KR102145406B1 - Method for analyzing effective ingredients in ginseng - Google Patents

Abstract

The present invention relates to a method for measuring active ingredients of ginseng, characterized by comprising the step of analyzing the content of active ingredients of ginseng by a near-infrared reflection spectrum, and according to the present invention, chemical treatment or processing of ginseng, processed ginseng products, or ginseng extract It is possible to accurately and quickly measure the content of active ingredients of ginseng contained in it even without doing so.

Description

Method for analyzing effective ingredients in ginseng}

The present invention relates to a method for measuring active ingredients of ginseng, and more particularly, to a method for rapidly measuring the content of main active ingredients contained in ginseng without chemically altering or processing ginseng.

Ginseng ( Panax ginseng ) is a plant that grows in deep mountainous areas and is commonly cultivated for medicinal purposes. Saponin glycosides, such as josponin and ginsenoside, which are known to be the main ingredients of ginseng medicinal properties, have anti-fatigue, work ability enhancement, sex gland growth promotion, and blood sugar-lowering effects. The above efficacy, represented by an adaptogen effect, in particular enhances the non-specific immunity of the living body.

Due to the excellent efficacy of ginseng, health functional foods made from ginseng are continuously being released, and efforts to quantitatively indicate their quality are also continuously being made. In order to liquefy the main ingredients of ginseng by extraction, refining, pulverization, etc., to select the quality of the health functional foods sold in pouches, etc., and to provide a basis for reasonable judgment for consumers, the Korea Food and Drug Administration The content of Rb1+Rg1, which is the sum of ginsenoside Rb1 and ginsenoside Rg1, and the content of crude saponin, which are the main ingredients of the medicinal effect, were established as criteria for quality evaluation of processed ginseng products.

Typically, the measurement of the components contained in ginseng has been made through a chemical calibration method such as high performance liquid chromatography (HPLC), including grinding and processing. It is a statistical method to determine the composition ratio of the entire product line by setting up several representative groups among a large number of product groups and conducting chemical composition evaluation for them. However, the above method not only includes the demands of a professional manpower and procedural complexity, but also requires a long period of time and is highly inefficient in terms of cost.

The problem to be solved by the present invention conceived to overcome the above limitations is a simple method that can quickly measure the content of ginseng active ingredients contained in ginseng, processed ginseng products, or ginseng extracts without chemical treatment or processing. To provide a way.

The present invention in order to achieve the above object,

b) analyzing the content of the active ingredient in ginseng using a near-infrared reflection spectrum;

It provides a method of measuring ginseng active ingredient comprising a.

According to an embodiment of the present invention, before step b),

a) Calibration showing the correlation between the active ingredient content of ginseng and the near-infrared reflection spectrum by comparing the content of a plurality of active ingredients of ginseng for calibration analyzed by the component measurement method including HPLC method and the near-infrared reflection spectrum of the ginseng for calibration Setting the formula or calibration data;

It may further include.

According to another embodiment of the present invention,

Step b) may be performed based on a calibration formula or calibration data.

According to another embodiment of the present invention,

Step b),

After irradiating the ginseng with near-infrared rays, a reflection spectrum is obtained, and the obtained reflection spectrum may be subjected to water treatment using a differential function, multiple scattering correction, or both.

According to another embodiment of the present invention,

The derivative of step b) includes a first derivative, a second derivative, or both, the segment of the first derivative or the second derivative is 5 units or 7 units, and the smoothness is 3, 7 Or it could be 9.

According to another embodiment of the present invention,

Step a),

After irradiating the ginseng with near-infrared rays, a reflection spectrum is obtained, and the obtained reflection spectrum may be subjected to water treatment using a differential function, multiple scattering correction, or both.

According to another embodiment of the present invention,

The derivative of step a) includes a first derivative, a second derivative, or both, the segment of the first derivative or the second derivative is 5 units or 7 units, and the smoothness is 3, 7 Or it could be 9.

According to another embodiment of the present invention,

The wavelength of the near infrared ray may be 950-1650 nm.

According to another embodiment of the present invention,

The near-infrared reflection spectrum may be selected in a wavelength range of 1110-1170, 1300-1360, 1370-1430, or 1470-1530 nm.

According to another embodiment of the present invention,

The active ingredient may be irradiation ponin, ginsenoside Rb1 or ginsenoside Rg1.

According to another embodiment of the present invention,

The ginseng may be white ginseng, fresh ginseng, or red ginseng.

According to another embodiment of the present invention,

The active ingredient is irradiated phonin,

The near-infrared reflection spectrum is statistically processed as one point set in a multidimensional space,

The calibration formula may be Equation 1 below.

