CN105203663A - Rapid detection method for alkannin - Google Patents
Rapid detection method for alkannin Download PDFInfo
- Publication number
- CN105203663A CN105203663A CN201510600824.7A CN201510600824A CN105203663A CN 105203663 A CN105203663 A CN 105203663A CN 201510600824 A CN201510600824 A CN 201510600824A CN 105203663 A CN105203663 A CN 105203663A
- Authority
- CN
- China
- Prior art keywords
- alkannin
- sample
- quick
- pyrolysis
- temperature
- 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.)
- Pending
Links
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a rapid detection method for alkannin. The method comprises the steps of 1, sample preparation, 2, PY-GC/MS detection and 3, data analysis and qualitative and quantitative analysis. The PY-GC/MS technology is selected for detecting the content of alkannin in lithospermum, the rapid and microcrystalline detection method for alkannin is provided for solving the problems that qualities of the alkannin in actual production and living are different due to different producing areas, seasons and processing methods, and then the alkannin or the quality of the alkannin compound preparation is preliminarily judged. The rapid detection method has the advantages of being easy to implement, low in detection limit, good in experiment reproducibility and visual and reliable in experimental data, is particularly applicable to micro-sample detection, and can be used for preliminary comparison of quality differences of alkannin or alkannin compound preparation samples of different producing areas, varieties, batches and manufacturers, the application range is wide, and the preliminary judgment basis is provided for quality standard and modernization application of alkannin or alkannin compound preparations.
Description
Technical field
The present invention relates to a kind of method detecting alkannin content based on fast pyrogenation-Gas chromatographyMass spectrometry (PY-GC/MS method) fast.
Background technology
Asian puccoon is one of China's traditional Chinese medicine, its root is used as medicine, and has cooling and activating blood, the function of removing toxic substances promoting eruption, main containing alkannin (shikonin), diethylstilbestrol alkannin (acetyIshikonin), etc. multiple tea phenolic compound, there is pharmacological action widely.The anti-inflammatory of Asian puccoon, antitumor action clearly, are among the peoplely usually used in that treatment is scalded, ulcer and various cancer.Wherein, alkannin is the important of such Chinese medicine and main active, and its design feature is a naphthoquinone compound.Pharmacological experiments also demonstrate that alkannin has obvious anti-inflammatory, the effect such as antimycotic and antitumor.There are bibliographical information alkannin and derivant thereof to have obvious cytotoxic activity to kinds of tumor cells, and the apoptosis of the kinds of tumor cells such as human promyelocytic leukemia HL60 cells and people tissue lymph lymphoma U937 cell can be induced.Therefore alkannin or derivatives thereof is one of drug candidate of anticancer.And the alkannin fast, in efficient detection Chinese medicine or its compound preparation is very important to the antitumaous effect research of such Chinese medicine and modernization application.
The existing analysis to alkannin in Asian puccoon detects the method that will adopt liquid chromatography.Often need to carry out comparatively complicated sample pre-treatments to sample, by methods such as extractions, alkannin 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 alkannin.
Summary of the invention
The object of the present invention is to provide a kind of method for quick of alkannin, and based on this, tentatively judge the height of Asian puccoon and compound preparation quality thereof.
For achieving the above object, present invention employs following technical scheme:
1) accurately take Asian puccoon or its compound preparation 0.1 ~ 5.00mg obtains 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 alkannin in qualitative sample, and obtain the relative content of alkannin in sample.
Described sample is block or pulverulent solids.
The condition of described pyrolysis is: pyrolysis temperature is 200 ~ 400 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, split ratio is 0 ~ 500:1, and heating schedule is: initial temperature is 50 ~ 60 DEG C, and keeps 0 ~ 5min, then rise to 150 DEG C with≤30 DEG C/min, then rise to 300 DEG C with≤15 DEG C/min.
Described mass spectrographic condition of work is: adopt EI source, positive ion detection, electron energy is 50 ~ 80eV, and ion source temperature is 220 ~ 240 DEG C, single level Four bar temperature is 120 ~ 180 DEG C, scan mode is full scan mode, and electron-multiplier voltage is 1200 ~ 2900V, and solvent delay is 0 ~ 4min.
Described relative content adopts area normalization method to calculate.
The qualitative employing NIST08 software Auto-matching of the chromatographic peak of described alkannin calculates.
Beneficial effect of the present invention is embodied in:
1) thermal release and chromatographic resolution combine, and realize quick separating.Existing method is that Asian puccoon or its compound preparation are passed through extraction, after isolating thick product, utilizes liquid chromatography or thin-layer chromatography chromatogram etc. to detect its effective constituent; The general analysis time of the method is 6 ~ 12 hours.Directly initial gross separation is carried out to low-boiling point material in Asian puccoon or its compound preparation by thermal release method in the present invention, gas chromatography is adopted to be separated alkannin further subsequently, the conventional analysis time is 0.5 ~ 1 hour, shortens analysis time greatly, improves analysis efficiency.
2) required sample size is few, and sensitivity is good.Existing method obtains crude product solution because needs are first separated, and the sample size therefore needed is comparatively large, and the analysis specimen in use amount of the inventive method is generally 0.1 ~ 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 alkannin 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 alkannin, adopt mass spectrometry subsequently and quantitatively detect, reaching object that is quick, trace detection.
Embodiment
Detect a method for alkannin based on PY-GC/MS, for Chinese crude drug Asian puccoon, but protection scope of the present invention is not limited to this embodiment, and concrete steps are as follows:
One, preparation of samples
The lithospermum euchromum Royle obtained (Shaanxi Yong Yuan Bioisystech 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: 280 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 80:1;
4. injector temperature is 280 DEG C, and transmission line temperature is 290 DEG C;
5. temperature programme: initial temperature is 50 DEG C, keeps 3min, then rises to 300 DEG C with 15 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 2543V, and solvent delay is 3.75min.
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 Asian puccoon pyrolyzed components, identifiable peak is 32, 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. Asian puccoon
r=15.95 alkannins)
Five, precision, reappearance and stability experiment
1. Precision Experiment: to take from 5 parts, the sample of same Asian puccoon, continuous sample introduction 5 times according to the method described above, 32 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 0.35 ~ 1.34% and 1.22 ~ 4.55%, illustrates that the inventive method precision is good.
2. reappearance experiment: 5 parts, the sample getting same batch of Asian puccoon, according to the method described above continuous sample introduction 5 times.The relative retention time of 32 chromatographic peaks and the RSD of relative peak area are respectively between 0.58 ~ 5.06% and 1.44 ~ 4.87% 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 32 chromatographic peaks and the RSD of relative peak area are all less than 4.09% as a result, and it is stable for illustrating that the inventive method measures in 24h.
Basically the main active of Asian puccoon is alkannin, and it can be used as one of main criterion of Asian puccoon or its compound preparation quality.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 selects PY-GC/MS technology to detect alkannin content, is for the needs in actual production, provides a kind of quick, micro-detection method of alkannin, and then tentatively judges the quality of Asian puccoon or its compound preparation.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 100 μ g), and can be used for carrying out quantitative comparison to the difference in quality of different Asian puccoon or its compound preparation sample room, application is wide, for the quality standard of Asian puccoon or its compound preparation and modernization application provide one can the thinking and countermeasure of reference.
Claims (7)
1. a method for quick for alkannin, is characterized in that: comprise the following steps:
1) accurately take Asian puccoon or its compound preparation 0.1 ~ 5.00mg obtains 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 alkannin in qualitative sample, and obtain the relative content of alkannin in sample.
2. the method for quick of a kind of alkannin according to claim 1, is characterized in that: described sample is block or pulverulent solids.
3. the method for quick of a kind of alkannin according to claim 1, it is characterized in that: the condition of described pyrolysis is: pyrolysis temperature is 200 ~ 400 DEG C, pyrolysis time is 0.1 ~ 2min.
4. the method for quick of a kind of alkannin 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 50 ~ 60 DEG C, and keep 0 ~ 5min, then rise to 150 DEG C with≤30 DEG C/min, then rise to 300 DEG C with≤15 DEG C/min.
5. the method for quick of a kind of alkannin 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 ~ 80eV, ion source temperature is 220 ~ 240 DEG C, single level Four bar temperature is 120 ~ 180 DEG C, scan mode is full scan mode, and electron-multiplier voltage is 1200 ~ 2900V, and solvent delay is 0 ~ 4min.
6. the method for quick of a kind of alkannin 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 alkannin according to claim 1, is characterized in that: the qualitative employing NIST08 software Auto-matching of the chromatographic peak of described alkannin calculates.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510600824.7A CN105203663A (en) | 2015-09-18 | 2015-09-18 | Rapid detection method for alkannin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510600824.7A CN105203663A (en) | 2015-09-18 | 2015-09-18 | Rapid detection method for alkannin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105203663A true CN105203663A (en) | 2015-12-30 |
Family
ID=54951454
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510600824.7A Pending CN105203663A (en) | 2015-09-18 | 2015-09-18 | Rapid detection method for alkannin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105203663A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108918737A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | A kind of pre-treating method and efficient liquid-phase chromatography method measuring alkannin in w/o type Asian puccoon skin care milk |
| CN108918738A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | The pre-treating method and efficient liquid-phase chromatography method of alkannin content in a kind of measurement zicao soft plaster |
| CN108918739A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | A kind of pre-treating method and efficient liquid-phase chromatography method measuring alkannin in O/W type Asian puccoon skin care milk |
-
2015
- 2015-09-18 CN CN201510600824.7A patent/CN105203663A/en active Pending
Non-Patent Citations (6)
| Title |
|---|
| HARILAOS DAMIANAKOS等: "Antimicrobial and Cytotoxic Isohexenylnaphthazarins from Arnebia euchroma(Royle) Jonst.(Boraginaceae) Callus and Cell Suspension Culture", 《MOLECULES》 * |
| SUKANDA TIANNIAM等: "Pyrolysis GC-MS-based metabolite fingerprinting for quality evaluation of commercial Angelica acutiloba roots", 《JOURNAL OF BIOSCIENCE AND BIOENGINEERING》 * |
| 丁毅等: "用裂解气相色谱-质谱法分析丁香油中的成分", 《中国生化药物杂志》 * |
| 严铸云 主编: "《药用植物与中药鉴定实验》", 30 September 2008 * |
| 刘大潜 主编: "《全国普通医药中专教材 中药制剂分析 》", 31 January 2000 * |
| 杨秀芳: "正交实验法优选紫草的提取工艺", 《中成药》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108918737A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | A kind of pre-treating method and efficient liquid-phase chromatography method measuring alkannin in w/o type Asian puccoon skin care milk |
| CN108918738A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | The pre-treating method and efficient liquid-phase chromatography method of alkannin content in a kind of measurement zicao soft plaster |
| CN108918739A (en) * | 2018-08-28 | 2018-11-30 | 华南协同创新研究院 | A kind of pre-treating method and efficient liquid-phase chromatography method measuring alkannin in O/W type Asian puccoon skin care milk |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Development of a chromatographic fingerprint for the chloroform extracts of Ganoderma lucidum by HPLC and LC–MS | |
| Zhang et al. | Simultaneous quantification of 17 constituents from Yuanhu Zhitong tablet using rapid resolution liquid chromatography coupled with a triple quadrupole electrospray tandem mass spectrometry | |
| Song et al. | Simultaneous determination of 19 flavonoids in commercial trollflowers by using high-performance liquid chromatography and classification of samples by hierarchical clustering analysis | |
| CN105203663A (en) | Rapid detection method for alkannin | |
| CN111089916B (en) | Method for detecting content of paeoniflorin, liquiritin and ammonium glycyrrhizinate in radix bupleuri and radix paeoniae alba oral liquid | |
| CN103163263B (en) | Method of analyzing composition and content of biomass pyrolysis products | |
| Gao et al. | A comprehensive strategy using chromatographic profiles combined with chemometric methods: Application to quality control of Polygonum cuspidatum Sieb. et Zucc | |
| CN105136935B (en) | A kind of quick determination method of Osthole | |
| Shi et al. | Simultaneous determination of eight components in Radix Tinosporae by high-performance liquid chromatography coupled with diode array detector and electrospray tandem mass spectrometry | |
| CN112114079A (en) | Method for simultaneously detecting 9 chemical components in quisqualis indica | |
| CN105136937A (en) | Magnolol and honokiol quick detection method | |
| CN104478686A (en) | Preparation method of ar-turmerone reference substance in turmeric volatile oil | |
| Zhao et al. | Gas chromatography–mass spectrometry for quantitative and qualitative analysis of essential oil from Curcuma wenyujin rhizomes | |
| CN104730193A (en) | Detection method of volatile component in heart stabilizing particles | |
| Zhou et al. | Fingerprint analysis of Daturae Flos using rapid resolution liquid chromatography-electrospray ionization mass spectrometry combined with stoichiometry | |
| Sun et al. | Combinative application of ph-zone-refining counter-current chromatography and preparative HPLC for the separation of alkaloids from Lycoris radiata | |
| CN105136936A (en) | Rapid detection method for psoralen and isopsoralen | |
| CN105203687A (en) | Paeonol quick detection method | |
| CN104267114A (en) | Detection method of nymphaea candida presl | |
| CN105203662A (en) | Method for rapidly detecting matrine | |
| CN105158362A (en) | Method for quickly detecting octadecanoic acid of whole blood erythrocytes | |
| CN105158387A (en) | Method for quickly detecting tetradecanoic acid of whole blood erythrocytes | |
| CN105203688A (en) | Fast detecting method for ligustilide | |
| CN105158396A (en) | Rapid detection method for tetramethylpyrazine | |
| CN105241968A (en) | Method of quickly detecting fatty acid in whole blood red cell |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151230 |