KR20230096309A - Method of analyzing aroma constituent of pericarp - Google Patents

Abstract

The present invention relates to a method for analyzing the aroma components of fruit peel, which comprises: (A)a step of freeze-dried fruit peel; (B) a step of pulverizing the freeze-dried fruit peel and adsorbing and extracting the freeze-dried fruit peel through an adsorbent in a solid-phase microextraction arrow (SPME-Arrow) method; and (C) a step of placing the adsorbent in a gas chromatography-mass spectrometry (GC-MS) and desorbing and analyzing volatile aroma components adsorbed and extracted by the adsorbent. Accordingly, the aroma components of fruit peel can be analyzed with high accuracy.

Description

Method of analyzing aroma constituent of pericarp of citrus peel for development of natural food flavor {Method of analyzing aroma constituent of pericarp}

The present invention relates to a method capable of analyzing fragrance components of fruit peel accurately at a high level and analyzing various fragrance components.

Gas chromatography (GC), one of the conventional representative analysis methods, has the disadvantage of using dangerous or highly toxic organic solvents in order to extract and concentrate components to be analyzed from a sample. In particular, when a polar compound present in a small amount in an aqueous solution is concentrated by vaporizing an organic solvent, sample loss occurs.

In order to overcome this, examples using solid-phase microextraction (SPME), which can increase sensitivity by improving the detection limit of trace components without using a solvent, have been reported.

For example, a method in which an analyte is extracted by solid-phase microextraction (SPME) and then analyzed by gas chromatography-mass spectrometry (GC-MS). The SPME utilizes the adsorption of vaporized components of the sample to the fiber when a fiber having an adsorbent fixed thereto is coupled to a thin needle, and then the needle is passed through a sample vial to expose the fiber to the gas phase.

However, the extraction by SPME is suitable for volatile substances, but in the case of weakly volatile compounds, it is difficult to evaporate into the headspace and has a disadvantage of low analytical sensitivity.

In addition, it is difficult to accurately analyze the fragrance component in the case of extraction by SPME due to the nature of the fragrance component having a low content is difficult to concentrate.

On the other hand, flavoring plays a very important role in food, cosmetics, beverages, surfactants, feeds, chemicals, biopharmaceuticals, and pharmaceutical industries. The production method of such a fragrance is largely divided into a chemical synthesis method and a biological method. Production of a fragrance ingredient through a chemical synthesis method is capable of mass production of fragrance with various types of chemical structures, but it is possible to produce an undesirable racemic mixture. Not only is it possible to make it, but it also has the disadvantage of low substrate specificity.

In addition, consumers' consumption propensity is changing in the direction of preferring foods, medicines, chemical products, and pharmaceutical products that are environmentally friendly and contain fragrance ingredients that are harmless to the human body. There is a growing interest in fragrance ingredients produced by biological methods.

Currently, the size of the spice market is about 10 billion dollars worldwide, and the domestic market is about 130 million dollars. It is somewhat dependent on income.

In particular, despite the fact that domestically consumed fragrances (citrus-type) are mainly imported, existing studies conducted in Korea have focused on herbs derived from domestic agricultural products, legumes, medicinal and herbal medicinal plants, and natural fragrances using domestic materials such as citrus fruits is not being developed.

Therefore, there is a need for a method capable of accurately analyzing fragrance components at a high level in order to develop excellent natural fragrances.

Republic of Korea Patent No. 2055499 Republic of Korea Patent No. 2124999

An object of the present invention is to provide a method that can accurately analyze the fragrance components of the peel at a high level and analyze various fragrance components.

In addition, another object of the present invention is to provide a perfume composition comprising the fragrance component of the peel analyzed according to the above method.

In addition, another object of the present invention is to provide a food composition containing the fragrance component of the peel analyzed according to the above method.

In addition, another object of the present invention is to provide a cosmetic composition containing the fragrance component of the peel analyzed according to the above method.

Method for analyzing the fragrance component of the peel of the present invention for achieving the above object is (A) freeze-drying the peel; (B) adsorbing and extracting the lyophilized rind through an adsorbent using a solid-phase microextraction arrow (SPME-Arrow) after crushing the lyophilized rind; and (C) putting the adsorbent into gas chromatography-mass spectrometry (GC-MS) and desorbing and analyzing volatile fragrance components adsorbed and extracted from the adsorbent.

Freeze-drying in step (A) may be performed for 15 to 25 hours by freezing at -80 to -70 ° C.

In step (B), the adsorbent may be divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS).

In step (B), the extraction may be performed at an extraction temperature of 35 to 55 °C, an extraction time of 50 to 70 minutes, and an equilibrium temperature of 5 to 25 minutes.

The area value of the analyzed pericarp fragrance component may be 1.0X10 ¹¹ to 9.0X10 ¹¹ .