[Equation 1]

y=(0.60 to 0.70)x + (0.30 to 0.45)

According to another embodiment of the present invention,

The active ingredient is ginsenoside Rb1,

The near-infrared reflection spectrum is statistically processed as one point set in a multidimensional space,

The calibration formula may be the following formula 2.

[Equation 2]

y=(0.83 to 0.92)x + (12 to 25)

According to another embodiment of the present invention,

The active ingredient is ginsenoside Rg1,

The near-infrared reflection spectrum is statistically processed as one point set in a multidimensional space,

The calibration formula may be Equation 3 below.

[Equation 3]

y=(0.92 to 0.98)x + (0.65 to 0.85)

As described above, according to the present invention, it is possible to accurately and quickly measure the content of ginseng active ingredients contained in ginseng, processed ginseng products, or ginseng extracts without chemical treatment or processing.

1 is a graph showing the measurement of the near-infrared reflection spectrum of a ginsenoside Rb1 standard substance that appears differently according to a concentration gradient.
FIG. 2 is a graph showing a near-infrared reflection spectrum of a ginsenoside Rb1 standard substance, which appears different according to a concentration gradient, by first water treatment.
Figure 3 is a graph showing the measurement of the near-infrared reflection spectrum of the ginsenoside Rg1 standard substance that appears differently according to the concentration gradient.
FIG. 4 is a graph showing a near-infrared reflection spectrum of a ginsenoside Rg1 standard substance, which appears different according to a concentration gradient, by first water treatment.
Figure 5 is a ginsenoside Rb1 and ginsenoside Rg1 in which a concentration gradient (0.1, 0.01, 0.001% by weight) is formed, and a principle component analysis (PCA) of one commercially available ginseng pouch product is shown. It is a graph.
6 is a graph showing a reflection spectrum obtained after irradiating a plurality of commercial ginseng pouch products with near-infrared rays.
7 is a graph showing a reflection spectrum obtained after irradiation with near-infrared rays on a plurality of commercially available ginseng pouch products by primary water treatment.
8 is a graph of the calibration equation of ginsenoside Rb1 derived from the near-infrared reflection spectrum of a commercial ginseng pouch product.
9 is a graph of the calibration equation of ginsenoside Rg1 derived from the near-infrared reflection spectrum of a commercial ginseng pouch product.
10 is a graph of the calibration formula of irradiated ponins produced from the near-infrared reflection spectrum of a commercial ginseng pouch product.

Hereinafter, the present invention will be described in detail.

The present invention is ginseng ('ginseng' described in the specification of the present invention should be understood as a concept including all of ginseng processed products such as fresh ginseng, red ginseng, white ginseng, and commercially available ginseng extract pouch products) The ginseng effect according to the present invention was developed with attention to the fact that the reflection spectrum of Near Infra-Red (NIR) significantly changes according to the content of ginsenoside Rb1, ginsenoside Rg1 and irradiated ponins, which are the main components of The component measurement method is characterized in that the content of the active ingredient contained in ginseng is analyzed using a near-infrared reflection spectrum.

Near-infrared rays exist at 700-2,500 nm, between branched rays and mid-infrared rays, and have lower energy than visible rays and higher energy than mid-infrared rays. Near-infrared spectroscopy takes advantage of the fact that most natural products absorb near-infrared radiation in a specific region or wavelength. In particular, NH, O-H and C-H bonds strongly absorb near-infrared radiation, while other molecules are weakly absorbed. Absorption of near-infrared rays appears mainly in the bonding band and harmonic band of the basic molecular vibration energy of the N-H, O-H and C-H bonds existing in the mid-infrared rays. In the near-infrared rays appearing in the bonding band and the harmonic band, the absorbance is very weak. It can be seen that the near-infrared (NIR) band is mainly used for quantitative analysis of the sample, and the mid-infrared (MIR) band is used for quantitative analysis.

Since the absorbance energy of near-infrared rays is 10 to 1,000 times smaller than the basic molecular vibration energy of functional groups derived from mid-infrared rays, it is convenient to measure as it is without the need for pretreatment like mid-infrared rays. In the near-infrared region, the absorption by moisture is very weak, so a total inspection is possible, and the information represented by the functional groups, such as CH, NH, OH, which constitute organic matter, is compressed and displayed in the near-infrared region, so that information on the physicochemical properties of agricultural products can be obtained simultaneously I can. In addition, when a specific wavelength that can express the state of a sample is determined, the spectroscopic analysis method is easier to construct a quality evaluation system compared to other non-destructive testing methods, does not require skilled skills during analysis, and makes the interpretation of the results simple and measurement at low cost. There is an advantage to be able to configure the system.

