CN117420256B - Method for detecting anthraquinone compounds and application thereof - Google Patents
Method for detecting anthraquinone compounds and application thereof Download PDFInfo
- Publication number
- CN117420256B CN117420256B CN202311335576.9A CN202311335576A CN117420256B CN 117420256 B CN117420256 B CN 117420256B CN 202311335576 A CN202311335576 A CN 202311335576A CN 117420256 B CN117420256 B CN 117420256B
- Authority
- CN
- China
- Prior art keywords
- anthraquinone
- anthraquinone compounds
- paradibenzoic
- sample
- dihydrazide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 title claims abstract description 179
- -1 anthraquinone compounds Chemical class 0.000 title claims abstract description 162
- 238000000034 method Methods 0.000 title claims abstract description 45
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 122
- 238000001556 precipitation Methods 0.000 claims abstract description 39
- 238000001514 detection method Methods 0.000 claims abstract description 33
- 238000012216 screening Methods 0.000 claims abstract description 15
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 claims abstract description 10
- 239000002253 acid Substances 0.000 claims description 54
- 241000233866 Fungi Species 0.000 claims description 52
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 48
- 238000000855 fermentation Methods 0.000 claims description 48
- 230000004151 fermentation Effects 0.000 claims description 48
- 239000002904 solvent Substances 0.000 claims description 41
- 150000004056 anthraquinones Chemical class 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 16
- LLZYZGMVXFGQAY-UHFFFAOYSA-N 1,5-dihydroxy-3-methoxy-7-methylanthracene-9,10-dione Chemical compound CC1=CC(O)=C2C(=O)C3=CC(OC)=CC(O)=C3C(=O)C2=C1 LLZYZGMVXFGQAY-UHFFFAOYSA-N 0.000 claims description 14
- BTLXPCBPYBNQNR-UHFFFAOYSA-N 1-hydroxyanthraquinone Chemical compound O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2O BTLXPCBPYBNQNR-UHFFFAOYSA-N 0.000 claims description 14
- 239000006228 supernatant Substances 0.000 claims description 14
- 238000004885 tandem mass spectrometry Methods 0.000 claims description 8
- 230000003197 catalytic effect Effects 0.000 claims description 7
- 238000004817 gas chromatography Methods 0.000 claims description 7
- RHMXXJGYXNZAPX-UHFFFAOYSA-N emodin Chemical compound C1=C(O)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O RHMXXJGYXNZAPX-UHFFFAOYSA-N 0.000 claims description 6
- VWBVCOPVKXNMMZ-UHFFFAOYSA-N 1,5-diaminoanthracene-9,10-dione Chemical compound O=C1C2=C(N)C=CC=C2C(=O)C2=C1C=CC=C2N VWBVCOPVKXNMMZ-UHFFFAOYSA-N 0.000 claims description 5
- KHUFHLFHOQVFGB-UHFFFAOYSA-N 1-aminoanthracene-9,10-dione Chemical compound O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2N KHUFHLFHOQVFGB-UHFFFAOYSA-N 0.000 claims description 5
- BOCJQSFSGAZAPQ-UHFFFAOYSA-N 1-chloroanthracene-9,10-dione Chemical compound O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2Cl BOCJQSFSGAZAPQ-UHFFFAOYSA-N 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- 238000004448 titration Methods 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 239000002244 precipitate Substances 0.000 abstract description 54
- 238000006555 catalytic reaction Methods 0.000 abstract description 4
- 125000003277 amino group Chemical group 0.000 abstract description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 90
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 60
- 238000006243 chemical reaction Methods 0.000 description 16
- 230000001580 bacterial effect Effects 0.000 description 10
- 235000010633 broth Nutrition 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 8
- FRASJONUBLZVQX-UHFFFAOYSA-N 1,4-naphthoquinone Chemical compound C1=CC=C2C(=O)C=CC(=O)C2=C1 FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 6
- 239000007789 gas Substances 0.000 description 6
- 230000006872 improvement Effects 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 5
- 239000003086 colorant Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- YYVYAPXYZVYDHN-UHFFFAOYSA-N 9,10-phenanthroquinone Chemical compound C1=CC=C2C(=O)C(=O)C3=CC=CC=C3C2=C1 YYVYAPXYZVYDHN-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- YDQWDHRMZQUTBA-UHFFFAOYSA-N Aloe emodin Chemical compound C1=CC=C2C(=O)C3=CC(CO)=CC(O)=C3C(=O)C2=C1O YDQWDHRMZQUTBA-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- LQGUBLBATBMXHT-UHFFFAOYSA-N chrysophanol Chemical compound C1=CC=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O LQGUBLBATBMXHT-UHFFFAOYSA-N 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- FFWOKTFYGVYKIR-UHFFFAOYSA-N physcion Chemical compound C1=C(C)C=C2C(=O)C3=CC(OC)=CC(O)=C3C(=O)C2=C1O FFWOKTFYGVYKIR-UHFFFAOYSA-N 0.000 description 2
- PKUBGLYEOAJPEG-UHFFFAOYSA-N physcion Natural products C1=C(C)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O PKUBGLYEOAJPEG-UHFFFAOYSA-N 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- RZVHIXYEVGDQDX-UHFFFAOYSA-N 9,10-anthraquinone Chemical group C1=CC=C2C(=O)C3=CC=CC=C3C(=O)C2=C1 RZVHIXYEVGDQDX-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 1
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 description 1
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 description 1
- VWDXGKUTGQJJHJ-UHFFFAOYSA-N Catenarin Natural products C1=C(O)C=C2C(=O)C3=C(O)C(C)=CC(O)=C3C(=O)C2=C1O VWDXGKUTGQJJHJ-UHFFFAOYSA-N 0.000 description 1
- 239000010282 Emodin Substances 0.000 description 1
- RBLJKYCRSCQLRP-UHFFFAOYSA-N Emodin-dianthron Natural products O=C1C2=CC(C)=CC(O)=C2C(=O)C2=C1CC(=O)C=C2O RBLJKYCRSCQLRP-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- YOOXNSPYGCZLAX-UHFFFAOYSA-N Helminthosporin Natural products C1=CC(O)=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O YOOXNSPYGCZLAX-UHFFFAOYSA-N 0.000 description 1
- UGNZSMZSJYOGNX-UHFFFAOYSA-N Isoviocristine Natural products O=C1C=C(C)C(=O)C2=CC3=CC(OC)=CC(O)=C3C(O)=C21 UGNZSMZSJYOGNX-UHFFFAOYSA-N 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- NTGIIKCGBNGQAR-UHFFFAOYSA-N Rheoemodin Natural products C1=C(O)C=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1O NTGIIKCGBNGQAR-UHFFFAOYSA-N 0.000 description 1
- 240000004980 Rheum officinale Species 0.000 description 1
- 235000008081 Rheum officinale Nutrition 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- WLXGUTUUWXVZNM-UHFFFAOYSA-N anthraglycoside A Natural products C1=C(C)C=C2C(=O)C3=CC(OC)=CC(O)=C3C(=O)C2=C1OC1OC(CO)C(O)C(O)C1O WLXGUTUUWXVZNM-UHFFFAOYSA-N 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- NZPQWZZXRKZCDU-UHFFFAOYSA-N chrysophanol Natural products Cc1cc(O)c2C(=O)c3c(O)cccc3Oc2c1 NZPQWZZXRKZCDU-UHFFFAOYSA-N 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- VASFLQKDXBAWEL-UHFFFAOYSA-N emodin Natural products OC1=C(OC2=C(C=CC(=C2C1=O)O)O)C1=CC=C(C=C1)O VASFLQKDXBAWEL-UHFFFAOYSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 150000004338 hydroxy anthraquinones Chemical class 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000008141 laxative Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001543 purgative effect Effects 0.000 description 1
