CN103837619A - Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof - Google Patents
Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof Download PDFInfo
- Publication number
- CN103837619A CN103837619A CN201410104187.XA CN201410104187A CN103837619A CN 103837619 A CN103837619 A CN 103837619A CN 201410104187 A CN201410104187 A CN 201410104187A CN 103837619 A CN103837619 A CN 103837619A
- Authority
- CN
- China
- Prior art keywords
- sophora
- tankinensis
- medicinal material
- fingerprint
- need testing
- 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.)
- Granted
Links
Images
Landscapes
- Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a fingerprint detection method for a traditional Chinese medicinal Ganyanlin injection and subprostrate sophora serving as a raw medicinal material thereof. The method comprises the steps of preparation of a test solution, construction of a contrast fingerprint and measurement of a test solution fingerprint. An optimal flowing phase constitution is screened out through a large amount of experiments; analysis conditions such as the wavelength, the chromatographic column and the column temperature are detected; multiple experiments show that the fingerprint detection method for the traditional Chinese medicinal Ganyanlin injection and the subprostrate sophora serving as the raw medicinal material thereof can comprehensively, objectively and accurately detect and evaluate the quality of the Ganyanlin injection, can guarantee the safety and the effectiveness of the raw medicinal material of the Ganyanlin injection from the source and has an important significance for guarantee of the clinical treating effect.
Description
Technical field
The present invention relates to a kind of quality detecting method of Chinese medicine preparation, be specifically related to the fingerprint atlas detection method of a kind of Sophora Tankinensis and raw medicinal material thereof.
Background technology
Sophora Tankinensis is through extracting the parenteral solution of being processed into by subprostrate sophora simple.Former recording in the 18 of " the Sanitation Ministry medicine standard " Traditional Chinese medicine historical preparation.The quality standard of Sophora Tankinensis has stipulated the check items such as the discriminating of alkaloid physics and chemistry, matrine thin-layer chromatography, pH value, residue on ignition at present, and adopt the content of titration measuring matrine, testing is loaded down with trivial details, detection efficiency is low, and the active component limited amount detecting, the well total quality of response preparation.Therefore, necessary on the basis of prior art, design research and development are a kind of can complete detection and the fingerprint atlas detection method of evaluating Sophora Tankinensis quality.Summary of the invention
Goal of the invention: the object of the invention is to solve the deficiencies in the prior art, the fingerprint atlas detection method of a kind of Sophora Tankinensis and raw medicinal material thereof is provided, this detection method can be evaluated the quality of Sophora Tankinensis objective, comprehensively and accurately, to controlling the quality of Sophora Tankinensis and the quality of raw medicinal material and guaranteeing that clinical efficacy is significant.
Technical scheme: in order to realize above technical purpose, the technical scheme that the present invention takes is:
The fingerprint atlas detection method of Chinese medicine Sophora Tankinensis, comprises the following steps:
(1) need testing solution preparation: by after Sophora Tankinensis dilution, with miillpore filter filtration, obtain need testing solution;
(2) reference fingerprint of high effective liquid chromatography for measuring Sophora Tankinensis: get 10 batches of above Sophora Tankinensis test samples sample introduction under suitable chromatographic condition, record the finger-print of each batch of Sophora Tankinensis, take these finger-prints as basis, calculate and obtain common pattern as Sophora Tankinensis reference fingerprint with similarity software;
(3) high effective liquid chromatography for measuring need testing solution finger-print: get step (1) Sophora Tankinensis need testing solution by the identical chromatographic condition sample introduction of step (2), record the finger-print of need testing solution;
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.
As preferred version, the fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis, in step (4), use 4 total fingerprint peakses of similarity software evaluation, concrete retention time is respectively No. 1 peak: 16.74min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min, and similarity is greater than 0.9.
As preferred version, the fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis, in step (1), need testing solution preparation method is: precision measures Sophora Tankinensis 1～2ml, put in 50～100ml measuring bottle, be diluted with water to scale, shake up, with the miillpore filter filtration of 0.45 μ m, to obtain final product.
As preferred version, the fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis, in step (2), chromatographic condition is: chromatographic column is take octadecylsilane chemically bonded silica as filling agent, mobile phase: 0.2%～01% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, column temperature is 30～40 ℃, and detection wavelength is 200～220nm.
As preferred version, the fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis, is characterized in that, in step (2), chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, mobile phase: 0.2% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, and 30 ℃ of column temperatures, detection wavelength is 205nm.
