JP2747935B2 - Manufacturing method of iron-fortified beverage - Google Patents

Description

Description: BACKGROUND OF THE INVENTION The present invention relates to an iron-enriched lactoferrin that has various physiological activities and maintains the physiological functionality of lactoferrin, while increasing the absorption of iron into the body by allowing iron to coexist with lactoferrin. The present invention relates to a method for producing a beverage.

Thus, this beverage is useful for iron supplementation and physiological functions.

Prior art According to the National Nutrition Survey of 1987, the average intake of iron by Japanese people is 98% of the average required amount, which is somewhat insufficient.

In particular, it is said that there are a large number of anemia and anemia reserve groups (potential iron deficiency) due to insufficient iron intake and increased iron excretion among young women and athletes.

By the way, those with a potential iron deficiency do not have any particular problems in their normal behavior, but are susceptible to anemia symptoms when they exercise vigorously or when they are stressed strongly.

As described above, despite the lack of iron intake, there is a problem, however, unless the symptoms of anemia appear, they will not receive the iron preparation as a pharmaceutical, and even if they are somewhat conscious of anemia, they will take conventional iron supplements Doing so is reluctant because of their palatability, and is therefore less likely to consume iron supplements.

By the way, there are several iron supplements currently available on the market in the form of beverages, but most of them use inorganic iron salts (such as sodium ferrous citrate) as iron agents, and in the case of inorganic iron, The absorption rate in the body is said to be about 5%, so it is hard to say that it is an effective iron supplement. In addition, in forms other than beverages, some foods contain heme iron, which is said to have a high absorption rate.However, due to the problem of flavor, processing such as encapsulation is performed, and as a result, costs are reduced. It is expensive, and it is hard to say that it is food.

Recently, a hematopoietic agent containing an iron-binding lactoferrin obtained by chelating a ferric ion to lactoferrin isolated from milk has been developed, and this hematopoietic agent can be applied as a food material for various foods and drinks. It has been proposed (JP-A-63-22525).

However, when the above-mentioned hematopoietic agent is added to a beverage and used by dissolving, the lactoferrin precipitates by heating, in which the three-dimensional tertiary structure is destroyed by heating,
There is a problem that its original physiological function is impaired, and the ability to chelate iron is also lost.

The standard for soft drink production under the Food Sanitation Law is pH 4.0.
If it is less than 65 ° C, it must be heated for 10 minutes or sterilization with the same or more effect, and if it is pH 4.0 or more, it must be heated at 85 ° C for 30 minutes or sterilization with the same or more effect. It is specified.

Problems to be Solved by the Invention The present invention, by using an iron agent in the coexistence of lactoferrin, good absorption of iron into the body, and the aqueous solution in which the iron agent and lactoferrin coexist, the ionic strength of the specific An object of the present invention is to provide a method for producing an iron-enriched beverage having a physiological function of lactoferrin and having no problem in flavor by sterilizing by heating under conditions.

Means for Solving the Problems Lactoferrin used in combination with an iron agent in the present invention is an iron-binding glycoprotein present in exocrine fluid such as milk, and is a nutritionally and pharmacologically important milk protein. .

Conventionally, as a method of separating and purifying lactoferrin, a method using an ion exchange resin (Gorden et al., Biochim. Biop.
hys. Acta, 60: 410-411, 1962), a method using heparin affinity chromatography (Blockberg et al., FEB)
S Lett., 109: 180, 1980), a method using an anti-lactoferrin monoclonal antibody-immobilized column (JP-A-61-145200).
Japanese Patent Application Laid-Open No. 63-255300) and a method using a sulfated carrier (Japanese Patent Application Laid-Open No. 63-255300) are known. Lactoferrin obtained by any method can be used.

In the present invention, a mixed solution of lactoferrin and an iron agent and sodium bicarbonate or an aqueous solution of iron lactoferrin is prepared, and the ionic strength (I) of the solution is adjusted under the conditions shown in the following formula, followed by heat sterilization. Note that the sodium bicarbonate used here is used to increase the affinity between lactoferrin and iron, but a small amount may be used, so that a trace amount of sodium bicarbonate contained in tap water may be used.

log I ≦ −1.3 (T ≧ 80) (T represents heat sterilization temperature (° C.)) In the present invention, a solution to which raw materials for beverages such as acidulants, flavors, sugars, vitamins, etc. are added is lactoferrin, iron agent and bicarbonate. Even when mixed with an aqueous solution in which sodium is dissolved or an aqueous solution in which lactoferrin iron is dissolved, if it can be adjusted under the conditions of the ionic strength I, lactoferrin and a raw material for beverages are mixed, and this is mixed under the conditions of ionic strength I. It may be adjusted, heat-sterilized, cooled, filled into a sterilized container, and supplied as an iron-enriched beverage.