- 238000013102 re-test Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- G01N31/02—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using precipitation
-
- 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/025—Gas chromatography
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The application relates to the technical field of anthraquinone compound detection, and particularly discloses a method for detecting anthraquinone compounds and application thereof. The detection method comprises the following steps: sulfuric acid and paradibenzoic dihydrazide are added into a sample to be detected, and if purple precipitation is generated, the sample is primarily judged to contain the anthraquinone compounds. Anthraquinone compounds have two carbonyl groups oppositely positioned on the same six-carbon ring, and p-dibenzoic dihydrazide has two hydrazide groups oppositely positioned on the same benzene ring. If the sample contains anthraquinone compounds with a certain concentration, under the catalysis of sulfuric acid, the amino groups on the two opposite hydrazide groups and the two opposite carbonyl groups are complexed to form purple precipitates, and the more the generated purple precipitates are, the darker the color of the precipitates is. The method for detecting the anthraquinone compounds can be used for screening samples containing the anthraquinone compounds with a certain concentration or above.
Description
Technical Field
The application relates to the technical field of anthraquinone compound detection, in particular to a method for detecting anthraquinone compounds and application thereof.
Background
Some metabolites of fungi contain anthraquinone compounds. Anthraquinone compounds belong to polycyclic aromatic hydrocarbon compounds, have a multi-benzene ring conjugated system, have high stability and are widely applied to the industry of dye preparation. Some anthraquinone compounds also have hemostatic, antibacterial, purgative, and diuretic effects.
The detection of anthraquinone compounds is generally carried out directly by using gas chromatography, liquid chromatography, mass spectrometry and the like, when a plurality of samples need to be detected, for example, tens of fungus fermentation broths are detected, so that when fungus with high yield of anthraquinone compounds is screened, the detection is carried out directly by using the instruments, so that the time is consumed, and the detection cost is high.
For identifying anthraquinone compounds, such as hydroxy anthraquinone, alkali may be added to the sample to make the sample dark, and if the sample has red or purplish red color, the sample has light color after adding acid, and the sample has dark color after adding alkali, and the sample has red or purplish red color, and the sample is determined to contain hydroxy anthraquinone. However, the identification method can identify fewer anthraquinone types, and generally has an identification effect on hydroxyanthraquinone compounds.
Disclosure of Invention
In order to improve the conditions that the detection of anthraquinone compounds is generally carried out directly by using gas chromatography, liquid chromatography, mass spectrometry and the like, when a plurality of samples need to be detected, for example, detection of the anthraquinone compounds is carried out on dozens of fungus fermentation broths so as to screen fungus with high yield of anthraquinone compounds, the detection is carried out directly by using the instruments for one-to-one time consumption, and the detection cost is high, the application provides a method for detecting the anthraquinone compounds and application thereof.
In a first aspect, the present application provides a method for detecting anthraquinone compounds, and adopts the following technical scheme.
A method of detecting an anthraquinone compound, the detection method comprising: the anthraquinone compound has two carbonyl groups which are relatively aligned on the same six-carbon ring, and the carbon of the two carbonyl groups is derived from the six-carbon ring. Sulfuric acid and paradibenzoic dihydrazide are added into a sample to be detected, and if purple precipitation is generated, the sample is primarily judged to contain the anthraquinone compounds.
By adopting the technical scheme, the anthraquinone compound has two carbonyl groups which are oppositely positioned on the same six-carbon ring, and the paradibenzoic acid dihydrazide has two hydrazide groups which are oppositely positioned on the same benzene ring. If the sample contains anthraquinone compounds with a certain concentration, under the catalysis of sulfuric acid, the amino groups on the two opposite hydrazide groups and the two opposite carbonyl groups are complexed to form purple precipitates, and the more the generated purple precipitates are, the darker the color of the precipitates is. If no purple precipitate is generated: the sample may be free of the anthraquinone compound; it is also possible to have a lower concentration of the anthraquinone compound, for example, an anthraquinone compound mass concentration of 0.001% or less; it is also possible that the concentration of the anthraquinone compound is high, for example, the anthraquinone compound is 0.01% by mass or more, but the amount of the paradibenzoic dihydrazide added is too small, so that the reaction product is small, and the precipitation phenomenon cannot be observed. In the process of implementing the embodiment, when the mass concentration of the anthraquinone compound in the sample reaches 0.01%, if excessive paradibenzoic dihydrazide is added, for example, when the mass concentration of the paradibenzoic dihydrazide reaches 0.01% or more after the paradibenzoic dihydrazide is added into the sample, the anthraquinone compound is completely reacted, and the purple precipitation phenomenon is obvious. The method for detecting the anthraquinone compounds can be used for screening samples containing the anthraquinone compounds with a certain concentration or more, for example, samples with the anthraquinone compound mass concentration of more than 0.01% can be detected. Since the more violet precipitates are produced, the darker the precipitates are, and the method for detecting anthraquinone compounds can also be used for screening one or a plurality of samples with the darkest precipitates.
