CN107727754A - A kind of HPLC fingerprint atlas detection methods of Xiaojing - Google Patents
A kind of HPLC fingerprint atlas detection methods of Xiaojing Download PDFInfo
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- G—PHYSICS
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- 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
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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Abstract
The invention discloses a kind of traditional Chinese medicine fingerprint detection method of Xiaojing, the present invention to conditions such as instrument, chromatographic column, mobile phase, Detection wavelengths by screening, establish liquid-phase fingerprint condition determination and carried out methodological study, Xiaojing finger-print standard has been formulated according to more batches big production sample.Because the product prescription contains the medicine sources such as autonomic drug, animal drugs and resin, crude drug source is extensive, the place of production is numerous, very different, and the not high limitation of quality standard be present, in order to further improve product quality, ensure drug safety, effectively, intend being controlled Xiaojing from medicine elementary source, product quality can fully and effectively be controlled using finger-print, overall process quality of production control system is established, the quality formulated on this basis through full industrial chain can trace system.
Description
Technical field
The invention belongs to field of traditional Chinese medicine detection, it is related to a kind of HPLC fingerprint atlas detection methods of Xiaojing.
Background technology
Chinese medicine fingerprint be the shared similitude using Chinese medicine colony as theoretical foundation, by various spectrum, wave spectrum, chromatogram etc.
A kind of traditional Chinese medicine quality control model that technology grows up.After traditional Chinese medicine fingerprint refers to that Chinese medicine is appropriately processed, using certain
Analysis means, what is obtained can indicate the collection of illustrative plates at the shared peak of the Chinese medicine characteristic, so as to discerning the false from the genuine, evaluate quality of medicinal material
Homogeneity and stability.Within the specific limits, traditional Chinese medicine fingerprint can reflect Chinese medicine overall picture substantially, make its quality control index by original
The measure to single component content rise to detection to whole Chinese medicine interior quality, realize the synthesis to Chinese medicine inherent quality
Evaluation and comprehensive control of overall material, traditional Chinese medicine quality is set to reach stable, controllable, it is ensured that the stabilization of tcm clinical practice curative effect, and make
Traditional Chinese medicine research more meets the organic conception of motherland's medical science.The detection method of traditional Chinese medicine fingerprint has chromatography, spectroscopic methodology, spectral method
Deng.Chromatography mainly includes thin-layer chromatography, high performance liquid chromatography, gas-chromatography, Capillary Electrophoresis etc., wherein high performance liquid chromatography
Method has the characteristics of efficient, quick, sensitive, favorable reproducibility, with UV-detector (UV), diformazan pipe array detector (DAD), steaming
A variety of detector combinations such as Light Scattering Detector (ELSD) and mass detector (MS), available for Various Complex composition in Chinese medicine
Analysis detection, as traditional Chinese medicine fingerprint research main stream approach.
《Chinese Pharmacopoeia》2010 editions one has been recorded a kind of Chinese medicine with the effect of mass dissipating and swelling eliminating, blood-activating analgetic, medicine
Name is " xiaojin pill ", by muscone, semen momordicae, radix aconiti agrestis, resina liquidamberis, frankincense, myrrh, excrementum pteropi, Radix Angelicae Sinensis, earthworm, scented ink etc.
10 taste Chinese medicines are prepared.The party is classical Chinese medicine recipe, is increased available for a variety of difficult and complicated cases such as goitre, benign prostate
The treatment of life, post-herpetic neuralgia, cystic hyperplasia of mammary gland, chronic pelvic inflammatory disease enclosed mass and nodular fasciitis etc..At present
A variety of formulations listings such as existing tablet, capsule, as CN106265992A discloses Xiaojin Capsules and preparation method thereof,
CN102764311A discloses little Jin dispersible tablets and preparation method thereof.Xiaojing is mainly used as medicine using the full powder of raw medicinal herbs at present
Mode.
Because Xiaojing contains the medicine sources such as autonomic drug, animal drugs and resin, crude drug source is extensive, and the place of production is numerous, and the good and the bad is not
Together, and the not high limitation of quality standard be present, in order to further improve product quality, ensure drug safety, effectively, intend to small
Gold preparation is controlled from medicine elementary source, and product quality is fully and effectively controlled using fingerprint pattern technology, establishes overall process life
Yield and quality control system, the quality formulated on this basis through full industrial chain can trace system.
