CN103760254A - Method for determining fingerprint spectrum of traditional Chinese medicine for treating coronary heart disease - Google Patents
Method for determining fingerprint spectrum of traditional Chinese medicine for treating coronary heart disease Download PDFInfo
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Abstract
The invention relates to a method for determining fingerprint spectrum of traditional Chinese medicine for treating coronary heart disease. The method comprises the following steps: preparing a test solution, preparing a reference solution, and carrying out gradient elution under the chromatographic conditions that an Agilent ZORBAX Extend C18 chromatographic column is used as a filling agent, a mobile phase A is methanol and a mobile phase B is 0.4% phosphoric acid water solution; determining under conditions that the flow velocity is at 1.0ml/min, the column temperature is at 35 DEG C and the detection wavelength is 230-240nm, thus obtaining the fingerprint spectrum of traditional Chinese medicine. The method has the characteristics of being simple and convenient, good in reproducibility, more in characteristic peaks, accurate and reliable, and the like, and the quality of the traditional Chinese medicine preparation can be controlled effectively by the established fingerprint spectrum.
Description
Technical field
The present invention relates to a kind of finger print measuring method for the treatment of the Chinese medicine preparation of coronary heart disease, is a kind ofly with high performance liquid chromatography fingerprint spectrum method, to measure the active principle composition in Chinese medicine preparation specifically, belongs to medical technical field.
Background technology
Chinese medicine is of long standing and well established in the history of China, the material carrier of preventing and curing diseases as the traditional Chinese medical science, and the inherent quality of itself is directly connected to clinical efficacy and safety.For this reason, the Chinese medicine preparation quality control system that foundation can be accepted by international pharmaceuticals industry is one of key issue of Development of Chinese Medicine.It is the important content of the modernization of Chinese medicine that traditional Chinese medicine quality is controlled, and fingerprint pattern technology is considered to the effective means that traditional Chinese medicine quality is controlled research.Traditional Chinese medicine fingerprint can reflect kind and the quantity of chemical substance in Chinese medicine more all sidedly, and Chinese medicine preparation carried out to the checking of quality controllability and product stability.In finger-print, a plurality of target components especially effective constituent are carried out to accurate quantitative analysis, traditional Chinese medicine quality is controlled more accurate, can more truly reflect that the quality condition of Chinese medicine and product form, and make the quality of product and stability more reliable.
Arklemin open capsule is to be produced without competition by Shaanxi step-length pharmaceutical Co. Ltd, and the national drug standards of carrying out are now numbered WS
3-155 (Z-025)-2005(Z), it has outstanding curative effect aspect treatment coronary heart disease, said preparation is to take Mongolian medicine's theory as a kind of new drug according to developing, by 5 taste medicines such as fructus choerospondiatis, the red sage root, cloves, borneol, Tabasheers, be prepared from, there is function promoting blood circulation and removing blood stasis, clearing and activating the channels and collaterals, promoting qi circulation and relieving pain, the treatment of the diseases such as coronary heart diseases and angina pectoris that cause for the obstruction of qi in the chest stagnation of the heart blood.For our first to file of this Chinese medicine preparation the patent of publication number CN1686294A, it discloses prescription and the preparation method of the Chinese medicine preparation of its treatment coronary heart disease, because having good clinical effectiveness and market sale, Chinese medicine preparation of the present invention has a extensive future, and at present to only tanshin polyphenolic acid B and tanshinone IIA content being controlled in the quality standard of said preparation, this is obviously far from being enough to controlling Chinese medicine preparation inherent quality.In recent years, fingerprint pattern technology has become the important means of Chinese medicine compound prescription quality control and evaluation, therefore technical solutions according to the invention, will carry out finger-print research to arklemin open capsule, for further improving the quality standard of this product, provide reference frame.
Summary of the invention
The object of the invention is to provide a kind of can control the fingerprint atlas detection method of arklemin open capsule quality comprehensively, effectively, reliably, by the method, obtain the finger-print information than more comprehensive Multiple components, thereby Chinese medicine preparation inherent quality is controlled reference frame is provided comprehensively for this reason.It is easy, accurate that the method has, and the finger-print of acquisition has the features such as characteristic peak is many, favorable reproducibility.
