CN110361459A - The method for building up of loguat leaf standard decoction finger-print - Google Patents
The method for building up of loguat leaf standard decoction finger-print Download PDFInfo
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- CN110361459A CN110361459A CN201910419253.5A CN201910419253A CN110361459A CN 110361459 A CN110361459 A CN 110361459A CN 201910419253 A CN201910419253 A CN 201910419253A CN 110361459 A CN110361459 A CN 110361459A
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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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
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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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Abstract
The present invention discloses a kind of method for building up of loguat leaf standard decoction finger-print, the method for building up of finger-print, comprising the following steps: 1) preparation of test solution；2) chromatographic condition and system suitability；3) test solution is injected into liquid chromatograph, measures, obtains finger-print.By the research of the ultra performance liquid chromatography finger-print to loguat leaf standard decoction, a kind of preferable loguat leaf standard decoction finger-print has been obtained, has provided new technical method for control said preparation quality.
Description
Technical field
The present invention relates to Pharmaceutical Analysis fields, relate generally to a kind of method for building up of loguat leaf standard decoction finger-print.
Background technique
Standard decocting liquid is standardized to instruct, based on clinical application according to clinical decoction decocting method by theory of traditional Chinese medical science
Single medicinal material medicine materical crude slice decocting liquid made of decoction.Its preparation flow have the characteristics that standardization and it is standardized, can guarantee technique
Unification, and ensure the unification and stabilization of its quality；Standard decocting liquid can be used as a kind of chemical benchmark, for the clinical use of markization
Medicine ensures the accuracy of medication and the consistency of dosage；Also the positive control drug that can be used as drug effect benchmark can be used for evaluating difference
Medicine materical crude slice administration form, to solve the difficult situation of " not homogeneity " caused by due to preparation method difference, to force different medicine materical crude slice to use
Medicine form reaches the effect of " shape is different, and matter is identical ", realizes the consistency of clinical efficacy.
After traditional Chinese medicine fingerprint refers to that certain Chinese medicines or Chinese materia medica preparation are appropriately processed, using certain analysis means,
What is obtained can indicate the chromatogram or spectrogram of its chemical feature.It can more fully reflect chemical component contained by medicinal material
Type and quantity, effectively embody traditional Chinese medicine ingredients globality and comprehensive function, it is quick with its, accurate the features such as transported extensively
For Analysis of Chinese Traditional Medicine identification and quality control etc..
Loguat leaf (scientific name: Eriobotrya japonica Thunb.) is the leaf of rosaceous plant loquat, You Mingba
Leaf, Lu Juye (" Chinese medicine handbook "), former plant loquat also known as: Lu's tangerine (Guangdong) has clearing lung and relieving cough, and stomach function regulating diuresis is quenched the thirst
Effect has treatment lung feverish cough, hemoptysis, bleeding from five sense organs or subcutaneous tissue, stomach hot vomiting effect.But in loguat leaf medicinal material analysis technical field, also still
Not useful ultra performance liquid chromatography method (UPLC) carries out Quality Control Analysis to loguat leaf standard decoction and reports its fingerprint
Map.
Therefore, the existing technology needs to be improved and developed.
Summary of the invention
In view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of loguat leaf standard decoction finger-prints
Method for building up has been obtained a kind of preferable by the research of the ultra performance liquid chromatography finger-print to loguat leaf standard decoction
Loguat leaf standard decoction finger-print provides new technical method for control said preparation quality, it is intended to solve at present to loquat
The problem of the Quality Control Technology deficiency of leaf standard decoction.
Technical scheme is as follows:
A kind of method for building up of loguat leaf standard decoction finger-print, wherein the following steps are included:
1) preparation of test solution: taking loguat leaf standard decoction, adds water, carries out ultrasound or refluxing extraction, lets cool, use water
The weight of less loss is supplied, is centrifuged, filtering；
2) chromatographic condition and system suitability: being stream with acetonitrile using octadecylsilane chemically bonded silica as filler
Dynamic phase A, using 0.05~0.2% acetic acid aqueous solution as Mobile phase B；Column temperature is 25~35 DEG C；Flow velocity is 0.4ml/min, detects wave
A length of 230nm-360nm；The sample volume 1-2 μ l of test solution；
Wherein, gradient elution program is as follows: 0~2min, Mobile phase B: 95%；2~5min, Mobile phase B: 95%；5~
13min, Mobile phase B: 91%；13~25min, Mobile phase B: 78%；25~39min, Mobile phase B: 20%；39~39.1min,
Mobile phase B: 95%；39.1~40min, Mobile phase B: 95%；
3) test solution is injected into liquid chromatograph, measures, obtains finger-print.
Preferably, in step 1), every 0.2g loguat leaf standard decoction, adding concentration is 50%~100% methanol 10ml, ultrasound
It extracts 15~40 minutes；Power is 400W, frequency 40kHz during the ultrasonic extraction.
Preferably, in step 1), add concentration for 70% methanol, ultrasonic extraction 30 minutes.
Preferably, in step 2), using Waters Cortecs T3 chromatographic column, packing material size is 1.6 μm, internal diameter 2.1,
Column length 100mm.
Preferably, in step 2), using 0.1% acetic acid aqueous solution as Mobile phase B；Column temperature is 30 DEG C；Detection wavelength is
326nm。
Preferably, the sample volume of test solution is 1 μ l in step 3).
Preferably, further comprising the steps of:
4) obtained finger-print is subjected to similarity calculation with map is compareed；The control map is as shown in figure 20.
The utility model has the advantages that establishing loguat leaf standard soup using similarity evaluation in the present invention
The UPLC finger-print of agent can be used for the quality control of loguat leaf standard decoction.The fingerprint image of loguat leaf standard decoction of the present invention
Compose construction method, have the advantages that following significant: (1) present invention carries out loguat leaf standard decoction for the first time using UPLC technology
Finger-print research provides new analysis method for the control of loguat leaf standard decoction quality.(2) optimize the building of finger-print
Method, the finger-print baseline made is steady, separating degree is good, peak type is preferable, appearance amount is more.(3) test sample system of the present invention
Make simply, chromatographic condition is easy to accomplish, and precision is high, and its stability and reproducibility are preferable, the detection wave that the present invention uses
Long appearance is more, and the information of reflection is complete.(4) the on-line analysis time is short, saves material cost and time cost.
Detailed description of the invention
Fig. 1 is the top view of full wavelength scanner in the embodiment of the present invention.
Fig. 2 is the 3D view of full wavelength scanner in the embodiment of the present invention.
Fig. 3 is chromatogram when chromatographic column is Waters CORTECS UPLC T3 in the embodiment of the present invention.
Fig. 4 is chromatogram when chromatographic column is Waters UPLC BEHC18 in the embodiment of the present invention.
Fig. 5 is when chromatographic column is Thermo AcclaimTM RSLC PolarAdvantage II in the embodiment of the present invention
Chromatogram.
Fig. 6 is chromatogram when water phase is water in the embodiment of the present invention.
Fig. 7 is chromatogram when water phase is 0.1% formic acid in the embodiment of the present invention.
Fig. 8 is chromatogram when water phase is 0.1% acetic acid in the embodiment of the present invention.
Fig. 9 is chromatogram when Mobile phase B is 0.05% acetic acid in the embodiment of the present invention.
Figure 10 is chromatogram when Mobile phase B is 0.1% acetic acid in the embodiment of the present invention.
Figure 11 is chromatogram when Mobile phase B is 0.2% acetic acid in the embodiment of the present invention.
Figure 12 is chromatogram when column temperature is 25 DEG C in the embodiment of the present invention.
Figure 13 is chromatogram when column temperature is 30 DEG C in the embodiment of the present invention.
Figure 14 is chromatogram when column temperature is 35 DEG C in the embodiment of the present invention.
Figure 15 is the chromatogram that specificity tests negative control solution in the embodiment of the present invention.
Figure 16 is the chromatogram of specificity experiment contrast product solution in the embodiment of the present invention.
Figure 17 is the chromatogram that specificity tests test solution in the embodiment of the present invention.
Figure 18 is the finger-print comparison diagram of different column temperatures in the embodiment of the present invention.
Figure 19 is 15 batches of different batches loguat leaf mark soup finger-prints in the embodiment of the present invention.
Figure 20 is the control map generated in the embodiment of the present invention.
Specific embodiment
The present invention provides that a kind of loguat leaf standard decoction finger-print, characteristic spectrum is established and content assaying method, to make
The purpose of the present invention, technical solution and effect are clearer, clear, and the present invention is described in more detail below.It should be appreciated that
The specific embodiments described herein are merely illustrative of the present invention, is not intended to limit the present invention.
Below by way of specific embodiment, the present invention will be further described.
The preparation of loguat leaf standard decoction:
Loguat leaf medicine materical crude slice 200g is taken, it is weighed, 2400mL water is added and impregnates 30min, using health ceramic pot, first intense fire
(500w) boils, then mild fire (200w) decocts 30min, and 400 mesh standard sieves is selected to filter while hot；The dregs of a decoction add 2000mL water again, military
After fire boils, then it is cooked by slow fire 25min, 400 mesh standard sieves filter while hot, merge filtrate twice, are concentrated under reduced pressure at 62 DEG C；Filtrate
Being concentrated under reduced pressure into solid content is 10%~15%, is placed in freeze drier, is freeze-dried to get loguat leaf standard decoction.
Embodiment 1: finger-print is established
One, the investigation of loguat leaf standard decoction chromatographic condition
1, chromatographic condition and system suitability
The chromatographic condition drafted:: using octadecylsilane chemically bonded silica as filler (2.1 × 100mm, 1.6 μm), flowing
Phase A is acetonitrile, and Mobile phase B is 0.1% glacial acetic acid aqueous solution, carries out linear gradient elution by 1 program of table；Flow velocity 0.4ml/min,
30 DEG C of column temperature；Detection wavelength 326nm, sample volume are 1-2 μ l (sample volume is all made of 1 μ l in the present embodiment).Number of theoretical plate is with green
Ortho acid meter is not less than 5000.
1 eluent gradient table of table
| Time (min) | Flow velocity (ml/min) | Mobile phase B (%) |
| 0 | 0.4 | 95 |
| 2 | 0.4 | 95 |
| 5 | 0.4 | 91 |
| 13 | 0.4 | 90 |
| 25 | 0.4 | 78 |
| 39 | 0.4 | 20 |
| 39.1 | 0.4 | 95 |
| 40 | 0.4 | 95 |
(1) selection of Detection wavelength
Experimentation: on the basis of the experiment condition drafted above, 190 are carried out to test solution using PDA detector
The scanning of~600nm wave band, and the chromatogram of test solution is extracted, as depicted in figs. 1 and 2.
The result shows that: the UV absorption of the chromatographic peak of loguat leaf mark soup concentrates near 230nm-360nm, and 230nm
~300nm, unstability of base line, impurity interference is big, therefore selects 326nm as loguat leaf standard decoction characteristic spectrum in the present invention
Detection wavelength.
(2) selection of chromatographic column
Under identical gradient, investigating chromatographic column respectively is Waters CORTECS UPLC T3 (2.1 × 100mm, 1.6 μ
M), Waters UPLC BEHC18 (2.1 × 100mm, 1.7 μm), Thermo AcclaimTM RSLC PolarAdvantage
When II (PA2) (2.1 × 100mm, 2.2 μm), the influence to chromatogram.As a result as seen in figures 3-5.
The result shows that: when using Waters CORTECS T3 column, chromatographic peak whole separation effect is best, therefore, the present invention
Middle selection Waters CORTECS T3 (Waters Cortecs T3,1.6 μm, 2.1*100mm) is loguat leaf characteristic spectrum
Liquid-phase chromatographic column.
(3) selection of mobile phase water phase acid
When investigation water phase is water, 0.1% formic acid, 0.1% acetic acid respectively, the influence to chromatographic peak.As a result such as the institute of Fig. 6~8
Show.
The result shows that: when Mobile phase B is water, 2.2 minutes and two peaks disappearances in 2.8 minutes；When Mobile phase B is 0.1%
When formic acid, peak is more miscellaneous within 4-6 minutes；When Mobile phase B is 0.1% acetic acid, preferable peak is responded for two separation within 4-6 minutes, and
And entirety baseline is more steady.Therefore, selective flow water phase is 0.1% glacial acetic acid aqueous solution.
(4) selection of acetic acid concentration
Investigating Mobile phase B is respectively 0.05% acetic acid, 0.1% acetic acid, 0.2% acetic acid, and other conditions are same as above, measurement result
As shown in Fig. 9~Figure 11.
The results show that Mobile phase B acetic acid concentration is 0.05%~0.2% little on map influence.This research Mobile phase B is
0.1% acetic acid.
(5) selection of column temperature
When investigation column temperature is 25 DEG C, 30 DEG C, 35 DEG C respectively, the influence to chromatogram peak out.
It can be seen that from Figure 12~Figure 14, for column temperature at 25~30 DEG C, chromatographic peak can reach preferable separation.Column temperature is 25
At~30 DEG C, measurement result is influenced little.This research uses column temperature for 30 DEG C.
In conclusion loguat leaf standard decoction finger-print chromatographic condition determines are as follows: chromatographic column Waters Cortecs T3
(2.1 × 100mm, 1.6 μm), mobile phase A are acetonitrile, and Mobile phase B is 0.1% glacial acetic acid aqueous solution, are carried out by 1 program of table linear
Gradient elution；Flow velocity 0.4ml/min, 30 DEG C of column temperature；Detection wavelength 326nm.
Two, the foundation of sample solution preparation method
1, extracting mode selects
4 parts, every part of 0.2g of loguat leaf standard decocting liquid are weighed, is divided into two groups of A, B, respectively accurate plus methanol 10mL, weighing, A
Group ultrasound 30 minutes, B group water-bath reflux 30 minutes.It is cooled to room temperature, weighs, the weight of less loss is supplied with methanol.Filtering, takes continuous
Filtrate, measurement.As a result as shown in table 2 below.Power is 400W, frequency 40kHz during the ultrasonic extraction.
2 extracting mode of table investigates result
The result shows that: ultrasonic extraction is good compared with refluxing extraction effect.Therefore selection ultrasonic extraction.
2, Extraction solvent selects
3 parts, every part of 0.2g of loguat leaf standard decocting liquid are weighed, respectively accurate plus methanol, ethyl alcohol, each 10mL of water, weighing, ultrasound
30 minutes.It puts to room temperature, the weight of less loss is supplied in weighing with coordinative solvent.Filtering, takes subsequent filtrate, measures.As a result such as the following table 3
It is shown.
Influence of 3 Extraction solvent of table to peak area
| Solvent | Sample weighting amount (g) | Total peak area (mAu*min) | Peak area/sample weighting amount |
| Methanol | 0.2002 | 55.9460 | 279.45 |
| Ethyl alcohol | 0.2006 | 30.2430 | 150.76 |
| Water | 0.2020 | 56.2032 | 278.23 |
The result shows that: when Extraction solvent is ethyl alcohol, chromatographic peak is less；When Extraction solvent is first alcohol and water, chromatographic peak quantity
Response is not much different.When water extracts, big polarity chromatographic peak is higher；When methanol extracts, low polarity chromatographic peak is higher, to guarantee with after
Continuous medicinal material research is corresponding, is preferably extracted using methanol in the present invention program.
3, Extraction solvent concentration is investigated
3 parts, every part of 0.2g of loguat leaf standard decocting liquid are weighed, accurate plus methanol, 50% methanol, 70% methanol are each respectively
10mL, weighing, ultrasound 30 minutes.It puts to room temperature, the weight of less loss is supplied in weighing with coordinative solvent.Filtration, takes subsequent filtrate, surveys
It is fixed.As a result as shown in table 4 below.
The investigation result of 4 Extraction solvent concentration of table
The result shows that: various concentration methanol extraction effect difference is little.But when 50% methanol extracts, the more difficult filtration of solution；
Methanol Extraction solvent effect is higher, therefore selects 70% methanol ultrasonic extraction.
4, extraction time is investigated
3 parts, every part of 0.2g of loguat leaf standard decocting liquid are weighed, respectively accurate plus 70% methanol 10mL, weighing, difference ultrasound 15
Minute, 30 minutes and 45 minutes.It puts to room temperature, the weight of less loss is supplied in weighing with coordinative solvent.Filtering, takes subsequent filtrate, surveys
It is fixed.As a result as shown in table 5 below.
Influence of 5 ultrasonic time of table to peak area
| Time (min) | Sample weighting amount (g) | Total peak area (mAu*min) | Peak area/sample weighting amount |
| 15 | 0.2022 | 20.3438 | 100.61 |
| 30 | 0.2003 | 20.5994 | 102.84 |
| 45 | 0.2012 | 20.0104 | 99.46 |
The result shows that: 30 minutes extraction effects are best, and therefore, final choice extraction time is 30 minutes.
In conclusion the preparation method of loguat leaf standard decoction finger-print test solution determines are as follows: take loguat leaf mark
Soup 0.2g, it is accurately weighed, it sets in stuffed conical flask, precision is added 70% methanol 10ml, weighed weight, it is let cool after ultrasonic 30min,
Weighing, the amount of less loss is supplied with 70% methanol, is shaken up, filter, take subsequent filtrate to get.
Three, methodology validation
1, specificity is tested
Precision draws negative control solution (70% methanol), reference substance solution (chlorogenic acid, 97.314 μ g/ml), test sample
Each 1 μ l of solution is measured by sample introduction under set chromatographic condition.As seen in figs. 15-17, negative control solution should be without corresponding color
Spectral peak.Illustrate that test solution determining fingerprint pattern is noiseless, method is exclusive good.
The result shows that negative control solution is without corresponding chromatographic peak, it was demonstrated that method specificity is good.
2, precision test
(1) instrument precision
1 part of test solution (B10) is taken, is pressed " 4.2.1.6 " item chromatographic condition METHOD FOR CONTINUOUS DETERMINATION 6 times.Record finger-print, band
Enter similarity software, each finger-print there should be 11 characteristic peaks, and calculate separately the similarity with reference fingerprint.It is tied
Fruit is as shown in table 6 below.
6 precision experiment result of table
| Serial number | 1 | 2 | 3 | 4 | 5 | 6 | Mean value | RSD |
| Similarity | 0.989 | 0.989 | 0.987 | 0.987 | 0.988 | 0.987 | 0.988 | 0.10% |
The results show that in 6 measurement results, each finger-print and reference fingerprint similarity are all larger than 0.98, and
RSD is less than 3%.Show that the precision of method is good.
(2) repeated
Laboratory technician's first prepares 6 parts of test solution of loguat leaf standard decocting liquid (B10) finger-print as required,
By being measured under set chromatographic condition on ThermoUHPLC.Finger-print is recorded, brings similarity software into, each finger-print is answered
There are 11 characteristic peaks, and calculates separately the similarity with reference fingerprint.Its result is as shown in table 7 below.
7 repeated experiment result of table
The results show that in 6 measurement results, each finger-print and reference fingerprint similarity are all larger than 0.98, and
RSD is less than 3%.Show that the repeatability of method is good.
(3) Intermediate precision
Laboratory technician's second prepares 6 parts of test solution of loguat leaf standard decocting liquid (B10) finger-print as required, in Water
By being measured under set chromatographic condition on ACQUITY H-class UPLC.Finger-print is recorded, brings similarity software, each finger into
Line map should have 11 characteristic peaks, and calculate separately the similarity with reference fingerprint.Its result is as shown in table 8 below.
8 finger-print Intermediate precision experimental result of table
| Serial number | 1 | 2 | 3 | 4 | 5 | 6 | Mean value | RSD | With repeated RSD |
| Similarity | 0.991 | 0.995 | 0.991 | 0.993 | 0.993 | 0.992 | 0.993 | 0.15% | 0.44% |
The results show that measuring each finger-print in the case where different personnel prepare and being all larger than with reference fingerprint similarity
0.98, RSD less than 3%, calculates RSD less than 3% with repeatability., show that tightness is good among this method.
(4) durability
(a) study on the stability
On the basis of the experiment condition drafted above, 1 part of test solution is taken, it was surveyed respectively at 0,2,4,8,16,24 hour
It is fixed.Finger-print is recorded, brings similarity software into, each finger-print there should be 11 characteristic peaks, calculate separately each period
Finger-print and 0 hour fingerprint similarity.As a result as shown in table 9 below.
9 finger-print stability experiment relative retention time result of table
| Time (h) | 0 | 2 | 4 | 8 | 16 | 24 | Mean value | RSD |
| Similarity | 1.000 | 0.999 | 0.999 | 0.999 | 1.000 | 0.999 | 0.999 | 0.05% |
The results show that in different time sections measurement result, each finger-print and 0 hour fingerprint similarity are big
In 0.99, and RSD is respectively less than 3%.Show that sample measurement result in 24 hours is stablized.
(b) column temperature
On the basis of the experiment condition drafted above, 1 part of test solution is taken, sample introduction under the conditions of different column temperatures records
Finger-print calculates similarity.
As a result: 25 DEG C and 35 DEG C of chromatograms and 30 DEG C of chromatogram similarities are all larger than 0.99, and chromatogram is as shown in figure 18, table
The slight change of bright this method column temperature influences very little to experimental result.
Three, the foundation of map is compareed
The measurement that using the method drafted 15 batches of samples are carried out with single needle finger-print, records finger-print, such as Figure 19 institute
Show, import " similarity evaluation " (2.0 editions) similarity software, uses average value to compare map
Generating mode, Supplements generate control map, and find 11 shared fingerprint peaks, as shown in figure 20.Wherein, peak 1: fresh green
Ortho acid；Peak 3: chlorogenic acid；Peak 4: Cryptochlorogenic acid peak.
Four, it measures
Using similarity evaluation, UPLC figure is generated to 15 batches of loguat leaf standard decoction samples
Spectrum, is matched, matching result is as shown in table 10 with map is compareed.The experimental results showed that 15 batches of loguat leaf standard decoctions (freeze
Dry powder) similarity is above 0.9, each quality of lot stable homogeneous.
The finger-print matching result of 10 15 batches of loguat leaf standard decoctions of table
It should be understood that the application of the present invention is not limited to the above for those of ordinary skills can
With improvement or transformation based on the above description, all these modifications and variations all should belong to the guarantor of appended claims of the present invention
Protect range.
Claims (7)
1. a kind of method for building up of loguat leaf standard decoction finger-print, which comprises the following steps:
1) preparation of test solution: taking loguat leaf standard decoction, adds water, carries out ultrasound or refluxing extraction, is centrifuged, filtering；
2) chromatographic condition and system suitability: using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase
A, using 0.05~0.2% acetic acid aqueous solution as Mobile phase B；Column temperature is 25~35 DEG C；Flow velocity is 0.4ml/min, and Detection wavelength is
230nm-360nm；The sample volume of test solution is 1-2 μ l；
Wherein, gradient elution program is as follows: 0~2min, Mobile phase B: 95%；2~5min, Mobile phase B: 95%；5~13min,
Mobile phase B: 91%；13~25min, Mobile phase B: 78%；25~39min, Mobile phase B: 20%；39~39.1min, mobile phase
B:95%；39.1~40min, Mobile phase B: 95%；
3) step (1) is prepared into gained test solution and injects liquid chromatograph, be measured based on step (2) condition,
Obtain finger-print.
2. the method for building up of loguat leaf standard decoction finger-print according to claim 1, which is characterized in that step 1)
In, every 0.2g loguat leaf standard decoction adds concentration for 50%~100% methanol 10ml, and ultrasonic extraction 15~40 minutes；It is described super
Power is 400W, frequency 40kHz in sound extraction process.
3. the method for building up of loguat leaf standard decoction finger-print according to claim 3, which is characterized in that step 1)
In, add concentration for 70% methanol, ultrasonic extraction 30 minutes.
4. the method for building up of loguat leaf standard decoction finger-print according to claim 1, which is characterized in that step 2)
In, using Waters Cortecs T3 chromatographic column, packing material size is 1.6 μm, internal diameter 2.1, column length 100mm.
5. the method for building up of loguat leaf standard decoction finger-print according to claim 1, which is characterized in that step 2)
In, using 0.1% acetic acid aqueous solution as Mobile phase B；Column temperature is 30 DEG C；Detection wavelength is 326nm.
6. the method for building up of loguat leaf standard decoction finger-print according to claim 1, which is characterized in that in step 3)
The sample volume of test solution is 1 μ l.
7. the method for building up of loguat leaf standard decoction finger-print according to claim 1, which is characterized in that further include with
Lower step:
4) obtained finger-print is subjected to similarity calculation with map is compareed；The control map is as shown in figure 20.
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| CN115201376A (en) * | 2022-07-18 | 2022-10-18 | 国药集团精方（安徽）药业股份有限公司 | Quality detection method of loquat cough relieving granules |
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