The content of the invention
The quality of Chinese medicine compound prescription can not be controlled comprehensively for current Xiaojing quality determining method, the purpose of the present invention exists
In the HPLC fingerprint atlas detection methods for establishing Xiaojing.
To achieve the above object, the present invention adopts the following technical scheme that：
A kind of HPLC fingerprint atlas detection methods of Xiaojing, the Xiaojing by following parts by weight raw material system
Into：Muscone 1-20 parts, radix aconiti agrestis 40-60 parts, frankincense 20-40 parts, excrementum pteropi 40-60 parts, earthworm 40-60 parts, semen momordicae 40-
60 parts, resina liquidamberis 40-60 parts, myrrh 20-40 parts, Radix Angelicae Sinensis 20-40 parts, scented ink 1-10 parts, the detection method comprises the following steps：
(1) preparation of need testing solution：Xiaojing to be measured is finely ground, 0.5-5 parts by weight are taken, add water saturated positive fourth
Alcohol 10-100 parts by volume, weighed weight, let cool to room temperature after refluxing extraction, then weighed weight, supplied with water saturated n-butanol
The weight of less loss, shakes up, and with filtering with microporous membrane, takes subsequent filtrate, standby；
(2) accurate to draw need testing solution 5-20 μ l, upper high performance liquid chromatograph is detected, the high performance liquid chromatography
Instrument contains UV-detector, and the UV-detector sets Detection wavelength as 200-300nm, the high performance liquid chromatograph
Stationary phase is C18Chromatographic column, mobile phase are the mixed solution for the phosphoric acid solution that methanol is 0.05-0.2% with weight concentration, are used
Gradient elution, flow velocity 0.5-2ml/min, column temperature are 25~40 DEG C, and number of theoretical plate is not less than 10000；
(3) 0~120min of need testing solution chromatogram is recorded, utilizes Chinese Pharmacopoeia Commission's chromatographic fingerprints of Chinese materia medica
Similarity evaluation system, data importing, multiple spot correction and Data Matching are passed through respectively to the chromatogram of need testing solution, produce finger
Line collection of illustrative plates simultaneously carries out similarity analysis,
The relation of the parts by weight and the parts by volume is g/mL.
Preferably, the UV-detector sets Detection wavelength as 254nm.
Preferably, the weight concentration of the phosphoric acid solution is 0.1%.
Preferably, in the gradient elution, the volume ratio difference of 0.1% phosphoric acid solution phase in each period and mobile phase
For：0~2min, phosphoric acid solution phase 70%；2~20min, phosphoric acid solution phase 70~45%；20~50min, phosphoric acid solution phase 45
~25%；50~70min, phosphoric acid solution phase 25~20%；70~85min, phosphoric acid solution phase 20~15%；85~90min, phosphorus
Acid solution phase 15~5%；90~120min, phosphoric acid solution phase 5~0%.
Preferably, there are 20 common characteristic peaks in the chromatogram of the need testing solution, wherein No. 1 peak is that muscone is special
It is the peculiar peak of resina liquidamberis to have peak, No. 4 peaks, No. 8 and No. 14 peaks are the peculiar peak of frankincense, No. 9 peaks be the peculiar peak of excrementum pteropi, No. 15 and 18
Number peak is the peculiar peak of myrrh, using the peak of shared peak 13 of resina liquidamberis and myrrh as reference peak, the relative retention time difference at each peak
It is 5.997min for, No. 1 peak, No. 4 peaks are 18.900min, and No. 8 peaks are 38.318min, and No. 9 peaks are 45.058min, No. 13 peaks
For 59.838min, No. 14 peaks are 69.005min, and No. 15 peaks are 76.934min, and No. 18 peaks are 90.353min.
Involved phosphoric acid solution concentration is volumetric concentration in the present invention, and such as 0.1% phosphoric acid solution refers to often
Contain phosphoric acid 0.1ml in 100ml phosphate aqueous solutions.