The fingerprint atlas detection method of Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora provided by the invention, comprises the following steps:
(1) need testing solution preparation: get subprostrate sophora medicinal material and suitably process, obtain need testing solution;
(2) reference fingerprint of Radix Sophorae Tonkinensis by HPLC medicinal material: get 10 batches of above subprostrate sophora medicinal material test samples sample introduction under suitable chromatographic condition, record the finger-print of each batch of subprostrate sophora medicinal material, take these finger-prints as basis, calculate and obtain common pattern as subprostrate sophora medicinal material reference fingerprint with similarity software;
(3) high effective liquid chromatography for measuring need testing solution finger-print: subprostrate sophora medicinal material need testing solution to be measured, by the identical chromatographic condition sample introduction of step (4), is recorded to the finger-print of need testing solution;
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.
The fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora, the described need testing solution preparation method of step (1) is: get subprostrate sophora medicinal powder (crossing sieve No. three) 0.5g, put in round-bottomed flask the precision 50ml that adds water, weighed weight, putting heating in electric jacket boils 4 hours, let cool, water is supplied the weight of less loss, after filtering with filter paper, with miillpore filter, (0.45 μ m) filters, and to obtain final product.
The fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora, the described chromatographic condition of step (2) and step (3) is: chromatographic column is take octadecylsilane chemically bonded silica as filling agent, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 30～40 ℃ of column temperatures, detection wavelength is 200～220nm.
As preferred version, the fingerprint atlas detection method of above-described Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora, the described chromatographic condition of step (2) and step (3) is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 30 ℃ of column temperatures, and detection wavelength is 205nm.
Chinese medicine Sophora Tankinensis fingerprint atlas detection method of the present invention is in the application of evaluating in Sophora Tankinensis end product quality.
Chinese medicine Sophora Tankinensis raw medicinal material fingerprint atlas detection method of the present invention is in the application of evaluating in Sophora Tankinensis raw medicinal material quality.
The present invention is by test sample finger-print and reference fingerprint machine similarity evaluation software (" the traditional Chinese medicine fingerprint area of computer aided similarity evaluation software 2004A version " of selecting Chinese Pharmacopoeia committee to recommend) calculating as calculated, obtain fingerprint similarity numerical value, according to similarity numerical value, be applied to the quality control of Sophora Tankinensis, should be greater than 0.90 as check quality control standard take fingerprint similarity; Be applied to the quality control of Sophora Tankinensis raw medicinal material subprostrate sophora, should be greater than 0.85 as check quality control standard take fingerprint similarity.The fingerprint atlas detection method of Sophora Tankinensis provided by the invention and raw medicinal material thereof is for the production of laboratory quality controls, and products obtained therefrom has higher quality stability, can guarantee that product safety is effective.
Adopt the Chinese medicine Sophora Tankinensis fingerprint atlas detection method that preferred version of the present invention provides to be tested with 4 total peaks, take No. 3 peaks as mark, have obvious absorption peak, for matrine compound, can be used as marker.
Adopt the Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora fingerprint atlas detection method that preferred version of the present invention provides to be tested with 5 total peaks, take No. 3 peaks as mark, have obvious absorption peak, for matrine compound, can be used as marker.
The fingerprint atlas detection method of Sophora Tankinensis raw medicinal material provided by the invention, select the testing conditions identical with Sophora Tankinensis finger print measuring method to carry out medicinal materials fingerprint investigation, be convenient to better investigate the correlativity of medicinal materials fingerprint and preparation finger, also be conducive to the difference in more each medicinal material place of production, for giving security in the raw medicinal material source of the safety and stability that guarantees Sophora Tankinensis.
The top condition screening process of Sophora Tankinensis provided by the invention and raw medicinal material fingerprint atlas detection method thereof:
1, instrument and reagent
Instrument: Agilent1100 liquid chromatograph.
Reagent: acetonitrile, tetrahydrofuran, chromatographically pure; It is pure that other reagent are analysis.
Reference substance: matrine, oxymatrine, provided by Nat'l Pharmaceutical & Biological Products Control Institute.
2, the screening of mobile phase kind
1. acetonitrile-absolute ethyl alcohol-water (80:10:10, phosphoric acid is adjusted pH2.0);
2. acetonitrile-0.2mol/L sodium dihydrogen phosphate (23:77);
3. methanol-water-triethylamine (50:50:0.05);
4. methanol-water-triethylamine (50:50:0.2);
5. acetonitrile-0.0257mol/L ammonium sulfate-SDS(40:60:0.6) sulfuric acid tune pH3.0
6. 0.2% lauryl sodium sulfate (10% phosphoric acid is adjusted pH3.0)-tetrahydrofuran (4:1)
Adopt above-mentioned mobile phase to measure, wherein 1., 2., 3. need on nh 2 column, measure, though can obtain good separating effect, retention time is unstable, calculates through software, and similarity result is undesirable; 4. mobile phase, is 5. the method that bibliographical information is measured matrine content, and through test, separating effect is undesirable, can not be by complete the alkaloids component separating in subprostrate sophora, and the accuracy of impact monitoring, and be not suitable for setting up finger-print.Therefore, the present invention gropes mobile phase composition repeatedly by great many of experiments, final definite 6. 0.2% lauryl sodium sulfate (10% phosphoric acid is adjusted pH3.0)-tetrahydrofuran (4:1) is mobile phase, adopt this mobile phase composition, each chromatographic peak separates fine, retention time is suitable, reaches finger-print information maximization principle, and reappearance to close stability excellent.
3, detect the selection of wavelength
Matrine reference substance solution in 200～400nm wavelength coverage without obvious absorption peaks, the comparative experiment shows, at 205～220nm place, especially at 205nm place, the each composition of Sophora Tankinensis has obvious absorption peak, therefore, the preferred 205nm of the present invention, as measuring wavelength, can obtain desirable chromatogram.
4, the selection of column temperature
The present invention analyzes Sophora Tankinensis respectively under 25 ℃, 30 ℃, 35 ℃ and 40 ℃ of column temperatures, contrast the chromatogram under each column temperature, experimental result shows, under 30-40 ℃ of column temperature condition, chromatographic peak can separate, under 30 ℃ of column temperature conditions, chromatographic peak separation case is best, post is pressed also lower, therefore, is optimum column temperature condition by 30 ℃.
5, the selection of chromatographic column
The present invention has screened different chromatographic columns by great many of experiments, adopts the chromatographic column of following model to measure:
1. nh 2 column: inertsil NH25u250mm × 4.6mmCat#:22871SN:3K150166
2. Alltech C
18post, 5 μ m, 4.6mm × 250mm
③SUPELCO
25cm×4.6mm，5μm，Col:50497154938-02
④CAPCELL?PAK?C18M.G5u250mm×4.6mmCat.No:90104Col.No.03169
⑤Agilent?ZORBAX*80A?Extend－C184.6×250mm5μ?PN:770450-902SN:USHR003183
⑥SUPELCO
25cm×4.6mm，5μm，Col:50497163280-05
⑦SUPELCO
25cm×4.6mm，5μm，Col:50497163486-01
Because this product effective constituent is biological alkali components, bibliographical information can adopt nh 2 column to measure its content.Through experiment, although nh 2 column can be obtained good separating effect, retention time is unstable, causes fingerprint similarity undesirable, therefore can not set up good finger-print and detect the quality of Sophora Tankinensis and raw medicinal material thereof.The present invention screens by great many of experiments, determines and adopts C
18post separates.6 C of four kinds of different manufacturers are investigated altogether
18the separating effect of chromatographic column, except separating effect 2., 4. is wherein undesirable, another four root chromatogram columns all can reach baseline separation by sample.Through chromatogram comparison, preferably adopt SUPELCO
25cm × 4.6mm, 5 μ m posts are as the analytical column of this product finger-print, and each chromatographic peak separates good, and reappearance and good stability, can set up stable finger-print.
6, the precision of finger-print and study on the stability
Get same Sophora Tankinensis need testing solution, sample introduction 6 times in 24 hours, to analyze for the first time gained finger-print as contrast, the similarity result of calculation of analyzing gained finger-print for all the other 5 times is respectively: 1.000,1.000,0.999,1.000,1.000,1.000, adopt above preferred chromatographic condition analysis, precision and the stability of the finger-print preparing are excellent.
7, reappearance is investigated
Get with 6 of a collection of Sophora Tankinensises, adopt above preferred chromatographic condition analysis, wherein once to analyze gained finger-print as contrast, analyze gained finger-print other 5 times and its similarity is respectively: 0.992,0.996,0.993,0.997,0.995; The fingerprint analysis method that shows to adopt the chromatographic condition that the present invention preferably obtains, has good reappearance.
Through above-mentioned great many of experiments screening, the optimum chromatogram condition that the present invention preferably obtains is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 30 ℃ of column temperatures, and detection wavelength is 205nm.