As an improvement of the method for detecting anthraquinone compounds, after preliminary determination that the sample contains the anthraquinone compounds, the type and concentration of anthraquinone compounds in the sample are also tested using gas chromatography-tandem mass spectrometry.
By adopting the technical scheme, after the anthraquinone compounds are primarily determined according to the purple precipitation phenomenon, in order to improve the accuracy of the determination result and detect the types and the concentrations of the anthraquinone compounds, the gas chromatography-tandem mass spectrometry is also used for retesting, and the gas chromatography-tandem mass spectrometry is reliable in detection result and can further verify the accuracy of the primary determination.
As an improvement of the method for detecting anthraquinone compounds, the sulfuric acid and the paradibenzoic dihydrazide are added into a sample to be detected at the temperature of 50-60 ℃ during detection.
By adopting the technical scheme, in the temperature of 50-60 ℃, compared with the normal temperature environment of 20-30 ℃, the reaction of the anthraquinone compound, sulfuric acid and paradibenzoic acid dihydrazide is faster.
As an improvement of the method for detecting anthraquinone compounds, the anthraquinone compounds are one or more of anthraquinone, 1-hydroxy anthraquinone, 1-chloro anthraquinone, 1-amino anthraquinone, 1, 5-diamino anthraquinone, 1,3, 8-trihydroxy-6-methyl anthraquinone and 1, 5-dihydroxyl-3-methoxy-7-methyl anthraquinone.
By adopting the technical scheme, the purple settlement phenomenon of the anthraquinone compounds after the reaction with sulfuric acid and paradibenzoic acid dihydrazide is obvious and easy to identify.
As an improvement of the method for detecting anthraquinone compounds, the sample is as follows: the sulfuric acid: the p-dibenzoic dihydrazide=1: (0.01-0.2): (0.001-0.03).
By adopting the technical scheme, if the anthraquinone compounds in the sample with the concentration of 0.01% of anthraquinone compounds completely react, the purple precipitation phenomenon is obvious, and for the anthraquinone compounds with the concentration of 0.01%, the sulfuric acid only needs to catalyze the amount, and the paradibenzoic acid dihydrazide with the proportion is in an excessive state, so that the sample with the mass concentration of the anthraquinone compounds reaching more than 0.01% can be detected. The anthraquinone compound and the paradibenzoic dihydrazide are complexed in a molar ratio of 1:1. The molar mass of the paradibenzoic acid dihydrazide is 194g/mol, the molar mass of the anthraquinone is 208g/mol, and the anthraquinone is the substance with the minimum molar mass in anthraquinone compounds, so that the molar mass of the paradibenzoic acid dihydrazide is slightly smaller than that of the anthraquinone and is also smaller than that of any other anthraquinone. For compounds of equal mass, the smaller the molar mass, the greater the molar mass. If the mass of the added paradibenzoic acid dihydrazide is not less than that of the anthraquinone compound, the paradibenzoic acid dihydrazide is in an excessive state, and the anthraquinone compound in the sample can be basically reacted completely. For mass ratio, samples: sulfuric acid: p-dibenzoic dihydrazide=1: (0.01-0.2): (0.001 to 0.03), the amount of sulfuric acid required is excessive, and if the minimum amount of 0.001 parts by mass of paradibenzoic acid dihydrazide is added, at least 0.001 parts by mass of anthraquinone compound can be reacted, so that 0.001 parts by mass of paradibenzoic acid dihydrazide is excessive relative to 0.01% (0.0001 parts by mass) of anthraquinone compound, that is, a sample having an anthraquinone compound mass concentration of 0.01% or more can be detected by adding the mixture. The ratio of the anthraquinone compounds produced by the fungus is generally less than 1mg/g, namely the mass of the anthraquinone compounds produced by each g of fungus fermentation liquor is less than 1mg, generally 0-700 mug, so that samples are selected: sulfuric acid: p-dibenzoic dihydrazide=1: (0.01-0.2): (0.001-0.03), the paradibenzoic dihydrazide is in an excessive state, and the anthraquinone compound in the sample can be completely reacted. For mass ratio, samples: p-dibenzoic dihydrazide=1: (0.001 to 0.03), the content of the paradibenzoic acid dihydrazide may be 0.001, 0.005, 0.01, 0.02 or 0.03, etc., and the content of the paradibenzoic acid dihydrazide may be not limited thereto, but may be 0.03 or more, for example, 0.04 to 0.1, etc.
In a second aspect, the application also provides an application of the method for detecting anthraquinone compounds, and adopts the following technical scheme.
The application of the method for detecting the anthraquinone compounds is that the method for detecting the anthraquinone compounds is applied to screening fungus and fermentation conditions of relatively high-yield anthraquinone compounds; the sample is taken from fungus fermentation liquor; and adding a catalytic amount of sulfuric acid and an excessive amount of paradibenzoic dihydrazide into the sample, and screening out fungus corresponding to one or more samples with the highest precipitation color and fermentation conditions of the fungus.