Further, when the solution to which the beverage raw material is added does not satisfy the condition of the ionic strength I, or when the solution to which the beverage raw material is added cannot be adjusted to the condition of the ionic strength I, lactoferrin, iron agent and sodium bicarbonate are used. An aqueous solution in which lactoferrin iron is dissolved or an aqueous solution in which lactoferrin iron is dissolved is adjusted under the condition of the ionic strength I, and the solution in which the beverage raw material is dissolved is separately heat-sterilized. After cooling, the two solutions are mixed and sterilized. What is necessary is just to fill in the filled container and to supply it as an iron-fortified beverage.

In this way, by adjusting the ionic strength of the lactoferrin-containing solution and sterilizing by heating, lactoferrin can be heated in a state of low ionic strength, so that lactoferrin can be prevented from denaturing and its functionality can be maintained. Becomes

FIG. 1 shows the relationship between the ionic strength and heating temperature of the aqueous lactoferrin solution and the iron binding ability.

These are those in which the ionic strength of the aqueous solution of lactoferrin was adjusted, kept at each heating temperature for 30 minutes, and immediately cooled, and the iron binding ability after cooling was indicated by% with respect to the heated product.

When T is lower than 60 ° C., lactoferrin is not denatured. Therefore, it is not necessary to adjust the ionic strength I.
When the temperature is higher than 60 ° C., the ionic strength I
If it is removed, precipitation of lactoferrin occurs or iron binding ability decreases. Therefore, when heat sterilizing an aqueous lactoferrin solution having a high salt concentration, it is necessary to adjust the ionic strength of the aqueous solution by desalting so that the ionic strength I meets the conditions shown in the above equation.

In addition, in order to heat-sterilize a lactoferrin aqueous solution having a low salt concentration in a state in which other beverage ingredients are mixed, it is necessary to adjust the ionic strength I so as to meet the above formula and then heat-sterilize.

In the present invention, the content of lactoferrin in the beverage is suitably 0.1 to 6.0% (that is, a concentration of 0.1 to 6 g / 100 ml). The content of lactoferrin in the beverage is based on the following required amount of coexisting iron. That is,
Since the amount of iron provided by lactoferrin due to its iron-binding function is 1.5 mg per 1 g of lactoferrin, if the content of lactoferrin in the beverage is less than 0.1%, the amount of iron provided as beverage is 100%.
It is not practical because it is necessary to consume 700 ml or more as a beverage in order to replenish iron which is 0.15 mg or less per ml, which is equivalent to the generally known iron excretion of 1.0 mg per day.

On the other hand, when lactoferrin is contained in the beverage in an amount exceeding 6%, the viscosity of a mixed solution of lactoferrin, an iron agent and sodium bicarbonate increases, so that diafiltration (diafiltration) for adjusting the ionic strength (I) of the solution is performed.
tration) slows down the transmission speed, which is not advisable for adjusting the ion velocity (I).

In addition, dissolve iron lactoferrin and other beverage ingredients,
Even in the method of heat sterilization after mixing, when the lactoferrin concentration becomes 6% or more, the liquid viscosity increases, the amount of lactoferrin increases, and it takes a long time to dissolve. The lactoferrin concentration is preferably 6% or less.

In addition, when the content of iron lactoferrin is 6%, the iron content in the beverage is 9 mg / 100 g, and the iron absorption rate of iron lactoferrin is sufficient for iron supplementation even when iron lactoferrin is 6% or less due to the high iron absorption rate of iron lactoferrin.

The iron agent or lactoferrin iron used in the present invention is preferably contained as iron in an amount of 0.15 to 12.0 mg per 100 ml of beverage. If the content of the iron agent is less than 0.15 mg,
As mentioned above, it is necessary to collect 700 ml or more of beverages in order to secure an amount of iron equivalent to 1.0 mg of iron excretion per day. Is not preferred.

Examples Hereinafter, the present invention and its effects will be specifically described with reference to examples.