Beneficial effect: the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis provided by the invention and raw medicinal material thereof has the following advantages:
1, the many deficiencies that the present invention is directed to prior art detection method, filter out best mobile phase composition by great many of experiments, detect wavelength, chromatographic column, the analysis conditions such as column temperature, show that through many experiments checking detection method provided by the invention can be set up good finger-print.Adopt Chinese medicine Sophora Tankinensis fingerprint atlas detection method provided by the invention, can detect the various active composition in Chinese medicine Sophora Tankinensis simultaneously, thereby can be comprehensive, objective, the quality of detecting and assessing Sophora Tankinensis accurately, significant for guaranteeing clinical efficacy.
2, subprostrate sophora fingerprint atlas detection method provided by the invention, can Accurate Determining and evaluate the quality of subprostrate sophora medicinal material, for Sophora Tankinensis guarantees the safety of raw medicinal material from source, effectively, holds gate of the quality monitoring and has great importance.
Accompanying drawing explanation
Fig. 1 is Sophora Tankinensis reference fingerprint.
Fig. 2 is Sophora Tankinensis test sample finger-print.
Fig. 3 is subprostrate sophora medicinal material reference fingerprint.
Fig. 4 is subprostrate sophora medicinal material test sample finger-print.
Embodiment
Further illustrate the present invention below in conjunction with specific embodiment, should understand these embodiment is only not used in and limits the scope of the invention for the present invention is described, after having read the present invention, those skilled in the art all fall within the application's claims limited range to the modification of the various equivalent form of values of the present invention.
Foundation and the application of embodiment 1 Sophora Tankinensis reference fingerprint
1, the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis, comprises the following steps:
(1) need testing solution preparation method: precision measures Sophora Tankinensis 1ml, puts in 50ml measuring bottle, is diluted with water to scale, shakes up, and with miillpore filter, (0.45 μ m) filters, and to obtain final product.
(2) reference fingerprint of high effective liquid chromatography for measuring Sophora Tankinensis: the finger-print of getting 13 batches of Sophora Tankinensis test samples and record by following chromatographic condition each batch of Sophora Tankinensis, take these finger-prints as basis, calculate and obtain common pattern as Sophora Tankinensis reference fingerprint with similarity software.4 total fingerprint peakses in finger-print, detected altogether, concrete retention time (min) is respectively No. 1 peak: 16.74min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min.Reference fingerprint as shown in Figure 1.
Chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 30 ℃ of column temperatures, and detection wavelength is 205nm.
(3) high effective liquid chromatography for measuring need testing solution finger-print: get 13 batches by the Sophora Tankinensis need testing solution of step (1) gained by the identical chromatographic condition sample introduction of step (2), record the finger-print of need testing solution, as shown in Figure 2.
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.Specific experiment result is as shown in table 1:
Table 1 Sophora Tankinensis sample and similarity evaluation result
| Sequence number | Lot number | Similarity |
| 1 | 120609 | 0.986 |
| 2 | 120910 | 0.985 |
| 3 | 120613 | 0.986 |
| 4 | 120915 | 0.988 |
| 5 | 121022 | 0.993 |
| 6 | 121021 | 0.994 |
| 7 | 121229 | 0.990 |
| 8 | 121231 | 0.995 |
| 9 | 130112 | 0.991 |
| 10 | 130114 | 0.993 |
| 11 | 130116 | 0.993 |
| 12 | 130103 | 0.998 |
| 13 | 130402 | 0.996 |
Result of calculation by above table 1 shows, each batch of Sophora Tankinensis fingerprint similarity is higher, is all greater than 0.90, shows that production technology is more stable, and quality homogeneity is good.Also prove simultaneously, adopt fingerprint atlas detection method provided by the invention can objectively reflect the quality of Sophora Tankinensis, for guaranteeing that clinical safety curative effect has great importance comprehensively.
Foundation and the application of the reference fingerprint of embodiment 2 Sophora Tankinensis raw medicinal material subprostrate sophoras
(1) need testing solution preparation: 10 batches of subprostrate sophora medicinal powders (the crossing sieve No. three) 0.5g that gets respectively the place of production, Guangxi, put in round-bottomed flask, the precision 50ml that adds water, weighed weight, puts heating in electric jacket and boils 4 hours, lets cool, water is supplied the weight of less loss, after filtering with filter paper, with miillpore filter, (0.45 μ m) filters, and to obtain final product.
(2) reference fingerprint of Radix Sophorae Tonkinensis by HPLC medicinal material: get 10 batches of subprostrate sophora medicinal material need testing solutions that step (1) prepares, record the finger-print of every batch of subprostrate sophora medicinal material by following chromatographic condition, take these finger-prints as basis, calculate and obtain common pattern finger-print in contrast with similarity software.4 total fingerprint peakses in finger-print, detected altogether, concrete retention time (min) is respectively: No. 1 peak: 16.7min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min, No. 5 peak: 35.4min.As shown in Figure 3.
Chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 30 ℃ of column temperatures, and detection wavelength is 205nm.
(3) Radix Sophorae Tonkinensis by HPLC medicinal material need testing solution finger-print: get 10 batches of need testing solutions of step (1) by the identical chromatographic condition sample introduction of step (2), record the finger-print of subprostrate sophora medicinal material need testing solution, as shown in Figure 4.
(4) get the subprostrate sophora medicinal material reference fingerprint similarity software evaluation that subprostrate sophora medicinal material need testing solution finger-print that step (3) obtains and step (2) obtain, the similarity of calculating respectively each batch of subprostrate sophora medicinal materials fingerprint and reference fingerprint comparison, the results are shown in Table 2.Result shows that the fingerprint similarity of different batches subprostrate sophora medicinal material is all greater than 0.90, show that subprostrate sophora medicinal material reference fingerprint that the present invention sets up can be used for detecting and controlling the quality of the raw medicinal material subprostrate sophora of Sophora Tankinensis, for guaranteeing the safety of raw medicinal material, effectively, holding gate of the quality monitoring has great importance.
Table 2 subprostrate sophora medicinal materials fingerprint similarity evaluation result
| Lot number | Similarity | Lot number | Similarity |
| 120418 | 0.988 | 120912 | 0.923 |
| 120510 | 0.936 | 121008 | 0.910 |
| 120516 | 0.901 | 121102 | 0.973 |
| 120730 | 0.902 | 121123 | 0.915 |
| 120829 | 0.901 | 130429 | 0.905 |
Foundation and the application of embodiment 3 Sophora Tankinensis reference fingerprints
1, the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis, comprises the following steps:
(1) need testing solution preparation method: precision measures Sophora Tankinensis 2ml, puts in 100ml measuring bottle, is diluted with water to scale, shakes up, and with miillpore filter, (0.45 μ m) filters, and to obtain final product.
(2) reference fingerprint of high effective liquid chromatography for measuring Sophora Tankinensis: the finger-print of getting 13 batches of Sophora Tankinensis test samples and record by following chromatographic condition each batch of Sophora Tankinensis, take these finger-prints as basis, calculate and obtain common pattern as Sophora Tankinensis reference fingerprint with similarity software.4 total fingerprint peakses in finger-print, detected altogether, concrete retention time (min) is respectively No. 1 peak: 16.75min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min.
Chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 40 ℃ of column temperatures, and detection wavelength is 220nm.
(3) high effective liquid chromatography for measuring need testing solution finger-print: get 13 batches by the Sophora Tankinensis need testing solution of step (1) gained by the identical chromatographic condition sample introduction of step (2), record the finger-print of need testing solution.
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.Specific experiment result is as shown in table 3:
Table 3 Sophora Tankinensis sample and similarity evaluation result
| Sequence number | Lot number | Similarity |
| 1 | 120609 | 0.985 |
| 2 | 120910 | 0.985 |
| 3 | 120613 | 0.986 |
| 4 | 120915 | 0.987 |
| 5 | 121022 | 0.992 |
| 6 | 121021 | 0.993 |
| 7 | 121229 | 0.992 |
| 8 | 121231 | 0.994 |
| 9 | 130112 | 0.992 |
| 10 | 130114 | 0.992 |
| 11 | 130116 | 0.993 |
| 12 | 130103 | 0.995 |
| 13 | 130402 | 0.995 |
Result of calculation by above table 3 shows, each batch of Sophora Tankinensis fingerprint similarity is higher, is all greater than 0.90, shows that production technology is more stable, and quality homogeneity is good.Also prove simultaneously, adopt fingerprint atlas detection method provided by the invention can objectively reflect the quality of Sophora Tankinensis, for guaranteeing that clinical safety curative effect has great importance comprehensively.
Foundation and the application of the reference fingerprint of embodiment 4 Sophora Tankinensis raw medicinal material subprostrate sophoras
(1) need testing solution preparation: 10 batches of subprostrate sophora medicinal powders (the crossing sieve No. three) 1g that gets respectively the place of production, Guangxi, put in round-bottomed flask, the precision 100ml that adds water, weighed weight, puts heating in electric jacket and boils 2 hours, lets cool, water is supplied the weight of less loss, after filtering with filter paper, with miillpore filter, (0.45 μ m) filters, and to obtain final product.
(2) reference fingerprint of Radix Sophorae Tonkinensis by HPLC medicinal material: get 10 batches of subprostrate sophora medicinal material need testing solutions that step (1) prepares, record the finger-print of every batch of subprostrate sophora medicinal material by following chromatographic condition, take these finger-prints as basis, calculate and obtain common pattern finger-print in contrast with similarity software.4 total fingerprint peakses in finger-print, detected altogether, concrete retention time (min) is respectively: No. 1 peak: 16.7min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min, No. 5 peak: 35.4min.
Chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, 0.2% lauryl sodium sulfate take volume ratio as 4:1: tetrahydrofuran is mobile phase, 40 ℃ of column temperatures, and detection wavelength is 220nm.
(3) Radix Sophorae Tonkinensis by HPLC medicinal material need testing solution finger-print: get 10 batches of need testing solutions of step (1) by the identical chromatographic condition sample introduction of step (2), record the finger-print of subprostrate sophora medicinal material need testing solution.
(4) get the subprostrate sophora medicinal material reference fingerprint similarity software evaluation that subprostrate sophora medicinal material need testing solution finger-print that step (3) obtains and step (2) obtain, the similarity of calculating respectively each batch of subprostrate sophora medicinal materials fingerprint and reference fingerprint comparison, the results are shown in Table 4.Result shows that the fingerprint similarity of different batches subprostrate sophora medicinal material is all greater than 0.90, show that subprostrate sophora medicinal material reference fingerprint that the present invention sets up can be used for detecting and controlling the quality of the raw medicinal material subprostrate sophora of Sophora Tankinensis, for guaranteeing the safety of raw medicinal material, effectively, holding gate of the quality monitoring has great importance.
Table 4 subprostrate sophora medicinal materials fingerprint similarity evaluation result
| Lot number | Similarity | Lot number | Similarity |
| 120418 | 0.988 | 120912 | 0.925 |
| 120510 | 0.936 | 121008 | 0.914 |
| 120516 | 0.901 | 121102 | 0.983 |
| 120730 | 0.902 | 121123 | 0.955 |
| 120829 | 0.901 | 130429 | 0.925 |
Show by above experimental result, the fingerprint atlas detection method of Sophora Tankinensis provided by the invention and raw medicinal material thereof, can be objective, comprehensively, analyze targetedly and evaluate the quality of Sophora Tankinensis finished product and raw medicinal material, significant to guaranteeing clinical efficacy, fingerprint atlas detection method provided by the invention, can be simultaneously for analyzing raw medicinal material and finished product Sophora Tankinensis preparation, compared with a kind of composition in the single analysis raw medicinal material of prior art or Sophora Tankinensis, there is obvious technical progress.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis, is characterized in that, comprises the following steps:
(1) need testing solution preparation: by after Sophora Tankinensis dilution, with miillpore filter filtration, obtain need testing solution;
(2) reference fingerprint of high effective liquid chromatography for measuring Sophora Tankinensis: get 10 batches of above Sophora Tankinensis test samples sample introduction under suitable chromatographic condition, record the finger-print of each batch of Sophora Tankinensis, take these finger-prints as basis, calculate and obtain common pattern as Sophora Tankinensis reference fingerprint with similarity software;
(3) high effective liquid chromatography for measuring need testing solution finger-print: get step (1) Sophora Tankinensis need testing solution by the identical chromatographic condition sample introduction of step (2), record the finger-print of need testing solution;
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.
2. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis according to claim 1, it is characterized in that, in step (4), use 4 total fingerprint peakses of similarity software evaluation, concrete retention time is respectively No. 1 peak: 16.74min, No. 2 peak: 28.3min, No. 3 peak: 29.9min, No. 4 peak: 32.9min, and similarity is greater than 0.9.
3. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis according to claim 1, it is characterized in that, in step (1), need testing solution preparation method is: precision measures Sophora Tankinensis 1～2ml, put in 50～100ml measuring bottle, be diluted with water to scale, shake up, with the miillpore filter filtration of 0.45 μ m, to obtain final product.
4. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis according to claim 3, it is characterized in that, in step (1), need testing solution preparation method is: precision measures Sophora Tankinensis 1ml, put in 50ml measuring bottle, be diluted with water to scale, shake up, with the miillpore filter filtration of 0.45 μ m, to obtain final product.
5. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis according to claim 1, it is characterized in that, in step (2), chromatographic condition is: chromatographic column is take octadecylsilane chemically bonded silica as filling agent, mobile phase: 0.2%～01% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, column temperature is 30～40 ℃, and detection wavelength is 200～220nm.
6. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis according to claim 5, is characterized in that, in step (2), chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, mobile phase: 0.2% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, and 30 ℃ of column temperatures, detection wavelength is 205nm.
7. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora, is characterized in that, comprises the following steps:
(1) need testing solution preparation: get subprostrate sophora medicinal material and suitably process, obtain need testing solution;
(2) reference fingerprint of Radix Sophorae Tonkinensis by HPLC medicinal material: get 10 batches of above subprostrate sophora medicinal material test samples sample introduction under suitable chromatographic condition, record the finger-print of each batch of subprostrate sophora medicinal material, take these finger-prints as basis, calculate and obtain common pattern as subprostrate sophora medicinal material reference fingerprint with similarity software;
(3) high effective liquid chromatography for measuring need testing solution finger-print: get step (1) subprostrate sophora medicinal material need testing solution by the identical chromatographic condition sample introduction of step (2), record the finger-print of need testing solution;
(4) the reference fingerprint similarity software evaluation that need testing solution finger-print step (3) being obtained and step (2) obtain.
8. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora according to claim 7, it is characterized in that, in step (1), need testing solution preparation method is: get subprostrate sophora medicinal powder 0.5～1g, put in round-bottomed flask, precision 50～the 100ml that adds water, weighed weight, puts heating in electric jacket and boils 1～4 hour, lets cool, water is supplied the weight of less loss, after filtering with filter paper, then with 0.45 μ m miillpore filter filtration, to obtain final product.
9. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora according to claim 7, it is characterized in that, in step (2), chromatographic condition is: chromatographic column is take octadecylsilane chemically bonded silica as filling agent, mobile phase: 0.2～1%% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, 30～40 ℃ of column temperatures, detection wavelength is 200～220nm.
10. the fingerprint atlas detection method of Chinese medicine Sophora Tankinensis raw medicinal material subprostrate sophora according to claim 7, is characterized in that, in step (2), chromatographic condition is: take specification as 5 μ m, SUPELCO
25cm × 4.6mm is chromatographic column, mobile phase: 0.2% lauryl sodium sulfate that volume ratio is 4:1: tetrahydrofuran, and 30 ℃ of column temperatures, detection wavelength is 205nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410104187.XA CN103837619B (en) | 2014-03-20 | 2014-03-20 | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410104187.XA CN103837619B (en) | 2014-03-20 | 2014-03-20 | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103837619A true CN103837619A (en) | 2014-06-04 |
| CN103837619B CN103837619B (en) | 2015-06-17 |
Family
ID=50801333
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410104187.XA Active CN103837619B (en) | 2014-03-20 | 2014-03-20 | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103837619B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107037146A (en) * | 2017-02-17 | 2017-08-11 | 丹东药业集团有限公司 | A kind of method of the wooden chicken granular mass control of compound |
| CN109030694A (en) * | 2018-09-20 | 2018-12-18 | 广西壮族自治区药用植物园 | The method of the finger-print identification subprostrate sophora true and false |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101458235A (en) * | 2008-12-30 | 2009-06-17 | 上海纳米技术及应用国家工程研究中心有限公司 | Matrine liquid chromatography measuring method |
| CN102000140A (en) * | 2010-11-08 | 2011-04-06 | 承德燕峰药业有限责任公司 | Novel method for synchronously quantifying matrine and oxymatrine in kuh-seng and preparations of kuh-seng |
| CN103630630A (en) * | 2013-12-16 | 2014-03-12 | 广西大学 | Method for measuring content of matrine in medicinal material of subprostrate sophora |
-
2014
