CN105203687A - Paeonol quick detection method - Google Patents
Paeonol quick detection method Download PDFInfo
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- CN105203687A CN105203687A CN201510598976.8A CN201510598976A CN105203687A CN 105203687 A CN105203687 A CN 105203687A CN 201510598976 A CN201510598976 A CN 201510598976A CN 105203687 A CN105203687 A CN 105203687A
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Abstract
The invention provides a paeonol quick detection method. The method includes the steps that firstly, a sample is prepared; secondly, PY-GC/MS detection is carried out; thirdly, data analysis and qualitative and quantitative analysis are carried out. The content of paeonol is detected through the PY-GC/MS technology, the paeonol quick and trace detection method is provided according to moutan bark, cynanchum paniculatum and other traditional Chinese medicine with paeonol as the main active component in actual production and life to solve the problem that quality is different due to different production places, seasons and preparation, and accordingly the quality of traditional Chinese medicine of the kind is primarily judged. The method has the advantages of being easy to operate, low in detection limit, good in experimental reproducibility, visual and reliable in experiment data and the like, is particularly suitable for trace sample detection, can be used for primary comparison of quality differences between the moutan bark, the cynanchum paniculatum and other samples in different production places, varieties and batches, and is wide in applicability.
Description
Technical field
The present invention relates to a kind of method detecting paeonol content based on fast pyrogenation-Gas chromatographyMass spectrometry (PY-GC/MS method) fast.
Background technology
Paeonol (Paeonol), also known as paeonol, is the principle active component of the dry root of ranunculaceae peony root skin and trailing plants Mi section plant paniculate swallowwort or herb.There is calmness, hypnosis, antipyretic, analgesia, anti-inflammatory, the pharmacological action such as antibacterial.
Nearest research shows that Paeonol has good preventive and therapeutic action in cardiovascular and cerebrovascular disease.Paeonol and aspirin compare the effect changing rat blood rheological by Li Wei etc., show: Paeonol can reduce whole blood apparent viscosity, packed cell volume, erythrocyte aggregation and platelet adhesion reaction, erythrocytic deformability is significantly strengthened; The ability that aspirin changes hemorheological property is then comparatively limited to.Illustrate that Paeonol has intervention effect at thrombotic links thus.In addition, Paeonol can the formation of dose-dependent suppression strong carcinogen, and especially in the process that aromatic amine compounds is carcinogenic, serve the effect of Tumor suppression growth, Paeonol and derivant thereof are expected to develop the antineoplastic becoming a new generation.Therefore, the Paeonol fast, in efficient detection Chinese medicine or its compound preparation is very important to druggability and modernization of Chinese medicine application.
The existing analysis to Paeonol detects the main method adopting efficient liquid phase.By the restriction of analytical instrument, often need to carry out comparatively complicated pre-treatment to sample, by methods such as extractions, Paeonol is separated from medicinal material or its compound preparation, adopts the liquid-phase chromatography method analyses such as HPLC/UPLC/SFCLC subsequently.The sample handling processes of this analytical approach is complicated, the aggregate analysis time is long.
Fast pyrogenation-gas chromatography/mass spectrometry technology (PY-GC/MS) is a kind of technology of efficient detection effumability composition, and development in recent years is rapid.Pyrolytic process under low temperature is a kind of very effective thermal release means, can be realized preliminary solution-air or the vapor solid separation of lower boiling complex mixture by the temperature and time controlling pyrolysis; For gas chromatography provide a kind of online with real-time sample pre-treatments and initial gross separation ability.Pyrolysis time in analyte detection process, temperature, heating rate, flow rate of carrier gas, split ratio, ionization temperature etc. are very large on the impact of analysis result.Therefore, based on the feature of PY-GC/MS technology, need badly at present and propose a kind of quick, Sparklet testing method for Paeonol.
Summary of the invention
The object of the present invention is to provide a kind of method for quick of Paeonol, and based on this, tentatively judge the height of the Chinese crude drug such as moutan bark, paniculate swallowwort and the quality of the pharmaceutical preparations thereof.
