CN106018647A - Method for establishing sunflower plate HPLC standard fingerprint - Google Patents
Method for establishing sunflower plate HPLC standard fingerprint Download PDFInfo
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- CN106018647A CN106018647A CN201610541603.1A CN201610541603A CN106018647A CN 106018647 A CN106018647 A CN 106018647A CN 201610541603 A CN201610541603 A CN 201610541603A CN 106018647 A CN106018647 A CN 106018647A
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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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
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Abstract
The invention relates to a method for establishing a sunflower plate HPLC standard fingerprint. The method includes the steps of detecting multiple batches of sunflower plate extraction solutions through an HPLC method to obtain a fingerprint, and analyzing the fingerprint through software to obtain the sunflower plate standard fingerprint. In the standard fingerprint establishing process, great attention is paid to the sequence and relation of all characteristic peaks forming the fingerprint, and attention is paid to the overall appearance characteristics; by optimizing the sunflower plate extract preparation method and the HPLC liquid phase conditions, the obtained standard fingerprint can effectively and accurately express the quality of a sunflower plate, the quality of the sunflower plate can be easily and comprehensively monitored, and the aim of rapidly and accurately identifying the authenticity and quality of the product is achieved.
Description
Technical field
The invention belongs to technical field of analytical chemistry, particularly relate to a kind of set up building of Receptaculum Helianthi HPLC standard finger-print
Cube method.
Background technology
Chinese crude drug yield is fewer and feweri now, anxious new material to be developed.Receptaculum Helianthi be Helianthi slough after seed to
Day certain herbaceous plants with big flowers dish, is the side-product of Helianthi, and yield accounts for the 20% of the Helianthi yield of kernels, in Helianthi main producing region be one huge
Resource.
Receptaculum Helianthi controls headache, blurred vision, toothache, stomach, stomachache, woman month dysmenorrhoea, and skin ulcer swells.Pharmacological action shows the second of Receptaculum Helianthi
Alcohol extract, to anesthetized cat intravenous injection, can be significantly reduced blood pressure effect.Clinical application shows, takes Receptaculum Helianthi appropriate, adds
Decocting becomes paste, and external application has certain curative effect to innominate toxic swelling, rheumatic arthritis, scapulohumeral periarthritis；Take Receptaculum Helianthi to dry and cut
Broken, parching to brown, grind into powder.Take after mixing it with water with Chinese liquor or boiled water sugaring, can be used for treating mastitis；With Receptaculum Helianthi 3 liang and Herba Pteridis Multifidae 2 liang, water
2 liang of Fructus Myricae rubrae (herb), decocting 1～2 hours one-tenth half are gelatin, oral, have certain curative effect to tumor；And have zoopery to demonstrate,prove
Bright, Receptaculum Helianthi alcohol leaching liquid has significant hypotensive effect.
Fingerprint pattern technology has become that internationally recognized control is important or one of the most effective means of natural drug quality, it
Can more comprehensively, accurately, reaction Multiple components distribution in Chinese medicine or its preparation intuitively, be a kind of comprehensive, macroscopic view,
Quantifiable discrimination method.Use fingerprint pattern technology, by the identification at principal character peak in spectrogram and/or the survey of ratio
Fixed, can effectively differentiate the true and false of medical material.But, about the fingerprint chromatogram of Receptaculum Helianthi, there is not been reported.
Therefore, it is established that the fingerprint spectrum method of Receptaculum Helianthi, significant for evaluating the quality of Receptaculum Helianthi.
Summary of the invention
First purpose of the present invention is to provide a kind of method setting up Receptaculum Helianthi HPLC standard finger-print, specifically uses
Following technical scheme:
A kind of method setting up Receptaculum Helianthi HPLC standard finger-print, including using HPLC method to 5-15 batch sunflower
The extract of dish carries out detection and obtains the step of finger printing, and described HPLC method is with octadecylsilane chemically bonded silica as chromatograph
Column packing, with acetonitrile for mobile phase A phase, 0.2-0.6% phosphate aqueous solution is Mobile phase B phase, is carried out as follows gradient
Eluting:
Wherein, in described gradient elution, 30～35min, " 9 → 80 " represent and are meant that, the ratio of mobile phase A is by 9%
Being gradually increased to 80%, " 91 → 20 " represent and are meant that, the ratio of Mobile phase B is gradually decrease to 20% by 91%；Other time
Between the implication of section be similar to therewith.
During standard finger-print determines, to fully focus on each and constitute order before and after fingerprint characteristic peak, mutually close
System, and overall facial feature, inventor has carried out numerous studies to the preparation method of Receptaculum Helianthi extract, finds extract
Preparation method has the biggest impact to the form of ultimate criterion finger printing, specifically, when only extracting by water method, deposits
Extracting, the composition obtained is too much, and obtained collection of illustrative plates chromatographic peak is too much, the problem that can not efficiently separate between peak and peak；When
Time only with alcohol extraction, there is fractions can not effectively be extracted, and it is very few that obtained collection of illustrative plates also exists chromatographic peak
Problem.Additionally, extract component is not the most used in clothes for patients use, extract component is crossed and can not be reached therapeutic purposes at least.
Inventor, through research, finally determines the extracting method of following Receptaculum Helianthi extract, it may be assumed that first use water extraction sunflower
Dish, then with the extracting solution of ethanol precipitation gained, takes supernatant concentration and becomes thick paste, dry, pulverize, to obtain final product.
The material obtained to adopt this method extraction detects as test sample, the standard finger-print finally given
In can efficiently separate between peak before and after each characteristic fingerprint peak, meet examination requirements, this standard finger-print can be more accurate
Ground characterizes the quality of Receptaculum Helianthi, the beneficially quality of overall monitor product, and the quality testing for medicine provides reliable foundation.And
Method of the present invention has simple to operate, the satisfactory feature of separating degree between chromatographic peak.
Preferably, the extract of described Receptaculum Helianthi extracts by the following method and obtains: pulverizing Receptaculum Helianthi to particle diameter is 0.5-
1.5cm, adds the water being equivalent to Receptaculum Helianthi weight 8-10 times, extracts 2-4 time, each 1-3h, concentrated extracting solution, concentrates to gained
Liquid adds ethanol precipitation, centrifugal, take supernatant, remove ethanol and be condensed into thick paste, dry, pulverize, to obtain final product.
Obtain it is further preferred that the extract of described Receptaculum Helianthi extracts by the following method: take Receptaculum Helianthi, be crushed to grain
Footpath is 1cm, is separately added into the water being equivalent to Receptaculum Helianthi weight 10,8,8 times, extracts 3 times, each 2 hours；Filter respectively, merge
Filtrate, is concentrated into relative density 1.06 (20 DEG C), adds ethanol and makes alcohol content be 65%, stands 24 hours, centrifugal, takes supernatant,
Remove ethanol and be condensed into thick paste, dry, pulverize, to obtain final product.
Wherein, described " alcohol content " ethanol referred to percentage by volume in the solution.
HPLC method of the present invention uses UV-detector, and detection wavelength is 320-330nm；And/or, flow velocity is
0.8-1.2mL/min；And/or, column temperature is 28-32 DEG C.
Preferably, detection wavelength is 327nm, and flow velocity is 1.0mL/min, and column temperature is 30 DEG C.
Most preferably, the present invention sets up the method for described standard finger-print and comprises the steps:
(1) preparation of need testing solution: weigh the Receptaculum Helianthi extract of 5-15 batch, respectively with methanol as solvent, join
Make the need testing solution that concentration is 10-50mg/mL；
(2) HPLC measures: uses following HPLC method to detect 5-15 described need testing solution, obtains fingerprint image
Spectrum；
With octadecylsilane chemically bonded silica for chromatographic column filler, with acetonitrile for mobile phase A phase, 0.4% phosphoric acid is water-soluble
Liquid is Mobile phase B phase, is carried out as follows gradient elution；Ultraviolet detection wavelength is 327nm, and flow velocity is 1.0mL/min, column temperature
It it is 30 DEG C；
(3) foundation of standard finger-print: use software that the finger printing of step (2) is analyzed, obtain Receptaculum Helianthi
Standard finger-print.
Wherein, the number of described batch is preferably 10.
Described software can use software commonly used in the art, present invention preferably employs the Chinese medicine that Chinese Pharmacopoeia committee is recommended
Chromatographic fingerprinting similarity evaluation systems soft ware.
Second object of the present invention is to provide any one method above-mentioned and sets up the Receptaculum Helianthi HPLC standard fingerprint obtained
Collection of illustrative plates.
Described standard finger-print is made up of 9 total peaks, according to the ascending sequence of retention time, each total peak and
The relative retention time at the peak that 7 retention times are corresponding is respectively as follows: No. 1 peak: 0.16 ± 5%；No. 2 peak: 0.21 ± 5%；
No. 3 peak: 0.29 ± 5%；No. 4 peak: 0.52 ± 5%；No. 5 peak: 0.55 ± 5%；No. 6 peak: 0.79 ± 5%；No. 7
Peak: 1.00；No. 8 peak: 1.21 ± 5%；No. 9 peak: 1.28 ± 5%.
Wherein, described 7th peak that retention time is corresponding is chlorogenic acid peak.The present invention using chlorogenic acid as reference substance, with
The peak that finger printing Content of Chlorogenic Acid is corresponding is with reference to peak, calculates other total peaks relative retention time relative to chlorogenic acid peak.
Finger printing Content of Chlorogenic Acid peak is positioned by reference substance solution, and the compound method of wherein said reference substance solution is by green former
Acid is dissolved in methanol, is configured to the solution that concentration is 10-30 μ g/mL, uses the HPLC method identical with measuring need testing solution
It is measured.
Third object of the present invention is to provide a kind of method detecting Receptaculum Helianthi quality, particularly as follows: first, sets up sunflower
Dish HPLC standard finger-print；Secondly, use the HPLC method identical with Criterion finger printing that Receptaculum Helianthi to be measured is carried
Take thing to detect, obtain finger printing；Finally, described finger printing and Receptaculum Helianthi HPLC standard fingerprint spectrogram are carried out point
Analysis, the two similarity is qualified products more than 0.95, is substandard product less than or equal to 0.95.
Wherein, the method for building up of Receptaculum Helianthi HPLC standard finger-print can be carried out according to any one method above-mentioned.To be measured
The extract of Receptaculum Helianthi can extract according to the method described in any one above.
Most preferably, the method for detection Receptaculum Helianthi quality comprises the steps:
(1) preparation of need testing solution: pulverizing Receptaculum Helianthi to particle diameter is 0.5-1.5cm, adds and is equivalent to Receptaculum Helianthi weight
The water of 8-10 times, extracts 2-4 time, each 1-3h, concentrated extracting solution, adds ethanol precipitation in gained concentrated solution, centrifugal, takes
Clear liquid, removes ethanol and is condensed into thick paste, dry, pulverize, obtain Receptaculum Helianthi extract；First is added in described Receptaculum Helianthi extract
Alcohol, is configured to the need testing solution that concentration is 10-50mg/mL；
(2) HPLC measures: use the confession of HPLC method 5-15 the batch to preparing according to step (1) described method
Test sample solution detects, and obtains finger printing；
Described HPLC method is with octadecylsilane chemically bonded silica for chromatographic column filler, with acetonitrile for mobile phase A phase,
0.2%-0.6% phosphate aqueous solution is Mobile phase B phase, is carried out as follows gradient elution:
(3) foundation of standard finger-print: use software that the finger printing of step (2) is analyzed, generate Receptaculum Helianthi
HPLC standard finger-print；
(4) acquisition of testing sample finger printing: according to the operation of step (1), Receptaculum Helianthi to be measured is processed, produce
Testing sample solution, detects described testing sample solution according to the HPLC method identical with step (2), obtains sunflower to be measured
The finger printing of dish；
(5) Quality estimation: use software that finger printing and the Receptaculum Helianthi HPLC standard fingerprint spectrogram of Receptaculum Helianthi to be measured are entered
Row is analyzed, and similarity is qualified products more than 0.95, is substandard product less than or equal to 0.95.
Wherein, described software is preferably chromatographic fingerprints of Chinese materia medica similarity evaluation systems soft ware.
The quality determining method of the present invention can effectively monitor the quality of Receptaculum Helianthi, it is ensured that its quality stable, homogeneous,
Controlled, there is method simplicity, good stability, precision height, high repeatability and other advantages, the true of product can be differentiated quickly and accurately
Pseudo-good and bad.
Accompanying drawing explanation
Fig. 1 is the liquid chromatogram of chlorogenic acid；
Fig. 2 is the liquid chromatogram of 10 batch Receptaculum Helianthi extracts；
Fig. 3 is Receptaculum Helianthi HPLC standard finger-print；
In Fig. 1-3, abscissa is response time (min), and vertical coordinate is response intensity (unit: UV).
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.
The extract of the Receptaculum Helianthi related in following example extracts the most by the following method and obtains: take sunflower
Dish, being crushed to particle diameter is 1cm, is separately added into the water being equivalent to Receptaculum Helianthi weight 10,8,8 times, extracts 3 times, each 2 hours, divides
Do not filter, merging filtrate, be concentrated into relative density 1.06 (20 DEG C), add ethanol and make alcohol content be 65%, stand 24 hours, from
The heart, takes supernatant, removes ethanol and is condensed into thick paste, dry, pulverize, to obtain final product.
In embodiment use chromatograph of liquid be Shimadzu LC-20A high performance liquid chromatograph (binary geopressure gradient pump,
Line degasser, column oven (column temperature 30 DEG C), automatic sampler, UV-detector).
The chlorogenic acid reference substance used identifies institute purchased from China's pharmaceutical biological product, and liquid-phase chromatographic analysis reagent is chromatograph
Pure, remaining reagent is analytical pure, and water is ultra-pure water.
The foundation of embodiment 1 Receptaculum Helianthi HPLC standard finger-print
(1) preparation of need testing solution: weigh Receptaculum Helianthi extract 0.55g, accurately weighed, put in 50ml measuring bottle, add first
Alcohol 40ml, supersound process (power 300W, frequency 25kHz) 45 minutes, let cool, add methanol to scale, shake up, filter, take continuous filter
Liquid, obtains (keeping in Dark Place).
(2) HPLC measures: uses following HPLC method to detect 10 described need testing solutions, obtains finger printing
(as shown in Figure 2)；
Chromatographic column is Agilent ZORBAX SB-C18(5 μm, 4.6 × 250mm), with acetonitrile for mobile phase A phase, 0.4%
Phosphate aqueous solution is Mobile phase B phase, is carried out as follows gradient elution；Ultraviolet detection wavelength is 327nm, and flow velocity is 1.0mL/
Min, column temperature is 30 DEG C；
(3) foundation of standard finger-print: use chromatographic fingerprints of Chinese materia medica similarity evaluation systems soft ware to step (2)
Finger printing be analyzed, determine common characteristic peak, obtain the standard finger-print (as shown in Figure 3) of Receptaculum Helianthi.
The standard finger-print of the application is using chlorogenic acid as with reference to product, and wherein, chlorogenic acid reference substance solution is by green former
Acid is dissolved in methanol and is configured to the solution that concentration is 20 μ g/mL, then uses the condition of above-mentioned steps (2) to detect, obtains
The liquid chromatogram (as shown in Figure 1) of reference substance.Then, with standard finger-print Content of Chlorogenic Acid peak for reference to peak, being calculated
Other each total peaks relative retention time relative to chlorogenic acid peak.Each total peak in Receptaculum Helianthi HPLC standard finger-print
Retention time, relative retention time and peak area are as shown in table 1:
The total retention time at peak, relative retention time and peak area in table 1 Receptaculum Helianthi HPLC standard finger-print
The quality evaluating method of embodiment 2 Receptaculum Helianthi
(1) preparation of testing sample solution: take 3 batches of Receptaculum Helianthi to be measured, prepare Receptaculum Helianthi to be measured according to preceding method
Extract, takes 1g Receptaculum Helianthi extract respectively, to tool plug conical flask, adds methanol 25mL, weighed weight, supersound process (power
250W, frequency 25kHz) 45 minutes, let cool, more weighed weight, add methanol and supply the weight of less loss, shake up, filter, take continuous filter
Liquid, obtains testing sample solution (keeping in Dark Place)；
(2) acquisition of testing sample finger printing: use the HPLC method identical with embodiment 1 step (2) to step (1)
3 samples detect, obtain finger printing；
(3) Quality estimation: use the standard that embodiment 1 is obtained by chromatographic fingerprints of Chinese materia medica similarity evaluation systems soft ware
The finger printing of finger printing and testing sample is analyzed, and similarity is respectively 0.985,0.994,0.989, is qualified product
Product.
Evaluation of methodology method
Precision
Experimental technique: the preparation of reference substance solution: take chlorogenic acid reference substance 7.027mg, accurately weighed, put 100ml measuring bottle
In, adding methanol and make dissolving in right amount and be diluted to scale, shake up, precision measures 2.5ml, puts in 10ml measuring bottle, with methanol dilution extremely
Scale.Being carried out continuously 6 times, record chromatogram, result is as shown in table 2.
Table 2: Precision Experiment result
Experimental result: from above-mentioned test, this analysis method, good to the assay Intermediate precision of chlorogenic acid,
Meet its testing requirement.
Stability
Experimental technique: the preparation of need testing solution: weigh Receptaculum Helianthi extract 0.55g, accurately weighed, put 50ml measuring bottle
In, add methanol 40ml, supersound process (power 300W, frequency 25kHz) 45 minutes, let cool, add methanol to scale, shake up, filter,
Take subsequent filtrate, obtain (keeping in Dark Place).
Respectively at 0 hour, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, within 24 hours, draw 10 μ l and inject liquid
In chromatography, record chromatogram.Result is as shown in table 3:
Table 3: solution stability testing result
Experimental result: from the foregoing, it will be observed that Receptaculum Helianthi powder Content of Chlorogenic Acid peak area is had good stability by this analysis method.
Repeatability
Experimental technique: the preparation of reference substance solution: take chlorogenic acid reference substance 7.027mg, accurately weighed, put 100ml measuring bottle
In, adding methanol and make dissolving in right amount and be diluted to scale, shake up, precision measures 2.5ml, puts in 10ml measuring bottle, with methanol dilution extremely
Scale.
The preparation of need testing solution: take content under content uniformity item, mixing, weigh 0.55g, accurately weighed, put 50ml
In measuring bottle, add methanol 40ml, supersound process (power 300W, frequency 25kHz) 45 minutes, let cool, add methanol to scale, shake up,
Filter, take subsequent filtrate, obtain (keeping in Dark Place).
Precision draws need testing solution and each 10 μ l of reference substance solution respectively, injects chromatograph of liquid, measures, and records chromatograph
Figure.Result is as shown in table 4:
Table 4: repeated experiment result
Experimental result: from above-mentioned test, this analysis method, good to the peak area repeatability of Receptaculum Helianthi powder Content of Chlorogenic Acid
Good, meet its testing requirement.
Although, used general explanation, detailed description of the invention and test, the present invention made detailed retouching
Stating, but on the basis of the present invention, can make some modifications or improvements it, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Scope.
Claims (10)
1. the method setting up Receptaculum Helianthi HPLC standard finger-print, it is characterised in that: include using HPLC method to 5-15
The extract of batch Receptaculum Helianthi carries out detection and obtains the step of finger printing；
Described HPLC method is with octadecylsilane chemically bonded silica for chromatographic column filler, with acetonitrile for mobile phase A phase, 0.2%-
0.6% phosphate aqueous solution is Mobile phase B phase, is carried out as follows gradient elution:
。
Method the most according to claim 1, it is characterised in that the extract of described Receptaculum Helianthi extracts by the following method
Arrive: use water extraction Receptaculum Helianthi, with the extracting solution of ethanol precipitation gained, take supernatant concentration and become thick paste, dry, pulverize, to obtain final product.
Method the most according to claim 1 and 2, it is characterised in that: the extract of described Receptaculum Helianthi carries by the following method
Acquirement to: pulverizing Receptaculum Helianthi to particle diameter is 0.5-1.5cm, adds and is equivalent to the water of Receptaculum Helianthi weight 8-10 times, extracts 2-4 time,
1-3h every time, concentrated extracting solution, in gained concentrated solution, add ethanol precipitation, centrifugal, take supernatant, remove ethanol and be condensed into thick
Cream, dry, pulverize, and to obtain final product；
Preferably, adding ethanol makes its volumn concentration in the solution be 60-70%.
4. according to the method described in any one of claim 1-3, it is characterised in that: described HPLC method uses UV-detector,
Detection wavelength is 320-330nm；And/or, flow velocity is 0.8-1.2mL/min；And/or, column temperature is 28-32 DEG C.
5. according to the method described in any one of claim 1-4, it is characterised in that comprise the steps:
(1) preparation of need testing solution: weigh the extract of 5-15 batch Receptaculum Helianthi, respectively with methanol as solvent, is configured to dense
Degree is the need testing solution of 10-50mg/mL；
(2) HPLC measures: uses following HPLC method to detect 5-15 described need testing solution, obtains finger printing；
With octadecylsilane chemically bonded silica for chromatographic column filler, with acetonitrile for mobile phase A phase, 0.4% phosphate aqueous solution is
Mobile phase B phase, is carried out as follows gradient elution；Ultraviolet detection wavelength is 327nm, and flow velocity is 1.0mL/min, and column temperature is 30
℃；
(3) foundation of standard finger-print: use software that the finger printing of step (2) is analyzed, obtain the mark of Receptaculum Helianthi
Quasi-finger printing；
Preferably, described software is chromatographic fingerprints of Chinese materia medica similarity evaluation systems soft ware.
6. according to the method described in any one of claim 1-5, it is characterised in that the quantity of described batch is 10.
7. the method described in any one of claim 1-6 sets up the Receptaculum Helianthi HPLC standard finger-print obtained, it is characterised in that
Described Receptaculum Helianthi HPLC standard finger-print is made up of 9 total peaks, according to the ascending sequence of retention time, each total peak with
The relative retention time at the 7th peak that retention time is corresponding is respectively as follows: No. 1 peak: 0.16 ± 5%；No. 2 peak: 0.21 ±
5%；No. 3 peak: 0.29 ± 5%；No. 4 peak: 0.52 ± 5%；No. 5 peak: 0.55 ± 5%；No. 6 peak: 0.79 ± 5%；
No. 7 peak: 1.00；No. 8 peak: 1.21 ± 5%；No. 9 peak: 1.28 ± 5%.
Receptaculum Helianthi HPLC standard finger-print the most according to claim 7, it is characterised in that: described No. 7 peak is corresponding
Material is chlorogenic acid.
9. the method detecting Receptaculum Helianthi quality, it is characterised in that: first, set up Receptaculum Helianthi HPLC standard finger-print；Its
Secondary, use the HPLC method identical with Criterion finger printing that the extract of Receptaculum Helianthi to be measured is detected, obtain fingerprint
Collection of illustrative plates；Finally, being analyzed described finger printing and Receptaculum Helianthi HPLC standard fingerprint spectrogram, the two similarity more than 0.95 is
Qualified products, are substandard product less than or equal to 0.95.
Method the most according to claim 9, it is characterised in that comprise the steps:
(1) preparation of need testing solution: pulverizing Receptaculum Helianthi to particle diameter is 0.5-1.5cm, adds and is equivalent to Receptaculum Helianthi weight 8-10
Water again, extracts 2-4 time, each 1-3h, concentrated extracting solution, adds ethanol precipitation in gained concentrated solution, centrifugal, takes supernatant
Liquid, removes ethanol and is condensed into thick paste, dry, pulverize, obtain Receptaculum Helianthi extract；First is added in described Receptaculum Helianthi extract
Alcohol, is configured to the need testing solution that concentration is 10-50mg/mL；
(2) HPLC measures: use the test sample of HPLC method 5-15 the batch to preparing according to step (1) described method
Solution detects, and obtains finger printing；
Described HPLC method is with octadecylsilane chemically bonded silica for chromatographic column filler, with acetonitrile for mobile phase A phase, 0.2%-
0.6% phosphate aqueous solution is Mobile phase B phase, is carried out as follows gradient elution:
(3) foundation of standard finger-print: use software that the finger printing of step (2) is analyzed, generate Receptaculum Helianthi HPLC
Standard finger-print；
(4) acquisition of testing sample finger printing: according to the operation of step (1), Receptaculum Helianthi to be measured is processed, produce to be measured
Sample solution, detects described testing sample solution according to the HPLC method identical with step (2), obtains Receptaculum Helianthi to be measured
Finger printing；
(5) Quality estimation: use software the finger printing of Receptaculum Helianthi to be measured and Receptaculum Helianthi HPLC standard fingerprint spectrogram to be carried out point
Analysis, similarity is qualified products more than 0.95, is substandard product less than or equal to 0.95；
Preferably, described software is chromatographic fingerprints of Chinese materia medica similarity evaluation systems soft ware.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110501200A (en) * | 2019-09-23 | 2019-11-26 | 吉林师范大学 | A kind of black nightshade effective ingredient that can be recycled is extracted and separation method |
| CN115290805A (en) * | 2022-08-23 | 2022-11-04 | 陕西省中医药研究院 | Finger print method and content detection method for radix Pteris Multifidae medicinal material |
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