CN113740447A - Identification and analysis method of key color generation substances of sorghum seed coats - Google Patents
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- 235000011684 Sorghum saccharatum Nutrition 0.000 title claims abstract description 72
- 239000000126 substance Substances 0.000 title claims abstract description 45
- 238000004458 analytical method Methods 0.000 title claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 22
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims abstract description 12
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 claims abstract description 12
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 235000009498 luteolin Nutrition 0.000 claims abstract description 12
- 238000001819 mass spectrum Methods 0.000 claims abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 40
- 239000000243 solution Substances 0.000 claims description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 10
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 238000012937 correction Methods 0.000 claims description 4
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- 229910021642 ultra pure water Inorganic materials 0.000 claims description 4
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- 238000004108 freeze drying Methods 0.000 claims description 2
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- 235000013339 cereals Nutrition 0.000 abstract description 5
- 239000012634 fragment Substances 0.000 abstract description 5
- 238000001228 spectrum Methods 0.000 abstract description 4
- 238000001514 detection method Methods 0.000 abstract description 3
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 3
- 238000002371 ultraviolet--visible spectrum Methods 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 abstract 1
- 150000002500 ions Chemical class 0.000 description 13
- 229920001864 tannin Polymers 0.000 description 7
- 239000001648 tannin Substances 0.000 description 7
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- 238000010521 absorption reaction Methods 0.000 description 6
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- 239000012528 membrane Substances 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 2
- 229930014669 anthocyanidin Natural products 0.000 description 2
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 2
- 235000008758 anthocyanidins Nutrition 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000004451 qualitative analysis Methods 0.000 description 2
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- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 2
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- 241000231392 Gymnosiphon Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001128140 Reseda Species 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- XADJWCRESPGUTB-UHFFFAOYSA-N apigenin Natural products C1=CC(O)=CC=C1C1=CC(=O)C2=CC(O)=C(O)C=C2O1 XADJWCRESPGUTB-UHFFFAOYSA-N 0.000 description 1
- 235000008714 apigenin Nutrition 0.000 description 1
- KZNIFHPLKGYRTM-UHFFFAOYSA-N apigenin Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 KZNIFHPLKGYRTM-UHFFFAOYSA-N 0.000 description 1
- 229940117893 apigenin Drugs 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229940030961 sorghum extract Drugs 0.000 description 1
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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- General Health & Medical Sciences (AREA)
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Abstract
The invention belongs to the field of analysis and detection, and particularly relates to a method for identifying and analyzing key color-producing substances of sorghum seed coats. The method specifically comprises the following steps: firstly, color-producing substances in sorghum seed coats are effectively extracted and separated, then key color-producing substances are determined by combining high performance liquid chromatography with ultraviolet-visible spectrum characteristics, finally fragment ion characteristics of the key color-producing substances are further determined by using an ultra-high performance liquid chromatography-time-of-flight mass spectrometer, and 2 key color-producing substances, namely luteolin and apiolin, are identified in sorghum by adopting the technical means of combining the spectra, the chromatography and the mass spectrum. The invention establishes a method for identifying and analyzing key color-producing substances in sorghum seed coats, can quickly and accurately identify compounds with main color-producing contributions in the sorghum seed coats, provides good technical guidance and data support for the research on the color quality of sorghum, is beneficial to establishing scientific sorghum grain color discrimination standards, and provides a new idea for the effective utilization of sorghum resources.
Description
Technical Field
The application belongs to the field of analysis and detection, and particularly relates to a method for identifying and analyzing key color generation substances of sorghum seed coats.
Background
The colors of the sorghum seed coats of different varieties are different, and the common sorghum seed coats have white, yellow, red or brown, black colors and the like. Traditionally, the color of sorghum seed coats is believed to be related to tannins, and all sorghum with red/brown seed coats contain tannin. Therefore, the seed coat color is an important evaluation basis in the use mode of the sorghum grains: when the sorghum is used as a feed, the digestive efficiency of animals is reduced due to the existence of tannin, so that the sorghum is mostly white or yellow seed coats; however, when the sorghum is used as a brewing raw material, the existence of tannin has a positive influence on brewing quality, and important aroma substances in wine bodies are generated after fermentation, so that red seed coat varieties are selected for brewing sorghum. However, modern researches show that the seed coat color and the color intensity are not completely reliable indexes of high and low tannin content in sorghum, and light-color sorghum grains also have a certain content of tannin; therefore, it is necessary to identify and analyze the color-producing substances in the sorghum seed coat to correctly distinguish two different properties, i.e., sorghum color and tannin content, and establish a scientific evaluation index of sorghum seed coat color. In literature reports, main chromogenic substances of plant seed coats are anthocyanidin and glycosides thereof, but no report exists at present on the relationship between the type of the anthocyanidin and the color of sorghum seed coats. At present, the color of the sorghum seed coat is judged mainly by artificial senses and lacks scientific quantitative indexes, so that the development of a method for identifying and analyzing key sorghum color development substances is beneficial to establishing a scientific sorghum color evaluation system, provides guidance for breeding sorghum with different purposes and promotes efficient utilization of sorghum germplasm resources.
Disclosure of Invention
In one aspect, the present application provides the use of luteolin and/or apidine for identifying the color of a sorghum seed coat.
In one aspect, the present application provides a method for identifying a color substance of sorghum seed coat, comprising the steps of:
s1, grinding the sorghum sample and screening the sorghum sample to prepare sorghum powder;
s2, adding glacial ethanol into the sorghum flour prepared in the step S1, and performing oscillation extraction;
s3, centrifuging after the extraction is finished in the step S2, and taking a supernatant V1;
s4, repeating the steps S2-S32 times to the residue obtained after the supernatant is obtained in the step S3, and obtaining supernatants V2 and V3;
s5, mixing the supernatants V1, V2 and V3, filtering for the first time, removing ethanol, freeze-drying, redissolving and filtering for the second time;
s6, carrying out HPLC-TOF-MS analysis on the sample prepared in the step S5.
In some embodiments, in step S1, the sorghum sample is ground and passed through a 0.5-3mm screen; in some embodiments, a 1mm screen is passed.
In some embodiments, in step S2, the mass-to-volume ratio of sorghum flour to glacial ethanol is: (1-5) g: (20-50) ml; in some embodiments, 1 g: 15 ml.
In some embodiments, the glacial ethanol is 60-90% glacial ethanol; in some embodiments, 80% glacial ethanol.
In some embodiments, the shaking extraction in step S3 is performed for 10-20 min; in some embodiments, the shaking table is shaking extracted for 15 min.
In some embodiments, centrifugation is performed at 2500-; in some embodiments, centrifugation is performed at 3000rpm for 10 min.
In some embodiments, in step S5, the first filtration is performed with a 0.45 μm filter; in some embodiments, ethanol is removed by rotary evaporation at 40-60 ℃; in some embodiments, ethanol is removed by rotary evaporation at 45 ℃; in some embodiments, the second filtration is with a 0.22 μm filter.
In some embodiments, in step S6, the liquid phase conditions are: a chromatographic column: ZORBAX C18 RP-HPLC column (250 mm. times.4.6 mm, 5 μm); mobile phase: 0.1% formic acid (dissolved in ultrapure water) (solution A), acetonitrile (solution B); flow rate: 0.5 mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; elution gradient: 5-15% of B (5min), 15-50% of B (40min), 50-70% of B (2min), 70-100% of B (1min), 100% of B (7min), 100-5% of B (1min) and 5% of B (9 min).
In some embodiments, the detector is a DAD detector.
In some embodiments, in step S6, the mass spectrometer uses Triple-TOF 5600+ time of flight liquid chromatography mass spectrometer, negative ion scan mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 50 psi; drying gas (GS 2): 50 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 550 ℃; ion source voltage (IS): -4500V; primary scanning: declustering voltage (DP): 80V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using a TOF MS-Product Ion-IDA mode, wherein CID energy is-20, -40 and-60V, and before sample injection, performing mass axis correction by using a CDS pump.
In some embodiments, the retention time of the chromogenic material is 18.23min and/or 20.84 min.
In some embodiments, the chromogenic material is luteolin and/or apidine.
In some embodiments, the luteolin is represented by formula I:
in some embodiments, the apidine is represented by formula II:
the method analyzes 2 key red-colored substances in the sorghum for the first time, and combines spectrum, chromatogram and mass spectrum technologies to identify two key color-producing substances, namely luteolin and apidine, in the sorghum, so that scientific basis and data support are provided for sorghum color evaluation, scientific sorghum color discrimination standard is favorably established, and meanwhile, reference ideas and methods are provided for identifying the color-producing substances in other plants.
Drawings
FIG. 1 is a liquid chromatogram of a sorghum extract at 473 nm;
FIG. 2 is a secondary mass spectrum of luteolin (Peak 1 of FIG. 1);
FIG. 3 is a secondary mass spectrum of apidine (peak 2 of FIG. 2);
FIG. 4 is a comparison of the maximum absorption wavelengths of reseda (left) and Peak 1 (right);
FIG. 5 is a comparison of maximum absorption wavelengths of apigenin standard (left) and peak 2 (right);
FIG. 6 shows the chemical structures of luteolin (left) and apiadine (right);
FIG. 7 is the color of the leach liquor of sample number 1 in Table 1;
FIG. 8 is the color of the leach liquor of sample number 2 in Table 1;
FIG. 9 is the color of the leach liquor of sample number 3 in Table 1.
Detailed Description
The technical solutions of the present invention are further illustrated by the following specific examples, which do not represent limitations to the scope of the present invention. Insubstantial modifications and adaptations of the present invention by others of the concepts fall within the scope of the invention.
Example 1
According to the method, firstly, color generation substances in sorghum seed coats are effectively extracted and separated, then key color generation substances are determined by combining high performance liquid chromatography with ultraviolet-visible spectrum characteristics, and finally fragment ion characteristics of the key color generation substances are further determined by an ultra high performance liquid chromatography-time-of-flight mass spectrometer, so that identification and analysis of the key color generation substances in sorghum are realized. The method comprises the following specific steps:
(1) color development substance extraction
Removing impurities from sorghum sample, grinding with a cyclone grinder, sieving with a 1mm sieve, weighing 2g sorghum powder, adding 30mL 80% glacial ethanol, and shaking and extracting for 15 min. After extraction, centrifugation is carried out for 10min at 3000 rpm. Removing supernatant, extracting residue, mixing the three supernatants, filtering with 0.45 μm filter membrane, rotary evaporating at 45 deg.C to remove ethanol, lyophilizing, redissolving, and filtering with 0.22 μm filter membrane for liquid sample injection.
(2) Liquid chromatography separation
And (3) separating main color-developing substances in the sorghum extracting solution by adopting an ultra-high performance liquid chromatography technology. Wherein:
the liquid phase conditions were: a chromatographic column: ZORBAX C18 RP-HPLC column (250 mm. times.4.6 mm, 5 μm); mobile phase: 0.1% formic acid (dissolved in ultrapure water) (solution A), acetonitrile (solution B); flow rate: 0.5 mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; elution gradient: 5-15% of B (5min), 15-50% of B (40min), 50-70% of B (2min), 70-100% of B (1min), 100% of B (7min), 100-5% of B (1min) and 5% of B (9 min).
The detector is a DAD detector, components with obvious absorption under 473nm are key red sorghum substances, and an ultraviolet visible light spectrum is obtained at the same time.
The mass spectrum adopts a Triple-TOF 5600+ flight time liquid mass spectrometer and a negative ion scanning mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 50 psi; drying gas (GS 2): 50 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 550 ℃; ion source voltage (IS): -4500V; primary scanning: declustering voltage (DP): 80V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using TOF MS-Product Ion-IDA modes, wherein CID energy is-20V, -40V and-60V, and before sample injection, performing mass axis correction by using a CDS (compact disc reader) pump to ensure that the error of the mass axis is less than 2 ppm.
(3) Qualitative analysis of key substances in red
And determining the main red contributing substances of the sorghum extracting solution by comparing the maximum absorption wavelength of a chromatographic peak, the molecular weight, the fragment ions of the secondary mass spectrum with the standard substance.
In this example, the retention time of the liquid chromatography of the key red substance was 18.23min, which was characterized as luteolin.
Example 2
According to the method, firstly, color generation substances in sorghum seed coats are effectively extracted and separated, then key color generation substances are determined by combining high performance liquid chromatography with ultraviolet-visible spectrum characteristics, and finally fragment ion characteristics of the key color generation substances are further determined by an ultra high performance liquid chromatography-time-of-flight mass spectrometer, so that identification and analysis of the key color generation substances in sorghum are realized. The method comprises the following specific steps:
(1) color development substance extraction
Removing impurities from sorghum sample, grinding with a cyclone grinder, sieving with a 1mm sieve, weighing 2g sorghum powder, adding 30mL 80% glacial ethanol, and shaking and extracting for 15 min. After extraction, centrifugation is carried out for 10min at 3000 rpm. Removing supernatant, extracting residue, mixing the three supernatants, filtering with 0.45 μm filter membrane, rotary evaporating at 45 deg.C to remove ethanol, lyophilizing, redissolving, and filtering with 0.22 μm filter membrane for liquid sample injection.
(2) Liquid chromatography separation
And (3) separating main color-developing substances in the sorghum extracting solution by adopting an ultra-high performance liquid chromatography technology. Wherein:
the liquid phase conditions were: a chromatographic column: ZORBAX C18 RP-HPLC column (250 mm. times.4.6 mm, 5 μm); mobile phase: 0.1% formic acid (dissolved in ultrapure water) (solution A), acetonitrile (solution B); flow rate: 0.5 mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; elution gradient: 5-15% of B (5min), 15-50% of B (40min), 50-70% of B (2min), 70-100% of B (1min), 100% of B (7min), 100-5% of B (1min) and 5% of B (9 min).
The detector is a DAD detector, components with obvious absorption under 473nm are key red sorghum substances, and an ultraviolet visible light spectrum is obtained at the same time.
The mass spectrum adopts a Triple-TOF 5600+ flight time liquid mass spectrometer and a negative ion scanning mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 50 psi; drying gas (GS 2): 50 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 550 ℃; ion source voltage (IS): -4500V; primary scanning: declustering voltage (DP): 80V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using TOF MS-Product Ion-IDA modes, wherein CID energy is-20V, -40V and-60V, and before sample injection, performing mass axis correction by using a CDS (compact disc reader) pump to ensure that the error of the mass axis is less than 2 ppm.
(3) Qualitative analysis of key substances in red
And determining the main red contributing substances of the sorghum extracting solution by comparing the maximum absorption wavelength of a chromatographic peak, the molecular weight, the fragment ions of the secondary mass spectrum with the standard substance.
In this example, the retention time of the liquid chromatography of the key red substance was 20.84min, which was characterized as apidine.
Example 3
In the embodiment, three sorghum samples with different skin reds and shades are selected, the extracting solution is obtained by processing the sorghum samples with the same method as the embodiment 1, and then the delta a value of the extracting solution relative to a reference white board is analyzed by a color difference meter; the content of luteolin and apidine in the sorghum grains is quantified by adopting the liquid chromatography technology of the invention and converted into the content (mg/100g) in the sorghum grains. Wherein, when the Delta a is a positive value, the red is shown to be deviated, and the larger the value is, the deeper the red is shown, and the color of the leaching liquor has good correlation with the color of the sorghum seed coat.
The results in table 1 show that the content of the key red-colored substances of the sorghum seed coat identified by the invention has good correlation with the color of the sample leaching liquor, so that the identification and analysis method of the key color-developing substances of the sorghum seed coat established by the invention has good applicability.
TABLE 1 analysis results of color and content of color-forming substances of three sorghum samples
Note: n.d represents no detection.
Claims (10)
1. Application of luteolin and/or apiadine in identifying sorghum seed coat color is provided.
2. A method for identifying a color substance of sorghum seed coats is characterized by comprising the following steps:
s1, grinding the sorghum sample and screening the sorghum sample to prepare sorghum powder;
s2, adding glacial ethanol into the sorghum flour prepared in the step S1, and performing oscillation extraction;
s3, centrifuging after the extraction is finished in the step S2, and taking a supernatant V1;
s4, repeating the steps S2-S32 times to the residue obtained after the supernatant is obtained in the step S3, and obtaining supernatants V2 and V3;
s5, mixing the supernatants V1, V2 and V3, filtering for the first time, removing ethanol, freeze-drying, redissolving and filtering for the second time;
s6, carrying out HPLC-TOF-MS analysis on the sample prepared in the step S5.
3. The method of claim 2, wherein in step S1, the sorghum sample is ground and passed through a 0.5-3mm sieve;
preferably, a 1mm screen.
4. The method of claim 2, wherein in the step S2, the mass-to-volume ratio of the sorghum flour to the glacial ethanol is: (1-5) g: (20-50) ml; preferably, 1 g: 15 ml;
preferably, the glacial ethanol is 60-90% glacial ethanol; preferably, 80% of glacial ethanol.
5. The method of claim 2, wherein the shaking extraction is performed for 10-20min in step S3; preferably, shaking table extracting for 15 min;
preferably, centrifugation is carried out for 5-15min at the rotation speed of 2500-;
preferably, centrifugation is carried out for 10min at 3000 rpm.
6. The method of claim 2, wherein in step S5, the first filtration is performed with a 0.45 μm filter;
preferably, the ethanol is removed by rotary evaporation at 40-60 ℃; preferably, ethanol is removed by rotary evaporation at 45 ℃;
preferably, the second filtration is performed with a 0.22 μm filter.
7. The method of claim 2, wherein in step S6, the liquid phase conditions are: a chromatographic column: ZORBAX C18 RP-HPLC column (250 mm. times.4.6 mm, 5 μm); mobile phase: 0.1% formic acid (dissolved in ultrapure water) (solution A), acetonitrile (solution B); flow rate: 0.5 mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; elution gradient: 5% -15% of B (5min), 15% -50% of B (40min), 50% -70% of B (2min), 70% -100% of B (1min), 100% of B (7min), 100% -5% of B (1min) and 5% of B (9 min);
preferably, the detector is a DAD detector.
8. The method of claim 2, wherein in step S6, the mass spectrometer is in Triple-TOF 5600+ time-of-flight hplc, negative ion scan mode; scanning range: m/z 100-1500; atomizing gas (GS 1): 50 psi; drying gas (GS 2): 50 psi; air curtain gas (CUR): 35 psi; ion source Temperature (TEM): 550 ℃; ion source voltage (IS): -4500V; primary scanning: declustering voltage (DP): 80V; focus voltage (CE): 10V; secondary scanning: and (3) acquiring mass spectrum data by using a TOF MS-Product Ion-IDA mode, wherein CID energy is-20, -40 and-60V, and before sample injection, performing mass axis correction by using a CDS pump.
9. The method according to any one of claims 2 to 8, wherein the retention time of the chromogenic substance is 18.23min and/or 20.84 min;
preferably, the coloring matter is luteolin and/or apiadine.
10. The use of claim 1 or the method of claim 9, wherein the luteolin is of formula I:
preferably, the apidine is represented by formula II:
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101355995A (en) * | 2005-10-03 | 2009-01-28 | 玛尔斯有限公司 | Improved process for analyzing for separating, and for isolating polar protic monomers and/or oligomers |
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2021
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