By adopting the technical scheme, the method for detecting the anthraquinone compounds is utilized for various fungus fermentation broths, and excessive paradibenzoic dihydrazide is added into the fungus fermentation broths, so that the darker the generated purple precipitate color is, the thicker the anthraquinone compounds contained in the fungus fermentation broths are. The catalytic amount of sulfuric acid may be 0.01 to 0.2% by volume of the sample, or may be 0.01% or less, for example, 0.001%. The fungus liquid sample with the same volume is taken, and fungus which can produce anthraquinone compounds relatively high and fermentation conditions thereof can be identified by adopting the detection method, and the fungus can be screened out for further culture fermentation. The relatively high yield, i.e. the one with the highest yield, or the first few with the highest yield, may also be one to more of the first few with the highest yield. The fungus and fermentation conditions of the relatively high-yield anthraquinone compounds correspond to one or more samples which generate purple sediment and have the deepest sediment color, for example, the sample with the mass concentration of the anthraquinone compounds in fungus fermentation liquor of more than or equal to 0.01 percent, the sample with the highest yield of the anthraquinone compounds in the same fungus strain and different partial fermentation conditions, the sample with the highest yield of the anthraquinone compounds in a plurality of fungus strains and a plurality of fermentation conditions, or the first samples with the highest yield, or one to more samples with the first samples with the highest yield. In the application, the type and concentration of anthraquinone compounds in the sample can be retested by using a gas chromatography tandem mass spectrometry after precipitation detection. The detection method can reflect the relative concentration of the anthraquinone compounds in the sample, can be used as an important reference factor for screening fungus and culturing conditions, and can comprehensively consider factors such as cost to screen proper fungus and fermenting conditions thereof during specific selection. One or more samples with the deepest precipitation color can be one sample, two samples or three samples with the deepest precipitation color in a plurality of samples, one or more samples with the deepest precipitation color in the first few samples, or all samples with the anthraquinone compound mass concentration of more than or equal to 0.01 percent, the samples with the anthraquinone compound mass concentration of less than or equal to 0.01 percent can be used for carrying out precipitation reaction, the precipitation color of the samples is recorded by photographing, a colorimetric card is manufactured, and the retest is reserved when the precipitation color of the tested samples is more than or equal to the precipitation color of the colorimetric card. In the comparison of the precipitation colors, excessive paradibenzoic dihydrazide is added to make the anthraquinone compounds fully react.
As an improvement of the application of the method for detecting anthraquinone compounds, before adding sulfuric acid and paradibenzoic dihydrazide, fungus crushing and centrifuging are carried out on the fungus fermentation liquor to obtain supernatant, the obtained supernatant is directly used as the sample to be detected, or the obtained supernatant is extracted and then the extract is used as the sample to be detected.
By adopting the technical scheme, the treated bacterial liquid is clear, and if the bacterial liquid contains anthraquinone compounds, the bacterial liquid can fully react with sulfuric acid and paradibenzoic dihydrazide, so that the reaction phenomenon can be observed more easily. Alternatively, the treated bacterial liquid can be extracted, the anthraquinone compound solution can be concentrated by using an extracting solution with the volume smaller than that of the supernatant, then the extracting solution is detected, the precipitation color is darker during the reaction, the phenomenon is more obvious, and the observation is more convenient. The solvent used for extraction can be dichloromethane, or dichloromethane with the volume ratio of 7:3: other suitable solvents may be used as the acetone mixed solvent. The supernatant obtained may also be filtered, for example, through a filter membrane having a pore size of 0.22. Mu.m, to obtain a sample to be tested. In the application process, whether the reactant generates sediment and the color depth of the sediment reflect the concentration of the anthraquinone compounds in the current sample can be used as an important reference factor for screening fungus with relatively high yield of anthraquinone compounds and the fermentation condition of the fungus. In order to more accurately screen fungus with high yield of anthraquinone compounds, the same and sufficient fermentation time is preferably set, so that the fungus is fully fermented, and the screening accuracy is improved.
As an improvement of the application of the method for detecting anthraquinone compounds, adopting a titration mode, wherein the supernatant is directly used as the sample to be detected, and the paradibenzoic acid dihydrazide added into the sample to be detected is added in the form of paradibenzoic acid dihydrazide solution with the concentration of 0.04-0.1 g/mL; the solvent of the paradibenzoic acid dihydrazide solution is ethanol; by volume, the sample: the paradibenzoic dihydrazide solution = 1: (0.03-0.20).
By adopting the technical scheme, as the paradibenzoic dihydrazide is easy to dissolve in the ethanol, and the compatibility of the ethanol and the sulfuric acid is high, the catalysis and the complexation reaction of the sulfuric acid are facilitated. The concentration of the anthraquinone compound in the supernatant is generally less than 0.0007g/mL of the paradibenzoic dihydrazide solution with the concentration of 0.04-0.1 g/mL. The density of the supernatant is basically equal to that of water, and if the volume of the sample is 1mL, the mass of the anthraquinone compound is generally not more than 0.0007g; correspondingly, the minimum volume of the dibenzoic dihydrazide solution is 0.03mL, and the mass of the paradibenzoic dihydrazide reaches 0.0012g, so that the anthraquinone compound can be completely reacted. The method is characterized in that a plurality of samples are detected, a solution turbidity phenomenon is generated during precipitation, namely floccules are scattered in a reaction solution, and then the floccules are redeposited, so that the precipitation phenomenon can be obviously observed, and fungus with high-yield anthraquinone compounds and fermentation conditions of the fungus can be screened out according to the difference of precipitation colors (turbidity color depths).
In summary, the method for detecting anthraquinone compounds and application thereof have the following beneficial effects:
In the detection method, the anthraquinone compound has two carbonyl groups which are opposite to each other on the same six-carbon ring, and the paradibenzoic dihydrazide has two hydrazide groups which are opposite to each other on the same benzene ring. If the sample contains anthraquinone compounds with a certain concentration, under the catalysis of sulfuric acid, the amino groups on the two opposite hydrazide groups and the two opposite carbonyl groups are complexed to form purple precipitates, and the more the generated purple precipitates are, the darker the color of the precipitates is. If violet precipitate is not generated, the sample may not contain the anthraquinone compound, may contain the anthraquinone compound at a low concentration, and may contain too little amount of the dibenzoic dihydrazide. Therefore, the excessive paradibenzoic dihydrazide is added to make the anthraquinone compounds react completely, and the method for detecting the anthraquinone compounds can be used for screening out samples containing the anthraquinone compounds with a certain concentration or above. If the anthraquinone compounds contained in the samples reach a certain concentration or more, for example, the mass concentration reaches more than 0.05%, respectively 0.05%, 0.06% and the like, and excessive paradibenzoic acid dihydrazide is not added, for example, after the paradibenzoic acid dihydrazide is added, the mass concentration of the paradibenzoic acid dihydrazide in the reaction solution is 0.01%, only the same amount of anthraquinone compounds can be reacted, and a purple precipitation phenomenon can be observed, but the precipitation color depth is the same, which is not beneficial to distinguishing the concentration difference of the anthraquinone compounds of the samples.
The detection method can be applied to screening fungus and fermentation conditions capable of producing anthraquinone compounds at high yield. For various fungus fermentation broths, the method for detecting anthraquinone compounds is utilized, catalytic amount of sulfuric acid and excessive paradibenzoic dihydrazide are added into the fungus liquid, the darker the color of the generated purple precipitate is, the thicker the anthraquinone compounds are contained, the same volume of fungus liquid sample is taken, the fungus capable of relatively high producing anthraquinone compounds and the fermentation conditions of the fungus can be identified by adopting the detection method, the fungus can be screened, and the fermentation conditions are used for further expanding culture fermentation, so that the productivity is improved.
Detailed Description
The method for detecting anthraquinone compounds and its application are specifically described below.
Some information about the anthraquinone-based chemical and the p-dibenzoic dihydrazide used in the present embodiment are shown in table 1 below.
TABLE 1 structural formulas and colors of anthraquinone-based chemicals and p-dibenzoic dihydrazide
Examples 1 to 21 below were seven anthraquinone compounds of Table 1, sulfuric acid and paradibenzoic acid dihydrazide were added thereto, and the reaction was observed. Examples 1 to 7 were carried out with the difference that the types of anthraquinone compounds were different. The anthraquinone compounds used in examples 7 to 9 were identical, except that the sulfuric acid concentrations used were different. The sulfuric acid and the paradibenzoic dihydrazide charged in each example are in excess relative to the anthraquinone.
Example 1
Anthraquinone solution with concentration of 0.0001g/mL is prepared, and the solvent is dichloromethane.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
1ML of the anthraquinone solution with the configuration of about 1g is taken, 0.01mL of 37% sulfuric acid is added into the anthraquinone solution, 0.03mL of the paradibenzoic dihydrazide solution with the configuration is added into the anthraquinone solution, the mixture is oscillated for 1min, purple precipitation is gradually generated within 1min, and the phenomenon of purple precipitation is observed to reach a more obvious degree by naked eyes within about 1 min.
Example 2
1-Hydroxyanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1-hydroxyanthraquinone solution prepared in the above way, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the above way, oscillating for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate phenomenon to reach a more obvious degree about 1min by naked eyes.
Example 3
1-Chloroanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1-chloroanthraquinone solution prepared in the above way, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the above way, oscillating for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate phenomenon to reach a more obvious degree about 1min by naked eyes.
Example 4
1-Aminoanthraquinone solution at a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1-aminoanthraquinone solution prepared in the above way, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the above way, oscillating for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate phenomenon to reach a more obvious degree about 1min by naked eyes.
Example 5
1, 5-Diaminoanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1, 5-diaminoanthraquinone solution prepared in the above way, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic acid dihydrazide solution prepared in the above way, oscillating for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate phenomenon to reach a more obvious degree about 1min by naked eyes.
Example 6
1,3, 8-Trihydroxy-6-methylanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1,3, 8-trihydroxy-6-methylanthraquinone solution prepared in the above way, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the above way, oscillating for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate phenomenon to reach a more obvious degree about 1min by naked eyes.
Example 7
1, 5-Dihydroxy-3-methoxy-7-methylanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1, 5-dihydroxyl-3-methoxy-7-methylanthraquinone solution, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution, shaking for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate to reach a remarkable degree about 1min by naked eyes.
Example 8
1, 5-Dihydroxy-3-methoxy-7-methylanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1, 5-dihydroxyl-3-methoxy-7-methylanthraquinone solution, adding 0.01mL of 10% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution, shaking for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate to reach a remarkable degree about 1min by naked eyes.
Example 9
1, 5-Dihydroxy-3-methoxy-7-methylanthraquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the 1, 5-dihydroxyl-3-methoxy-7-methylanthraquinone solution, adding 0.01mL of 70% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution, shaking for 1min, gradually generating purple precipitate within 1min, and observing the purple precipitate to reach a remarkable degree about 1min by naked eyes.
The reaction phenomena in examples 7 to 9 were not significantly different, and it was found that the catalytic effect on the above precipitation reaction was substantially the same by using sulfuric acid having a mass concentration of 10 to 70%, and the reaction phenomena were not significantly different.
Examples 10 to 12 below were conducted to detect anthraquinone compounds in a mixture of plural anthraquinone compounds.
Example 10
A mixed solution containing anthraquinone with a concentration of 0.0001g/mL and 1-hydroxy anthraquinone with a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the mixed solution prepared in the embodiment, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the embodiment, oscillating for 1min, gradually generating deep purple precipitate within 1min, and observing the phenomenon of deep purple precipitate to reach a more obvious degree about 1min by naked eyes.
Example 11
A mixed solution containing 1-chloroanthraquinone at a concentration of 0.0001g/mL, 1-aminoanthraquinone at a concentration of 0.0001g/mL, and 1, 5-diaminoanthraquinone at a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the mixed solution prepared in the embodiment, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the embodiment, oscillating for 1min, gradually generating deep purple precipitate within 1min, and observing the phenomenon of deep purple precipitate to reach a more obvious degree about 1min by naked eyes.
Example 12
A mixed solution containing 1,3, 8-trihydroxy-6-methylanthraquinone at a concentration of 0.0001g/mL and 1, 5-dihydroxy-3-methoxy-7-methylanthraquinone at a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Taking 1mL of the mixed solution prepared in the embodiment, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of the paradibenzoic dihydrazide solution prepared in the embodiment, oscillating for 1min, gradually generating deep purple precipitate within 1min, and observing the phenomenon of deep purple precipitate to reach a more obvious degree about 1min by naked eyes.
As is clear from examples 1 to 12, the purple precipitation phenomenon can be generated by adding the anthraquinone compound with sulfuric acid and paradibenzoic acid dihydrazide, and the anthraquinone compound with a concentration of 0.0001g/mL or more can be detected by adding the sulfuric acid and paradibenzoic acid dihydrazide in a proper concentration and addition amount. Wherein examples 10 and 12 produced a deep purple precipitate having a color depth greater than the purple precipitate produced in examples 1-9. Example 11 produced a deep purple precipitate with a color depth greater than the purple precipitate produced in examples 10 and 12. It can be seen that the higher the concentration of anthraquinone compounds, the darker the color of the purple precipitate formed, in the case of excess sulfuric acid and p-dibenzoic dihydrazide. Thus, the conclusion can be used to compare the concentration of anthraquinone compounds in different samples.