By comparing the absorbance obtained at a specific wavelength using a near-infrared spectroscopy (NIR), the content of the main ingredient of ginseng can be roughly confirmed, and the sample can be quickly analyzed without any additional pretreatment. Pretreatment (dissolution, heating, homogenization, etc.) of the sample can be minimized by performing analysis using light in the near-infrared region, and variations for each spectrum can be confirmed through treatment (water treatment) using a mathematical function for the spectrum. have. For example, the differential processing of the spectrum makes it easier to distinguish the difference in the absorption spectrum for each sample by making the correction for the base line and the change in the part with the shift larger.

In order to practice the present invention, ginseng does not need to be pulverized, chemically processed, or chromatographed, and the reflection spectrum obtained therefrom after irradiating ginseng with near-infrared rays, which will be described later, includes fine powder and scattering correction. Since the analysis is performed according to the method, and the analysis may be quickly performed by a computerized or mechanized method, the method of measuring the ginseng active ingredient according to the present invention is more useful in testing a large amount of ginseng samples.

In order to accurately quantify and algorithmize the correlation between the near-infrared reflection spectrum and the main component of ginseng, prior to the step of irradiating the ginseng to be measured with near-infrared rays, the method for measuring the active ingredient of ginseng according to the present invention is a conventional method including HPLC. The method may further include the step of obtaining a plurality of near-infrared reflection spectra of ginseng for calibration with the content of the main component analyzed, and setting a calibration formula or calibration data for analyzing a correlation between the content of the main component and the near-infrared reflection spectrum. In the step of setting the calibration formula or calibration data, the PLSR method (Partial Least Squares Regression) of the multivariate regression analysis method may be used to obtain a standard calibration formula.If the calibration formula has been set in the past, this step is performed. It is possible to omit it.

The calibration equation is set up using a quantitative analysis method and Beer's law to obtain a linear equation with the transmittance and reflectivity of the sample to be measured. A known sample is collected and selected to correct the scattering effect of the light irradiated on the sample. After that, regression analysis is performed and a calibration curve is derived, verified, and then applied to routine analysis.

In addition, the "scatter correction" means correcting a nonlinear function that distorts the correlation between the spectrum and the experimental value. Scattering correction methods include the following. (1) SNV (Standard normal variant): The particle size effect is reduced by reducing each spectrum from the baseline to the standard order difference of 1. (2) Detrend: Removes linear or quadratic curvature from each spectrum. (3) Standard MSC (Multiplicative scatter correction): Each spectrum is corrected using an average spectrum.

When measuring the spectrum, changes in the surrounding environment (lighting, temperature, etc.), light scattering on the surface of the object, the size of the object, the difference in distance between the object and the measuring sensor, and noise from the measuring sensor have a significant influence on the measured light spectrum. . Moreover, if the optical spectrum is measured in real time at the agricultural product screening site, not in a laboratory environment, more influence will be exerted. Therefore, this effect affects the spectrum used to grasp the optical characteristics, causing spectral variation irrelevant to the information of internal quality, causing errors in the prediction of internal quality measurement, and acting as a factor affecting the performance of the developed quality judgment. Is done.

In order to correct errors caused by such spectral shift, a spectrum water treatment process is performed. Spectral water treatment is a very basic and important technique that removes noise on optical measurements and reduces errors due to its influence, and is used to obtain more stable optical spectrum characteristics.

The main methods of spectral water treatment are smoothing, MSC, SNV, average and maximum values, three kinds of normalization using a range of values, Savitzky-Golay and Norris Gap methods using first and second differential characteristics. There is this. Among them, the smoothing method is used to remove noise from the spectrum measurement device, and the first and second derivatives are used to remove the shift of the baseline due to differences in optical paths or changes in the measurement environment, or to emphasize the spectral characteristics of minute components. do. In addition, MSC and SNV are used to remove the effect of light scattering during spectrum measurement. The Norris Gap derivative is similar to the Savitzky-Golay derivative, but can be used by adjusting the gap size.

That is, the reflection spectrum obtained in the present invention may include a step of water treatment to reduce the influence due to the scattering effect and facilitate quantification. The water treatment may be performed by differential, multiplicative scatter correction, or both by a predetermined order, and the differentiation may be a first derivative, a second derivative, or both. The differential segment may be 5 units or 7 units, and the smoothness may be 3, 7 or 9.

The method of measuring the active ingredient of ginseng according to the present invention may be characterized in that it is performed using near-infrared rays having a wavelength of 950-1650 nm, among others, 1110-1170, 1300-1360, 1370-1430 or 1470-1530 nm The reflection spectrum selected in the wavelength range can be selected and used for content measurement. Although it will be described in more detail in the examples to be described later, the above wavelength range shows a pattern that reacts sensitively to irradiation phonin, ginsenoside Rb1, or ginsenoside Rg1 contained in ginseng, and the present invention as the above range When performed, the accuracy of the measured content also increases rapidly.