The beneficial effects of the invention are as follows：
Because the composition overwhelming majority contained in Chinese medicine can carry out analysis detection on high performance liquid chromatograph, utilize
HPLC methods can make tens kinds of complicated chemical compositions reach good separation.Obtained chromatogram is with the number and peak of chromatographic peak
Area characterizes the number of chemical composition contained by sample and the height of different chemical composition content respectively, so as to reflect Chinese medicine matter
The Global Information of amount；Along with HPLC method separation efficiencies are high, selectivity is high, detection is sensitive, analyze speed is fast, have a wide range of application
Feature, it is a kind of method that most basic, most suitable drafting finger-print, purposes are most wide in all spectrum, chromatography.At this
In invention, establish the HPLC finger-prints of Xiaojing, while determine 10 batches of Xiaojin Capsules, marked 20 shared peaks and
One standard finger-print, resulting chromatogram similarity reach more than 0.97；With its relative retention time and relative peak face
Product comes computational stability, repeatability and precision, and the RSD values of relative retention time are respectively less than 2.0%, the RSD of relative peak area
Value is respectively less than 5.0%；In 20 compositions, identify the peculiar peak of 5 medicinal materials, be respectively muscone, resina liquidamberis, frankincense,
Myrrh, excrementum pteropi.
The Xiaojing finger-print that the present invention establishes can be used for the Ji Yuan of control raw medicinal material, and in process of production may be used
It is effective to instruct feed intake, strictly regulate production operation, really ensure that clinical application safely, effectively, reliably, with other detections
Method is easy to operate, quick compared to having the advantages that.
Brief description of the drawings
Fig. 1 is the HPLC collection of illustrative plates after testing sample methanol ultrasonic extraction.
Fig. 2 is the HPLC collection of illustrative plates after the water saturated n-butanol refluxing extraction of testing sample.
Fig. 3 is the HPLC collection of illustrative plates that 210nm Detection wavelengths obtain.
Fig. 4 is the HPLC collection of illustrative plates that 254nm Detection wavelengths obtain.
Fig. 5 is the HPLC collection of illustrative plates that 267nm Detection wavelengths obtain.
Fig. 6 is the HPLC collection of illustrative plates that 280nm Detection wavelengths obtain.
Fig. 7 is the HPLC collection of illustrative plates that 300nm Detection wavelengths obtain.
Fig. 8 is the HPLC standard finger-prints of Xiaojin Capsules.
Fig. 9 is the HPLC standard finger-prints of Moschus in Xiaojin Capsules prescription.
Figure 10 is the HPLC standard finger-prints of frankincense in Xiaojin Capsules prescription.
Figure 11 is the HPLC standard finger-prints of myrrh in Xiaojin Capsules prescription.
Figure 12 is the HPLC standard finger-prints of resina liquidamberis in Xiaojin Capsules prescription.
Figure 13 is the HPLC standard finger-prints of excrementum pteropi in Xiaojin Capsules prescription.
Figure 14 is the HPLC standard finger-prints of Radix Angelicae Sinensis in Xiaojin Capsules prescription.
Embodiment
Below by taking Xiaojin Capsules as an example, the present invention is described in detail by specific embodiment.
Used Xiaojin Capsules are provided by Jian Min medicine companies Group Plc in following examples.Muscone
30g, semen momordicae 150g, radix aconiti agrestis 150g, resina liquidamberis 150g, frankincense 75g, myrrh 75g, excrementum pteropi 150g, Radix Angelicae Sinensis 75g, earthworm
150g, scented ink 12g, are filled after crushing with capsule.
1. instrument and reagent
Waters e2695 high performance liquid chromatographs, quaternary pump, online vacuum degassing system, automatic sampler, column oven,
UV-detector；AB204-E electronic balances (Shanghai plum Teller-support benefit Instrument Ltd.).
The Xiaojin Capsules (170116-0121,170332-0335, totally 10 crowdes) of 10 batches；Methanol is chromatographic grade, and water is
Purified water.