Examples 13 to 19
Examples 13 to 19, which correspond to examples 1 to 7, were set to reduce the concentrations of the anthraquinone compounds, respectively, from 0.0001g/mL to 0.00001g/mL, and the other conditions were the same, and no purple precipitation was observed.
All of the above examples 1-19 were conducted at room temperature of 20-25 ℃ with the addition of the water bath heating means of example 20 below.
Example 20
In this example, the water bath heating conditions were increased as compared with example 1, specifically as follows.
Anthraquinone solution with concentration of 0.0001g/mL is prepared, and the solvent is dichloromethane.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
Under the water bath environment of 50 ℃, 1mL of anthraquinone solution with the configuration of about 1g is added into a test tube, 0.01mL of 37% sulfuric acid is added, 0.03mL of paradibenzoic acid dihydrazide solution with the configuration is added, purple precipitation is gradually generated within 5s, and the phenomenon of purple precipitation is observed to be more obvious about 5s by naked eyes.
From the above example 20, it can be seen that heating can accelerate the precipitation reaction, and improve the detection efficiency.
Table 2 below shows the names and structural formulas of the chemicals used in comparative examples 1 to 4 below.
Table 2 sample structures and colors for the test of comparative examples 1-4
Comparative example 1
A p-benzoquinone solution with a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
1ML of the p-benzoquinone solution prepared in the above way is taken, 0.01mL of 37% sulfuric acid is added, 0.03mL of the p-dibenzoic dihydrazide solution prepared in the above way is added, and the solution is oscillated for 1min, so that no precipitate is generated, and no obvious reaction phenomenon exists.
Comparative example 2
1, 4-Naphthoquinone solution with concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
1ML of the 1, 4-naphthoquinone solution prepared in the above way is taken, 0.01mL of 37% sulfuric acid is added, 0.03mL of the paradibenzoic acid dihydrazide solution prepared in the above way is added, and the shaking is carried out for 1min, so that no precipitate is generated, and no obvious reaction phenomenon exists.
Comparative example 3
A9, 10-phenanthrenequinone solution with a concentration of 0.0001g/mL is prepared, and the solvent is dichloromethane.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
1ML of the 9, 10-phenanthrenequinone solution prepared in the above way is taken, 0.01mL of 37% sulfuric acid is added, 0.03mL of the paradibenzoic acid dihydrazide solution prepared in the above way is added, and the shaking is carried out for 1min, so that no precipitate is generated, and no obvious reaction phenomenon exists.
Comparative example 4
An anthrone solution with a concentration of 0.0001g/mL was prepared, and the solvent was methylene chloride.
A solution of paradibenzoic acid dihydrazide with a concentration of 0.04g/mL is prepared, and the solvent is ethanol.
1ML of the anthrone solution prepared above is taken, 0.01mL of 37% sulfuric acid is added, 0.03mL of the paradibenzoic dihydrazide solution prepared above is added, and the solution is oscillated for 1min, so that no precipitate is generated, and no obvious reaction phenomenon exists.
By combining the above examples 1 to 20 and comparative examples 1 to 4, the purple precipitation phenomenon occurs by adding excessive sulfuric acid and excessive paradibenzoic acid dihydrazide to various anthraquinone compounds with concentration of 0.0001g/mL or more, while the purple precipitation phenomenon does not occur by adding sulfuric acid and paradibenzoic acid dihydrazide to similar anthraquinone compounds such as benzoquinone, 1, 4-naphthoquinone, 9, 10-phenanthrenequinone, anthrone and the like, and the sulfuric acid and paradibenzoic acid dihydrazide have high selective reactivity to anthraquinone compounds. Thus, by adding sulfuric acid and paradibenzoic dihydrazide to the sample, if purple precipitation occurs, the sample can be primarily judged to contain anthraquinone compounds. In order to improve the accuracy of the determination result and to detect the type and concentration of the anthraquinone compounds, a gas chromatography-mass spectrometer (GC-MS) is also used, namely, a gas chromatography tandem mass spectrometry is used for retesting, the detection method refers to a detection method of anthraquinone residue in SN/T4777-2017 outlet tea leaves, and instrument parameters refer to relevant parameters measured by SVOCs. The gas chromatography tandem mass spectrometry detection results were reliable, and the accuracy of the preliminary determination could be further verified, as in examples 21 to 22 and application example 1 below.
Example 21
Weighing 1g of dry rheum officinale traditional Chinese medicine, grinding into powder, heating, refluxing and leaching by using 100mL of dichloromethane, performing nitrogen blowing concentration on leaching solution to 10mL by using a nitrogen blowing instrument, taking 1mL of concentrated solution, adding 0.01mL of 37% sulfuric acid, adding 0.03mL of paradibenzoic acid dihydrazide solution (the solvent is ethanol) with the concentration of 0.04g/mL, oscillating for 1min, gradually generating deep purple precipitate within 1min, and observing the phenomenon of deep purple precipitate to reach a more obvious degree about 1min by naked eyes, wherein the color depth is equivalent to that of the deep purple precipitate generated in example 11.
The residual concentrated solution was retested using a gas chromatograph-mass spectrometer (GC-MS) and was detected to contain anthraquinone compounds at the following concentrations: 0.0012g/mL of chrysophanol, 0.0009g/mL of emodin, 0.0007g/mL of physcion, 0.0004g/mL of aloe-emodin and 0.0003g/mL of rhein. The total concentration of anthraquinone compounds in the concentrated solution is 0.0035g/mL.10mL of the concentrate contained 0.035g of the anthraquinone compound in total. The dried radix et rhizoma Rhei Chinese medicinal material contains 3.5% anthraquinone compounds.
Example 22
Squeezing fresh Chinese cabbage, filtering, collecting 1mL filtrate, adding 37% sulfuric acid 0.01mL, adding 0.03mL of 0.04g/mL paradibenzoic acid dihydrazide solution (ethanol as solvent), shaking for 1min, and no precipitation is observed. The remaining concentrate was retested using a gas chromatograph-mass spectrometer (GC-MS) and no anthraquinone compound was detected.
