CN102650626B - Standard fingerprint of fleabane phenol bulk drug and preparation, establishment method and application - Google Patents
Standard fingerprint of fleabane phenol bulk drug and preparation, establishment method and application Download PDFInfo
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Abstract
The invention provides a standard fingerprint of fleabane phenol bulk drugs and preparations, an establishment method and application. The technical schemes comprise the determination of the standard fingerprint and the characteristic fingerprint peak of fleabane drug materials, the fleabane phenol bulk drugs and the preparations, and the application of the standard fingerprint in the aspects of extraction and processing, and quality control in the process of producing the fleabane phenol bulk drugs and the preparations. Therefore, the overall monitoring of the process and the quality from the fleabane drug materials to fleabane phenol bulk drugs and then the preparations can be realized, basic substances of the fleabane drug materials, the fleabane phenol bulk drugs and the preparations can be conveniently confirmed, and the production process and quality can be conveniently evaluated, and standard indications can be provided for subsequent process improvement and perfection.
Description
Technical field
The present invention relates to a kind of Chinese medicine from the medicinal material to the bulk drug again to the method for building up of standard preparation finger-print, relate in particular to a kind of erigeron breviscapus bulk drug and standard preparation finger-print and method for building up thereof, and by the standard finger-print of the method acquisition and in the application aspect bulk drug and preparation process quality control.
Background technology
That traditional Chinese medicine fingerprint refers to is common in certain Chinese crude drug or its bulk drug, preparation, have distinctive certain class or the chromatogram of number constituents or the collection of illustrative plates of spectrum.In the situation that in present stage, the effective elements of the medicine is most very not clear and definite, the foundation of finger-print has great importance for the quality of effective control Chinese crude drug and bulk drug thereof, preparation.
At present, the research of the finger-print of relevant erigeron breviscapus medicinal material has been reported.As Xie Peishan chief editor " chromatographic fingerprints of Chinese materia medica " (People's Health Publisher, 2005 February the 1st edition) book, the HPLC chromatographic fingerprinting research of erigeron breviscapus medicinal material is elaborated.By 48 batches of samples from the various places, Yunnan are carried out the HPLC fingerprint map analyzing, set up the HPLC finger-print of erigeron breviscapus medicinal material in book, formed the common pattern of finger-print, for the quality analysis of erigeron breviscapus medicinal material is had laid a good foundation.The described finger-print of this research has 21 characteristic fingerprint peaks, but it has only pointed out wherein 14, and the rate of pointing out only reaches 67%, has certain defect on the basic substance understanding to the erigeron breviscapus medicinal material; And it only carries out for the erigeron breviscapus medicinal material, does not relate to the deep processing product of erigeron breviscapus, so that its range of application is restricted, and can not well carry out technique and quality control in erigeron breviscapus bulk drug and preparation production run.
Summary of the invention
therefore, the objective of the invention is for the finger-print that the erigeron breviscapus medicinal material is only arranged at present and rate lower (67%) is pointed out at characteristic fingerprint peak in this collection of illustrative plates, can not realize well the technique from the erigeron breviscapus medicinal material to erigeron breviscapus bulk drug and preparation, the deficiency of quality, provide a kind of rate of pointing out to reach 95% erigeron breviscapus medicinal materials fingerprint, and the method for building up of the finger-print of erigeron breviscapus bulk drug and preparation and the standard finger-print of acquisition, thereby realize the technique from the erigeron breviscapus medicinal material to erigeron breviscapus bulk drug and preparation, the quality whole process supervision, be convenient to the erigeron breviscapus medicinal material, erigeron breviscapus bulk drug and preparation basic substance are confirmed, production technology and quality good or not are made an appraisal, and be that follow-up process modification provides the standard indication with improving.
For above-mentioned purpose, technical scheme of the present invention is as follows:
On the one hand, the invention provides the method for building up that a kind of erigeron breviscapus medicinal material and erigeron breviscapus bulk drug arrive the standard finger-print of preparation again, comprise the following steps: get scutellarin and be dissolved in right amount in methyl alcohol as object of reference solution; Get respectively again erigeron breviscapus medicinal material, erigeron breviscapus bulk drug and preparation and make need testing solution; Then get with the high performance liquid chromatograph test respectively.
Preferably, the DAD detecting device is adopted in described high performance liquid chromatograph test, detects in wavelength 270nm and 330nm.
Preferably, after described erigeron breviscapus medicinal material extracts with methyl alcohol, dissolve with methyl alcohol, filter, obtain erigeron breviscapus medicinal material need testing solution, obtain the standard finger-print of erigeron breviscapus medicinal material with the high performance liquid chromatograph test, preferably, described methanol solution for dissolving is that volume ratio is 60% methanol solution again.
preferably, contain 21 principal character fingerprint peakses in the standard finger-print of described erigeron breviscapus medicinal material, the peak is arranged No. 1: the pyromeconic acid glucoside, No. 2 peaks: pyromeconic acid, No. 3 peaks: protocatechuic acid, No. 4 peaks: chlorogenic acid+P-hydroxybenzoic acid, No. 5 peaks: the unknown, No. 6 peaks: caffeic acid+multiradiate fleabane glycosides, No. 7 peaks: syringic acid, No. 8 peaks: fleabane flower glycosides, No. 9 peaks: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 16 peaks: 6-hydroxyl Kaempferol, No. 17 peaks: high baicalein, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.Wherein, No. 1 peak: pyromeconic acid glucoside, No. 8 peaks: fleabane flower glycosides, No. 16 peaks: 6-hydroxyl Kaempferol, No. 18 peaks: 1; 3; 5-three caffeoyl quinic acids, No. 19 peaks: 3; 4; 5-three caffeoyl quinic acids, No. 21 peaks: the Erigeroster B ethyl ester, be 6 basic substances that the present invention further points out out on the HPLC chromatographic fingerprinting Research foundation of Xie Peishan to the erigeron breviscapus medicinal material, erigeron breviscapus medicinal material standard finger-print characteristic fingerprint peak is pointed out rate and is reached 95%.
Preferably, the standard finger-print of described erigeron breviscapus medicinal material as shown in Figure 1a.
Preferably, described erigeron breviscapus bulk drug is dissolved in methyl alcohol, obtains erigeron breviscapus bulk drug need testing solution, then obtains the standard finger-print of erigeron breviscapus bulk drug with the high performance liquid chromatograph test.
preferably, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus bulk drug, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
Preferably, in the standard finger-print of described erigeron breviscapus bulk drug, the number percent of the shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 39.79%, No. 10 peaks: 0.95%, No. 11 peaks: 32.39%, No. 12 peaks: 5.90%, No. 13 peaks: 6.48%, No. 14 peaks: 1.51%, No. 15 peaks: 8.16%, No. 18 peaks: 0.55%, No. 19 peaks: 0.33%, No. 20 peaks: 1.30% and No. 21 peak: 0.42%.
Preferably, the standard finger-print of described erigeron breviscapus bulk drug is as shown in Fig. 2 a.
Preferably, described erigeron breviscapus preparation is dissolved in methyl alcohol, obtains erigeron breviscapus preparation need testing solution, then obtains the standard finger-print of erigeron breviscapus preparation with the high performance liquid chromatograph test.
preferably, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus preparation, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, and identical with 11 characteristic fingerprint peaks in the erigeron breviscapus bulk drug, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
Preferably, in the standard finger-print of described erigeron breviscapus preparation, the number percent of the shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 37.10%, No. 10 peaks: 1.68%, No. 11 peaks: 26.75%, No. 12 peaks: 8.20%, No. 13 peaks: 6.44%, No. 14 peaks: 1.31%, No. 15 peaks: 8.65%, No. 18 peaks: 1.26%, No. 19 peaks: 0.83%, No. 20 peaks: 3.15% and No. 21 peak: 0.58%.
Preferably, the standard finger-print of described erigeron breviscapus preparation is as shown in Fig. 3 a.
On the other hand, the invention provides a kind of erigeron breviscapus medicinal material that is made by said method and the standard finger-print of erigeron breviscapus bulk drug and preparation.
preferably, contain 21 principal character fingerprint peakses in the standard finger-print of described erigeron breviscapus medicinal material, the peak is arranged No. 1: the pyromeconic acid glucoside, No. 2 peaks: pyromeconic acid, No. 3 peaks: protocatechuic acid, No. 4 peaks: chlorogenic acid+P-hydroxybenzoic acid, No. 5 peaks: the unknown, No. 6 peaks: caffeic acid+multiradiate fleabane glycosides, No. 7 peaks: syringic acid, No. 8 peaks: fleabane flower glycosides, No. 9 peaks: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 16 peaks: 6-hydroxyl Kaempferol, No. 17 peaks: high baicalein, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.Wherein, No. 1 peak: pyromeconic acid glucoside, No. 8 peaks: fleabane flower glycosides, No. 16 peaks: 6-hydroxyl Kaempferol, No. 18 peaks: 1; 3; 5-three caffeoyl quinic acids, No. 19 peaks: 3; 4; 5-three caffeoyl quinic acids, No. 21 peaks: Erigeroster B ethyl ester, 6 basic substances further pointing out out on the HPLC chromatographic fingerprinting Research foundation of Xie Peishan to the erigeron breviscapus medicinal material for the present invention.
Preferably, the standard finger-print of described erigeron breviscapus medicinal material as shown in Figure 1a.
preferably, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus bulk drug, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
Preferably, in the standard finger-print of described erigeron breviscapus bulk drug, the number percent of the total shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 39.79%, No. 10 peaks: 0.95%, No. 11 peaks: 32.39%, No. 12 peaks: 5.90%, No. 13 peaks: 6.48%, No. 14 peaks: 1.51%, No. 15 peaks: 8.16%, No. 18 peaks: 0.55%, No. 19 peaks: 0.33%, No. 20 peaks: 1.30% and No. 21 peak: 0.42%.
Preferably, the standard finger-print of described erigeron breviscapus bulk drug is as shown in Fig. 2 a.
preferably, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus preparation, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, and identical with 11 characteristic fingerprint peaks in the erigeron breviscapus bulk drug, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
Preferably, in the standard finger-print of described erigeron breviscapus preparation, the number percent of the total shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 37.10%, No. 10 peaks: 1.68%, No. 11 peaks: 26.75%, No. 12 peaks: 8.20%, No. 13 peaks: 6.44%, No. 14 peaks: 1.31%, No. 15 peaks: 8.65%, No. 18 peaks: 1.26%, No. 19 peaks: 0.83%, No. 20 peaks: 3.15% and No. 21 peak: 0.58%.
Preferably, the standard finger-print of described erigeron breviscapus preparation is as shown in Fig. 3 a.
Again on the one hand, the invention provides a kind of from the erigeron breviscapus medicinal material to the erigeron breviscapus bulk drug again to the standard finger-print of preparation, and produce feed intake application aspect screening, extraction processing technology and quality control of Chinese crude drug at erigeron breviscapus bulk drug and preparation.
Erigeron breviscapus medicinal material standard finger-print of the present invention, its 21 characteristic fingerprint peaks have comprised (No. 9 peaks: scutellarin, No. 11 peaks: 3,5 active component peaks clear and definite in erigeron breviscapus bulk drug and preparation, 5-DCQ, No. 12 peaks: 1,5-DCQ, No. 13 peaks: Erigeroster B, No. 15 peaks: 4, the 5-DCQ), different batches medicinal materials fingerprint similarity 〉=0.9 can feed intake; From the erigeron breviscapus medicinal material to the erigeron breviscapus bulk drug again to the standard preparation finger-print, erigeron breviscapus bulk drug and formulation characteristics fingerprint peaks all derive from medicinal material, extraction and production run are not introduced other impurity, show clear and definite sibship and high correlativity; The erigeron breviscapus bulk drug extracts and during preparation production, and in medicinal material, the 1-8 impurity peaks is all removed, 9-15 active component peak all keeps, 16-21 peak clearance is greater than 90.0%, has guaranteed bulk drug and the quality of the pharmaceutical preparations and pharmacologically active; Erigeron breviscapus bulk drug and standard preparation Fingerprints fingerprint peaks and peak area thereof have been determined contained 11 basic substances of erigeron breviscapus and each basic substance ratio, are convenient to bulk drug and the quality of the pharmaceutical preparations and control, and guarantee its stability, homogeneity.
Description of drawings
Below, describe by reference to the accompanying drawings embodiment of the present invention in detail, wherein:
Fig. 1 is the standard finger-print of erigeron breviscapus medicinal material and the collection of illustrative plates of 21 batches of erigeron breviscapus medicinal materials fingerprint comparisons, picture in picture 1a is the standard finger-print of erigeron breviscapus medicinal material, and Fig. 1 b is 21 crowdes of erigeron breviscapus medicinal materials fingerprint similarity evaluation stacking diagrams (getting median);
Fig. 2 is the standard finger-print of erigeron breviscapus bulk drug and the collection of illustrative plates of 10 batches of erigeron breviscapus bulk drug finger-print comparisons, picture in picture 2a is the standard finger-print of erigeron breviscapus bulk drug, and Fig. 2 b is 10 crowdes of erigeron breviscapus bulk drug fingerprint similarity evaluation stacking diagrams (getting median);
Fig. 3 is the standard finger-print of erigeron breviscapus preparation and the collection of illustrative plates of 10 batches of erigeron breviscapus preparation finger comparisons, picture in picture 3a is the standard finger-print of erigeron breviscapus preparation, and Fig. 3 b is 10 crowdes of erigeron breviscapus preparation finger similarity evaluation stacking diagrams (getting median).
Embodiment
Embodiment 1: erigeron breviscapus medicinal material standard finger-print is set up
1, chromatographic condition and system suitability Agilent 1100 Series high performance liquid chromatographs; Zorbax SB-C18 (4.6mm * 250mm, 5 μ m) chromatographic column; Take acetonitrile-methyl alcohol-formic acid as mobile phase; Detect wavelength: 270nm, 330nm; Theoretical cam curve should be not less than 6000 by scutellarin peak calculating.
2, it is appropriate that the scutellarin reference substance is got in the preparation of object of reference solution, is mixed with the methanol solution of 0.4mg/mL, and get final product.
3, erigeron breviscapus medicinal material 0.50g is got in the preparation of need testing solution, uses the methyl alcohol heating and refluxing extraction, dissolves with 60% methyl alcohol after the extract evaporate to dryness and is settled to 10mL, through 0.45 μ m membrane filtration, and get final product.
4, accurate object of reference solution and each 5 μ L of need testing solution of drawing of determination method difference, the injection liquid chromatography is measured and get final product.
5, erigeron breviscapus medicinal material standard finger-print is set up
According to 42 test datas of 21 batch sample; erigeron breviscapus medicinal material standard finger-print has 21 principal character fingerprint peakses; has the Fingerprints shown in Fig. 1 a and table 1-1; the rate of pointing out of this collection of illustrative plates reaches 95%; wherein comprise erigeron breviscapus raw material and preparation clear and definite (No. 9 peaks: scutellarin, No. 11 peaks: 3,5 active component peaks; 5-DCQ, No. 12 peaks: 1; 5-DCQ, No. 13 peaks: Erigeroster B, No. 15 peaks: 4,5-DCQ)." similarity evaluation " that different batches erigeron breviscapus medicinal materials fingerprint and erigeron breviscapus medicinal material standard finger-print adopt Chinese Pharmacopoeia Commission to recommend (2004B) calculates, and similarity answers 〉=0.9.
Table 1-1 erigeron breviscapus medicinal material principal character fingerprint peaks table
6,21 batches of erigeron breviscapus medicinal material sample finger-prints
Gather 21 batches, erigeron breviscapus medicinal material sample, make respectively need testing solution, 42 groups of data are analyzed in the every batch of replicate determination 2 times, the finger-print that records of observation post directly perceived, and overall picture is without significant change; Calculate with similarity respectively, the finger-print and the similarity of its gained common pattern collection of illustrative plates that record on instrument are on the same stage pressed median calculating, the 20 batches of medicinal material similarities greater than 0.9,1 batch of medicinal material similarity between 0.8-0.9; Press average and calculate, the 15 batches of medicinal material similarities greater than 0.9,6 batch of medicinal material similarity between 0.8-0.9.The similarity evaluation of analyzing each batch chromatographic peak specifically sees Table 1-2 and Fig. 1 b.
Table 1-221 criticizes erigeron breviscapus medicinal material sample principal character fingerprint similarity result of calculation
Embodiment 2: erigeron breviscapus bulk drug standard finger-print is set up
1, chromatographic condition and system suitability Agilent 1100 Series high performance liquid chromatographs; Zorbax SB-C18 (4.6mm * 250mm, 5 μ m) chromatographic column; Take acetonitrile-methyl alcohol-formic acid as mobile phase; Detect wavelength: 270nm, 330nm; Theoretical cam curve should be not less than 6000 by scutellarin peak calculating.
2, it is appropriate that the scutellarin reference substance is got in the preparation of object of reference solution, adds the solution that methyl alcohol is made 0.4mg/mL, and get final product.
3, the preparation precision of need testing solution takes the methanol solution that appropriate test sample is mixed with 1mg/mL, and get final product.
4, accurate object of reference solution and each 5 μ L of need testing solution of drawing of determination method, the injection liquid chromatography, measure and get final product respectively.
5, erigeron breviscapus bulk drug standard finger-print is set up
(1) according to 20 test datas of 10 batch sample, form the common pattern of erigeron breviscapus bulk drug finger-print.
(2) have 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in erigeron breviscapus bulk drug standard finger-print, be respectively, No. 9 peaks (S): scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
(3) each principal character fingerprint peaks relative retention time, relative peak area and shared total peak area ratio see Table 2-1.With object of reference corresponding peak be S peak (No. 9 peaks), No. 11 the unimodal area percentage in peak does not reach baseline separation greater than 20%, 11, No. 12 peak, joint account should group the total peak area at peak, its difference must not be greater than ± 20%; All the other each characteristic fingerprint peak area number percents are not done requirement all less than 10% to its difference.Characteristic fingerprint peak area number percent sum must not be less than 95%, and non-characteristic fingerprint peak area number percent sum is less than 5%.
Table 2-1 erigeron breviscapus bulk drug finger-print examination criteria
The standard finger-print of erigeron breviscapus bulk drug is as shown in Fig. 2 a.
" similarity evaluation " that different batches erigeron breviscapus bulk drug finger-print and erigeron breviscapus bulk drug standard finger-print common pattern adopt Chinese Pharmacopoeia Commission to recommend (2004B) calculates, and similarity answers 〉=0.9.
6, Methodological Analysis
(1) stability
1. similarity
Get the need testing solution of same lot number sample, respectively 0,1.5,3.0,4.5,9.0,10.5h measures, the overall picture of finger-print of observing directly perceived is without significant change, calculate with similarity, the finger-print that records on instrument on the same stage and the similarity of its common pattern collection of illustrative plates are between 0.999-1.000, RSD is 0.045%, shows that the composition of need testing solution is stable in 10.5h.The results are shown in Table 2-2 and Fig. 2 b.
Table 2-2 erigeron breviscapus bulk drug fingerprint similarity computational stability test findings
2. relative retention time, relative peak area and shared total peak area ratio
Get the need testing solution of same lot number sample, respectively 0,1.5,3.0,4.5,9.0,10.5h measures, wherein unimodal area accounts for total peak area more than or equal to 9,11,12,13, No. 15 characteristic fingerprint peaks of 5%, its relative retention time, relative peak area and shared total peak area ratio, RSD shows that all less than 3% the composition of need testing solution is stable in 10.5h.The results are shown in Table 2-3.
Table 2-3 erigeron breviscapus bulk drug finger-print principal character fingerprint peaks relative retention time stability test result
(2) precision
1. similarity
Get the need testing solution of same lot number sample, continuous 6 sample introductions are measured, the finger-print that observation post directly perceived records, and overall picture is without significant change; Calculate with similarity, the finger-print that records on instrument on the same stage and the similarity of its common pattern collection of illustrative plates are that 1.000, RSD is 0.00%, show that precision is up to specification.The results are shown in Table 2-4.
Table 2-4 erigeron breviscapus bulk drug fingerprint similarity calculates Precision test result
2. relative retention time, relative peak area and shared total peak area ratio
Get the need testing solution of same lot number sample, continuous 6 sample introductions are measured, wherein unimodal area accounts for total peak area more than or equal to 9,11,12,13, No. 15 characteristic fingerprint peaks of 5%, its relative retention time, relative peak area and shared total peak area ratio, RSD shows that all less than 3% precision is up to specification.The results are shown in Table 2-5.
Table 2-5 erigeron breviscapus bulk drug Fingerprints fingerprint peaks relative retention time Precision test result
7,10 batches of erigeron breviscapus bulk drug sample finger-prints
Get 10 batches of erigeron breviscapus bulk drug samples, make respectively need testing solution, the every batch of need testing solution replicate determination 2 times, the finger-print that observation post directly perceived records, overall picture is without significant change; Calculate with similarity, the finger-print and the similarity of its common pattern collection of illustrative plates that record on instrument on the same stage are with median method calculating, between 0.994-0.999; Calculate with the average method, between 0.995-0.999, show that similarity is up to specification.The results are shown in Table 2-6.
Table 2-610 criticizes erigeron breviscapus bulk drug sample fingerprint similarity result of calculation
Embodiment 3: the foundation of erigeron breviscapus standard preparation finger-print
1, chromatographic condition and system suitability Agilent 1100 Series high performance liquid chromatographs; Zorbax SB-C18 (4.6mm * 250mm, 5 μ m) chromatographic column; Take acetonitrile-methyl alcohol-formic acid as mobile phase; Detect wavelength: 270nm, 330nm; Theoretical cam curve should be not less than 6000 by scutellarin peak calculating.
2, it is appropriate that the scutellarin reference substance is got in the preparation of object of reference solution, adds the solution that methyl alcohol is made 0.4mg/mL, and get final product.
3, the preparation precision of need testing solution takes the methanol solution that appropriate test sample is mixed with 0.02g/mL, and get final product.
4, determination method accurate object of reference solution and each 5 μ L of need testing solution of drawing respectively, inject high performance liquid chromatograph, measures and get final product.
5, erigeron breviscapus standard preparation finger-print is set up
(1) according to 20 test datas of 10 batch sample, form the finger-print common pattern of erigeron breviscapus preparation.
(2) have 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in erigeron breviscapus standard preparation finger-print, be respectively: No. 9 peaks (S): scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
(3) each characteristic fingerprint peak relative retention time, relative peak area and shared total peak area ratio see Table 3-1.With object of reference corresponding peak be S peak (No. 9 peaks), No. 11 the unimodal area percentage in peak does not reach baseline separation greater than 20%, 11, No. 12 peak, joint account should group the total peak area at peak, its difference must not be greater than ± 20%; All the other each total peak area number percents are not done requirement all less than 10% to its difference.Characteristic fingerprint peak area number percent sum must not be less than 95%, and non-characteristic fingerprint peak area number percent sum is less than 5%.
Table 3-1 erigeron breviscapus preparation finger examination criteria
Erigeron breviscapus standard preparation finger-print is as shown in Fig. 3 a.
Different batches erigeron breviscapus preparation finger and erigeron breviscapus standard preparation finger-print common pattern adopt " similarity evaluation " of Chinese Pharmacopoeia Commission (2004B) to calculate, and similarity answers 〉=0.9.
6, Methodological Analysis
(1) stability
1. similarity
Get the need testing solution of same lot number sample, respectively 0,1.5,3.0,4.5,7.5,9.0h measures, the overall picture of finger-print of observing directly perceived is without significant change, calculate with similarity, the finger-print that records on instrument on the same stage and the similarity of its common pattern collection of illustrative plates are between 0.999-1.000, RSD is 0.045%, shows that the composition of need testing solution is stable in 9.0h.The results are shown in Table 3-2 and Fig. 3 b.
Table 3-2 erigeron breviscapus preparation finger similarity computational stability test findings
2. relative retention time, relative peak area and shared total peak area ratio
Get the need testing solution of same lot number sample, respectively 0,1.5,3.0,4.5,7.5,9.0h measures, wherein unimodal area accounts for total peak area more than or equal to 9,11,12,13, No. 15 characteristic fingerprint peaks of 5%, the RSD of its relative retention time, relative peak area and shared total peak area ratio shows that all less than 3% the composition of need testing solution is stable in 9.0h.The results are shown in Table 3-3.
Table 3-3 erigeron breviscapus preparation finger principal character fingerprint peaks relative retention time stability test result
(2) precision
1. similarity
Get the need testing solution of same lot number sample, continuous 6 sample introductions are measured, the finger-print that observation post directly perceived records, and overall picture is without significant change; Calculate with similarity, the chromatographic fingerprinting that records on instrument on the same stage and the similarity of its gained common pattern collection of illustrative plates are that 1.000, RSD is 0.00%, show that precision is up to specification.Detailed results sees Table 3-4.
Table 3-4 erigeron breviscapus preparation finger similarity is calculated Precision test result
2. relative retention time, relative peak area reach total peak ratio
Get the need testing solution of same lot number sample, continuous 6 sample introductions are measured, wherein unimodal area accounts for total peak area more than or equal to 9,11,12,13, No. 15 characteristic fingerprint peaks of 5%, the RSD of its relative retention time, relative peak area and shared total peak area ratio shows that all less than 3% precision is up to specification.Detailed results sees Table 3-5.
Table 3-5 erigeron breviscapus preparation finger principal character fingerprint peaks relative retention time Precision test result
7,10 batches of erigeron breviscapus formulation samples finger-prints
Get 10 batches of erigeron breviscapus formulation samples, make respectively need testing solution, the every batch of need testing solution replicate determination 2 times, the finger-print that observation post directly perceived records, overall picture is without significant change; Calculate with similarity, the chromatographic fingerprinting and the similarity of its common pattern collection of illustrative plates that record on instrument on the same stage are with median method calculating, between 0.989-0.999; Calculate with the average method, between 0.993-0.997, show that similarity is up to specification, the results are shown in Table 3-6.
Table 3-610 criticizes erigeron breviscapus formulation samples fingerprint similarity result of calculation
Embodiment 4: the standard finger-print evaluation
1, the figure spectrum correlation is estimated
Described erigeron breviscapus medicinal material, erigeron breviscapus bulk drug and standard preparation finger-print are compared, and the concrete condition of analysis is as follows:
(1) removed impurity in the process from the medicinal material to the bulk drug, chromatographic peak in bulk drug and standard preparation finger-print is than greatly reducing in medicinal material standard finger-print, 1-8 peak in the medicinal material standard finger-print is all removed, kept 9-15 active component chromatographic peak, and each peak-to-peak ratio is also basically identical, 16-21 peak clearance has guaranteed bulk drug and the quality of the pharmaceutical preparations and pharmacologically active greater than 90.0%;
(2) from the medicinal material to the bulk drug again to preparation, new chromatographic peak does not all appear in finger-print, show that the chromatographic peak in bulk drug and preparation all derives from medicinal material, extract and production run is not introduced other impurity;
(3) standard finger-print of bulk drug and preparation does not have obvious difference, embodies stronger consistance;
(4) in bulk drug and preparation the shared ratio (accounting for total peak area number percent) of non-characteristic fingerprint peak sum all less than 5%;
(5) from the medicinal material to the bulk drug again to preparation, its total peak shows consistance preferably, thereby has clear and definite sibship and high correlativity.
The application of finger-print on erigeron breviscapus of 2, setting up
(1) described three clear demonstrations of finger-print and pointed out (totally 21 characteristic fingerprint peaks, 20 characteristic fingerprint peaks of erigeron breviscapus medicinal material, wherein 1 the unknown), 11 characteristic fingerprint peaks of 11 characteristic fingerprint peaks of erigeron breviscapus bulk drug and erigeron breviscapus preparation, and peak area and the ratio of each fingerprint peaks, the ratio of erigeron breviscapus medicinal material, erigeron breviscapus bulk drug and preparation basic substance and each basic substance is confirmed;
(2) described erigeron breviscapus medicinal material standard finger-print can be used as erigeron breviscapus evaluation of medical materials' quality standard and the foundation of the screening that feeds intake." similarity evaluation " that different batches medicinal materials fingerprint and erigeron breviscapus medicinal material standard finger-print adopt Chinese Pharmacopoeia Commission to recommend (2004B) calculates, and similarity 〉=0.9 side is qualified, and can feed intake;
(3) described erigeron breviscapus bulk drug and standard preparation finger-print can be used as the criteria of quality evaluation of erigeron breviscapus bulk drug and preparation." similarity evaluation " that different batches bulk drug and preparation finger and respective standard finger-print adopt Chinese Pharmacopoeia Commission to recommend (2004B) calculates, and similarity 〉=0.9 side is qualified;
(4) described erigeron breviscapus bulk drug and standard preparation finger-print show, when the extraction of erigeron breviscapus bulk drug and preparation are produced, medicinal material 1-8 impurity peaks is all removed, 9-15 active component peak all keeps, 16-21 peak clearance is greater than 90.0%, and between described two standard finger-prints without significant difference.Can be used as erigeron breviscapus bulk drug and the quality of the pharmaceutical preparations and control foundation, guarantee its stability, homogeneity.
(5) described erigeron breviscapus shows to the standard preparation finger-print from the medicinal material to the bulk drug again, the total fingerprint peaks of bulk drug and preparation all derives from medicinal material, and new chromatographic peak does not appear, show sibship clear and definite between the three and high correlativity, and show extract and production run in do not introduce other impurity.Described three standard finger-prints consist of a complete quality evaluation system, realization from the erigeron breviscapus medicinal material to the erigeron breviscapus bulk drug again to technique, the quality whole process supervision of preparation, be convenient to the quality of production technology is made an appraisal, and follow-up process modification and the perfect standard indication that provides are provided.
Claims (11)
1. the method for building up of the standard finger-print of an erigeron breviscapus medicinal material and erigeron breviscapus bulk drug and preparation, comprise the following steps: get scutellarin and be dissolved in right amount in methyl alcohol as object of reference solution; Get respectively again erigeron breviscapus medicinal material, erigeron breviscapus bulk drug and preparation and make need testing solution; Drawing respectively at last object of reference solution and test sample solution gets with the high performance liquid chromatograph test;
Wherein, in described high performance liquid chromatography test, the chromatographic column specification is 4.6mm * 250mm, 5 μ m; Mobile phase is acetonitrile-methyl alcohol-formic acid; Adopt the DAD detecting device, detect in wavelength 270nm and 330nm;
contain 21 principal character fingerprint peakses in the standard finger-print of described erigeron breviscapus medicinal material, the peak is arranged No. 1: the pyromeconic acid glucoside, No. 2 peaks: pyromeconic acid, No. 3 peaks: protocatechuic acid, No. 4 peaks: chlorogenic acid+P-hydroxybenzoic acid, No. 5 peaks: the unknown, No. 6 peaks: caffeic acid+multiradiate fleabane glycosides, No. 7 peaks: syringic acid, No. 8 peaks: fleabane flower glycosides, No. 9 peaks: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 16 peaks: 6-hydroxyl Kaempferol, No. 17 peaks: high baicalein, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
2. the method for building up of standard finger-print according to claim 1, it is characterized in that, after described erigeron breviscapus medicinal material extracts with methyl alcohol, use dissolve with methanol solution, filter, obtain erigeron breviscapus medicinal material need testing solution, then obtain the standard finger-print of erigeron breviscapus medicinal material with the high performance liquid chromatograph test.
3. the method for building up of standard finger-print according to claim 2, is characterized in that, described methanol solution for dissolving is that volume ratio is 60% methanol solution.
4. the method for building up of the described standard finger-print of according to claim 1 to 3 any one, it is characterized in that, described erigeron breviscapus bulk drug is dissolved in methyl alcohol, obtain erigeron breviscapus bulk drug need testing solution, then obtain the standard finger-print of erigeron breviscapus bulk drug with the high performance liquid chromatograph test.
5. the method for building up of standard finger-print according to claim 4, it is characterized in that, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus bulk drug, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
6. the method for building up of standard finger-print according to claim 5, it is characterized in that, in the standard finger-print of described erigeron breviscapus bulk drug, the number percent of the shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 39.39%, No. 10 peaks: 0.95%, No. 11 peaks: 32.39%, No. 12 peaks: 5.90%, No. 13 peaks: 6.48%, No. 14 peaks: 1.51%, No. 15 peaks: 8.16%, No. 18 peaks: 0.55%, No. 19 peaks: 0.33%, No. 20 peaks: 1.30% and No. 21 peak: 0.42%.
7. the method for building up of the described standard finger-print of according to claim 1 to 3 any one, it is characterized in that, described erigeron breviscapus preparation is dissolved in methyl alcohol, obtains erigeron breviscapus preparation need testing solution, then obtains the standard finger-print of erigeron breviscapus preparation with the high performance liquid chromatograph test.
8. the method for building up of standard finger-print according to claim 7, it is characterized in that, contain 11 principal character fingerprint peakses total with the standard finger-print of described erigeron breviscapus medicinal material in the standard finger-print of described erigeron breviscapus preparation, described 11 characteristic fingerprint peaks all derive from the erigeron breviscapus medicinal material, and identical with 11 characteristic fingerprint peaks in the erigeron breviscapus bulk drug, comprise the peak No. 9: scutellarin, No. 10 peaks: 3, the 4-DCQ, No. 11 peaks: 3, the 5-DCQ, No. 12 peaks: 1, the 5-DCQ, No. 13 peaks: Erigeroster B, No. 14 peaks: oil lamp first element, No. 15 peaks: 4, the 5-DCQ, No. 18 peaks: 1, 3, 5-three caffeoyl quinic acids, No. 19 peaks: 3, 4, 5-three caffeoyl quinic acids, No. 20 peaks: apiolin, No. 21 peaks: Erigeroster B ethyl ester.
9. the method for building up of standard finger-print according to claim 8, it is characterized in that, in the standard finger-print of described erigeron breviscapus bulk drug, the number percent of the shared total peak area of each principal character fingerprint peaks is respectively, No. 9 peaks: 37.10%, No. 10 peaks: 1.68%, No. 11 peaks: 26.75%, No. 12 peaks: 8.20%, No. 13 peaks: 6.44%, No. 14 peaks: 1.31%, No. 15 peaks: 8.65%, No. 18 peaks: 1.26%, No. 19 peaks: 0.83%, No. 20 peaks: 3.15% and No. 21 peak: 0.58%.
10. the application of the standard finger-print that makes of the described method of according to claim 1 to 9 any one in erigeron breviscapus medicinal material and erigeron breviscapus bulk drug and preparation detect.
11. application according to claim 10, it is characterized in that, described application comprises that being applied to the erigeron breviscapus medicinal material feeds intake and optimize screening, be applied to erigeron breviscapus bulk drug and preparation basic substance and ratio-dependent, be applied to the erigeron breviscapus bulk drug and preparation process instructs, is applied to the erigeron breviscapus bulk drug and the quality of the pharmaceutical preparations is controlled.
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| CN104655748B (en) * | 2015-01-28 | 2017-09-19 | 云南植物药业有限公司 | A kind of erigeron breviscapus granule finger-print and its method for building up and application |
| CN105628835B (en) * | 2015-09-11 | 2018-02-09 | 贵州医科大学 | The multicomponent content assaying method of Sheepear Inula Herb medicinal material |
| CN107782835B (en) * | 2017-12-08 | 2019-10-29 | 云南农业大学 | A kind of method of efficient liquid phase detection fleabane flower ingredient |
| CN110346494B (en) * | 2019-06-03 | 2021-09-28 | 云南农业大学 | Method for simultaneously measuring contents of 4 effective components in erigeron breviscapus |
| CN110988234B (en) * | 2019-10-24 | 2022-06-07 | 昆明龙津药业股份有限公司 | Quality control method of erigeron breviscapus medicinal material |
| CN112067708B (en) * | 2020-06-23 | 2021-06-01 | 云南生物谷药业股份有限公司 | One-measurement-multiple-evaluation quantity detection method for erigeron breviscapus injection |
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