CN106018594B - A kind of construction method of qi-regulating and heart-soothing tablet HPLC characteristic spectrums - Google Patents
A kind of construction method of qi-regulating and heart-soothing tablet HPLC characteristic spectrums Download PDFInfo
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- CN106018594B CN106018594B CN201610326033.4A CN201610326033A CN106018594B CN 106018594 B CN106018594 B CN 106018594B CN 201610326033 A CN201610326033 A CN 201610326033A CN 106018594 B CN106018594 B CN 106018594B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
The construction method of qi-regulating and heart-soothing tablet characteristic spectrum, includes the following steps：(1) preparation of reference solution：It takes aurantiin accurately weighed, adds methanol that every solution, the preparation of (2) test solution is made：Take qi-regulating and heart-soothing tablet finely ground, accurately weighed, precision plus methanol supersound process supply less loss methanol, filter；(3) chromatographic condition is set；(4) it measures：Precision draws reference solution and test solution injects high performance liquid chromatograph, obtains qi-regulating and heart-soothing tablet characteristic spectrum.The method established of the present invention, can than more comprehensively reflecting the type and quantity of contained chemical composition, can Efficient Characterization its quality, be conducive to control product quality comprehensively, overcome the one-sidedness of protoplasm amount control method, be effective supplement of quality control matter method.Chinese medicine characteristic spectrum technology is a kind of quality control model of multi objective, can comprehensively, synthetically reflect and control traditional Chinese medicine quality.
Description
Technical field
The invention belongs to field of medicaments, are related to a kind of quality determining method of Chinese medicine.
Background technology
Chinese patent drug qi-regulating and heart-soothing tablet is by Radix Angelicae Sinensis, agalloch eaglewood, Poria cocos, root of Aucklandia lappa Decne, rhizoma cyperi (vinegar system), turmeric, curcuma zedoary (vinegar system), Pu
Huang, fingered citron, excrementum pteropi, dried orange peel, the dried immature fruit of citron orange (stir-fry), green peel (vinegar system), Fructus Aurantii (stir-fry), malt (stir-fry), citron, trigone (vinegar system), pellet
The 18 taste Chinese herbal medicines such as ginseng composition.This product major function is solution liver depression, and promoting the circulation of qi is stagnant, the obstruction of qi in the chest of dispelling.For qi depression to blood stasis disease coronary heart disease, the heart
Angina, cardiac arrhythmia, shortness of breath abdominal distension, chest distress and palpitation symptoms.Performed standard is recorded in Chinese medicine ministry standard, standard number at present：
WS3-B-0827-91, differentiates without thin layer in standard and the quality controls items such as assay, is unfavorable for the quality of effective monitoring drug
And curative effect.To improve qi-regulating and heart-soothing tablet drug quality, method of quality control research is carried out to full side early period, but because Chinese medicine is multiple
Square preparation flavour of a drug are more, and complicated component interferes with each other the features such as being not easy effectively to detect, and only several taste of traditional Chinese medicine can be set up exclusive in side
The strong method for qualitative and quantitative detection of property, is unable to overall monitor product quality, does not find the quality in relation to qi-regulating and heart-soothing tablet also at present
Research and standard upgrading data of literatures.
Chinese medicine compound prescription is the principal mode of tcm clinical practice medication, and quality is the key that clinical efficacy.How science is established
Reasonably, embodying the method for quality control of Chinese medicine compound prescription feature is just particularly important, and Chinese medicine compound prescription method of quality control one
It is directly one of Chinese medicine scientific research field hot research direction.How the feature efficiency index of scientific and reasonable selected quality control at
Point, effective quality control is carried out to Chinese medicine compound prescription, is the key point for establishing Chinese medicine compound prescription method of quality control.Therefore, originally
We have carried out characteristic spectrum experimental study to qi-regulating and heart-soothing tablet in invention, establish characteristic spectrum assay method, and to establishing
Method has carried out the methods of repeatability, precision and has learned verification test, the results showed that method is stablized feasible.Current standard is unfavorable for having
The quality and curative effect of effect control drug, do not find quality research and standard upgrading pertinent literature in relation to qi-regulating and heart-soothing tablet also at present
Data.