The method of measuring the ginseng active ingredient according to the present invention, as has been described so far, can be said to have been specifically specialized in irradiation phonin, ginsenoside Rb1 and ginsenoside Rg1. However, according to the technical idea described in the present invention, the range of measurable components can be further broadened even if the wavelength domain and reflection spectrum analysis algorithm are slightly changed, and in particular, analysis of the measurable ingredients of various ginseng including ginsedoside Rg3 It will be well known to those skilled in the art to use the present invention.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these examples are for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

Materials and methods

(1) Preparation of active ingredient standards and samples

As a standard material,

Ginsenoside Rb1 (Sigma Aldrich): 0.1% by weight sample (hereinafter, Rb101W.1.1)

Ginsenoside Rb1 (Sigma Aldrich): 0.01% by weight sample (hereinafter, Rb1001W.1.1)

Ginsenoside Rb1 (Sigma Aldrich): 0.001% by weight sample (hereinafter, Rb10001W.1.1)

Ginsenoside Rg1 (Sigma Aldrich): 0.1% by weight sample (hereinafter, Rg101w.1.1)

Ginsenoside Rg1 (Sigma Aldrich): 0.01% by weight sample (hereinafter, Rg1001w.1.1)

Ginsenoside Rg1 (Sigma Aldrich): 0.001% by weight sample (hereinafter, Rg10001w.1.1)

As a sample,

200 commercially available ginseng pouch products (2012 red ginseng 6 years old) were used.

(2) NIR Measure

The NIR spectrum was measured from 950 to 1,650 nm using a DA7200 Analyser (Perten instrument AD, Sweden), and 1 mL of the sample for measurement was absorbed in a glass microfibre filter (GF/A, Whatman Ltd., England) and then absorbed. Measured.

(3) Calibration Chemical measurement of active ingredient content in samples for setup and verification

1) irradiation ponins:

Take the sample into a reflux flask, add hydrated BuOH, cool to reflux for 1 hour in a 80℃ water bath, cool at room temperature, and transfer only the hydrated BuOH layer to a separate funnel. Add distilled water to the collected hydrated BuOH, shake it, and let stand until the water layer and the BuOH layer are completely separated, remove the water layer, and transfer the BuOH layer to a constant-weight flask to completely concentrate. Add ether to the concentrated residue, and after cooling under reflux for 30 minutes in a 46℃ water bath, completely discard the ether and blow off the odor. Put the residue in a dry oven at 105℃, dry it, and allow it to cool for 30 minutes, then weigh it.

Irradiation ponin content (mg/g)=A-B/S

A: The weight of the flask after concentrating and drying the hydrated BuOH layer (mg)

B: Weight of the empty flask as a constant weight (mg)

S: Sample collection amount (g)

2) ginsenosides Rb1 and Rg1:

The contents of ginsenosides Rb1 and Rg1 were analyzed by HPLC. The instrument used at this time consisted of a Jasco (Tokyo, Japan) HPLC with a PU-2089 Plus gradient pump equipped with a degasser, an AS-2075 Plus autosampler, and an MD-2010 Plus multi wavelength detector. The analytical column is Sunfire ^TM C ₁₈ (4.6x250 mm id, 5 µm, Waters, Milford, MA, USA) was used, and the injection volume was 20 µl, the flow rate was 0.8 ㎖/min, the column temperature was 40 ℃, and the absorption wavelength of the UV detector was set to 280 nm. I did. As the mobile phase, (A) 2% acetic acid, (B) 0.5% acetic acid, and 50% acetonitrile were degassed with an ultrasonic cleaner, and a gradient system was used, and based on B, 0% (0 min. ), 55% (70 minutes), 100% (73 minutes), 0% (78 minutes), and 0% (80 minutes).

(4) Method of statistical processing

In the present invention, the NIR analysis uses a method of finding a predicted value by placing the entire spectrum in one multidimensional space and interpolating the position as a single point into a linear equation of y = ax + b using the mathematical least squares method.

If there are three reflection spectrum data, each of them is compressed into one point in a three-dimensional space, and if there are 100 data, each can be expressed as one point in a 100-dimensional space. This mathematically uses a matrix, increasing the order to 3 x 3 for 3D and 100 x 100 for 100D. The task of lowering these data back to the level that the user can read is called PCA (Principal Components Analysis). In general, the main component is compressed into two or three axes.

After compressing the data using this statistical method, the compressed one point is compared with the result of the experiment with the standard experimental method, and a straight line of the least squares method is obtained, which becomes a calibration equation, and becomes a linear equation with slope and intercept.

Predicted values can be obtained by adding compressed spectral data of unknown samples to the set calibration equation. The analysis method using the least-squares method is sometimes used for more accurate calibration even in the analysis method using UV or HPLC that used the conventional 1:1 comparison method. As in the present invention, an analysis method that compares a number of 1: is referred to as a multivariate analysis method.