2. method and step
(1) prepared by need testing solution：Xiaojin Capsules content is taken, it is finely ground, 1g is taken, accurately weighed, precision adds water saturation
N-butanol 50ml, weighed weight, let cool to room temperature after refluxing extraction, then weighed weight, supplied and subtracted with water saturated n-butanol
The weight of mistake, shakes up, and with filtering with microporous membrane, takes subsequent filtrate, standby；
(2) chromatographic condition：Agilent ZORBAX SB C18, 4.6 × 250mm, 5 μm；Mobile phase：The phosphorus of methanol -0.1% is molten
Liquid is mobile phase, gradient elution：A phases are methanol, and B phases are 0.1% phosphoric acid solution；0~2min, B phase 70%；2~20min, B phase
70~45%；20~50min, B phase 45~25%；50~70min, B phase 25~20%；70~85min, B phase 20~15%；85
~90min, B phase 15~5%；90~120min, B phase 5~0%；Flow velocity：1ml/min；Detection wavelength is 254nm；Column temperature 35
℃；The μ l of sample size 10.Number of theoretical plate is pressed No. 13 peaks and calculated, and should be not less than 10000.
(3) methodological study：Precision test, stability test, replica test are carried out according to the requirement of finger-print；
(4) sample determines：Access batch Xiaojin Capsules, prepare 10 parts of need testing solutions, respectively respectively according to step (1) method
Precision draws 10 μ l, is measured using the chromatographic condition of step (2), record 0-120min chromatograms, with No. 13 in chromatogram
Peak is to calculate relative retention time and relative peak area with reference to peak；
(5) foundation of finger-print and peak attribution analysis is carried out：Access batch Xiaojin Capsules sample respectively, by step (1)
Need testing solution is prepared under, is measured by the chromatographic condition of step (2) and records 120min chromatograms, as a result all chromatograms
Peak is concentrated in 120min, compares the chromatogram of batch sample, it is determined that 20 shared peaks, and in finished product chromatographic fingerprinting bar
Under part, finger-print research is carried out to every taste medicinal material in prescription, carries out peak attribution analysis；By with raw medicinal material figure in prescription
Spectrum contrast, confirm No. 1 peak be the peculiar peak of muscone, No. 4 peaks are the peculiar peak of resina liquidamberis, No. 8 and No. 14 peaks be the peculiar peak of frankincense,
No. 9 peaks are the peculiar peak of excrementum pteropi, No. 15 and No. 18 peaks are the peculiar peak of myrrh.Select the peak of shared peak 13 of resina liquidamberis and myrrh
For reference peak, the relative retention time at each peak is respectively that No. 1 peak is 5.997min, and No. 4 peaks are 18.900min, and No. 8 peaks are
38.318min, No. 9 peaks are 45.058min, and No. 13 peaks are 59.838min, and No. 14 peaks are 69.005min, and No. 15 peaks are
76.934min, No. 18 peaks are 90.353min.
(6) sample Similarity Measure：10 batch Xiaojin Capsules chromatograms of gained are imported what NF committee member promulgated
2004 editions " similarity evaluations ", it is 0.1min to choose " time window " width, is generated using sample 1
Control spectrum, chromatographic fingerprinting common pattern is generated by multiple spot correction and Data Matching, the finger-print of 10 batches of samples is entered
Row similarity analysis.
3. prepared by need testing solution：According to the material composition of Xiaojin Capsules, petroleum ether, methanol, 95% second have been investigated respectively
Alcohol, water saturated n-butanol are as Extraction solvent, by comprehensive analysis, water saturated n-butanol extraction gained need testing solution energy
Main component in enough more comprehensively reflection finished products, and repeatability is preferably, so determining that water saturated n-butanol is molten as extracting
Agent；Extracting method (ultrasound, backflow) is investigated again, it is found that the test sample finger-print baseline of refluxing extraction carries than ultrasound
Make even steady, it is final to determine that preparation finger measure test sample preparation method is：Take Xiaojin Capsules finely ground, take about 1g, precision claims
Determine, the water saturated n-butanol 50ml of precision addition, weighed weight, let cool after refluxing extraction to room temperature, then weighed weight, satisfied with water
The n-butanol of sum supplies the weight of less loss, shakes up, and is filtered with miillpore filter, takes subsequent filtrate, standby.Partial fingerprints collection of illustrative plates see Fig. 1-
2。
4. the foundation of detection method
4.1 instruments, reagent and chromatographic condition
Instrument：Waters e2695 high performance liquid chromatographs, quaternary pump, online vacuum degassing system, automatic sampler, post
Incubator,
Reagent：Methanol is chromatographically pure, and n-butanol is pure to analyze, and water is purified water.