Invention content
An object of the present invention is to provide the construction method of qi-regulating and heart-soothing tablet HPLC characteristic spectrums, includes the following steps：
(1) preparation of reference solution：Take aurantiin reference substance appropriate, it is accurately weighed, add methanol that every lml is made and contains 100
The solution of~300 μ g to get.
(2) preparation of test solution：It takes qi-regulating and heart-soothing tablet finely ground, takes about 1~3g, it is accurately weighed, precision addition 60~
80% 12.5~50ml of methanol, weighed weight are ultrasonically treated 10~30 minutes, let cool, less loss is supplied with 60~80% methanol
Weight, filtration, take subsequent filtrate to get；
(3) chromatographic condition：Chromatographic column is using octadecylsilane chemically bonded silica as filler；Mobile phase is -0.1% phosphorus of acetonitrile
The gradient mixed solution of acid solution 3 → 70: 97 → 30；Ultraviolet detection wavelength is 286nm；Flow velocity is 1.0ml/min；
(4) it measures：Precision draws reference solution and each 10 μ l of test solution, high performance liquid chromatograph is injected, according to height
Effect liquid phase chromatogram method measures, and obtains qi-regulating and heart-soothing tablet characteristic spectrum.
(5) methodological study
Precision test：Qi-regulating and heart-soothing tablet is taken, test solution is made by step (2) method, continuous sample introduction 5 times, in accordance with the law
Detection.As a result, 5 relative retention time RSD values for measuring each shared peak in characteristic spectrum are 0~0.11%, show instrument essence
Density is good.
Stability test：Qi-regulating and heart-soothing tablet is taken, test solution is made by step (2) method, it is small 0,4,8,12 respectively
Shi Yifa is detected.As a result：4 times the relative retention time RSD values for measuring each shared peak in characteristic spectrum are 0~0.62%,
Show that test solution is stablized between 12 hours.
Repetitive test：It takes with a collection of qi-regulating and heart-soothing tablet, prepares 6 parts of test solutions by step (2) method, survey in accordance with the law
It is fixed.As a result, 6 relative retention time RSD values for measuring each shared peak in characteristic spectrum are 0~0.51%, show the weight of method
Renaturation is preferable.
Preferably, a concentration of of step (1) described reference solution contains 200 μ g reference substances per 1ml methanol.
Preferably, the preparation process of step (2) described test solution is as follows：It takes qi-regulating and heart-soothing tablet finely ground, takes about 2g, essence
Close weighed, 70% methanol 25ml is added in precision, and weighed weight is ultrasonically treated 20 minutes, lets cool, less loss is supplied with 70% methanol
Weight, filtration, take subsequent filtrate to get；
Preferably, step (3) described mobile phase is that the gradient mixing of -0.1% phosphoric acid solution 3 → 70: 97 → 30 of acetonitrile is molten
Liquid.
In a preferred embodiment, qi-regulating and heart-soothing tablet mobile phase gradient is as shown in table 1：
1 qi-regulating and heart-soothing tablet characteristic spectrum eluent gradient of table
| Time (minute) | Acetonitrile (%) | 0.1% phosphoric acid solution (%) |
| 0~35 | 3→25 | 97→75 |
| 35~40 | 25→30 | 75→70 |
| 40~55 | 30→70 | 70→30 |
| 55~70 | 70 | 30 |
| 70~75 | 70→3 | 30→97 |
It is a further object to provide the construction methods of the HPLC compare feature collection of illustrative plates of qi-regulating and heart-soothing tablet, according to preceding
It states method and builds 10 batches of qi-regulating and heart-soothing tablet HPLC characteristic spectrums, given birth to using chromatographic fingerprints of Chinese materia medica similarity evaluation system software
At the qi-regulating and heart-soothing tablet HPLC compare feature collection of illustrative plates being made of 10 shared peaks, it is aurantiin peak to share No. 5 peak in peak, the 6th
Number peak is aurantiamarin peak, and No. 7 peak is neohesperidin peak, and No. 8 peak is tanshin polyphenolic acid B peak.
It is with reference to the peak peaks S, when calculating opposite reservation of each characteristic peak with the peaks S with aurantiin peak in compare feature collection of illustrative plates
Between, within ± the 5% of specified value, the specified value is respectively the relative retention time：The peaks 0.22- 1, the peaks 0.28- 2,
The peaks 0.66- 3, the peaks 0.96- 4, the peaks 1.00- 5, the peaks 1.03- 6, the peaks 1.07- 7, the peaks 1.20- 8, the peaks 1.58- 9, the peaks 1.64- 10.
Qi-regulating and heart-soothing tablet sample is taken, is operated by above-mentioned same method, qi-regulating and heart-soothing tablet sample characteristic collection of illustrative plates is obtained, using Chinese medicine color
Spectrum finger-print similarity evaluation system software levies collection of illustrative plates to the reference material of qi-regulating and heart-soothing tablet and sample finger-print is analyzed,
It is qualified products that similarity, which is more than 0.90,.
Beneficial effects of the present invention are as follows：
(1) quality determining method of the invention can effectively monitor the quality of qi-regulating and heart-soothing tablet, ensure its quality stabilization,
Controllably.With method simplicity, precision is high, high repeatability and other advantages, can characteristic chemical constituent in Efficient Characterization qi-regulating and heart-soothing tablet, have
Conducive to the quality of monitoring product.
(2) method of the invention is using aurantiin, aurantiamarin, neohesperidin as reference substance, the qi-regulating and heart-soothing tablet HPLC of foundation
Compare feature collection of illustrative plates focuses on the tandem and correlation of each characteristic peak, focuses on whole facial feature and is conducive to overall monitor
The quality of product provides reliable foundation for the quality testing of drug.
(3) best UV absorption wavelength of the invention is 286nm.Using method in the present invention with DAD detectors to qi-regulating
Comfortable is measured, and the Detection wavelength result for occurring absorption maximum according to all chromatographic peaks in 3-D scanning figure is transferred respectively respectively
Chromatogram under wavelength, when Detection wavelength is 286, characteristic peak number is more, is detached between each peak area is higher and each characteristic peak
Effect is preferably also.
The method established of the present invention, can be than more comprehensively reflecting the type and quantity of contained chemical composition, can effective table
Its quality is levied, is conducive to control product quality comprehensively, overcomes the one-sidedness of protoplasm amount control method, is quality control matter method
Effectively supplement.Chinese medicine characteristic spectrum technology is a kind of quality control model of multi objective, comprehensively, synthetically can reflect and control
Traditional Chinese medicine quality is worthy to be popularized and applies.
Description of the drawings
Fig. 1 is qi-regulating and heart-soothing tablet HPLC characteristic spectrums；
Fig. 2 is 10 batch primitive character collection of illustrative plates of qi-regulating and heart-soothing tablet；
Fig. 3 is the compare feature collection of illustrative plates of qi-regulating and heart-soothing tablet.
Specific implementation mode
Embodiment 1 builds the HPLC characteristic spectrums of qi-regulating and heart-soothing tablet
1. instrument, reagent
1.1 instrument：1200 high performance liquid chromatograph of Agilent；Wear the double ternary high performance liquid chromatographs of peace；MS205DU types point
Analyse balance (Shanghai plum Teller-support benefit Instrument Ltd.)；Chromatographic column：Agilent ZORBAX SB-C18 (5 μm, 4.6 ×
250mm)。
1.2 reagent：Aurantiin reference substance, aurantiamarin reference substance, neohesperidin reference substance, tanshin polyphenolic acid B reference substance (are purchased from state
Food and medicine examines and determine research institute).Methanol is that analysis is pure, and acetonitrile, phosphoric acid are chromatographically pure, and water is purified water.
2. method
The preparation of 2.1 reference solutions：Take aurantiin reference substance appropriate, it is accurately weighed, add methanol that every lml is made and contains 200 μ
The solution of g to get.
The preparation of 2.2 test solutions：It takes qi-regulating and heart-soothing tablet finely ground, takes about 2g, accurately weighed, 70% first is added in precision
Alcohol 25ml, weighed weight are ultrasonically treated 20 minutes, let cool, the weight of less loss is supplied with 70% methanol, filter, take subsequent filtrate, i.e.,
；
2.3 chromatographic condition：Chromatographic column is using octadecylsilane chemically bonded silica as filler；Mobile phase is -0.1% phosphorus of acetonitrile
Acid solution is eluted by regulation gradient in table 1；Ultraviolet detection wavelength is 286nm；Flow velocity is 1.0ml/min；Number of theoretical plate is pressed
Aurantiin peak, which calculates, should be not less than 100000.