Test example One: Ginsenoside Rb1 Standard material Near infrared Correlation of reflection spectrum

FIG. 1 is a graph showing by irradiating near-infrared rays on ginsenoside Rb1 standard substances and obtaining the reflection spectrum thereof, and FIG. 2 is a graph showing the reflection spectrum graph of FIG. It is a graph shown.

As shown, the near-infrared reflection spectrum showed a significant difference according to the concentration gradient of 0.1% by weight, 0.01% by weight, and 0.001% by weight, especially 1110-1170, 1300-1360, 1370-1430 or 1470-1530 nm wavelength It can be seen that the difference is more pronounced in the area. This difference can be seen more clearly in the water-treated graph of FIG. 2.

Test example 2: Ginsenoside Rg1 Standard material Near infrared Correlation of reflection spectrum

FIG. 3 is a graph showing by irradiating near-infrared rays on ginsenoside Rg1 standard substances and obtaining a reflection spectrum thereof, and FIG. 4 is a graph showing the reflection spectrum graph of FIG. It is a graph shown.

As shown, the near-infrared reflection spectrum showed a significant difference according to the concentration gradient of 0.1% by weight, 0.01% by weight, and 0.001% by weight, especially 1110-1170, 1300-1360, 1370-1430 or 1470-1530 nm wavelength It can be seen that the difference is more pronounced in the area. This difference can be seen more clearly in the water-treated graph of FIG. 4. The above aspect is very similar to Test Example 1.

Test example 3: Ginsenoside Rb1 And Rg1 Standard and sample Selectivity analysis

FIG. 5 shows the results of principal component analysis (PCA) on a ginsenoside Rb1 standard, a ginsenoside Rg1 standard, and a randomly selected sample 1 individual.

PCA is a principal component analysis, which is a widely used analysis method in statistics. The key is to abbreviate multidimensional data into several main components. With this abbreviated data, it goes through the process of finding only the necessary information. In the case of data containing a lot of information, you can display it in a multidimensional space to display it as a single location. Data compression using this multidimensional space and the axis that can contain the most information of data is drawn. This is the principle component (PC). However, since the first principal component (PC1) does not contain all the data, it is perpendicular to the principal component axis and creates a principal component axis that can hold a lot of information after the first principal component. This is the second main component (PC2). Of course, many principal components such as PC3 and PC4 can be made, but the principal component analysis compresses data and converts data in a multidimensional space into a two-dimensional space (e.g., coordinates between PC1 and PC2 axes or PC1 and PC3 axes) so that humans can intuitively understand them. Because it is showing, if the number of PCs is too large, it moves back to the multidimensional space and interferes with intuitive understanding. The chart of Fig. 5 shown on the axis of this main component is called Score Plot.

In the graph above, the x-axis represents the first principal component PC1, which is computed from the NIR spectrum, and the y-axis represents the second principal component PC2, computed from the NIR spectrum. In the lower left corner (red circle), PC1 contains 99% of the information. That means it accounts for 99%.

Principal components can be grouped differently for different purposes. If you want to work by grouping the data you want to analyze into several groups, adjust the number and axis of PCs so that each data is well grouped. Or, to find the problem, you can find the material that is the most distant from the center of the material. This can eliminate outliers.

In FIG. 5, group separation was induced by using a method of creating groups for each component content derived by computerizing each NIR spectrum. HSJ1w (brown color) is the sample, green represents the standard product by concentration of Rb1, and the sky blue represents standard product by concentration of Rg1. It was concluded that concentration classification by NIR was sufficiently possible as the products were displayed in different spaces for each concentration, and the product also occupied a position in the space for each concentration.

Test example 4: of the sample Ginsenoside Rb1 , Rg1 And Irradiation Chemical analysis of the content

As described above, chemical measurement of the active ingredient content included in 200 samples of commercially available ginseng pouch products was made to set the calibration formula or calibration graph, and the results are shown in Table 1 below.

Sample number Irradiation phonin
(mg/ml) Ginsenoside (μg/ml) Sample number Irradiation phonin
(mg/ml) Ginsenoside (μg/ml) Rg1 Rb1 Rg1 Rb1 One 1.17 18.57 193.53 101 1.13 9.20 89.29 2 1.27 18.93 197.17 102 1.22 9.34 91.07 3 1.28 18.67 193.55 103 1.15 9.87 91.33 4 1.37 19.77 199.32 104 1.37 9.68 92.50 5 1.23 18.92 197.19 105 1.31 9.92 94.29 6 1.26 18.76 198.30 106 1.36 10.92 95.97 7 1.35 26.72 204.85 107 1.23 10.69 94.18 8 1.41 27.68 209.13 108 1.20 9.53 88.87 9 1.38 27.48 210.70 109 1.15 10.72 90.57 10 1.18 26.81 205.96 110 1.28 11.13 93.63 11 1.18 29.20 219.38 111 1.23 9.21 88.13 12 1.16 28.09 214.74 112 1.20 9.04 86.64 13 1.21 26.08 202.93 113 1.32 10.01 89.45 14 1.70 28.59 213.97 114 1.26 10.43 92.20 15 1.05 29.42 220.08 115 1.22 0.00 46.70 16 0.62 6.31 32.86 116 1.12 0.00 54.89 17 0.60 10.07 33.51 117 1.18 0.00 57.14 18 0.76 9.79 33.16 118 1.25 9.66 93.29 19 0.70 10.18 32.08 119 1.08 0.00 50.41 20 0.99 10.03 32.60 120 1.11 0.00 56.04 21 0.68 10.42 32.96 121 0.89 21.08 39.34 22 0.57 8.41 47.08 122 0.96 22.72 39.97 23 0.69 6.91 47.24 123 0.88 22.01 41.01 24 0.65 7.94 46.15 124 0.87 21.98 39.79 25 0.54 8.27 48.15 125 0.86 20.88 41.45 26 0.59 7.76 50.07 126 0.82 20.90 42.26 27 0.76 6.51 47.73 127 0.84 21.92 39.23 28 0.66 7.70 48.55 128 0.92 21.51 37.38 29 0.56 7.36 49.36 129 0.90 22.18 38.16 30 0.59 7.75 48.95 130 0.94 21.97 39.85 31 1.16 12.57 55.84 131 0.92 23.28 40.14 32 1.22 11.03 53.36 132 0.99 28.19 63.26 33 1.08 11.93 54.85 133 0.97 27.91 49.58 34 0.90 11.54 60.24 134 1.00 28.60 64.59 35 0.93 11.31 57.10 135 1.02 28.79 64.77 36 0.99 15.29 56.24 136 0.94 21.81 37.98 37 0.95 7.09 101.60 137 1.06 29.09 60.15 38 0.65 6.87 99.65 138 1.04 21.45 35.62 39 0.93 6.89 102.25 139 1.18 22.37 38.93 40 1.09 7.16 103.69 140 1.09 24.08 44.43 41 0.97 6.41 100.89 141 1.11 24.47 45.02 42 0.77 6.34 100.21 142 1.07 25.23 39.62 43 1.13 6.43 99.67 143 1.07 24.07 44.58 44 1.07 6.26 98.14 144 1.05 23.80 36.15 45 1.18 6.34 97.23 145 1.00 21.74 33.37 46 0.90 44.68 224.91 146 1.07 22.51 34.07 47 1.13 46.08 228.59 147 1.06 21.97 41.43 48 1.02 45.15 227.50 148 1.03 22.92 35.13 49 1.50 45.67 225.62 149 0.99 21.73 41.41 50 0.99 46.35 214.62 150 1.01 23.07 37.99 51 1.20 33.08 165.49 151 1.08 21.26 35.49 52 1.19 44.50 176.74 152 0.97 21.24 35.10 53 1.07 44.95 173.56 153 1.00 20.82 50.96 54 0.81 45.35 171.60 154 1.05 22.48 38.42 55 1.24 45.34 170.46 155 0.96 22.36 33.49 56 1.22 47.28 179.73 156 0.97 21.65 32.25 57 0.75 47.39 173.95 157 0.99 21.42 36.77 58 0.79 47.91 179.36 158 0.97 21.60 50.80 59 1.32 47.93 172.06 159 1.03 21.17 50.10 60 1.47 47.29 173.65 160 1.01 21.47 50.81 61 1.15 3.34 19.02 161 1.05 21.50 100.99 62 1.18 2.78 15.81 162 1.23 19.89 61.19 63 1.18 13.47 71.95 163 0.97 21.40 85.48 64 1.15 6.46 37.13 164 1.09 20.59 77.11 65 1.21 10.54 58.74 165 1.04 21.76 86.44 66 1.14 9.33 53.55 166 1.16 21.83 61.32 67 1.22 11.15 64.24 167 1.16 21.96 87.01 68 1.26 10.05 58.83 168 0.98 22.63 65.80 69 1.25 5.67 34.86 169 1.04 22.30 62.03 70 1.15 4.47 26.85 170 0.99 22.19 61.52 71 1.20 7.44 44.20 171 1.07 22.05 60.27 72 1.10 6.12 37.79 172 1.05 20.95 64.85 73 1.12 7.53 44.34 173 1.07 21.16 62.74 74 1.17 3.41 20.26 174 0.97 21.80 65.45 75 1.13 5.66 32.72 175 1.03 21.01 62.29 76 1.13 11.83 35.06 176 1.01 20.58 61.32 77 1.20 12.71 67.86 177 1.05 20.75 63.00 78 1.22 10.28 57.21 178 1.00 21.05 61.77 79 1.03 9.35 52.33 179 1.01 20.83 63.54 80 1.09 13.13 74.25 180 1.01 21.10 63.25 81 1.35 12.12 70.53 181 1.23 7.37 44.74 82 1.20 9.00 53.22 182 1.19 8.86 44.99 83 1.12 5.55 32.97 183 1.22 2.55 45.85 84 1.08 3.54 21.91 184 1.29 8.43 44.57 85 1.00 4.50 26.27 185 1.32 7.93 46.92 86 0.99 6.20 39.21 186 1.29 5.98 44.48 87 1.26 2.93 17.10 187 1.21 2.59 45.30 88 1.14 3.91 22.82 188 1.20 2.32 45.08 89 1.02 11.28 66.58 189 1.27 6.98 44.64 90 1.10 10.32 59.62 190 1.27 7.43 45.11 91 0.94 11.71 100.86 191 1.42 2.41 46.11 92 1.08 11.27 97.70 192 1.33 2.96 44.73 93 1.20 10.46 94.93 193 1.39 7.52 45.67 94 1.16 10.97 90.44 194 1.33 6.87 45.08 95 1.07 9.60 89.09 195 1.35 5.58 45.26 96 1.14 9.95 90.23 196 1.32 6.42 44.99 97 1.04 9.60 88.82 197 1.36 6.80 45.10 98 1.16 9.29 87.73 198 1.34 7.14 44.92 99 1.08 9.88 92.44 199 1.37 1.85 45.22 100 1.19 9.19 87.54 200 1.31 3.64 70.60

Example 1: of the sample Near infrared Reflection spectrum measurement

FIG. 6 is a graph showing by irradiating near-infrared rays on ginsenoside Rg1 standard substances and obtaining the reflection spectrum thereof, and FIG. 7 is a graph showing the reflection spectrum graph of FIG. It is a graph shown.

As shown, there was a significant difference in the near-infrared reflection spectrum depending on the sample, and in particular, it can be seen that the difference is more pronounced in the 1110-1170, 1300-1360, 1370-1430 or 1470-1530 nm wavelength region. This difference can be seen more clearly in the water-treated graph of FIG. 7. The above aspect is very similar to Test Example 1 and Test Example 2.

Example 2: Near infrared analysis Calibration Settings

Hereinafter, the calibration set and verification set referred to in the examples below include a chemical measurement amount and an NIR analysis amount performed on the samples. However, the calibration set is the set used when making the calibration equation regressed to the least squares method (PLS). The software Unscrambler (ver. 9.7, CAMO Software AS, Norway) used here shows all data available for verification through Cross Validation.

The blue regression equation (calibration set) included in the diagrams of the following examples is a calibration equation regressed through calibration. The red regression equation (validation set) is a calibrated calibration equation created by substituting all data into the created calibration equation through cross validation for verification.

Definitions of terms used in the following examples are as follows.

Slope: The slope of each calibration or validation set made with PLS. The closer to 1, the better.

Offset: The point (intercept) of the regression equation that intersects the Y coordinate (predicted value). The closer to 0, the better.

RMSE: As the average difference between the measured and predicted values for the calibration set and validation set, RMSE can be interpreted as the average error value of the model and has the same unit as the original response value. The closer to 0, the better.

R-Square: The square value of the correlation coefficient between the measured value and the predicted value as a measure of the quality of the model in the calibration model. This value is always between 0 and 1, and the higher the better.

(One) Ginsenoside Rb1 Near infrared analysis Calibration Settings

Through the software Unscrambler described above, the correlation between the ginsenoside Rb1 content (Table 1) contained in the sample and the water-treated NIR reflection spectrum of the sample (Fig. 7) was analyzed and a calibration equation was set (Fig. 8). The results are shown in Table 2 below.

division inclination Offset RMSE R-Square Calibration set (Rb1) 0.9095 13.20 22.66 0.9095 Validation set (Rb1) 0.8378 23.88 34.35 0.8013

(2) of the sample Ginsenoside Rg1 Near infrared analysis Calibration Settings

Through the software Unscrambler described above, the correlation between the ginsenoside Rg1 content (Table 1) contained in the sample and the water-treated NIR reflection spectrum of the sample (Fig. 7) was analyzed and a calibration equation was set (Fig. 9). The results are shown in Table 3 below.

division inclination Offset RMSE R-Square Calibration set (Rg1) 0.9669 0.74 0.37 0.9669 Validation set (Rg1) 0.6693 2.45 1.48 0.5053

(3) of the sample Irradiation Near infrared analysis Calibration Settings

Through the software Unscrambler described above, the correlation between the irradiation phonin content contained in the sample (Table 1) and the water-treated NIR reflection spectrum of the sample (FIG. 7) was analyzed and a calibration equation was set (FIG. 10). The results are shown in Table 4 below.

division inclination Offset RMSE R-Square Calibration set (Izoponin) 0.6974 0.34 0.12 0.6974 Verification Set (Izoponin) 0.6251 0.42 0.15 0.5661

Example 3: The chemical measured amount of the active ingredient in the sample and NIR Comparison of reflection spectrum analysis amount

Based on the calibration formula for ginsenoside Rb1, ginsenoside Rg1, and irradiation ponin set in Example 2, NIR content analysis was performed on 13 randomly selected samples, and the results are shown in Table 5 below. .

