WO2013002455A1

WO2013002455A1 - Standard marker for distinguishing country of origin or age of processed ginseng, establishing method thereof, or method for distinguishing country of origin or age of ginseng using same

Info

Publication number: WO2013002455A1
Application number: PCT/KR2011/007368
Authority: WO
Inventors: 오세량; 이형규; 송혁환; 김두영; 김정한; 김정희; 안경섭
Original assignee: 한국생명공학연구원
Priority date: 2011-06-30
Filing date: 2011-10-05
Publication date: 2013-01-03
Also published as: KR101445303B1; KR20130003401A

Abstract

The present invention relates to a standard marker for distinguishing the country of origin or age of processed ginseng, an establishing method thereof, or a method for distinguishing the country of origin or age of ginseng using the same. According to the present invention, the invention is capable of establishing a standard marker for processed Korean ginseng, that is, white ginseng and red ginseng, and can be useful for distinguishing the country of origin or age of the processed ginseng when the processed ginseng is imported and exported by quickly comparing and analyzing the country of origin or age of the processed ginseng from concentrate or powder and the like, for which the country of the origin or age is not easily distinguishable, using the marker.

Description

【Specification】

[Name of invention]

Standard marker for determining the origin or age of processed ginseng, its establishment method or its origin or age determination method

Technical Field

The present invention relates to a standard marker for determining the origin or age of processed ginseng, a method for establishing the same or a method for determining the origin or age using the same.

<2>

Background Art

<3> Conventional non-destructive techniques such as near-infrared spectroscopy, electronic nose, and X-ray fluorescence spectroscopy have been widely used for the determination of the origin of agricultural products. , Milk and dairy products and fats, proteins and solids have been widely used for analysis, and conventional protein analysis methods such as electrophoresis and HPLC analysis can be used to determine the origin or origin of agricultural products, but it is time-consuming and expensive. There was a disadvantage in that it was difficult to apply in the field. Recently, however, the CE chromatogram has been applied as a new method that can minimize the time and sample amount required for analysis because of the shorter analysis time and higher resolution than conventional methods, and enables quantitative digital analysis. In particular, it is mainly used to discriminate wheat varieties (Kim Nam-su, "Development of discrimination technology and safety assessment technology for domestic and imported agricultural products (processed foods)" research project report, 2003).

<4> Therefore, scientific discrimination is made to effectively classify the difference in quality caused by the origin and cultivation conditions of various agricultural products including processed ginseng and unprocessed ginseng, and to reinforce the labeling and institutionalization of agricultural products by the WT0 system. As the development of methods is urgently needed, and there is an increasing number of imported agricultural products which are illegally distributed, the development of scientific identification methods for the verification of the origin of these agricultural products collected in the process of on-site enforcement is required.

<5>

Ginseng (panax ginseng CA Meyer) has been used as a medicinal plant for thousands of years, and ginseng has long evolved to suit its environment. Little by little, he has been able to live well in his environment and has changed himself according to the environment, and ginseng is divided into fresh ginseng, white ginseng, and heungsam dong according to the processing method, and the ginseng of mined natural state is ginseng. Peeled ginseng and dried in sunlight or hot air is called white ginseng. Also, white ginseng is steamed without peeling. Steamed and dried is called red ginseng.

<7> Korean ginseng has been known to be the best in Korea. In recent years, however, international markets have been encroached by cheap Chinese and American and Canadian ginseng, and Chinese ginseng has recently entered the domestic market through wheat trade, making it difficult to position domestically grown ginseng. Study on the Establishment and Efficacy of Classification System of Wild Ginseng and Chinese Camphor Ginseng "Research Project Report, 2007), Also, overseas ginseng does not have much meaning for lotus root discrimination because it is mined 3-4 years old ginseng. According to the conventional belief that the older roots are more effective, the older roots are more expensive because of the higher price. Lotus root discrimination is a big problem.

<8>

<9> The current inspection criteria for the determination of lotus root by the Ministry of Agriculture, Forestry and Welfare is determined by the inspecting staff at the time of harvest, and is determined by visual or color development of the head, trunk and epidermis, the degree of leg development, and the age at the time of cutting. (Ministry of Agriculture, Ginseng Industry Law, Article 8, 2004), which is used as an auxiliary discrimination method at the site where the cultivation history is confirmed, but it is controversial due to the level of sensory evaluation, and moreover, the distribution history is unclear. It is difficult to apply in the market. The reported lotus root discrimination method has been the cerebral cranial scar (J. Ginseng Res. Vol.31, 142-146, 2007), and the ring (J. Ginseng Res. Vol 30, 153-157, 2006, Ginseng Research Report , Korean Ginseng and Tobacco Research Institute, p331, 1987), Geun Dal (Ginseng Research Report-Cultivation, Korean Ginseng and Tobacco Research Institute, pl75, 1988), Machine drying method (J. Ginseng Res. Vol 30, 153-157, 2006) Tissue staining (J. Ginseng Res. Vol 25, 101-105, 2001), Ginsenoside content (Can. J. Plant Sci. Vol 76, p 855 1996) Assay The other methods have low reliability and are used as auxiliary. However, the secretory vascular staining method is limited to cultivated ginseng and is not applicable to ginseng powder, and it is difficult to apply to processed red ginseng because it is distinguished by root section of root.

In order to solve the above problems, the present inventors separate and analyze standard markers from Korean ginseng and Chinese ginseng, which are studying the origin or lotus root discrimination method of Korean ginseng and Chinese ginseng. The present invention was completed by confirming that it is possible to determine a mountainous region or lotus root. <11>

[Content of invention]

[Technical problem]

An object of the present invention is to provide a marker for producing white ginseng mountain tube discrimination used for mountain production of white ginseng.

Another object of the present invention is to provide a method for determining the production of white ginseng.

Another object of the present invention is to provide a marker for determining the red ginseng mountain region used for mountain determination of Heungsam.

Another object of the present invention is to provide a method for determining a mountain of Heungsam.

Another object of the present invention to provide a marker for determining the age of Heungsam used for determining the age of Heungsam.

Another object of the present invention is to provide a method for determining the age of red ginseng.

<18>

Technical Solution

In order to achieve the above object, the present invention provides a marker for determining the origin of white ginseng represented by the following Chemical Formula 1 or 2, which is used to determine the origin of domestic white ginseng and Chinese white ginseng:

<20> [Formula 1]

<22> [Formula 2]

In addition, the present invention comprises the steps of extracting a metabolite from the test white ginseng (step 1);

Analyzing the metabolite extracted in step 1 by liquid chromatography-mass spectrometry (LC-MS) (step 2); And

It provides a method for determining the production of white ginseng comprising the step (step 3) comparing the LC-MS analysis obtained in step 2 with the marker for determining the mountain region of Formula 1 or 2. Furthermore, the present invention provides a marker for determining a mountain area of Heungsam represented by the following Chemical Formula 3 or 4, which is used for mountain production of domestic red ginseng and Chinese heungsam:

[Formula 3]

In addition, the present invention comprises the steps of extracting a metabolite from the test red ginseng (step 1);

It provides a method for determining a mountain of Heungsam, comprising the step (step 3) of comparing the LC-MS analysis result obtained in step 2 with the mountain determination marker of the formula (3) or (4). Furthermore, the present invention provides a marker for determining the age of red ginseng represented by the following Chemical Formulas 5 to 11 used for the lotus root discrimination of domestic four-year-old red ginseng and six-year-old red ginseng:

<36> [Formula 5]

<38> [Formula 6]

<47> [Formula 10]

<49>

In addition, the present invention comprises the steps of extracting the metabolite from the test heungsam (step 1);

Comparing the LC-MS analysis result obtained in step 2 with the age determination marker selected from the group consisting of the compounds represented by the formula (5 to 11) (step 3) age determination method of red ginseng To provide.

<54>

Advantageous Effects

According to the present invention, a standard marker is established in domestically processed ginseng, that is, white ginseng and red ginseng, and the origin or age of processed ginseng such as concentrate or powder, which is difficult to identify the origin or age, can be quickly determined using the marker. As it is possible to compare and analyze, it can be useful for import and export of ginseng seedlings for determination of origin or age.

<56>

[Brief Description of Drawings]

Figure 1 shows an ion chromatogram of domestic white ginseng extract of the embodiment of the present invention I am drawing. '

2 is a diagram showing an ion chromatogram of Chinese ginseng extract of the embodiment of the present invention.

FIG. 3 is a diagram illustrating a core scatter plot of domestic white ginseng extract and Chinese white ginseng extract of an embodiment of the present invention.

Figure 4 is a view showing the S plot of the least orthogonal square analysis of domestic white ginseng extract and Chinese white ginseng extract of the embodiment of the present invention.

FIG. 5 is a diagram illustrating a box plot of (a) domestic white ginseng extract and (b) Chinese white ginseng extract of an embodiment of the present invention.

6 is a view showing a mass cutting method of the domestic white ginseng extract of the embodiment.

7 is a view showing a mass cutting method of the Chinese white ginseng extract of the embodiment.

8 is a diagram showing an ion chromatogram of a domestic Heungsam extract of an embodiment of the present invention.

9 is a view showing an ion chromatogram of Chinese red ginseng extract of the embodiment of the present invention.

10 is a view of the domestic heungsam extract and Chinese heungsam extract of the embodiment of the present invention

Is a diagram showing a core scatter plott.

FIG. 11 is a view showing an S plot of the least orthogonal square analysis of domestic red ginseng extract and Chinese red ginseng extract according to an embodiment of the present invention.

FIG. 12 is a diagram illustrating a box plot of (a) Korean red ginseng extract and (b) Heungsam extract made in China according to an embodiment of the present invention.

Figure 13 is a view showing the mass cutting method of the domestic red ginseng extract of the embodiment.

14 is a view showing a mass cutting method of the Chinese heungsam extract of the embodiment.

FIG. 15 is a diagram illustrating an ion chromatogram of a domestic four-year-old Heungsam extract of an example of the present invention.

FIG. 16 is a diagram illustrating an ion chromatogram of Korean six-year-old red ginseng extract of the example of the present invention. FIG.

17 is a four-year-old Heungsam extract and six-year-old red ginseng extract of the domestic embodiment of the present invention . The score scatter plot of FIG.

FIG. 18 is a diagram illustrating an S-plot of the least-orthogonal square analysis of domestic 4-year-old Heungsam extract and 6-year-old Red ginseng extract according to an embodiment of the present invention.

FIG. 19 is a diagram illustrating a box plot of the domestic 4-year-old Heungsam extract of the embodiment of the present invention. FIG.

20 is a view showing a box plot of the domestic six-year-old red ginseng extract of the embodiment of the present invention.

<77>

[Best form for implementation of the invention]

Hereinafter, the present invention will be described in detail.

<79>

The present invention provides a marker for determining the production site of white ginseng represented by the following formula (1) or 2 used for determining the production of domestic white ginseng and Chinese white ginseng:

<8)> [Formula 1]

<83>

At this time, the compound represented by the formula (1) is a marker for determining the origin of domestic white ginseng And, the compound represented by the formula (2) is a marker for determining the origin of Chinese ginseng.

<85>

Marker for determining the production site of white ginseng represented by the formula (1) or 2 according to the present invention comprises the steps of extracting a metabolite from domestic white ginseng and Chinese white ginseng (step 1);

In step 2, the result obtained by converting the result analyzed by LC—MS into a numerical value that can be statistically processed, and analyzing the converted value by multivariate analysis, detects a component that shows a difference between groups, and shows the largest difference. Can be established in a method comprising the step of selecting and establishing the component as a standard marker according to the region of origin (step 3).

<89>

Specifically, step 1 according to the present invention is a step of extracting metabolites from domestic white ginseng and Chinese white ginseng, and the extraction solvent is preferably water, alcohol or a mixture thereof. As the alcohol, it is preferable to use d to C ₄ lower alcohol, more preferably to use ethanol or methanol, and most preferably to use 70% methane, but not always limited thereto. As the extraction method, conventional methods in the art, such as filtration, hot water extraction, dipping extraction, reflux angle extraction, and ultrasonic extraction, can be used, and the extraction is preferably performed once to five times by ultrasonic extraction, and repeated three times. More preferably, but not limited to extraction. The extraction temperature is preferably 20 to 30 ° C, but is not limited thereto.

<91>

In addition, step 2 is a liquid chromatography of the metabolite extracted in step 1

Column chromatography was used at 30-40 ^° C., where the mobile phase A was water and formic acid (100: 0.1, v / v), and the mobile phase B was analyzed by mass spectrometry (LC-MS). Was used as acetonitrile (acetonitril) and formic acid (100: 0.1, v / v). The flow rate of the mobile phase in minutes and the sample injection amount can be used for measurement by taking 1 sample of domestic white ginseng extract and 1 sample of Chinese white ginseng extract. In addition, the mass spectrometer may use a quadrupole orthogonal accelerated flight time tandem mass spectrometer, but is not limited thereto.

<93>

Further, step 3 converts the result analyzed by LC-MS into a numerical value capable of statistical processing, and analyzes the converted value by multivariate analysis to determine the difference between the data and the group. After detecting the visible components, selecting the components with the highest difference to establish standard markers of domestic white ginseng and Chinese white ginseng as standard markers according to the regions. The multivariate analysis method used here is an orthogonal least square analysis method. The data obtained by analyzing the multivariate method can be used to identify standard markers using a score scatter plot.

<95>

In addition, the present invention

Extracting the metabolite from the test white ginseng (step 1);

It provides a method for determining the production of white ginseng comprising the step (step 3) comparing the LC-MS analysis result obtained in step 2 with the marker for determining the mountain region of Formula 1 or 2.

<100>

The white acid production method of white ginseng can be used to determine the origin of white ginseng during import and export, since it is possible to quickly compare white ginseng production areas such as concentrates and powders that are difficult to identify.

<102>

Furthermore, the present invention provides a marker for determining the production site of red ginseng represented by the following Chemical Formula 3 or 4, which is used to determine the production of domestic Heungsam and Chinese Heungsam:

<104> [Formula 3]

[Formula ⁴ ]

At this time, the compound represented by the formula (3) is a marker for discriminating the production site of Korean red ginseng, the compound represented by the formula (4) is a marker for discriminating the production site of Chinese red ginseng. In addition, the marker for determining the production site of red ginseng represented by the formula (3) or 4 according to the present invention comprises the steps of extracting a metabolite from domestic heungsam and Chinese heungsam (step 1);

The result analyzed by LC-MS in step 2 is converted into a numerical value that can be statistically processed, and the component having the difference between the groups is detected from the data obtained by analyzing the converted value by multivariate analysis, and the component having the largest difference is selected. By establishing a standard marker according to the mountain region (step 3).

Specifically, the steps 1 to 3 may be carried out in the same manner as the mountain irrigation method of the white ginseng, but is not limited thereto. In addition, the present invention

Extracting the metabolite from the test heungsam (step 1);

It provides a method for determining the production location of red ginseng comprising the step (step 3) comparing the LC-MS analysis obtained in step 2 with the mountain determination marker of the formula (3) or (4). The production method of the red ginseng can be quickly compared with the use of standard markers, such as concentrated powder, which is difficult to identify the production area using a standard marker, so the import and export of red ginseng It can be useful for discrimination. Furthermore, the present invention provides a marker for determining the age of red ginseng represented by the following Chemical Formulas 5 to 11 used for the discrimination of lotus root of four-year-old Heungsam and six-year-old Korean red ginseng:

[Formula 5]

5> [Formula 8]

<129>^'

At this time, the compound represented by the formula 5 to 8 is a marker for determining the age of 4 years old red ginseng, the compound represented by the formula 9 to 11 is a marker for determining the age of 6 years old red ginseng. ^'

<131>

The age-determining marker of red ginseng represented by Chemical Formulas 5 to 11 according to the present invention includes the steps of extracting metabolites from domestic 4-year-old Heungsam and 6-year-old Heungsam (step 1);

In step 2, the result analyzed by LC-MS is converted into a numerical value capable of being statistically processed, and the transformed value is analyzed by multivariate analysis to detect components showing differences between groups, and the largest difference is found. It can be established by a method comprising the step of selecting the ingredients and establishing them as standard markers according to the age of Heungsam.

Specifically, the steps 1 to 3 may be performed in the same manner as the mountain determination method of the white ginseng, but is not limited thereto.

<136>

In addition, the present invention,

Extracting the metabolite from the test heungsam (step 1);

Analyzing the metabolite extracted in step 1 by liquid chromatography-mass spectrometry (LC ᅳ MS) (step 2); And

The method of determining age of Heungsam comprising the step of comparing the LC-MS analysis result obtained in step 2 with an age discriminating marker selected from the group consisting of the compounds represented by Chemical Formulas 5 to 11 (step 3). To provide.

<141>

The age determination method of red ginseng can be usefully used to determine the age of red ginseng at the time of import and export, since it is possible to quickly compare the age of Heungsam such as concentrated liquid and powder, which are difficult to identify by using a standard marker.

<143>

Furthermore, the present invention provides a kit for determining a mountain of white ginseng comprising a marker represented by Chemical Formula 1 or 2.

In addition, the present invention provides a kit for determining the location of red ginseng comprising a marker represented by the formula (3) or (4). Furthermore, the present invention provides a kit for determining the lotus root of domestic red ginseng comprising any one or more of the markers represented by the formulas (5) to (11).

<147>

Kits prepared according to the present invention can be usefully used to determine the origin of white ginseng or red ginseng, or to determine the lotus root of domestically grown ginseng.

<149>

[Form for implementation of invention]

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

<152>

<153> <Manufacture example 1> Preparation of processed ginseng

<154> Domestic White Ginseng for the Identification of Korean and Chinese White Ginseng regardless of Lotus Root (age)

53 points were purchased from Korea, 18 white ginseng from China were purchased locally and in China, and 50 red ginseng ginseng from Korea were purchased from Korea to determine the origin of Korea and Heungsam in China. Purchased 35 points at was used in the experiment. For the sake of discrimination of 4 and 6 roots of red ginseng, red ginseng was prepared from 4 root 10 roots and 6 roots 10 roots of ginseng from Icheon, Gyeonggi-do.

<155>

Example 1 Establishment of a Standard Marker for Determination of Mountain Production of White Ginseng

The following experiments were conducted to establish markers for determining the origin of domestic white ginseng and Chinese white ginseng.

Step 1: Extraction of White Ginseng Metabolites

<15,9> After drying the domestic white ginseng and Chinese white ginseng prepared in Preparation Example 1, and crushed 0.2 mg each of each, add 5 ml of 70% methane, and irradiated with 350W ultrasonic wave for 1 hour ， The extract was filtered through a 0.2 μιη membrane filter (SUN-S i, TN, USA) and the extracts were collected.

<160>

Step 2: Particle Extract of White Ginseng Extract Using Liquid Chromatography-Mass Spectrometry (LC-MS)

In order to analyze the metabolites of the white ginseng extract prepared in step 1, UPLC used the Waters Acquisition UPLC System, and the column was the Equity ™ BEHC ₁₈ (ACQUITY). UPLC ™ BEHCi ₈ ) (10 隱 X2.1 mm, id, 1.7ym, waters, USA) was used at 35 t, mobile phase A for water and formic acid (100: 0.1, v / v) and mobile phase B for aceto Initially increase mobile phase B from 10% to 30% for 7 minutes with nitrile (acetonitril) and formic acid (100: 0.1, v / v) and then move to mobile phase B up to 14 minutes at 44% and 21 minutes at 100%. After increasing the analysis, the mobile phase B was maintained at 10 to 23. minutes, the mobile phase B was reduced to 10% at 23.6 minutes, and after equilibrating, it was used for the next analysis. The flow rate of the mobile phase was 0.4 minutes, and the sample injection amount of each sample of domestic white ginseng extract and Chinese white ginseng extract was used for each measurement.

<i63> The Q-T0F mass spectrometer used Waters' quadrupole orthogonal accelerat ion time of—flight tandem mass spectrometer (Waters Q-T0F Premier ™). Analysis conditions of the mass spectrometer are shown in Table 1 below.

<164> [Table 1]

<165>

The total chromatogram of liquid chromatography-mass spectrometry (LC-MS) of the domestic white ginseng extract and the Chinese white ginseng extract is shown in FIGS. 1 and 2.

<167>

Step 3: Transformation into Statistical Processable Numbers and Establishment of the White Ginseng Standard Marker

In order to convert the LC-MS data obtained in step 2 into numerical values capable of being statistically processed, peak finding and peak sorting are performed using a Markersynx applications manager (ver. 4.1) of Waters Corporation. peak alignment and peak filtering were performed. For the above process, 0.5 to 19 minutes of the analytical data were used, and the analytical mass range was 200-1500 Da, mass The tolerance range was 0.02 Da, and the mass noise level was 1。 of the base peak intensity. In all LC-MS data obtained through this process, the retention time (RT) value of each peak coincides with the mass value, and the area value of each peak is obtained accordingly, and then Simca-P + ver for multivariate statistical analysis. Using the 12.0 (Umetrics, Sweden) software, all data were par-scaled (par-scaled (scaled to square root of SD)) to group domestic and Chinese white ginseng to score the orthogonal least squares analysis (0PLS-DA). Cat plots are shown, and S plots (Simca-P +) of orthogonal least-squares analyzes are used to select natural peaks that can be distinguished by box plots of components with high differences between the two mountain regions and their mass ion chromatograms [ Mass ion chromatogram, MassLynx (ver 4.1)] was used to identify structures that differed significantly by selecting standard markers by mass fragmentation pattern. The results are shown in FIGS. 3 to 7.

<170>

<171> results

(1) As a result of the score scatter plot of the orthogonal least-squares analysis, as shown in FIG. 3, domestic white ginseng and Chinese white ginseng were separated into two groups according to the mountain region.

(2) In addition, as a result of the S plot of the orthogonal least squares analysis, as shown in FIGS. 4 and 5, a component having a mass of 963.5582 for 5.21 and a mass having 771.4919 for 9.30 is significant. There was a difference (t-test; p> 0.001).

(3) Furthermore, as a result of the mass fragmentation pattern, as shown in FIGS. 6 and 7, the standard marker of domestic white ginseng is 20-glucosinenoside Rf ( 5.21 minutes, MW 963.5582), the standard marker of Chinese white ginseng, as shown in the following formula 2, notojincetoside. R2 (9.30 min, M.W. 771.4919).

<175>

Example 2 Establishment of a Standard Marker for the Production of Red Ginseng

The following experiments were conducted to establish markers for determining the origin of domestic Heungsam and Chinese red ginseng.

Step 1: Extraction of Red Ginseng Metabolites

Except for using domestic red ginseng and Chinese red ginseng prepared in Preparation Example 1 was carried out in the same manner as in Step 1 of Example 1. Step 2: Analysis of Red Ginseng Extract Using Liquid Chromatography-Mass Spectrometry (LC-MS)

Ά

Except for using the metabolite of the red ginseng extract prepared in step 1 was carried out in the same manner as in step 2 of Example 1.

Results of the total ion chromatogram of the liquid chromatography-mass spectrometry (LC-MS) of the domestic Heungsam extract and the Chinese Heungsam extract are shown in FIGS. 8 and 9. <I89>

Step 3: Statistical Processing and Establishment of Red Ginseng Standard Markers

<i9i> The same process as in Example 3, except that the results obtained in Step 2 were used.

<192>

<193> result

(1) As a result of the score scatter plot of the orthogonal least-squares analysis, as shown in FIG. 10, domestic red ginseng and red ginseng red ginseng were separated into two groups according to the mountain region.

(2) In addition, as a result of the S plot of the orthogonal least-squares analysis, as shown in FIGS. 11 and 12, there is a significant difference between a component having a mass of 801.4988 / 9.04 and a component having a mass of 767.4952 / 11.70. (T-test; p> 0.001).

(3) Furthermore, the mass fragmentation pattern result, FIG. 13 and

As shown in 14, the standard marker of Korean red ginseng was identified as ginsenoside Rf (9.04 minutes, MW 801.4988), as shown in Formula 3 below, and the standard marker of Heungsam in China was shown in Formula 4 below. Ginsenoside Rd (11.70 minutes, MW 767.4952).

<197>

<198> [Formula 3]

<199>

<200> [Formula 4]

<202>

Example 6 Establishment of a Standard Marker for Determining the Age (4 Years, 6 Years) of Red Ginseng

The following experiments were conducted to establish markers for the determination of mountainous regions of domestic 4-year-old Heungsam and 6-year-old Heungsam.

Step 1: Extraction of Red Ginseng Metabolites

It was performed in the same manner as in Step 1 of Example 1 except for using the four-year-old domestic red ginseng and 6-year-old domestic red ginseng prepared in Preparation Example 1.

<207>

Step 2: Determination of Red Ginseng Extract Using Liquid Chromatography-Mass Spectrometry (LC-MS)

Except for using the metabolite of the Heungsam extract prepared in step 1 was carried out in the same manner as in step 2 of Example 1.

<2io> Total chromatogram results of liquid chromatography-mass spectrometry (LC-MS) of the domestic 4-year-old Heungsam extract and 6-year-old red ginseng extract are shown in FIGS. 15 and 16.

<211>

Step 3: Transformation into Statistical Processable Numbers and Establishment of Red Ginseng Standard Markers

The same procedure as in Example 3 was performed except that the result obtained in Step 2 was used.

<2] 4>

<215> result

(1) As a result of the score scatter plot of the orthogonal least squares analysis, as shown in FIG. 17, domestic four-year-old red ginseng and six-year-old red ginseng were divided into two groups according to lotus root. (2) Moreover, the S plot result of the orthogonal least squares analysis, shown in FIGS. 18 to 20 As shown, the components of 4-year-old Heungsam and 6-year-old Heungsam showed significant differences in mass (t-test; p> 0.001).

<2i8> Of these, four-year-old heungsam component is malonyl ginsenoside Rbl (1195.6145 / 9.89 minutes) (Formula 5), malonyl ginsenoside Rc (10.30 minutes, MW 1165.6030) (Formula 6), malonyl ginsenosides Side Rb2 (10.75 minutes, MW 1165.6036) (Formula 7), malonyl ginsenoside Rd (12.02 minutes, MW 853.4980) (formula 8) was confirmed, and the six-year-old Heungsam component is (R) -ginsenoside Rhl ( 10.48 min MW 1277.8934) (Formula 9), Queen Quenoside RK 11.26 min, MW 1151.6239) (Formula 10), (R) -ginsenoside Rg3 (15.80 minutes, M.W. 749.4871) (Formula 11).

The four-year-old red ginseng component and the six-year-old heungsam component are shown in Table 2 below.

<220> [Table 2]

<221> <222>

Claims

[Claim of Claim] [Claim 11] A marker for determining the production location of white ginseng represented by the following Chemical Formula 1 or 2, which is used for determination of production of domestic white ginseng and Chinese white ginseng.

[Formula 1]

[Claim 2]

According to claim 1, wherein the compound represented by the formula (1) is a marker for determining the production of white ginseng, characterized in that the marker for determining the production of domestic white ginseng.

[Claim 3]

According to claim 1, wherein the compound represented by the formula (2) is a marker for determining the production of white ginseng, characterized in that the marker for determining the production of white ginseng from China.

[Claim 4]

Extracting metabolites from domestic white ginseng and Chinese white ginseng (step 1); Analyzing the metabolite extracted in step 1 by liquid chromatography-mass spectrometry (LC-MS) (step 2); And

The result analyzed by LC-MS in step 2 is converted into a numerical value that can be processed statistically, and the component having the difference between the groups is detected from the data obtained by analyzing the converted value by multivariate analysis, and the component having the largest difference is selected. Method of establishing a marker for determining white mountain production of claim 1, comprising the step (step 3) of establishing as a standard marker according to the mountain.

[Claim 5]

5. The method according to claim 4, wherein the multivariate analysis method of step 3 is an orthogonal least square analysis method.

[Claim 6]

5. The method of claim 4, wherein the data obtained by analyzing the multivariate analysis of step 3 is in a score scatter plot.

[Claim 7]

Extracting the metabolite from the test white ginseng (step 1);

Comparing the results of the LC-MS analysis obtained in step 2 with the marker for determining the mountain region of Chemical Formula 1 or 2 of claim 1 (step 3).

[Claim 8]

Marker for determining the location of red ginseng represented by the following formula (3) or (4), which is used to determine the production of domestic red ginseng and Chinese heungsam. [Formula 3]

[Claim 9]

The method of claim 8, wherein the compound represented by the formula (3) is a marker for determining the production site of Heungsam, characterized in that the marker for determining the production of Korean red ginseng.

[Claim 10]

■ The marker according to claim 8, wherein the compound represented by Chemical Formula 4 is a marker for determining the origin of Chinese red ginseng.

[Claim 111]

Extracting metabolites from domestic red ginseng and Chinese heungssam (step 1);

In step 2, the result obtained by converting the result analyzed by LC-MS into a numerical value that can be statistically processed, and showing the difference between the data from the data obtained by analyzing the converted value by the multivariate analysis method. The method of claim 8, wherein the step of detecting a component, selecting the component having the largest difference and establishing it as a standard marker according to the production region (step 3).

[Claim 12]

12. The method of claim 11, wherein the multivariate analysis method of step 3 is an orthogonal least-squares analysis method.

[Claim 13]

12. The method according to claim 11, wherein the data obtained by analyzing the multivariate analysis of step 3 is a score scatter plot.

[Claim 14]

Extracting the metabolite from the test red ginseng (step 1);

Comparing the results of the LC-MS analysis obtained in step 2 with the acid determination marker of the formula (3) or claim 8 (step 3) of the production area determination of red ginseng.

[Claim 15]

A marker for determining the age of Heungsam represented by the following Chemical Formulas 5 to 11 used for the discrimination of lotus root of domestic Heungsam.

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

30

[Formula 9]

[Claim 16]

16. The marker for determining age of Heungsam according to claim 15, wherein the age of discrimination of Heungsam is 4 years or 6 years.

[Claim 17]

16. The marker for determining age of Heungsam according to claim 15, wherein the compound represented by Chemical Formulas 5 to 8 is an age discriminating marker of 4 years old red ginseng.

[Claim 18]

16. The marker for determining age of red ginseng according to claim 15, wherein the compound represented by Chemical Formulas 9 to 11 is an age discriminating marker of red ginseng 6 years old.

[Claim 19]

Extracting metabolites from domestic four-year-old heungsam and six-year-old heungsam (step 1); Analyzing the metabolite extracted in step 1 by liquid chromatography-mass spectrometry (LC-MS) (step 2); And

The result of LC-MS analysis in step 2 is converted into a numerical value that can be statistically processed, and the component having the difference between groups is detected from the data obtained by analyzing the converted value by multivariate analysis, and the component having the largest difference is selected Method of establishing the red ginseng age discrimination marker according to claim 15 comprising the step (step 3) of establishing as a standard marker according to the age of red ginseng.

[Claim 20]

20. The method according to claim 19, wherein the multivariate analysis method of step 3 is an orthogonal least-squares analysis method.

[Claim 21]

20. The method of claim 19, wherein the data obtained by analyzing the multivariate analysis of step 3 is a score scatter plot.

[Claim 22]

Extracting the metabolite from the test heungsam (step 1);

Heungsam age determination method comprising the step (step 3) comparing the LC-MS analysis result obtained in step 2 with an age discriminating marker selected from the group consisting of compounds represented by the formulas 5 to 11 of claim 15 .

[Claim 23] Kit for determining the production of white ginseng comprising a marker represented by the formula (1) or (2) of claim 1. [Claim 24]

Kit for determining the production of red ginseng comprising a marker represented by the formula (3) or (4) of claim 18. [Claim 25]

A kit for irrigation of lotus root in domestic Heungsam, comprising any one or more of the markers represented by Chemical Formulas 5 to 11.