The main aroma components of the peel analyzed above are β-Myrcene, D-Limonene, γ-Terpinene, p-cymene, and Terpi Terpinolen, Copaene, β-elemene, α-Humulene, α-Farnesene and Linalool,

Each area value of the main fragrance component is 1.0X10 ¹⁰ to 6.0X10 ¹⁰ of β-Myrcene, 1.0X10 ¹¹ to 5.0X10 ¹¹ of D-Limonene, 5.0X10 ¹⁰ to 9.9X10 ¹⁰ of γ-terpinene, 1.0X10 ¹⁰ to 5.0 X10 ¹⁰ of p-cymene (p-cymene), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Terpinolen (Terpinolen), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Copaene (Copaene), 1.0X10 ¹⁰ to 7.0X10 ¹⁰ of β-elemene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-Humulene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-farnesene and 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of linalool.

The total area value of the main fragrance component of the analyzed pericarp may be 1.0X10 ¹¹ to 7.0X10 ¹¹ .

The rind may be a citrus rind.

The citrus fruit may be citrus (Citrus unshiu), lemon (Citrus limon), orange (Citrus sinensis), citron (Citrus junos), grapefruit (Citrus paradisi), or hallabong (Citrus hybrid Shiranuhi).

The analyzed fragrance component may have an accuracy of 80% or more during validation.

In addition, the fragrance composition of the present invention for achieving the above other object may include the fragrance component of the peel analyzed by the above analysis method.

In addition, the food composition of the present invention for achieving the above another object may include the fragrance component of the peel analyzed by the above analysis method.

In addition, the cosmetic composition of the present invention for achieving the above another object may include the fragrance component of the peel analyzed by the above analysis method.

The fragrance component of the peel analyzed according to the analysis method of the present invention can be analyzed with high accuracy as well as various fragrance components.

Figure 1a is a GC graph analyzing fragrance components according to Example 1, Figure 1b is a GC graph analyzing fragrance components according to Comparative Example 16, Figure 1c is a GC graph analyzing fragrance components according to Comparative Example 17 , FIG. 1D is a GC graph analyzing fragrance components according to Comparative Example 18, and FIG. 1E is a GC graph analyzing fragrance components according to Comparative Example 19.
2 is a GC graph in which fragrance components are analyzed according to Comparative Example 20.

The present invention relates to a method capable of analyzing fragrance components of fruit peel accurately at a high level and analyzing various fragrance components.

Hereinafter, the present invention will be described in detail.

The method for analyzing the fragrance component of the peel of the present invention comprises the steps of (A) freeze-drying the peel; (B) adsorbing and extracting the lyophilized rind through an adsorbent using a solid-phase microextraction arrow (SPME-Arrow) after crushing the lyophilized rind; and (C) putting the adsorbent into gas chromatography-mass spectrometry (GC-MS) and desorbing and analyzing volatile fragrance components adsorbed and extracted from the adsorbent.

First, in step (A), the skin is frozen at -80 to -70 ° C and freeze-dried for 15 to 25 hours, preferably 20 to 24 hours.

Freeze-drying is to extract a large amount of fragrance components during extraction, and when freeze-drying is not performed or other drying methods such as hot air drying, natural drying, cold air drying, low-temperature vacuum drying, and superheated steam drying are performed, the desired Fragrance components cannot be obtained, and fragrance components are not extracted in large quantities.

If the freeze-drying time is less than the lower limit, the desired fragrance component cannot be obtained, and if it exceeds the upper limit, the fragrance component is not extracted in large quantities.

The peel used in the present invention is a citrus peel, and the citrus peel is citrus (Citrus unshiu), lemon (Citrus limon), orange (Citrus sinensis), citron (Citrus junos), grapefruit (Citrus paradisi) or hallabong (Citrus hybrid) Shiranuhi).

Next, in step (B), the lyophilized rind is pulverized, and then the lyophilized rind is adsorbed and extracted through an adsorbent using a solid-phase microextraction arrow (SPME-Arrow), In the step (C), the adsorbent is put into a gas chromatography-mass spectrometer (GC-MS), and volatile fragrance components adsorbed and extracted by the adsorbent are desorbed and analyzed.

The analysis method of the present invention may be performed using equipment in which a solid-phase microextraction arrow (SPME-Arrow) device is coupled to a gas chromatography-mass spectrometer (GC-MS). Specifically, after extracting an analyte by adsorption from the lyophilized skin using a SPME-Arrow instrument, the extracted analyte is analyzed using a gas chromatography-mass spectrometer (GC-MS).

In the present invention, when SPME is used rather than solid-phase microextraction Arrow (SPME-Arrow), the number of fragrance components detected and the peaks detected by gas chromatography-mass spectrometry (GC-MS) The area value is small, and the accuracy is less than 50% even in validation.