Sample number Irradiation phonin (R ² =0.7864) Rb ₁ (R ² =0.8924) Rg ₁ (R ² =0.9373) NIR analysis value
(mg/mL) Chemical Analysis Value
(mg/mL) NIR analysis value
(mg/mL) Chemical Analysis Value
(mg/mL) NIR analysis value
(mg/mL) Chemical Analysis Value
(mg/mL) Random 1 1.00 1.16 46.6 40.2 22.4 20.2 Random 2 1.46 1.22 48.9 44.4 30.5 38.4 Random 3 1.12 1.08 49.1 49.8 27.4 22.4 Random 4 0.98 0.90 46.3 56.1 26.4 24.1 Random 5 0.89 0.93 72.4 66.4 27.2 34.1 Random 6 1.15 0.99 48.3 55.9 40.3 42.4 Random 7 1.25 0.95 65.4 70.2 33.6 40.1 Random 8 1.01 0.65 56.3 57.9 67.1 74.8 Random 9 0.89 0.93 77.4 70.4 43.6 40.3 Random 10 1.7 0.97 79.5 83.6 56.4 49.8 Random 11 1.94 1.45 87.1 78.9 66.9 60.2 Random 12 1.91 1.78 89.4 69.4 69.1 61.9 Random 13 1.91 1.42 98.2 106.7 59.4 51.8

As indicated above, according to the present invention, it is possible to reliably analyze the active ingredient content of ginseng in a remarkably short time only by comparing a simple near-infrared irradiation and a set calibration formula.

Claims (14)

b) After irradiating the ginseng with near-infrared rays, a reflection spectrum was obtained, the obtained reflection spectrum was water-treated using a differential function, multiple scattering correction, or both, and the content of the active ingredient of ginseng was determined using the water-treated near-infrared reflection spectrum. Including; analyzing;
The near-infrared reflection spectrum is selected from a wavelength range of 1110-1170, 1300-1360, 1370-1430 or 1470-1530 nm,
The derivative includes a first derivative, a second derivative, or both, the segment of the first derivative or the second derivative is 5 units or 7 units, and the smooth (smooth) is 3, 7 or 9,
The active ingredients are ginsenoside Rb1 and ginsenoside Rg1,
The content analysis in step b) is performed based on a calibration equation, and the near-infrared reflection spectrum is statistically processed as one point set in a multidimensional space,
The calibration formula of the ginsenoside Rb1 is the following formula 2, and the calibration formula of the ginsenoside Rg1 is the measuring method of ginseng active ingredient, characterized in that the formula 3;
[Equation 2]
y=(0.83 to 0.92)x + (12 to 25)
[Equation 3]
y=(0.92 to 0.98)x + (0.65 to 0.85)
In Equations 2 and 3, x is a water-treated near-infrared reflection spectrum, y in Equation 2 is the content of ginsenoside Rb1, and in Equation 3, y is the content of ginsenoside Rg1. The method of claim 1, before step b),
a) Calibration showing the correlation between the active ingredient content of ginseng and the near-infrared reflection spectrum by comparing the content of a plurality of active ingredients of ginseng for calibration analyzed by the component measurement method including HPLC method and the near-infrared reflection spectrum of the ginseng for calibration Setting the formula or calibration data;
Measuring method of ginseng active ingredient, characterized in that it further comprises. delete delete delete The method of claim 2,
Step a),
After irradiating ginseng with near-infrared rays, a reflection spectrum is obtained, and the obtained reflection spectrum is water-treated using a differential function, multiple scattering correction, or both. The method of claim 6,
The derivative includes a first derivative, a second derivative, or both, the segment of the first derivative or the second derivative is 5 units or 7 units, and the smoothness is 3, 7 or 9 Method for measuring active ingredient of ginseng characterized by. The method of claim 2,
The measuring method of ginseng active ingredient, characterized in that the wavelength of the near infrared is 950-1650 nm. delete delete The method according to any one of claims 1 to 2,
The ginseng is white ginseng, fresh ginseng or red ginseng, characterized in that the measuring method of ginseng active ingredient. delete delete delete

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