Chinese medicine preparation described in content of the present invention, all refer to the arklemin open capsule of being produced by Shaanxi step-length pharmaceutical Co. Ltd, it is to be prepared from by fructus choerospondiatis, the red sage root, cloves, borneol, Tabasheer 5 taste medicinal materials, and its concrete prescription proportioning consists of: fructus choerospondiatis 480g, red sage root 240g, cloves 60g, borneol 30g, Tabasheer 30g;
The concrete preparation method of said preparation is as follows: gets fructus choerospondiatis 120g and is ground into fine powder, be divided into two parts, and standby, residue fructus choerospondiatis is ground into meal, according to the percolation (appendix IO of Chinese Pharmacopoeia version in 2005) under liquid extract and extract item, with 70% ethanol, makes solvent, carry out diacolation, collect percolate, reclaim ethanol, and be concentrated into relative density 1.30～1.35(50 ℃), add a fructus choerospondiatis fine powder, mix thoroughly, 60 ℃ of drying under reduced pressure, are ground into fine powder.Get the red sage root and extract three times, use for the first time alcohol heating reflux 1.5 hours, filter, filtrate recycling ethanol, and to be concentrated into relative density be 1.30(55～60 ℃); Use for the second time 50% alcohol heating reflux 1.5 hours, filter; Boiling for the third time 2 hours, filter, merge filtrate second, third time, reclaim ethanol, and to be concentrated into relative density be 1.40(55～60 ℃), merge with primary concentrate, mix, being concentrated into relative density is 1.35～1.39(55～60 ℃), add another part of fructus choerospondiatis fine powder, mix thoroughly, 60 ℃ of drying under reduced pressure, are ground into fine powder.Tabasheer is ground into fine powder, is divided into two parts, standby.Cloves is extracted volatile oil by steam distillation, and volatile oil is evenly sprayed in a Tabasheer fine powder, mixes, airtight; Borneol mixes with another part of Tabasheer fine powder, and porphyrize mixes with above-mentioned each fine powder, incapsulates, and makes 1000, obtains.
Be below the finger print measuring method that above-mentioned Chinese medicine preparation is set up, the method comprises following step:
(a) preparation of Chinese medicine preparation solution: take the about 1.0g of Chinese medicine preparation content, mix, accurately weighed, put in 100mL tool plug conical flask, add 10mL70% methyl alcohol, weigh, ultrasonic 30min, let cool, the weight of supplying less loss with 70% methyl alcohol, shakes up, and 0.45 μ m miillpore filter filters, get subsequent filtrate, obtain;
(b) chromatographic condition: be filling agent with octadecylsilane chemically bonded silica; Mobile phase is methyl alcohol: 0.4% phosphate aqueous solution, and its proportioning is methyl alcohol 3%～100%, 0.4% phosphate aqueous solution 0～97%, carries out linear gradient elution;
Detect wavelength: 230～240nm; Column temperature: 35 ℃, flow velocity: 1.0mLmin
-1, sample size: 5 μ L.
Further, the chromatographic condition in described step (b), the condition of its preferred mobile phase linear gradient elution is:
During 0～15min, the shared volume ratio of methyl alcohol is that the volume ratio that 3 → 28%, 0.4% phosphate aqueous solution accounts for is 97% → 72%;
During 15～30min, the shared volume ratio of methyl alcohol is that the volume ratio that 28 → 30%, 0.4% phosphate aqueous solution accounts for is 72% → 70%;
During 30～35min, the shared volume ratio of methyl alcohol is that the volume ratio that 30 → 32%, 0.4% phosphate aqueous solution accounts for is 70 → 68%;
During 35～47min, the shared volume ratio of methyl alcohol is that the volume ratio that 32%, 0.4% phosphate aqueous solution accounts for is 68%;
During 47～56min, the shared volume ratio of methyl alcohol is that the volume ratio that 32 → 41%, 0.4% phosphate aqueous solution accounts for is 68 → 59%;
During 56～70min, the shared volume ratio of methyl alcohol is that the volume ratio that 41 → 48%, 0.4% phosphate aqueous solution accounts for is 59 → 52%;
During 70～100min, the shared volume ratio of methyl alcohol is that the volume ratio that 48 → 88%, 0.4% phosphate aqueous solution accounts for is 52 → 12%;
During 100～105min, the shared volume ratio of methyl alcohol is that the volume ratio that 88 → 100%, 0.4% phosphate aqueous solution accounts for is 12 → 0%;
Detection wavelength is: 0-55min, 230nm; 55-62min, 240nm; 62-105min, 230nm.