A fiber coated with an adsorbent is introduced into a vial containing the freeze-dried pericarp, and the fragrance component of the pericarp is adsorbed and extracted by the adsorbent. The adsorbent is not particularly limited, but divinylbenzene / carboxen / polydimethylsiloxane (Divinylbenzene / Carboxen / Polydimethylsiloxane , DVB/CAR/PDMS) can be used.

As the adsorbent, polydimethylsiloxane (PDMS), polydimethylsiloxane / divinylbenzene (DVB / PDMS), carboxen / polydimethylsiloxane (CAR / PDMS), polydimethylsiloxane ( PDMS) or polyacrylate (PA) is not preferable because the number of fragrance components and peak area values may be low.

Extraction with the adsorbent is performed at an extraction temperature of 35 to 55 °C, preferably 40 to 45 °C; extraction time of 50 to 70 minutes, preferably 55 to 60 minutes; and an equilibrium temperature of 5 to 25 minutes, preferably 10 to 15 minutes; It is preferable to detect a large number of fragrance components and a high peak area value.

When the extraction temperature, extraction time, and parallel temperature are out of the above ranges, the number of fragrance components to be analyzed is rapidly reduced, and the peak area value may be significantly lowered.

The area value of the peak detected by gas chromatography-mass spectrometry (GC-MS) for all flavor components of the peel analyzed according to the present invention is 1.0X10 ¹¹ to 9.0X10 ¹¹ .

In particular, each area value of the main fragrance component of the analyzed pericarp is 1.0X10 ¹⁰ to 6.0X10 ¹⁰ β-Myrcene, 1.0X10 ¹¹ to 5.0X10 ¹¹ of D-Limonene, 5.0X10 ¹⁰ to 9.9X10 ¹⁰ of γ-terpinene, 1.0X10 ¹⁰ to 5.0 X10 ¹⁰ of p-cymene (p-cymene), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Terpinolen (Terpinolen), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Copaene (Copaene), 1.0X10 ¹⁰ to 7.0X10 ¹⁰ of β-elemene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-Humulene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-farnesene and 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Linalool.

The total peak area values of the main fragrance components of the peel are 1.0X10 ¹¹ to 7.0X10 ¹¹ , showing high peak area values.

The fragrance component of the peel analyzed by the analysis method of the present invention is accurately detected at a high level of 80% or more during validation.

In addition, the present invention can provide a food composition containing the analyzed fragrance component.

The food composition is not particularly limited as long as it is a product that can use a fragrance component, but preferably includes soft drinks, fermented milk, bread, noodles, confectionery, frozen confectionery, sausages, cheese, edible processed foods, or processed marine products.

In addition, the present invention can provide a health functional food containing the analyzed fragrance component.

The 'food composition' includes, in addition to fragrance components, food raw materials that can be used as foods described in the food standards and specifications ('Food Code') commonly used in food manufacturing, and food additives described in the Food Additives Code.

It does not need to be particularly limited, but includes, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. The carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sugar, lactose and the like; oligosaccharides or polysaccharides such as dextrin, starch syrup, cyclodextrin and the like; Sugar alcohols such as xylitol, sorbitol, erythritol and the like can be used. As the flavoring agent, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be used.

When preparing a food composition containing the fragrance component, the fragrance component does not need to be particularly limited as long as a large amount of the fragrance component can last for a long time, but, for example, 0.1 to 99% by weight, 0.5 to 95% by weight, 1 to 90% by weight, 1 to 80% by weight, 1 to 70% by weight, 1 to 60% by weight, or 1 to 50% by weight.

In the food composition, the fragrance component varies depending on the condition, body weight, presence or absence or degree and period of disease of the eater, but may be appropriately selected by a person skilled in the art. For example, it may be 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, still more preferably 20 to 800 mg, and most preferably 50 to 500 mg based on the daily dose. There is, and the number of administrations need not be particularly limited, but can be adjusted by a person skilled in the art within the range of three times a day to once a week.

The food composition does not need to be particularly limited, but may be, for example, a powder, granule, tablet, capsule, pill, extract, jelly formulation, tea bag formulation or beverage formulation.

In addition, fragrance components can be given to general foods, and foods that can be added do not need to be particularly limited, but for example, confectionery and bread exemplified in the food standards and specifications ('Food Code') according to Article 7 of the Food Sanitation Act or rice cakes, processed cocoa products or chocolates, processed meat or egg products, processed fish meat products, tofu or jelly, noodles, teas, coffee, beverages, special purpose foods, soybeans, seasonings, dressings, kimchi, salted seafood, pickled foods, It can be added to stewed food, alcoholic beverages, raisins, and other foods. In addition, it can be added to dairy products, processed meat products, packaged meat, and egg products exemplified in the processing standards and ingredient specifications of livestock products ('livestock product code') according to Article 4 of the Livestock Products Sanitation Control Act.