For achieving the above object, present invention employs following technical scheme:
1) accurately take moutan bark, paniculate swallowwort or its compound preparation 0.02 ~ 5.00mg and obtain sample, sample is added in pyrolysis cup;
2) fast pyrogenation-gas chromatograph-mass spectrometer (GCMS) is adopted to measure the sample added in pyrolysis cup;
3) through step 2) after, according to the original chromatographic data measured and mass spectrometric data, the chromatographic peak of Paeonol in qualitative sample, and obtain the relative content of Paeonol in sample.
Described sample is block or pulverulent solids.
The condition of described pyrolysis is: pyrolysis temperature is 180 ~ 300 DEG C, and pyrolysis time is 0.1 ~ 2min.
The condition of work of described gas chromatography is: carrier gas is the helium of flow velocity 0.1 ~ 4.0mL/min, and split ratio is 0 ~ 500:1, and heating schedule is: initial temperature is 40 ~ 70 DEG C, and keeps 0 ~ 3min, then rises to 300 DEG C with≤20 DEG C/min.
Described mass spectrographic condition of work is: adopt EI source, positive ion detection, electron energy is 50 ~ 85eV, and ion source temperature is 215 ~ 245 DEG C, single level Four bar temperature is 145 ~ 185 DEG C, scan mode is full scan mode, and electron-multiplier voltage is 1075 ~ 2980V, and solvent delay is 0 ~ 6min.
Described relative content adopts area normalization method to calculate.
The qualitative employing NIST08 software Auto-matching of the chromatographic peak of described Paeonol calculates.
Beneficial effect of the present invention is embodied in:
1) quick separating.Existing method is that moutan bark or paniculate swallowwort or its preparation are obtained Paeonol solution by extracting process initial gross separation, after filtration treatment, utilizes liquid-phase chromatographic analysis, or thin-layer chromatography-spectrum coupling technique (TLC-SP) etc. detects its effective constituent; The general analysis time of the method is 4 ~ 24 hours.Directly initial gross separation is carried out to the low-boiling point material of moutan bark or paniculate swallowwort or its preparation by thermal release method in the present invention, gas chromatography is adopted to be separated Paeonol further subsequently, greatly save analysis time, the conventional analysis time is 0.1 ~ 0.3 hour, improves analysis efficiency.
2) sensitivity is good, and required sample size is few.HPLC detection method is subject to the restriction of instrument, generally needs first to be separated to obtain crude product solution, and the sample size therefore needed is comparatively large, and normally several grams to tens grams not etc.; The analysis specimen in use amount of the inventive method is generally 0.02 ~ 1mg, improves sensitivity for analysis.
3) present invention optimizes the testing conditions such as pyrolysis, chromatogram, mass spectrum, can chromatographic peak needed for fast qualitative, achieve and detect fast and accurately.
Embodiment
Below in conjunction with embodiment, the invention will be further described.
The present invention is directed to Paeonol and melt the relatively low and volatile feature of boiling point, adopt fast pyrogenation-gas chromatography/mass spectrometry technology (PY-GC/MS), fully thermal release and chromatographic resolution are combined, realize the quick separating of Paeonol, adopt mass spectrometry subsequently and quantitatively detect, reaching object that is quick, trace detection.
Embodiment
Detect a method for Paeonol based on PY-GC/MS, for Chinese crude drug moutan bark, but protection scope of the present invention is not limited to this embodiment, and concrete steps are as follows:
One, preparation of samples
The moutan bark obtained (Shaanxi Yu Wangtang biotechnology Development Co., Ltd, Xi'an) is accurately taken the pyrolysis cuvette that 1.00mg (± 0.02mg) puts into 50 μ L;
Two, pyrolysis:
Pyrocrack furnace adopts the PY2020is of Frontiers;
1. pyrolysis temperature: 200 DEG C;
2. pyrolysis time: 0.20min.
Three, be separated, detect
Chromatogram:
Gas chromatography adopts Agilent 7890A:
1. analytical column is HP-5MS fused-silica capillary column (30m*0.25id*0.25 μm);
2. carrier gas: high-purity helium, flow velocity is 1.0mL/min;
3. input mode is split sampling, and split ratio is 150:1;
4. injector temperature is 280 DEG C, and transmission line temperature is 290 DEG C;
5. temperature programme: initial temperature is 60 DEG C, keeps 1min, then rises to 300 DEG C with 20 DEG C/min.
Mass spectrum (adopting Agilent 5975C):
1. EI source, positive ion detects, and electron energy is 70eV;
2. ion source temperature is 230 DEG C, and single level Four bar temperature is 150 DEG C;
3. scan mode is full scan mode;
4. electron-multiplier voltage is 2042V, and solvent delay is 3min.
Four, data analysis and process
Retention time and the integral area of chromatogram are completed automatically by workstation, utilize NIST08 to compose the qualitative chromatographic peak in storehouse.The relative content of each volatile ingredient is obtained by area normalization method.
In this sample moutan bark pyrolyzed components, identifiable peak is 23, and composition and retention time thereof list in table 1.Retention time and peak area Agilent Chemstation calculate, and the method for relative content area normalization calculates.
The composition of volatile ingredient and content (t in table 1. moutan bark
r=6.999: Paeonol)
Five, precision, reappearance and stability experiment
1. Precision Experiment: to take from 5 parts, the sample of same moutan bark, continuous sample introduction 5 times according to the method described above, 23 chromatographic peaks relative peak area being greater than to 0.5% are analyzed, the RSD (relative standard deviation) of its relative retention time and relative peak area, respectively between 1.25 ~ 2.35% and 1.53 ~ 4.33%, illustrates that the inventive method precision is good.
2. reappearance experiment: 5 parts, the sample getting same batch of moutan bark, according to the method described above continuous sample introduction 5 times.The relative retention time of 23 chromatographic peaks and the RSD of relative peak area are respectively between 0.56 ~ 4.37% and 1.48 ~ 3.99% as a result, illustrate that the inventive method reappearance is good.
3. stability experiment: get 1 part, the sample prepared, respectively 0,2,4,8,12,24h sample introduction measures, the relative retention time of 23 chromatographic peaks and the RSD of relative peak area are all less than 3.36% as a result, and it is stable for illustrating that the inventive method measures in 24h.
Chinese crude drug is because of the difference of the place of production, season, process of preparing Chinese medicine mode, and its water cut, density etc. have trickle difference, and the content then in its compound preparation is also different.Therefore, pyrolysis, the separation of needs are all not identical with analysis condition, draw the condition and range in the present invention accordingly.
The present invention has simple to operate, detectability is low, experiment favorable reproducibility, the features such as experimental data is intuitive and reliable, be specially adapted to micro-example and detect (sample size can be low to moderate 20 μ g), and can be used for carrying out quantitative comparison to containing the difference in quality between Paeonol Chinese crude drug or its formulation samples, application is wide, for the quality standard of such Chinese crude drug and modernization application provide the thinking and countermeasure of a reference.
Claims (7)
1. a method for quick for Paeonol, is characterized in that: comprise the following steps:
1) accurately take moutan bark, paniculate swallowwort or its compound preparation 0.02 ~ 5.00mg and obtain sample, sample is added in pyrolysis cup;
2) fast pyrogenation-gas chromatograph-mass spectrometer (GCMS) is adopted to measure the sample added in pyrolysis cup;
3) through step 2) after, according to the original chromatographic data measured and mass spectrometric data, the chromatographic peak of Paeonol in qualitative sample, and obtain the relative content of Paeonol in sample.
2. the method for quick of a kind of Paeonol according to claim 1, is characterized in that: described sample is block or pulverulent solids.
3. the method for quick of a kind of Paeonol according to claim 1, it is characterized in that: the condition of described pyrolysis is: pyrolysis temperature is 180 ~ 300 DEG C, pyrolysis time is 0.1 ~ 2min.
4. the method for quick of a kind of Paeonol according to claim 1, it is characterized in that: the condition of work of described gas chromatography is: carrier gas is the helium of flow velocity 0.1 ~ 4.0mL/min, split ratio is 0 ~ 500:1, heating schedule is: initial temperature is 40 ~ 70 DEG C, and keep 0 ~ 3min, then rise to 300 DEG C with≤20 DEG C/min.