2.4 measuring：Precision draws reference substance solution and each 10 μ l of test solution, high performance liquid chromatograph is injected, according to height
Effect liquid phase chromatogram method measures, and obtains characteristic spectrum, as shown in Figure 1.
Embodiment 2 builds the HPLC compare feature collection of illustrative plates of qi-regulating and heart-soothing tablet
10 batch of qi-regulating and heart-soothing tablet (company A 8 batches, 1 batch, B companies, 1 batch, C companies) is taken, is detected by 1 condition of embodiment,
The HPLC collection of illustrative plates of 10 batch samples is obtained, as shown in Figure 2.By the comparison of 10 batches of HPLC collection of illustrative plates, using Chinese Pharmacopoeia, the committee pushes away
It recommends《Chromatographic fingerprints of Chinese materia medica similarity evaluation system》Software carries out similarity evaluation to HPLC collection of illustrative plates, determines common characteristic
Peak obtains qi-regulating and heart-soothing tablet compare feature collection of illustrative plates.
Contain 10 common characteristic peaks in qi-regulating and heart-soothing tablet compare feature collection of illustrative plates, wherein being aurantiin (No. 5 with reference to peak
Peak).
Similarity evaluation：With《Chromatographic fingerprints of Chinese materia medica similarity evaluation system》Software is analyzed, and test sample feature is calculated
The similarity of collection of illustrative plates and compare feature collection of illustrative plates, is as a result all higher than 0.90.Similarity evaluation the results are shown in Table 2.
Table 2HPLC characteristic spectrum similarity evaluation results
| Sample batch | Similarity | Sample batch | Similarity |
| 1 | 0.99 | 6 | 0.90 |
| 2 | 0.99 | 7 | 0.99 |
| 3 | 0.99 | 8 | 0.99 |
| 4 | 0.99 | 9 | 0.99 |
| 5 | 0.90 | 10 | 0.90 |
The qi-regulating and heart-soothing tablet HPLC compare feature collection of illustrative plates established by the above method is as shown in Figure 3.
The quality determining method of 3 qi-regulating and heart-soothing tablet of embodiment
Take 7 batches of qi-regulating and heart-soothing tablet samples (company A 3 batches, 1 batch, B companies, 1 batch, C companies, 1 batch, D companies, 1 batch, E companies) by real
The condition for applying example 1 is operated with method, qi-regulating and heart-soothing tablet sample characteristic collection of illustrative plates is obtained, using chromatographic fingerprints of Chinese materia medica similarity evaluation
System software analyzes qi-regulating and heart-soothing tablet compare feature collection of illustrative plates and sample characteristic collection of illustrative plates, and similarity is more than 0.90 for qualified production
Product.
Testing result shows that company A, B companies, C companies sample are qualified samples, and D companies, E companies sample are unqualified
Sample.
In conclusion the stability of quality determining method of the present invention is good, reproducible, and therefore, technical scheme of the present invention
With feasibility.Qualified qi-regulating and heart-soothing tablet is detected using quality determining method of the present invention, drug effect is definite, and effect is stablized.Below
It is illustrated using experimental example.
4 pharmacological experiment of embodiment
1. experiment material
1.1 experimental animal
250 scholar 20g, ICR mouse of rat, 20 scholar 2g, source：Jilin University's Bethune's laboratory animal center of medical college is qualified
Card number：SCXK (Ji) 2014-0003.
1.2 laboratory apparatus, reagent
BL-410 Biological Signal Collecting Systems：Purchased from Chengdu Tai Meng Co., Ltds；SOD, MDA, CK, LDH, AST kit
It is purchased from Nanjing and builds up biotechnology Co., Ltd.
1.3 experimental drug：
Experiment comparison medicine A used is that qualified qi-regulating and heart-soothing tablet is detected using quality determining method of the present invention, and comparison medicine E is
Underproof qi-regulating and heart-soothing tablet is detected using quality determining method of the present invention.
1.4 dose design
Qi-regulating and heart-soothing tablet clinical administration dosage be 17.622g (crude drug)/day/people, this experiment using clinic 2 multiple doses into
Row pharmacodynamic experiment compares, i.e., equivalent rat test dose is：3.2g (crude drug)/kg, mouse experiment dosage are:4.6g is (raw
Medicine)/kg.
2. experimental method
The effect of 2.1 pairs of Acute Myocardial Ischemia in Rats
Wistar male rats 50, are randomly divided into 5 groups, are sham-operation group respectively, model group, positive drug ginkgo leaf group,
Qi-regulating and heart-soothing tablet comparison medicine A, qi-regulating and heart-soothing tablet comparison medicine E.Daily gastric infusion 1 time, successive administration 7 days.Last time is administered
30min afterwards is injected intraperitoneally 3% yellow Jackets (1ml/kg) anesthetized animal, supine position is fixed on operating table, neck
Portion's unhairing, 75% alcohol disinfecting, the midsection skin about 0.5cm along suprasternal fossa, blunt separation push glandula submandibularis open, separation
Muscle before tracheae makes tracheae fully expose.In the 2nd~3 tracheae interannular promoting the circulation of qi pipe transection, incision length is no more than tracheae week
Tracheae content is cleared up in the 1/3 of diameter, is rapidly inserted into trachea cannula, and depth is 0.5~1cm.Connect toy lung ventilator, breathing
Frequency 90 times/min, 10~12ml of tidal volume, respiratory quotient 1:2.Left front chest unhairing, 75% alcohol disinfecting, left border of sternum are cut
Skin is about 2cm, successively blunt separation subcutaneous tissue, muscle, opens chest, drag hook chest expanding in the 4th~5 intercostal, passivity removes pericardium
Film, exposure heart, ramus descendens anterior arteriae coronariae sinistrae is ligatured under pulmonary conus and left auricle of heart, heart is put back to rapidly at 2~3mm
Thoracic cavity squeezes out intrathoracic gas, sutures the wall of the chest.Sham-operation group experimental implementation is essentially identical, only in the left front drop of coronary artery
Threading is without ligaturing at branch.Postoperative 6h, takes a blood sample through abdominal aorta, and 3000rmp/min centrifugations 10min prepares serum, for detecting
SOD, MDA, CK, LDH, AST.
After blood sampling, heart is taken out rapidly, with the physiological saline of precooling, is removed blood stains, is rejected the non-cardiac muscle such as extra fat
Tissue.- 20 DEG C keep overnight, and next day takes out from the apex of the heart to heart basal part is parallel and is cut into 5, is placed in 0.025%TTC solution,
It is incubated 15min in 37 DEG C of water-baths, this process will constantly shake dyeing liquor, be allowed to adequately contact cardiac muscular tissue.It is stood after taking-up
I.e. with normal saline flushing to extra dyestuff.It is analyzed by BI-2000 medical image analysis systems, calculates infarct face
Product.
Will be fixed in the neutral formalin solution for the heart 10% rinsed well, fixed through paraffin embedding, slice, dimethylbenzene
Dewaxing, gradient alcohol dehydration, HE dyeing, resinene mounting and etc..Observe the morphological change of rat heart muscle tissue.
As a result:
1. the influence pair rat blood serum SOD, MDA
SOD, MDA measurement result model group SOD vigor have reduction, MDA contents to have increase, relatively have with sham-operation group significantly
Difference (p<0.001)；Positive drug ginkgo leaf group SOD vigor has raising, MDA contents to be substantially reduced, and relatively has with model group significantly
Difference (p<0.001), qi-regulating and heart-soothing tablet comparison medicine A SOD contents are significantly increased, and MDA contents are substantially reduced, with model group ratio
Compared with there were significant differences (p<0.01), comparison medicine comparison medicine ESOD vigor has raising, (the p that compares that there were significant differences compared with model group<
0.05), MDA contents reduce, but there was no significant difference compared with model group.It is shown in Table 3.
Influence (x ± S) of the table 3 to rat blood serum SOD, MDA
The * * p compared with sham-operation group<0.01、***p<0.001,；The #p compared with model group<0.05, ##p<0.01, ###p<
0.001
2. the influence pair rat blood serum CK, LDH, AST
CK, LDH, AST measurement result model group CK, LDH, AST content obviously increase, and have compared with sham-operation group notable
Difference (p<0.001).Qi-regulating and heart-soothing tablet comparison medicine A CK, LDH, AST release increase, with difference (p compared with model group<
0.05、p<0.01)；Comparison medicine ECK releases have increase, with the more significant difference (p of model group<0.05), LDH, AST with
Model group compares that there was no significant difference.It is shown in Table 4.
Influence (x ± S) of the table 4 to rat blood serum CK, LDH, AST
The * * * p compared with sham-operation group<0.001；The #p compared with model group<0.05、##p<0.01
3. the influence pair rat myocardial infarction model area
Normal group of rat myocardial infarction model area estimation result has no myocardial ischemia, and model group myocardial ischemia is the most serious, reason
Comfortable comparison medicine A myocardial infarction area of gas is most light, is better than qi-regulating and heart-soothing tablet comparison medicine E, is shown in Table 5.
5 each group rat myocardial infarction model area percentage of table
The influence of 2.2 pairs of rat ventriculars
Wistar rats 40, half male and half female are randomly divided into 4 groups, are model group, positive drug ginkgo leaf group, qi-regulating respectively
Comfortable comparison medicine A, qi-regulating and heart-soothing tablet comparison medicine E.Daily gastric infusion 1 time, continuous 7 days, model control group gave isometric(al)
CMC.The 12h fasting before experiment of each group rat, can't help water.After last time administration 1h, 10% chloraldurate fiber crops are injected intraperitoneally
It is liquor-saturated.Rat lies on the back fixation, records II lead electrocardio with BL-410 Biological Signal Collecting Systems, stablizes 30s, discarding electrocardiogram not just
Normal rat.For aconitine (sterling) through Rat Tongue intravenously administrable, dosage is 20 μ g/kg weight, is injected in 3s.Observe rat electrocardio
There is the duration in time (incubation period) and arrhythmia cordis of arrhythmia cordis in the situation of change of figure, record rat.As a result：Qi-regulating
There is arrhythmic event after comfortable comparison medicine A, qi-regulating and heart-soothing tablet comparison medicine E injection aconitines and is longer than blank control group (p<
0.05、p<0.01、p<0.001)；Qi-regulating and heart-soothing tablet comparison medicine A restores normal ECG and restores normal ECG completely for the first time
Time is compared with model group, qi-regulating and heart-soothing tablet comparison medicine E early (P ＜ 0.05, p<0.01), there is significant difference, and present centainly
Timeliness imitates relationship, is shown in Table 6.
The 6 rat ventricular time of table compares (x ± S, n=10)
* P ＜ 0.05, * * P ＜ 0.01, * * * P ＜ 0.001 compared with the control group；The #P compared with qi-regulating and heart-soothing tablet comparison medicine E
＜ 0.05, ##P ＜ 0.01
The influence of 2.3 pairs of mouse oxygen deficit tolerances
ICR mouse, half male and half female, are randomly divided into 4 groups, blank group, positive drug Jenner are multigroup, qi-regulating and heart-soothing tablet pair by 40
According to medicine A, qi-regulating and heart-soothing tablet comparison medicine E, every group 10.Experimental group is administered daily 1 time, and continuous 7d, blank group gives isometric(al)
Purified water is respectively placed in mouse in the 250ml wide-mouth bottles demarcated through capacity after last dose 1h, a piece of filter paper built in bottle and
To absorb moisture content and carbon dioxide, bottle cap is coated with vaseline sealing, records the death time of mouse, that is, when surviving 10g soda limes
Between.
As a result, qi-regulating and heart-soothing tablet comparison medicine A can be obviously prolonged the mouse survival time, with the more significant difference of blank group
(p<0.05) and the mouse survival time is longer than qi-regulating and heart-soothing tablet comparison medicine E.It is shown in Table 7.
Influence (x ± s) of the table 7 to the mouse survival time
The * p compared with blank control group<0.05
2.4 conclusion
The resisting myocardial ischemia of qualified Chinese patent drug qi-regulating and heart-soothing tablet, anti-heart rate are monitored using quality determining method of the present invention not
It plays a role clearly together and curative effect is stablized.
Claims (5)
1. the construction method of qi-regulating and heart-soothing tablet characteristic spectrum, includes the following steps：
(1) preparation of reference solution：Take aurantiin reference substance appropriate, it is accurately weighed, add methanol that every lml is made and contains 100~300
The solution of μ g to get；
(2) preparation of test solution：It takes qi-regulating and heart-soothing tablet finely ground, takes 1~3g, accurately weighed, precision is added 60~80%
12.5~50ml of methanol, weighed weight are ultrasonically treated 10~30 minutes, let cool, the weight of less loss is supplied with 60~80% methanol,
Filtration, take subsequent filtrate to get；
(3) chromatographic condition：Chromatographic column is Agilent ZORBAX SB-C18 chromatographic columns；Mobile phase A is mutually acetonitrile, and B phases are
0.1% phosphoric acid solution；Mobile phase ratio is when gradient elution：0~35min, A phase：3~25%, B phases：97~75%；35~
40min, A phase：25~30%, B phases：75~70%；40~55min, A phase：30~70%, B phases：70~30%；55~
70min, A phase：70%, B phases：30%；70~75min, A phase：70~3%, B phases：30~97%；Ultraviolet detection wavelength is 240
~320nm；Flow velocity is 0.5~1.5ml/min；Column temperature is 25~45 DEG C；
(4) it measures：Precision draws reference solution and each 5~20 μ l of test solution, high performance liquid chromatograph is injected, according to efficient
Liquid chromatography for measuring obtains qi-regulating and heart-soothing tablet characteristic spectrum.
2. the construction method of qi-regulating and heart-soothing tablet characteristic spectrum according to claim 1, it is characterized in that：The test solution
Preparation process it is as follows：It takes qi-regulating and heart-soothing tablet finely ground, takes about 2g, accurately weighed, 70% methanol 25ml is added in precision, weighed heavy
Amount, be ultrasonically treated 20 minutes, let cool, the weight of less loss supplied with 70% methanol, filter, take subsequent filtrate to get.
3. the construction method of the HPLC compare feature collection of illustrative plates of qi-regulating and heart-soothing tablet, which is characterized in that according to any one of claim 1-2
The method builds 10 batches of qi-regulating and heart-soothing tablet HPLC characteristic spectrums, soft using chromatographic fingerprints of Chinese materia medica similarity evaluation system
Part generates the qi-regulating and heart-soothing tablet HPLC compare feature collection of illustrative plates being made of 10 shared peaks, and No. 5 peak is shaddock ped in the shared peak
Glycosides peak, No. 6 peak are aurantiamarin peak, and No. 7 peak is neohesperidin peak, and No. 8 peak is tanshin polyphenolic acid B peak.
4. according to the method described in claim 3, it is characterized in that, in the compare feature collection of illustrative plates, using aurantiin peak as reference
The peak peaks S, calculate the relative retention time of each characteristic peak and the peaks S, and the relative retention time is respectively：The peaks 0.22- 1, the peaks 0.28-
2, the peaks 0.66- 3, the peaks 0.96- 4, the peaks 1.00- 5, the peaks 1.03- 6, the peaks 1.07- 7, the peaks 1.20- 8, the peaks 1.58- 9, the peaks 1.64- 10.
5. the detection method of qi-regulating and heart-soothing tablet, which is characterized in that qi-regulating and heart-soothing tablet sample is taken, it is same by any one of claim 1-2
Method operates, and qi-regulating and heart-soothing tablet sample characteristic collection of illustrative plates is obtained, using chromatographic fingerprints of Chinese materia medica similarity evaluation system software to institute
It states sample characteristic collection of illustrative plates and claim 3-4 any one of them qi-regulating and heart-soothing tablet HPLC compare feature collection of illustrative plates is analyzed, phase
It is qualified products to be more than 0.90 like degree.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610326033.4A CN106018594B (en) | 2016-05-17 | 2016-05-17 | A kind of construction method of qi-regulating and heart-soothing tablet HPLC characteristic spectrums |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610326033.4A CN106018594B (en) | 2016-05-17 | 2016-05-17 | A kind of construction method of qi-regulating and heart-soothing tablet HPLC characteristic spectrums |
Publications (2)
| Publication Number | Publication Date |
|---|---|
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| CN106018594B true CN106018594B (en) | 2018-08-07 |
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Effective date of registration: 20220719 Address after: No. 39, Modified Road, Tonghua City, Jilin Province, 134000 Patentee after: Tongyao Pharmaceutical Group Corp. Address before: 130103 No. 1369 Shunda Road, Changchun High-tech Zone, Jilin Province Patentee before: JILIN XIUZHENG PHARMACEUTICAL NEW MEDICINE DEVELOPMENT Co.,Ltd. |