The detection method for determining that the sample contains anthraquinone compounds is applied to fungus tests for screening high-yield anthraquinone compounds.
Application example 1
The method for detecting the anthraquinone compounds, which is summarized in the above examples and comparative examples, is applied to detection of the anthraquinone compounds in various fungus fermentation broths, and fungus and fermentation conditions corresponding to samples which generate purple precipitates and have dark precipitation colors are screened out, so that the fungus and fermentation conditions capable of producing the anthraquinone compounds at high yield are obtained.
In the same strain, the fermentation conditions are the same with pH and nutrients as variables, and other fermentation conditions are the same, and the most suitable fermentation conditions are selected according to the strain. The fermentation time of fusarium oxysporum, tabasheer bacteria, penicillium and bacillus subtilis is 10 days, and the temperature of bacterial liquid is kept at 26-30 ℃. The fermentation time of staphylococcus aureus and escherichia coli is 10 days, and the temperature of bacterial liquid is kept at 36-38 ℃.
Before detection, each fungus fermentation liquid is crushed by a high-pressure homogenizer to release intracellular liquid. And separating cell fragments from the liquid by adopting a centrifugal machine, and obtaining supernatant serving as a sample to be detected.
The reagents used were: 0.04g/mL of paradibenzoic dihydrazide solution, and ethanol as a solvent; 37% sulfuric acid.
In the detection process, 1mL of supernatant of each fungus fermentation broth, 0.01mL of sulfuric acid and 0.03mL of paradibenzoic acid dihydrazide solution are taken.
The detection results are shown in Table 3 below.
TABLE 3 detection table of anthraquinone compounds produced by various strains and fermentation conditions thereof
/>
/>
As can be seen from the results of the examination in Table 3, the anthraquinone compound in the solution having a mass concentration of 0.01% or more, but the anthraquinone compound in the solution having a mass concentration of 0.001% or more could not be detected by adding a catalytic amount of sulfuric acid and an excessive amount of paradibenzoic dihydrazide solution to the solution containing the anthraquinone compound. After adding sulfuric acid and a solution of paradibenzoic dihydrazide to the sample, if a violet precipitate is formed, the sample contains anthraquinone compounds. If no violet precipitate is produced, the sample may not contain the anthraquinone compound, but may also contain a low concentration of the anthraquinone compound. The method for detecting the anthraquinone compounds can be used for screening bacterial liquid samples containing the anthraquinone compounds with a certain concentration, so that bacterial strains with high yield of the anthraquinone compounds and fermentation conditions are screened out: the penicillin in Table 3, the fermentation conditions are pH 6.5-7.0, and the nutrient used for fermentation is agar; the tabasheer bacteria shown in Table 3 have pH of 5.0-5.5, and the nutrient used for fermentation is corn paste.
In the practical application process, the sample without purple precipitation phenomenon is not needed to be retested by gas chromatography tandem mass spectrometry.
In the embodiment, sulfuric acid and paradibenzoic acid dihydrazide are added into a sample, and a method for observing whether purple precipitation is generated is used for primarily judging whether the sample contains anthraquinone compounds or not, and the detection method is applied to screening fungus and fermentation conditions capable of producing the anthraquinone compounds in high yield, and particularly for various fungus fermentation broths, the detection efficiency can be remarkably improved by using the method for detecting the anthraquinone compounds. The method is characterized in that sulfuric acid with catalytic amount and excessive paradibenzoic dihydrazide are added into bacterial liquid, the deeper the color of the generated purple precipitate is, the thicker the anthraquinone compounds are contained, the bacterial liquid sample with the same volume is taken, the bacteria capable of producing the anthraquinone compounds relatively high and the fermentation conditions of the bacteria can be identified by adopting the detection method, the bacteria can be screened, and the fermentation conditions are used for further culture and fermentation, so that the required anthraquinone compounds can be produced efficiently.
The above is merely a preferred embodiment of the present application, the protective scope of the present application is not limited to the above examples, it should be noted that modifications and alterations will be apparent to those skilled in the art without departing from the principle of the present application, and these modifications and alterations should also be regarded as falling within the protective scope of the present application.
Claims (8)
1. A method for detecting anthraquinone compounds, said method comprising:
The anthraquinone compound has two carbonyl groups which are relatively aligned on the same six-carbon ring, and the carbon of two carbonyl groups is derived from the six-carbon ring;
Sulfuric acid and paradibenzoic dihydrazide are added into a plurality of samples to be detected, and if purple precipitation is generated, the samples are primarily judged to contain the anthraquinone compounds.
2. The method for detecting an anthraquinone compound according to claim 1, wherein after preliminary determination of the anthraquinone compound contained in the sample, the type and concentration of the anthraquinone compound in the sample are also tested using gas chromatography tandem mass spectrometry.
3. The method for detecting anthraquinone compounds according to claim 1 or 2, characterized in that the sulfuric acid and the paradibenzoic acid dihydrazide are added into a sample to be detected at a temperature of 50-60 ℃ during detection.
4. The method for detecting anthraquinone compounds according to claim 1, wherein the anthraquinone compounds are one or more of anthraquinone, 1-hydroxyanthraquinone, 1-chloroanthraquinone, 1-aminoanthraquinone, 1, 5-diaminoanthraquinone, 1,3, 8-trihydroxy-6-methylanthraquinone and 1, 5-dihydroxy-3-methoxy-7-methylanthraquinone.
5. The method for detecting anthraquinone compounds according to claim 1, characterized in that said sample is, by mass: the sulfuric acid: the p-dibenzoic dihydrazide=1: (0.01 to 0.2): (0.001 to 0.03).
6. Use of the method for detecting anthraquinone compounds according to any of claims 1-5, characterized in that the method for detecting anthraquinone compounds is applied to screening fungus and fermentation conditions of relatively high yield anthraquinone compounds; the sample is taken from fungus fermentation liquor; and adding a catalytic amount of sulfuric acid and an excessive amount of paradibenzoic dihydrazide into the sample, and screening out fungus corresponding to one or more samples with the highest precipitation color and fermentation conditions of the fungus.
7. The method for detecting anthraquinone compounds according to claim 6, wherein before adding sulfuric acid and paradibenzoic dihydrazide, fungus fermentation liquid is subjected to fungus crushing and centrifugation to obtain supernatant, the obtained supernatant is directly used as the sample to be detected, or the obtained supernatant is extracted and then an extract is used as the sample to be detected.
8. The application of the method for detecting anthraquinone compounds according to claim 7, characterized in that the supernatant is directly used as the sample to be detected in a titration mode, and the paradibenzoic acid dihydrazide added into the sample to be detected is added in the form of paradibenzoic acid dihydrazide solution with the concentration of 0.04-0.1 g/mL; the solvent of the paradibenzoic acid dihydrazide solution is ethanol; by volume, the sample: the paradibenzoic dihydrazide solution = 1: (0.03 to 0.20).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311335576.9A CN117420256B (en) | 2023-10-16 | 2023-10-16 | Method for detecting anthraquinone compounds and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311335576.9A CN117420256B (en) | 2023-10-16 | 2023-10-16 | Method for detecting anthraquinone compounds and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117420256A CN117420256A (en) | 2024-01-19 |
| CN117420256B true CN117420256B (en) | 2024-06-04 |
Family
ID=89529338
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311335576.9A Active CN117420256B (en) | 2023-10-16 | 2023-10-16 | Method for detecting anthraquinone compounds and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117420256B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105424855A (en) * | 2015-11-23 | 2016-03-23 | 浙江出入境检验检疫局检验检疫技术中心 | Method for measuring residual amount of anthraquinone in tea with gas chromatography-mass spectrum/mass spectrum method |
| CN108680669A (en) * | 2018-05-08 | 2018-10-19 | 江苏安舜技术服务有限公司 | The test method of anthraquinone in a kind of plant-derived product and food |
| CN109781491A (en) * | 2019-03-18 | 2019-05-21 | 上海国齐检测技术有限公司 | A kind of pre-treating method that PAHs in soil extracts |
| CN113514591A (en) * | 2021-09-02 | 2021-10-19 | 江南大学 | Method for extracting anthraquinone compounds |
-
2023
- 2023-10-16 CN CN202311335576.9A patent/CN117420256B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105424855A (en) * | 2015-11-23 | 2016-03-23 | 浙江出入境检验检疫局检验检疫技术中心 | Method for measuring residual amount of anthraquinone in tea with gas chromatography-mass spectrum/mass spectrum method |
| CN108680669A (en) * | 2018-05-08 | 2018-10-19 | 江苏安舜技术服务有限公司 | The test method of anthraquinone in a kind of plant-derived product and food |
| CN109781491A (en) * | 2019-03-18 | 2019-05-21 | 上海国齐检测技术有限公司 | A kind of pre-treating method that PAHs in soil extracts |
| CN113514591A (en) * | 2021-09-02 | 2021-10-19 | 江南大学 | Method for extracting anthraquinone compounds |
Non-Patent Citations (3)
| Title |
|---|
| 张志贤 等.《有机官能团定量分析》.化学工业出版社,1990,第206页. * |
| 氯代和溴代二苯并环庚烯类高选择性杀鼠剂的合成及生物活性研究;刘丽萍;《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》;20201202;第24页 * |
| 王薇 等.《中药化学实验指导》.陕西科学技术出版社,2014,第80页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117420256A (en) | 2024-01-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Monteiro et al. | Ethyl carbamate formation in wine: Use of radioactively labeled precursors to demonstrate the involvement of urea | |
| Rico-Yuste et al. | Analysis of alternariol and alternariol monomethyl ether in foodstuffs by molecularly imprinted solid-phase extraction and ultra-high-performance liquid chromatography tandem mass spectrometry | |
| CN109358043B (en) | Method for rapidly detecting pesticide residues by using organic solvent extraction sample | |
| CN113624752B (en) | Aptamer-based colorimetric sensor for enhancing activity of MOFs enzymes and preparation method and application thereof | |
| Ghiaba et al. | Study of antioxidant properties of five Algerian date (phoenix dactylifera L) cultivars by cyclic voltammetric technique | |
| CN117420256B (en) | Method for detecting anthraquinone compounds and application thereof | |
| Chen et al. | A simple “turn-on” fluorescence sensor for salicylaldehyde skeleton based on switch of PET-AIE effect | |
| CN108267437B (en) | Based on unicellular Raman spectrum sum15N2Method for detecting stable isotope labeled nitrogen-fixing bacteria | |
| Cao et al. | Rapid quantification of intracellular polyhydroxyalkanoates via fluorescence techniques: A critical review | |
| CN106086211A (en) | LAMP primer group, test kit and the detection method of detection Flos Nelumbinis corruption pathogenic bacteria | |
| CN116660425B (en) | Method for rapidly determining various pesticide residues in fruits and vegetables and application thereof | |
| CN109030440B (en) | Method for detecting content of tannic acid based on molybdenum trioxide quantum dots | |
| CN108707697A (en) | The LAMP detection primer pair and detection method of Sai Nika paddy viruses | |
| Duan et al. | A colorimetric and ratiometric fluorescent probe for rapid, sensitive and visual detection of metabisulfite in food and living cells | |
| CN108315471A (en) | Identify specific gene, specific primer, the kit containing the primer and its application of clubroot germina number-four biological strain | |
| CN107513562A (en) | Detect pseudomonas aeruginosa and ExoU primer, kit and method in water | |
| CN106520914A (en) | Primers and probe, kit and method used for precise and quantitative detection of ovine-derived materials | |
| CN109868329A (en) | Colletotrichum specific primer screening quarantine identification method | |
| Cheng et al. | Molecular identification of wines using in situ liquid SIMS and PCA analysis | |
| CN112321489B (en) | Electrophilic molecular probe based on active sulfydryl and report ions and preparation method and application thereof | |
| CN110596083B (en) | Reagent and card for detecting organophosphorus and carbamate pesticide residues | |
| CN108467897A (en) | Detect the primer of C.perfringens and plc genes, kit and method in food | |
| CN108279223B (en) | Fluorescence detection method for detecting sinapine thiocyanate based on cationic polymer | |
| Krog | On Bryoria chalybeiformis and some related species | |
| CN106525806A (en) | Method for detecting ethrel by using fluorescent nitrogen doped carbon quantum dot |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |