KR101722829B1 - Method of Removing Residual Agricultural Chemicals from Plant - Google Patents

Abstract

The present invention relates to the removal of pesticide residues in plants, and more particularly, since the crop protection agent component is effectively removed and the yield is maintained by thin-film distillation after supercritical extraction compared to the case where the above step is not applied, The resulting extract can be used in various fields such as a pharmaceutical composition, a cosmetic composition, a health functional food or an animal feed, or the ginseng fruit or the ginseng leaf oil.

Description

TECHNICAL FIELD The present invention relates to a method for removing residual agricultural chemicals from plants,

The present invention relates to a method for removing residual pesticides in plants, and more particularly, to a method for effectively removing residual pesticides by applying supercritical extraction and thin-film distillation.

Ginseng contains various kinds of ginsenosides. These ginsenosides are paraxadiol, pyrazyltriol and oleic acid. In addition to the above ginsenoside components, ginseng includes carbohydrates such as starch, antioxidative aromatic compounds of polyacetylenes, hypericin that protects the liver, and acid peptides that act like insulin. The main pharmacological actions of ginsenoside affect the central nervous system, the endocrine system, the immune system, the metabolic system, etc., and exert various effects on the body regulating function.

Ginseng has roots and primordia under the ground, and the ground consists of stem, leaf and fruit. Ginseng begins to bloom in ginseng for more than 3 years, and in June it produces a greenish-colored fruit. In the middle of July, the fruit of the ginseng will be red and be fruitless. Usually, ginseng is cultivated for more than 4 years, and the highest quality ginseng is known to have grown for 6 years. As such, it takes a long time to cultivate, and during the cultivation, due to various reasons such as various diseases and natural disasters, the yield is lower than that of the first cultivation, the price is high, and it takes much time to obtain ginseng. Therefore, it has been suggested to utilize the byproducts generated in the cultivation of ginseng.

For example, in the ginseng cultivation farm, when the ginseng fruit starts to be produced, the ginseng fruit is harvested to promote the growth of the ginseng root, and since the harvested fruit contains the ginsenoside component, the fertilizer of the ginseng cultivation field, . However, only a part is used, and most of them are recognized as garbage and discarded. In addition, since the ginseng fruit contains a crop protection agent component, there is a problem that it is not easy to use. On the other hand, ginseng leaves contain saponin similar to saponin of ginseng and contain 5 ~ 7 times of saponin which has pharmacological effect than ginseng. Therefore, it is evaluated as food or medicinal resource, but it is difficult to use because it contains ingredients of crop protection agent, It is.

As described above, although the ginseng fruit and ginseng leaf, which are present in the upper part of the present ginseng, contain ginsenoside components such as ginseng root, they are mostly discarded after being removed from ginseng, and the crop protecting agent components contained in ginseng and ginseng leaves It is difficult to remove ginseng leaves and the use of ginsenosides contained in ginseng leaves and ginseng leaves is insufficient.

Patent Document 1 discloses that ginseng leaves are treated by mashing to increase the content of ginsenoside, and in Patent Document 2, the ginseng fruit is matured and dried, And increasing the ginsenoside content by using an enzyme. Patent Document 3 discloses a method for removing a crop protection agent from a ginkgo extract. Patent Document 4 discloses a method for removing a crop protection agent from a ginkgo extract by extracting a plant with a polar solvent, Removal / purification. However, research has not been conducted on removing crop protection agents from ginseng fruit or ginseng leaves.

The present inventors applied supercritical extraction and thin film distillation to utilize ginsenosides contained in ginseng fruit and ginseng leaves. As a result, compared with the case where supercritical extraction and thin film distillation were not applied, , The yield was not decreased, and the present invention was completed.

Korean Patent Publication No. 10-2010-0043725 Korea Patent No. 10-0852138 Korean Patent No. 10-1350433 US 5,660,832

There has been a problem in that it is difficult to utilize a crop protection agent when the plant or plant-derived extract is used. It is an object of the present invention to provide a method for removing residual pesticides in a plant comprising the steps of:

1) extracting a plant with a supercritical fluid to separate the lipid soluble component from the plant; And

2) Thin film distillation of the fat soluble component obtained in step 1).

In order to solve the above problems, the present invention provides a method for removing residual pesticide in a plant comprising the steps of:

 1) extracting a plant with a supercritical fluid to separate the lipid soluble component from the plant; And

 2) Thin film distillation of the fat soluble component obtained in step 1).

In addition, the present invention provides a plant extract from which pesticide residues produced by the above method have been removed.

Hereinafter, the present invention will be described in detail.

The present invention provides a method for removing residual pesticides in a plant comprising the steps of:

 1) extracting a plant with a supercritical fluid to separate the lipid soluble component from the plant; And

 2) Thin film distillation of the fat soluble component obtained in step 1).

In addition, the present invention provides a plant extract prepared by the above method.

The method may further comprise the step of pretreating the plant prior to step 1). For example, the fruit of ginseng can be irradiated with any one selected from the group consisting of infrared rays, visible rays and ultraviolet rays. Or the ginseng fruit may be dried. The irradiated or dried ginseng fruit may undergo a steaming / cooling process. The steaming may be carried out at a steam of 70 to 120 캜, preferably at a steam of 90 to 100 캜 for 30 minutes to 5 hours, preferably for 1 to 2 hours, and is cooled after the steaming. This process may be performed once, but is not limited to this. The ginseng fruit that has been steamed / cooled is dried and pulverized to have a water content of 5 to 30%, and the pulverized plant powder is used as an extracting raw material. In case of ginseng leaves, the ginseng leaves are subjected to hot air drying at 70 to 120 ° C, preferably 90 to 100 ° C for 30 minutes to 5 hours, preferably 1 to 2 hours, so that the moisture content of ginseng leaves becomes 10 to 15% Dried and then pulverized, and the powder of the ground ginseng leaves is used as an extraction raw material.

The pesticides disclosed in Table 4 below, except for cypermethrin, metallaclass, pendimethalin, pyraclostrobin and pyrimethanil, can be removed by pretreatment of the plants.

In the step 1), the raw material may be mixed with an organic solvent in the supercritical extraction, and an alcohol, preferably a C ₁ -C ₄ alcohol, may be used as an extraction solvent as an organic solvent that can be used. Accordingly, organic solvents other than alcohols can be used. The organic solvent may be, but is not limited to, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate and mixtures thereof.

In the step 1), the mixing ratio of the raw material and the organic solvent may be 1 kg: 10 to 1 kg: 20 liters, preferably 1 kg: 13 liters to 1 kg: 16 liters, more preferably 1 kg : 13.5 liters to 1 kg: 15.5 liters, but is not limited thereto. If the mixing ratio of the raw material and the organic solvent is less than the lower limit value, that is, if the amount of the raw material is large, the viscosity of the mixture becomes high so that it can not efficiently come into contact with the critical state carbon dioxide and the yield becomes low. When the mixing ratio of the raw material and the organic solvent exceeds the upper limit value, mixing / contact with carbon dioxide is easy because of low viscosity. However, since the amount of the raw materials present in the mixture is small and the yield is low, There are many hassles that must be done several times.

In general, the extraction of water takes place in accordance with the supercritical extraction, and the raw material is made to coalesce due to the decrease of the moisture content, and the handling workability is significantly lowered. Therefore, in the supercritical extraction process, the contact and mixing properties with the supercritical fluid, preferably with the supercritical carbon dioxide, are sufficiently enhanced, and the extraction is progressed efficiently in a short time. At the same time, in order to smoothly carry out the handling work of the ginseng extract, It is preferable to control the mixing ratio of the solvent within the above range and perform the extraction in the low viscosity state.

In the above step 1), in the supercritical extraction, the pressure can be maintained at 200 to 700 bar, preferably 250 to 650 bar, more preferably 300 to 500 bar, but is not limited thereto . When the pressure condition is maintained below the lower limit value, the yield of the fat soluble component decreases even if the extraction time is prolonged. If the pressure condition is maintained above the upper limit value, the production cost is unnecessarily required to maintain the high pressure.

In the above step 1), the supercritical extraction can be carried out at 10 to 40 ° C, preferably at 15 to 35 ° C, more preferably at 20 to 30 ° C , But not limited to. If the value is less than the lower limit value, the extraction yield is lowered. If the upper limit value is exceeded, the production cost is increased compared to the case where the yield is increased, and the cost of the product is increased.

In one embodiment of the present invention, the ginseng fruit powder or the ginseng leaf powder is mixed with ethanol, the mixture is put into an extraction tank, a supercritical fluid such as carbon dioxide is supplied, and then the pressure is maintained at 300 to 500 bar, For 4 hours to obtain 10-20 g of ginseng fruit extract.

During the thin film distillation in the step 2), the internal temperature of the distiller may be 130 ° C to 170 ° C, preferably 135 ° C to 165 ° C, more preferably 140 ° C to 160 ° C But is not limited to, When the internal temperature of the distiller is lower than the lower limit, the pesticide component is not distilled, and the pesticide still remains even though the film is distilled. In addition, when the internal temperature of the distiller exceeds the upper limit value, the pesticide ingredient is removed, but the yield is also lowered because the extract is distilled.

In the thin film distillation in the step 2), the degree of vacuum may be 0.01 torr to 0.5 torr, preferably 0.01 torr to 0.25 torr, and more preferably 0.01 torr to 0.1 torr, but is not limited thereto. When the degree of vacuum is less than the lower limit value, the pesticide component remains without distillation, and when the degree of vacuum exceeds the upper limit value, the pesticide component and the oil component are also distilled together.

The residual pesticide may be selected from the group consisting of cypermethrin, metal lacquer, pendimethalin, pyraclostrobin and pyrimethanil, but is not limited thereto.

The plant may be ginseng, and the ginseng may be any one selected from the group consisting of fine roots of ginseng, root, main root, root, stem, leaf, flower, fruit and fruit, Leaf and ginseng fruit, but is not limited thereto.

The ginseng extract prepared by the method of the present invention can be effectively used in the field of pharmaceutical composition, cosmetic composition or health functional food containing the extract of the present invention or the extract of the present invention because the crop protection agent component is effectively removed.

The ginsenoside content of the ginseng fruit extract or the ginseng leaf extract according to the present invention is maintained or increased and the crop protection agent component is removed compared to the case where the supercritical extraction and the thin film distillation are not applied, Can be utilized in the field of functional foods and cosmetic compositions containing ginseng oil as an active ingredient.

Hereinafter, the present invention will be described in detail with reference to Production Examples, Examples and Experimental Examples.

However, the following Production Examples, Examples and Experimental Examples are for illustrating the present invention, and the contents of the present invention are not limited by the following Production Examples, Examples and Experimental Examples.

< Manufacturing example  1> Ginseng fruit and ginseng leaf pretreatment

<1-1> Pretreatment of ginseng fruit

Ginseng fruit was taken from ginseng and the surface of ginseng fruit was washed with purified water to remove foreign matter. After that, the ginseng fruit was dried, steamed at 98 ° C for 1 hour, cooled, and then dried and pulverized so that the water content of the ginseng fruit was 10 to 15%. The pulverized ginseng fruit powder was used as an extraction raw material Respectively.

<1-2> Pretreatment of ginseng leaves

Ginseng leaves were collected from ginseng and the surfaces of ginseng leaves were washed with purified water to remove foreign matter. Then, the ginseng leaves were dried by hot air at 98 ° C for 1 hour, dried and pulverized to a moisture content of 10 to 15% of the ginseng leaves, and the pulverized ginseng leaf powder was used as an extracting material.

< Example  1> Residual pesticide analysis of ginseng fruit and ginseng leaves

Residual pesticides were extracted from 5 g of the ginseng fruit powder obtained in Production Example 1-1 or the ginseng leaf powder obtained in Production Example 1-2, and the residual pesticide content of the hexane solution used for pesticide removal was investigated.

As the extraction solvent, 90 ml of acetonitrile was used, 10 to 20 g of NaCl was added, sealed and shaken for 2 minutes. After centrifugation at 4 ° C for 5 minutes at 3000 r / min using a centrifuge, 15 ml of the separated acetonitrile layer (supernatant) was taken. The obtained supernatant was concentrated by evaporation until a small amount of solvent was left while passing air or nitrogen gas through a water bath at 40 DEG C or less. The concentrate was purified by eluting with a solution of methylene chloride dissolved in the top of the cartridge.

The extract is divided into GC-MS / MS and HPLC-MS / MS according to the pretreatment method. The eluate was evaporated by passing nitrogen or air through a water bath at 40 ° C or lower at a low flow rate and then redissolved in 2 ml of hexane containing 20% acetone and used as a GC-MS / MS instrument analysis sample. Further, the extract was redissolved in 2 ml of methanol and filtered (0.2 탆) and used as an HPLC-MS / MS analytical sample. Table 1 and Table 2 show the analysis conditions of GC-MS / MS and HPLC-MS / MS. The residual pesticide content after the analysis is shown together with Experimental Example 1 below. Table 4 shows the names of pesticides used for screening.

Analysis conditions of GC-TQ-MSD (Gas Chromatograph Quadrupole Mass Spectrometer) Detector MS Injection temperature 280 ℃ Detector temperature 300 ° C flux 0.8 ml / min column DB-5MS (30 m x 0.25 mm x 0.25 m) Oven temperature program 70 ° C, 3 minutes → 20 ° C / minute → 180 ° C → 5 ° C / minute → 300 ° C, 2 minutes

Analysis conditions of UPLC-TQ-MSD (ultra-fast liquid chromatograph) Detector MS column BEX-C ₁₈ (2.5 x 100 mm, 17 m) flux 0.4 ml / min Mobile phase 0.1% CH ₂ O ₂ / water: 0.1% CH ₂ O ₂ / acetonitrile

UPLC-TQ-MSD Mobile Program Time (minutes) 0.1% CH ₂ O ₂ / water (%) 0.1% CH ₂ O ₂ / acetonitrile (%) Early 80 20 1.00 55 45 6.00 20 80 8.50 20 80 8.60 0 100 9.00 0 100 9.50 80 20 12.00 80 20

< Experimental Example  1> Ginseng leaf, ginseng fruit Supercritical  extraction

The supercritical extraction was carried out at a room temperature for 4 hours while maintaining the pressure in the extraction tank at 450 bar after feeding a mixture of 1 kg of the ginseng powder of Preparation 1-1 and 15 L of ethanol into the supercritical extraction tank and supplying supercritical carbon dioxide . The extract was centrifuged at low temperature and concentrated under reduced pressure to obtain 20 g of a ginseng fruit oil extract (yield: 17%). In case of ginseng leaf powder, 20 g of ginseng leaf oil extract was obtained (yield: 16%), except that ginseng leaf powder of Preparation Example 1-2 was used as a raw material.

< Comparative Example  1> Supercritical  Confirm yield difference by temperature and time during extraction

Supercritical extraction was carried out in the same manner as in Experimental Example 1 except that the conditions of temperature, pressure and time varied. Specific conditions are shown in Table 5 below.

Temperature (℃) Pressure (bar) Time (hr) yield(%) Comparative Example 1-1 40 250 3-5 5 Comparative Example 1-2 45 300 3-5 6 Comparative Example 1-3 50 350 3-5 15 Comparative Example 1-4 55 400 3-5 17 Comparative Example 1-5 65 450 3-5 20 Comparative Example 1-6 70 500 3-5 22 Comparative Example 1-7 75 550 3-5 22 Comparative Example 1-8 80 600 3-5 20

< Experimental Example  2> Thin film distillation

The respective extracts obtained in Experimental Example 1 were distilled using a thin film distiller (UIC KD-6, Germany). The specific distillation conditions are as follows.

1. Flow rate: 2 ml / min

2. Distiller internal temperature 150 ℃, external temperature 170 ℃

3. Vacuum degree 0.06 torr

After the thin film distillation, the residual pesticide and ginsenoside amount were measured by the methods of Examples 1 and 2, and the results are shown in Table 6 below. ND in Table 6 below indicates no detection.

Degree of pesticide residue removal from ginseng fruit oil
Crop protection agent component Cypermethrin Metal rack room Fendi Metalllin Pyraclostrobin Pyrimethanyl etc Ginseng fruit oil 0.21 0.38 0.58 2.89 8.51 ND Ginseng berries
Oil (after thin film distillation)
ND
ND
ND
ND
ND
ND

Degree of pesticide removal of ginseng leaf oil
Crop protection agent component Cypermethrin Metal rack room Fendi Metalllin Pyraclostrobin Pyrimethanyl etc Ginseng leaf
oil 0.34 0.22 0.43 0.21 0.11 ND Ginseng leaf
Oil (after thin film distillation)
ND
ND
ND
ND
ND
ND

As a result of thin film distillation, cypermethrin, metal lacquer, pendimethalin, pyraclostrobin, and pyrimethanil which were detected before thin film distillation were not detected after thin film distillation. The yield of ginseng fruit oil was 95% and the yield of ginseng leaf oil was 96%.

< Comparative Example  2> Evaluation of yield and residual pesticide degree according to thin film distillation conditions

The distillation was carried out in the same manner as in Experimental Example 2 except that the inner temperature and the degree of vacuum were different at the time of thin film distillation, using the ginseng fruit extract of Experimental Example 1. The specific distillation conditions are shown in Table 8 below, and the degree of pesticide remaining after distillation is shown in Table 9.

Internal temperature (℃) Vacuum degree (torr) yield(%) Comparative Example 2-1 60 0.5 97 Comparative Example 2-2 80 0.3 97 Comparative Example 2-3 90 0.08 98 Comparative Example 2-4 120 0.05 97 Comparative Example 2-5 140 0.03 97 Comparative Example 2-6 170 0.01 98

Crop protection agent component yield(%) Cypermethrin Metal rack room Fendi Metalllin Pyraclostrobin Pyrimethanyl etc Ginseng fruit oil 0.21 0.38 0.58 2.89 8.51 ND Experimental Example 2 ND ND ND ND ND ND 95 Comparative Example 2-1 ND ND ND ND 5.51 ND 95 Comparative Example 2-2 ND ND ND ND 3.51 ND 93 Comparative Example 2-3 ND ND ND ND 2.51 ND 91 Comparative Example 2-4 ND ND ND ND 2.11 ND 90 Comparative Example 2-5 ND ND ND ND 1.2 ND 89 Comparative Example 2-6 ND ND ND ND ND ND 87

As a result, it was confirmed that when the temperature inside the distiller was changed in the distillation conditions, the degree of the residual pesticide component and the yield of the ginseng fruit oil were varied. When the internal temperature of the distiller exceeds 220 ° C., the pesticide component is removed, but the ginseng oil is also distilled together to reduce the yield. When the internal temperature is lower than 150 ° C., the pesticide component may not be distilled.

When the degree of vacuum in the degree of vacuum exceeds 0.5 torr, the pesticide component is removed, but the yield of the ginseng fruit oil is also distilled to be low, and when the degree of internal vacuum is less than 0.01 torr, the pesticide component is not distilled Lt; / RTI &gt;

The results of Experimental Example 2 and Comparative Example 2 show that the yield of the ginseng fruit oil is maintained while removing residual pesticides by controlling the conditions of the internal temperature and the vacuum degree of the distiller during the thin film distillation.

Claims (12)

1) extracting the ginseng with supercritical fluid to separate the lipid-soluble component from the ginseng; And
2) A method for removing residual pesticide from ginseng comprising the step of thin-film distilling the fat soluble component obtained in step 1)
Wherein the distiller internal temperature during the thin film distillation in step 2) is in the range of 150 to 170 DEG C and the degree of vacuum is in the range of 0.01 to 0.5 torr. The method according to claim 1,
Further comprising a pretreatment step of selecting ginseng from the group consisting of drying, steaming, and cooling before the step 1). The method of claim 2,
Wherein the steaming is carried out at a temperature of 70 to 120 DEG C for 30 minutes to 5 hours. The method according to claim 1,
Wherein the supercritical fluid is carbon dioxide. delete delete The method according to claim 1,
Wherein the residual pesticide is selected from the group consisting of cypermethrin, metal lacquer, pendimethalin, pyraclostrobin and pyrimethanil. delete The method according to claim 1,
Wherein the ginseng is selected from the group consisting of fine roots, main roots, main roots, pearls, stems, leaves, flower buds, flowers and fruits of ginseng. The method of claim 9,
Wherein the ginseng is at least one of ginseng leaf and ginseng fruit. A ginseng extract obtained by removing the residual pesticide produced by the method of claim 1. delete