CN104198620B - A kind of method of liquid chromatography for measuring azophenlyene content - Google Patents
A kind of method of liquid chromatography for measuring azophenlyene content Download PDFInfo
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- CN104198620B CN104198620B CN201410492703.0A CN201410492703A CN104198620B CN 104198620 B CN104198620 B CN 104198620B CN 201410492703 A CN201410492703 A CN 201410492703A CN 104198620 B CN104198620 B CN 104198620B
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Abstract
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; A kind of method of liquid chromatography for measuring azophenlyene content is provided, the method achieves the quantitative detection of azophenlyene, favorable reproducibility, measurement result accurately, be easy to operation, the detection of azophenlyene can be widely used in; In azophenlyene sample, azophenlyene content is according to following formulae discovery: C=-18.06+2.0292 × 10-7S, in formula: the content of azophenlyene in C----azophenlyene sample, and unit is the peak area of mg/mL, S----liquid chromatographic detection; The present invention is mainly used in azophenlyene context of 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 distils, 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, 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 adopted 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 prepared is long for 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 being provided, the method achieving the quantitative detection of azophenlyene, favorable reproducibility, measurement result accurately, be easy to operation, the detection of azophenlyene can be widely used in.
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 is according to following formulae discovery,
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, stir until azophenlyene sample all dissolves with glass bar, 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 azophenlyene storing solution 1mL, 2mL, 3mL, 4mL and 5mL respectively, be placed in 10mL volumetric flask respectively, be diluted to scale with aniline, shake up, obtains 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 obtained 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 the sample introduction concentration of correspondence, 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).
Described to arrange liquid chromatographic detection condition be chromatographic column is C18(particle diameter 5um, internal diameter 4.6mm, length 250mm) or similar performance pillar.
It is described that to arrange liquid chromatographic detection condition be mobile phase A is 15%(V/V) acetonitrile, 10%(V/V) methyl alcohol, 75%(V/V) high purity water.
It is described that to arrange liquid chromatographic detection condition be 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 determined wavelength is 254nm, flow velocity is 1.5mL/min.
The described liquid chromatographic detection condition that arranges is 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 is compared with prior art had is:
The present invention selects aniline, can not only ensure that sample is dissolved completely, and in testing process, can not cause interference to the testing result of azophenlyene, thus makes the testing result of azophenlyene content more accurate; Adopt method of the present invention, achieve the quantitative detection of azophenlyene, favorable reproducibility, measurement result accurately, be easy to operation, the detection of azophenlyene can be widely used in.
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, stir until azophenlyene sample all dissolves with glass bar, 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 10mL volumetric flask respectively, be diluted to scale with aniline, shake up.Obtain series concentration 2.0962,4.1924,6.2886,8.3848, the azophenlyene solution of 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;
Determined wavelength: 254nm;
Flow velocity: 1.5mL/min;
Adopt quantified by external standard method;
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) sample introduction concentration that the peak area that records is corresponding with it carries out linear regression, and draw out standard working curve, show that equation of linear regression is C=-18.06+2.0292 × 10
-7s, related coefficient is 0.9996.
6) azophenlyene content in azophenlyene sample is calculated 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 (3)
1. a method for liquid chromatography for measuring azophenlyene content, is characterized in that: in azophenlyene sample, azophenlyene content is according to following formulae discovery,
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, pour in 50mL beaker, add a small amount of aniline in batches, stir until azophenlyene standard items all dissolve with glass bar, 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 azophenlyene storing solution 1mL, 2mL, 3mL, 4mL and 5mL respectively, be placed in 10mL volumetric flask respectively, be diluted to scale with aniline, shake up, obtains 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 obtained 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 the sample introduction concentration of correspondence, draw out standard working curve, draw equation of linear regression, formulation;
It is described that to arrange liquid chromatographic detection condition be chromatographic column is the C18 of particle diameter 5um, internal diameter 4.6mm, length 250mm;
It is described that to arrange liquid chromatographic detection condition be mobile phase A is 15%(V/V) acetonitrile, 10%(V/V) methyl alcohol, 75%(V/V) high purity water;
It is described that to arrange liquid chromatographic detection condition be Mobile phase B is 60%(V/V) acetonitrile, 40%(V/V) methyl alcohol;
The described liquid chromatographic detection condition that arranges is adopt condition of gradient elution, as following table,
。
2. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 1, is characterized in that: the volume ratio of described azophenlyene sample quality and aniline is (0.002-2.5): (1-100).
3. the method for a kind of liquid chromatography for measuring azophenlyene content according to claim 1, is characterized in that: the described liquid chromatographic detection condition that arranges is that determined wavelength is 254nm, flow velocity is 1.5mL/min.
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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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| 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)
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| High-performance liquid chromatographic analysis of Pseudomonas aeruginosa phenazines;Ruben O. Fernandez et al.;《Journal of Chromatography A》;19970530;第771卷(第1-2期);99-104 * |
| Thermospray Mass Spectrometric Analysis of Phenazines;David Watson et al.;《BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY》;19881231;第17卷;251-255 * |
| 吩嗪的合成及其应用;茅晓晖 等;《化工生产与技术》;20091231;第16卷(第3期);42-47 * |
| 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;曹柳燕 等;《分析测试学报》;20080430;第27卷(第4期);412-414 * |
| 高效液相色谱法测定吩嗪的含量;唐蓉萍 等;《甘肃石油和化工》;20111231(第4期);46-48 * |
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