pioneered by developed nations in Europe and the United States, but it will be one of the most important issues for humanity as a whole. With the extension of life expectancy, adult illness must be focused on as a new social problem. Among adult diseases, various approaches have been tried for cardiovascular diseases from the viewpoint of preventive medicine and from the viewpoint of clinical diagnosis. However, it is good to say that there is no definitive treatment at the clinical site so far.

Based on the above, the present inventors have conducted intensive studies and found that selective lipid adsorption that adsorbs lipids such as cholesterol and β-lipoprotein, which can adversely affect living organisms from foods as well as body fluids such as blood. Came to discover the drug.
Hereinafter, the HDL cholesterol, α-lipoprotein, free fatty acid, and the lipid adsorbent that does not adsorb albumin will be described in detail.

The present invention relates to sterols and triglycerides (neutral fats) such as cholesterol present in body fluids and foods in living bodies,
Among various lipids such as phospholipids, cholesterol esters, fatty acids, fatty acid esters, glycolipids, etc., which is useful for living organisms, and which is a lipid adsorbent having selectivity not adsorbing HDL cholesterol, α-lipoprotein and free fatty acids. It is. Further, since this adsorbent does not adsorb protein, albumin, it can be applied not only to medical fields including medical aids and blood products, but also to food fields such as food ingredient improvement processing.

The components constituting the present agent are composed of a calcium phosphate compound and a phosphorus-containing compound. Specific examples of the calcium phosphate compound include hydroxyapatite, tricalcium phosphate, tetracalcium phosphate, calcium pyrophosphate and the like. Bone charcoal containing a calcium phosphate compound can also be used. Phosphorus-containing compounds include, for example, phosphates such as sodium phosphate and potassium phosphate, phosphoric acid, phosphorus-containing organic substances such as phosphate ions and phosphocitidylcholine, phosphorus-containing inorganic substances such as pyrophosphate, phosphate buffers and the like. Are specifically exemplified.

HDL cholesterol, α-
In order to have a selective lipid adsorption function that does not adsorb lipoproteins, free fatty acids or albumin, phosphate ions must be present in the environment where the calcium phosphate compound and the adsorbed substance are present. Inorganic acids such as hydrochloric acid and sulfuric acid having an action of releasing acetic acid and organic acids such as citric acid, succinic acid and acetic acid can also be constituents of the lipid adsorbent. Further, a substance obtained by treating a calcium phosphate compound and / or a material containing the calcium phosphate compound with the above-described inorganic acid and organic acid can also be used as a component of the lipid adsorbent.

Examples of the calcium phosphate compound include hydroxyapatite, tricalcium phosphate, tetracalcium phosphate, calcium pyrophosphate, and the like. Hydroxyapatite had excellent selective adsorption of lipids. Calcium phosphate having a molar ratio of calcium to phosphorus of 1 to 2 can be preferably used, and calcium phosphate having a molar ratio of calcium to phosphorus of 1.54 to 1.68 can be more preferably used.

The lipid adsorbent of the present invention is a major cause of cardiovascular diseases without selectively adsorbing proteins such as albumin and HDL cholesterol, α-lipoprotein and free fatty acids which are useful for living organisms. Since it has excellent selectivity for adsorbing lipids such as cholesterol at a high rate, the following use examples can be presented.

(1) Component improvement treatment of blood products.

(2) A blood purification method that selectively adsorbs and removes lipids such as cholesterol from body fluids such as blood and returns them to the living body without adsorbing proteins such as albumin and useful lipids such as HDL cholesterol, α-lipoprotein and free fatty acids. Filtering agents used for medical devices such as medical devices or medical aids.

(3) Used as a column for lipid analysis and fractionation for chromatograms such as HPLC used to analyze lipids. Further, it is used as a precolumn for removing lipid components when analyzing body fluid components.

(4) It is used as a treatment for improving foods such as oils and fats, soy milk, cheese, yogurt, milk, dairy products, chicken eggs, and food ingredients.

(5) Since the components constituting the present invention are extremely safe for the human body, they are used as functional foods as an element of food components.

(6) Pharmaceuticals.

(7) Filter media used for recycling edible oil.

(8) Filtering agents used for oil drainage processing and crude oil refining.

 Next, the present invention will be described in detail with reference to examples.

Experimental Example 1 Synthetic hydroxyapatite (HA) crushed by Ca / P 1.657, attrition mill and calcined at 800 ° C. was used, and adsorption was performed by a batch method.

1) (Untreated group) 200 mg of HA was reacted with 1 ml of serum or plasma supplemented with sodium heparin, and the adsorption rate was determined from the decrease in lipid before and after adsorption.

2) (Treatment group) 100 μL of 0.5 M phosphate buffer (PB) of pH 7.4 was added to 1 ml of serum and plasma, and the adsorption rate was similarly determined. Quantification of blood lipids was performed using a Hitachi automatic analyzer. Lipid fractionation was performed by electrophoresis.

The results are shown in Table 1. Table 1 (a) shows the adsorption rate of the untreated group, and Table 1 (b) shows the treated group.

From this, 1) In the PB untreated group, the adsorption rate of serum components to HA
6.3%, albumin is 7.8%, lipid adsorption is total cholesterol 12%, free cholesterol 8.9%, ester cholesterol 13%, triglyceride 0.37%, β-lipoprotein 6.6%, HDL cholesterol 30%, phospholipid 18
%, Free fatty acids -4.2%.

2) Treatment group with phosphate buffer added total protein 1.3
%, Albumin -2.1% and protein, especially albumin adsorption decreased, lipid adsorption, total cholesterol 82%,
Free cholesterol 85%, ester cholesterol 82
%, Triglyceride 83%, β-lipoprotein 98%, HDL cholesterol -5.6%, phospholipid 59%, free fatty acid 1.5%
In addition, adsorption of lipids other than HDL cholesterol and free fatty acids increased remarkably. Almost the same results were obtained with plasma.

Experimental Example 2 In order to investigate that the specificity of HDL cholesterol and total cholesterol adsorption to hydroxyapatite (HA) changes depending on the phosphate ion concentration, the experiment was carried out by changing the phosphate buffer concentration in the reaction solution. Was.

HA was synthesized by a wet method, dehydrated and dried, then pulverized by a sample mill, and further baked at 800 ° C.

Using human serum as a sample, 0.1 ml of various concentrations of phosphate buffer (pH 6.4) was added to 0.9 ml of serum to make the total volume 1 ml.

The phosphate buffer concentration after the reaction is 0,5,10,2 as the final concentration.
The concentration was adjusted to 0,50,100 mM, 200 mg of the above HA was added thereto, and the mixture was reacted at room temperature for 10 minutes. Then, the total cholesterol and HDL cholesterol in the centrifuged supernatant were measured. The results are shown in Table 2.

Example 3 0.1 ml of distilled water or 0.1 ml of 0.5 M phosphate buffer (pH 7.4) was added to 0.9 ml of human serum, and 200 mg of the HA shown in Experimental Example 2 was added to each sample having a total volume of 1 ml. After gently stirring at room temperature for 10 minutes, the centrifuged supernatant was measured by Corning and a lipoprotein analysis system manufactured by Helena.

The results are shown in FIG. Figure 1a shows 0.1 ml of 0.5M in 0.9 ml of the original serum.
The distribution of lipoprotein when a phosphate buffer (pH 7.4) is added and HA is not added is shown. Figure 1b shows HA in the absence of phosphate buffer.
Fig. 1c shows the distribution of non-adsorbed lipoprotein in the presence of 50 mM phosphate buffer.

Experimental Example 4 The molar ratio of calcium to phosphorus in calcium phosphate (Ca /
In order to compare the specific adsorption specificity of lipids due to the difference in P), the adsorption of serum HDL cholesterol and total cholesterol to calcium phosphate was examined.

200 mg / ml of serum in the presence of 50 mM phosphate buffer (pH 7.4)
Was added with calcium phosphate having a Ca / P of 1.54, 1.56, 1.66, and 1.68, and the mixture was slowly stirred at room temperature for 10 minutes. The results are shown in Table 3.

As described above, as is clear from the experimental examples, the lipid adsorbent of the present invention is not limited to lipids contained in foods as well as body fluids such as blood, and may contain triglycerides, free cholesterol, β- Adsorbs almost all lipids such as lipoproteins, but has been shown to be useful for living organisms.
-The result was that there was excellent adsorption specificity that does not adsorb lipids such as lipoproteins and free fatty acids and proteins such as albumin.

Also, since the agent of the present invention is completely harmless to living organisms,
It can be effectively used not only as a pharmaceutical product but also as a method for improving food and food ingredients.

[Brief description of the drawings]

FIG. 1 shows the results of examining the adsorption specificity of serum lipoprotein to HA, wherein a shows the electrophoresis pattern of the original serum control, and b and c show the electrophoretic patterns of the HA non-adsorbed fraction. The horizontal axis shows the migration distance from the origin, and the vertical axis shows the absorbance. Α shown in the figure
-Lipo indicates α-lipoprotein, and β-Lipo indicates β-lipoprotein.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI G01N 30/88 G01N 30/88 C (72) Inventor Shunpei Inventor 5-7-33-101 Heights Ito, Nakameguro, Meguro-ku, Tokyo ( 72) Inventor Shinichiro Kusunoki 2-23-21 Oizumi Gakuen-cho, Nerima-ku, Tokyo Examiner Eiko Ebihara (56) References JP-A-63-162794 (JP, A) JP-A-63-97696 (JP, A)

Claims (1)

(57) [Claims] 1. A method of using an adsorbent for lipids, wherein a calcium phosphate compound is used in the presence of a phosphorus-containing compound as an adsorbent having no adsorptivity for HDL cholesterol, α-lipoprotein and free fatty acids.