Further, according to above-mentioned set up finger print measuring method, measure the finger-print that Chinese medicine preparation of the present invention obtains, through compare of analysis, obtained 30 total peaks, and take tanshin polyphenolic acid B absorption peak as reference, the relative retention time obtaining is: No. 1 peak: 0.067 ± 1.198%; No. 2 peaks: 0.075 ± 1.070%; No. 3 peaks: 0.086 ± 0.695%; No. 4 peaks: 0.103 ± 0.588%; No. 5 peaks: 0.136 ± 0.670%; No. 6 peaks: 0.158 ± 0.507%; No. 7 peaks: 0.166 ± 0.372%; No. 8 peaks: 0.209 ± 0.346%; No. 9 peaks: 0.283 ± 0.541%; No. 10 peaks: 0.301 ± 0.201%; No. 11 peaks: 0.317 ± 0.231%; No. 12 peaks: 0.462 ± 0.443%; No. 13 peaks: 0.549 ± 0.629%; No. 14 peaks: 0.623 ± 0.684%; No. 15 peaks: 0.746 ± 0.520%; No. 16 peaks: 0.842 ± 0.690%; No. 17 peaks: 0.884 ± 0.589%; No. 18 peaks: 1.000; No. 19 peaks: 1.077 ± 0.112%; No. 20 peaks: 1.110 ± 0.128%; No. 21 peaks: 1.121 ± 0.162%; No. 22 peaks: 1.237 ± 0.196%; No. 23 peaks: 1.558 ± 0.366%; No. 24 peaks: 1.587 ± 0.309%; No. 25 peaks: 1.624 ± 0.450%; No. 26 peaks: 1.642 ± 0.366%; No. 27 peaks: 1.676 ± 0.358%; No. 28 peaks: 1.712 ± 0.364%; No. 29 peaks: 1.746 ± 0.398%; No. 30 peaks: 1.837 ± 0.360%.
Further, the main total peak that utilizes reference substance and the present invention to set up in finger-print carries out Analysis deterrmination: No. 4 peaks are gallic acid, and No. 6 peaks are Sodium Danshensu, and No. 7 peaks are protocatechuic acid, No. 8 peaks are protocatechualdehyde, No. 9 peaks are caffeic acid, and No. 14 peaks are rutin, and No. 16 peaks are Rosmarinic acid, No. 18 peaks are tanshin polyphenolic acid B, No. 22 peaks are eugenol, and No. 27 peaks are Cryptotanshinone, and No. 29 peaks are tanshinone IIA.
In the characteristic peak of the finger-print generating in the invention described above Chinese medicine preparation finger-print, select each total peak area relatively to account for total peak that has peak area ratio example >=3%, and to take tanshin polyphenolic acid B absorption peak in reference fingerprint be reference, the relative peak area specifically obtaining is: No. 6 peaks: 0.266; No. 8 peaks: 0.160; No. 22 peaks: 0.266; No. 27 peaks: 1.744; No. 29 peaks: 0.226.
The finger print measuring method that the present invention sets up, has that reappearance is good, the reliable feature of stability, the true and false that can be used as Chinese medicine preparation differentiate and composition detection in application, specifically have following beneficial effect:
1, the finger-print of setting up by method provided by the invention, can effectively characterize 30 characteristic components in treatment Chinese medicine preparation of the present invention and form, and 11 compositions are differentiated, is conducive to embody its inherent quality.
2, investigated the stability of Chinese medicine preparation solution of the present invention, result shows, Chinese medicine preparation solution of the present invention, in 24h, keeps stable, and the precision of the relative retention time of measuring and peak area and replica test result good (RSD is 0.01%～4.9%); Through the fingerprint similarity of 10 batches of Chinese medicine preparations of the present invention is calculated, find that its similarity is between 0.90～0.999.The Chinese medicine preparation quality control method of the treatment coronary heart disease of setting up is described, easy and simple to handle, method is reliable, can this type of Chinese medicine preparation quality be carried out science, effectively be evaluated, and confirm to apply to quality control in actual production, meet the demand of enterprise to production quality control.
3, technical solution of the present invention is selected to take tanshin polyphenolic acid B as reference, is because content of danshinolic acid B is higher in Chinese medicine preparation of the present invention, and the retention time in the finger-print obtaining and peak area more stable, reappearance is good.Modern pharmacology experiment shows, tanshin polyphenolic acid B can suppress platelet aggregation, and can obviously improve the nervous function defect of cerebral reperfusion injury rat, it has antithrombotic effect, this and Chinese medicine preparation of the present invention to have function promoting blood circulation and removing blood stasis similar, also can carry out quantitative and qualitative analysis detection to active principle composition in Chinese medicine preparation.The fingerprint pattern quality determination method that the present invention sets up, for enterprise, improving quality standard provides reference frame.
Accompanying drawing explanation
Fig. 1 is Chinese medicine preparation contrast characteristic fingerprint pattern of the present invention.
Fig. 2-Figure 12 is respectively the finger-print of reference substance gallic acid, Sodium Danshensu, protocatechuic acid, protocatechualdehyde, caffeic acid, rutin, Rosmarinic acid, tanshin polyphenolic acid B, eugenol, Cryptotanshinone, tanshinone IIA reference substance solution.
