CN104849364A - Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof - Google Patents
Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof Download PDFInfo
- Publication number
- CN104849364A CN104849364A CN201510224392.4A CN201510224392A CN104849364A CN 104849364 A CN104849364 A CN 104849364A CN 201510224392 A CN201510224392 A CN 201510224392A CN 104849364 A CN104849364 A CN 104849364A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- finger
- oral liquid
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a canzhiling oral solution fingerprint map building method. The map includes the following steps of preparation of a test solution, preparation of a reference solution, testing through a high performance liquid chromatograph and processing of data and a map. The invention further discloses a Canzhiling oral solution fingerprint map and a method for utilizing the fingerprint map to control quality of a Canzhiling oral solution. The Canzhiling oral solution fingerprint map building method is simple in operation, stable, reliable, high in accuracy and high in separation degree, the fingerprint map is high in stability and reproducibility and large in information quantity, and the fingerprint map is adopted as a quality control means for the Canzhiling oral solution, so that one-sidedness caused by judging of overall quality of a preparation by testing one or two chemical ingredients is avoided, and probability of artificial processing in order to enabling quality to be up to standards is lowered; samples of multiple batches are analyzed systematically, so that quality of the Canzhiling oral solution can be evaluated more comprehensively and scientifically, and product quality and efficacy are guaranteed.
Description
Technical field
The present invention relates to traditional Chinese medicine fingerprint analytical approach, particularly join method for building up and the finger-print of ginseng branch tuckahoe oral liquid that obtains of method thus of branch tuckahoe oral liquid HPLC finger-print.
Background technology
Ginseng branch tuckahoe oral liquid is by Radix Codonopsis, cassia twig, the root of herbaceous peony, Radix Glycyrrhizae (honey is processed), Poria cocos, rhizoma zingiberis, polygala root (system, stir-fry), grass-leaved sweetflag, keel and the compound Chinese medicinal preparation of oyster ten kinds of medicinal materials through manufacturing meticulously, now prove that it contains various bioactivators, there is good beneficial temperature sun, the curative effect of calming the nerves of reducing phlegm, be used for the treatment of the deficiency of heart-QI card that light moderate stages alzheimer's disease manifests, the diseases such as main manifestations is forgetful, palpitaition, dizziness, insomnia, weak breath complaint, tongue nature is light, apathy, spiritlessness and weakness, feeble pulse are unable.In ginseng branch tuckahoe oral liquid Radix Codonopsis act as calm the nerves bright, bushing gas; Polygala root, calamus, Poria cocos etc. then play logical thinking, the heart-yang that shakes, change turbid phlegm.Alzheimer's disease patient memory, intelligence and viability and cognitive function effectively can be improved for clinical.Show through pharmaceutical research, ginseng branch tuckahoe oral liquid can improve all segment pathology changes of senile dementia very effectively, the Spatial memory ability of alzheimer model of a syndrome rat can be improved, improve the phenomenon that its Cortical neurons fiber thickens and increases, and the symptom that its hippocampus cones comes off can be treated.
Ginseng branch tuckahoe oral liquid is as a kind of compound Chinese medicinal preparation, and prescription medicinal material more (being made up of 10 taste medicinal materials), composition is very complicated various.In " ginseng branch tuckahoe oral liquid standard " (State Food and Drug Administration's standard), adopt liquid phase chromatography to establish the content assaying method of Paeoniflorin and cinnamic acid two compositions.But ginseng branch tuckahoe oral liquid is compound preparation, merely by detecting one, quantitatively to control its drug quality be obviously incomplete to two kind of index components, can not embody the Integral Characteristic of compound Chinese medicinal preparation.Therefore, exploitation is set up a kind of simple and easy to do, and accuracy is high, and repeatability is good and can reflect that multicomponent fingerprint atlas detection method contained by compound preparation is for the quality control of ginseng branch tuckahoe oral liquid with evaluate very necessary as much as possible.
Summary of the invention
For above-mentioned the deficiencies in the prior art, the object of this invention is to provide seed ginseng branch tuckahoe oral liquid fingerprint map construction method and a finger-print thereof.The present invention is by the research to ginseng branch tuckahoe oral liquid finger-print, provide a kind of easy and simple to handle, stable, precision is high and favorable reproducibility, can be used for joining the quality testing of branch tuckahoe oral liquid and the method for evaluation, compensate for the deficiency of existing method of quality control, make that the Quality Control Technology of ginseng branch tuckahoe oral liquid is more perfect, science.
For achieving the above object, the present invention adopts following technical proposals:
One seed ginseng branch tuckahoe oral liquid fingerprint map construction method, comprises the following steps:
(1) preparation of need testing solution: get ginseng branch tuckahoe oral liquid, add methyl alcohol, ultrasonic extraction, 0.5-1 hour is placed in refrigeration, returns to room temperature, filters, gets subsequent filtrate, obtain need testing solution;
(2) preparation of reference substance solution: get cinnamic acid, Paeoniflorin, liquiritin, albiflorin, the ammonium glycyrrhetate reference substance of drying under reduced pressure to constant weight, add Methanol respectively and become reference substance solution;
(3) measure: precision measures need testing solution and reference substance solution respectively, inject high performance liquid chromatograph and measure, record chromatogram;
(4) with finger-print software, gained collection of illustrative plates is processed, branch tuckahoe oral liquid finger-print must be joined;
In step (3), the liquid phase chromatogram condition of mensuration is: chromatographic column is phenomenex synergi Hydro-RP chromatographic column (4 μ, 250 × 4.6mm; Namely packing material size is 4 μm, and column length is 250mm, and column internal diameter is 4.6mm); Mobile phase A is 0.3% phosphate aqueous solution (volume fraction), and Mobile phase B is acetonitrile; Gradient elution; Flow rate of mobile phase is 0.8ml/min; Column temperature is 30 DEG C; Determined wavelength is 230nm.
In step (3), in gradient elution process, being changed to of mobile phase A and Mobile phase B: 0-5min, mobile phase A 95%-95%, Mobile phase B 5%-5%; 5-10min, mobile phase A 95%-92%, Mobile phase B 5%-8%; 10-20min, mobile phase A 92%-80%, Mobile phase B 8%-20%; 20-25min, mobile phase A 80%-77.5%, Mobile phase B 20%-22.5%; 25-40min, mobile phase A 77.5%-67%, Mobile phase B 22.5%-33%; 40-50min, mobile phase A 67%-60%, Mobile phase B 33%-40%; 50-60min, mobile phase A 60%-50%, Mobile phase B 40%-50%; 60-80min, mobile phase A 50%-15%, Mobile phase B 50%-85%; 80-90min, mobile phase A 15%-5%, Mobile phase B 85%-95%; 90-100min, mobile phase A 5%-95%, Mobile phase B 95%-5%.(being volumn concentration).
Concrete, in step (1), the preparation method of need testing solution is: precision measures 1.25ml and joins branch tuckahoe oral liquid in 25ml volumetric flask, add methanol constant volume to scale, ultrasonic extraction 5min, places 0.5 hour, returns to room temperature in 4 DEG C of refrigerators, through 0.22 μm of organic system membrane filtration, get subsequent filtrate and namely obtain need testing solution.
In step (2), the preparation method of reference substance solution is:
Precision takes cinnamic acid standard items 0.0103g, puts in 100mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain cinnamic acid solution, and concentration is 0.103mg/mL;
Precision takes Paeoniflorin standard items 0.0302g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain Paeoniflorin solution, and concentration is 3.02mg/mL;
Precision takes liquiritin standard items 0.0204g, puts in 25mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain liquiritin solution, and concentration is 0.816mg/mL;
Precision takes albiflorin standard items 0.0101g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain albiflorin solution, and concentration is 1.01mg/mL;
Precision takes ammonium glycyrrhetate standard items 0.0198g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain ammonium glycyrrhetate solution, and concentration is 1.98mg/mL.
In step (3), the accurate volume measuring need testing solution and reference substance solution is 10 μ L respectively.
It should be noted that, the construction method of finger-print of the present invention is through scientific experiment and screens and obtain, and be not that the routine of this area is selected, the present invention is mainly to the preparation method of need testing solution, chromatographic condition, determined wavelength and analysis time etc., condition to be carried out preferably.
Wherein: the 1) investigation of need testing solution compound method: test compares the impact of Extraction solvent extension rate on sample solution Detection results, as diluted 40 times, 20 times and 10 times, result dilutes 20 times Detection results using methyl alcohol as Extraction solvent is good.
2) selection of chromatographic condition and optimization: investigated different phosphate acid buffer proportioning (0.1%, the 0.2% and 0.3%) impact on chromatogram separating effect in mobile phase.Test findings shows: when in mobile phase A, phosphate buffer solution proportioning is 0.3%, chromatographic resolution effect is better, and baseline is steady, and peak shape is symmetrical, and degree of separation is better, and therefore mobile phase is defined as: A phase (0.3% phosphoric acid water)-B phase (acetonitrile).After adjustment mobile phase different time wash-out ratio, the retention time of each chromatographic peak is moderate, and baseline is steady, not easily drifts about, improves the degree of separation of chromatogram simultaneously, effectively prevent the conditions of streaking of chromatogram, be conducive to the determination and analysis of chromatographic fingerprinting.
3) selection of determined wavelength: the chromatogram under each wavelength adopting photodiode array detector (PDAD) to scan at 190 ~ 400nm compares analysis, and set 8 wavelength and detect simultaneously and (comprise 220nm, 230nm, 235nm, 237nm, 240nm, 245nm, 254nm and 290nm), testing result shows: not only baseline is steady for the chromatogram of 230nm place detection, and chromatographic peak is more, contains much information, the separating effect of each chromatographic peak is better, and peak area is also larger.Therefore, select 230nm as determined wavelength.Investigated the chromatogram under other wavelength according to the requirement of tentative standard, its result display ginseng branch tuckahoe oral liquid chromatographic fingerprinting at different wavelengths has good similarity simultaneously.
4) selection of analysis time: the chromatogram that have recorded 2h when selecting the elution time of finger-print.Result shows that 85min does not have obvious chromatographic peak to occur later substantially, simultaneously in order to look after the otherness of batch sample, ensures that the characteristic peak of all batch sample can both be detected, and therefore selects 100min as analysis time.
5) selection of column temperature: test the impact of four different column temperatures (as 25 DEG C, 30 DEG C, 35 DEG C and 40 DEG C) on finger-print testing result.When result display column temperature is 30 DEG C, chromatographic peak retention time is suitable for, and baseline is steady, and each chromatographic peak degree of separation is better, and peak shape is symmetrical, therefore selects column temperature to be 30 DEG C.
6) selection of flow velocity: test four flow velocitys (0.7ml/min, 0.8ml/min, 0.9ml/min and 1.0ml/min) to the impact of finger-print testing result.Result shows: when flow velocity is 0.8ml/min, and separating effect is best, and each chromatographic peak retention time is suitable for, and degree of separation is good, and baseline is steady, and peak shape is symmetrical, therefore selects flow velocity to be 0.8ml/min.
Join the ginseng branch tuckahoe oral liquid finger-print that branch tuckahoe oral liquid fingerprint map construction method obtains as previously mentioned, the total peak in this finger-print is 11.
Present invention also offers the method for a seed ginseng branch tuckahoe oral liquid quality control, comprise the following steps:
(1) structure of ginseng branch tuckahoe oral liquid finger-print to be measured: get ginseng branch tuckahoe oral liquid to be measured, build the finger-print of ginseng branch tuckahoe oral liquid to be measured as stated above;
(2) finger-print of ginseng branch tuckahoe oral liquid to be measured and reference sample finger-print are carried out similarity system design, to evaluate the quality of ginseng branch tuckahoe oral liquid to be measured, computing method are correlation coefficient process, similarity be not less than 0.900 oral liquid up-to-standard.If similarity is less than 0.900, show that batch sample mass discrepancy is larger.
In step (2), the construction method of described reference sample finger-print is: the similarity first calculating each sample and other sample, then select the sample maximum with other sample similarity sum to be reference sample, its finger-print is reference sample finger-print.
Beneficial effect of the present invention:
(1) the present invention adopts phosphate aqueous solution-acetonitrile system, and adopt the finger-print of the method establishment of gradient elution ginseng branch tuckahoe oral liquid, the method is simple to operate, reliable and stable, precision is high, and degree of separation is good, the stability of finger-print and reappearance better, and contain much information.
(2) due to the Precise levels that finger-print is not to measure certain composition, but the information of chemical composition to fully be reflected, therefore, the present invention selects to measure at 230nm wavelength place, and it is more that it goes out peak, and the information of reflection is more complete, each peak absorption value is good, and baseline is steady.
(3) the present invention adopts the quality control method of finger-print as ginseng branch tuckahoe oral liquid of ginseng branch tuckahoe oral liquid, both the one-sidedness judging preparation total quality because only measuring one, two chemical composition had been avoided, additionally reduce the possibility artificially processed for requisite quality, by carrying out systematic analysis to the sample of multiple batches, more comprehensive, scientifically can evaluate the quality of ginseng branch tuckahoe oral liquid, thus the quality of product and curative effect are guaranteed.
(4) the present invention adopts reference sample finger-print to carry out the comparison of finger-print, avoid each batch sample on the impact of reference fingerprint, strengthen the quality separating capacity of different batches sample, performance is better than reference fingerprint, and judged result more tallies with the actual situation.
Accompanying drawing explanation
Fig. 1 is each batch of ginseng branch tuckahoe oral liquid HPLC finger-print and reference fingerprint;
Fig. 2 is ginseng branch tuckahoe oral liquid reference substance mixed liquor finger-print;
Fig. 3 is the similarity sum (correlation coefficient process) of each batch sample and other batch sample;
Fig. 4 is the similarity (correlation coefficient process) of each batch sample and reference sample finger-print.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated, should be noted that following explanation is only to explain the present invention, not limiting its content.
Embodiment 1: the structure of ginseng branch tuckahoe oral liquid finger-print and Method validation
Concrete steps are:
(1) preparation of need testing solution: precision measures 1.25ml and joins branch tuckahoe oral liquid in 25ml volumetric flask, add methanol constant volume to scale, ultrasonic extraction 5min, place 0.5 hour in 4 DEG C of refrigerators, return to room temperature, through 0.22 μm of organic system membrane filtration, obtain the need testing solution of dilution 20 times, to obtain final product.
(2) preparation of reference substance solution:
Precision takes cinnamic acid standard items (lot number: 110786-200503) 0.0103g, puts in 100mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain cinnamic acid solution, and concentration is 0.103mg/mL.
Precision takes Paeoniflorin standard items (lot number: MUST-13030401) 0.0302g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain Paeoniflorin solution, and concentration is 3.02mg/mL.
Precision takes liquiritin standard items (lot number: 111610-200604) 0.0204g, puts in 25mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain liquiritin solution, and concentration is 0.816mg/mL.
Precision takes albiflorin standard items (lot number: MUST-14031214) 0.0101g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain albiflorin solution, and concentration is 1.01mg/mL.
Precision takes ammonium glycyrrhetate standard items (lot number: G07-110317) 0.0198g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain ammonium glycyrrhetate solution, and concentration is 1.98mg/mL.
(3) measure: accurate absorption need testing solution and reference substance solution respectively, inject high performance liquid chromatograph and measure, record chromatogram, and carry out methodological study.
Chromatographic condition is: chromatographic column is phenomenex synergi 4 μ, Hydro-RP chromatographic column (4 μ, 250 × 4.6mm); Mobile phase: 0.3% phosphoric acid water-acetonitrile, mobile phase A is 0.3% phosphate aqueous solution, and Mobile phase B is acetonitrile, type of elution: gradient elution; Determined wavelength 230nm, column temperature 30 DEG C, flow velocity 0.8ml/min, sample size 10 μ l, theoretical cam curve calculates by Paeoniflorin chromatographic peak and is not less than 5000, and be all greater than 1.0 with the degree of separation of other main chromatographic peak, all components has all been detected in 100min.Wherein, mobile phase linear gradient is as shown in table 1:
Table 1 chromatogram flow phase condition of gradient elution
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 95 | 5 |
| 5 | 95 | 5 |
| 10 | 92 | 8 |
| 20 | 80 | 20 |
| 25 | 77.5 | 22.5 |
| 40 | 67 | 33 |
| 50 | 60 | 40 |
| 60 | 50 | 50 |
| 80 | 15 | 85 |
| 90 | 5 | 95 |
| 100 | 95 | 5 |
Methodological study mainly comprise stability, reappearance and precision investigate, investigation method and result as follows:
1. stability experiment gets same batch of ginseng branch tuckahoe oral liquid sample, and after pretreatment, under the chromatographic condition optimized, analyze respectively at 0h, 2h, 4h, 8h, 12h, 16h and 24h sample introduction, record chromatogram, the results are shown in Table 2.From table 2, each main chromatographic peak relative retention time and its relative peak area ratio are without significant change, and RSD is respectively 0.050% ~ 0.31% and 2.0% ~ 4.3%, illustrates that the composition of need testing solution is stable in 24h.
2. Precision Experiment gets same batch of ginseng branch tuckahoe oral liquid sample, and after pretreatment, under the chromatographic condition optimized, METHOD FOR CONTINUOUS DETERMINATION 6 times, record chromatogram, the results are shown in Table 2.Visible, each main chromatographic peak relative retention time and its relative peak area ratio are without significant change, and RSD is respectively 0.01% ~ 0.25% and 1.7% ~ 4.6%, RSD < 5.0%, illustrates that the precision of instrument is good.
3. repeated experiment gets the ginseng branch tuckahoe oral liquid sample of 5 parts same batch, and after pretreatment, under the chromatographic condition optimized, sample introduction analysis respectively, record chromatogram, the results are shown in Table 2.Each main chromatographic peak relative retention time and its relative peak area ratio are without significant change, and RSD is respectively 0.01% ~ 1.2% and 0.8% ~ 4.6%, RSD < 5.0%, illustrates that the repeatability of this experimental technique is better.
Branch tuckahoe oral liquid fingerprint spectrum method the result joined by table 2
(4) with finger-print software, gained collection of illustrative plates is processed, branch tuckahoe oral liquid finger-print must be joined.Wherein, as shown in Figure 1, the finger-print of reference substance as shown in Figure 2 for the ginseng branch tuckahoe oral liquid sample of different batches and the finger-print of reference substance.
To join a tuckahoe oral liquid finger-print comparison visible with Fig. 1, need qualification and 5 kinds of quantitative component separating good, chromatographic peak peak shape symmetry, baseline is steady.The retention time of albiflorin, Paeoniflorin, liquiritin, cinnamic acid and ammonium glycyrrhetate is respectively: 25.2, and 26.2,30.4,47.2 and 55.7min.
Embodiment 2: utilize finger-print to carry out joining the quality control of branch tuckahoe oral liquid
Utilize finger-print to carry out joining the quality control of branch tuckahoe oral liquid, concrete grammar comprises the following steps:
(1) structure of ginseng branch tuckahoe oral liquid finger-print to be measured: get 11 batches of ginseng branch tuckahoe oral liquid samples to be measured, build the finger-print of ginseng branch tuckahoe oral liquid to be measured by the method for embodiment 1;
(2) the HPLC finger-print chromatographic peak of " similarity evaluation " software to 11 batches of ginseng branch tuckahoe oral liquid samples is adopted to mate.Select S1 (being numbered the sample of 1) as reference collection of illustrative plates, the reference fingerprint of generation is R.Between the finger-print of each batch sample and reference fingerprint and each sample, the similarity result of finger-print is in table 3.
The Similarity value (correlation coefficient process) of 11 batches, table 3 ginseng branch tuckahoe oral liquid finger-print
| S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | S9 | S10 | S11 | R | |
| S1 | 1.000 | 0.985 | 0.980 | 0.986 | 0.991 | 0.994 | 0.991 | 0.991 | 0.887 | 0.901 | 0.893 | 0.991 |
| S2 | 0.985 | 1.000 | 0.993 | 0.979 | 0.991 | 0.994 | 0.993 | 0.993 | 0.871 | 0.883 | 0.889 | 0.989 |
| S3 | 0.980 | 0.993 | 1.000 | 0.988 | 0.989 | 0.988 | 0.987 | 0.987 | 0.876 | 0.888 | 0.893 | 0.989 |
| S4 | 0.986 | 0.979 | 0.988 | 1.000 | 0.985 | 0.981 | 0.980 | 0.980 | 0.889 | 0.901 | 0.892 | 0.987 |
| S5 | 0.991 | 0.991 | 0.989 | 0.985 | 1.000 | 0.993 | 0.991 | 0.991 | 0.881 | 0.893 | 0.890 | 0.991 |
| S6 | 0.994 | 0.994 | 0.988 | 0.981 | 0.993 | 1.000 | 0.999 | 0.999 | 0.881 | 0.893 | 0.890 | 0.992 |
| S7 | 0.991 | 0.993 | 0.987 | 0.980 | 0.991 | 0.999 | 1.000 | 0.999 | 0.881 | 0.893 | 0.885 | 0.991 |
| S8 | 0.991 | 0.993 | 0.987 | 0.980 | 0.991 | 0.999 | 0.999 | 1.000 | 0.882 | 0.895 | 0.885 | 0.991 |
| S9 | 0.887 | 0.871 | 0.876 | 0.889 | 0.881 | 0.881 | 0.881 | 0.882 | 1.000 | 0.999 | 0.847 | 0.923 |
| S10 | 0.901 | 0.883 | 0.888 | 0.901 | 0.893 | 0.893 | 0.893 | 0.895 | 0.999 | 1.000 | 0.858 | 0.934 |
| S11 | 0.893 | 0.889 | 0.893 | 0.892 | 0.890 | 0.890 | 0.885 | 0.885 | 0.847 | 0.858 | 1.000 | 0.919 |
| R | 0.991 | 0.989 | 0.989 | 0.987 | 0.991 | 0.992 | 0.991 | 0.991 | 0.923 | 0.934 | 0.919 | 1.000 |
As can be seen from Table 3, the similarity of each batch sample and R, all more than 0.900, illustrates that the chemical composition of each batch sample of ginseng branch tuckahoe oral liquid is comparatively close generally.But be numbered 1 ~ 8 sample and the similarity of R comparatively large, more than 0.987, and the sample Similarity value of 9-11 less (0.919 ~ 0.934).From the Preliminary Analysis Results of similarity, 1 ~ 8 batch sample can be divided into a class, and 9 ~ 11 batch sample are another kind of.According to the information of each batch sample, 1 ~ 8 batch sample is 2013 ~ 2014 batch sample produced, and 9,10 batches is the sample of 2010, and 11 batches is the sample of 2012 years.The similarity of 9 ~ 11 batch sample and R is relatively low may be these batch sample time of making the product longlyer to cause in oral liquid caused by some composition or content change.The average collection of illustrative plates being employing 11 batch sample finger-prints due to reference fingerprint represents, therefore each batch sample finger-print all has a certain impact to the final R generated.Visible, the quality separating capacity of R to different batches sample is more weak.
In order to avoid each batch sample is on the impact of reference fingerprint, the present invention proposes the comparison carrying out finger-print based on reference sample finger-print.The construction method of reference sample finger-print is: the similarity first calculating each sample and other sample, then selects the sample maximum with other sample similarity sum to be reference sample.According to result of calculation, the similarity sum of the 6th batch sample and other batch sample is maximum (see Fig. 3), therefore selects the finger-print of the 6th batch sample as reference sample finger-print.Based on this sample, similarity evaluation is carried out to each batch sample, the results are shown in Figure 4.
As seen from Figure 4, the similarity of 1 ~ 8 batch sample and reference sample finger-print is all more than 0.981, and the similarity of 9 ~ 11 batch sample is 0.881 ~ 0.893, is less than 0.900.Illustrate that the quality of these batch sample is lower, consistance is bad.Compared with reference fingerprint (see table 3 data), the quality separating capacity of reference sample finger-print to different batches sample is enhanced, and performance is better than reference fingerprint, and judged result more tallies with the actual situation.Because this sample finger-print is for truly to measure finger-print, not by the impact of each batch sample finger-print, and maximum with other each batch sample fingerprint similarity sum, therefore there is good representativeness.
Claims (10)
1. the method for building up of a seed ginseng branch tuckahoe oral liquid finger-print, is characterized in that, comprise the following steps:
(1) preparation of need testing solution: get ginseng branch tuckahoe oral liquid, add methyl alcohol, ultrasonic extraction, 0.5-1 hour is placed in refrigeration, returns to room temperature, filters, gets subsequent filtrate, obtain need testing solution;
(2) preparation of reference substance solution: get cinnamic acid, Paeoniflorin, liquiritin, albiflorin, the ammonium glycyrrhetate reference substance of drying under reduced pressure to constant weight, add Methanol respectively and become reference substance solution;
(3) measure: precision measures need testing solution and reference substance solution respectively, inject high performance liquid chromatograph and measure, record chromatogram;
(4) with finger-print software, gained collection of illustrative plates is processed, branch tuckahoe oral liquid finger-print must be joined;
In step (3), the liquid phase chromatogram condition of mensuration is: chromatographic column is phenomenex synergi Hydro-RP chromatographic column; Mobile phase A is 0.3% phosphate aqueous solution, and Mobile phase B is acetonitrile; Gradient elution; Flow rate of mobile phase is 0.8ml/min; Column temperature is 30 DEG C; Determined wavelength is 230nm.
2. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, is characterized in that, in step (3), in gradient elution process, being changed to of mobile phase A and Mobile phase B: 0-5min, mobile phase A 95%-95%, Mobile phase B 5%-5%; 5-10min, mobile phase A 95%-92%, Mobile phase B 5%-8%; 10-20min, mobile phase A 92%-80%, Mobile phase B 8%-20%; 20-25min, mobile phase A 80%-77.5%, Mobile phase B 20%-22.5%; 25-40min, mobile phase A 77.5%-67%, Mobile phase B 22.5%-33%; 40-50min, mobile phase A 67%-60%, Mobile phase B 33%-40%; 50-60min, mobile phase A 60%-50%, Mobile phase B 40%-50%; 60-80min, mobile phase A 50%-15%, Mobile phase B 50%-85%; 80-90min, mobile phase A 15%-5%, Mobile phase B 85%-95%; 90-100min, mobile phase A 5%-95%, Mobile phase B 95%-5%.
3. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, it is characterized in that, in step (1), the time that refrigeration is placed is 0.5-1 hour.
4. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, it is characterized in that, in step (1), the preparation method of need testing solution is: precision measures 1.25ml and joins branch tuckahoe oral liquid in 25ml volumetric flask, adds methanol constant volume to scale, ultrasonic extraction 5min, place 0.5 hour in 4 DEG C, return to room temperature, through 0.22 μm of organic system membrane filtration, obtain need testing solution.
5. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, it is characterized in that, in step (2), the preparation method of reference substance solution is:
Precision takes cinnamic acid standard items 0.0103g, puts in 100mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain cinnamic acid solution, and concentration is 0.103mg/mL;
Precision takes Paeoniflorin standard items 0.0302g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain Paeoniflorin solution, and concentration is 3.02mg/mL;
Precision takes liquiritin standard items 0.0204g, puts in 25mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain liquiritin solution, and concentration is 0.816mg/mL;
Precision takes albiflorin standard items 0.0101g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain albiflorin solution, and concentration is 1.01mg/mL;
Precision takes ammonium glycyrrhetate standard items 0.0198g, puts in 10mL volumetric flask, adds methyl alcohol and dissolves and be diluted to scale, obtain ammonium glycyrrhetate solution, and concentration is 1.98mg/mL.
6. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, it is characterized in that, in step (3), the packing material size of described chromatographic column is 4 μm, and column length is 250mm, and column internal diameter is 4.6mm.
7. the method for building up of ginseng branch tuckahoe oral liquid finger-print as claimed in claim 1, it is characterized in that, in step (3), the accurate volume measuring need testing solution and reference substance solution is 10 μ L respectively.
8. method according to claim 1 builds the ginseng branch tuckahoe oral liquid finger-print obtained, and it is characterized in that, the total peak in described finger-print is 11.
9. the method for quality control of a seed ginseng branch tuckahoe oral liquid, is characterized in that, comprise the following steps:
(1) structure of ginseng branch tuckahoe oral liquid finger-print to be measured: get ginseng branch tuckahoe oral liquid to be measured, build the finger-print of ginseng branch tuckahoe oral liquid to be measured by method according to claim 1;
(2) finger-print of ginseng branch tuckahoe oral liquid to be measured and reference sample finger-print are carried out similarity system design, to evaluate the quality of ginseng branch tuckahoe oral liquid to be measured, computing method are correlation coefficient process, similarity be not less than 0.900 oral liquid up-to-standard.
10. the method for quality control of ginseng branch tuckahoe oral liquid as claimed in claim 9, it is characterized in that, in step (2), the construction method of described reference sample finger-print is: the similarity first calculating each sample and other sample, then select the sample maximum with other sample similarity sum to be reference sample, its finger-print is reference sample finger-print.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510224392.4A CN104849364B (en) | 2015-05-05 | 2015-05-05 | Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510224392.4A CN104849364B (en) | 2015-05-05 | 2015-05-05 | Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104849364A true CN104849364A (en) | 2015-08-19 |
| CN104849364B CN104849364B (en) | 2017-02-22 |
Family
ID=53849163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510224392.4A Active CN104849364B (en) | 2015-05-05 | 2015-05-05 | Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104849364B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105259295A (en) * | 2015-11-17 | 2016-01-20 | 山东沃华医药科技股份有限公司 | Quality detection method for ginseng, cassia twig and poria cocos oral solution |
| CN105467052A (en) * | 2015-11-18 | 2016-04-06 | 江苏康缘药业股份有限公司 | A component detecting method for a Sen Wu kidney-tonifying tablet and a fingerprint constructing method |
| CN107632086A (en) * | 2017-09-12 | 2018-01-26 | 山东大学 | The construction method of one seed ginseng branch tuckahoe oral liquid finger-print and application |
| CN108107130A (en) * | 2017-12-22 | 2018-06-01 | 山东沃华医药科技股份有限公司 | The assay method of one seed ginseng branch Siberian cocklebur preparation finger |
| CN108459129A (en) * | 2017-02-17 | 2018-08-28 | 华润三九医药股份有限公司 | A kind of method of quality control of Tetrandra and Poria Decoction composition |
| CN110057927A (en) * | 2018-12-28 | 2019-07-26 | 山东大学 | A kind of detection method and its application of Qilong Capsule finger-print combination LC-MS |
| CN112782326A (en) * | 2020-12-30 | 2021-05-11 | 上海交通大学 | Method for simultaneously measuring fingerprint spectrum and multi-index component content of Weikangling capsule |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102114094A (en) * | 2010-01-01 | 2011-07-06 | 江苏康缘药业股份有限公司 | Chromatographic fingerprint of cassia twig and Tuckahoe pharmaceutical composition and detection method and application thereof |
-
2015
- 2015-05-05 CN CN201510224392.4A patent/CN104849364B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102114094A (en) * | 2010-01-01 | 2011-07-06 | 江苏康缘药业股份有限公司 | Chromatographic fingerprint of cassia twig and Tuckahoe pharmaceutical composition and detection method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| WEIDONG PAN等: "Shen-Zhi-Ling Oral Liquid Improves Behavioral and Psychological Symptoms of Dementia in Alzheimer’s Disease", 《EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE》 * |
| 何超元: "近红外光谱应用于参枝苓口服液中肉桂酸和芍药苷含量的快速测定研究", 《中国优秀硕士学位论文全文数据库工程科技I辑》 * |
| 李毛等: "参枝苓口服液对阿尔茨海默病患者精神行为症状的影响", 《上海中医药大学学报》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105259295A (en) * | 2015-11-17 | 2016-01-20 | 山东沃华医药科技股份有限公司 | Quality detection method for ginseng, cassia twig and poria cocos oral solution |
| CN105259295B (en) * | 2015-11-17 | 2017-05-17 | 山东沃华医药科技股份有限公司 | Quality detection method for ginseng, cassia twig and poria cocos oral solution |
| CN105467052A (en) * | 2015-11-18 | 2016-04-06 | 江苏康缘药业股份有限公司 | A component detecting method for a Sen Wu kidney-tonifying tablet and a fingerprint constructing method |
| CN105467052B (en) * | 2015-11-18 | 2017-03-29 | 江苏康缘药业股份有限公司 | The black kidney tonifying piece component detection method of ginseng and fingerprint map construction method |
| CN108459129A (en) * | 2017-02-17 | 2018-08-28 | 华润三九医药股份有限公司 | A kind of method of quality control of Tetrandra and Poria Decoction composition |
| CN108459129B (en) * | 2017-02-17 | 2020-06-05 | 华润三九医药股份有限公司 | Quality control method of radix Stephaniae Tetrandrae and Poria decoction composition |
| CN107632086A (en) * | 2017-09-12 | 2018-01-26 | 山东大学 | The construction method of one seed ginseng branch tuckahoe oral liquid finger-print and application |
| CN108107130A (en) * | 2017-12-22 | 2018-06-01 | 山东沃华医药科技股份有限公司 | The assay method of one seed ginseng branch Siberian cocklebur preparation finger |
| CN110057927A (en) * | 2018-12-28 | 2019-07-26 | 山东大学 | A kind of detection method and its application of Qilong Capsule finger-print combination LC-MS |
| CN110057927B (en) * | 2018-12-28 | 2020-04-21 | 山东大学 | Detection method combining fingerprint spectrum and liquid chromatography-mass spectrometry of Qilong capsules and application of detection method |
| CN112782326A (en) * | 2020-12-30 | 2021-05-11 | 上海交通大学 | Method for simultaneously measuring fingerprint spectrum and multi-index component content of Weikangling capsule |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104849364B (en) | 2017-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104849364A (en) | Canzhiling oral solution fingerprint map building method, fingerprint map and application thereof | |
| CN102138985B (en) | Quality control method of total glycosides single preparation of white paeony roots | |
| CN109709251B (en) | Detection method of fingerprint of poria, cassia, rhizoma atractylodis and licorice decoction | |
| CN111337589A (en) | Method for establishing orange-shell mixture HPLC fingerprint spectrum | |
| CN106198823B (en) | A kind of assay method of Shenshuaining piece active ingredient | |
| CN111830160B (en) | Method for detecting fingerprint spectrum of Qijudihuang pills and application thereof | |
| CN105842353A (en) | Establishing method of fingerprint spectrum of honeysuckle-fructus forsythiae heat-clearing tablets and fingerprint spectrum | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN109374764B (en) | HPLC fingerprint spectrum and main component content determination method for eight-ingredient Longzui granules | |
| CN107356691A (en) | Build the detection method of bent finger-print | |
| CN104764820A (en) | Method for determining content of active ingredients such as ephedrine hydrochloride and pseudoephedrine hydrochloride in pinellia ternata syrup | |
| CN104849375B (en) | The detection method of 'Juhong Tanke ' | |
| CN114487242B (en) | Characteristic spectrum of endothelium corneum Gigeriae Galli and/or vinegar endothelium corneum Gigeriae Galli and its preparation, and its construction method and content determination method | |
| CN104316613A (en) | Method for establishing fingerprint spectrum of wind-dispelling and detoxifying capsules | |
| CN104007198B (en) | A kind of glossy ganoderma emperor's preparation HPLC standard finger-print and construction method thereof and application | |
| CN105675761A (en) | HPLC method for simultaneously determining contents of four alkaloid components in cortex berberidis | |
| CN109709222B (en) | Component detection method of Ganmaoling and compound Ganmaoling | |
| CN107807190A (en) | A kind of HPLC methods of 3 kinds of active components in measure hymsleya amabilis | |
| CN105510452A (en) | Multiple index component content determination, fingerprint building and preparation methods of liver-tonifying eyesight-improving oral liquid | |
| CN109374758A (en) | The quantitative finger print atlas and its detection method of phellodendron extract and application | |
| CN109781884B (en) | Establishing method of Qianliexin capsule fingerprint and fingerprint thereof | |
| CN104849384A (en) | Method for establishing fingerprint spectrum of Jian Ganle preparation | |
| CN110308213B (en) | Method for constructing fingerprint spectrum of kidney-nourishing and fetus-growing pill and application of fingerprint spectrum in quality detection | |
| CN110441414B (en) | Method for establishing fingerprint of Jinshui Liujun decoction | |
| CN103163272A (en) | Quality control method of infant spleen tonifying medicament |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |