KR20170110795A - Analytical method to detect α-dicarbonyl compounds, including glyoxal, methylglyoxal and diacetyl - Google Patents

Abstract

The present invention relates to a method for determining whether an alpha-dicarbonyl compound is contained in a sample.
More specifically, according to the present invention, the presence or absence of an alpha-dicarbonyl compound in a sample is detected with high sensitivity using a dispersive liquid-liquid microextraction (DLLME) and gas chromatography-mass spectrometry Thus, it is possible to accurately and conveniently quantitatively determine whether or not the alpha-dicarbonyl compound having toxicity in the sample is contained.

Description

[0002] Analytical methods to detect α-dicarbonyl compounds, including glyoxal, methylglyoxal and diacetyl, which include α-dicarbonyl compounds such as glyoxal, methylglyoxal or diacetyl,

The present invention relates to a method for qualitatively and quantitatively analyzing the presence of an? -Dicarbonyl compound in a sample, and more specifically, to a method for qualitatively and quantitatively analyzing the presence or absence of a toxic? -Dicarbonyl compound such as glyoxal, methylglyoxal or diacetyl Dicarbonyl compounds in oil-soluble products such as water-soluble products and sesame oil and perilla oil, such as red ginseng products, and quantitatively analyzing the contents thereof.

Alpha-dicarbonyl compounds are produced by oxidation or myral reactions of sugars or lipids by heat treatment. Typical examples are glyoxal, methylglyoxal, and diacetyl, and their toxicity There have been many reports.

Glyoxal is an organic compound of the formula C ₂ H ₂ O ₂ , which is a yellow liquid that evaporates into a green gas. Glyoxal is an inflammatory compound that is produced when cooking oils and fats are heated to high temperatures, usually through contact with the respiratory or skin. There is a possibility of inducing irritation of inhaled glyoxal (Prager, JC Environmental Contaminant Reference Databook Volume 1. New York, NY: Van Nostrand Reinhold, 1995, p. 729) (New York, NY: Van Nostrand Reinhold, 1995, p. 729). In addition, it has been reported that the causes of dermatitis and mutagenesis have been reported. In addition, glyoxal is found in the blood of diabetic patients and is found in high concentrations in patients with renal failure.

Methylglyoxal is an organic compound of the formula C ₃ H ₄ O ₂ , which is a yellow liquid with a slight viscosity giving a pungent odor. It has been reported that rats exposed to methylglyoxal are abnormally increased in body weight and develop early insulin resistance and type 2 diabetes. It is predicted that methylglyoxal will have a deleterious effect on the body's important end-glycated product receptor activities such as AGER1, which causes a significant reduction of the body's protective mechanisms such as SIRT1 and also protects SIRT1 and inhibits insulin resistance. In addition, methylglyoxal is also associated with arterial atherogenesis, and low density lipoprotein damage by glycation of methylglyoxal has been reported to cause a 4-fold increase in atherogenesis in diabetes (Rabbani N et al., (May 26, 2011). "Glycation of LDL by methylglyoxal increases arterial atherogenicity." A possible contributor to increased risk of cardiovascular disease in diabetes. "Diabetes 60 (7): 1973-80.)

Diacetyl is a colorless liquid with a strong chlorine-like odor of the formula C ₄ H ₆ O ₂ , which can be absorbed into the body by the respiratory or feeding. Causes coughing, vomiting and headache when absorbed into the respiratory tract. Causes redness on skin contact and may cause redness on contact with eyes. In workplaces dealing with diacetyl, young and healthy non-smoking workers have been diagnosed with bronchiolitis obliterans, and rarely lung diseases have been reported. In addition, there have been reports that diacetyl may exacerbate beta-amyloid accumulation, which is associated with Alzheimer's (Swati S et al., (2012). "The Butter Flavorant, Diacetyl, Exacerbates β- Amyloid Cytotoxicity" Chemical Research in Toxicology). Therefore, the California Council of the United States is proposing to ban diacetyl, and the European Food Safety Authority (EFSA) is reviewing the toxicity of diacetyl.

Recently, alpha-dicarbonyl compounds have been detected in processed products such as some butter, coffee, honey, yogurt, carbonated beverages, and they are toxic substances. The alpha-carbonyl compound has been found in the present invention to be contained in red ginseng products, one of health functional foods. This predicts that lipids would have been produced by oxidation during processing, but the exact path was not revealed. In the case of diacetyl, NIOSH (National Institute of Occupational Safety and Health) has its limit tolerance (<20 μg / g), but it is limited , So far the safe tolerance of toxicity of alpha-dicarbonyl compounds including diacetyl has not been established in Korea. Therefore, it is necessary to accurately and conveniently detect the presence of the alpha-dicarbonyl compound in the product before the tolerance threshold is settled, and it is necessary to develop a quantitative analysis method thereof.

A. Leonardis et al. Describe a method for analyzing the diacetyl content of dairy products without solvent purification using solvent extracts, but the method is limited to dairy products containing high amounts of diacetyl . In addition, Yong Chen et al. Have developed a method for analyzing the content of diacetyl that is generated from volatile butter and popcorn using Solid Phase Microextraction (SPME), but the SPME method has a disadvantage of requiring expensive apparatus. In addition, Takayuki Shibamoto et al. Have developed a method for the analysis of diacetyl produced by the heating of lipid of food using headspace sampling method, but the use of toxic solvent is inevitable for headspace sampling technique.

Meanwhile, according to the Food Additives Code of the Food and Drug Administration at Korea Food and Drug Administration, diacetyl analysis of a popcorn product for a microwave oven is being proposed as another method for qualitative analysis and quantitative analysis. Solid phase microextraction (SPME) , And the quantitative method is described as a solvent extraction using Simultaneous Distillation Extraction (SDE) apparatus.

Accordingly, an object of the present invention is to provide a method for accurately and conveniently detecting the presence of an alpha-dicarbonyl compound in a sample in a short time.

More specifically, the present invention provides a method for detecting the presence of an alpha-dicarbonyl compound in a sample using a dispersive liquid-liquid microextraction (DLLME) and gas chromatography-mass spectrometry. Through these, it is possible to provide a method capable of quantitative analysis of alpha-dicarbonyl compounds.

In order to solve the above-mentioned problems, the present invention provides a method for determining whether an alpha-dicarbonyl compound is contained in a sample:

(S1) adding o-phenylenediamine dihydrochloride (OPD) to a sample and reacting;

(S2) adding an extraction solvent and a dispersion solvent according to a dispersive liquid-liquid microextraction (DLLME) method and centrifuging; And

(S3) analyzing the alpha-dicarbonyl compound in the extraction solution with a gas chromatograph-mass spectrometer,

Chloroform in the extraction solvent, and methanol in the dispersion solvent.

According to a preferred embodiment of the present invention, the sample may be red ginseng product, perilla oil product or sesame oil product.

According to a preferred embodiment of the present invention, the step (S1) further comprises the step of adding an internal standard substance to the sample, wherein the internal standard substance is 1-methylpyrazole (1-MP) Can be used.

According to one preferred embodiment of the present invention, the alpha-dicarbonyl compound may be glyoxal, methylglyoxal or diacetyl.

According to a preferred embodiment of the present invention, the ion (m / z) of quinoxaline is 130, the ion (m / z) of methylquinoxaline is 144, or 2,3-dimethyl The ion (ion, m / z) of quinoxaline may be 158.

According to a preferred embodiment of the present invention, the volume of the dispersing solvent relative to the volume of the extracting solvent may be 1: 1-3.

Hereinafter, the present invention will be described in detail.

The present invention relates to a method for determining the presence of an alpha-dicarbonyl compound in a sample, comprising the steps of:

(S1) adding o-phenylenediamine dihydrochloride (OPD) to a sample and reacting;

(S2) adding an extraction solvent and a dispersion solvent according to a dispersive liquid-liquid microextraction (DLLME) method and centrifuging; And

(S3) analyzing the alpha-dicarbonyl compound in the extraction solution with a gas chromatograph-mass spectrometer,

Chloroform in the extraction solvent, and methanol in the dispersion solvent.

The present inventors have found that when using a dispersive liquid-liquid microextraction (DLLME) and gas chromatography-mass spectrometry to determine the presence of an alpha-dicarbonyl compound in a sample, -Dicarbonyl compound can be accurately and conveniently detected with high efficiency, and the present invention has been completed. Particularly, o-phenylenediamine dihydrochloride (OPD) is used as the derivative conversion reagent, preferably chloroform as the extraction solvent and methanol as the dispersion solvent. According to the present invention, it is possible to quantitatively analyze not only qualitative analysis but also to detect toxic alpha-dicarbonyl compounds with high efficiency and high accuracy, in particular, within a recovery rate of 90 to 120%. In addition, when the internal standard substance is added before extraction, the convenience is enhanced and high-efficiency detection is possible.

Hereinafter, the method according to the present invention will be described step by step:

The present invention relates to a method for determining the incorporation of alpha-dicarbonyl compounds in a sample.

In the present invention, it is sufficient that the 'sample' is suspected of containing the alpha-dicarbonyl compound, and whether or not the alpha-dicarbonyl compound is actually included. The sample includes red ginseng product, perilla oil product or sesame oil product. The sample may be pretreated to be applicable to the dispersion liquid-liquid microextraction method, for example, liquefied, and is not limited to an aqueous solution. According to a specific embodiment of the present invention, a sample can be prepared, for example, by adding a phosphate buffer after dissolving the red ginseng product. In addition, various additives (for example, antifoaming agents) that can be added to the samples according to the dispersed liquid-liquid microextraction method can be added to the samples.

In the present invention, 'red ginseng product' refers to a product in which the main ingredient of the sample is recognized as red ginseng. If red ginseng is described as a raw material, it is regarded as a red ginseng product according to the present invention. The red ginseng product is not limited to a specific formulation such as a powder, a lump, a paste or a liquid, and it may suffice to liquefy it before extraction. Preferably, the state of the sample before extraction may be a liquid phase. The above products are not limited to those commercialized and distributed, and may be included in the case of articles containing red ginseng as a raw material.

In the present invention, 'perilla oil product' or 'sesame oil product' refers to a product in which the main component of the sample is perceived to be perilla oil or sesame oil. If perilla oil or sesame oil is described in the raw material name, perilla oil product or sesame oil I regard it as a product. The state of the sample before extraction may be liquid. The above products are not limited to those that are commercialized and distributed, and may be included in all cases of products made from perilla oil or sesame oil.

In the present invention, 'ginseng (hongsam, red ginseng)' refers to steamed ginseng Panax ginseng CA Meyer (Araliaceae), and the ginseng that has undergone steaming under the same scientific name as the above, It is considered to be included in red ginseng, and does not discriminate ginseng parts (for example, removal of bark, stem, root, etc.), habitat, cultivation or wild, harvest time. Preferably, red ginseng does not contain alpha-dicarbonyl compounds.

In the present invention, 'perilla oil' refers to the oil obtained by pressing the perilla after heating (preferably, roasting process), and perilla (Perilla frutescens var. Japonica Hara) It is an annual plant, and its origin is in the highlands of India and central and southern China. The perilla in the present invention is not limited to a particular one in terms of mountain area, harvest season, and breed. In addition, it is not limited to a specific part of the perilla, as long as perilla oil can be obtained, but it is preferably seed. Preferably, the natural perilla, for example, which is not subjected to a heating step, contains no alpha-dicarbonyl compound.

In the present invention, 'sesame oil' refers to the oil obtained by pressing the sesame after heating (preferably roasting) and then pressing the 'Sesamum indicum', which is an annual plant of sesame seeds of the dicotyledonous plant, Is an annual herbaceous plant widely distributed in Korea, East Asia, North America and Africa. The sesame seeds of the present invention are not limited to those specific to the origin, harvest date, variety, and color. In addition, as long as sesame oil can be obtained, it is not limited to a specific part of sesame, but it is preferably seed. Preferably, the natural sesame which has not undergone a heating step, for example, contains no alpha-dicarbonyl compound.

In the present invention, "glyoxal, methylglyoxal, diacetyl" in the alpha-dicarbonyl compound refers to a compound of the following formula (1)

[Chemical Formula 1]

Glyoxal: R ₁ = R ₂ = H

Methylglyoxal: R ₁ = CH ₃ , R ₂ = H

Diacetyl: R ₁ = R ₂ = CH ₃

(S1), adding o-phenylenediamine dihydrochloride (OPD) to the sample and reacting.

In the present invention, o-phenylenediamine dihydrochloride (OPD) is used as a derivative conversion reagent.

Since the analyte according to the present invention has polarity, it can be adsorbed on a gas chromatography column or decomposed in an injector to deteriorate the reproducibility of analysis. In order to solve this problem, a polar material is converted into a non- . In the present invention, various compounds for converting an alpha-dicarbonyl compound to a non-polar material may be used, but most preferably, o-phenylenediamine dihydrochloride (OPD) may be used (FIG. 6). When o-phenylenediamine dihydrochloride is added to the alpha-dicarbonyl compound, it is converted to a quinoxaline derivative. The quinoxaline derivative is represented by the following formula (2). In the present invention, "quinoxaline, 2-methylquinoxaline, 2,3-dimethylquinoxaline" among the quinoxaline derivatives refers to a compound represented by the following formula (2).

(2)

Quinoxaline: R ₁ = R ₂ = H

Utilizing _{2-methyl-quinoxaline: R 1 = CH 3, R} 2 = H

2,3-dimethylquinoxaline: R ₁ = R ₂ = CH ₃

For example, when o-phenylenediamine dihydrochloride is added, glyoxal is converted to quinoxaline, methylglyoxal is converted to 2-methylquinoxaline, diacetyl is converted to 2,3-dimethylquinoxaline (Fig. 5).

Preferably, the concentration of the o-phenylenediamine dihydrochloride is at least 5 g L ^-1 , more preferably at least 5 10 g L ^- may ^1st. Since the detection efficiency is excellent at 5 g L ^-1 or more and there is no difference in efficiency at 5 g L ^-1 and 10 g L ^-1 , it is most economical to use at a concentration in the range of 5 - 10 g L ^-1 ).

In addition, the reaction time with o-phenylenediamine dihydrochloride does not significantly affect the conversion efficiency.

(S2) a step of adding an extraction solvent and a dispersion solvent according to a dispersive liquid-liquid microextraction (DLLME) method and centrifuging.

In the present invention, 'Dispersive liquid-liquid microextraction (DLLME)' utilizes the principle of liquid-liquid extraction and the advantage of the miniaturization technology of solid trace extraction. The three-component solvent system, That is, a sample, an extraction solvent which is not dispersed in water, and a dispersion solvent which is dispersed in water and extraction solvent. When a mixture of the extraction solvent and the dispersion solvent is injected into the sample, a small droplet cloud forms and the principle of the sample is shifted to the extraction agent by the distribution law. The present invention is not limited to the known methods in the art to which the present invention belongs, unless specifically described in the present invention.

In the present invention, the 'extraction solvent' should have a higher density than water and have a high storage capacity for the substance to be extracted, that is, a quinoxaline derivative, and a solvent having a low solubility in water may be used. And chloroform at the end of the repeated test was the most efficient extraction possible (FIG. 1 and FIG. 12).

In the present invention, the 'dispersing solvent' may be a solvent which is excellent in the mixing property of the aqueous solution layer and the organic layer, weak in interfacial tension between the two layers, and capable of dispersing the substance to be extracted, that is, quinoxaline derivative, However, the inventors of the present invention confirmed that it is preferable to use methanol after repeated tests on various solvents because it is possible to extract the most efficiently (FIGS. 2 and 13).

Preferably, the volume of the dispersing solvent to the volume of the extraction solvent is 1: 1-15, more preferably 1: 1-3, most preferably 1: 1: 1-2 (Figs. 3 and 4), more preferably 1: 8-15, and most preferably 1: 9-l, for detecting alpha-dicarbonyl compounds contained in perilla oil or sesame oil products, 10 (Figs. 14 and 15). Excellent preconcentration ratio and extraction recovery rate can be shown in the ratio.

Preferably, in applying the dispersed liquid-liquid microextraction method according to the present invention, the step of adding the 'internal standard material' to the sample may further include the step of mixing the alpha-dicarbonyl compound and Compounds whose functional groups are similar and whose peaks do not overlap as a result of gas chromatography-mass spectrometry can be used, and preferably 1-methylpyrazole (1-MP) can be used as an internal standard substance. This minimizes the loss of alpha-dicarbonyl compounds in the sample and allows the quantitative analysis of alpha-dicarbonyl compounds.

In the present invention, the 'internal standard material' may be added to the sample before adding 'extraction solvent' or 'dispersion solvent', and the order of addition of the 'derivative conversion catalyst reagent' and 'internal standard material' And may be added at the same time. Preferably, an internal reference material may be added to the sample in step (S1).

Preferably, when applying the dispersion liquid-liquid microextraction method according to the present invention, various compounds known in the art can be selectively added and used. However, when a red ginseng product is used as a sample, have. The present inventors confirmed that the addition of sodium chloride did not affect the detection efficiency when the red ginseng product was used as a sample (FIG. 8).

In the present invention, the 'derivative conversion reagent', 'extraction solvent', 'dispersion solvent', 'internal standard substance', and other substances that can be added in step (S1) and / or step (S2) Shaking and centrifuging, and the process may be repeated one or more times.

According to a preferred embodiment of the present invention, other steps may not be added other than the step S2 before the analysis step S3. Preferably, a step of back extraction (which means a step of back extraction by adding an aqueous solution to an extraction solvent which is an organic solvent after the extraction step) may not be added.

(S3) analyzing the alpha-dicarbonyl compound in the extraction solution with a gas chromatograph-mass spectrometer.

Gas chromatography-mass spectrometry is used to analyze alpha-dicarbonyl compounds in the extraction solution. More specifically, the separated components are introduced into the mass spectrometer at different retention times using the difference in adsorptivity or partition coefficient between the stationary phase and the mobile phase of the capillary separation tube mounted on the gas chromatograph. The introduced sample is ionized by the ionization apparatus.

The ion (ion, m / z) of the quinoxaline derivative as the conversion compound of the alpha-dicarbonyl compound according to the present invention and 1-methylpyrazole as the internal standard are as follows:

The ion (ion, m / z) of 1-methylpyrazole is 82.

The ion (m / z) of quinoxaline is 130, the ion (m / z) of methylquinoxaline is 144, or the ion (m / z) of 2,3-dimethylquinoxaline is 158.

In yet another aspect, a kit is provided that is capable of applying the method according to the present invention. More specifically, the kit according to the present invention may include a tool capable of applying the dispersed liquid-liquid microextraction method and a guide describing a method of determining whether the alpha-dicarbonyl compound is contained in the sample .

Preferably, the kit according to the present invention may contain chloroform as the extraction solvent, methanol as the dispersion solvent, and o-phenylenediamine dihydrochloride as the derivative conversion reagent.

In addition, the above guidance describes how the method according to the present invention may be applied to those skilled in the art (e.g., the amount of extraction solvent, the amount of dispersion solvent, the amount of derivative conversion reagent, the quinoxaline derivative, Sol ions, etc.) may be described.

According to the present invention, the inclusion of alpha-dicarbonyl compounds in a sample can be detected with high sensitivity by using dispersive liquid-liquid microextraction (DLLME) and gas chromatography-mass spectrometry, This makes it possible to accurately and conveniently discriminate the presence of the alpha-dicarbonyl compound in the sample. Particularly, according to the present invention, excellent effects in terms of excellent linearity, accuracy, and detection limit can be obtained.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
1 is a graph showing extraction recovery (ER%) and preconcentration factor (FR%) according to three extraction solvents using DLLME, wherein (a) glyoxal, (b) methylglyoxal, (c) diacetyl .
(Extraction conditions: extraction solvent 100 μL, methanol 1 mL, OPD 10 g L ^-1 , reaction time 2 hours).
FIG. 2 is a graph showing the extraction recovery (ER%) and the preliminary concentration factor (FR%) according to the three dispersion solvents using the DLLME, wherein (a) glyoxal, (b) methylglyoxal, .
(Extraction conditions: chloroform 100 μL, dispersion solvent 1 mL, OPD 10 g L ^-1 , reaction time 2 hours).
Figure 3 is a graph showing the extraction recovery (ER%) and the preliminary concentration factor (FR%) according to the volume of the extraction solvent using the DLLME, wherein (a) glyoxal, (b) methylglyoxal, (c) diacetyl .
(Extraction conditions: extraction solvent chloroform, dispersion solvent 1 mL, OPD 10 g L ^-1 , reaction time 2 hours).
FIG. 4 is a graph showing extraction recovery (ER%) and preconcentration factor (FR%) according to the volume of the dispersing solvent using DLLME, wherein (a) glyoxal, (b) methylglyoxal, (c) diacetyl .
(Extraction conditions: chloroform 100 μL, dispersed solvent methanol, OPD 10 g L ^-1 , reaction time 2 hours).
FIG. 5 shows a reaction scheme in which an alpha-dicarbonyl compound and a derivative conversion reagent (OPD) react to form a quinoxaline derivative.
FIG. 6 is a graph showing the extraction recovery (ER%) and the preliminary concentration factor (FR%) according to the concentration of the derivative conversion reagent (OPD) using the DLLME, wherein (a) glyoxal, (b) methylglyoxal, c) corresponds to diacetyl.
(Extraction conditions: chloroform 100 μL, methanol 1 mL, reaction time 2 hours).
FIG. 7 is a graph showing the extraction recovery (ER%) and the preliminary concentration factor (FR%) according to the reaction time of the derivative conversion reagent (OPD) using the DLLME, wherein (a) glyoxal, (b) methylglyoxal, (c) diacetyl.
(Extraction conditions: chloroform 100 μL, methanol 1 mL, OPD 5 g L ^-1 ).
FIG. 8 is a graph showing extraction recovery (ER%) and preconcentration factor (FR%) according to the concentration of NaCl solution using DLLME, .
(Extraction conditions: chloroform 100 μL, methanol 200 μL, OPD 5 g L ^-1 , reaction time 1 hour).
Figure 9 shows the total ion chromatogram obtained from standard solutions of quinoxaline derivatives from alpha-dicarbonyl compounds (33.3 [mu] g L &lt; ^-1 &gt;) and 1-methylpyrazole (internal standard) using GC-MS.
Glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), 1-methylpyrazole (1-MP)
Fig. 10 shows the total ion chromatogram of alpha-dicarbonyl-derived quinoxaline derivatives obtained from red ginseng products (concentrate) using GC-MS.
Glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), 1-methylpyrazole (1-MP)
Figure 11 shows the total ion chromatogram of alpha-dicarbonyl-derived quinoxaline derivatives obtained from red ginseng products (beverages) using GC-MS.
Glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), 1-methylpyrazole (1-MP)
12 is a graph showing the peak area (%) according to the three extraction solvents using DLLME, and is a graph showing the peak areas (%) of quinoxaline (Q), 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline , 3-DMQ).
(Concentration of each compound, 250 ng mL ^-1 ; OPD, 0.6 mg; induction time, 2 hours; dispersion solvent, methanol; dispersion solvent volume, 0.2 mL; 0.5 M sodium phosphate buffer ))
13 is a graph showing the peak area (%) according to three kinds of dispersion solvent using DLLME, and is a graph showing the peak areas (%) of quinoxaline (Q), 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline , 3-DMQ).
(Extraction conditions: water, 3 mL; concentration of each compound, 250 ng mL ^-1 ; OPD, 0.5 mg; induction time, 2 hours; extraction solvent, chloroform; extraction solvent volume, 100 μL; 0.5 M sodium phosphate buffer ))
14 is a graph showing the peak area (%) according to the volume of the extraction solvent using the DLLME. As shown in FIG. 14, quinoxaline (Q), 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline , 3-DMQ).
(Extraction conditions: aqueous phase, 3 mL; concentration of each compound, 250 ng mL ^-1 ; OPD, 0.5 mg; induction time, 2 hours; dispersion solvent, methanol; dispersion solvent volume, 0.2 mL; extraction solvent, chloroform; Phosphate buffer (pH 7.6))
FIG. 15 is a graph showing the peak area (%) according to the volume of the dispersion solvent using the DLLME. As shown in FIG. 15, quinoxaline Q, 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline , 3-DMQ).
(Extraction conditions: aqueous phase, 3 mL; concentration of each compound, 250 ng mL ^-1 ; OPD, 0.5 mg; induction time, 2 hours; dispersion solvent, methanol; extraction solvent, chloroform; extraction solvent volume, 100 μL; 0.5 M sodium Phosphate buffer (pH 7.6))
FIG. 16 is a graph showing the peak area (%) according to the mass of the derivative conversion reagent (OPD) using DLLME. As shown in FIG. 16, quinoxaline Q, 2-methylquinoxaline 2-MQ, Corresponds to quinoxaline (2,3-DMQ).
(Extraction conditions: water, 3 mL; concentration of each compound, 250 ng mL ^-1 ; induction time, 2 hours; dispersion solvent, methanol; dispersion solvent volume, 0.2 mL; extraction solvent, chloroform; extraction solvent volume, 100 μL; 0.5 M sodium phosphate buffer (pH 7.6))
17 is a graph showing the peak area (%) according to the derivative conversion time using DLLME, which corresponds to quinoxaline (Q).
(Extraction conditions: water, 3 mL; concentration of each compound, 250 ng mL ^-1 ; OPD, 1000 μg; dispersion solvent, methanol; dispersion solvent volume, 0.2 mL; extraction solvent, chloroform; extraction solvent volume, 100 μL; 0.5 M Sodium phosphate buffer (pH 7.6))
Figure 18 shows a chromatogram of an alpha-dicarbonyl compound standard solution (20 ng mL ^{- 1} of each compound).
Quinoxaline (Q), 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline (2,3-DMQ)
19 shows the chromatogram of alpha-dicarbonyl compound in sesame oil (sample 1).
Quinoxaline (Q), 2-methylquinoxaline (2-MQ), 2,3-dimethylquinoxaline (2,3-DMQ)
Figure 20 shows the effect of roasting temperature and time on the generation of alpha-dicarbonyl compounds in sesame oil (left) and perilla oil (right).