Figure 13 is the finger-print of 10 batches of Chinese medicine preparations of the present invention.
Embodiment
In order more to fully understand enforcement of the present invention, below by typical embodiment, the present invention is described further.
Unless otherwise defined, the technical term using in patent application specification of the present invention and claims or scientific terminology should be has the ordinary meaning that the personage of general technical ability understands under the present invention in field." Chinese medicine preparation " described in patent application specification of the present invention and claims all refers to arklemin open capsule, and its prescription proportioning and preparation method partly have definite implication at above summary of the invention.
Embodiment 1: the foundation of Chinese medicine preparation finger print measuring method of the present invention
1.1 instruments and reagent
Agilent1260 (quaternary pump G1311B; Automatic sampler G1329B; Column oven G1316A; Diode array detector G4212B; LAB open Data Processing in Chromatography Workstation); Electronic analytical balance (model BP211D, d=0.01mg, German Sai Duolisi balance company limited); Ultrasonic cleaner (model: KQ-250DE type, Kunshan Ultrasonic Instruments Co., Ltd.); Agilent ZORBAX Extend-C
18chromatographic column (250mm * 4.6mm, 5 μ m).
1.2 reagent
10 batches of Chinese medicine preparations of the present invention are produced by Shaanxi step-length pharmaceutical Co. Ltd, and 10 batch sample numberings are respectively Y
1, Y
2, Y
3, Y
4, Y
5, Y
6, Y
7, Y
8, Y
9, Y
10, the accurate word Z20020055 of traditional Chinese medicines, is capsule.Methyl alcohol is chromatographically pure (U.S. Fisher reagent company), and water is Wahaha board pure water, and it is pure that other reagent are analysis.
The preparation of 1.3 need testing solutions
Get the about 1g of Chinese medicine preparation content, mix, accurately weighed, put in 100mL tool plug conical flask, add 10mL70% methyl alcohol, weigh, ultrasonic (100W, 45kHz) 30min, lets cool, and supplies the weight of less loss with 70% methyl alcohol, shakes up 0.45
μ m miillpore filter filters, and gets subsequent filtrate and get final product.
The preparation of 1.4 reference substance solution
Get gallic acid, Sodium Danshensu, protocatechuic acid, protocatechualdehyde, caffeic acid, rutin, Rosmarinic acid, tanshin polyphenolic acid B, eugenol, Cryptotanshinone, tanshinone IIA reference substance appropriate, accurately weighed, add methyl alcohol and make every 1ml respectively containing the mixed solution of gallic acid 20 μ g, Sodium Danshensu 10 μ g, protocatechuic acid 20 μ g, protocatechualdehyde 10 μ g, caffeic acid 10 μ g, rutin 60 μ g, Rosmarinic acid 100 μ g, tanshin polyphenolic acid B 150 μ g, eugenol 70 μ g, Cryptotanshinone 20 μ g, tanshinone IIA 10 μ g, obtain.
1.5 chromatographic conditions:
Chromatographic column: Agilent ZORBAX Extend-C18 chromatographic column (250mm * 4.6mm, 5 μ m); Column temperature: 35 ℃, flow velocity: 1.0mLmin
-1, take methyl alcohol as mobile phase A, take 0.4% phosphate aqueous solution as Mobile phase B, the program of according to the form below 1 volumetric concentration preparation ratio, carries out gradient elution.
Table 1 eluent gradient elution requirement
Detect wavelength: 0-55min, 230nm; 55-62min, 240nm; 62-105min, 230nm.
Accurate need testing solution and the reference substance solution 5ul injection liquid chromatography drawn measured: record 105min chromatogram, obtain the finger-print of Chinese medicine preparation of the present invention and reference substance solution under above-mentioned " 1.5 chromatographic condition ".
1.6 determining of total peak
Get the sample of the Chinese medicine preparation of 10 different batches, by " 1.3 " below legal system, obtain solution, press " 1.5 lower condition chromatograms " each sample introduction 1 time, record chromatogram, data importing < < similarity evaluation A version > > is carried out to difference appraisal and global similarity evaluation to chromatographic peak, investigate the consistance of chromatographic peak similarity.Using 30 total peaks of common pattern as reference, calculate relative retention time and the relative peak area at total peak.
Experimental result shows: in test sample characteristic spectrum, present 30 characteristic peaks, wherein 11 peaks should be respectively with corresponding identical with reference to peak retention time; With corresponding peak, tanshin polyphenolic acid B object of reference peak be S peak, the retention time of tanshin polyphenolic acid B is 57.916min, peak area is 5982.998.Calculate the time that relatively remains with of its characteristic peak, its relative retention time should be setting ± 5% within.
Concrete each peak relative retention time of Chinese medicine preparation of the present invention is: No. 1 peak: 0.067 ± 1.198%; No. 2 peaks: 0.075 ± 1.070%; No. 3 peaks: 0.086 ± 0.695%; No. 4 peaks: 0.103 ± 0.588%; No. 5 peaks: 0.136 ± 0.670%; No. 6 peaks: 0.158 ± 0.507%; No. 7 peaks: 0.166 ± 0.372%; No. 8 peaks: 0.209 ± 0.346%; No. 9 peaks: 0.283 ± 0.541%; No. 10 peaks: 0.301 ± 0.201%; No. 11 peaks: 0.317 ± 0.231%; No. 12 peaks: 0.462 ± 0.443%; No. 13 peaks: 0.549 ± 0.629%; No. 14 peaks: 0.623 ± 0.684%; No. 15 peaks: 0.746 ± 0.520%; No. 16 peaks: 0.842 ± 0.690%; No. 17 peaks: 0.884 ± 0.589%; No. 18 peaks: 1.000; No. 19 peaks: 1.077 ± 0.112%; No. 20 peaks: 1.110 ± 0.128%; No. 21 peaks: 1.121 ± 0.162%; No. 22 peaks: 1.237 ± 0.196%; No. 23 peaks: 1.558 ± 0.366%; No. 24 peaks: 1.587 ± 0.309%; No. 25 peaks: 1.624 ± 0.450%; No. 26 peaks: 1.642 ± 0.366%; No. 27 peaks: 1.676 ± 0.358%; No. 28 peaks: 1.712 ± 0.364%; No. 29 peaks: 1.746 ± 0.398%; No. 30 peaks: 1.837 ± 0.360%.
In Chinese medicine preparation finger-print of the present invention, in 30 total peaks of feature, through compare of analysis, determined that No. 4 peaks are gallic acid, No. 6 peaks are Sodium Danshensu, No. 7 peaks are protocatechuic acid, and No. 8 peaks are protocatechualdehyde, and No. 9 peaks are caffeic acid, No. 14 peaks are rutin, No. 16 peaks are Rosmarinic acid, and 18 peaks are tanshin polyphenolic acid B, and No. 22 peaks are eugenol, No. 27 peaks are Cryptotanshinone, and No. 29 peaks are tanshinone IIA.
In the characteristic peak of the finger-print generating in Chinese medicine preparation finger, select each total peak area relatively to account for total peak that has peak area ratio example >=3%, and to take tanshin polyphenolic acid B absorption peak in reference fingerprint be reference, the relative peak area specifically obtaining is: No. 6 peaks: 0.266; No. 8 peaks: 0.160; No. 22 peaks: 1.744; No. 27 peaks: 0.224; No. 29 peaks: 0.226.
1.6 methodological studies:
1.6.1 precision test
Get Chinese medicine preparation sample Y of the present invention
5, by " 1.3 " below legal system, obtain solution, by " 1.5 " lower condition, repeat sample introduction 6 times, record chromatographic peak retention time and peak area.Calculate the similarity of these 6 characteristic spectrums all higher than 0.999, the results are shown in Table 2; Take peak 18(tanshin polyphenolic acid B) as with reference to peak.The RSD of the relative retention time of each total chromatographic peak is between 0.0458%～0.4879%; The RSD of relative peak area is between 0.0194%～4.8488%.Meet mensuration requirement, show that instrument precision is good.
Table 2 Chinese medicine preparation HPLC characteristic spectrum precision test similarity analysis result
1.6.2 replica test
Get 6 parts of the Chinese medicine preparations of the present invention of same lot number, by " 1.3 " below legal system, obtain solution, by " 1.5 " lower each sample introduction of condition 1 time, record chromatogram, take peak 18(tanshin polyphenolic acid B) as with reference to peak.Calculate the similarity of these 6 characteristic spectrums all higher than 0.999, the results are shown in Table 3; The RSD of the relative retention time of each total chromatographic peak is between 0.0117%～0.1351%; The RSD of relative peak area is between 0.1753%～4.8824%.Meet mensuration requirement, show that the repeatability of method is good.
Table 3 Chinese medicine preparation HPLC characteristic spectrum replica test similarity analysis result
1.6.3 stability test
Get and be numbered Y
5sample, by " 1.3 " below legal system, obtain solution, by " 1.5 " lower condition, respectively at 0h, 2h, 4h, 8h, 12h, 24h, record chromatographic peak area, the similarity that calculates these 6 characteristic spectrums is 0.999, the results are shown in Table 4; The RSD of the relative retention time of each total chromatographic peak is between 0.0168%～0.1542%; The RSD of relative peak area is between 0.0702%～2.2485%.Meet mensuration requirement, show that need testing solution is good at 24h internal stability.
Table 4 Chinese medicine preparation HPLC characteristic spectrum stability test similarity analysis result
The establishment of embodiment 2 Chinese medicine preparation finger-print of the present invention
The preparation of 2.1 need testing solutions
Get the about 1g of Chinese medicine preparation content of 10 different lot numbers, accurately weighed, put in 100mL tool plug conical flask, add 10mL70% methyl alcohol, weigh, ultrasonic (100W, 45kHz) 30min, lets cool, with 70% methyl alcohol, supply the weight of less loss, shake up, 0.45 μ m miillpore filter filters, and gets subsequent filtrate and get final product.
2.2 chromatographic conditions:
Chromatographic column: Agilent ZORBAX Extend-C18 chromatographic column (250mm * 4.6mm, 5 μ m); Column temperature: 35 ℃, flow velocity: 1.0mLmin
-1, take methyl alcohol as mobile phase A, take 0.4% phosphate aqueous solution as Mobile phase B, the program of according to the form below 5 volumetric concentration preparation ratios, carries out gradient elution.
Table 5 eluent gradient elution requirement
Detect wavelength: 0-55min, 230nm; 55-62min, 240nm; 62-105min, 230nm.
The accurate need testing solution of drawing 10 different batches, difference injection liquid chromatography, adopt above-mentioned " 2.2 chromatographic condition " to record 105min chromatogram, import < < similarity evaluation A version > >, each collection of illustrative plates superposes and carries out Auto-matching, time window width is 0.5min, 30 total chromatographic peaks have been demarcated altogether, median method generates contrast feature spectrogram, as benchmark, calculate 10 batch sample collection of illustrative plates and the similarity that contrasts collection of illustrative plates, stability between evaluating batch.Result shows, the similarity of 10 batch samples is good, all, more than 0.903, the results are shown in Table 6.
Table 610 batch Chinese medicine preparation similarity is investigated
Above experimental result shows, the method stability is reliable, can be used as the method for the quality of controlling Chinese medicine preparation of the present invention, and can be used as and differentiate that the present invention treats the discrimination method of the true and false of coronary heart disease Chinese medicine preparation.
Finally it should be noted that: the present invention is not limited to above-mentioned specific embodiments, above-mentioned specific embodiments is only schematic, guiding, rather than restrictive.Those of ordinary skill in the art, under the enlightenment of this instructions, in every case in spirit of the present invention and essential scope, any change of doing, is equal to and replaces and improve, all within protection scope of the present invention.
Claims (5)
1. a finger print measuring method for the treatment of the Chinese medicine preparation of coronary heart disease, described Chinese medicine preparation is to be prepared from by 480 parts of fructus choerospondiatis, 240 parts of the reds sage root, 60 parts of cloves, 30 parts of borneols, 30 parts of medicinal materials of Tabasheer, it is characterized in that, the method comprises following step:
(a) preparation of need testing solution: take Chinese medicine preparation content 1.0g, mix, accurately weighed, put in tool plug conical flask, add 70% methyl alcohol 10mL, weigh, ultrasonic 30min, lets cool, and supplies the weight of less loss with 70% methyl alcohol, shake up, filter, get subsequent filtrate, obtain;
(b) chromatographic condition: be filling agent with octadecylsilane chemically bonded silica; Mobile phase is methyl alcohol: 0.4% phosphate aqueous solution, and its volume proportion is that methyl alcohol accounts for 3%～100%, 0.4% phosphate aqueous solution accounts for 0～97%, carries out linear gradient elution; Detect wavelength: 230～240nm; Column temperature: 35 ℃, flow velocity: 1.0mLmin
-1, sample size: 5 μ L.