The above ‘functional health food’ refers to food manufactured (including processing) in accordance with legal standards using raw materials or ingredients that have functional properties useful for the human body (Article 3, Subparagraph 1 of the Health Functional Food Act). The term 'functional health food' may differ from country to country, but 'Dietary Supplement' in the US, 'Food Supplement' in Europe, 'Health Functional Food' or 'Health Functional Food' in Japan. It may correspond to 'Food for Special Health Use (FoSHU)' or 'Health Food' in China.

The food composition or health functional food may additionally contain food additives, and the suitability as a food additive is determined according to the standards and standards for the item in accordance with the general rules and general test methods of the 'Food Additive Code' unless otherwise specified. follow

Hereinafter, preferred embodiments are presented to aid understanding of the present invention, but the following examples are merely illustrative of the present invention, and various changes and modifications are possible within the scope and spirit of the present invention. It is obvious to those skilled in the art, It goes without saying that these variations and modifications fall within the scope of the appended claims.

[By skin drying method]

Example 1.

The ultrasonically washed tangerine peel was freeze-dried at -78 ° C for 24 hours, and then the freeze-dried peel was pulverized and divinylbenzene / divinylbenzene / The freeze-dried skin was adsorbed and extracted through a carboxen/polydimethylsiloxane (Divinylbenzene/Carboxen/Polydimethylsiloxane, DVB/CAR/PDMS) adsorbent. Thereafter, the adsorbent was put into a gas chromatography-mass spectrometry (GC-MS), and the volatile fragrance component adsorbed and extracted by the adsorbent was desorbed to analyze the fragrance component.

The extraction was performed at an extraction temperature of 45 °C, an extraction time of 60 minutes, and an equilibrium temperature of 15 minutes.

Specifically, 0.1 g of the lyophilized peel was placed in 20 mL headspace vials (Wheaton, Millville, USA), sealed with a dedicated screw cap, and Ethyl acetate-d ₈ (10 μg/mL, 10 μL) as an internal standard before extraction. , 1-hexyl alcohol-d ₁₃ (1 μg/mL, 10 μL), 3-octanone (1 μg/mL, 10 μL), trimethylacetaldehyde (10 μg/mL, 10 μL), 3,4-dimethylphenol (1 μg/mL) mL, 10 μL) and toluene-d ₈ (1 μg/mL, 10 μL) solutions were added to the vial containing the sample. For instrumental analysis, a Trace 1310 gas chromatograph equipped with SPME-Arrow and a TSQ 9000 TQMS (Thermo Scientific, Waltham, USA) were used, and the samples were stored on a DB-WAX column (60 m X 0.25 mm id, 0.25 μm film thickness, Agilent Technologies). The carrier gas was flowed at He 1.0 mL/min in constant flow mode, the temperature of the GC inlet was 220 °C, and the split mode (split ratio; 5: 1) was set. GC oven programming was maintained at 45 ℃ for 10 minutes, then raised to 100 ℃ at 2 ℃ per minute, and then raised to 230 ℃ at 3 ℃ per minute. The MS transfer line and MSD source temperatures were maintained at 230 and 280 °C, respectively, the detector mode was set to scan mode, and the scan range was set to 30-550 m/z .

Comparative Example 1. Hot Air Drying

It was carried out in the same manner as in Example 1, but instead of freeze-drying, the skin was dried in hot air at 70 ° C. for 6 hours, and the fragrance component was analyzed using this.

Comparative Example 2. Superheated Steam Drying

It was carried out in the same manner as in Example 1, but instead of freeze-drying, the skin was dried for 20 minutes under an oven temperature of 100 ° C and a steam temperature of 250 ° C, and then the fragrance component was analyzed using this.

Comparative Example 3. Low-temperature vacuum drying

It was carried out in the same manner as in Example 1, but instead of freeze-drying, it was dried in a low-temperature vacuum state at 50 ° C. for 24 hours, and then the fragrance component was analyzed using this.

<Test Example Ⅰ>

Test Example 1. Fragrance component analysis

The results of analyzing the fragrance components according to Example 1 and Comparative Examples 1 to 3 are shown in Table 1 below.

division µg/mL Odor description Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Alcohols Ethanol 40,772,468±750,261 - - - alcohol, floral, sweet 2-Nonen-1-ol 137,842,186±2,508,591 244,295,652±7,941,577 176,763,409±6,350,746 215,253,295±1,613,550 green 3-Hexen-1-ol 138,624,538±6,616,980 - - - grass Linalool 28,434,761,206±1,034,064,339 18,197,911,208±908,070,104 16,470,459,722±259,513,051 9,536,564,026±125,755,163 bergamot, grape, lavender, rose 4-Thujanol 477,540,031±6,042,621 76,547,673±1,702,745 - - - α-Terpineol 7,572,309,076±48,554,534 2,390,008,569±179,725,904 2,015,533,203±22,437,175 1,499,773,442±52,148,599 lily, mint, oil, sweet trans-Carveol 465,882,686±15,002,135 137,853,340±2,538,334 170,710,042±791,852 161,820,136±5,469,867 caraway, spearmint Carveol 139,428,987±7,602,405 62,763,493±1,799,695 71,981,822±2,879,057 58,580,533±904,933 caraway, fresh, spearmint Benzyl alcohol - 53,722,426±2,563,919 - 68,181,502±1,189,701 almond,floral,roasted bread Perillic alcohol 1,097,876,074±8,643,741 150,817,682±10,605,179 118,543,097±3,403,516 169,334,353±4,298,943 green, pungent, sweet Elemol 1,396,690,778±8,647,753 204,026,071±15,355,432 324,968,216±11,889,734 234,911,445±5,855,511 green wood Aldehydes Octanal 252,167,262±6,824,339 - - - citrus, green, nut, pungent Hexyl formate 49,971,824±1,367,388 - - - - Nonanal 2,446,036,157±33,775,389 2,262,509,641±87,675,617 3,152,935,915±124,634,268 1,622,096,548±27,175,850 citrus, floral, green Citronellal 2,180,371,045±49,898,314 1,698,228,181±42,690,524 2,492,407,700±47,230,228 1,150,766,242±51,663,810 Citrus, fat, leaf Decanal - 9,639,804,616±156,457,208 - - floral, fried, orange peel Dodecanal 1,097,876,074±72,450,502 523,292,483±5,132,989 877,209,149±11,348,326 - Citrus, fat, lily Perill aldehyde 1,396,690,778±61,094,883 418,960,450±5,202,435 528,593,122±5,720,632 239,799,116±10,844,942 green, pungent, sour, spice 2,4-Decadienal - 88,062,485±1,891,170 - - deep fried, seaweed, sweet Ester Nonyl acetate 378,598,593±2,121,657 110,234,193±649,846 - - fruit, sweet Citronellol acetate 1,061,449,497±73,350,913 512,537,524±11,248,110 714,084,133±11,698,375 625,771,776±8,360,931 floral, fragrant, rose Geranyl acetate 2,576,585,674±118,965,319 1,102,967,242±21,170,812 1,615,738,433±38,228,120 1,280,704,610±54,509,313 lavender, rose, sweet Limonen-10-ylacetate 975,221,287±33,969,434 455,951,684±8,477,661 643,606,509±5,979,118 410,423,363±16,465,255 fruit, sweet Perillyl acetate 321,363,786±14,542,259 148,144,797±18,977,291 242,856,759±8,622,103 152,971,362±1,357,916 - Hydrocarbons 2-Pinene 5,889,654,352±156,809,503 139,295,172,693±5,630,076,630 65,876,706,759±1,247,998,029 119,495,317,055±9,469,137,793 cedarwood, pine, resin, sharp Sabinen - 5,609,839,653±588,510,656 46,595,780,918±3,708,178,091 - herb, pepper, turpentine, wood β-Myrcene 15,357,593,738±517,844,786 44,410,200,415±2,310,094,747 26,686,565,362±1,730,909,553 41,783,656,675±1,611,217,250 balsamic, fruit, herb, must D-Limonene 144,434,403,863±3,711,141,252 169,121,553,931±3,456,057,674 104,427,536,406±3,120,224,496 94,291,820,391±1,345,168,221 citrus, mint β-Thujene 2,393,772,838 ±57,411,780 - - - γ-Terpinene 83,909,651,438±2,011,974,394 44,242,082,933±334,069,423 60,945,774,213±1,545,256,612 55,243,258,919±3,033,987,800 bitter, citrus, resin, turpentine p-Cymene 17,716,967,701±243,527,491 19,666,239,938±151,616,574 20,985,694,524±295,332,055 20,369,218,187±644,917,142 citrus, fresh, solvent, wood Terpinolen 17,714,852,752±152,485,814 25,657,795,042±268,579,248 23,621,330,598±282,991,893 24,947,737,339±1,159,856,285 pine, plastic, sweet Tridecane 185,381,524±6,893,531 - - - alkane Allocimene 309,860,608±6,821,151 - 374,814,356±8,474,117 363,309,364±8,525,838 α-Cubebene 4,233,386,214±68,867,347 - - - herb, wax, wood δ-Elemene 12,601,924,044±545,639,978 6,326,118,492±44,321,729 9,483,575,719±227,295,202 6,091,733,063±117,286,427 sweet, wood Copaene 13,256,889,120±298,349,198 - 11,348,382,896±288,842,283 9,017,160,318±238,062,483 spice, wood Ylangene - 45,821,446±4,672,306 - - fruit β-Elemene 32,428,844,914±1,194,049,841 18,189,468,707±278,279,855 21,667,502,671±205,391,090 18,432,134,935±416,626,763 fresh, fruit, herb, sweet, wax Caryophyllene 8,934,502,676±339,215,294 3,608,667,174±74,673,851 5,196,050,993±96,772,328 4,306,534,392±92,217,360 fried, spice, wood Germacrene B 2,717,131,008±132,510,621 971,081,744±3,852,583 1,499,425,576±20,661,150 - balsamic, mushroom, spice, wood α-Humulene 11,231,029,489±717,919,864 5,083,603,563±80,597,622 6,353,475,520±123,568,422 5,792,578,632±96,467,783 balsamic, hop, spice, wood γ-Muurolene 1,780,459,990±66,660,160 509,031,782±20,274,424 1,008,700,674±4,590,390 752,500,639±22,196,917 herb, spice, wood (-)-Germacrene D 8,284,683,255±36,290,889 4,346,302,454±50,201,849 5,160,746,827±133,281,036 4,606,924,291±142,834,927 spice, wood α-Selinene 3,538,958,595±157,585,393 1,094,827,375±6,759,655 1,674,639,771±36,282,239 1,556,647,528±33,630,136 wood α-Farnesene 21,166,542,710±1,044,561,483 12,577,252,937±830,506,956 12,862,766,742±194,283,681 11,167,424,836±130,895,606 citrus, floral, sweet, wood δ-Cadinene 7,425,906,862±208,270,739 2,744,530,810±11,007,104 4,165,988,331±37,253,051 3,211,044,617±190,260,816 medicine, thyme, wood γ-Cadinene 1,587,851,430±41,255,526 411,228,737±6,431,855 - 641,260,183±10,737,222 wood Cubenene - 355,335,718±4,017,315 731,643,714±10,144,777 585,579,884±14,964,486 fruit, spice α-Cadinene 942,110,391±31,236,741 300,577,405±22,182,240 528,832,519±5,528,279 277,824,447±207,366,495 medicine, thyme, wood trans-γ-Bisabolene 481,054,090±26,880,979 136,916,909±2,985,082 254,499,441±6,129,672 174,438,378±9,786,245 ketones Sulcatone 60,044,941±657,375 - - - citrus, mushroom, strawberry Oxides (+)-Limonene oxide 601,367,815±15,320,846 1,367,718,449±37,158,195 877,915,358±25,603,207 - fruit, green, spice total peak area value 470,117,445,946 444,650,007,716 461,944,741,119 440,535,055,820 - number of peaks 47 43 38 36 -

As shown in Table 1 above, it was confirmed that the fragrance component analyzed according to Example 1 of the present invention had a higher content (total peak area value was higher) than Comparative Examples 1 to 3, compared to Comparative Examples 1 to 3. It was confirmed that various fragrance components were analyzed.

Test Example 2. Analysis of major fragrance components

Among the fragrance components analyzed in Test Example 1, only the main fragrance components were summarized.

division µg/mL Odor description Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 β-Myrcene 15,357,593,738±517,844,786 44,410,200,415±2,310,094,747 26,686,565,362±1,730,909,553 41,783,656,675±1,611,217,250 balsamic, fruit, herb, must D-Limonene 144,434,403,863±3,711,141,252 169,121,553,931±3,456,057,674 104,427,536,406±3,120,224,496 94,291,820,391±1,345,168,221 citrus, mint γ-Terpinene 83,909,651,438±2,011,974,394 44,242,082,933±334,069,423 60,945,774,213±1,545,256,612 55,243,258,919±3,033,987,800 bitter, citrus, resin, turpentine p-cymene (p-cymene) 17,716,967,701±243,527,491 19,666,239,938±151,616,574 20,985,694,524±295,332,055 20,369,218,187±644,917,142 citrus, fresh, solvent, wood Terpinolen 17,714,852,752±152,485,814 25,657,795,042±268,579,248 23,621,330,598±282,991,893 24,947,737,339±1,159,856,285 pine, plastic, sweet Copaene 13,256,889,120±298,349,198 - 11,348,382,896±288,842,283 9,017,160,318±238,062,483 spice, wood β-elemene 32,428,844,914±1,194,049,841 18,189,468,707±278,279,855 21,667,502,671±205,391,090 18,432,134,935±416,626,763 fresh, fruit, herb, sweet, wax α-Humulene 11,231,029,489±717,919,864 5,083,603,563±80,597,622 6,353,475,520±123,568,422 5,792,578,632±96,467,783 balsamic, hop, spice, wood α-Farnesine (α-Farnesene) 21,166,542,710±1,044,561,483 12,577,252,937±830,506,956 12,862,766,742±194,283,681 11,167,424,836±130,895,606 citrus, floral, sweet, wood Linalool 28,434,761,206±1,034,064,339 18,197,911,208±908,070,104 16,470,459,722±259,513,051 9,536,564,026±125,755,163 bergamot, grape, lavender, rose total peak area value 385,651,536,931 357,146,108,674 305,369,488,654 290,581,554,258 -

As shown in Table 2 above, the main fragrance components analyzed according to Example 1 of the present invention had a higher total peak area value than Comparative Examples 1 to 3, so it was confirmed that the content of the main fragrance components was high. If a large amount of this is analyzed, the accuracy of the fragrance component is improved accordingly, so the peak area value is an important indicator.

Test Example 3. Fragrance component verification

It was conducted to verify the assay method according to Example 1 of the present invention. Linearity, detection limit, quantification limit, intraday precision, daily precision, and accuracy for the main fragrance components analyzed in Test Example 2 were confirmed.

For linearity, a calibration curve was prepared in the concentration range of 100 to 5000 ng/mL, and a correlation coefficient of 0.99 or higher was confirmed; The limits of detection and quantification were set at the S/N ratio of 3.3 or more and the quantification limit of 10 or more.

In addition, intraday precision was analyzed 5 times a day to confirm a value of less than 20% of the relative standard deviation; The daily precision was analyzed once a day for 5 days and a value of 20% or less of the relative standard deviation was confirmed; Accuracy was confirmed as a value of 80 to 120% in the recovery rate of the target concentration.

division coefficient of determination
(r ² ) detection limit
(ng/mL) Limit of quantification (ng/mL) intraday precision
(%) daily precision
(%) accuracy
(%) β-Myrcene 0.9997 0.26 0.79 6.72 3.31 94.96 D-Limonene 0.9998 0.39 1.20 4.41 6.76 97.40 γ-Terpinene 0.9999 0.46 1.39 4.17 3.44 99.75 p-cymene (p-cymene) 0.9997 0.12 0.38 4.36 1.32 98.23 Terpinolen 0.9996 0.77 2.33 3.41 4.66 95.66 Copaene 0.9986 0.10 0.31 4.51 3.09 95.60 β-elemene 0.9996 0.76 2.29 7.89 5.95 94.11 α-Humulene 0.9994 0.28 0.86 2.05 5.72 98.50 α-Farnesine (α-Farnesene) 0.9991 0.10 0.32 8.05 6.51 94.90 Linalool 0.9996 0.19 0.57 9.57 11.20 99.05

As shown in Table 3 above, it was confirmed that the verification of the main fragrance components analyzed according to Example 1 of the present invention showed a high accuracy of 94.0 to 99.5%.

[By extraction conditions]

Example 1.

In Example 1, extraction was performed at an extraction temperature of 45 °C, an extraction time of 60 minutes, and an equilibrium temperature of 15 minutes.

Comparative Examples 4 to 15.

It was carried out in the same manner as in Example 1, but the fragrance components were analyzed by extraction according to the conditions of Table 4 below.

<Test Example Ⅱ>

Test Example 4. Fragrance component analysis

The results of analyzing the fragrance components according to Example 1 and Comparative Examples 4 to 15 are shown in Table 4 below.

division Extraction temperature (℃) Extraction time (minutes) Equilibration time (minutes) total peak area value Example 1 45 60 15 470,117,445,946 Comparative Example 4 45 30 5 141,769,520,951 Comparative Example 5 35 30 15 154,004,565,596 Comparative Example 6 35 60 25 155,414,152,995 Comparative Example 7 45 30 25 257,976,061,711 Comparative Example 8 35 60 5 159,102,793,187 Comparative Example 9 55 30 15 162,218,073,172 Comparative Example 10 35 90 15 162,762,978,739 Comparative Example 11 45 90 5 64,779,206,613 Comparative Example 12 55 60 5 65,285,482,917 Comparative Example 13 55 60 25 268,475,976,369 Comparative Example 14 55 90 15 169,224,234,834 Comparative Example 15 45 90 25 274,010,834,104

As shown in Table 4 above, the fragrance components analyzed according to Example 1 of the present invention had a higher total peak area value than Comparative Examples 4 to 15, so it was confirmed that the content of fragrance components was high.

[By adsorbent type and extraction device]

Example 1.

The above Example 1 was analyzed using divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS) as an adsorbent.

Comparative Example 16. Use of polydimethylsiloxane/divinylbenzene (DVB/PDMS)

It was carried out in the same manner as in Example 1, but the fragrance component was analyzed using polydimethylsiloxane/divinylbenzene (DVB/PDMS) as an adsorbent.

Comparative Example 17. Carboxen/polydimethylsiloxane (CAR/PDMS)

Carboxen/polydimethylsiloxane (CAR/PDMS) was used as an adsorbent, but the fragrance component was analyzed in the same manner as in Example 1.

Comparative Example 18. Polydimethylsiloxane (PDMS)

It was carried out in the same manner as in Example 1, but the fragrance component was analyzed using polydimethylsiloxane (PDMS) as an adsorbent.

Comparative Example 19. Polyacrylate (PA)

It was carried out in the same manner as in Example 1, but the fragrance component was analyzed using polyacrylate (PA) as an adsorbent.

Comparative Example 20. Use of SPME

It was carried out in the same manner as in Example 1, but the fragrance components were analyzed using solid-phase microextraction (SPME) instead of SPME-Arrow.

<Test Example Ⅲ>

Test Example 5. Fragrance component analysis

Figure 1a is a GC graph analyzing fragrance components according to Example 1, Figure 1b is a GC graph analyzing fragrance components according to Comparative Example 16, Figure 1c is a GC graph analyzing fragrance components according to Comparative Example 17 , FIG. 1D is a GC graph analyzing fragrance components according to Comparative Example 18, and FIG. 1E is a GC graph analyzing fragrance components according to Comparative Example 19.

2 is a GC graph in which fragrance components are analyzed according to Comparative Example 20.

The results of analyzing the fragrance components of Example 1 and Comparative Examples 16 to 19 measured in the GC graph are shown in Table 5 below.

division absorbent total peak area value Example 1 DVB/CAR/PDMS 470,117,445,946 Comparative Example 16 DVB/PDMS 288,965,928,582 Comparative Example 17 CAR/PDMS 395,286,579,041 Comparative Example 18 PDMS 279,857,742,683 Comparative Example 19 PA 130,432,768,228 Comparative Example 20 DVB/CAR/PDMS 171,727,219,832

As shown in Table 5 above, the fragrance components analyzed according to Example 1 of the present invention had a higher total peak area value than Comparative Examples 16 to 20, so it was confirmed that the content of fragrance components was high.

Claims (13)

(A) freeze-drying the citrus peel;
(B) pulverizing the lyophilized citrus rind and adsorbing and extracting the lyophilized rind through an adsorbent using a solid-phase microextraction arrow (SPME-Arrow) method; and
(C) putting the adsorbent into gas chromatography-mass spectrometry (GC-MS) and desorbing and analyzing volatile flavor components adsorbed and extracted by the adsorbent; A method for analyzing fragrance components. [Claim 2] The method for analyzing fragrance components of citrus peel according to claim 1, wherein the freeze-drying in step (A) is performed by freezing at -80 to -70 ° C for 15 to 25 hours. The method of claim 1, wherein the adsorbent in step (B) is divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS). The method of claim 1, wherein the extraction in step (B) is performed at an extraction temperature of 35 to 55 ° C, an extraction time of 50 to 70 minutes, and an equilibrium temperature of 5 to 25 minutes. method. According to claim 1, wherein the area value of the analyzed pericarp aroma component is 1.0X10 ¹¹ to 9.0X10 ¹¹ , characterized in that the method of analyzing the fragrance component of citrus peel. The method of claim 1, wherein the main aroma components of the analyzed citrus peel are β-Myrcene, D-Limonene, γ-Terpinene, p-cymene ( p-cymene), terpinolen, copaene, β-elemene, α-humulene, α-farnesene and linalo Aroma component analysis method of citrus peel, characterized in that it is linalool. The method of claim 1, wherein each area value of the major aroma component of the analyzed citrus peel is 1.0X10 ¹⁰ to 6.0X10 ¹⁰ of β-Myrcene, 1.0X10 ¹¹ to 5.0X10 ¹¹ of D-Limonene, 5.0X10 ¹⁰ to 9.9X10 ¹⁰ of γ-terpinene, 1.0X10 ¹⁰ to 5.0 X10 ¹⁰ of p-cymene (p-cymene), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Terpinolen (Terpinolen), 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of Copaene (Copaene), 1.0X10 ¹⁰ to 7.0X10 ¹⁰ of β-elemene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-Humulene, 1.0X10 ¹⁰ to 5.0X10 ¹⁰ of α-farnesene and 1.0X10 ¹⁰ to 5.0X10 ¹⁰ Linalool (Linalool) A method for analyzing the aroma component of citrus peel, characterized in that. The method of claim 7, wherein the total area value of the major aroma components of the analyzed citrus peel is 1.0X10 ¹¹ to 7.0X10 ¹¹ . The method of claim 1, wherein the citrus peel is citrus (Citrus unshiu), lemon (Citrus limon), orange (Citrus sinensis), citron (Citrus junos), grapefruit (Citrus paradisi) or hallabong (Citrus hybrid Shiranuhi) A method for analyzing the aroma components of citrus peels. The method of claim 1, wherein the analyzed fragrance component has an accuracy of 80% or more during validation. A perfume composition characterized in that it comprises a fragrance component of citrus peel analyzed by the analysis method according to any one of claims 1 to 10. A food composition characterized in that it contains a fragrance component of citrus peel analyzed by the analysis method of any one of claims 1 to 10. A cosmetic composition comprising a fragrance component of citrus peel analyzed by the analysis method of any one of claims 1 to 10.

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