KR100817633B1 - Method for distinguishing healthy peppers from diseased peppers using solvent free solid injector mounted gas chromatography - Google Patents

Abstract

A method for distinguishing health peppers from diseased peppers is provided to avoid a solvent extraction step for pepper samples, thereby simplifying the process and improving the accuracy in analysis, and to prevent diseases in peppers. A method for distinguishing health peppers from diseased peppers by using a solvent-free solid sample injector mounted to a gas chromatography system, comprises the steps of: mounting an injector(120) of a solvent-free solid sample injection system(100) to an injector port(210) provided in the gas chromatography system(200); inserting a glass capillary(150) in which a predetermined amount of pepper sample(160) is sealed into the injector; preheating the injector to vaporize the pepper sample; breaking the glass capillary to guide the vaporized pepper sample to the column(230) of the gas chromatography system; analyzing the fragrance in the vaporized pepper sample; and determining whether an indole compound exists in the fragrance or not, and regarding pepper samples containing indole compounds as diseases peppers and pepper samples not containing indole compounds as healthy peppers.

Description

METHODS FOR DISTINGUISHING HEALTHY PEPPERS FROM DISEASED PEPPERS USING SOLVENT FREE SOLID INJECTOR MOUNTED GAS CHROMATOGRAPHY}

1 is a perspective view of a solvent-free solid sample injector used in the present invention.

2A to 2C are schematic diagrams illustrating a method of analyzing aroma components of red pepper using a gas chromatograph equipped with a solvent-free solid sample injector used in the present invention.

3 is a graph showing the effect of the preheating temperature of the injector to detect the flavor component of the pepper.

Figure 4 is a graph showing the effect of the preheating time of the injector to detect the flavor component of the pepper.

5 is a graph showing the effect of the time to induce the vaporized pepper sample to the column of the gas chromatograph on the flavor component detection of pepper.

<Description of the symbols for the main parts of the drawings>

100: solvent-free solid sample injector 110: support

120: injector 125: outlet of the vaporized sample

130: glass capillary mill 140: adjustment screw

150: glass capillary 160: pepper sample

200: gas chromatograph 210: injector port

220: heating means 230: column

240: analysis means

The present invention relates to a method for distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph. More specifically, the injector of the solvent-free solid sample injector is mounted on the injector port provided in the gas chromatograph. After mounting a glass capillary tube in which a predetermined amount of capsicum samples are sealed in the injector, the injector is preheated to vaporize the sample sealed therein, and the glass capillary is crushed to vaporize the capsicum sample in the gas chromatograph. After deriving the column, by analyzing the fragrance component of the pepper sample, if it contains indoles, it is judged as two-pepper pepper, and if it does not contain indole-type distinction between two peppers and healthy peppers judged as whole pepper Solvent-free solid sample mounted on gas chromatograph showing how to It relates to a method of distinguishing healthy and diseased chili pepper with flag.

In analyzing the fragrance components, Simulaneous steam distilation & extraction, Supercritical Fluid Extraction (SFE), Accelerated Solvent Extraction (ASE), Solid Phase Microextraction (Solid Phase Microextraction); SPME) and Purge & Trap method.

The co-distillation and extraction method is the most common and has been used for a long time as the extraction method by distilled water, the extraction of the fragrance is often used organic solvents rather than distilled water.

The supercritical fluid extraction method is a method of using a supercritical fluid at a temperature and pressure higher than a critical temperature and a critical pressure, and easily penetrates into a solid sample to extract analyte in a short time.

The accelerated solvent extraction method is a method of extracting a substance in a short time using a small amount of an organic solvent and a water-soluble solvent in a solid or semi-solid state of the sample at elevated temperature and pressure.

The solid phase trace extraction method is a method in which the sample liquid is directly adsorbed onto silica, desorbed by heat, and then moved inside the column of the chromatograph.

The Purge & Trap method is a method in which a liquid or solid sample is bubbled by an inert gas, and then a volatilized component is adsorbed to a trap and rapidly desorbs volatile components by heat into an injector of a chromatograph.