2. finger print measuring method as claimed in claim 1, is characterized in that, in the chromatographic condition of described step (b), the linear gradient elution condition that its mobile phase adopts is:
During 0～15min, the shared volume ratio of methyl alcohol is that the volume ratio that 3 → 28%, 0.4% phosphate aqueous solution accounts for is 97% → 72%;
During 15～30min, the shared volume ratio of methyl alcohol is that the volume ratio that 28 → 30%, 0.4% phosphate aqueous solution accounts for is 72% → 70%;
During 30～35min, the shared volume ratio of methyl alcohol is that the volume ratio that 30 → 32%, 0.4% phosphate aqueous solution accounts for is 70 → 68%;
During 35～47min, the shared volume ratio of methyl alcohol is that the volume ratio that 32%, 0.4% phosphate aqueous solution accounts for is 68%;
During 47～56min, the shared volume ratio of methyl alcohol is that the volume ratio that 32 → 41%, 0.4% phosphate aqueous solution accounts for is 68 → 59%;
During 56～70min, the shared volume ratio of methyl alcohol is that the volume ratio that 41 → 48%, 0.4% phosphate aqueous solution accounts for is 59 → 52%;
During 70～100min, the shared volume ratio of methyl alcohol is that the volume ratio that 48 → 88%, 0.4% phosphate aqueous solution accounts for is 52 → 12%;
During 100～105min, the shared volume ratio of methyl alcohol is that the volume ratio that 88 → 100%, 0.4% phosphate aqueous solution accounts for is 12 → 0%;
Detection wavelength is: 0-55min, 230nm; 55-62min, 240nm; 62-105min, 230nm.
3. finger print measuring method as claimed in claim 1 or 2, is characterized in that, has 30 characteristic peaks, and take tanshin polyphenolic acid B absorption peak as reference according to the method in the finger-print generating, and the relative retention time obtaining is: No. 1 peak: 0.067 ± 1.198%; No. 2 peaks: 0.075 ± 1.070%; No. 3 peaks: 0.086 ± 0.695%; No. 4 peaks: 0.103 ± 0.588%; No. 5 peaks: 0.136 ± 0.670%; No. 6 peaks: 0.158 ± 0.507%; No. 7 peaks: 0.166 ± 0.372%; No. 8 peaks: 0.209 ± 0.346%; No. 9 peaks: 0.283 ± 0.541%; No. 10 peaks: 0.301 ± 0.201%; No. 11 peaks: 0.317 ± 0.231%; No. 12 peaks: 0.462 ± 0.443%; No. 13 peaks: 0.549 ± 0.629%; No. 14 peaks: 0.623 ± 0.684%; No. 15 peaks: 0.746 ± 0.520%; No. 16 peaks: 0.842 ± 0.690%; No. 17 peaks: 0.884 ± 0.589%; No. 18 peaks: 1.000; No. 19 peaks: 1.077 ± 0.112%; No. 20 peaks: 1.110 ± 0.128%; No. 21 peaks: 1.121 ± 0.162%; No. 22 peaks: 1.237 ± 0.196%; No. 23 peaks: 1.558 ± 0.366%; No. 24 peaks: 1.587 ± 0.309%; No. 25 peaks: 1.624 ± 0.450%; No. 26 peaks: 1.642 ± 0.366%; No. 27 peaks: 1.676 ± 0.358%; No. 28 peaks: 1.712 ± 0.364%; No. 29 peaks: 1.746 ± 0.398%; No. 30 peaks: 1.837 ± 0.360%.
4. finger print measuring method as claimed in claim 3, it is characterized in that: No. 4 peaks are gallic acid, No. 6 peaks are Sodium Danshensu, and No. 7 peaks are protocatechuic acid, No. 8 peaks are protocatechualdehyde, No. 9 peaks are caffeic acid, and No. 14 peaks are rutin, and No. 16 peaks are Rosmarinic acid, No. 18 peaks are tanshin polyphenolic acid B, No. 22 peaks are eugenol, and No. 27 peaks are Cryptotanshinone, and No. 29 peaks are tanshinone IIA.