Hereinafter, production examples and examples will be described in detail to facilitate understanding of the present invention. However, the production examples and examples according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following production examples and examples. The production examples and embodiments of the present invention are provided to enable those skilled in the art to more fully understand the present invention.

[In the red ginseng products, α- Dicarbonyl  Detection of compound]

1. Experimental material

(1) Red ginseng concentrate and red ginseng beverage sample

Red ginseng concentrate and red ginseng beverage samples were purchased at the market mart. Samples were stored at room temperature or in refrigeration according to the storage method indicated on the product.

(2) Reagents and reference materials

Glyoxal (ethanedial), 2-methylglyoxal (2-oxopropanol), diacetyl (2,3-butanedione, 2, 3-butanedione, 1-methylpyrazole (1-MP), o-phenylenediamine dihydrochloride (OPD), quinoxaline, 2-methylquinoxaline 2-methylquinoxaline, 2,3-dimethylquinoxaline, sodium phosphate dibasic and antifoam Y-30 emulsion were prepared according to Sigma Aldrich (St. Louis, MO, USA). HPLC grade tetrachlorethylene and chlorobenzene were purchased from Junsei (Tokyo, Japan). HPLC grade acetonitrile (ACN), methanol, chloroform, acetone, Milli-Q water purification system (Millipore, Billerica, MA, USA) ).

(1 g · L ^{- 1} ) and 1-MP (internal standard) working solution of glyoxal, methylglyoxal, diacetyl (reference material) working solution (10 mg · L ^{- 1} ) were prepared in methanol and distilled water (DW), respectively. A working standard solution of glyoxal, methylglyoxal and diacetyl was prepared by diluting a stock solution of glyoxal, methylglyoxal and diacetyl with distilled water (DW) (3.3, 16.7, 33.3, 166.7, 333.3 μg L ^{- 1} ).

2. Experimental Method

(1) Red ginseng concentrate and red ginseng beverage alpha - Dicarbonyl  The compound (? Dicarbonyl  Compounds for analysis Dispersive  Liquid-Liquid Microextraction  ( DLLME ) Way

Red ginseng concentrate was prepared by adding 1 mL of 10 g L ^-1 antifoaming agent Y-30 to 5 g, followed by 100 mL of second distilled water. After that, sonication was carried out at room temperature for 5 minutes to completely dissolve the red ginseng concentrate.

1 mL of red ginseng concentrate solution, 1 mL of red ginseng beverage, 1 mL of sodium phosphate buffer solution (0.5 M, pH 7.6), 1 mL of distilled water (total 3 mL) in a 10 mL conical glass tube, Respectively. 100 μL of 1-MP internal standard solution of 10 mg L ^{- 1} and 100 μL of OPD (derivative conversion reagent) of 5 g L ^{- 1} were added thereto, and the mixture was stirred for 1 hour with a twister using a spin bar Lt; / RTI &gt; Then, 200 μL of methanol (dispersing solvent) and 100 μL of chloroform (extraction solvent) mixed solution were rapidly injected using a glass syringe. Then, it was shaken for 2 seconds. The solution was centrifuged at 2,500 rpm for 3 minutes, and the sediment phase chloroform extract solution (about 50-70 μL) was taken using a glass syringe and transferred to a 250 μL insert vial. Respectively. This method was repeated three times.

(2) Red-ginseng extract and red- Dicarbonyl  Validation for compound analysis

The relative calibration curve was obtained by converting and extracting the derivatives using the same method as the sample using glyoxal, methylglyoxal, and diacetyl storage solutions. Recovery rate was measured by adding glyoxal, methylglyoxal, diacetyl (standard substance) 16.7, 33.3, and 166.7 μg L ^-1 in red ginseng concentrate and red ginseng beverage. For GC-MS instrumental analysis of glyoxal, methylglyoxal and diacetyl, the linearity of 3.3-333.3 mg L ^-1 standard solution was confirmed. The test solution of glyoxal, methylglyoxal, and diacetyl standard substances obtained after extraction was subjected to 6 repeated cycles of inter-day (3 days per day, 2 days) and intra-day (6 days per day) Respectively.

(3) Gas chromatography-mass spectrometry (GC-MS)

For the instrumental analysis, Agilent 7890 gas chromatograph and 5975c mass selective detector (Agilent Technologies, Palo Alto, Calif.) Were used. The extracted materials were analyzed using a DB-WAX column (30 m × 0.25 mm id, 0.25 μm film thickness, Agilent Technologies ). The injector temperature for the sample injection was 200 ° C, the helium gas flow rate was 1.0 mL min ^-1 , and the splitless mode. Oven programming and is again raised to then at 50 ℃ 2 minutes, the increase after three minutes at a speed of 10 min ^-1 for ℃ to 150 ℃ maintained for, 200 ℃ for 12 minutes at a speed of 10 min ^-1 ℃ held for twenty minutes Respectively. MS transfer line, MSD source and quadrupole temperature were set at 225, 230 and 150 ℃, respectively. Mass spectrum was recorded in SIM (Selected ion monitoring) mode. 1-MP is m / z 82, 81, 54, quinoxaline is from 15 minutes to m / z 130, 103, 76, 2-methylquinoxaline is from 17 minutes to m / z 144, 117, , 3-dimethylquinoxaline was monitored at m / z 158, 117, 76 from 18.4 minutes. m / z 82, 130, 144, and 158 were used as target ions for quantitative analysis of 1-MP, quinoxaline, 2-methylquinoxaline and 2,3-dimethylquinoxaline, respectively. The qualitative analysis was performed comparing with the ratio of ions in the standard material.

3. Experimental Results

(One) DLLME  Test extraction condition

Several variables affecting the extraction efficiency of the DLLME extraction method were changed. The optimized parameters are the extraction solvent, the type and volume of the dispersive solvent, and the NaCl concentration. The concentration of OPD used as a derivatizing agent, the preconcentration factor (PF) and extraction recovery (ER%) for the reaction time were calculated to maximize the conversion efficiency of the derivatives. Respectively. PF and ER are obtained as follows.

PF = Csed / C ₀

ER (%) = (Csed × Vsed / C ₀ × Vaq) × 100

        = PF x (Vsed / Vaq) x 100

The Csed is the concentration of the analyte in the sedimented phase obtained from the absolute calibration curve in the range of 0.05-5 mg L ^-1 and C ₀ is the initial concentration of the analyte in the aqueous phase. Vsed and Vaq refer to the sedimented phase and the volume of the aqueous sample in the aqueous solution, respectively.

(2) Selection of extraction solvent

For the selection of the extraction solvent, extraction efficiencies were compared using three extraction solvents (chloroform (CHCl ₃ ), chlorobenzene (C ₆ H ₅ Cl), tetrachlorethylene (CH ₂ Cl ₄ )). As a result, when chloroform was used, the preconcentration factor (PF) of glyoxal, methylglyoxal and diacetyl and the extraction recovery (ER%) were higher than those of chlorobenzene and tetrachlorethylene (FIG. 1) .

(3) Selection of dispersion solvent

For the selection of the dispersion solvent, extraction efficiencies were compared using three dispersion solvents (methanol (CH ₃ OH), acetonitrile (CH ₃ CN), acetone (C ₃ H ₆ O)). As a result, the preconcentration factor (PF) and recovery rate (ER%) of glyoxal, methylglyoxal, and diacetyl were the highest when methanol was used as compared with those using acetonitrile and acetone. Particularly, the pre-concentration factor (PF) of methanol showed a significantly higher difference than chlorobenzene and tetrachlorethylene (FIG. 2).

(4) Extraction solvent volume optimization

For the optimization of extraction solvent volumes, extraction efficiencies were determined for chloroform of 100, 150, 200, 250, and 300 μL. In the DLLME method performed in this study, the extraction solvent volume is an important factor affecting the preconcentration factor (PF). As the volume of the extraction solvent and the volume of the sedimented phase obtained after centrifugation increase, the value of the preconcentration factor (PF) decreases. Therefore, the extraction solvent volume optimization should obtain a volume of sedimented phase sufficient for proper analysis after centrifugation and a high value of the preconcentration factor (PF). In this study, the extraction yield (ER%) of glyoxal, methylglyoxal, and diacetyl was not significantly different for 100 μL of the extraction solvent, but the preconcentration factor (PF) was the highest at 100 μL of the extraction solvent. As a result, the volume of the extraction solvent chloroform was optimized to 100 μL (FIG. 3).

(5) Optimization of dispersion solvent volume

In order to optimize the volume of the dispersion solvent, extraction efficiencies of 200, 400, 600, 800 and 1000 μL of methanol were measured. The volume of the dispersed solvent has a direct influence on the formation of the water / dispersing solvent / extracting solvent, that is, the degree of dispersion of the extraction solvent in the aqueous solution. Depending on the volume of the dispersed solvent, the volume of the sedimented phase and the extraction recovery (ER%) value are different. As a result, the preconcentration factor (PF) in each volume did not show a large difference, but the volume of the sedimented phase tended to increase as the volume of the dispersion solvent became smaller, ER%). Thus, the volume of the dispersing solvent was optimized to 200 μL (FIG. 4).

(6) Optimization of Concentration Conversion of Derivatives

When analyzing polar substances such as glyoxal, methylglyoxal and diacetyl by GC, they are adsorbed on the column or decomposed in the injector, thereby deteriorating the reproducibility of the assay. As shown in FIG. 5, this problem can be solved by converting a polar material into a non-polar material through derivative conversion. Therefore, the conversion efficiencies of glyoxal, methylglyoxal, and diacetyl derivatives for OPD of 1, 5, 10, and 20 g L ^-1 were compared to optimize the concentration of the derivative conversion reagent (OPD). As a result, extraction efficiency of glyoxal for each concentration is 5 g L ^- it was found that the pre-concentration factor (PF) and the extraction recovery (% ER) is maintained from the time ^1. Thus, the concentration of OPD was optimized to 5 g L ^-1 (Figure 6).

(7) Reaction time optimization of derivative conversion reagent

In order to optimize the reaction time of the OPD, the conversion efficiency of the derivatives was compared by varying the reaction time (1, 2, 3 hours) with OPD for glyoxal, methylglyoxal and diacetyl. As a result, the extraction efficiency of glyoxal, methylglyoxal and diacetyl at each time did not show a large difference between the preconcentration factor (PF) and the extraction recovery rate (ER%). Therefore, the reaction time of OPD was optimized to 1 hour (Fig. 7).

(8) Optimization of salt addition

The addition of salt is an important parameter for the extraction efficiency and affects the extraction efficiency to the organic solvent by increasing or decreasing the solubility of the analyte in the aqueous solution. The extraction efficiencies of glyoxal, methylglyoxal and diacetyl were measured for NaCl 0, 2, 4, 6, 8, and 10%. As a result, the preconcentration factor (PF) and the extraction recovery rate (ER%) did not show a significant difference with increasing NaCl concentration (FIG. 8). Thus, the salt concentration was optimized to 0%.

In conclusion, chloroform extraction solvent and methanol were selected as extraction solvent and DLLME extraction condition for analysis of α-dicarbonyl compound in red ginseng concentrate and red ginseng beverage. The volumes of the extraction solvent and the dispersion solvent were optimized to 100 μL and 200 μL, respectively. The concentration of OPD used as a derivative conversion reagent was optimized to 5 g L ^-1 , the conversion time of the derivative reaction time was 1 hour, and the salt concentration was 0%.

(9) Alpha- Dicarbonyl  Validation for compound analysis

A method of analyzing glyoxal, methylglyoxal and diacetyl of red ginseng concentrate and red ginseng beverage by GC-MS was established. Table 1 shows the recovery rates obtained by adding and extracting glyoxal, methylglyoxal, and diacetyl from red ginseng concentrate and red ginseng beverage. Recovery rates of red ginseng concentrate by addition of glyoxal, methylglyoxal and diacetyl standard solutions of 16.7, 33.3, and 166.7 ㎍ L ^-1 were 92.4-96.4%, 97.9-99.6% and 94.4-103.9%, respectively. The recovery rates obtained by adding glyoxal, methylglyoxal, and diacetyl standard solutions of 16.7, 33.3, and 166.7 ㎍ L ^-1 to red ginseng beverages were 99.4-106.9%, 100.3-103.6%, and 103.1-110.7%, respectively. Respectively. The R ² value in the range of 3.3-333.3 μg L ^-1 was 0.9990-0.9992, showing good linearity (Table 2). The precision result by RSD (relative standard deviation) was 2.7-8.2%. The detection limit (Limit of detection, LOD), the limit of quantitation (limit of quantification, LOQ) are each 1.30-1.86 ㎍ L ^- turned out to ^1-1 and 4.33-6.20 ㎍ L. 9 is a chromatogram showing the detection of glyoxal, methylglyoxal, and diacetyl standard solutions using GC-MS.

(10) Red ginseng concentrate and red- Dicarbonyl  Compound analysis

In this study, qualitative and quantitative analysis of glyoxal, methylglyoxal, and diacetyl of red ginseng concentrate and red ginseng beverage was performed through optimized and validated DLLME assay. As a result, the contents of glyoxal, methylglyoxal and diacetyl in 10 red ginseng concentrates were detected as 191.4-4273.6, 1336.4-4798.0, and 0.0-829.8 μg L ^{- 1} , respectively (Table 3, Fig. 10). Further, glyoxal, methyl glyoxal, the content of the diacetyl for 12 ginseng beverages 256.4-864.8, 652.0-3613.0, 19.0-221.4 μg L, ^respectively were detected by ¹ (Table 4, Fig. 11).

[Perilla oil or sesame oil products have α- Dicarbonyl  Detection of compound]

1. Experimental material

(40% in H ₂ O, w / v), MGO (40% in H ₂ O, w / v), DA (97%), 1-methylpyrazole (Q, 99%), 2-methylquinoxaline (2-MQ, 97%), 2,3-dimethylquinoxaline (2,3- DMQ, 97%), sodium dihydrogen phosphate and disodium hydrogen phosphate Were purchased from Sigma-Aldrich (St. Louis, MO, USA). O-phenylenediamine (oPD, 99%) was purchased from Acros Organics (Geel, Belgium). HPLC-grade methanol, chloroform and water were purchased from JT Baker (Phillipsburg, NJ, USA).

2. Experimental Method

(1) sample preprocessing

 Add 0.2 g of oil sample (sesame oil or perilla oil) to a 15 mL glass centrifuge conical tube and mix with 5 mL n-hexane, 5 mL sodium phosphate buffer (0.25 M, pH 7.6), 100 μL 1-MP 1) was added. Then, the above solution was shaken by hand for 2 minutes and centrifuged at 2500 rpm for 5 minutes. After discarding most of the supernatant (organic solvent layer), 3 mL of the lower layer (aqueous layer) was transferred to another glass centrifuge conical tube. For the derivatization, 100 μL OPD solution (10 g L-1) was added and reacted at room temperature for 1 hour.

(2) sesame oil manufacturing

Domestic sesame seeds and perilla seeds were heated at 100 ° C, 150 ° C and 210 ° C for 15 and 30 minutes, respectively, in order to confirm the amount of α-dicarbonyl compounds produced by roasting temperature and time of sesame oil and perilla oil. Sesame oil sesame seeds and perilla seed oil were prepared using commercially available sesame oil weaving machine. The α-dicarbonyl compound of the prepared oil was quantified by optimized DLLME analysis.

(3) DLLME  extraction

After conversion of the derivatives, 1 mL of methanol containing 100 μL of chloroform was quickly injected into the sample solution using a 1.5 mL glass syringe. The sample solution was shaken vigorously for a few seconds and confirmed to be cloudy. After centrifugation at 2500 rpm for 3 minutes, the lower layer was transferred to a 0.15 mL glass insert using a 50 μL glass syringe and analyzed by GC-MS. All experiments were repeated 3 times.

(4) GC -MS condition

The GC-MS analysis was performed using a 7890A gas chromatograph / 5975C mass-selective detector (Agilent Technologies, Palo Alto, Calif., USA) Alto, CA, USA). The inlet temperature was maintained at 250 ° C and the flow rate was maintained at 1.0 mL min -1 in splitless mode. Helium was used as the carrier gas. The purge flow was set to 50 mL min -1, and the purge time was set to 0.75 min. The sample was injected with 1 μL. The oven programming was maintained at 50 &lt; 0 &gt; C for 2 minutes, then increased to 220 &lt; 0 &gt; C at a rate of 10 [deg.] C min-1 and then held for 15 minutes. The temperature of the transfer line, ion source, and quadrupole of the mass spectrometer were maintained at 250, 230, and 150 ° C, respectively. Mass spectra were obtained using an electron ionization of -70 eV. The selected ion monitoring (SIM) mode was used for the identification and quantification of the a-dicarbonyl compound and the internal standard solution (1-MP), and three ions were identified for each analyte (m / z 76, 103 and 130 for Q, m / z 76, 117 and 144 for 2-MQ, m / z 76, 117 and 158 for 2,3-DMQ, ).

(5) Dicarbonyl  The compounds (glyoxal, methylglyoxal, Diacetyl ) For analysis DLLME  Optimize extraction conditions

In order to maximize the efficiency of the DLLME extraction method, several parameters affecting extraction efficiency have been optimized. The optimized parameters are the extraction solvent, the type and volume of the dispersive solvent. Since the? -dicarbonyl compound is a highly active substance, it may be adsorbed to the column by GC analysis or may be lost in the injector, resulting in deterioration of analytical efficiency. In order to solve this problem, glyoxal, methylglyoxal, diacetyl (2-MQ) and 2,3-dimethylquinoxaline (2,3-DMQ), respectively, using o-phenylenediamine (OPD) And analyzed. In order to maximize the conversion efficiency of these derivatives, the amount of o-phenylenediamine (OPD) used as a derivatizing agent and the conversion time of the derivative were tested.

3. Experimental Results

(1) Selection of extraction solvent

For the selection of the extraction solvent, extraction efficiencies were compared using three extraction solvents (chloroform (CHCl ₃ ), chlorobenzene (C ₆ H ₅ Cl), tetrachlorethylene (CH ₂ Cl ₄ )). As a result, when chloroform was used, the peak area of 4-MEI was the highest when chlorobenzene and tetrachlorethylene were used (FIG. 12).

(2) Selection of dispersion solvent

For the selection of the dispersion solvent, extraction efficiencies were compared using three dispersion solvents (methanol (CH ₃ OH), acetonitrile (CH ₃ CN), acetone (C ₃ H ₆ O)). As a result, the extraction efficiency of the three substances (glyoxal, methylglyoxal, diacetyl) was the highest when methanol was used. Therefore, methanol was selected as a dispersion solvent (Fig. 13).

(3) Extraction solvent volume optimization

For the optimization of extraction solvent volumes, extraction efficiencies were determined for chloroform of 100, 150, 200, 250, and 300 μL. As a result, the peak area of 4-MEI was the highest for 100 μL of the extraction solvent. Thus, the volume of the extraction solvent chloroform was optimized to 100 μL (FIG. 14)

(4) Optimization of disperse solvent volume

 To optimize the volume of the dispersion solvent, the extraction efficiency was measured for 0.2, 0.4, 0.6, 0.8, and 1.0 mL of methanol. As a result, the peak area value was highest at 1.0 mL. Thus, the volume of the dispersed solvent was optimized to 1.0 mL. (Fig. 15)

(5) Optimization of derivative conversion reagent mass

In order to optimize the mass of the OPD, the conversion efficiencies of 200, 500, 1000, 2000, 5000 and 10,000 μg of OPD were compared. As a result, the highest conversion efficiency was obtained when 1000 μg of OPD was used. Thus, the mass of OPD was optimized to 1000 μg (FIG. 16).

(6) Optimization of derivative conversion time

In order to optimize the conversion time of the derivatives, the conversion efficiencies of the derivatives for 30 minutes, 1, 2, and 3 hours were compared. As a result, there was no significant difference in the conversion efficiency of methylglyoxal and diacetyl with time (P <0.05), and in case of glyoxal, the conversion efficiency of the derivatives was significantly different between 30 minutes and 1 hour (P <0.001), but there was no significant difference after one hour. Thus, the derivative conversion time was optimized to one hour. (Fig. 17).

(7) Dicarbonyl  Validation for compound analysis

A method for analyzing? -Dicarbonyl compounds in sesame oil was established. Table 5 shows the recovery rates obtained by adding &amp;alpha; -dicarbonyl compound to sesame oil. The recovery rates obtained by adding glyoxal, methylglyoxal, diacetyl standard solutions of 250 and 1000 ng g ^-1 to sesame oil were 100.6 ~ 115.4% and 106.4 ~ 112.0%, respectively (Table 5).

In addition, the R2 value ranged from 0.9988 to 0.9995 with good linearity in the range of 0.2-100 ng mL ^-1 . The accuracy of the relative standard deviation (RSD) was 0.5-9.6% for Intra-day and 1.3-9.8% for Inter-day. The detection limit (Limit of detection, LOD), the limit of quantitation (limit of quantification, LOQ) are respectively 0.22 ~ 0.86 ng mL ^{- 1} and was 0.66 ~ 2.60 ng mL ^-1 (Table 6).

18 is a chromatogram showing the detection of an? -Dicarbonyl compound standard solution using GC-MS.

(8) α- Dicarbonyl  Compound quantitative analysis

Through optimization and validated assays, alpha-dicarbonyl compounds of sesame oil and perilla oil were quantitatively analyzed. As a result, in sesame oil, glyoxal, methylglyoxal and diacetyl were quantified respectively as 0 to 175.4 ng g ^-1 , 0 to 990.5 ng g ^-1 and 0 to 220.9 ng g ^-1 , and in the perilla oil, glyoxal, methylglyoxal , And diacetyl were quantified in the range of 0 to 96.4 ng g ^-1 , 0 to 410.8 ng g ^-1 , and 0 to 197.5 ng g ^-1, respectively. In particular, no α-dicarbonyl compound was detected in raw sesame oil (sample No. 8) and raw oil (sample No. 3) (Table 7).

FIG. 19 shows a chromatogram of sesame oil sample 1 analyzed by DLLME.

(9) Roasting temperature of sesame oil and perilla oil. Dicarbonyl  Compound yield analysis

In order to confirm the amount of α-dicarbonyl compound produced by roasting temperature and time of sesame oil and perilla oil, the sesame and perilla seeds were heated at 100, 150 and 210 ℃ for 15 and 30 minutes, respectively, and then α-dicarbonyl compound was optimized And analyzed by the DLLME method. As a result, when sesame and perilla seeds were roasted at 210 DEG C for 30 minutes, the amount of the [alpha] -dicarbonyl compound tended to increase sharply as compared to unroasted sesame oil and perilla oil (Fig. 20).

Claims (6)

A method for determining whether an alpha-dicarbonyl compound is contained in a sample, comprising the steps of:
(S1) adding o-phenylenediamine dihydrochloride (OPD) to a sample and reacting;
(S2) adding an extraction solvent and a dispersion solvent according to a dispersive liquid-liquid microextraction (DLLME) method and centrifuging; And
(S3) analyzing the alpha-dicarbonyl compound in the extraction solution with a gas chromatograph-mass spectrometer,
Chloroform in the extraction solvent, and methanol in the dispersion solvent. The method of claim 1, wherein the sample is a red ginseng product, a perilla oil product, or a sesame oil product. The method according to claim 1, further comprising the step of adding an internal standard material to the sample in step (S1), wherein the internal standard material is 1-methylpyrazole (1-MP) Lt; / RTI &gt; The process according to claim 1, wherein the alpha-dicarbonyl compound is glyoxal, methylglyoxal or diacetyl. The method of claim 1, wherein the ion (ion, m / z) of quinoxaline in step (S3) 130, the ion (m / z) of methylquinoxaline is 144, Or the ion (ion, m / z) of 2,3-dimethylquinoxaline is 158. The process according to claim 1, wherein the volume of the dispersing solvent to the volume of the extracting solvent is 1: 1-15.

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