On the other hand, conventionally, when determining whether the pepper is a diseased pepper or diseased healthy pepper not diseased pepper was distinguished between the healthy pepper or two diseased pepper through the fragrance component analysis method.

However, the analytical methods have a disadvantage of mainly undergoing an extraction process and a purification process in order to analyze the fragrance component of red pepper. That is, since extraction is performed by using a mixture of water and an organic solvent at the time of extraction, not only two or three solvent extraction processes to remove water are required but also a purification process for removing impurities is required.

Therefore, there is a problem that the analysis process is complicated, there is a problem that it is difficult to accurately determine the healthy peppers and two diseased peppers because the flavor component of the pepper is lost and difficult to accurately analyze.

Therefore, the present invention is to solve all the disadvantages and problems of the prior art as described above, not only go through the solvent extraction process of pepper samples, but also simple and fast as well as to accurately analyze the flavor components without loss of flavor components and healthy peppers The purpose is to provide a way to accurately distinguish this disease pepper.

In addition, an object of the present invention is to provide a method for accurately distinguishing the initial pepper anthrax by analyzing the fragrance components of healthy peppers and two diseased peppers to enable early control of pepper anthrax.

The object of the present invention is the step of mounting the injector of the solvent-free solid sample injector to the injector port provided in the gas chromatograph; Inserting a glass capillary sealed with a predetermined amount of pepper samples into the injector of the solvent-free solid sample injector; Preheating the injector to vaporize the pepper sample sealed therein; Breaking the glass capillary to guide the vaporized pepper sample to a column of the gas chromatograph; Analyzing the fragrance components of the evaporated chili pepper sample; And judging whether or not indole is contained in the fragrance component of the red pepper sample, when indoles are included, judging as two red peppers, and judging as healthy red peppers when indoles are not included. It is achieved by a method of distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph.

In addition, the object of the present invention is a dance equipped with a gas chromatograph, characterized in that the injector of the solvent-free solid sample injector is provided with a plurality of holes to guide the vaporized pepper sample to the column of the gas chromatograph It is also achieved by the method of distinguishing healthy and diseased peppers from each solid sample injector.

In addition, the above object of the present invention is also achieved by a method for distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph, wherein the preheating temperature of the injector is 200 to 300 ° C.

In addition, the above object of the present invention is also achieved by a method for distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph, wherein the preheating temperature is 250 ° C.

In addition, the above object of the present invention is also achieved by a method for distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph, wherein the preheating time of the injector is 3 to 10 minutes.

In addition, the above object of the present invention is also achieved by a method for distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph, wherein the preheating time is 7 minutes.

In addition, the object of the present invention is the time to induce the vaporized sample to the column of the gas chromatograph is 10 minutes or less, characterized in that the whole red pepper and two red pepper using a solvent-free solid sample injector mounted on the gas chromatograph It is also achieved by the method of distinguishing.

In addition, the above object of the present invention is also achieved by a method of distinguishing whole red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph, wherein the induction time is 7 minutes.

In addition, the object of the present invention is that the indole is any one or more of 1H-indole, 4-methyl indole and 1-ethyl indole, a healthy pepper and two bottles using a solvent-free solid sample injector mounted on a gas chromatograph It is also achieved by the method of distinguishing red pepper.

Details of the above objects and technical configurations and the effects thereof according to the present invention will be more clearly understood by the following detailed description with reference to the drawings showing preferred embodiments of the present invention. In addition, in the drawings, the length, thickness, etc. of layers and regions may be exaggerated for convenience. Like numbers refer to like elements throughout.

1 is a perspective view of a solvent-free solid sample injector used in the present invention.

Referring to FIG. 1, the solvent-free solid sample injector 100 includes a support 110, an injector 120, a glass capillary mill 130, an adjusting screw 140, and the like. As the solvent-free solid sample injector 100 can be used a keel gas chromatograph injector (Keele GC Injector), it is used by modifying to easily discharge the vaporized sample.

The injector 120 is a portion mounted to the injector port of the gas chromatograph, and the glass capillary in which the sample to be analyzed is sealed is inserted therein. In addition, a plurality of outlets 125 were formed in the injector 120 so as to be easily guided to the gas chromatograph column.

The glass capillary mill 130 serves to break the glass capillary in which the vaporized sample is sealed.

2A to 2C are schematic diagrams illustrating a method of analyzing aroma components of red pepper using a gas chromatograph equipped with a solvent-free solid sample injector used in the present invention.

Referring to Figures 2a to 2c will be described a method of analyzing the fragrance component of the pepper sample using the gas chromatograph 200 equipped with the solvent-free solid sample injector 100.

Referring to Figure 2a, the gas chromatograph 200 is a device for analyzing the fragrance components, the injector port 210, the heating means 220, the column 230 and the analysis means 240 is provided have.

At this time, the column 230 used HP-5MS (30mm length × 0.25mm i.d., 0.25㎛ film-thickness), the analysis means 240 used Hewlett Packard 6890N as GC / MSD.

The injector port 210 is where the injector 120 of the solventless solid sample injector 100 is mounted. The heating means 220 vaporizes a sample to be analyzed by heating the mounted injector 120. The column 230 is connected to the analysis means 240 so that the vaporized sample is led to the analysis means 240. In addition, the structure or operation principle of the gas chromatograph 200 will be omitted since it is obvious to those skilled in the art.

Then, the pepper to be analyzed is cut into a predetermined amount to prepare a pepper sample 160, and after sealing one end of the glass capillary 150, the prepared pepper sample 160 is injected, and then the glass capillary 150 Prepare by sealing the other end of the line.

At this time, the pepper sample 160 is used as it is without going through a separate solvent extraction process.

Subsequently, the injector 120 of the solvent-free solid sample injector 100 is mounted on the injector port 210, and the glass capillary 150 sealed with the red pepper sample 160 enters the injector 120. Loosen the glass capillary mill 130 so as to be able to.

Referring to FIG. 2B, the glass capillary 150 in which the red pepper sample 160 is sealed is inserted into the injector 120. Subsequently, the glass capillary mill 130 is inserted into the predetermined portion of the injector 120 through the adjusting screw 140.

Subsequently, the injector 120 is preheated by using the heating means 220 to vaporize the pepper sample 160 sealed in the glass capillary tube 150 inserted into the injector 120. In this case, the preheating temperature of the injector 120 may be adjusted by adjusting the power applied to the heating means 220.

Referring to FIG. 2C, the glass capillary 150 is pulverized so that the pepper sample 161 completely vaporized in the glass capillary 150 may be guided to the analyzing means 240 through the column 230. . At this time, the glass capillary tube mill 130 is pressed in the vertical direction to crush the glass capillary tube 150.

The vaporized pepper sample 161 is led to the column through an outlet 125 formed in the injector 120. In this case, a plurality of outlets 125 are formed in the injector 120 so that the vaporized pepper sample 161 may be led to the column 230. In addition, the time for directing the vaporized pepper sample 161 to the column 230 of the gas chromatograph may be appropriately adjusted.

The vaporized pepper sample 161 derived enters into the analysis means 240 and its fragrance component is analyzed.

In this case, the remaining molecules that are not vaporized may be prevented from being introduced into the column 230 by removing the solvent-free solid sample injector 100 after the analysis is completed, thereby preventing the column 230 from being contaminated.

Experimental Example 1

This experimental example analyzes the fragrance components of healthy red peppers and diseased red peppers infected with anthrax using the gas chromatograph 200 equipped with the solvent-free solid sample injector 100 of the present invention described with reference to FIGS. 2A to 2C. It was.

First, prepare healthy red peppers purchased from farmhouses and two hot peppers with anthrax.

Then, the healthy peppers and two bottled peppers are cut into a predetermined amount, respectively, to prepare a certain amount of healthy pepper samples and two bottled pepper samples.

The fragrance components of the whole red pepper sample and the two red pepper samples were analyzed using the gas chromatograph 200 equipped with the solvent-free solid sample injector 100 described with reference to FIGS. 2A to 2C.

The results are shown in Table 1 below.

No. Compounds RT RI Area (%) Whole red pepper Two peppers One 3-Methylbutanal 2.07 <800 2.69 6.43 2 Acetic acid 2.37 <800 32.04 2.59 3 2-Propanone 2.42 <800 4.15 5.35 4 Pyrazine 2.84 <800 - 7.56 5 1H-Pyrrole 3.16 <800 - 3.73 6 1,2-Ethanediamine 3.70 <800 - 2.89 7 3-Amino-1-propanol 3.89 801.4 0.69 - 8 1,4-Dideuterio-2-methylbutane 4.02 806.9 - 1.28 9 Trimethylurea 4.19 814.2 0.81 4.3 10 2 [3H] -Furanone 4.37 821.1 - 1.26 11 Topotecan 4.43 823.6 - 1.18 12 2-Methylpyrazine 4.63 831.2 - 4.79 13 2-Furanmethanol 5.95 874.0 9.16 14.52 14 2,6-Dimethylpyrazin 7.55 920.6 - 2.16 15 Ethylpyrazine 7.65 923.8 - 1.41 16 Butyrolactone 7.84 929.6 4.3 3.34 17 2-Furancarboxaldehyde 9.43 973.7 2.32 - 18 2-Ethyl-6-methyl-pyrazine 10.62 1003.5 - 0.72 19 2-Ethyl-5-methyl-pyrazine 10.73 1006.9 - 0.49 20 2,3,5-Trimethylpyrazine 10.80 1009.5 - 1.96 21 Endo-2-methyltricyclo [4,10] decane 11.42 1029.5 - 0.85 22 2-Cyclopenten-1-one 11.83 1041.9 1.86 1.21 23 Benzeneacetaldehyde 12.15 1051.6 - 0.65 24 1- [1H-pyrrol-2-yl] -Ethanone 13.08 1078.0 - 0.97 25 2,5-Dimethyl-4-hydroxy-3 [2H] -furanone 13.32 1084.5 3.64 1.68 26 Cyclobutanol 13.98 1102.6 3.39 3.72 27 1-Propanol 14.23 1111.3 - 0.58 28 1-Guanidinosuccinimide 14.31 1114 - 1.04 29 4H-Pyran-4-one 14.61 1124.4 0.99 - 30 3-Ethyl-2-hydroxy-2-Cyclopenten-1-one 14.79 1130.5 - 0.38 31 Erythro-1,2,4-trimethylpnet-4-en-1-ol 15.27 1146.5 - 0.64 32 β-D ₂ -γ-Picoline 15.36 1149.3 - 0.96 33 4-Pyridinol 15.56 1155.8 - 3.11 34 3-Hydroxypyridine 15.9 1166.5 33.49 9.93 35 Decanal 16.55 1186.4 0.47 0.24 36 6-Methyl-3-pyridinol 17.01 1200 - 1.42 37 5-Ethyldihydro-2 [3H] -furanone 17.26 1209.3 - 1.41 38 4-Pyridinamine 19.63 1292.4 - 0.96 39 1H-Indole 19.95 1303.4 - 0.62 40 1,3-Benzenediamine 21.86 1376.2 - 0.92 41 4-Methylindole 22.39 1395.4 - 0.61 42 1-Ethylindole 24.79 1491.6 - 1.58 43 Acetamide 25.59 1525.5 - 0.57 Total 100 100

In this case, the compound was identified by a GC / MS library search program, the RT (Retention Time) is the duration time in HP-5MS of GC-MS, the RI (Retention indices) is HP-5MS of GC-MS Calculated duration in, and Area (%) represents the relative peak area calculated from the GC-MS total ion chromatograph.

As shown in the data of Table 1, the healthy red peppers and two hot peppers were detected with 14 kinds of fragrance components and 40 kinds of fragrance components, respectively.

The whole red pepper was detected with 3-hydroxypyridine (33.49% of the total area) and acetic acid (32.04%) as main components, and 2-furanmethanol and minor A large amount of butyrolactone was detected, 2-furanmethanol (14.52%) and 3-hydroxypyridin (9.93%) were the main components of the diseased pepper, and pyrazine and 3-methylbutal (3- Methylbutanal was also detected in large quantities.

Many heterocyclic compounds, such as pyrazine, were detected in the diseased peppers, which may have a significant effect on the aroma of the diseased peppers.

In addition, indole compounds (1H-Indole, 4-Methylindole and 1-Ethylindole) were detected in two peppers, but these compounds were not detected in whole peppers. .

Therefore, the difference between the healthy pepper and the diseased pepper from the above results is that the indole, i.e., 1H-indole, 4-methyl indole and 1-ethylindole is detected whether the indole is an indicator material for determining the presence of anthrax in the pepper Able to know.

That is, when the indole is detected, the anthrax is infected with two diseased peppers, and when the indole is not detected, it can be seen that the anthracnose is an unhealthy whole pepper.

Experimental Example 2

In this experimental example, the fragrance components of the whole red pepper sample and the two red pepper samples were analyzed using the gas chromatograph 200 equipped with the solvent-free solid sample injector 100 of the present invention described with reference to FIGS. 2A to 2C. Experimental example for setting the optimal condition for

Among the fragrance components analyzed by the solvent-free solid sample injector (see Table 1 above), 1H-pyrrole, 3-hydroxypyridine, and 1H-indole were selected to analyze the analytical efficiency under various conditions. .

At this time, the analysis means and the column used in the present experimental example used the same as described with reference to Figs. The temperature program was maintained at 50 ° C. for 5 minutes, and then raised to 280 ° C. at a flow rate of 5 ° C./min, and held for 5 minutes.

In addition, the carrier gas (Carrier gas) used helium, the detector temperature was 300 ℃.

Looking at the preparation process and results of the experiment to analyze the flavor components of the pepper samples are as follows.

First, prepare pepper, cut the pepper to have a weight of 1mg to prepare a pepper sample.

Then, the pepper sample is placed in a glass capillary sealed at one end, and the other end is sealed and injected into the gas chromatograph using a solvent-free solid sample injector.

At this time, the preheating temperature of the injector to volatilize the pepper sample sealed in the glass capillary was 200 ℃, 250 ℃ and 300 ℃ experiments, respectively, the preheating time of the injector was 3, 5, 7 and 10 minutes respectively The evaporated pepper samples were tested for 0, 3, 5, 7 and 10 minutes, respectively.

3 is a graph showing the effect of the preheating temperature of the injector to detect the flavor component of the pepper.

Referring to Figure 3, it can be seen that the analysis efficiency according to the preheating temperature of the injector from the relationship between the preheating temperature of the injector and the detector response, the 1H-pyrrole and 3-hydroxypyridine is the highest analysis efficiency at 250 ℃ 1H-indole showed the highest analysis efficiency at 300 ° C. At this time, when considering that the 1H-indole is a component appearing only in the diseased pepper, 300 ℃ should be the optimal preheating time, but when analyzed at 300 ℃ the baseline of the chromatogram is not stable, the optimal preheating temperature of the injector is 250 It is preferable that it is ° C.

Figure 4 is a graph showing the effect of the preheating time of the injector to detect the flavor component of the pepper.

Referring to Figure 4, it can be seen that the analysis efficiency according to the preheating time of the injector from the relationship between the preheating time of the injector and the detector response, the 1H-pyrrole and 1H-indole were detected almost all the time, but the 3-hydr In the case of oxypyridine, 7 minutes had the highest analysis efficiency. Therefore, the optimum preheating time of the injector is preferably 7 minutes.

5 is a graph showing the effect of the time to induce the vaporized pepper sample to the column of the gas chromatograph on the flavor component detection of pepper.

Referring to Figure 5, all three components of 1H-pyrrole, 3-hydroxypyridine and 1H-indole shows the best analytical efficiency when the vaporized pepper samples are induced for 7 minutes into the column of the gas chromatograph It can be seen that.

Therefore, the method of distinguishing whole red peppers and two red peppers using the solventless solid sample injector mounted on the gas chromatograph of the present invention is to mount the injector of the solventless solid sample injector to the injector port provided in the gas chromatograph, After inserting the glass capillary sealed with the pepper sample into the injector of the solvent-free solid sample injector, the injector is preheated to vaporize the capsicum sample sealed therein, and the glass capillary is broken to vaporize the pepper sample. After analyzing the fragrance component of the vaporized pepper sample by inducing the column of the gas chromatograph, judging whether or not indole is contained in the fragrance component of the red pepper sample, if the indole is contained, If it does not contain indoles, it is judged as healthy red pepper Infected can be detected anthrax in early May to be able to distinguish between healthy or diseased peppers and chili will not be able to reduce the economic losses to the initial control.

In addition, the optimum conditions for analyzing the fragrance components of the whole red pepper sample and the two bottle pepper sample is the preheating temperature of the injector is 250 ℃, the preheating time of the injector is 7 minutes, the vaporized pepper samples of the gas chromatograph The time to lead to the column is preferably 7 minutes.

Although the present invention has been shown and described with reference to preferred embodiments as described above, it is not limited to the above-described embodiments and those skilled in the art without departing from the spirit of the present invention. Various changes and modifications will be possible.

Therefore, the method of distinguishing whole red peppers and two red peppers using the solvent-free solid sample injector mounted on the gas chromatograph of the present invention analyzes the fragrance component of the red pepper sample using the gas chromatograph injector of the solvent-free solid sample injector. If it is included, judging from the diseased peppers, if it does not contain indoles, there is an effect of presenting a method for distinguishing between the diseased peppers and healthy peppers judged as healthy peppers.

In addition, the pepper sample is not only simple and fast through the solvent extraction process, but also has the effect of accurately distinguishing healthy peppers and two diseased peppers by accurately analyzing the fragrance components without loss of fragrance components.

In addition, by analyzing the fragrance component of the pepper sample to provide a method for accurately distinguishing anthrax early pepper has the effect of early control of pepper anthrax.

Claims (9)

Mounting an injector of a solvent-free solid sample injector to an injector port provided in a gas chromatograph;  Inserting a glass capillary sealed with a predetermined amount of pepper samples into the injector of the solvent-free solid sample injector;  Preheating the injector to vaporize the pepper sample sealed therein;  Breaking the glass capillary to guide the vaporized pepper sample to a column of the gas chromatograph;  Analyzing the fragrance components of the evaporated chili pepper sample; And  Judging whether indoles are contained in the fragrance component of the red pepper sample, judging as two red peppers if indoles are included, and judging as whole red peppers if indoles are not included. Distinguishing method between healthy red peppers and two red peppers using a solvent-free solid sample injector mounted on a gas chromatograph. The method of claim 1, The injector of the solvent-free solid sample injector is a healthy pepper using a solvent-free solid sample injector mounted on the gas chromatograph, characterized in that it has a plurality of holes to guide the vaporized pepper sample to the column of the gas chromatograph. To distinguish between and diseased peppers. The method according to claim 1 or 2, The preheating temperature of the injector is 200 to 300 ℃ characterized in that the dry pepper and the bottle pepper using a solvent-free solid sample injector mounted on a gas chromatograph. The method of claim 3, wherein The preheating temperature is 250 ℃ method for distinguishing between healthy red peppers and two peppers using a solvent-free solid sample injector mounted on a gas chromatograph. The method according to claim 1 or 2, The preheating time of the injector is 3 to 10 minutes, characterized in that the distinction between the healthy pepper and the bottle pepper using a solvent-free solid sample injector mounted on the gas chromatograph. The method of claim 5, wherein The preheating time is 7 minutes, characterized in that the distinction between the healthy pepper and the bottle pepper using a solvent-free solid sample injector mounted on the gas chromatograph. The method according to claim 1 or 2, The time for inducing the evaporated sample to the column of the gas chromatograph is 10 minutes or less, characterized in that the distinction between the healthy pepper and the diseased pepper using a solvent-free solid sample injector mounted on the gas chromatograph. The method of claim 7, wherein The induction time is 7 minutes, characterized in that the distinction between healthy peppers and two peppers using a solvent-free solid sample injector mounted on a gas chromatograph. The method according to claim 1 or 2, The indole is one or more of 1H-indole, 4-methyl indole and 1-ethyl indole, characterized in that the distinction between the healthy pepper and the diseased pepper using a solvent-free solid sample injector mounted on the gas chromatograph.

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