As claim 1 or 2 finger print measuring methods the true and false of this Chinese medicine preparation differentiate and composition detection in application.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103940929A (en) * | 2014-05-05 | 2014-07-23 | 山东丹红制药有限公司 | Method for detecting traditional Chinese medicine injection for treating cardiovascular and cerebrovascular diseases |
| CN104777253A (en) * | 2015-04-17 | 2015-07-15 | 国药控股深圳中药有限公司 | Establishment method for Guanxindanshen capsule fingerprint spectrum and fingerprint spectrum thereof |
| CN105287605A (en) * | 2015-12-01 | 2016-02-03 | 陕西步长制药有限公司 | Drug composition |
| CN105738502A (en) * | 2016-01-31 | 2016-07-06 | 张春辉 | Characteristic chromatogram establishment method of refined coronary tablets and application |
| CN106950289A (en) * | 2016-01-07 | 2017-07-14 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN109298091A (en) * | 2018-10-24 | 2019-02-01 | 安徽安科余良卿药业有限公司 | The standard feature map construction method and its quality determining method of wind eggplant Pingchuan ointment |
| CN110940748A (en) * | 2019-12-09 | 2020-03-31 | 健民药业集团股份有限公司 | HPLC fingerprint detection method of child Jinqing granules |
| CN111366672A (en) * | 2020-04-24 | 2020-07-03 | 劲牌有限公司 | Detection method of health wine fingerprint |
| CN112946118A (en) * | 2021-02-01 | 2021-06-11 | 贵州金桥药业有限公司 | Method for measuring medicine fingerprint and fingerprint thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030152654A1 (en) * | 1998-03-19 | 2003-08-14 | Xie De Long | Ginkgo biloba composition, method to prepare the same and uses thereof |
| CN1814206A (en) * | 2005-12-08 | 2006-08-09 | 咸阳步长医药科技发展有限公司 | Chinese medicine capsule for treating coronary heart clisease and angina pectoris, and quality control method |
| CN101954000A (en) * | 2009-07-16 | 2011-01-26 | 陕西步长制药有限公司 | Traditional Chinese medicinal dispersible tablets treating coronary disease and angina pectoris, preparation method thereof and detection method thereof |
| CN102866211A (en) * | 2011-12-31 | 2013-01-09 | 山西振东泰盛制药有限公司 | Detection method of fingerprint spectrum of Guanxinning injection |
-
2014
- 2014-01-03 CN CN201410003476.0A patent/CN103760254B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030152654A1 (en) * | 1998-03-19 | 2003-08-14 | Xie De Long | Ginkgo biloba composition, method to prepare the same and uses thereof |
| CN1814206A (en) * | 2005-12-08 | 2006-08-09 | 咸阳步长医药科技发展有限公司 | Chinese medicine capsule for treating coronary heart clisease and angina pectoris, and quality control method |
| CN101954000A (en) * | 2009-07-16 | 2011-01-26 | 陕西步长制药有限公司 | Traditional Chinese medicinal dispersible tablets treating coronary disease and angina pectoris, preparation method thereof and detection method thereof |
| CN102866211A (en) * | 2011-12-31 | 2013-01-09 | 山西振东泰盛制药有限公司 | Detection method of fingerprint spectrum of Guanxinning injection |
Non-Patent Citations (2)
| Title |
|---|
| 刘刚等: "变换波长HPLC测定七味广枣丸中丁香酚、木香烃内酯和去氢木香内酯的含量", 《中国药学杂志》 * |
| 马久太等: "HPLC 法测定冠心舒通胶囊中没食子酸的含量", 《世界中医药》 * |
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| CN103940929A (en) * | 2014-05-05 | 2014-07-23 | 山东丹红制药有限公司 | Method for detecting traditional Chinese medicine injection for treating cardiovascular and cerebrovascular diseases |
| CN103940929B (en) * | 2014-05-05 | 2015-11-04 | 山东丹红制药有限公司 | A kind of detection method for the treatment of the traditional Chinese medicine injection of cardiovascular and cerebrovascular disease |
| CN104777253A (en) * | 2015-04-17 | 2015-07-15 | 国药控股深圳中药有限公司 | Establishment method for Guanxindanshen capsule fingerprint spectrum and fingerprint spectrum thereof |
| CN105287605B (en) * | 2015-12-01 | 2017-12-12 | 陕西步长制药有限公司 | A kind of pharmaceutical composition |
| CN105287605A (en) * | 2015-12-01 | 2016-02-03 | 陕西步长制药有限公司 | Drug composition |
| CN106950289A (en) * | 2016-01-07 | 2017-07-14 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN106950289B (en) * | 2016-01-07 | 2019-10-29 | 陕西步长制药有限公司 | A kind of coronary disease treatment capsule one is surveyed comments detection method of content more |
| CN105738502A (en) * | 2016-01-31 | 2016-07-06 | 张春辉 | Characteristic chromatogram establishment method of refined coronary tablets and application |
| CN109298091A (en) * | 2018-10-24 | 2019-02-01 | 安徽安科余良卿药业有限公司 | The standard feature map construction method and its quality determining method of wind eggplant Pingchuan ointment |
| CN110940748A (en) * | 2019-12-09 | 2020-03-31 | 健民药业集团股份有限公司 | HPLC fingerprint detection method of child Jinqing granules |
| CN110940748B (en) * | 2019-12-09 | 2022-03-15 | 健民药业集团股份有限公司 | HPLC fingerprint detection method of child Jinqing granules |
| CN111366672A (en) * | 2020-04-24 | 2020-07-03 | 劲牌有限公司 | Detection method of health wine fingerprint |
| CN112946118A (en) * | 2021-02-01 | 2021-06-11 | 贵州金桥药业有限公司 | Method for measuring medicine fingerprint and fingerprint thereof |
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