Example 1 Preparation of lactoferrin: A commercially available crosslinked chitosan (trade name), sulfated chitopearl 10 obtained by sulfate esterification with sulfuric anhydride by a conventional method, was packed in a column having a diameter of 32 cm and a length of 200 cm. The defatted milk 500 was passed through the column for 200 hours. Next, the column was washed with 50 of 0.3 M saline, and then lactoferrin adsorbed on the column was eluted with 30 of 1.0 M saline. The resulting lactoferrin solution is
After desalting using an ED device (TS-210, manufactured by Tokuyama Soda Co., Ltd.), the solution was concentrated 10-fold with a UF device (DB-2, manufactured by Amicon) and freeze-dried.

The recovered lactoferrin was 6 g, and the purity was determined to be 95%. The iron binding amount was measured to be 0.2 mg Fe / g protein, and it was confirmed that the iron binding ability was 98%.

Lactoferrin and iron-containing solution (A)
And a raw material solution (B) for a beverage.

Solution A: (g) lactoferrin 600 sodium bicarbonate 86 ferric chloride hexahydrate 10 is dissolved in water to make 50.

Solution B: (g) Sodium iron citrate 48 Isomerized sugar 8000 Citric acid 800 Sodium citrate 800 Vitamin B ₂ 1.1 Vitamin B ₆ 2.2 Vitamin C 5.5 Folic acid 0.4 Fragrance 100 1/5 Dissolve 1000 concentrated juice 1000 in water I do.

The solution A was subjected to diafiltration to remove low molecular ions, and the ionic strength was adjusted to 10 ^-3 or less. Then, the solution A was sterilized by heating (holding at 93 ° C. for 3 minutes), and then cooled (5 to 10). C) was stored in a sterilized tank.

On the other hand, the solution B was sterilized by heating in the same plate (to 93 ° C).
The mixture was kept for 20 seconds) and then cooled (5 to 10 ° C.) and added to the above-mentioned stock solution A and mixed.

Next, this mixed solution was filled in a sterilized paper container in a sterile room to obtain an iron-enriched beverage.

The resulting beverage (pH 3.8, iron content 5.81 mg / 100 ml) was also good in flavor.

Example 2 Using lactoferrin prepared in the same manner as in Example 1,
Lactoferrin 600g, sodium bicarbonate 86g, second chloride
10 g of iron hexahydrate was dissolved in water to make 50, and a solution containing lactoferrin and iron was prepared. Next, this aqueous solution is subjected to diafiltration treatment to remove low molecular ions,
After adjusting the ionic strength to 0.001 or less, further add 16 g of sodium iron citrate, 10,000 g of sucrose, 100 g of flavor, and 7000 g of 1/5 frozen concentrated juice (Bx45), and add water to make 100 to prepare an iron-fortified beverage. did.

The thus prepared iron-enriched beverage is heat-sterilized (93 ° C, 2-3 seconds) with a plate-type sterilizer, then bottled, inverted, held for 20 seconds, and then cooled (5-10 ° C). The product of iron fortified beverage was obtained.

The resulting iron-enriched beverage had a pH of 3.7, an ionic strength of 0.02, and an iron content of 2.4 mg / 100 ml.

Example 3 Sulfated chitopearl was regenerated by passing 2.0 M saline and 0.15 M saline through the sulfated chitopearl used in Example 1.

This regenerated sulfated chitopearl 10 and cheese whey 1000
Was mixed with stirring for 1 hour, and then the gel-like mixture was added to 0.1 g.
After washing with 3M saline 50, lactoferrin adsorbed on the gel mixture was eluted with 1.0M saline 30. After adding 500 mg of ferric chloride to the obtained lactoferrin solution, excess ions were removed by passing through an ion exchange resin used for desalting, and spray-dried to obtain iron-saturated bovine lactoferrin 48.
g was obtained. The recovered bovine lactoferrin was 95% pure and the amount of iron bound was determined to be 1.3 mg Fe / g protein. It was confirmed that the iron saturation was 93% or more.

Using the lactoferrin iron prepared as described above, an iron-enriched beverage was prepared according to the following formulation.

Formulation: (g) Iron lactoferrin 600 Sucrose 11000 Fragrance 100 Sodium iron citrate 16 Frozen concentrated juice (Bx45) 12000 was dissolved in water to make 100.

Heat this iron-fortified beverage in a jacketed tank,
After heat sterilization at 15 ° C. for 15 minutes, the product was filled in a sterilized bottle to obtain an iron-reinforced product.

The obtained iron-fortified beverage had a good taste and a good color at pH 3.3, ionic strength of 0.058, and iron content of 2.5 mg / 100 ml.

Example 4 Using lactoferrin iron prepared in the same manner as in Example 3, an aqueous lactoferrin solution (A) and a raw material solution for beverages (B) were prepared according to the following formulation.

Solution (A) Dissolve 600 g of iron lactoferrin to make 50.

Solution (B) Same composition as in Example 1.

Hereinafter, an iron-enriched beverage was obtained according to the same procedure as described in Example 1.

Example 5 This method shows the results obtained by measuring the transition of lactoferrin activity during the production process of Example 1 by an immunological method utilizing a complement fixation reaction. The results are as shown in Table 2.

As can be seen from Table 2, the activity of lactoferrin in the product beverage was reduced by 13% as compared to that before the formulation, but such a decrease did not substantially affect the function of lactoferrin.

Example 6 This example shows the effect of the iron-fortified beverage according to the present invention on the recovery from anemia.

A 6-week-old female Wistar rat fed an iron-free feed (0.25 mg of iron / 100 g of feed) for 3 weeks and brought into anemia (having a hemoglobin [Hb] concentration of 8 g / dl or less) obtained by the drink obtained in Example 1 (Iron content 5.81 mg / 100 ml), except for lactoferrin, except that sodium iron citrate was used as an iron agent, and a beverage administration group prepared according to the procedure described in Example 1 and a commercial feed (iron 20 mg / 100 g). ), And further into each administration group of iron-free diet and deionized water only. Then, except for the group administered with the commercial feed, the group was divided into three groups so that the average blood [Hb] concentration was similar, and the group administered with the beverage of the present invention and a lactoferrin-free beverage using an iron agent of sodium iron citrate were used. Was administered orally every day at a concentration of 50 μg / day / animal as iron.

Blood is collected every two weeks after the start of the administration of iron, and the [Hb] concentration,
The hematocrit [Ht] value and the number of red blood cells [RBC] were measured.

(Hb) concentration, hematocrit (Ht) value, erythrocyte (RB
C] The measurement results of the numbers are shown in FIGS. 2, 3 and 4.

As can be seen from each figure, the administration group of the iron-enriched beverage according to the present invention obtained in Example 1 was equivalent to the administration group of the iron-containing commercial feed in all the measured values, and the recovery ability of anemia was improved. It was found to be high. On the other hand, it can be seen that the effect of anemia recovery is extremely low even in 70 days after the administration of the beverage using only sodium iron citrate containing no lactoferrin as an iron agent.

Effect of the Invention According to the present invention, it is possible to obtain an iron-enriched beverage that has high absorbability of iron into the body and maintains the physiological function of lactoferrin coexisting with iron.

[Brief description of the drawings]

FIG. 1 shows the relationship between the ionic strength and the heating temperature of the aqueous lactoferrin solution and the iron binding ability. FIG. 2 shows the daily change of the blood [Hb] concentration when the drink according to the present invention and each comparative example were administered, FIG. 3 similarly shows the daily change of the [Ht] value, and FIG. RBC] number over time.

 ──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-2-108629 (JP, A) JP-A-63-22525 (JP, A)

Claims (4)

(57) [Claims] When an iron-enriched beverage containing iron-bound lactoferrin is produced, an aqueous solution in which lactoferrin, an iron agent and sodium bicarbonate are dissolved, or an aqueous solution in which lactoferrin iron is dissolved is used. A method of producing an iron-enriched beverage, which comprises heat sterilizing and cooling the mixture, heat sterilizing a solution of the beverage raw material, cooling, and then mixing the two solutions. log I ≦ −1.3 (T ≧ 80) (T indicates heat sterilization temperature ° C) 2. When an iron-enriched beverage containing iron-bound lactoferrin is produced, lactoferrin, an iron agent and sodium bicarbonate are dissolved, and an aqueous solution in which other beverage ingredients are dissolved or iron lactoferrin is dissolved. A method for producing an iron-enriched beverage, comprising adjusting an ionic strength I of an aqueous solution in which other beverage raw materials are dissolved to the condition shown in the following formula, and then sterilizing by heating. log I ≦ −1.3 (T ≧ 80) (T indicates heat sterilization temperature ° C) 3. The beverage according to claim 1, wherein lactoferrin is used in an amount such that the beverage has a concentration of 0.1 to 6 g / 100 ml.
3. The method for producing an iron-enriched beverage according to item 1. 4. A beverage containing iron as iron in an amount of 0.15-12.0 mg / 100 ml in a beverage.
(1) to (3), which are used in such an amount as to give a concentration of
The method for producing an iron-fortified beverage according to any one of the above.