CN110589864A - Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent - Google Patents
Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent Download PDFInfo
- Publication number
- CN110589864A CN110589864A CN201910926246.4A CN201910926246A CN110589864A CN 110589864 A CN110589864 A CN 110589864A CN 201910926246 A CN201910926246 A CN 201910926246A CN 110589864 A CN110589864 A CN 110589864A
- Authority
- CN
- China
- Prior art keywords
- calcium carbonate
- trypsin
- lactic acid
- oyster
- nano calcium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
- A61K33/10—Carbonates; Bicarbonates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/02—Nutrients, e.g. vitamins, minerals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01F—COMPOUNDS OF THE METALS BERYLLIUM, MAGNESIUM, ALUMINIUM, CALCIUM, STRONTIUM, BARIUM, RADIUM, THORIUM, OR OF THE RARE-EARTH METALS
- C01F11/00—Compounds of calcium, strontium, or barium
- C01F11/18—Carbonates
- C01F11/181—Preparation of calcium carbonate by carbonation of aqueous solutions and characterised by control of the carbonation conditions
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01F—COMPOUNDS OF THE METALS BERYLLIUM, MAGNESIUM, ALUMINIUM, CALCIUM, STRONTIUM, BARIUM, RADIUM, THORIUM, OR OF THE RARE-EARTH METALS
- C01F11/00—Compounds of calcium, strontium, or barium
- C01F11/18—Carbonates
- C01F11/182—Preparation of calcium carbonate by carbonation of aqueous solutions and characterised by an additive other than CaCO3-seeds
- C01F11/183—Preparation of calcium carbonate by carbonation of aqueous solutions and characterised by an additive other than CaCO3-seeds the additive being an organic compound
-
- C—CHEMISTRY; METALLURGY
- C01—INORGANIC CHEMISTRY
- C01P—INDEXING SCHEME RELATING TO STRUCTURAL AND PHYSICAL ASPECTS OF SOLID INORGANIC COMPOUNDS
- C01P2004/00—Particle morphology
- C01P2004/30—Particle morphology extending in three dimensions
- C01P2004/32—Spheres
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Nanotechnology (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Crystallography & Structural Chemistry (AREA)
- Veterinary Medicine (AREA)
- Geology (AREA)
- Animal Behavior & Ethology (AREA)
- Physics & Mathematics (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Physics & Mathematics (AREA)
- Epidemiology (AREA)
- Hematology (AREA)
- Composite Materials (AREA)
- Manufacturing & Machinery (AREA)
- Nutrition Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Diabetes (AREA)
- Materials Engineering (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent, which comprises the steps of dissolving oyster shell powder by using lactic acid, filtering and concentrating a dissolved solution, adjusting the pH of the solution to 11 ~ 12 by using ammonia water, then adding trypsin, and introducing CO2And (5) gas is generated to obtain the nano calcium carbonate. The invention not only solves the problem of serious environmental pollution caused by the waste oyster shells, but also changes waste into valuable. Compared with calcium carbonate originally contained in oyster shell, the nano calcium carbonate of the present invention has smaller particle, better crystal form, better absorption, high bioavailability and capacity of eliminating calcium carbonateRemoving the original fishy smell of oyster shell, and improving taste. Compared with the current mainstream production process, the production process disclosed by the invention is simple to operate, green and environment-friendly, does not produce secondary pollution, saves the production cost, and has a good popularization and application prospect.
Description
Technical Field
The invention relates to the technical field of preparation of calcium preparations, in particular to a method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent.
Background
With the improvement of living standard of people, the aquaculture industry is rapidly developed, and under the large environment, the oyster cultivation industry and the oyster processing industry are rapidly expanded. In shellfish seafood, oyster is popular with consumers because of its delicious taste and rich nutrition. However, at the same time, the shells that cannot be eaten are discarded as garbage, resulting in a large amount of oyster shells becoming waste or a low-value resource. The waste oyster shells not only occupy certain beach and land resources, but also can be rotten and smelly to cause environmental pollution. Therefore, how to fully and comprehensively develop and utilize oyster shells and turn the oyster shells into wealth is a major problem to be solved urgently.
Calcium is an essential mineral element for the human body and is an important component constituting bones and teeth of the body. Research shows that calcium is widely distributed in muscles, blood and tissues and organs of human bodies and plays an extremely important role in cell division, heart rate and blood pressure maintenance and cell membrane integrity maintenance. In addition, calcium can be involved in activities such as nerve and muscle activity, hormone secretion, blood coagulation, cell adhesion, etc., and is also an activator of many enzymes in the metabolic process of the body. The lack of calcium in infants can affect the growth and development of infants in the aspects of movement, speech expression, intelligence, body shape, etc. Rickets are easily produced by calcium deficiency of teenagers. Middle-aged and elderly people are easy to suffer from arteriosclerosis, hypertension and other diseases due to calcium deficiency. Although the social living standard is continuously improved and more nutrient substances are taken, the phenomenon of calcium deficiency of people in China is still relatively serious.
Among calcium supplement preparations used at home and abroad, nano calcium carbonate is more and more favored by people. The nano calcium carbonate has smaller particle size, has quantum size effect, small size effect, surface effect and macroscopic quantum tunneling effect which are not possessed by common calcium carbonate, successfully solves the problems of dissolution speed and absorption rate of calcium preparations, and further increases the bioavailability of the calcium preparations. In recent years, nano calcium carbonate plays more and more important roles in the pharmaceutical and food industries, gradually replaces common calcium and even organic calcium, and becomes an important calcium source additive. The oyster shell calcium carbonate content is up to more than 95 percent, is a natural calcium source produced by organisms, and is a precious resource for preparing nano calcium carbonate.
At present, the production process for preparing the nano calcium carbonate by utilizing the oyster shells adopts two methods of roasting or hydrochloric acid dissolution. But the roasting process needs high temperature conditions, which greatly increases the energy consumption of the production process. In order to overcome the defects of the procedures, the double decomposition reaction process for preparing the nano calcium carbonate by dissolving oyster shells with hydrochloric acid and ammonium carbonate is studied, but the use of the hydrochloric acid can cause a large amount of Cl to be adsorbed on the nano calcium carbonate-The production of the catalyst requires a lot of time and washing water to remove the Cl-The production cost of the product is very high, and large-scale industrial production is not utilized. In addition, the crystal forms of the nano calcium carbonate prepared at present are limited, and the production control difficulty of the crystal forms is high, so that a crystal form control agent needs to be added to control the growth speed of the crystal face, the crystal size and the crystal form of the nano calcium carbonate.
Trypsin is a proteolytic enzyme which is present in the animal pancreas as an inactive proenzyme, in Ca2+Is activated by enterokinase or active trypsin itself and is converted into active trypsin. The trypsin has the functions of resisting inflammation, accelerating wound surface purification, promoting granulation tissue regeneration and the like. It is clinically used for empyema, local edema and abscess caused by surgical inflammation and ulcer, etc. Therefore, as an important tool enzyme in production and life, the trypsin is widely applied in the field of medical research.
Disclosure of Invention
The invention aims to provide a method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent comprises the following steps:
1) dissolving oyster shell with lactic acid, extracting calcium ion from oyster shell, and filtering lactic acid solution to obtain filtrate;
2) adjusting the pH value of the filtrate to 11-12 by using ammonia water, then adding trypsin into the filtrate, and introducing CO after complete dissolution2Gas, water bath magnetic stirring, collecting crystals after ventilation, washing and drying the crystals to obtain the nano calcium carbonate.
Further, the step 1) is specifically as follows: cleaning oyster shell, removing impurities, crushing, and sieving with a 100-mesh sieve to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, and simultaneously placing the beaker in a water bath kettle to react under the condition of magnetic stirring, wherein the reaction conditions are as follows: the concentration of the lactic acid is 1.75-2.25 mol/L, the temperature is 50-60 ℃, the reaction time is 25-35 min, the magnetic stirring speed is 250-350 rpm, after the lactic acid completely dissolves the oyster shell powder, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain the filtrate.
Preferably, the reaction conditions are as follows: the concentration of lactic acid is 2mol/L, the temperature is 50 ℃, the reaction time is 30min, and the magnetic stirring speed is 350 rpm.
Step 2) adding the trypsin in an amount which satisfies the following conditions: and (3) A and B are 0.4065: 0.0041-0.8130, wherein A is the mass of calcium in the filtrate, and B is the mass of trypsin. Furthermore, A and B are 0.4065: 0.0203-0.061.
Step 2) introduction of CO2The flow rate of the gas was 1L/min.
The temperature of the water bath in the step 2) is 23-27 ℃.
The nano calcium carbonate prepared by the invention can be applied to the preparation of calcium supplement preparations.
The invention adopts lactic acid to dissolve oyster shell powder, the dissolved solution is filtered and concentrated, then trypsin is added, and CO is introduced2Gas to prepare the nano calcium carbonate, and has the following advantages:
1. the invention utilizes the lactic acid to dissolve calcium ions in the oyster shells, compared with other organic acids such as hydrochloric acid and the like, the lactic acid not only can not generate side effect on human bodies after entering the human bodies, but also can be reused by the human bodies after releasing the calcium ions. Because it is not necessary toTo remove Cl adsorbed on the surface of calcium carbonate-Therefore, the production process is simple to operate, green and environment-friendly, does not produce secondary pollution, saves the production cost and has good popularization and application prospects.
2. The invention utilizes trypsin as a crystal form control agent. At present, researches on the influence of crystal form control agents such as inorganic salts, organic alcohols, proteins and amino acids on the nucleation of nano calcium carbonate are reported, but researches on the use of trypsin as the crystal form control agent are not reported.
3. Compared with the original calcium carbonate in the oyster shell, the nano calcium carbonate prepared by the invention has the characteristics of smaller particles, better crystal form, better absorption, high bioavailability and the like, and can also remove the original fishy smell of the oyster shell and improve the mouthfeel.
Drawings
FIG. 1 shows the introduction of CO into the lactic acid solution2Infrared spectrum of the post product;
FIG. 2 shows the introduction of CO into the calcium chloride solution2Infrared spectrum of the post product;
FIG. 3 shows the introduction of CO into the lactic acid solution2Thermogravimetric curve of the post product;
FIG. 4 shows Ca in the filtrate at different reaction temperatures2+Content (c);
FIG. 5 shows Ca in the filtrate at different concentrations of lactic acid2+Content (c);
FIG. 6 shows Ca in the filtrate at different reaction times2+Content (c);
FIG. 7 shows Ca in the filtrate at different stirring speeds2+Content (c);
FIG. 8 shows the morphology of calcium carbonate without trypsin;
FIG. 9 shows the morphology of calcium carbonate with 0.0041g trypsin added;
FIG. 10 shows the morphology of calcium carbonate with 0.0203g trypsin added;
FIG. 11 shows the morphology of calcium carbonate with the addition of 0.0610g of trypsin;
FIG. 12 shows the morphology of calcium carbonate with 0.8130g of trypsin.
Detailed Description
The substance and advantageous effects of the present invention will be described in further detail with reference to examples, which are provided only for illustrating the present invention and not for limiting the present invention. In addition, after reading the teaching of the present invention, those skilled in the art can make various changes or modifications to the invention, and these equivalents also fall within the scope of the claims appended to the present application.
EXAMPLE 1 lactic acid dissolution of oyster Shell to prepare calcium carbonate
The collected oyster shells are washed clean by tap water, and the sand and other impurities attached to the oyster shells are brushed by a brush and are hammered for standby. Pulverizing processed Concha Ostreae into powder with universal pulverizer, sieving with 100 mesh sieve, weighing, and bottling.
10g of oyster shell powder was weighed into a 500mL beaker, 216mL of a lactic acid (1mol/L) solution was slowly poured into the beaker, and at the same time, the beaker was placed in a 30 ℃ water bath and a stirrer was added to the beaker to adjust the rotation speed to 300rpm, and the time for the lactic acid to completely dissolve the oyster shell powder was recorded. After the lactic acid completely dissolves the oyster shell powder, closing the water bath, filtering the lactic acid dissolved solution, measuring the volume of the filtrate, and recording as VGeneral assemblymL, the filtrate is stored for later use.
The EDTA method for determining the content of calcium ions in the lactic acid dissolved oyster shell filtrate comprises the following specific operations: taking 1mL of filtrate obtained by dissolving oyster shell powder in lactic acid by using a 1mL pipette, diluting by a certain multiple, taking 25mL of diluent by using a pipette, adjusting the pH value of the diluent to be 12-13 by using 6 mol/L NaOH, transferring the diluent to a conical flask, adding a proper amount of calcium carboxylic acid indicator, fully shaking up, titrating the solution by using 0.000625mol/L EDTA solution, continuously oscillating the conical flask while titrating, changing the solution from rose red to brilliant blue within 30s without fading to indicate that the titration end point is reached, recording the reading of a titration tube, carrying out parallel titration twice, calculating the average value of the consumed EDTA solution, and recording the average value as V1And mL, calculating the content of calcium ions in the filtrate.
The calculation formula of calcium ions contained in the filtrate is as follows:
WCa2+＝C×(V1/1000)×M×Vgeneral assemblyX (dilution volume/25)/(10X 90%. times.40%)
Wherein C is the concentration of EDTA: 0.000625 mol/L;
m is calcium atomic mass: 40.078 g/mol.
In the above experiment VGeneral assembly208mL, 16.20mL EDTA consumed, and 93.78% calcium ion in the filtrate.
Taking a filtrate obtained by dissolving oyster shell powder with lactic acid, adjusting the pH value of the filtrate to 11-12 with ammonia water, and introducing CO2Collecting the precipitate, washing, drying, taking a little of the sample to measure the infrared spectrum, and the specific operation process is as follows: 100-200 mg of KBr powder is put into an agate mortar, and about 2mg of a crystal sample baked to constant weight is added and fully ground. After grinding, the powder is transferred to a proper mould to be evenly distributed, and tabletting is carried out on a manual tablet press to form transparent sheets. Taking a KBr blank tablet as a reference, and taking the KBr blank tablet as a reference, wherein the KBr blank tablet has a wavelength of 4000-400 cm-1And (4) internally scanning infrared spectrum, and analyzing and identifying characteristic functional groups of the product.
The results of the infrared spectroscopy are shown in FIG. 1. As can be seen, 1460cm in the infrared spectrum-1Nearby peak and 875cm-1The sharp absorption peak of (2) is all infrared absorption peaks of calcium carbonate. At 1459cm-1Has C-O antisymmetric telescopic vibration of 876 cm-1The appearance of CO3 2-Out-of-plane deformation peak at 724cm-1An in-plane deformation vibration peak of 0-C-O appears. The infrared spectrum is similar to that of calcium carbonate, and can be used for deducing that the lactic acid dissolved solution is introduced with CO after the pH is adjusted2And then converted into calcium carbonate.
FIG. 2 shows the introduction of CO into the calcium chloride solution2The infrared spectrum of the product is 1420cm-1、875cm-1The sharp absorption peak of (2) is all infrared absorption peaks of calcium carbonate. 1420cm-1The peak appeared is C-O reverse stretching vibration, 875cm-1Has a peak of CO3 2-Out-of-plane bending vibration peak at 710cm-1The peaks appearing on the left and right are the O-C-O in-plane bending vibration peaks. Introducing CO into the spectrogram and the lactic acid solution2The spectrogram is similar, so that the pH of the lactic acid solution is adjusted and CO is introduced2And then converted into calcium carbonate.
And (3) carrying out thermogravimetric analysis on the dried sample, wherein the specific operations are as follows: the sample was dried to constant weight and the product was analyzed using a thermogravimetric analyzer. And (3) measuring the thermogravimetric curve of the sample, preheating the instrument, setting parameters such as temperature, heating rate and the like on experimental software, putting the sample into a crucible, and operating the experimental software. The decomposition temperature of each sample was measured.
The thermogravimetric results are shown in FIG. 3, where the sample began to decompose at around 650 deg.C, with an approximate weight loss of 44%, i.e., the decomposition of calcium carbonate to CaO and CO2。
EXAMPLE 2 Single factor assay to determine optimal factor levels for lactic acid solubilization of oyster Shell
The influence of the dissolution rate of the calcium ions in the oyster shells of the lactic acid under different acid concentrations, temperatures, reaction time lengths and stirring speeds is examined, and the optimal level of the lactic acid under various factors is determined by taking the high and low calcium lactate yield as an index.
2-1 temperature influence on dissolution rate of calcium ions in oyster shell
10g of oyster shell powder is taken and placed in a 500mL beaker, and the oyster shell powder is dissolved at 25 ℃, 35 ℃, 45 ℃, 55 ℃, 65 ℃ and 75 ℃ respectively under the condition that the concentration of lactic acid, the reaction time and the stirring speed are the same. After complete dissolution, filtration was carried out and the volume of the filtrate was measured. Taking 1mL of filtrate, diluting by 1000 times, taking 25mL of diluent for titration, recording the volume of the EDTA consumption solution, carrying out parallel titration twice, calculating the average value of the EDTA consumption solution, calculating the calcium ion elution amount according to the calculation formula of calcium ions in example 1, and determining the optimal temperature for dissolving the oyster shell powder.
The results are shown in FIG. 4: the calcium content in oyster shell is about 40%, the calcium dissolution content is gradually increased when the temperature is gradually increased, the calcium dissolution content is maximum when the temperature reaches 65 ℃, and the calcium dissolution content is reduced when the temperature is continuously increased. When the temperature is 55 ℃, the calcium dissolution content is close to the maximum value, and in order to save resources, the preferred temperature is 55 ℃.
Influence of 2-2 lactic acid concentration on dissolution rate of calcium ions in oyster shell
10g of oyster shell powder is respectively taken and placed in a 500mL beaker, the stirring speed is kept the same under the optimal temperature and the optimal reaction time, and the concentration of lactic acid is respectively controlled to be 0.5mol/L, 1.0mol/L, 1.5mol/L, 2.0mol/L, 2.5mol/L and 3.0mol/L to dissolve the oyster shell powder. After complete dissolution, filtration was carried out and the volume of the filtrate was measured. Taking 1mL of filtrate, diluting by 1000 times, taking 25mL of diluent for titration, recording the volume of the EDTA consumption solution, carrying out parallel titration twice, calculating the average value of the EDTA consumption solution, calculating the calcium ion dissolution amount according to the calculation formula of calcium ions in the embodiment 1, and determining the better concentration for dissolving the oyster shell powder.
The results are shown in FIG. 5: the calcium content in oyster shell is about 40%, when the lactic acid concentration is gradually increased, the calcium dissolved content is gradually increased, when the lactic acid concentration is 2mol/L, the calcium dissolved content is maximum, the lactic acid concentration is continuously increased, and the calcium dissolved content is reduced, so 2mol/L is taken as the better concentration.
2-3 influence of reaction duration on dissolution rate of calcium ions in oyster shell
10g of oyster shell powder is respectively put into a 500mL beaker, the concentration of lactic acid and the stirring speed are kept the same at the optimal temperature, and the reaction time is respectively controlled to be 15min, 30min, 60min, 90min, 120min and 150min to dissolve the oyster shell powder. After complete dissolution, filtration was carried out and the volume of the filtrate was measured. Taking 1mL of filtrate, diluting by 1000 times, taking 25mL of diluent for titration, recording the volume of the EDTA consumption solution, carrying out parallel titration twice, calculating the average value of the EDTA consumption solution, calculating the calcium ion elution amount according to the calculation formula of calcium ions in the embodiment 1, comparing the calcium ion elution amounts at various reaction times, and determining the better reaction time for dissolving the oyster shell powder.
The experimental results are shown in fig. 6: the calcium content in oyster shell is about 40%, when the reaction time is gradually increased, the calcium dissolved content is gradually increased, when the reaction time is 30min, the calcium dissolved content is maximum, and when the reaction time is continuously increased, the calcium dissolved content tends to be stable, so that 30min is selected as a preferable reaction time.
2-4 stirring speed influences the dissolution of calcium ions in oyster shells
10g of oyster shell powder is taken and placed in a 500mL beaker, and the oyster shell powder is dissolved by respectively controlling the stirring speed to be 0, 100rpm, 200rpm, 300rpm, 400rpm and 500rpm under the conditions of optimal temperature, optimal lactic acid concentration and optimal reaction time. After complete dissolution, filtration was carried out and the volume of the filtrate was measured. Taking 1mL of filtrate, diluting by 1000 times, taking 25mL of diluent for titration, recording the volume of the EDTA consumption solution, carrying out parallel titration twice, calculating the average value of the EDTA consumption solution, and calculating the calcium ion elution amount according to the calculation formula of calcium ions in the embodiment 1 to determine the better stirring speed for dissolving the oyster shell powder.
The results of the experiment are shown in FIG. 7: the calcium content in oyster shell is about 40%, the calcium dissolution content is gradually increased when the stirring speed is gradually increased, the calcium dissolution content is maximum when the stirring speed is 300rpm, and the calcium dissolution amount is reduced when the stirring speed is continuously increased, so that the magnetic stirring speed of 300rpm is used as the preferable stirring speed.
In summary, the following steps: the single factor test result shows that when the calcium ions in the oyster shells are dissolved by the lactic acid, the preferable temperature of the reaction is 55 ℃, the preferable reaction time is 30min, the preferable concentration of the lactic acid is 2mol/L, and the preferable stirring speed is 300 rpm.
Example 3 determination of optimum production Process for dissolving calcium ion in oyster Shell by lactic acid through orthogonal experiment
According to the preferred reaction conditions determined by the one-factor method, an orthogonal experiment is carried out by four factors of A (lactic acid concentration/mol/L), B (temperature/DEG C), C (reaction time/min) and D (stirring speed/rpm), each factor selects three levels near the preferred conditions, and L is referred to9(34) Orthogonal tables experiments were carried out (Table 1), with Ca2+The dissolution rate of the oyster shell calcium is an investigation index, the influence of various factors on the dissolution rate of the oyster shell calcium is investigated, the optimal factor level combination is determined, and the optimal process conditions for dissolving the calcium ions in the oyster shell are determined.
TABLE 1 orthogonal Experimental Table
According to the single-factor experimental result, the main factors influencing the calcium dissolution in the process of dissolving the calcium ions in the oyster shell by the lactic acid comprise the lactic acid concentration, the temperature, the reaction time and the stirring speed. Under the same experimental conditions as the single factor method, changing different lactic acid concentrations, temperatures, reaction durations and stirring speeds, and collecting oyster shell powder10g, adding 200mL of lactic acid with different concentrations under each test condition, and measuring the calcium content of the oyster shell dissolved by lactic acid in each group under the same other conditions. According to the quadrature L9(34) An orthogonal experimental scheme is designed, the optimal process conditions for dissolving the calcium ions in the oyster shells by the lactic acid are investigated according to the experimental scheme, and the experimental results are shown in table 2. Of the 4 factors listed, the 2 nd level of lactic acid concentration is most preferred, the 1 st level of temperature is most preferred, the 2 nd level of reaction time length is most preferred, and the 3 rd level of stirring speed is most preferred. Therefore, the optimal reaction conditions for extracting calcium ions from oyster shells by lactic acid are as follows: the concentration of lactic acid is 2mol/L, the temperature is 50 ℃, the reaction time is 30min, the stirring speed is 350rpm, and under the optimal condition, the dissolution rate of calcium ions in the oyster shells dissolved by lactic acid can reach 39.48 percent. By comparing the R values, the influence of four factors on the calcium dissolution rate is shown as follows: stirring speed>Concentration of lactic acid>Temperature of>The reaction time is long.
TABLE 2 calcium content dissolved by reaction of lactic acid with oyster Shell under different reaction conditions
EXAMPLE 4 preparation of Nano calcium carbonate by adding Trypsin (0g)
Under the optimal process conditions obtained in the orthogonal experiment, namely, the concentration of lactic acid is 2mol/L, the reaction temperature is 50 ℃, the reaction time is 30min, the stirring speed is 350rpm, oyster shell powder is dissolved, the filtrate is concentrated, the volume after concentration is about 37mL, the pH of the solution is adjusted to 11.76 by ammonia water (about 250mL of consumed ammonia water), and at this time, the total volume of the solution is 269mL, and the solution is stored for later use. After 1mL of the solution was diluted to 500mL, 25mL of the solution was titrated with EDTA to give an EDTA solution having a concentration of 0.000625mol/L, and the concentration of the EDTA solution was calculated to be 0.33875mol/L by performing two parallel measurements.
Taking 30mL of the solution in a beaker, wherein the calcium content is 0.4065g, adding 0g of trypsin, after complete dissolution, introducing CO at the flow rate of 1L/min2Gas, water bath magnetic stirring is carried out at 25 ℃. After the end of the aeration, the filtrate and the calcium carbonate crystals were collected. Taking 2mL of the filtrate, and measuring calcium ionsAnd (4) concentration. Introducing CO through the solution2The conversion of calcium ions was calculated to be 99.20% from the change in calcium ion concentration before and after carbonization. And (5) fully washing the crystals, drying the crystals to constant weight by using a constant-temperature air drying oven, and weighing the crystals.
The morphology characteristics of the calcium carbonate are observed by adopting a Scanning Electron Microscope (SEM), and the specific operation is as follows: cutting the conductive adhesive into a proper size, directly adhering the conductive adhesive on a copper sheet, directly scattering a sample to be detected on the conductive adhesive by means of foreign matters, and slightly blowing off the sample by using an ear washing ball to remove the sample which is not adhered on the conductive adhesive. And then, injecting a sample, and observing the appearance, the crystal form and the particle size of each sample under different times of amplification.
SEM images of calcium carbonate without trypsin are shown in fig. 8. a. And b, c and d are images which are magnified by 1000, 5000, 10000 and 20000 times under a scanning electron microscope respectively, and it can be seen from the images that calcium carbonate without any additive is in an aggregation state, the crystal of the calcium carbonate is in a vaterite shape, a plurality of tiny particles are arranged on the surface of a sphere, the tiny particles in nanometer scale are aggregated to form microspheres with the diameter of about 10 mu m, and the tiny microspheres with the diameter of 5-6 mu m are attached. The particle size is large and non-uniform.
EXAMPLE 5 preparation of Nano calcium carbonate by adding Trypsin (0.0041g)
Taking 30mL of the solution in a beaker, wherein the calcium content is 0.4065g, adding 0.0041g of trypsin, after complete dissolution, introducing CO at the flow rate of 1L/min2Gas, water bath magnetic stirring is carried out at 25 ℃. After the end of the aeration, the filtrate and the calcium carbonate crystals were collected. 2mL of the filtrate was taken, and the calcium ion concentration was measured. Introducing CO through the solution2The conversion rate of calcium ions was calculated to be 99.17% from the change in calcium ion concentration before and after carbonization. And (5) fully washing the crystals, drying the crystals to constant weight by using a constant-temperature air drying oven, and weighing the crystals.
The morphology of calcium carbonate was observed by SEM, and the procedure was the same as in example 4. An SEM image of calcium carbonate with 0.0041g trypsin added is shown in FIG. 9. As can be seen from the figure, the crystals are still in the form of vaterite, but the particle size is reduced, a small part of microspheres is about 10 μm, the majority of microspheres are small microspheres of 5-6 μm, and the particle size is still nonuniform.
EXAMPLE 6 preparation of Nano calcium carbonate by adding Trypsin (0.0203g)
Taking 30mL of the solution in a beaker, wherein the calcium content is 0.4065g, adding 0.0203g of trypsin, after complete dissolution, introducing CO at the flow rate of 1L/min2Gas, water bath magnetic stirring is carried out at 25 ℃. After the end of the aeration, the filtrate and the calcium carbonate crystals were collected. 2mL of the filtrate was taken, and calcium ion was measured. Introducing CO through the solution2The conversion rate of calcium ions was calculated to be 99.32% from the change in calcium ion concentration before and after carbonization. And (5) fully washing the crystals, drying the crystals to constant weight by using a constant-temperature air drying oven, and weighing the crystals.
The morphology of calcium carbonate was observed by SEM, and the procedure was the same as in example 4. An SEM image of calcium carbonate with 0.0203g trypsin added is shown in FIG. 10. As can be seen from the figure, the crystals are still in the form of vaterite, but the particle size gradually changes to 2-4 μm, and the particle size is more uniform.
EXAMPLE 7 preparation of Nano calcium carbonate by adding Trypsin (0.0610g)
Taking 30mL of solution in a beaker, wherein the calcium content is 0.4065g, adding 0.0610g of trypsin, after complete dissolution, introducing CO at the flow rate of 1L/min2Gas, water bath magnetic stirring is carried out at 25 ℃. After the end of the aeration, the filtrate and the calcium carbonate crystals were collected. 2mL of the filtrate was taken, and calcium ion was measured. Introducing CO through the solution2The conversion rate of calcium ions was calculated to be 99.26% from the change in calcium ion concentration before and after carbonization. And (5) fully washing the crystals, drying the crystals to constant weight by using a constant-temperature air drying oven, and weighing the crystals.
The morphology of calcium carbonate was observed by SEM, and the procedure was the same as in example 4. An SEM image of calcium carbonate with the addition of 0.0610g trypsin is shown in FIG. 11. As can be seen from the figure, the crystals are still in the form of vaterite, but the particle size gradually changes to 2-3 μm and is uniform.
EXAMPLE 8 preparation of Nano calcium carbonate by adding Trypsin (0.8130g)
Taking 30mL of solution in a beaker, wherein the calcium content is 0.4065g, adding 0.8130g of trypsin, after complete dissolution, introducing CO at the flow rate of 1L/min2Gas, water bath magnetic stirring is carried out at 25 ℃. After the end of the aeration, the filtrate and the calcium carbonate crystals were collected. Get onThe filtrate was 2mL, and calcium ion was measured. Introducing CO through the solution2The conversion rate of calcium ions was calculated to be 99.27% from the change in calcium ion concentration before and after carbonization. And (5) fully washing the crystals, drying the crystals to constant weight by using a constant-temperature air drying oven, and weighing the crystals.
The morphology of calcium carbonate was observed by SEM, and the procedure was the same as in example 4. An SEM image of calcium carbonate with the addition of 0.8130g trypsin is shown in FIG. 12. As can be seen from the figure, the crystals are still in the form of vaterite, but the particle size gradually changes to 300-400 nm, and the particle size is uniform.
Example 9
A method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent comprises the following steps:
1) cleaning oyster shell, removing impurities, crushing, and sieving with a 100-mesh sieve to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, and simultaneously placing the beaker in a water bath kettle to react under the condition of magnetic stirring, wherein the reaction conditions are as follows: the concentration of lactic acid is 2mol/L, the temperature is 50 ℃, the reaction time is 30min, the magnetic stirring rotating speed is 350rpm, after the oyster shell powder is completely dissolved by the lactic acid, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain filtrate;
2) adjusting pH of the filtrate to 11.5 with ammonia water, adding trypsin into the filtrate, dissolving completely, and introducing CO at a flow rate of 1L/min2And (3) carrying out water bath magnetic stirring on the gas at 25 ℃, collecting crystals after the ventilation is finished, and washing and drying the crystals to obtain the nano calcium carbonate.
Wherein, the addition amount of the trypsin satisfies the following conditions: a and B are 0.4065: 0.02, A is the mass of calcium in the filtrate, and B is the mass of trypsin.
Example 10
A method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent comprises the following steps:
1) cleaning oyster shell, removing impurities, crushing, and sieving with a 100-mesh sieve to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, and simultaneously placing the beaker in a water bath kettle to react under the condition of magnetic stirring, wherein the reaction conditions are as follows: the concentration of lactic acid is 1.75mol/L, the temperature is 60 ℃, the reaction time is 25min, the magnetic stirring speed is 250rpm, after the oyster shell powder is completely dissolved by the lactic acid, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain filtrate;
2) adjusting pH of the filtrate to 11 with ammonia water, adding trypsin into the filtrate, dissolving completely, and introducing CO at a flow rate of 1L/min2And (3) carrying out water bath magnetic stirring on the gas at 23 ℃, collecting crystals after the gas introduction is finished, and washing and drying the crystals to obtain the nano calcium carbonate.
Wherein, the addition amount of the trypsin satisfies the following conditions: a and B are 0.4065: 0.04, A is the mass of calcium in the filtrate, and B is the mass of trypsin.
Example 11
A method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent comprises the following steps:
1) cleaning oyster shell, removing impurities, crushing, and sieving with a 100-mesh sieve to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, and simultaneously placing the beaker in a water bath kettle to react under the condition of magnetic stirring, wherein the reaction conditions are as follows: the concentration of lactic acid is 2.25mol/L, the temperature is 50 ℃, the reaction time is 35min, the magnetic stirring speed is 300rpm, after the oyster shell powder is completely dissolved by the lactic acid, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain filtrate;
2) adjusting pH of the filtrate to 12 with ammonia water, adding trypsin into the filtrate, dissolving completely, and introducing CO at a flow rate of 1L/min2And (3) carrying out water bath magnetic stirring on the gas at the temperature of 27 ℃, collecting crystals after the gas introduction is finished, and washing and drying the crystals to obtain the nano calcium carbonate.
Wherein, the addition amount of the trypsin satisfies the following conditions: a and B are 0.4065: 0.08, A is the mass of calcium in the filtrate, and B is the mass of trypsin.
Example 12
A method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent comprises the following steps:
1) cleaning oyster shell, removing impurities, crushing, and sieving with a 100-mesh sieve to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, and simultaneously placing the beaker in a water bath kettle to react under the condition of magnetic stirring, wherein the reaction conditions are as follows: the concentration of lactic acid is 2mol/L, the temperature is 55 ℃, the reaction time is 30min, the magnetic stirring rotating speed is 350rpm, after the oyster shell powder is completely dissolved by the lactic acid, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain filtrate;
2) adjusting the pH value of the filtrate to 11-12 by using ammonia water, then adding trypsin into the filtrate, and introducing CO at the flow rate of 1L/min after complete dissolution2And (3) carrying out water bath magnetic stirring on the gas at 25 ℃, collecting crystals after the ventilation is finished, and washing and drying the crystals to obtain the nano calcium carbonate.
Wherein, the addition amount of the trypsin satisfies the following conditions: a and B are 0.4065: 0.25, wherein A is the mass of calcium in the filtrate, and B is the mass of trypsin.
Claims (10)
1. A method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent is characterized by comprising the following steps: which comprises the following steps:
1) dissolving oyster shell with lactic acid, extracting calcium ion from oyster shell, and filtering lactic acid solution to obtain filtrate;
2) adjusting pH of the filtrate to 11 ~ 12 with ammonia water, adding trypsin into the filtrate, dissolving completely, and introducing CO2Gas, water bath magnetic stirring, collecting crystals after ventilation, washing and drying the crystals to obtain the nano calcium carbonate.
2. The method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent according to claim 1, which comprises the following steps: the step 1) is as follows: cleaning oyster shell, removing impurities, pulverizing, sieving to obtain oyster shell powder, placing the oyster shell powder in a beaker, adding a lactic acid solution, simultaneously placing the beaker in a water bath kettle, reacting under the condition of magnetic stirring, closing the water bath kettle after the lactic acid completely dissolves the oyster shell powder, and filtering the lactic acid dissolved solution to obtain a filtrate.
3. The method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent according to claim 2, which comprises the following steps: the mesh number of the screen used for crushing and sieving is 100 meshes.
4. The method for preparing oyster nano calcium carbonate by using trypsin as a crystal form control agent according to claim 2, wherein the reaction conditions comprise that the concentration of lactic acid is 1.75mol/L ~ 2.25.25 mol/L, the temperature is 50 ℃ and ~ 60 ℃, the reaction time is 25min and ~ 35min, the magnetic stirring speed is 250rpm and ~ 350rpm, after the lactic acid completely dissolves oyster shell powder, the water bath kettle is closed, and the lactic acid dissolved solution is filtered to obtain filtrate.
5. The method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent according to claim 4, which comprises the following steps: the reaction conditions were as follows: the concentration of lactic acid is 2mol/L, the temperature is 50 ℃, the reaction time is 30min, and the magnetic stirring speed is 350 rpm.
6. The method for preparing oyster nano calcium carbonate by using trypsin as a crystal form control agent according to claim 1, wherein the trypsin in the step 2) is added in an amount of A: B = 0.4065: 0.0041 ~ 0.8130, A is the mass of calcium in the filtrate, and B is the mass of trypsin.
7. The method for preparing oyster nano calcium carbonate by using trypsin as a crystal form control agent according to claim 6, wherein A: B = 0.4065: 0.0203 ~ 0.061.061.
8. The method for preparing oyster nano calcium carbonate by taking trypsin as a crystal form control agent according to claim 1, which comprises the following steps: step 2) introduction of CO2The flow rate of the gas was 1L/min.
9. The method for preparing oyster nano calcium carbonate by using trypsin as the crystal form control agent according to claim 1, wherein the water bath temperature in the step 2) is 23 ℃ and ~ 27 ℃.
10. Use of the nano calcium carbonate obtained according to any one of claims 1 to 9 in a calcium supplement formulation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910926246.4A CN110589864A (en) | 2019-09-27 | 2019-09-27 | Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910926246.4A CN110589864A (en) | 2019-09-27 | 2019-09-27 | Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110589864A true CN110589864A (en) | 2019-12-20 |
Family
ID=68864266
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910926246.4A Pending CN110589864A (en) | 2019-09-27 | 2019-09-27 | Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110589864A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS58161922A (en) * | 1982-03-18 | 1983-09-26 | Kyushiro Shirasaka | Manufacture of high purity calcium |
| CN1139073A (en) * | 1995-06-29 | 1997-01-01 | 刘昌焕 | High-efficiency calcium-replenishing agent produced with high purity calcium carbonate and its prodn process |
| CN101440031A (en) * | 2008-11-26 | 2009-05-27 | 青岛大学 | Method for preparing high-purity calcium lactate from oyster shell |
| CN103274439A (en) * | 2013-07-01 | 2013-09-04 | 广东海洋大学 | Method for preparing nano calcium carbonate by regeneratively cycling oyster shell |
| CN108217708A (en) * | 2017-12-20 | 2018-06-29 | 五邑大学 | A kind of methods and applications that nano biological calcium carbonate is prepared using waste shell as raw material |
-
2019
- 2019-09-27 CN CN201910926246.4A patent/CN110589864A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS58161922A (en) * | 1982-03-18 | 1983-09-26 | Kyushiro Shirasaka | Manufacture of high purity calcium |
| CN1139073A (en) * | 1995-06-29 | 1997-01-01 | 刘昌焕 | High-efficiency calcium-replenishing agent produced with high purity calcium carbonate and its prodn process |
| CN101440031A (en) * | 2008-11-26 | 2009-05-27 | 青岛大学 | Method for preparing high-purity calcium lactate from oyster shell |
| CN103274439A (en) * | 2013-07-01 | 2013-09-04 | 广东海洋大学 | Method for preparing nano calcium carbonate by regeneratively cycling oyster shell |
| CN108217708A (en) * | 2017-12-20 | 2018-06-29 | 五邑大学 | A kind of methods and applications that nano biological calcium carbonate is prepared using waste shell as raw material |
Non-Patent Citations (2)
| Title |
|---|
| 陈俊豪，陈贵卿著: "《珍珠与贝类的晶体结构和应用探索》", 31 December 2018 * |
| 马晓明等: ""醇水体系中胰蛋白酶调控下碳酸钙的仿生合成"", 《河南师范大学学报(自然科学版)》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107983300A (en) | Biological carbon composite of manganese dioxide modification and its preparation method and application | |
| CN110327882A (en) | The preparation method and application of multidigit activation and modified reed-south reed charcoal | |
| CN109529767A (en) | The preparation method and application method of the compound magnetic modification biological Carbon Materials of chitosan | |
| CN105622960B (en) | A kind of carboxymethyl chitosan/oxidized graphene composite aquogel and its preparation and application | |
| CN104888705B (en) | The preparation method of magnetic iron oxide/bagasse active carbon | |
| CN105347322B (en) | Spherical nano-porous hydroxylapatite prepared through shells and preparation method thereof | |
| CN107309438A (en) | A kind of method that polysaccharide from Phyllanthus Emblica L prepares Nano Silver compound particle | |
| CN110449127A (en) | A kind of charcoal base load iron composite material preparation method and applications | |
| CN105771900A (en) | Iron-loaded modified active carbon and preparation method thereof | |
| CN110142032A (en) | Chitosan biological carbon composite and its preparation method and application | |
| CN110369486A (en) | A kind of preparation and application of the sulfhydrylation charcoal of repairing heavy metal pollution water body and soil | |
| CN104056592B (en) | The preparation method of medical stone demanganization filtrate and the application for removing manganese ion in underground water thereof | |
| CN109092247A (en) | The preparation method of modified mulberry bar biomass carbon adsorbent | |
| CN101289380B (en) | Process for preparing calcium propionate by using japanese pearl oyster conch | |
| CN107522281A (en) | A kind of preparation method for the algicide administered for water-surface blue-green algae | |
| CN110589864A (en) | Method for preparing oyster nano calcium carbonate by taking trypsin as crystal form control agent | |
| CN109678696A (en) | A method of extracting lactic acid from hyaluronic acid fermentation liquor | |
| CN107416899B (en) | A kind of nano wire α-Bi2O3The preparation method of dusty material | |
| CN106219730B (en) | A kind of method of semen litchi recycling application | |
| CN107748153A (en) | A kind of penicillamine probe of fluorescent ultraviolet dual signal model and its application | |
| CN107868261A (en) | A kind of carboxymethyl chitosan/oxidized graphene composite aquogel and its preparation and application | |
| CN101856149A (en) | Method for improving quality of cigarette filter tip | |
| CN104193012B (en) | Silver nano-grain is utilized to promote the method that Phanerochaete chrysosporium removes heavy metal cadmium | |
| CN108212111B (en) | A kind of heavy metal-polluted soil absorption pulvis and its production method | |
| CN114940966B (en) | Preparation method and application of tomato root tip protoplast single-cell suspension |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191220 |
|
| RJ01 | Rejection of invention patent application after publication |