CN110511417B - Schisandra chinensis nanofiber composite preservative film and preparation method thereof - Google Patents
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
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- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/04—Alginic acid; Derivatives thereof
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- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
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Abstract
A schisandra chinensis nano-fiber composite preservative film and a preparation method thereof relate to a preservative film and a preparation method thereof. The invention aims to solve the problems that the existing preservative film can not effectively prevent foreign bacteria from polluting food and can not inhibit the proliferation of microorganisms. The schisandra chinensis nanofiber composite preservative film consists of a schisandra chinensis nanofiber membrane, a polyanion membrane and a polycation membrane. The preparation method comprises the following steps: (1) the surface of the schisandra chinensis nano cellulose membrane is self-assembled with a polyanion membrane and a membrane after anion treatment; (2) self-assembling a polycation membrane on the surface of the membrane after anion treatment to obtain a membrane after primary assembly; (3) repeating the steps from (1) to (2) for 2-3 times to obtain the schisandra chinensis nanofiber composite preservative film. The advantages are that: firstly, the antibacterial agent has extremely high antibacterial performance; the product can be used for meat preservation, and the preservation time at room temperature (20 deg.C) is prolonged to 5 days.
Description
Technical Field
The invention relates to a preservative film and a preparation method thereof.
Background
In recent years, with the continuous improvement of living standard of people, the problems of food safety and environmental protection become the focus of attention of people, and the fresh-keeping of the perishable fruits has higher requirements, so that the fresh-keeping effect is required to be achieved, and the fresh-keeping fruits are not damaged. Most of the existing packaging films in China are made of Polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyester and other materials, but because the packaging films contain carcinogenic chemical substances such as phthalate, di (2-ethyl) hexyl adipate (DEHA) and the like and cannot be degraded by the natural environment, the phenomena of environmental pollution, crop yield reduction, livestock death and the like are caused, and the adverse effects are caused on the life and the body health of people.
The existing preservative film only has a simple preservation function, the shelf life of food is prolonged by sealing and isolating air, but the pollution of external bacteria to preserved food cannot be effectively prevented, the proliferation of microorganisms cannot be inhibited, and the preservative film can only be used for preserving fruits and vegetables, and the preservation time at room temperature cannot exceed 2 days when being used for preserving meat.
Disclosure of Invention
The invention aims to solve the problems that the existing preservative film cannot effectively block the pollution of external bacteria to food and cannot inhibit the proliferation of microorganisms, and provides a schisandra chinensis nano-fiber composite preservative film and a preparation method thereof.
The schisandra chinensis nanofiber composite preservative film consists of a schisandra chinensis nano cellulose film, a polyanion film and a polycation film, wherein the polyanion film and the polycation film are sequentially self-assembled on the surface of the schisandra chinensis nano cellulose film.
The preparation method of the schisandra chinensis nanofiber composite preservative film is specifically completed according to the following steps:
(1) soaking the Chinese magnoliavine fruit nano cellulose membrane in polyanion solution with the concentration of 5 mg/mL-8 mg/mL, standing for 2 min-4 min at the temperature of 60-75 ℃, then taking out, cleaning and drying to obtain a membrane after anion treatment;
(2) soaking the film subjected to anion treatment in polycation solution with the concentration of 3-6 mg/mL, standing for 2-4 min at the temperature of 60-75 ℃, and then taking out, cleaning and drying to obtain the film after primary assembly;
(3) and repeating the film after the primary assembly for 2-3 times according to the steps six (1) to (2) to obtain the schisandra chinensis nanofiber composite preservative film.
The invention has the following principle and advantages: firstly, the roots and stems of schisandra chinensis are used as raw materials to prepare schisandra chinensis nanocellulose, and then the schisandra chinensis nanocellulose is used as raw materials to prepare the preservative film, the schisandra chinensis is a perennial Magnoliaceae plant, the roots and stems of the schisandra chinensis nanocellulose are rich in schisandra chinensis lignan, and the method disclosed by the invention keeps the schisandra chinensis lignan in the schisandra chinensis nanocellulose, so that the prepared preservative film is good in oxidation resistance and has extremely high antibacterial performance; secondly, the polyelectrolyte multilayer coating is prepared on the surface of the nano cellulose membrane in a self-assembly mode, so that the oxidation resistance and the bacteriostatic property of the preservative film are further improved; the schisandra chinensis nanofiber composite preservative film prepared by the invention can be used for meat preservation, pork is sealed by the schisandra chinensis nanofiber composite preservative film and preserved at room temperature (20 ℃), and the preservation time is prolonged to 5 days.
