CN104198620A - Method for measuring phenazine content by liquid chromatography - Google Patents
Method for measuring phenazine content by liquid chromatography Download PDFInfo
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- CN104198620A CN104198620A CN201410492703.0A CN201410492703A CN104198620A CN 104198620 A CN104198620 A CN 104198620A CN 201410492703 A CN201410492703 A CN 201410492703A CN 104198620 A CN104198620 A CN 104198620A
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- azophenlyene
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Abstract
The invention relates to a detection method of a phenazine content, in particular to a method for measuring a phenazine content by liquid chromatography. The method for measuring the phenazine content by the liquid chromatography realizes quantitative detection of phenazine, has good repeatability and accurate measuring results, is easy to operate and can be widely applied to detection of phenazine. The phenazine content in a phenazine sample is calculated according to the following formula: C=-18.06+2.0292*10-7S, wherein C represents the phenazine content in the phenazine sample in mg/mL, and S represents peak area detected by the liquid chromatography. The method is mainly applied to the phenazine detection.
Description
Technical field
The present invention relates to the detection method of azophenlyene content, be specifically related to a kind of method of liquid chromatography for measuring azophenlyene content.
Background technology
Azophenlyene glassy yellow acicular crystal, easily distillation, is insoluble in water, is slightly dissolved in ethanol, ether, is the basic structure skeleton of a large class phenazine dyes, as organic synthesis reagent and dyestuff intermediate.At present, the detection method of azophenlyene mainly contains: Tang Rongping, and Yang Ronghua etc. adopt high performance liquid chromatography to detect azophenlyene content, and this detection method selects Paeonol as internal standard compound, the internal standard method adopting detects, and the defect of the method is that the purity requirement of internal standard compound matter is higher and sample preparation is more complicated.Also someone adopts chloroform, methylene chloride, ethanol equal solvent to dissolve azophenlyene sample, but adopts these solvents, and the solution preparing is long standing time one, and sample is easily separated out again, affects the accuracy of result.Therefore, be necessary to be improved.
Summary of the invention
In order to overcome the deficiencies in the prior art, a kind of method of liquid chromatography for measuring azophenlyene content is provided, the method has realized the quantitative detection of azophenlyene, favorable reproducibility, measurement result accurately, easy operating, can be widely used in the detection of azophenlyene.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is:
A method for liquid chromatography for measuring azophenlyene content, in azophenlyene sample, azophenlyene content calculates according to following formula,
C=-18.06+2.0292×10
-7S
In formula: the content of azophenlyene in C----azophenlyene sample, unit is mg/mL;
The peak area of S----liquid chromatographic detection.
Described formula is according to following steps gained:
The preparation of a, azophenlyene storing solution: accurately take 2.0962g azophenlyene standard items (being accurate to 0.0001g), pour in 50mL beaker, add a small amount of aniline in batches, with glass bar stir until azophenlyene sample all dissolve, moved in 100mL volumetric flask, clean beaker 2-3 time with aniline, cleaning fluid is transferred in volumetric flask, be settled to scale with aniline;
The preparation of b, serial solution concentration: measure respectively azophenlyene storing solution 1 mL, 2 mL, 3 mL, 4 mL and 5mL, be placed in respectively 10mL volumetric flask, be diluted to scale with aniline, shake up, obtain the azophenlyene solution of variable concentrations;
C, use liquid chromatographic detection peak area: liquid chromatographic detection condition is first set, then by the variable concentrations azophenlyene solution obtaining in step b respectively sample introduction 40uL carry out chromatogram detection, record peak area;
The drafting of d, typical curve: carry out linear regression according to the peak area of record and corresponding sample introduction concentration, draw out standard working curve, draw equation of linear regression, formulation.
Described azophenlyene sample quality (g) is (0.002-2.5) with volume (mL) ratio of aniline: (1-100).
The described liquid chromatographic detection condition that arranges is that chromatographic column is C18(particle diameter 5um, internal diameter 4.6mm, length 250mm) or similar performance pillar.
The described liquid chromatographic detection condition that arranges is that mobile phase A is 15%(V/V) acetonitrile, 10%(V/V) methyl alcohol, 75%(V/V) high purity water.
The described liquid chromatographic detection condition that arranges is that Mobile phase B is 60%(V/V) acetonitrile, 40%(V/V) methyl alcohol.
The described liquid chromatographic detection condition that arranges is that quantivative approach adopts external standard method.
The described liquid chromatographic detection condition that arranges is that 254nm, flow velocity are 1.5mL/min for detecting wavelength.
The described liquid chromatographic detection condition that arranges is to adopt condition of gradient elution, as following table,
| Time min | The volume fraction % of A | The volume fraction % of B |
| 0 | 100 | 0 |
| 25 | 25 | 75 |
| 35 | 0 | 100 |
| 37.5 | 0 | 100 |
The beneficial effect that the present invention compared with prior art had is:
The present invention selects aniline, can not only ensure that sample is completely dissolved, and in testing process, can not cause interference to the testing result of azophenlyene, thereby makes the testing result of azophenlyene content more accurate; Adopt method of the present invention, realized the quantitative detection of azophenlyene, favorable reproducibility, measurement result accurately, easy operating, can be widely used in the detection of azophenlyene.
Embodiment
The present invention is further described by the following embodiment.
Embodiment 1
1) preparation of azophenlyene storing solution: accurately take 2.0962g azophenlyene standard items (being accurate to 0.0001g), pour in 50mL beaker, add a small amount of aniline in batches, with glass bar stir until azophenlyene sample all dissolve, moved in 100mL volumetric flask, clean beaker 2-3 time with aniline, cleaning fluid is transferred in volumetric flask, be settled to scale with aniline.
2) preparation of serial solution concentration: accurately measure respectively azophenlyene storing solution 1,2,3,4,5mL, be placed in respectively 10mL volumetric flask, be diluted to scale with aniline, shake up.Obtain the azophenlyene solution of series concentration 2.0962,4.1924,6.2886,8.3848,10.4810mg/mL.
3) liquid chromatographic detection condition is set:
Chromatographic column: C18(particle diameter 5um, internal diameter 4.6mm, length 250mm);
Mobile phase A: 15%(V/V) acetonitrile, 10%(V/V) methyl alcohol, 75%(V/V) high purity water;
Mobile phase B: 60%(V/V) acetonitrile, 40%(V/V) methyl alcohol;
Detect wavelength: 254nm;
Flow velocity: 1.5mL/min;
Employing external standard method is quantitative;
Condition of gradient elution is as following table:
| Time min | The volume fraction % of A | The volume fraction % of B |
| 0 | 100 | 0 |
| 25 | 25 | 75 |
| 35 | 0 | 100 |
| 37.5 | 0 | 100 |
4) detection of sample: get 2) in azophenlyene solution respectively sample introduction 40uL by 3) in chromatographic condition carry out chromatogram detection, record its peak area S and be respectively: 99166217,110279250,119396893,130565016,140633288.
5) drafting of typical curve: according to 4) the peak area sample introduction concentration corresponding with it that records carries out linear regression, draws out standard working curve, show that equation of linear regression is C=-18.06+2.0292 × 10
-7s, related coefficient is 0.9996.
6) calculate azophenlyene content in azophenlyene sample according to equation of linear regression.
C=?-18.06+2.0292×10
-7S
In formula: the content of azophenlyene in C----azophenlyene sample, unit is mg/mL;
The peak area of S----liquid chromatographic detection.
Claims (9)
1. a method for liquid chromatography for measuring azophenlyene content, is characterized in that: in azophenlyene sample, azophenlyene content calculates according to following formula,
C=-18.06+2.0292×10
-7S
In formula: the content of azophenlyene in C----azophenlyene sample, unit is mg/mL;
The peak area of S----liquid chromatographic detection.
2. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 1, is characterized in that: described formula is according to following steps gained:
The preparation of a, azophenlyene storing solution: accurately take 2.0962g azophenlyene standard items (being accurate to 0.0001g), pour in 50mL beaker, add a small amount of aniline in batches, with glass bar stir until azophenlyene sample all dissolve, moved in 100mL volumetric flask, clean beaker 2-3 time with aniline, cleaning fluid is transferred in volumetric flask, be settled to scale with aniline;
The preparation of b, serial solution concentration: measure respectively azophenlyene storing solution 1 mL, 2 mL, 3 mL, 4 mL and 5mL, be placed in respectively 10mL volumetric flask, be diluted to scale with aniline, shake up, obtain the azophenlyene solution of variable concentrations;
C, use liquid chromatographic detection peak area: liquid chromatographic detection condition is first set, then by the variable concentrations azophenlyene solution obtaining in step b respectively sample introduction 40uL carry out chromatogram detection, record peak area;
The drafting of d, typical curve: carry out linear regression according to the peak area of record and corresponding sample introduction concentration, draw out standard working curve, draw equation of linear regression, formulation.
3. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: described azophenlyene sample quality (g) is (0.002-2.5) with volume (mL) ratio of aniline: (1-100).
4. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is that chromatographic column is C18(particle diameter 5um, internal diameter 4.6mm, length 250mm) or similar performance pillar.
5. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is that mobile phase A is 15%(V/V) acetonitrile, 10%(V/V) methyl alcohol, 75%(V/V) high purity water.
6. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is that Mobile phase B is 60%(V/V) acetonitrile, 40%(V/V) methyl alcohol.
7. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is that quantivative approach adopts external standard method.
8. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is that 254nm, flow velocity are 1.5mL/min for detecting wavelength.
9. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 2, is characterized in that: the described liquid chromatographic detection condition that arranges is to adopt condition of gradient elution, as following table,
。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106543011A (en) * | 2015-09-16 | 2017-03-29 | 中国石油化工股份有限公司 | A kind of 4-aminodiphenylamine produces the processing method of waste material |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103992281A (en) * | 2014-05-29 | 2014-08-20 | 山西翔宇化工有限公司 | Method for extracting high-purity phenazine from RT base (4-Aminodiphenylamine) byproduct waste material |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103992281A (en) * | 2014-05-29 | 2014-08-20 | 山西翔宇化工有限公司 | Method for extracting high-purity phenazine from RT base (4-Aminodiphenylamine) byproduct waste material |
Non-Patent Citations (5)
| Title |
|---|
| DAVID WATSON ET AL.: "Thermospray Mass Spectrometric Analysis of Phenazines", 《BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY》 * |
| RUBEN O. FERNANDEZ ET AL.: "High-performance liquid chromatographic analysis of Pseudomonas aeruginosa phenazines", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| 唐蓉萍 等: "高效液相色谱法测定吩嗪的含量", 《甘肃石油和化工》 * |
| 曹柳燕 等: "多菌灵原药中2个酚嗪类杂质含量的高效液相色谱法测定", 《分析测试学报》 * |
| 茅晓晖 等: "吩嗪的合成及其应用", 《化工生产与技术》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106543011A (en) * | 2015-09-16 | 2017-03-29 | 中国石油化工股份有限公司 | A kind of 4-aminodiphenylamine produces the processing method of waste material |
| CN106543011B (en) * | 2015-09-16 | 2018-08-07 | 中国石油化工股份有限公司 | A kind of processing method of 4-aminodiphenylamine production waste material |
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