CN109444284B - Method for simultaneously determining flavor substances in pepper and/or rattan pepper and oil products thereof - Google Patents
Method for simultaneously determining flavor substances in pepper and/or rattan pepper and oil products thereof Download PDFInfo
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Abstract
The invention provides a method for simultaneously measuring flavor substances in pepper and/or zanthoxylum schinifolium and oil products thereof, which is characterized by comprising the following steps of: the detection is carried out by adopting a high performance liquid chromatography, and the operation steps are as follows: 1) preparing a reference solution; 2) preparing a test solution; 3) respectively sucking the reference solution and the test solution and injecting into a liquid chromatograph. The method for simultaneously measuring the flavor substances in the pepper and/or rattan pepper and the oil products thereof can be used for simultaneously measuring the chemical components of the hemp, such as hydroxyl-alpha-sanshool, hydroxyl-beta-sanshool and hydroxyl-gamma-sanshool, and the chemical components of the incense, such as limonene and linalool, in the flavor substances in the pepper and/or rattan pepper and the oil products thereof. The method is simple and convenient, has reliable results, has important significance for the quality control and scientific evaluation of the zanthoxylum and/or zanthoxylum and related products thereof, and has good application prospect.
Description
Technical Field
The invention particularly relates to a method for simultaneously measuring flavor substances in pepper and/or zanthoxylum schinifolium and oil products thereof.
Background
The commercial pepper products mainly comprise two kinds of pepper (zanthoxylum bungeanum Maxim.) and zanthoxylum piperitum (zanthoxylum pipertum DC.), which are special medicinal and edible products. As an edible product, "numb and fragrant" is the main flavor characteristic, and is an indispensable condiment in Sichuan dishes with the characteristics of spicy, hot, fresh and fragrant; as a medicinal product, chemical components embodying the 'hemp and fragrance' are also main active components of the pepper, and researches show that the active components in pepper commodities have strong pharmacological activity on the aspects of digestion system, nervous system, circulatory system, inflammation resistance, pain relief, tumor resistance, oxidation resistance, bacteria resistance and insect killing. At present, the zanthoxylum bungeanum (rattan pepper) is mainly used as a food in two forms of a seasoning raw material and a processed product (zanthoxylum oil/rattan pepper oil), and the zanthoxylum oil (rattan pepper oil) is seasoning oil with numb fragrance prepared by directly soaking zanthoxylum bungeanum (rattan pepper) with edible oil or mixing zanthoxylum bungeanum (rattan pepper) essential oil prepared by using methods such as organic solvent extraction, supercritical CO2 extraction and the like with a proper amount of edible oil.
The fragrant substances in the pepper products are mainly volatile oil which comprises volatile components such as alkene, alcohol, ketone, aldehyde, ester and the like, the volatile oil in the pepper contains high content of limonene and linalool, wherein the content of limonene is the highest, and the rattan pepper contains high content of linalool and limonene, wherein the content of linalool is the highest; the numb taste substance is mainly chain unsaturated fatty acid amide substance represented by sanshool, mainly comprising hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, etc., wherein the content of the hydroxy-alpha-sanshool in the pepper and the zanthoxylum piperitum is highest. At present, in the quality evaluation standards of zanthoxylum bungeanum (rattan pepper) and zanthoxylum oil (rattan pepper oil), sensory evaluation is mostly taken as a main method, and the limit requirements of each standard have differences, so that the judgment of the spicy flavor of zanthoxylum bungeanum (rattan pepper) and zanthoxylum oil (rattan pepper oil) is lack of scientificity, has no accuracy and objectivity, and is difficult to realize standardization and quantification. Some studies have conducted quantitative analysis on the pungent substances or the fragrant substances, but the flavor substances of zanthoxylum bungeanum include the pungent substances and the fragrant substances. The single measurement of the content of the fragrant substances or the content of the numb substances as the quality evaluation standard of the zanthoxylum bungeanum or the zanthoxylum oil is not representative and cannot comprehensively and truly reflect the internal quality of the zanthoxylum bungeanum (zanthoxylum piperitum) and the zanthoxylum oil (zanthoxylum piperitum oil).