Detailed Description
The first embodiment is as follows: the embodiment is a schisandra chinensis nanofiber composite preservative film which consists of a schisandra chinensis nano cellulose film, a polyanion film and a polycation film, wherein the polyanion film and the polycation film are sequentially self-assembled on the surface of the schisandra chinensis nano cellulose film.
The second embodiment is as follows: the present embodiment differs from the first embodiment in that: the schisandra chinensis nano cellulose membrane is prepared by the following steps:
firstly, crushing: drying the Chinese magnoliavine fruit roots until the water content is lower than 10%, and then crushing until the particle size is 200-300 meshes to obtain Chinese magnoliavine fruit root and stem powder;
①, putting schisandra chinensis rhizome powder into a sodium hypochlorite solution with the concentration of 0.5-1 wt%, adjusting the pH to 4-5 by glacial acetic acid, heating and carrying out reflux reaction for 4-6 h to obtain a primary reaction product, ②, carrying out suction filtration on the primary reaction product, washing the primary reaction product by deionized water until the filtrate obtained by suction filtration is neutral to obtain a solid product, putting the solid product into a sodium sulfite solution with the concentration of 5-10 wt%, heating and carrying out reflux reaction for 4-6 h to obtain a secondary reaction product, ③, carrying out suction filtration on the secondary reaction product, washing the secondary reaction product by deionized water for 1-3 times to obtain a washed solid product, putting the washed solid product into a NaOH aqueous solution with the mass fraction of 5-30%, stirring and reacting for 30 min-90 min at the temperature of 40-100 ℃, and carrying out centrifugal separation to obtain a centrifugal solid, ④, adding the centrifugal solid into water under the assistance of ultrasound to obtain the schisandra chinensis nanocellulose suspension with the mass fraction of 2-3%;
thirdly, film preparation:
and (2) flowing 2-3% by mass of the schisandra chinensis nano cellulose suspension on a substrate, and drying at 40-50 ℃ to obtain the schisandra chinensis nano cellulose membrane. The rest is the same as the first embodiment.
The third embodiment is different from the second embodiment in that the ratio of the mass of the schisandra chinensis rhizome powder to the volume of the sodium hypochlorite solution with the concentration of 0.5 wt% -1 wt% in the second ① is 3g (10-30) mL, and the other steps are the same as the second embodiment.
Fourth embodiment the present embodiment is different from the second or third embodiment in that the ratio of the mass of the solid product to the volume of the sodium sulfite solution having a concentration of 5 to 10 wt% in the second ② is 1.5g (4 to 20) mL, and the other embodiments are the same as the second or third embodiment.
Fifth embodiment five the difference between this embodiment and any of the second to fourth embodiments is that the volume ratio of the mass of the washed solid product to the NaOH aqueous solution having a mass fraction of 5% to 30% in the second step ③ is 1g (10 to 30) mL.
The sixth specific implementation mode: the embodiment is a preparation method of a schisandra chinensis nanofiber composite preservative film, which is specifically completed according to the following steps:
(1) soaking the Chinese magnoliavine fruit nano cellulose membrane in polyanion solution with the concentration of 5 mg/mL-8 mg/mL, standing for 2 min-4 min at the temperature of 60-75 ℃, then taking out, cleaning and drying to obtain a membrane after anion treatment;
(2) soaking the film subjected to anion treatment in polycation solution with the concentration of 3-6 mg/mL, standing for 2-4 min at the temperature of 60-75 ℃, and then taking out, cleaning and drying to obtain the film after primary assembly;
(3) and repeating the film after the primary assembly for 2-3 times according to the steps from six (1) to (2) to obtain the schisandra chinensis nano cellulose film.
The seventh embodiment: the present embodiment differs from the sixth embodiment in that: the polyanion solution in the step (1) is one or a mixture of sodium polyacrylate and sodium alginate. Other embodiments are the same as the sixth embodiment
The specific implementation mode is eight: the difference between this embodiment and one of the sixth and seventh embodiments is: the polycation solution in the step (2) is chitosan. The rest is the same as the sixth embodiment or the seventh embodiment
The specific implementation method nine: the present embodiment differs from the sixth to eighth embodiments in that: the Chinese magnoliavine fruit nano cellulose membrane in the step (1) is prepared according to the following steps:
firstly, crushing: drying the Chinese magnoliavine fruit roots until the water content is lower than 10%, and then crushing until the particle size is 200-300 meshes to obtain Chinese magnoliavine fruit root and stem powder;
①, putting schisandra chinensis rhizome powder into a sodium hypochlorite solution with the concentration of 0.5-1 wt%, adjusting the pH to 4-5 by glacial acetic acid, heating and carrying out reflux reaction for 4-6 h to obtain a primary reaction product, ②, carrying out suction filtration on the primary reaction product, washing the primary reaction product by deionized water until the filtrate obtained by suction filtration is neutral to obtain a solid product, putting the solid yield into a sodium sulfite solution with the concentration of 5-10 wt%, heating and carrying out reflux reaction for 4-6 h to obtain a secondary reaction product, ③, carrying out suction filtration on the secondary reaction product, washing the secondary reaction product by deionized water for 1-3 times to obtain a washed solid product, putting the washed solid product into a NaOH aqueous solution with the mass fraction of 5-30%, stirring and reacting for 30 min-90 min at the temperature of 40-100 ℃, and carrying out centrifugal separation to obtain a centrifugal solid, ④, adding the centrifugal solid into water under the assistance of ultrasound to obtain the schisandra chinensis nanocellulose suspension with the mass fraction of 2-3%;
thirdly, film preparation:
and (2) flowing 2-3% by mass of the schisandra chinensis nano cellulose suspension on a substrate, and drying at 40-50 ℃ to obtain the schisandra chinensis nano cellulose membrane.
The rest is the same as the sixth to eighth embodiments
Tenth embodiment the difference between this embodiment and the ninth embodiment is that the ratio of the mass of the schisandra chinensis rhizome powder to the volume of the sodium hypochlorite solution having a concentration of 0.5 to 1 wt% in step two ① is 3g (10 to 30) ml.
The difference between this embodiment and either of the ninth and tenth embodiments is that the volume ratio of the mass of the solid product to the sodium sulfite solution having a concentration of 5 wt% to 10 wt% in step two ② is 1.5g (4 to 20) mL.
Twelfth embodiment the present embodiment is different from the ninth to eleventh embodiments in that the volume ratio of the mass of the washed solid product to the NaOH aqueous solution having a mass fraction of 5% to 30% in the second step ③ is 1g (10 to 30) mL, and the other embodiments are the same as the ninth to eleventh embodiments.
The following tests were carried out to confirm the effects of the present invention
Example 1: the preparation method of the schisandra chinensis nanofiber composite preservative film is specifically completed according to the following steps:
(1) soaking the schisandra chinensis nano cellulose membrane in polyanion solution with the concentration of 7mg/mL, standing for 4min at the temperature of 65 ℃, and then taking out, cleaning and drying to obtain a film after anion treatment;
(2) soaking the film subjected to anion treatment in a polycation solution with the concentration of 5mg/mL, standing for 4min at the temperature of 70 ℃, and then taking out, cleaning and drying to obtain a film subjected to primary assembly;
(3) repeating the film after the primary assembly for 2 times according to the steps from (1) to (2) to obtain the schisandra chinensis nanofiber composite preservative film;
the polyanion solution in the step (1) is sodium alginate;
the polycation solution in the step (2) is chitosan;
the Chinese magnoliavine fruit nano cellulose membrane in the step (1) is prepared according to the following steps:
firstly, crushing: drying the Chinese magnoliavine fruit roots until the water content is lower than 10%, and then crushing until the particle size is 200-300 meshes to obtain Chinese magnoliavine fruit root and stem powder;
①, putting schisandra chinensis rhizome powder into a sodium hypochlorite solution with the concentration of 1 wt%, adjusting the pH to 4 by adopting glacial acetic acid, heating and carrying out reflux reaction for 5h to obtain a primary reaction product, wherein the volume ratio of the mass of the schisandra chinensis rhizome powder to the sodium hypochlorite solution with the concentration of 1 wt% is 3g:14mL, ②, carrying out suction filtration on the primary reaction product, washing by using deionized water until the filtrate obtained by suction filtration is neutral to obtain a solid product, then putting the solid yield into a sodium sulfite solution with the concentration of 8 wt%, heating and carrying out reflux reaction for 6h to obtain a secondary reaction product, wherein the volume ratio of the mass of the solid product to the sodium sulfite solution with the concentration of 8 wt% is 1.5g:11mL, ③, carrying out suction filtration on the secondary reaction product, washing for 3 times by using the deionized water to obtain a washed solid product, putting the washed solid product into a NaOH aqueous solution with the mass fraction of 20%, stirring and carrying out reaction for 60min at the temperature of 80 ℃, carrying out centrifugal separation to obtain a solid product, and adding the washed aqueous solution of the nano cellulose into ④ g:22mL of the ultrasonic auxiliary aqueous solution with the mass of the NaOH;
thirdly, film preparation:
and (3) flowing the schisandra chinensis nano cellulose suspension on a substrate in percentage by mass, and drying at the temperature of 45 ℃ to obtain the schisandra chinensis nano cellulose film.
Comparative experiment: (1) soaking the glass substrate in a polyanion solution with the concentration of 7mg/mL, standing for 4min at the temperature of 65 ℃, then taking out, cleaning and drying to obtain a film after anion treatment;
(2) soaking the film subjected to anion treatment in a polycation solution with the concentration of 5mg/mL, standing for 4min at the temperature of 70 ℃, and then taking out, cleaning and drying to obtain a film subjected to primary assembly;
(3) and repeating the film after the primary assembly for 2 times according to the steps from six (1) to (2), and stripping the glass substrate to obtain the composite preservative film without the schisandra chinensis nanofibers.
The polyanion solution in the step (1) is sodium alginate.
The polycation solution in the step (2) is chitosan.
5 types of commercially available preservative films are purchased and respectively marked as a preservative film I, a preservative film II, a preservative film III, a preservative film IV and a preservative film V, and the preservative films are subjected to preservation detection with the schisandra chinensis nanofiber composite preservative film (marked as a composite preservative film) prepared in the example 1, the schisandra chinensis nanofiber membrane and the composite preservative film (marked as an anion and cation composite preservative film) which is prepared in a comparative experiment and does not contain schisandra chinensis nanofibers.
Selecting the pork fillet of a long white pig as a insurance object, and cutting the pork fillet into blocks to obtain pork blocks, wherein the thickness of the pork blocks is 0.8cm, the width of the pork blocks is 3cm, and the length of the pork blocks is 5 cm; the preservative film I, the preservative film II, the preservative film III, the preservative film IV, the preservative film V, the schisandra chinensis nano cellulose film, the schisandra chinensis nano fiber composite preservative film (marked as composite preservative film) prepared in the embodiment 1 and the composite preservative film (marked as anion and cation composite preservative film) which is prepared in a comparative experiment and does not contain schisandra chinensis nano fibers are sequentially selected for preservation at the preservation temperature of 20 ℃, the preservation is continuously carried out for 5 days, the preservation is divided into maturity, autolysis and decay according to the deterioration condition of the flesh quality, the preservation result is recorded, as shown in table 1, the table 1 shows that the schisandra chinensis nano fiber composite preservative film prepared in the embodiment can greatly prolong the maturation time and the autolysis time of the flesh quality at the temperature of 20 ℃, namely, the preservative film containing no schisandra chinensis nano fibers and the preservative film containing no schisandra chinensis nano fibers can prolong the preservation time, but the effect is not good compared with the schisandra chinensis nano-fiber composite preservative film.
TABLE 1
| 12h | 24h | 36h | 48h | 72h | 96h | 120h | |
| Preservative film I | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage | Spoilage | Spoilage |
| Preservative film II | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage | Spoilage | Spoilage |
| Preservative film III | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage | Spoilage | Spoilage |
| Preservative film IV | Maturation of the plant | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage | Spoilage |
| Preservative film V | Maturation of the plant | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage | Spoilage |
| Composite preservative film | Maturation of the plant | Maturation of the plant | Maturation of the plant | Maturation of the plant | Maturation of the plant | Autolysis | Autolysis |
| Anion and cation composite preservative film | Maturation of the plant | Maturation of the plant | Autolysis | Autolysis | Spoilage | Spoilage | Spoilage |
| Chinese magnoliavine fruit nano cellulose membrane | Maturation of the plant | Maturation of the plant | Maturation of the plant | Autolysis | Spoilage | Spoilage | Spoilage |
Staphylococcus aureus ATCC25923, Escherichia coli ATCC11229, Salmonella ATCC14028 and Proteus vulgaris CMCC (B)49027 are selected. 200 mu L of tested bacterium liquid (the bacterium content is 1 multiplied by 108CFU/mL) is transferred to a plate containing a culture medium in an aseptic environment, then a schisandra chinensis nanofiber composite preservative film with the diameter of 5mm, a schisandra chinensis nano cellulose film and a preservative film without schisandra chinensis nanofibers are placed in the middle of the plate, the plate is cultured for 24 hours in a constant-temperature incubator at the temperature of 37 ℃, and a bacteriostatic circle (mm) is tested, as shown in table 2, after the schisandra chinensis nanofibers, polyanions and polycations are synergistic, the bacteriostatic action is greatly improved.
TABLE 2
Claims (9)
1. The schisandra chinensis nanofiber composite preservative film is characterized by consisting of a schisandra chinensis nanofiber membrane, a polyanion membrane and a polycation membrane, wherein the polyanion membrane and the polycation membrane are sequentially self-assembled on the surface of the schisandra chinensis nanofiber membrane;
the schisandra chinensis nano cellulose membrane is prepared by the following steps:
firstly, crushing: drying the Chinese magnoliavine fruit roots until the water content is lower than 10%, and then crushing until the particle size is 200-300 meshes to obtain Chinese magnoliavine fruit root and stem powder;
①, putting schisandra chinensis rhizome powder into a sodium hypochlorite solution with the concentration of 0.5-1 wt%, adjusting the pH to 4-5 by glacial acetic acid, heating and carrying out reflux reaction for 4-6 h to obtain a primary reaction product, ②, carrying out suction filtration on the primary reaction product, washing the primary reaction product by deionized water until the filtrate obtained by suction filtration is neutral to obtain a solid product, putting the solid product into a sodium sulfite solution with the concentration of 5-10 wt%, heating and carrying out reflux reaction for 4-6 h to obtain a secondary reaction product, ③, carrying out suction filtration on the secondary reaction product, washing the secondary reaction product by deionized water for 1-3 times to obtain a washed solid product, putting the washed solid product into a NaOH aqueous solution with the mass fraction of 5-30%, stirring and reacting for 30 min-90 min at the temperature of 40-100 ℃, and carrying out centrifugal separation to obtain a centrifugal solid, ④, adding the centrifugal solid into water under the assistance of ultrasound to obtain the schisandra chinensis nanocellulose suspension with the mass fraction of 2-3%;
thirdly, film preparation:
and (2) flowing 2-3% by mass of the schisandra chinensis nano cellulose suspension on a substrate, and drying at 40-50 ℃ to obtain the schisandra chinensis nano cellulose membrane.
2. The schisandra chinensis nano-fiber composite preservative film according to claim 1, wherein the volume ratio of the mass of the schisandra chinensis rhizome powder to the sodium hypochlorite solution with the concentration of 0.5 wt% -1 wt% in step two ① is 3g (10-30) mL.
3. The schisandra chinensis nano-fiber composite preservative film according to claim 2, wherein the volume ratio of the mass of the solid product to the sodium sulfite solution with the concentration of 5 wt% -10 wt% in the second step ② is 1.5g (4-20) mL.
4. The schisandra chinensis nanofiber composite preservative film according to claim 1, 2 or 3, wherein the volume ratio of the mass of the washed solid product to the NaOH aqueous solution with the mass fraction of 5% -30% in step two ③ is 1g (10-30) mL.
5. The preparation method of the schisandra chinensis nano-fiber composite preservative film according to claim 1, which is characterized by comprising the following steps:
(1) soaking the Chinese magnoliavine fruit nano cellulose membrane in polyanion solution with the concentration of 5 mg/mL-8 mg/mL, standing for 2 min-4 min at the temperature of 60-75 ℃, then taking out, cleaning and drying to obtain a membrane after anion treatment;
(2) soaking the film subjected to anion treatment in polycation solution with the concentration of 3-6 mg/mL, standing for 2-4 min at the temperature of 60-75 ℃, and then taking out, cleaning and drying to obtain the film after primary assembly;
(3) and repeating the film after the primary assembly for 2-3 times according to the steps six (1) to (2) to obtain the schisandra chinensis nanofiber composite preservative film.
6. The preparation method of fructus Schisandrae chinensis nanometer fiber composite preservative film according to claim 5, characterized in that the polyanion solution in step (1) is one or a mixture of two of sodium polyacrylate and sodium alginate.
7. The preparation method of fructus Schisandrae chinensis nanometer fiber composite preservative film according to claim 5 or 6, characterized in that the polycation solution in step (2) is chitosan.
8. The preparation method of fructus schizandrae nanofiber composite preservative film according to claim 5, wherein the fructus schizandrae nanofiber membrane in the step (1) is prepared according to the following steps:
firstly, crushing: drying the Chinese magnoliavine fruit roots until the water content is lower than 10%, and then crushing until the particle size is 200-300 meshes to obtain Chinese magnoliavine fruit root and stem powder;
①, putting schisandra chinensis rhizome powder into a sodium hypochlorite solution with the concentration of 0.5-1 wt%, adjusting the pH to 4-5 by adopting glacial acetic acid, heating and carrying out reflux reaction for 4-6 h to obtain a primary reaction product, wherein the volume ratio of the mass of the schisandra chinensis rhizome powder to the sodium hypochlorite solution with the concentration of 0.5-1 wt% is 3g (10-30) mL, ②, carrying out suction filtration on the primary reaction product, washing with deionized water until filtrate obtained by suction filtration is neutral to obtain a solid product, putting the solid yield into a sodium sulfite solution with the concentration of 5-10 wt%, heating and carrying out reflux reaction for 4-6 h to obtain a secondary reaction product, wherein the volume ratio of the mass of the solid product to the solution with the concentration of 5-10 wt% is 1.5g (4-20) mL, ③, washing the secondary reaction product with deionized water for 1-3 times, obtaining a washed solid product, putting the washed solid product into an aqueous solution with the mass fraction of 5-10 wt% and carrying out centrifugal reaction on the aqueous solution of 5-30 min at the centrifugal reaction temperature of ④ -40 min to obtain a solid product, and centrifuging the aqueous solution of NaOH to obtain a solid product, and centrifuging the aqueous solution with the centrifugal reaction solution at the temperature of ④ -30 min and the centrifugal reaction to obtain the solid product of the solid product, and the centrifugal reaction product, wherein the centrifugal reaction product;
thirdly, film preparation:
and (2) flowing 2-3% by mass of the schisandra chinensis nano cellulose suspension on a substrate, and drying at 40-50 ℃ to obtain the schisandra chinensis nano cellulose membrane.
9. The preparation method of the schisandra chinensis nano-fiber composite preservative film according to claim 8, wherein the volume ratio of the mass of the washed solid product to the NaOH aqueous solution with the mass fraction of 5% -30% in step two ③ is 1g (10-30) mL.
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