5. the method for quick of a kind of Paeonol according to claim 1, it is characterized in that: described mass spectrographic condition of work is: adopt EI source, positive ion detection, electron energy is 50 ~ 85eV, ion source temperature is 215 ~ 245 DEG C, single level Four bar temperature is 145 ~ 185 DEG C, scan mode is full scan mode, and electron-multiplier voltage is 1075 ~ 2980V, and solvent delay is 0 ~ 6min.
6. the method for quick of a kind of Paeonol according to claim 1, is characterized in that: described relative content adopts area normalization method to calculate.
7. the method for quick of a kind of Paeonol according to claim 1, is characterized in that: the qualitative employing NIST08 software Auto-matching of the chromatographic peak of described Paeonol calculates.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1188442A2 (en) * | 2000-09-13 | 2002-03-20 | Wei Xiao | Cinnamomi and poria composition, method to prepare the same and uses thereof |
| CN1527053A (en) * | 2003-03-07 | 2004-09-08 | 江苏康缘药业股份有限公司 | Quality control method for rehmannia prepn of six ingredients |
| US20060110468A1 (en) * | 2004-11-24 | 2006-05-25 | Liang Liu | Herbal formulations for arthritis |
| CN100409866C (en) * | 2001-09-13 | 2008-08-13 | 江苏康缘药业股份有限公司 | Chinese herbal medicine composition and preparing method |
| CN102253146A (en) * | 2011-07-14 | 2011-11-23 | 黄石卫生材料药业有限公司 | Determination method of eugenol content in stomach-warming and pain-relieving paste |
| CN102335238A (en) * | 2010-07-22 | 2012-02-01 | 江西济民可信集团有限公司 | Detection method of frankincense and elecampane lithiasis-removing granular traditional Chinese preparation |
| CN102735785A (en) * | 2011-03-30 | 2012-10-17 | 江苏康缘药业股份有限公司 | Method for detecting traditional Chinese medicine composition by using gas chromatography and fingerprint chromatography |
-
2015
- 2015-09-18 CN CN201510598976.8A patent/CN105203687A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1188442A2 (en) * | 2000-09-13 | 2002-03-20 | Wei Xiao | Cinnamomi and poria composition, method to prepare the same and uses thereof |
| CN100409866C (en) * | 2001-09-13 | 2008-08-13 | 江苏康缘药业股份有限公司 | Chinese herbal medicine composition and preparing method |
| CN1527053A (en) * | 2003-03-07 | 2004-09-08 | 江苏康缘药业股份有限公司 | Quality control method for rehmannia prepn of six ingredients |
| US20060110468A1 (en) * | 2004-11-24 | 2006-05-25 | Liang Liu | Herbal formulations for arthritis |
| CN102335238A (en) * | 2010-07-22 | 2012-02-01 | 江西济民可信集团有限公司 | Detection method of frankincense and elecampane lithiasis-removing granular traditional Chinese preparation |
| CN102735785A (en) * | 2011-03-30 | 2012-10-17 | 江苏康缘药业股份有限公司 | Method for detecting traditional Chinese medicine composition by using gas chromatography and fingerprint chromatography |
| CN102253146A (en) * | 2011-07-14 | 2011-11-23 | 黄石卫生材料药业有限公司 | Determination method of eugenol content in stomach-warming and pain-relieving paste |
Non-Patent Citations (6)
| Title |
|---|
| DENG CHUNHUI 等: "Development of microwave-assisted extraction followed by headspace single-drop microextraction for fast determination of paeonol in traditional Chinese medicines", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| DONG LING 等: "Fast determination of paeonol in plasma by headspace solid-phase microextraction followed by gas chromatography–mass spectrometry", 《ANALYTICA CHIMICA ACTA》 * |
| 房杏春 等: "直接蒸发—高分辨气相色谱分析牡丹皮和六味地黄丸中丹皮酚的含量", 《中国药科大学学报》 * |
| 曲家妮 等: "丹皮提取物的SPME－GC/MS分析", 《烟草科技》 * |
| 武子敬 等: "牡丹皮挥发油成分分析", 《通化师范学院学报》 * |
| 禹凤英 等: "闪蒸-气相色谱法测定六味地黄口服液中丹皮酚的含量", 《中成药》 * |
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Application publication date: 20151230 |