Astaxanthin microcapsule and preparation method thereof

Abstract

The preparation method comprises the following steps: extruding an astaxanthin raw material medicine, copovidone VA64, glucose and other auxiliary materials in a specific proportion in a double-screw hot melt extruder to obtain an astaxanthin nano dispersion so as to enhance the water dispersibility and solubility of the astaxanthin nano dispersion; suspending the astaxanthin nano dispersion in a gelatin solution, and then sequentially carrying out high-speed homogenization and spray drying to obtain astaxanthin microcapsules; according to the preparation method disclosed by the invention, the solid astaxanthin is directly embedded by combining hot-melt extrusion and spray drying, so that the drug loading capacity of the microcapsule is greatly increased, and the solubility and the stability of the astaxanthin are enhanced.

Description

Technical Field

[0001] The invention relates to an astaxanthin microcapsule and a preparation method thereof. Background Art

[0002] Astaxanthin is a β-carotenoid that is mainly found in shrimp, crab and algae. Astaxanthin is a very strong antioxidant and a safe food additive. It can be used as a natural colorant for food and condiments. Because its structure contains a long conjugated double bond and unsaturated ketone and hydroxyl groups at the end of the conjugated double bond chain, it is easily affected by light, heat and oxygen and loses its activity. And because astaxanthin is a non-polar substance, insoluble in water and slightly soluble in oil, the development of astaxanthin preparations is somewhat difficult, which also greatly limits its application.

[0003] Microencapsulation technology is a technology that embeds solid, liquid, gas and other substances in capsule materials to form a tiny closed system. This method can not only isolate and protect the embedded substances from the influence of light and oxygen, slow down their oxidation and thus improve their stability, but also effectively cover up the special smell of the core material and control the release rate of the drug. Therefore, astaxanthin can be embedded through microencapsulation technology to improve its storage stability.

[0004] The preparation methods of microcapsules are divided into physical methods, chemical methods, and physical and chemical methods. The common ones include simple coagulation method, complex coagulation method, spray drying method, spray condensation method, suspension coating method, etc. Among them, spray drying technology is a widely used microcapsule preparation technology. This method disperses the liquid material into extremely fine mist particles through mechanical action, and then dries it by heating gas to finally obtain a dry particle product. The characteristics of spray drying technology are: the liquid material is directly converted into a powdered product, which can realize continuous batch production, and products with uniform particle size and good dispersion can be obtained without the need for a crushing process.

[0005] Since astaxanthin is relatively stable in oil solution, the current preparation methods of astaxanthin microcapsules basically adopt the method of dissolving astaxanthin in oil phase, emulsifying astaxanthin oil with water phase and then spray drying. For example, patent document CN111358762A uses astaxanthin oil as core material, and uses modified starch and β-cyclodextrin as capsule material for spray drying; CN101292965B dissolves astaxanthin in fatty acid glyceride substances, embeds it in gelatin for spray drying; CN1048056051 embeds astaxanthin oil with sodium methylcellulose and modified starch for spray drying, etc. However, this method has a low drug loading and a small content of effective ingredients, which makes the dosage of the preparation product large. Directly embedding astaxanthin solid into microcapsules is a method to increase the drug loading of microcapsules and reduce the dosage of products. However, the limitations of this method are: the stability of astaxanthin solid is poor, and the encapsulation rate requirement is high; the hydrophobicity of astaxanthin solid is too strong, and the suspension is difficult to suspend uniformly, resulting in greater difficulty in encapsulation and lower encapsulation rate; and the dispersed particle size of astaxanthin in the microcapsule is large, and the dissolution rate is slow, which affects its bioavailability. Therefore, it is necessary to use new technologies to enhance the water dispersibility of astaxanthin solid and reduce its particle size to reduce the difficulty of encapsulation.

[0006] Hot melt extrusion technology can nanosize drugs through the effects of shear force and heat, thereby improving the water dispersibility of drugs and improving their solubility. The present invention converts astaxanthin solid into nano-size and disperses it in a carrier through a preferred hot melt extrusion process, reduces its particle size, improves roundness and enhances its water dispersibility, reduces the difficulty of embedding, and then uses gelatin for embedding after fully dispersing it in water to obtain astaxanthin microcapsules with good solubility.

[0007] This technology solves the problem that astaxanthin solid is difficult to directly embed. The astaxanthin microcapsules with high drug loading are prepared by nano-processing astaxanthin solid, which have good stability and good water dispersibility and solubility. Summary of the invention

[0008] The present invention aims to provide an astaxanthin microcapsule and a preparation method thereof. The present invention obtains an astaxanthin microcapsule with good stability and water dispersibility by combining hot melt extrusion technology with spray drying technology.

[0009] The technical solution of the present invention is as follows:

[0010] An astaxanthin microcapsule is prepared from the following raw materials in percentage by mass: 5-25% astaxanthin, 1.67-8.33% PVP VA64, 3.33-16.67% glucose, and 50-90% gelatin; the total amount of all raw materials is 100%.

[0011] Furthermore, the mass ratio of astaxanthin, PVP VA64 and glucose is preferably 3:1:2.

[0012] The preparation method of the astaxanthin microcapsules of the present invention is as follows:

[0013] (1) uniformly mixing astaxanthin, PVP VA64 and glucose, extruding the mixture in a twin-screw hot melt extruder, and dispersing the obtained extrudate in water after cooling to room temperature to obtain an extrudate suspension;

[0014] The process conditions of hot melt extrusion were: extrusion temperature 100 °C, screw speed 30 rpm;

[0015] The preferred mass ratio of extrudate to water is 1:20;

[0016] (2) adding gelatin into water to swell and dissolve it, thereby obtaining a gelatin solution;

[0017] The preferred mass ratio of gelatin to water is 1 to 5:20;

[0018] (3) adding the gelatin solution obtained in step (2) and the extrudate suspension obtained in step (1) into a mortar, mixing and grinding, then high-speed homogenizing, and finally spray drying to obtain astaxanthin microcapsules;

[0019] The preferred mixing and grinding time is 5 min;

[0020] The preferred high-speed homogenization parameter settings are: 10000 rpm, 3 to 15 min;

[0021] The spray drying parameters are preferably set as follows: drying temperature 130-190° C., feed rate 1.6-6.4 mL / min, and atomizing gas flow rate 439 L / h.

[0022] The technical points of the present invention are:

[0023] 1. In the raw material formula, the ratio of astaxanthin, PVP VA64 and glucose must be 3:1:2, otherwise nano-scale dispersion cannot be formed, and the dispersibility and solubility cannot be effectively improved. This is because PVP VA64 plays the role of separating and isolating the drug (astaxanthin) during the hot melt extrusion process; glucose acts as a filler and increases the torque of the extruder. The drug is sheared and broken into nano-scale during the extrusion process. The appropriate proportion of glucose can increase the extruder torque and increase the shear force, while PVP VA64 effectively isolates the broken drug and keeps the particle size of the drug at the nano-scale.

[0024] 2. The extrusion temperature should be 100°C. Too low a temperature will result in excessive torque, causing the extruder to stop, while too high a temperature will result in too small a torque and insufficient shear force, making it impossible to achieve nano-scale dispersion.

[0025] The beneficial effects of the present invention are:

[0026] The present invention realizes direct embedding of solid astaxanthin by combining hot-melt extrusion and spray drying, greatly increases the drug loading of microcapsules, and enhances the solubility and stability of astaxanthin. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 : Water dispersion of the drug substance (a) and microcapsules (b).

[0028] Figure 2 : Microcapsule water dispersion particle size data; (a) - intensity average data, (b) - number average data, (c) - volume average data.

[0029] Figure 3 : Results of UV stability tests of API and microcapsules. DETAILED DESCRIPTION

[0030] The present invention is further described below by means of specific embodiments, but the protection scope of the present invention is not limited thereto.

[0031] In the following embodiments,

[0032] Astaxanthin API comes from Zhejiang Pharmaceutical, and PVP VA64 uses BASF VA64.

[0033] Embodiment 1:

[0034] The astaxanthin raw material was mixed with PVP VA64 and glucose at a mass ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0035] 3 g of gelatin was added to 20 ml of water and the mixture was swelled and dissolved in a 37°C water bath. 1.04 g of the extrudate was added to 20 ml of water and the mixture was dispersed in a 37°C water bath.

[0036] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 10 minutes.

[0037] The homogenized suspension was spray dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0038] Embodiment 2:

[0039] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0040] 4 g of gelatin was added to 20 ml of water to swell and dissolve, and 1.04 g of the extrudate was added to 20 ml of water to disperse.

[0041] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0042] The homogenized suspension was spray dried at an inlet air temperature of 130° C., a feed rate of 1.6 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0043] Embodiment 3:

[0044] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0045] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0046] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0047] The homogenized suspension was spray dried at an inlet air temperature of 130° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0048] Embodiment 4:

[0049] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0050] 4 g of gelatin was added to 20 ml of water and the mixture was swelled and dissolved in a 37°C water bath. 1.04 g of the extrudate was added to 20 ml of water and the mixture was dispersed in a 37°C water bath.

[0051] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 3 minutes.

[0052] The homogenized suspension was spray dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0053] Embodiment 5:

[0054] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0055] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0056] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0057] The homogenized suspension was spray dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0058] Embodiment 6:

[0059] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0060] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0061] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 15 minutes.

[0062] The homogenized suspension was spray-dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0063] Embodiment 7:

[0064] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0065] 5 g of gelatin was added to 20 ml of water and the mixture was swelled and dissolved in a 37°C water bath. 1.04 g of the extrudate was added to 20 ml of water and the mixture was dispersed in a 37°C water bath.

[0066] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 10 minutes.

[0067] The homogenized suspension was spray dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0068] Embodiment 8:

[0069] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0070] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0071] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0072] The homogenized suspension was spray-dried at an inlet air temperature of 170° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0073] Embodiment 9:

[0074] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0075] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0076] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0077] The homogenized suspension was spray dried at an inlet air temperature of 190° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0078] Embodiment 10:

[0079] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0080] 4 g of gelatin was added to 20 ml of water and the mixture was swelled and dissolved in a 37°C water bath. 1.04 g of the extrudate was added to 20 ml of water and the mixture was dispersed in a 37°C water bath.

[0081] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 10 minutes.

[0082] The homogenized suspension was spray dried at an inlet air temperature of 150° C., a feed rate of 3.2 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0083] Embodiment 11:

[0084] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0085] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0086] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0087] The homogenized suspension was spray dried at an inlet air temperature of 130° C., a feed rate of 4.8 L / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0088] Embodiment 12:

[0089] The astaxanthin raw material was mixed with PVP VA64 and glucose at a ratio of 3:1:2, and the mixture was extruded in a twin-screw hot melt extruder at a screw speed of 30 rpm and an extrusion temperature of 100°C.

[0090] 4 g of gelatin was added to 20 ml of water for swelling and dissolution, and 1.04 g of the extrudate was added to 20 ml of water for dispersion.

[0091] The gelatin solution and the extrudate suspension were added to a mortar and mixed and ground for 5 minutes, and the ground suspension was homogenized at 10,000 rpm for 5 minutes.

[0092] The homogenized suspension was spray dried at an inlet air temperature of 130° C., a feed rate of 6.4 ml / min, and an atomizing gas flow rate of 439 L / h, and the finished product was collected.

[0093] Embodiment 13:

[0094] Investigation of drug loading and encapsulation efficiency: The microcapsules prepared in Examples 1-12 were dissolved in dimethyl sulfoxide, and their absorbance was measured at a wavelength of 481 nm to calculate their content; the microcapsules were washed with dichloromethane, and the amount of astaxanthin in the washing liquid was measured as unencapsulated astaxanthin. The drug loading and encapsulation efficiency were calculated, and the results are shown in Table 1.

[0095] Table 1: Drug loading and encapsulation efficiency of microcapsules in Examples 1-12

[0096] Example Drug loading Encapsulation efficiency Example 1 13.10％ 48.46％ Example 2 11.04％ 61.80％ Example 3 10.27％ 53.20％ Example 4 11.77％ 59.43％ Example 5 11.26％ 68.58％ Example 6 10.99％ 68.91％ Example 7 9.40％ 51.17％ Example 8 10.54％ 57.30％ Example 9 10.64％ 57.80％ Example 10 10.98％ 69.04％ Embodiment 11 10.85％ 68.89％ Example 12 11.68％ 70.03％

[0097] Water dispersibility and solubility investigation: Astaxanthin API and microcapsules of Example 12 were dispersed in water, and the dispersion state was observed by photographing; the microcapsules were filtered using a 0.22 um filter membrane, and the absorbance was measured at a wavelength of 481 nm to calculate the solubility. Figure 1 As shown in Table 2, astaxanthin raw materials have strong hydrophobicity. After being added to water, they all float on the water surface and cannot enter the water. The solubility in water is measured to be only 0.006μg / mL. After astaxanthin microcapsules are added to water, they can be quickly dispersed in water, and the entire water body is red. Its solubility is measured to be 28μg / mL.

[0098] Table 2: Solubility of API and microcapsules

[0099] Microcapsules Solubility: 28ug / ml API Solubility: 0.006ug / ml

[0100] Astaxanthin dispersion particle size determination: Astaxanthin microcapsules were dispersed in water and the particle size was measured using a Malvern particle size analyzer. Figure 2 It can be seen that the intensity average, volume average and number average all reach the nanometer level, which further proves that its water dispersibility is enhanced by extrusion technology.

[0101] Table 3: Particle size of microcapsules dispersed in water

[0102] Strong Average Average Particle size 449.6nm 286.5nm 798.4nm

[0103] Stability test: The API and the microcapsules of Example 12 were placed under direct ultraviolet light for 5 days, and their retention rates were measured every day. The results are shown in Table 1. Figure 3 .

[0104] Comparative Example 1:

[0105] 3 g of gelatin and 1 g of glucose were added into 20 ml of water for swelling and dissolution; 0.5 g of astaxanthin solid was added into 20 ml of water for dispersion.

[0106] The astaxanthin dispersion was added to the gelatin glucose solution and mixed evenly, and the suspension was homogenized at 10,000 rpm for 5 min.

[0107] The homogenized suspension was spray-dried at an inlet air temperature of 130° C., a feed rate of 3.2 ml / min, and an atomizer flow rate of 439 L / h, and the finished product was collected.

[0108] Comparative Example 2:

[0109] 3 g gelatin and 1 g glucose were added into 20 ml water for swelling and dissolution; 0.5 g astaxanthin solid and 0.02 g Tween 20 were added into 20 ml water for dispersion.

[0110] The dispersion of astaxanthin and Tween 20 was added to the gelatin glucose solution and mixed evenly, and the suspension was homogenized at 10000 rpm for 5 min.

[0111] The homogenized suspension was spray-dried at an inlet air temperature of 130° C., a feed rate of 3.2 ml / min, and an atomizer flow rate of 439 L / h, and the finished product was collected.

[0112] Comparative Example 3:

[0113] Add 3g gelatin and 1g glucose into 20ml water to swell and dissolve.

[0114] The astaxanthin solid was directly added to the gelatin glucose solution and ground for 5 min, and the suspension was homogenized at 10000 rpm for 5 min.

[0115] The homogenized suspension was spray-dried at an inlet air temperature of 130° C., a feed rate of 3.2 ml / min, and an atomizer flow rate of 439 L / h, and the finished product was collected.

[0116] Comparative Example 4:

[0117] 3 g of gelatin and 1 g of glucose were added into 20 ml of water for swelling and dissolution; and astaxanthin was pre-grinded for 5 min.

[0118] The gelatin glucose solution was added to the ground astaxanthin and mixed and ground for 5 minutes, and the suspension was homogenized at 10000 rpm for 5 minutes.

[0119] The homogenized suspension was spray-dried at an inlet air temperature of 130° C., a feed rate of 3.2 ml / min, and an atomizer flow rate of 439 L / h, and the finished product was collected.

[0120] The microcapsules in Comparative Examples 1-4 were dissolved with dimethyl sulfoxide, and their absorbance was measured at a wavelength of 481 nm to calculate their content; the microcapsules were washed with dichloromethane, and the amount of astaxanthin in the washing solution was measured as unencapsulated astaxanthin. The drug loading and encapsulation efficiency were calculated and the results are shown in Table 4.

[0121] Table 4: Drug loading and encapsulation efficiency of microcapsules in comparative examples 1-4

[0122] Comparative Example Drug loading Encapsulation efficiency Comparative Example 1 10.72％ 23.98％ Comparative Example 2 10.25％ 30.60％ Comparative Example 3 8.50％ 30.74％ Comparative Example 4 10.54％ 51.85％

[0123] It was found that compared with the example, the encapsulation rate was significantly lower than that of the example, indicating that the hot melt extrusion technology effectively reduced the difficulty of encapsulating astaxanthin solids and improved the encapsulation rate of the microcapsules.

Claims

1. An astaxanthin microcapsule, characterized in that: The invention is prepared from the following raw materials in percentage by mass: 5-25% astaxanthin, 1.67-8.33% PVPVA64, 3.33-16.67% glucose and 50-90% gelatin; the total percentage of all raw materials is 100%.

2. The astaxanthin microcapsule according to claim 1, characterized in that The mass ratio of astaxanthin, PVP VA64 and glucose is 3:1:

2.

3. The method for preparing astaxanthin microcapsules according to claim 1, characterized in that: The preparation method is as follows: (1) uniformly mixing astaxanthin, PVP VA64 and glucose, extruding the mixture in a twin-screw hot melt extruder, and dispersing the obtained extrudate in water after cooling to room temperature to obtain an extrudate suspension; (2) adding gelatin into water to swell and dissolve it to obtain a gelatin solution; (3) adding the gelatin solution obtained in step (2) and the extrudate suspension obtained in step (1) into a mortar, mixing and grinding, then high-speed homogenizing, and finally spray drying to obtain astaxanthin microcapsules.

4. The preparation method according to claim 3, characterized in that: In step (1), the process conditions of hot melt extrusion are: extrusion temperature 100° C., screw speed 30 rpm.

5. The preparation method according to claim 3, characterized in that: In step (1), the mass ratio of the extrudate to water is 1:

20.

6. The preparation method according to claim 3, characterized in that: In step (2), the mass ratio of gelatin to water is 1 to 5:

20.

7. The preparation method according to claim 3, characterized in that: In step (3), the mixing and grinding time is 5 minutes.

8. The preparation method according to claim 3, characterized in that: In step (3), the parameters of high-speed homogenization are set as: 10000 rpm, 3 to 15 min.

9. The preparation method according to claim 3, characterized in that: In step (3), the spray drying parameters are set as: drying temperature 130-190° C., feed rate 1.6-6.4 mL / min, and atomizing gas flow rate 439 L / h.

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