The existing method for detecting the content of the numb-taste substances mainly comprises an ultraviolet spectrophotometry method, a High Performance Liquid Chromatography (HPLC) method and the like, wherein the spectrophotometry method is used for detecting the content of the total numb-taste substances and cannot reflect the condition of each numb-taste component. The HPLC method has the characteristics of easy operation, high sensitivity, good stability and the like, and becomes a main method for analyzing and determining the spicy substances. However, in the measurement of the spicy substances in the zanthoxylum oil (zanthoxylum oil), methanol is often adopted to extract the spicy substances in the zanthoxylum oil (zanthoxylum oil), and two immiscible liquids formed after extraction are subjected to liquid separation and are analyzed by taking methanol liquid, so that incomplete extraction of the spicy substances in the zanthoxylum oil (zanthoxylum oil) is easily caused by the method, and the quantitative analysis is not accurate enough. At present, the content of the fragrance substances is usually determined by adopting a gas chromatography to determine the content of limonene and linalool, the gas chromatography is complex to operate, has higher risk coefficient and is difficult to popularize, the high performance liquid chromatography is adopted to determine the fragrance substances of pepper (zanthoxylum schinifolium) and pepper oil (zanthoxylum schinifolium oil) is rarely involved, and no report is provided for simultaneously determining the spicy substances and the fragrance substances. Therefore, a simple and accurate method for analyzing and determining the spicy substances and the fragrant substances in the zanthoxylum bungeanum (zanthoxylum piperitum) and the zanthoxylum oil (zanthoxylum piperitum oil) as common indexes is established, a scientific and objective quality standard is established for the method, and the method has important significance for the quality control and scientific evaluation of the zanthoxylum piperitum and related products thereof.
Disclosure of Invention
In order to solve the problems, the invention provides a method for simultaneously measuring flavor substances in pepper and/or zanthoxylum piperitum and oil products thereof, which is characterized by comprising the following steps: the detection is carried out by adopting a high performance liquid chromatography, and the operation steps are as follows:
1) preparation of control solutions: taking a hydroxy-alpha-sanshool, a hydroxy-beta-sanshool, a hydroxy-gamma-sanshool, limonene and linalool reference substance, adding isopropanol to dissolve, and freezing and storing to obtain a reference substance solution;
2) preparation of a test solution:
a, Chinese prickly ash or rattan pepper test products: pulverizing the sample, sieving, extracting with edible oil, filtering, dissolving the extractive solution in isopropanol, filtering, and collecting filtrate as sample solution;
b, testing pepper oil or rattan pepper oil: dissolving a sample in isopropanol, filtering, and taking the filtrate as a sample solution;
3) respectively sucking the reference solution and the test solution to be injected into a liquid chromatograph, wherein the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; mobile phase: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the gradient elution procedure was as follows:
further, the reference solution in the step 1) contains the concentrations of hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, limonene and linalool which are 800 mug/mL, 90 mug/mL, 80 mug/mL, 2000 mug/mL and 1000 mug/mL respectively.
Further, the freezing temperature in step 1) is-20 ℃.
Further, the screen mesh screened in the step a is a third screen mesh.
Further, the mass volume ratio of the to-be-detected sample of the zanthoxylum bungeanum or the zanthoxylum piperitum in the step a to the edible oil is 10g:60mL, and the edible oil is corn oil.
Further, the extraction in the step a is constant-temperature extraction, the extraction time is 150min, and the extraction temperature is 80 ℃.
Further, the volume ratio of the extracting solution in the step a to the isopropanol is 1: 50; and/or the volume ratio of the zanthoxylum oil or zanthoxylum oil sample to be detected to the isopropanol in the step b is 1: 50.
Further, the amounts of the reference solution and the sample solution to be injected into the liquid chromatograph in step 3) were 5. mu.l, respectively.
Further, the octadecylsilane chemically bonded silica chromatographic column in the step 3) is as follows: zishengtang CAPCELL PAK C18 column, 250mm × 4.6mm, 5 μm, MGII.
Further, the flow rate in the chromatographic conditions in the step 3) is 1.00 mL/min; the column temperature was 30 ℃.
Further, the wavelengths in the chromatographic conditions in the step 3) are respectively 200nm and 270 nm.
The method for simultaneously measuring the flavor substances in the pepper and/or rattan pepper and the oil products thereof can be used for simultaneously measuring the chemical components of the hemp, such as hydroxyl-alpha-sanshool, hydroxyl-beta-sanshool and hydroxyl-gamma-sanshool, and the chemical components of the incense, such as limonene and linalool, in the flavor substances in the pepper and/or rattan pepper and the oil products thereof. The method is simple and reliable in result, has important significance for quality control and scientific evaluation of pepper (rattan pepper) and related products, and has good application prospect.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 shows absorption spectra of hydroxy- α -sanshool, hydroxy- β -sanshool, and hydroxy- γ -sanshool at wavelength range of 190-400 nm.
FIG. 2 is the absorption spectrum of linalool and limonene within the wavelength range of 190-400 nm.
FIG. 3 gradient elution chromatogram of scheme 1 (A: wavelength 270nm, B: wavelength 200 nm).
FIG. 4 gradient elution chromatogram of scheme 2 (A: wavelength 270nm, B: wavelength 200 nm).
FIG. 5 gradient elution chromatogram of scheme 3 (A: wavelength 270nm, B: wavelength 200 nm).
FIG. 6 is a high performance liquid chromatogram of Zanthoxylum bungeanum (oil) (A: wavelength 270nm, B: wavelength 200 nm).
FIG. 7 is a high performance liquid chromatogram of Zanthoxylum piperitum (oil) (A: wavelength 270nm, B: wavelength 200 nm).
Detailed Description
1. Materials, reagents and apparatus
1.1 Experimental instruments
Shimadzu LC-2010A high performance liquid chromatograph, SIL-20A autosampler, SPD-20A detector, CTO-20A column oven, LC solution chromatography workstation software, Shimadzu corporation, Japan; TU-1901 double-beam UV-visible spectrophotometer (Beijing Pujingyo general instruments, Inc.); DZKW constant temperature water bath (Beijing Zhongxing Wei Co., Ltd.). SartoriusBP211D electronic balance (sartorius, germany); HX-200 type high-speed Chinese medicine grinder (hardware medical and medical apparatus works of Yangxi city of Yongkang, Zhejiang province); UPT-11-10T series super water purifier.
1.2 materials and reagents
Reference substance of numb-taste substance: the pepper spicy substance reference substance comprises: hydroxy-alpha-sanshool (self-made, purity 98.35%); hydroxyl-beta-sanshool (self-made, applied for national standard product, purity 98.27%); hydroxy-gamma-sanshool (self-made, purity 98.54%). Fragrance substance control: limonene (96% purity; lot No. 100472-201503) and linalool (97.8% purity; lot No. 111503-201603) controls were purchased from the Chinese food & drug testing institute. Isopropanol (analytically pure, metropolis chemical limited), methanol (chromatographically pure, sigma usa) acetonitrile (chromatographically pure, sigma usa), and water as ultrapure water self-made in the laboratory.
The pepper samples are collected from Sichuan Han source, Gansu Wudu and Shanxi Hancheng; collecting Zanthoxylum piperitum samples from Sichuan Santai, Sichuan Jinyang, and Sichuan Hongya; the zanthoxylum oil and zanthoxylum oil samples are purchased from supermarkets and comprise zanthoxylum oil in cattle market, zanthoxylum oil in nine-bucket shape, Yongfeng and zanthoxylum oil; wanfuteng pepper oil, Yaomazi rattan pepper oil and Nippon city rattan pepper oil.
Example 1 measurement of Pepper (Zanthoxylum schinifolium) flavor
1) Preparation of control solutions: precisely weighing hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, linalool and limonene reference substances in 5 volumetric flasks, dissolving the hydroxy-alpha-sanshool, the hydroxy-beta-sanshool, the linalool and the limonene reference substances respectively by using isopropanol, fixing the volume to a scale to obtain reference substance stock solutions with the concentrations of 8mg/mL, 0.9mg/mL, 0.8mg/mL, 20mg/mL and 10mg/mL, then respectively taking 1mL of mother solution in a 10mL volumetric flask, fixing the volume by using the isopropanol to prepare mixed reference substance mother solutions of 800 mu g/mL, 90 mu g/mL, 80 mu g/mL, 2000 mu g/mL and 1000 mu g/mL, and storing the mixed reference substance mother solutions at-20 ℃ for liquid phase analysis.
2) Preparation of a test solution:
crushing a zanthoxylum bungeanum (rattan pepper) sample into powder by a crusher (sieving by a third sieve), weighing 10g of the sample in a conical flask, adding 60mL of corn oil, leaching for 150min in a constant-temperature water bath kettle at 80 ℃, filtering to obtain a subsequent filtrate, sucking 1mL of zanthoxylum bungeanum extract oil (rattan pepper extract oil) in a 50mL volumetric flask by a liquid transfer gun, dissolving by isopropanol, fixing the volume to a scale mark, shaking uniformly, filtering by a 0.45 mu m microporous filter membrane, and obtaining the subsequent filtrate.
3) Respectively sucking 5 μ L of reference solution and test solution, and injecting into liquid chromatograph under the following chromatographic conditions:
a chromatographic column: zishengtang CAPCELL PAK C18Column (250 mm. times.4.6 mm, 5 μm; MGII); the column temperature is 30 ℃; mobile phase: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the flow rate of the mobile phase is 1.00 mL/min; the gradient elution procedure was as follows:
after the high performance liquid chromatography is adopted for detection, the contents of hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, linalool and limonene in the pepper (rattan pepper) sample are obtained through calculation and are shown in table 1.
TABLE 1 Zanthoxylum bungeanum (Zanthoxylum piperitum) flavor substance content
"-" indicates that the detection limit is exceeded
Example 2 measurement of Pepper oil (Zanthoxylum oil) flavor
1) Preparation of control solutions: precisely weighing hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, linalool and limonene reference substances into 5 volumetric flasks, dissolving the hydroxy-alpha-sanshool, the hydroxy-beta-sanshool, the linalool and the limonene reference substances respectively by using isopropanol, fixing the volume to a scale to obtain reference substance stock solutions with the concentrations of 8mg/mL, 0.9mg/mL, 0.8mg/mL, 20mg/mL and 10mg/mL, then respectively taking 1mL of mother solution into a 10mL volumetric flask, fixing the volume by using the isopropanol to prepare mixed reference substance solutions of 800 mu g/mL, 90 mu g/mL, 80 mu g/mL, 2000 mu g/mL and 1000 mu g/mL, and storing the mixed reference substance solutions at the temperature of-20 ℃ for liquid phase analysis.
2) Preparation of a test solution:
precisely sucking 1mL oleum Zanthoxyli Bungeani in a 50mL volumetric flask with a pipette, diluting to a constant volume with isopropanol to scale mark, shaking to dissolve oleum Zanthoxyli Bungeani (Zanthoxylum piperitum oil), filtering with 0.45 μm microporous membrane, and collecting filtrate;
3) respectively sucking 5 μ L of reference solution and test solution, and injecting into liquid chromatograph under the following chromatographic conditions:
a chromatographic column: zishengtang CAPCELL PAK C18Column (250 mm. times.4.6 mm, 5 μm; MGII); the column temperature is 30 ℃; mobile phase: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the flow rate of the mobile phase is 1.00 mL/min; the gradient elution procedure was as follows:
after the high performance liquid chromatography is adopted for detection, the contents of hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, limonene and linalool in the zanthoxylum oil sample are obtained by calculation and are shown in table 2.
TABLE 2 Zanthoxylum oil (Zanthoxylum oil) flavor substance content
The beneficial effects of the invention are illustrated by way of test examples below:
test example 1 chromatographic condition selection and methodological investigation
1 selection of detection wavelength
The maximum absorption wavelength was determined by scanning 3 kinds of sesame and 2 kinds of fragrance standard with an ultraviolet-visible spectrophotometer at full wavelength (in the range of 190-460 nm), and the results are shown in FIG. 1 and FIG. 2. From fig. 1, it can be seen that: the hydroxyl-alpha-sanshool, the hydroxyl-beta-sanshool and the hydroxyl-gamma-sanshool have maximum absorption peaks at 270 nm; from fig. 2, it follows that: linalool and limonene have maximum absorption peaks at 200 nm.
2 elution gradient program selection
Different elution gradient programs were set, and the protocols of the different elution programs are shown in tables 3, 4 and 5, and the high performance liquid chromatograms under the different protocols are shown in FIG. 3 (scheme 1), FIG. 4 (scheme 2) and FIG. 5 (scheme 3). The peak separation for each protocol is shown in Table 6.
Table 3 scheme 1 gradient elution procedure
Table 4 scheme 2 gradient elution procedure
Table 5 scheme 3 gradient elution procedure
TABLE 6 separation of flavors by different protocol gradient elution procedures
According to the peak shape and the separation degree of chromatographic peaks under each elution gradient program, the final determination is scheme 1: the mobile phase is acetonitrile-water, and the gradient elution condition is as follows: 0-15 min, acetonitrile-water (40: 60-50: 50); 15-25 min, acetonitrile-water (50: 50-60: 40); 25-35 min, acetonitrile-water (60: 40-70: 30); 35-60 min acetonitrile-water (70:30), column temperature 30 ℃, flow rate 1.0mL/min, sample volume 5 muL, detection wavelength 200nm, 270 nm. By adopting the gradient elution procedure, 3 main numb-flavor substances and two flavor substances can be completely separated within 60min, the separation degrees are all more than 1.5, the peak shapes are sharp, and the symmetry is good.
3 methodology examination
3.1 Linear relationship investigation
100 mul, 200 mul, 400 mul, 600 mul, 800 mul and 1000 mul of mother liquor of the mixed reference substance are respectively sucked by a pipette and put into 6 brown volumetric flasks of 2ml, and the volume is fixed to the scale mark by isopropanol to prepare standard solutions with different mass concentrations. The high performance liquid chromatogram of the sample is shown in figure 3, which is determined by injecting sample under the condition of item 2. Repeating the steps for 3 times, and performing linear regression by respectively taking the peak area as a vertical coordinate (Y) and the mass concentration (mu g/mL) as a horizontal coordinate (X), wherein the result shows that each spicy component and each fragrance component have a good linear relation with the peak area in each range, and the regression equation is detailed in the linear range and the related coefficient is detailed in Table 7.
TABLE 7 regression equation, Linear Range and correlation coefficient
3.2 precision test
Precisely absorbing 5 mu L of mixed reference substance solution, carrying out sample injection analysis under the condition of the item 2, carrying out sample injection for 6 times continuously, and measuring the peak areas, wherein RSD of the peak areas of the components are respectively 0.56% of hydroxyl-alpha-sanshool, 0.27% of hydroxyl-beta-sanshool, 0.38% of hydroxyl-gamma-sanshool, 0.76% of linalool and 0.68% of limonene, thus the precision of the instrument is good.
3.3 stability test
Precisely absorbing the same zanthoxylum oil sample solution, respectively injecting samples for 0, 4, 8, 12, 18 and 24 hours for determination according to the chromatographic condition under the item of 2, and determining that the peak areas RSD of the components are respectively 1.21 percent of hydroxyl-alpha-sanshool, 0.86 percent of hydroxyl-beta-sanshool, 0.75 percent of hydroxyl-gamma-sanshool, 0.98 percent of linalool and 1.07 percent of limonene, which indicates that the extracting solution is basically stable within 24 hours.
3.4 repeatability test
6 parts of test solution is prepared from the same brand of zanthoxylum oil by the method in the item of the example 3, and the RSD of the mass fractions of the components are respectively 0.77% of hydroxyl-alpha-sanshool, 0.61% of hydroxyl-beta-sanshool, 0.36% of hydroxyl-gamma-sanshool, 0.86% of linalool and 1.01% of limonene according to the determination analysis under the chromatographic condition of the item 2, which shows that the repeatability of the method is good.
3.5 sample application recovery test
Precisely sucking 0.5mL of 6 parts of zanthoxylum oil with known flavor substances of the same brand into a 50mL volumetric flask 1, respectively adding a proper amount of hydroxy-alpha-sanshool, hydroxy-beta-sanshool, hydroxy-gamma-sanshool, linalool and limonene standard substances, and then adding isopropanol to a constant volume. And the sample injection was carried out 6 times in the same manner as in example 2, and the peak areas were measured respectively to calculate the recovery rate and RSD value. The results are shown in Table 8.
TABLE 8 flavor ingredient recovery test results
3.6 content determination and verification of fructus Zanthoxyli (Zanthoxylum schinifolium L.) and oleum Zanthoxyli (Zanthoxylum schinifolium L.) fructus material and aromatic substance
According to the chromatographic conditions under item 2, 3 main spicy components and 2 main fragrance components in 12 samples of zanthoxylum bungeanum (Sichuan Hanyuan zanthoxylum bungeanum, Shaanxi Hancheng zanthoxylum bungeanum, Gansu Wudu zanthoxylum bungeanum), zanthoxylum piperitum (Sichuan three-platform zanthoxylum piperitum, Sichuan golden yang zanthoxylum piperitum, Sichuan hongya zanthoxylum piperitum), zanthoxylum oil (cattle slope zanthoxylum oil, Jiuguazhu zanthoxylum oil, Yongfeng and zanthoxylum oil), zanthoxylum oil (Yamao zanthoxylum oil, Wanfudajiao oil, cattle slope zanthoxylum oil) are simultaneously measured, the high performance liquid chromatogram of zanthoxylum piperitum (oil) is shown in fig. 6, the high performance liquid chromatogram result of zanthoxylum piperitum (oil) is shown in fig. 7, and the measurement results are the same as those in table 1 in example 1 and table 2 in example 2.
The results of chromatographic condition selection and methodology examination can be seen as follows: the detection method can be used for simultaneously detecting the chemical components of the hydroxyl-alpha-sanshool, the hydroxyl-beta-sanshool and the hydroxyl-gamma-sanshool of the hemp and the chemical components of the limonene and the linalool of the incense in the flavor substances of the zanthoxylum bungeanum (oil) and/or the zanthoxylum schinifolium (oil). The method has reliable results, has important significance for quality control and scientific evaluation of pepper (rattan pepper) and related products thereof, and has good application prospects.
Claims (10)
1. A method for simultaneously measuring flavor substances in pepper and/or rattan pepper and oil products thereof is characterized in that: the detection is carried out by adopting a high performance liquid chromatography, and the operation steps are as follows:
1) preparation of control solutions: taking a hydroxy-alpha-sanshool, a hydroxy-beta-sanshool, a hydroxy-gamma-sanshool, limonene and linalool reference substance, adding isopropanol to dissolve, and freezing and storing to obtain a reference substance solution;
2) preparation of a test solution:
a, Chinese prickly ash or rattan pepper test products: pulverizing the sample, sieving, extracting with edible oil, filtering, dissolving the extractive solution in isopropanol, filtering, and collecting filtrate as sample solution;
b, testing pepper oil or rattan pepper oil: dissolving a sample in isopropanol, filtering, and taking the filtrate as a sample solution;
3) respectively sucking the reference solution and the test solution to be injected into a liquid chromatograph, wherein the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; mobile phase: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the gradient elution procedure was as follows:
2. the method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the reference substance solution in the step 1) contains the concentrations of hydroxyl-alpha-sanshool, hydroxyl-beta-sanshool, hydroxyl-gamma-sanshool, limonene and linalool which are respectively 800 mu g/mL, 90 mu g/mL, 80 mu g/mL, 2000 mu g/mL and 1000 mu g/mL.
3. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the freezing temperature in step 1) is-20 ℃.
4. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the screen mesh for sieving in the step a is a third screen.
5. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the mass volume ratio of the sample to be detected of the zanthoxylum or the zanthoxylum schinifolium in the step a to the edible oil is 10g:60mL, and the edible oil is corn oil.
6. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the extraction in the step a is constant temperature extraction, the extraction time is 150min, and the extraction temperature is 80 ℃.
7. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the volume ratio of the extracting solution to the isopropanol in the step a is 1: 50; and (c) the volume ratio of the zanthoxylum oil or zanthoxylum oil to be detected to the isopropanol in the step b is 1: 50.
8. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the amount of the reference solution and the sample solution respectively injected into the liquid chromatograph in the step 3) is 5 mul.
9. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the octadecylsilane chemically bonded silica chromatographic column in the step 3) comprises the following steps: zishengtang CAPCELL PAK C18 column, 250mm × 4.6mm, 5 μm, MGII.
10. The method for simultaneously determining the flavor substances in the zanthoxylum bungeanum and/or the zanthoxylum piperitum and the oil products thereof according to claim 1, which is characterized in that: the flow rate in the chromatographic condition of the step 3) is 1.00ml/min, the column temperature is 30 ℃, and the wavelength is 200nm and 270 nm.
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| CN112683832B (en) * | 2020-12-18 | 2024-07-09 | 广电计量检测（成都）有限公司 | Detection method of spicy substances in fresh peppers |
| CN112684068A (en) * | 2020-12-21 | 2021-04-20 | 苏州华测生物技术有限公司 | Method for measuring content of hydroxy-alpha-sanshool |
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