KR20200027346A - stone cell isolation method with high efficiency - Google Patents

Abstract

The present invention relates to a method for separating stone cells with increased efficiency. The method for separating stone cells according to the present invention does not use a chemical component, thereby having high safety. Also, the method has an excellent separating amount of stone cells and requires less time for separating the stone cells. Accordingly, the method can be usefully used for various processes requiring the stone cells.

Description

{Stone cell isolation method with high efficiency}

The present invention relates to a method for separating stone cells with increased efficiency.

Micro plastic refers to a small plastic of less than 5 mm and is included in toothpaste, detergent, and scrub, which are easily accessible in everyday life, and is known to contain approximately 2.8 million micro plastics in a 150 ml product.

However, the micro plastic is too small to be filtered by the sewage treatment facility, and flows into the sea and river. For example, according to a paper published in Science magazine in 2015, the amount of plastic waste introduced into the sea in 2010 is approximately 4.8 million to 12.7 million tons. Since this plastic is a petroleum compound containing polyethylene (PE), polypropylene (PP), and nylon, it encounters contaminants and causes new environmental problems, and the discarded plastic may turn into micro plastic over time. According to a paper published in the UK in 2015, the International List on Small Plastic Waste in the Ocean, it is estimated that there are at least 15 to 51 trillion micro plastics in the sea.

Micro plastic not only destroys the environment, but also poses a problem in that it threatens human health. This is because micro plastics are small in size, making it difficult to distinguish them from plankton, which is a food for fish or sea algae. Micro plastic is recognized as a serious problem because it causes bowel obstruction and can adversely affect energy allocation reduction and growth.

As a result of efforts by many researchers to find natural or biodegradable materials that can replace micro plastics, which are environmentally harmful substances, it was confirmed that seok cells could replace such abrasive materials.

The calcite cells, called reinforcing cells or posterior wall cells, have shorter, thicker, more irregular secondary cell walls that are shorter than fibrous cells. Stone cells are found in fruit tissue, especially in the flesh of the genus Pyrus. Stone cells are a type of lignification in which lignin is deposited on cell walls in tissues, and are known to act as a framework for firmly strengthening the cell walls so as not to change the shape of the cell walls. The main components of pear fruit endothelial cells were studied by Ranadive and Harard in Western pears a long time ago. The main components are lignocellulose, and the content of each component is roughly carbohydrates (cellulose, hemicellulose), like the primary cell wall. , Pectin) 82% and contains 18-35% lignin that is not in the primary cell wall. In addition, the main units of lignin are vanillin and syringaldehyde, and in the case of carbohydrates, glucose and xylose are called.

On the other hand, pear processing is mainly pear production, and clear pear juice (heated at 80-90 ° C for 30 minutes-2 hours) and heated pear juice (high pressure heating at 120 ° C for 2-4 hours) are produced by two methods. Baebak, which is produced in the production of pear juice, accounts for 20-35%, but most of it is recycled as compost, and there are no processed products using it. In addition, pears have become a problem due to a large number of fallen fruits and non-products due to millet disasters during the first half of the growth period, such as typhoons.

Thus, the inventors of the present invention, as a result of diligent research efforts to develop a method for efficiently obtaining a stone cell that can replace microplastics, show that the method of separating the petroleum cells according to the present invention shows a high amount of separating cells and low requirements. The present invention was completed by confirming that it had time.

One object of the present invention is 1) washing the pear (residual residue after pear juice extraction); 2) grinding the washed pear; 3) dehydrating and drying the ground pear; 4) pulverizing the dry pear to powder; And 5) to provide a method for separating stone cells comprising the step of separating the stone cells and the pulp residue.

Another object of the present invention is to provide a stone cell obtained by the above method.

One aspect of the present invention for achieving the above object is, 1) washing the pear (residual residue after pear extract); 2) grinding the washed pear; 3) dehydrating and drying the ground pear; 4) pulverizing the dry pear to powder; And 5) provides a method for separating stone cells comprising the step of separating the stone cells and the pulp residue.

In the present invention, the term, "baking" refers to a by-product left after juice from a pear, and there is a raw pear remaining as a by-product remaining after juice and a raw pear remaining after juice by a heat pressing method, but is not limited thereto.

The bakbak may be used for peeling, peeling or pulp of a pear, and may be specifically peeling or pulp. In addition, the pear that is the material of the pear bak can be used without limitation, such as harvested, cultivated or commercially available.

The term "stone cell" in the present invention is called a reinforcing cell or a posterior wall cell, and has a very hard secondary cell wall that is shorter, thicker and irregular than a fibrous cell. Stone cells are found in fruit tissue, especially in the flesh of the genus Pyrus. Stone cells are a type of lignification in which lignin is deposited on cell walls in tissues, and are known to act as a framework for firmly strengthening the cell walls so as not to change the shape of the cell walls. The main components of pear fruit endothelial cells were studied by Ranadive and Harard in Western pears a long time ago. The main components are lignocellulose, and the content of each component is roughly carbohydrates (cellulose, hemicellulose), like the primary cell wall. , Pectin) 82% and contains 18-35% lignin that is not in the primary cell wall. In addition, the main units of lignin are vanillin and syringaldehyde, and in the case of carbohydrates, glucose and xylose are called.

The washing step of 1) may be to remove impurities such as debris, fruit powder, calyx, and the like remaining in the pear by using water. It is difficult to remove foreign substances, such as seeds or debris, in the subsequent process, so the process of removing foreign substances in the washing process is an important step in determining the purity of seok cells.

In addition, the washing step may be to remove the sugar contained in the pear. When drying without a washing process in a situation where the sugar concentration of the bakbak is high, the hardness of the bakbak may be increased, and thus the grinding process required for the petroleum cell extraction process may be very difficult. The sugar removal process is a process required for the extraction of stony cells because the production of fine and uniform results in the grinding process determines the efficiency of stony cell extraction.

The washing step may be to use brine or distilled water.

Specifically, the washing step may be to use 10 to 30% salt water.

When salt water is used in the washing step, the separation efficiency of seok cells can be increased through primary filtering of suspended solids and sediments using a concentration gradient.

The amount of water used in the washing step may be 2 to 8 times, specifically 3 to 6 times, and more specifically 3 to 5 times, as a volume ratio, but is not limited thereto.

The grinding step of 2) may be to increase the separation efficiency of seok cells through the process of finely separating the pear.

The dehydration step of 3) may be dehydration using centrifugation.

The dehydration step may be used for centrifugation for the purpose of lowering the moisture content in the bakbae in order to increase the drying efficiency of the stone cell separation process, the centrifugal force required for dehydration is 500 to 15000rpm, specifically 600 to 10000rpm, more specifically 700 to 5000rpm , May be more specifically 800 to 3000 rpm, the dehydration time is 1 minute to 60 minutes, specifically 2 minutes to 40 minutes, more specifically 3 minutes to 20 minutes, and more specifically 5 minutes to 10 minutes, but limited thereto Does not work.

The drying step of 3) may be hot air drying or natural drying.

The temperature of the hot air drying may be 50 ° C to 80 ° C, specifically 55 ° C to 75 ° C, more specifically 60 ° C to 70 ° C, but is not limited thereto.

In addition, when drying in hot air, it is possible to easily spread the pear to dry.

The grinding step of 4) can completely remove the flesh attached to the stony cells in the process of grinding and pulverizing the completely dried pear.

In the separation step 5), the pulverized stone cells and the pulp residue may be separated using a vibrating body. In addition, the separating step may further include filtering the stone cells by size using the perforation size of the vibrating body.

Another aspect of the present invention provides stone cells isolated by the above method.

In a specific embodiment of the present invention, as a result of attempting and analyzing various separation methods, seok cells were obtained through the most efficient seok cell separation method.

This suggests that it is possible to provide stone cells obtained through the above-described separation method.

The method for separating seok cells according to the present invention does not use a chemical component, and thus has high safety, has an excellent amount of separating cells, and has a low time for separating seok cells, and thus can be effectively used in various processes requiring seok cells.

1 is a view showing the shape of a stone cell according to the separation method.
2 is a diagram schematically illustrating a process for extracting a stone cell.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1. Preparation of samples

Baebak (residue left after extract of pear juice), ripe raw pear, can be used, but bakbak made under high temperature conditions is easy to separate seok cells, so bakbak was used to separate seok cells.

Example 2. Exploration of optimal stony cell separation conditions

In order to confirm the optimal conditions for spheroid cell separation, spheroid cells were separated through various methods and the efficiency was compared.

2-1. Separation of stone cells by chemical method

A chemical method of dissolving substances such as dietary fibers around the stony cells using chemicals NaCl, NaOH, and HCl was used for pear fruit juice. Specifically, the pear was dissolved with 0.1 M NaCl while homogenizing for 10 minutes, washed with distilled water, added with 0.1 N NaOH, stirred for 1 minute, and washed with distilled water. Again, 0.5 N HCl was added and stirred, followed by washing with distilled water and repeating this process 2-3 times, followed by drying the remaining material to obtain seok cells. At this time, the chemical solution and the batch were mixed in a ratio of 1: 1.

2-2. Separation of stone cells using physical methods

The physical method was attempted by three methods constructed by applying various methods such as drying and grinding. The first method was to remove sugar by immersion in water after crushing, and then sieved and dried. The second method was dried, crushed, immersed in water to remove sugar, sieved and dried. The third method is a method using a centrifuge, first homogenized in 50% methanol for 30 minutes, then centrifuged at 12,000 rpm for 4 minutes, discarded the supernatant and dried.

2-3. Separation of stone cells using biological methods

A method of separating seok cells using enzymes such as cellulase and pectinase was used. Specifically, the pear was crushed in a blender, immersed to remove sugar, and then filtered through a sieve to add cellulase and pectinase at a concentration of 0.1%, treated for 24 hours, and then filtered through a sieve and dried.

The above methods are summarized in Table 1.

Chemical method Physical method Biological method Method 1 Method 2 Method 3 Homogenization of 0.1 M NaCl, 10 min Grinding (mixer) dry Homogenization of 50% methanol (1: 1), 30 minutes Juice ↓ ↓ ↓ ↓ ↓ 0.1 N NaOH 10 min Sugar removal (immersion) Grinding (mixer) Centrifugation, 12,000 rpm, 4 minutes Grinding (mixer) ↕Repeat 3 ↓ ↓ ↓ ↓ 0.1 N HCl 10 min Sift Sugar removal (immersion) Remove supernatant Sugar removal (immersion) ↓ ↓ ↓ ↓ ↓ Washing distilled water 3 times dry Sift dry Enzyme treatment ↓ ↓ ↓ dry dry Sift ↓ dry

In addition, the content of seok cells obtained through the above method, the time required and the content by size are summarized in Tables 2 to 4.

Way Weight (g / 100g) Chemical method 7.14 ± 0.26 Physical method Method 1 11.5 ± 0.93 Method 2 16.9 ± 1.95 Method 3 25.7 ± 3.51 Biological method 19.3 ± 1.87

As shown in Table 2, the content of the isolated seok cells showed the lowest value in the chemical method, and the physical method 3 using the centrifugation showed the highest content, but the method using the centrifugation showed that the seok cells were clean. It was not separated.

Way Time required (minutes) Chemical method 188 ± 2.52 Physical method Method 1 119 ± 5.13 Method 2 739 ± 1.15 Method 3 78 ± 7.63 Biological method 1547 ± 15.3

In addition, as shown in Table 3, the time required for the separation of seok cells took the least time for the physical method 3 using centrifugation, and the longest time for the biological method.

Way Size (mm)
(ratio (%)) 0.5-0.25 0.25-0.1 0.1 or less Chemical method 0.52 ± 0.03
(7.3 ± 0.40) 5.29 ± 0.21
(74.1 ± 0.36) 1.33 ± 0.07
(18.6 ± 0.73) physical
Way

Method 1 2.46 ± 0.17
(21.5 ± 2.74) 8.78 ± 0.95
(76.6 ± 2.35) 0.23 ± 0.06
(2.0 ± 0.39) Method 2 1.88 ± 0.47
(11.2 ± 3.77) 14.4 ± 1.96
(85.7 ± 2.06) 0.53 ± 0.34
(3.1 ± 1.71) Method 3 5.68 ± 0.23
(22.1 ± 3.81) 14.2 ± 2.70
(55.4 ± 3.08) 5.76 ± 1.02
(22.4 ± 0.88) Biological method 4.81 ± 0.29
(24.9 ± 3.32) 11.0 ± 1.71
(56.8 ± 4.97) 3.54 ± 0.67
(18.3 ± 2.69)

As shown in Table 4, the physical method 3 using a centrifuge or the biological method using an enzyme confirmed that the content of seok cells below 0.5-0.25mm or 0.1mm was relatively high, and seok cells of approximately 0.25-0.1mm were generally used. It was confirmed that the content was high.

Finally, in order to confirm that the removal of the flesh from the stalk cells was good, the separated stalk cells were photographed and compared.

As shown in FIG. 1, it can be confirmed that the biological method was most cleanly separated, and the physical method 3 using a centrifuge was confirmed to be unclean.

 Therefore, the chemical method has the disadvantage that the amount of separation is small, and the biological method has the disadvantage that the time required for the separation of seok cells is large, so it was determined that the method of separating seok cells through the physical method is the most efficient. However, unlike physical methods 1 and 2, in the case of 3, the amount of separation is large and the time required is short, but there is a disadvantage that the flesh is not separated cleanly. To compensate for this, physical methods 1 to 3 are combined to derive the final extraction process as shown in FIG. 2. .

As a result of separating seok cells by the extraction process of Figure 2, as shown in Table 5, while having a segregation amount of seok cells similar to physical method 2, the time required for separation is 74 ± 12.7 minutes, which has a short time for physical method 3 , It was confirmed that it is the most efficient method because it is possible to cleanly separate seok cells.

Way Weight (g / 100g) Time required (minutes) Final extraction method 16.9 ± 1.95 74 ± 12.7

In the present specification, those skilled in the art of the present disclosure can fully recognize and infer the details thereof, and the detailed descriptions thereof have been omitted, and the technical spirit or essential configuration of the present invention may be changed in addition to the specific examples described herein. More variations are possible without departing from the scope of the invention. Therefore, the present invention can be implemented in a manner different from that specifically described and illustrated herein, which can be understood by those skilled in the art.

Claims (5)

1) washing pear (residual residue after pear juice extraction); 2) grinding the washed pear; 3) dehydrating and drying the ground pear; 4) pulverizing the dry pear to powder; And 5) separating the stone cells from the pulp residue.
According to claim 1,
The washing step of 1) is to use 10 to 30% of salt water, the method of separating seok cells.
According to claim 1,
Dehydration step of the 3) is to dehydrate by centrifugation for 1 minute to 60 minutes at 500 to 15000rpm, stony cell separation method.
According to claim 1,
Separation step of 5) is to further include the step of filtering the size of the stone cells, stone cell separation method.
Stone cells obtained by the method of any one of claims 1 to 4.

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