- 2014-03-20 CN CN201410104187.XA patent/CN103837619B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101458235A (en) * | 2008-12-30 | 2009-06-17 | 上海纳米技术及应用国家工程研究中心有限公司 | Matrine liquid chromatography measuring method |
| CN102000140A (en) * | 2010-11-08 | 2011-04-06 | 承德燕峰药业有限责任公司 | Novel method for synchronously quantifying matrine and oxymatrine in kuh-seng and preparations of kuh-seng |
| CN103630630A (en) * | 2013-12-16 | 2014-03-12 | 广西大学 | Method for measuring content of matrine in medicinal material of subprostrate sophora |
Non-Patent Citations (5)
| Title |
|---|
| 国家药典委员会: "《卫生部药品标—中成药成方制剂第十八册》", 31 January 2001, article "肝炎灵注射液", pages: 139 * |
| 李妃，李成平，付晖，粟晓黎，王兆基，侯晓蓉: "山豆根研究进展及毒性成分检测方法补充报道", 《药物分析杂志》, vol. 33, no. 8, 31 August 2013 (2013-08-31), pages 1453 - 1463 * |
| 邓富良 等: "H P L C 法测定肝炎灵注射液中苦参碱的含量", 《中药新药与临床药理》, vol. 14, no. 6, 30 November 2003 (2003-11-30), pages 405 - 407 * |
| 黄亚非，黄际薇，陶玲，麦海燕，张永明: "广西不同产地山豆根的指纹图谱特征研究", 《中药材》, vol. 28, no. 1, 31 January 2005 (2005-01-31), pages 21 - 22 * |
| 黄颖，王乃平，陈勇: "广西产山豆根HPLC指纹图谱测定", 《中国实验方剂学杂志》, vol. 17, no. 14, 31 July 2011 (2011-07-31), pages 66 - 68 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107037146A (en) * | 2017-02-17 | 2017-08-11 | 丹东药业集团有限公司 | A kind of method of the wooden chicken granular mass control of compound |
| CN109030694A (en) * | 2018-09-20 | 2018-12-18 | 广西壮族自治区药用植物园 | The method of the finger-print identification subprostrate sophora true and false |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103837619B (en) | 2015-06-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104181248B (en) | A kind of fingerprint atlas detection method of Ganmaoling Granules | |
| CN103698413B (en) | The method for building up of Cape jasmine and Magnolia Decoction particle HPLC finger-print and standard finger-print thereof | |
| CN106525989A (en) | Detection method of fructus aurantii medicine material fingerprint and standard fingerprint | |
| CN104569252B (en) | A kind of method for building up of the finger printing of Chinese medicine composition | |
| CN108226321A (en) | The fingerprint atlas detection method and its finger-print of Bi bells stomachache particle | |
| CN107356691A (en) | Build the detection method of bent finger-print | |
| CN103115892A (en) | Method for preparing ginkgo leaf extract by using near infrared spectroscopy analysis technology | |
| CN105675739A (en) | Construction method of HPLC specific chromatogram of traditional Chinese medicines treating wind-heat cold | |
| CN105675744B (en) | The fingerprint atlas detection method of blue or green money willow | |
| CN102539597B (en) | Method for quickly identifying notopterygium incisum seed and notopterygium franchetii seed | |
| CN102441057B (en) | High performance liquid chromatography (HPLC) fingerprint detection method for blood-nourishing brain-refreshing grain | |
| CN103837619B (en) | Fingerprint detection method for traditional Chinese medicinal Ganyanlin injection and raw medicinal material thereof | |
| CN111175429A (en) | Method for establishing fingerprint spectrum of bactericidal and antipruritic lotion | |
| CN104849375B (en) | The detection method of 'Juhong Tanke ' | |
| CN102435689A (en) | Determination method of UPLC-MS (ultrahigh performance liquid chromatography-mass spectrometry) fingerprint of Radix Scutellariae medicinal material | |
| CN103472148A (en) | Fingerprint spectrum detection method of Chinese medicine composition preparation | |
| CN101013118A (en) | Method for constructing HPLC standard fingerprint pattern of dog-ridge medicinal materials and quality identification | |
| CN102106950A (en) | Quality control method in NuJin capsule extraction and concentration process | |
| CN107576739A (en) | A kind of HPLC fingerprint atlas detection methods of LONGMU ZHUANGGU KELI | |
| CN104655748B (en) | A kind of erigeron breviscapus granule finger-print and its method for building up and application | |
| CN102890125A (en) | Building method and mass detection method for fingerprint of total alkaloid components of cortex mori radicis medicinal material or cortex mori radicis extract | |
| CN104807932A (en) | Determining method of fingerprint spectrum of flavonoid ingredient of fenugreek seed extract | |
| CN203479772U (en) | High performance liquid chromatography fingerprint measuring device of dendrobium candidum | |
| CN102507830B (en) | High performance liquid chromatograph method for measuring content of Quzhazhigan in Rheum lhasaense | |
| CN113759057B (en) | Characteristic spectrum of allium macrostemon white water extract and preparation thereof and construction method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |