CN109293802B - Agar oligosacchride iron and preparation method thereof - Google Patents
Agar oligosacchride iron and preparation method thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0036—Galactans; Derivatives thereof
- C08B37/0039—Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Abstract
The invention discloses agar oligosacchride iron and a preparation method thereof, which takes gracilaria agar as a raw material and prepares the agar oligosacchride iron by refining, acid degradation, high-voltage pulse electric field assisted chelation, membrane separation, freeze drying and crushing. Refining to obviously improve the content of neutral sugar, and promoting the chelation of agar oligosaccharide and iron ions under the catalysis of a high-voltage pulse electric field after acid degradation. The invention has advanced and reasonable technology and high production efficiency; the prepared agar oligosaccharide iron product has high iron content, and has good oxidation resistance, so that the cell activity can be improved, the absorption of the organism to iron can be promoted, and the bioavailability is high.
Description
Technical Field
The invention relates to the field of food processing, in particular to a preparation method of agar oligosaccharide iron.
Background
Iron is one of the essential trace elements of human body, and it is an essential component constituting heme, and plays an important role in oxygen transport, deoxyribonucleic acid synthesis, and energy metabolism in human body. Iron deficiency can lead to iron deficiency anemia or dysfunction. Studies have shown that iron deficiency exists in 30% of the population worldwide, 46% of children aged 5-14 years and 48% of pregnant women suffer from iron deficiency anemia. Currently, the prevention and treatment of iron deficiency anemia is mainly realized by taking an iron supplement. Common iron supplement agents comprise ferrous sulfate, ferrous chloride, ferrous gluconate and the like, and although the iron supplement agents have high iron content, the iron supplement agents have poor stability, are easy to generate free iron and generate free radicals in vivo, so that lipid peroxidation is caused; and has special metal rust taste, bad taste and is easy to cause gastrointestinal adverse reactions.
The agar oligosaccharide has good water solubility and is easy to be absorbed by human body; the agar oligosaccharide contains a large amount of hydroxyl and carboxyl, and can form a stable coordination effect with metal ions, so that the agar oligosaccharide is an ideal material for processing iron chelates, and on one hand, the stability of products can be improved, and on the other hand, the agar oligosaccharide has good oxidation resistance and can activate cells, thereby promoting the absorption of organisms on iron.
Therefore, the agar oligosaccharide iron chelate and the preparation method thereof developed by the invention have important significance for improving the iron absorption and utilization rate of organisms and have good application prospects.
Disclosure of Invention
The invention aims to provide agar oligosaccharide iron and a preparation method thereof aiming at the outstanding problems of low bioavailability and single function of the existing iron supplement product, wherein the product has high iron content and high bioavailability.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing agar oligosaccharide iron comprises the following steps: refining gracilaria agar, performing acid degradation, performing high-voltage pulse electric field assisted chelation, performing membrane separation, freeze drying and crushing to obtain the agar oligosaccharide iron.
The method comprises the following specific steps:
(1) refining gracilaria agar: preparing gracilaria agar into 3-5 wt% glue solution by using ultrapure water, heating to 90 ℃, stirring until the glue solution is completely dissolved, adding 10-20% of polyethylene glycol 6000 and 1% of NaCl clarifying agent by mass ratio to the glue solution into the glue solution, treating for 3-8 min at 20-40 kHz by using an ultrasonic vibrator, cooling to room temperature, standing for precipitation, centrifuging, taking separation liquid, performing the same clarification and ultrasonic vibration treatment, standing for precipitation, centrifuging, combining precipitates, washing the precipitates twice by using cold water, and performing filter pressing and dehydration to obtain refined agar;
(2) acid degradation: preparing refined agar into a glue solution with the concentration of 3-5 wt% by using ultrapure water, heating to 90 ℃, stirring until the refined agar is completely dissolved, adding an HCl solution with the final concentration of 1.0mol/L, treating for 90min, dropwise adding a 1.0mol/L NaOH solution until the pH value is 7, centrifuging at 7000 r/min for 15min, and taking a supernatant to obtain an agar oligosaccharide hydrolysate;
(3) high-voltage pulse electric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2O and trisodium citrate dihydrate, wherein the mass ratio of agar oligosaccharide (in dry weight) to iron ions to trisodium citrate dihydrate is 7.5-2.5: 1: 7.5-0.83, the pH is adjusted to 3-7 by adopting HCl or NaOH, the mixture is heated to 50-80 ℃, and then treated for 30-60 min by adopting a high-voltage electric field of 20-60 kV/cm, so as to obtain a reaction solution;
(4) membrane separation: filtering the reaction solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3000Da, collecting the permeate, and filtering the permeate by using an ultrafiltration membrane with the molecular weight cutoff of 500Da to obtain the retentate, namely the agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating the agar oligosaccharide iron solution, placing the concentrated agar oligosaccharide iron solution in a vacuum freeze dryer, drying the agar oligosaccharide iron solution until the water content is below 5wt% at the absolute pressure of 20Pa in a drying chamber, the pre-freezing temperature of-35 ℃ and the resolution temperature of 35 ℃, and sieving the crushed agar oligosaccharide iron solution with a 80-mesh sieve to obtain the agar oligosaccharide iron.
The invention has the following remarkable advantages:
(1) polyethylene glycol 6000 and NaCl are adopted, ultrasonic vibration treatment is combined, impurities and non-neutral sugar in the gracilaria agar are effectively removed, and a material foundation is laid for preparing a high-quality agar oligosaccharide iron product; (2) the high-voltage pulse electric field is adopted to catalyze the chelation of the agar oligosaccharide and iron ions, so that the chelation time is effectively shortened, and the chelation rate is improved; (3) the separation of high-activity agar oligosaccharide iron is realized by adopting secondary membrane separation, and the bioavailability of the product is ensured; (4) the agar oligosaccharide iron chelate prepared by the invention has good stability, and the agar oligosaccharide has good oxidation resistance and can activate cells, thereby promoting the absorption of organisms to iron.
Drawings
Fig. 1 is an infrared spectrum of agar oligosaccharide iron and agar oligosaccharide.
Agar oligosaccharide at 1725 cm-1The absorption peak of carboxyl is shown, and the absorption peak disappears in the spectrum of the agar oligosaccharide iron, which shows that the carboxyl in the agar oligosaccharide can participate in the chelation reaction as the chelation site with the iron in the ferric trichloride. In addition, compared with agar oligosaccharide, the iron content of agar oligosaccharide is 3385 cm-1Has a sharper absorption peak at 1605 cm-1And 1386 cm-1The absorption peak of (A) was shifted to 1636 cm respectively-1And 1412 cm-1And the hydroxyl is involved in the chelation of the agar oligosaccharide and the iron. Particularly, 853, 640, 594 and 556 cm appear in the iron spectrum of agar oligosaccharide-1Four new peaks in the fingerprint area (1000-400 cm) of carbohydrate-1) Are characteristic of beta-FeOOH. Thus indicating that the agar oligosaccharide and the iron have chelation reaction.
Detailed Description
The method is realized by the following technical steps:
a method for preparing agar oligosaccharide iron comprises the following specific steps:
(1) refining gracilaria agar: preparing gracilaria agar into 3-5 wt% glue solution by using ultrapure water, heating to 90 ℃, stirring until the glue solution is completely dissolved, adding 10-20% of polyethylene glycol 6000 and 1% of NaCl clarifying agent by mass ratio to the glue solution into the glue solution, treating for 3-8 min at 20-40 kHz by using an ultrasonic vibrator, cooling to room temperature, standing for precipitation, centrifuging, taking separation liquid, performing the same clarification and ultrasonic vibration treatment, standing for precipitation, centrifuging, combining precipitates, washing the precipitates twice by using cold water, and performing filter pressing and dehydration to obtain refined agar;
(2) acid degradation: preparing refined agar into a glue solution with the concentration of 3-5 wt% by using ultrapure water, heating to 90 ℃, stirring until the refined agar is completely dissolved, adding an HCl solution with the final concentration of 1.0mol/L, treating for 90min, dropwise adding a 1.0mol/L NaOH solution until the pH value is 7, centrifuging at 7000 r/min for 15min, and taking a supernatant to obtain an agar oligosaccharide hydrolysate;
(3) high voltage pulseElectric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2O and trisodium citrate dihydrate, wherein the mass ratio of agar oligosaccharide (in dry weight) to iron ions to trisodium citrate dihydrate is 7.5-2.5: 1: 7.5-0.83, the pH is adjusted to 3-7 by adopting HCl or NaOH, the mixture is heated to 50-80 ℃, and then treated for 30-60 min by adopting a high-voltage electric field of 20-60 kV/cm, so as to obtain a reaction solution;
(4) membrane separation: filtering the reaction solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3000Da, collecting the permeate, and filtering the permeate by using an ultrafiltration membrane with the molecular weight cutoff of 500Da to obtain the retentate, namely the agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating the agar oligosaccharide iron solution, placing the concentrated agar oligosaccharide iron solution in a vacuum freeze dryer, drying the agar oligosaccharide iron solution until the water content is below 5wt% at the absolute pressure of 20Pa in a drying chamber, the pre-freezing temperature of-35 ℃ and the resolution temperature of 35 ℃, and sieving the crushed agar oligosaccharide iron solution with a 80-mesh sieve to obtain the agar oligosaccharide iron.
In order to fully disclose the method for preparing the iron complex oligosaccharide of the present invention, the following description is given with reference to examples.
Example 1
(1) Refining gracilaria agar: preparing gracilaria agar into 5wt% glue solution with ultrapure water, heating to 90 deg.C, stirring to dissolve completely, adding polyethylene glycol 6000 and 1% NaCl clarifier 20% of the glue solution by mass, treating with ultrasonic vibrator at 40kHz for 3min, cooling to room temperature, standing for precipitation, centrifuging, collecting separated liquid, standing for precipitation, centrifuging, mixing precipitates, washing the precipitate with cold water twice, and press-filtering for dehydration to obtain refined agar;
(2) acid degradation: preparing 5wt% of glue solution from refined agar by using ultrapure water, heating to 90 ℃, stirring until the glue solution is completely dissolved, adding 1.0mol/L HCl solution into the glue solution, treating the glue solution for 90min, dropwise adding 1.0mol/L NaOH solution into the glue solution until the pH value is 7, centrifuging the glue solution for 15min at 7000 r/min, and taking supernatant fluid to obtain agar oligosaccharide hydrolysate;
(3) high-voltage pulse electric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2O and trisodium citrate dihydrate, and the agarThe mass ratio of oligosaccharide (by dry weight) to iron ion to trisodium citrate dihydrate is 7.5: 1: 7.5, HCl is adopted to adjust the pH value to 3, after heating to 50 ℃, 20kV/cm high-voltage electric field is adopted to process for 60min, and reaction liquid is obtained;
(4) membrane separation: filtering the reaction solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3000Da, collecting the permeate, and filtering the permeate by using an ultrafiltration membrane with the molecular weight cutoff of 500Da to obtain the retentate, namely the agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating the agar oligosaccharide iron solution, placing the concentrated agar oligosaccharide iron solution in a vacuum freeze dryer, drying the agar oligosaccharide iron solution until the water content is below 5wt% at the absolute pressure of 20Pa in a drying chamber, the pre-freezing temperature of-35 ℃ and the resolution temperature of 35 ℃, and sieving the crushed agar oligosaccharide iron solution with a 80-mesh sieve to obtain the agar oligosaccharide iron.
The weight average molecular weight of the agar oligosaccharide iron prepared by the example is 1252Da, wherein the iron content is 12.83%.
Example 2
(1) Refining gracilaria agar: preparing gracilaria agar into 4wt% glue solution with ultrapure water, heating to 90 deg.C, stirring to dissolve completely, adding polyethylene glycol 6000 and 1% NaCl clarifier 15% of the glue solution by mass, treating with ultrasonic vibrator at 30kHz for 5min, cooling to room temperature, standing for precipitation, centrifuging, collecting separated liquid, standing for precipitation, centrifuging, mixing precipitates, washing the precipitate with cold water twice, and press-filtering for dehydration to obtain refined agar;
(2) acid degradation: preparing refined agar into 4wt% glue solution with ultrapure water, heating to 90 deg.C, stirring to dissolve completely, adding 1.0mol/L HCl solution, treating for 90min, dropwise adding 1.0mol/L NaOH solution to pH 7, centrifuging at 7000 r/min for 15min, and collecting supernatant to obtain agar oligosaccharide hydrolysate;
(3) high-voltage pulse electric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2O and trisodium citrate dihydrate, the mass ratio of agar oligosaccharide (dry weight) to iron ion to trisodium citrate dihydrate is 5: 1: 2.5, NaOH is adopted to adjust the pH value to 5, after heating to 65 ℃, the mixture is treated for 45min by adopting a 40 kV/cm high-voltage electric fieldObtaining reaction liquid;
(4) membrane separation: filtering the reaction solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3000Da, collecting the permeate, and filtering the permeate by using an ultrafiltration membrane with the molecular weight cutoff of 500Da to obtain the retentate, namely the agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating the agar oligosaccharide iron solution, placing the concentrated agar oligosaccharide iron solution in a vacuum freeze dryer, drying the agar oligosaccharide iron solution until the water content is below 5wt% at the absolute pressure of 20Pa in a drying chamber, the pre-freezing temperature of-35 ℃ and the resolution temperature of 35 ℃, and sieving the crushed agar oligosaccharide iron solution with a 80-mesh sieve to obtain the agar oligosaccharide iron.
The weight average molecular weight of the agar oligosaccharide iron prepared by the example is 1355Da, wherein the iron content is 14.03 percent.
Example 3
(1) Refining gracilaria agar: preparing gracilaria agar into 3wt% glue solution with ultrapure water, heating to 90 deg.C, stirring to dissolve completely, adding 10% polyethylene glycol 6000 and 1% NaCl clarifier into the glue solution, treating with ultrasonic vibrator at 20kHz for 8min, cooling to room temperature, standing for precipitation, centrifuging, collecting separated liquid, standing for precipitation, centrifuging, mixing precipitates, washing the precipitate with cold water twice, and press-filtering for dehydration to obtain refined agar;
(2) acid degradation: preparing refined agar into 3wt% glue solution with ultrapure water, heating to 90 deg.C, stirring to dissolve completely, adding 1.0mol/L HCl solution, treating for 90min, dropwise adding 1.0mol/L NaOH solution to pH 7, centrifuging at 7000 r/min for 15min, and collecting supernatant to obtain agar oligosaccharide hydrolysate;
(3) high-voltage pulse electric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2O and trisodium citrate dihydrate, the mass ratio of agar oligosaccharide (by dry weight) to iron ions to trisodium citrate dihydrate is 2.5: 1: 0.83, NaOH is adopted to adjust the pH value to 7, after heating to 80 ℃, 60kV/cm high-voltage electric field is adopted for processing for 30min, and reaction liquid is obtained;
(4) membrane separation: filtering the reaction solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 3000Da, collecting the permeate, and filtering the permeate by using an ultrafiltration membrane with the molecular weight cutoff of 500Da to obtain the retentate, namely the agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating the agar oligosaccharide iron solution, placing the concentrated agar oligosaccharide iron solution in a vacuum freeze dryer, drying the agar oligosaccharide iron solution until the water content is below 5wt% at the absolute pressure of 20Pa in a drying chamber, the pre-freezing temperature of-35 ℃ and the resolution temperature of 35 ℃, and sieving the crushed agar oligosaccharide iron solution with a 80-mesh sieve to obtain the agar oligosaccharide iron.
The weight average molecular weight of the agar oligosaccharide iron prepared by the example is 1196Da, wherein the iron content is 11.26%.
Test of
60 SD rats were randomly divided into a normal control group (12) and an iron deficiency model group (48). The normal control group is fed with control feed, and the iron-deficiency model group is fed with low-iron feed. All rats were fed freely and after 28 days of feeding, when hemoglobin (Hb) was less than 70g/L, i.e. iron deficiency anemia was successfully modeled. Then the model group rats are divided into an iron-deficiency model group, a ferrous gluconate group, a ferrous sulfate group and an agar oligosacchride iron group, and each group contains 12 animals. Respectively irrigating the rats of the ferrous gluconate group and the ferrous sulfate group with gastric gluconic acid ferrous salt solution and ferrous sulfate solution, respectively feeding half dose of the agar oligosacchare iron solution to the rats of the agar oligosacchare iron group, irrigating the rats of the iron deficiency model group and the rats of the normal control group with physiological saline with the same volume as the gastric acid, and continuously irrigating the rats for 28 days by one time every day. The other groups were fed with low-iron feed except for the normal control group which was always fed with control feed. After the test was completed, fasting was performed for 12 hours, and a blood sample was collected by cardiac puncture and subjected to a blood routine test, the results of which are shown in table 1.
TABLE 1 comparison of iron supplementation effect of different iron supplements on iron deficient rats
Note: the same column of shoulder marks in lower case letters, with different letters indicating significant differences between the different groups (P < 0.05).
Hb, RBC and HCT in Table 1 represent the major indicators of blood routine-hemoglobin, red blood cells, hematocrit, respectively. Compared with ferrous gluconate and ferrous sulfate, half of agar oligosaccharide iron can achieve equivalent iron supplementing effect, so that the blood of the iron-deficient rat is recovered to normal level conventionally.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (6)
1. A preparation method of agar oligosaccharide iron is characterized by comprising the following steps:
(1) refining gracilaria agar: preparing gracilaria agar into 3-5 wt% glue solution with ultrapure water, heating to 90 ℃, stirring until the glue solution is completely dissolved, adding a clarifying agent into the glue solution, performing ultrasonic vibration treatment, cooling to room temperature, standing for precipitation, centrifuging, performing the same clarification and ultrasonic vibration treatment on a separation solution, standing for precipitation, centrifuging, combining precipitates, washing the precipitates twice with cold water, and performing filter pressing and dehydration to obtain refined agar;
(2) acid degradation: preparing refined agar into a glue solution with the concentration of 3-5 wt% by using ultrapure water, heating to 90 ℃, stirring until the refined agar is completely dissolved, adding an HCl solution with the final concentration of 1.0mol/L to treat for 90min, dropwise adding a 1.0mol/L NaOH solution until the pH value is 7, centrifuging at 7000 r/min for 15min, and taking a supernatant to obtain an agar oligosaccharide hydrolysate;
(3) high-voltage pulse electric field assisted chelation: adding FeCl into agar oligosaccharide hydrolysate3 .6H2Adjusting the pH value of O and trisodium citrate dihydrate to 3-7 by adopting HCl or NaOH, heating to 50-80 ℃, and treating by adopting a high-voltage electric field to obtain a reaction solution;
(4) membrane separation: performing secondary separation by using an ultrafiltration membrane, wherein a trapped fluid obtained by the secondary separation is an agar oligosaccharide iron solution;
(5) freeze drying and crushing: vacuum concentrating agar oligosaccharide iron solution, placing in a vacuum freeze dryer, drying at-35 deg.C under the absolute pressure of 20Pa in the drying chamber and 35 deg.C under the desorption temperature until the water content is below 5wt%, pulverizing, and sieving with 80 mesh sieve to obtain agar oligosaccharide iron;
the clarifying agent in the step (1) is composed of polyethylene glycol 6000 and NaCl, wherein the mass ratio of the polyethylene glycol 6000 to the glue solution is 10-20%.
2. The method for preparing agar oligosaccharide iron as claimed in claim 1, wherein the ultrasonic vibration treatment in step (1) is carried out at an ultrasonic frequency of 20 kHz-40 kHz for 3 min-8 min.
3. The method for preparing agar oligosaccharide iron as claimed in claim 1, wherein FeCl added in step (3)3 .6H2The proportion of the O and the trisodium citrate dihydrate is based on the dry weight of agar oligosaccharides in the agar oligosaccharide hydrolysate, namely the mass ratio of the dry weight of the agar oligosaccharides to the iron ions to the trisodium citrate dihydrate is 7.5-2.5: 1: 7.5-0.83.
4. The method for preparing agar oligosaccharide iron as claimed in claim 1, wherein the high-voltage electric field treatment in step (3) is a 20 kV/cm-60 kV/cm high-voltage electric field treatment for 30 min-60 min.
5. The method for preparing agar oligosaccharide iron as claimed in claim 1, wherein the ultrafiltration membrane in step (4) is used for the second separation, wherein an ultrafiltration membrane with a molecular weight cutoff of 3000Da is used for the first separation, a permeate is collected after a reaction solution is filtered, and the permeate is filtered by an ultrafiltration membrane with a molecular weight cutoff of 500Da to perform the second separation.
6. The method of any one of claims 1-5, wherein the iron agar oligosaccharide is obtained.
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