Chromatographic column：Agilent ZORBAX SB-C18, 4.6 × 250mm, 5 μm；Detection wavelength：210,254,267,280,
300nm；The μ l of sample size 10；Column temperature：30~40 DEG C；Flow velocity：1ml/min.
The selection of 4.2 mobile phases
Through comparing, screening, find the phosphoric acid solution systems of (A) methanol-(B) 0.1% with linear gradient elution, gained liquid phase color
Spectrogram baseline compared with steady, chromatographic peak peak shape compared with it is symmetrical, separating degree is preferable.
The optimization of 4.3 gradient elution programs：Grope through system, optimize, it is found that following linear gradient elution program can be compared with
Chemical composition in comprehensive detection preparation, can make these compositions obtain comparatively ideal separating effect again：0~2min, B phase
70%；2~20min, B phase 70~45%；20~50min, B phase 45~25%；50~70min, B phase 25~20%；70~
85min, B phase 20~15%；85~90min, B phase 15~5%；90~120min, B phase 5~0%；Complete 1 sample analysis
Program.The record time is 0~120min.
The determination of 4.4 optimum determining wavelength：Comparative analysis through liquid chromatogram collection of illustrative plates, discovery 210,267,280,300nm
Preparation collection of illustrative plates in baseline it is relatively flat, peak shape is pretty good, but appearance is less, it is impossible to the composition of reflection preparation comprehensively；Under 254nm, respectively
Composition fingerprint peakses separating degree is preferable, considers number, baseline and the signal response power of chromatographic peak, it is fingerprint to determine 254nm
The best detection wavelength of collection of illustrative plates.Finger-print is shown in Fig. 3-7.
5. methodological study
5.1 stability test：Need testing solution is taken to determine, investigate each in 0,4,8,12h according to condition under above-mentioned chromatogram item
The relative retention time at shared peak and the RSD of relative peak area.As a result show, the relative retention time of each chromatographic peak is respectively less than
1.0%, the relative peak area of each chromatographic peak is respectively less than 3.0%, meets fingerprint pattern technology requirement.It is shown in Table 1-2.
The study on the stability relative retention time of table 1
The study on the stability relative peak area of table 2
5.2 replica test：Same 6 parts of sample lots are taken, 6 parts of confessions are prepared according to 1 lower need testing solution preparation method
Test sample solution, it is measured under above-mentioned chromatographic condition, investigates the relative retention time and relative peak area at each shared peak
RSD.As a result show, the relative retention time of each chromatographic peak is respectively less than 1.0%, and the relative peak area of each chromatographic peak is respectively less than
4.0%, meet fingerprint pattern technology requirement.It is shown in Table 3-4.
The repeatability of table 3 investigates relative retention time
The repeatability of table 4 investigates relative peak area
5.3 precision test：Need testing solution is taken continuous sample introduction 6 times, to be investigated each shared according to condition under above-mentioned chromatogram item
The relative retention time at peak and the RSD of relative peak area.As a result showing, the relative retention time of each chromatographic peak is respectively less than 1.0%,
The relative peak area of each chromatographic peak is respectively less than 4.0%, meets fingerprint pattern technology requirement.It is shown in Table 5-6.
The precision of table 5 investigates relative retention time
The precision of table 6 investigates relative peak area
6. the foundation of finger-print：10 batches of Xiaojin Capsules samples are taken respectively, prepare need testing solution, are determined and are recorded
125min chromatograms, as a result all chromatographic peaks concentrate in 125min, compare the chromatogram of 10 batches of samples, it is determined that 20 altogether
There is peak.By confirming, selection retention time is reference peak for 59.838min No. 13 peaks；
6.1 sample Similarity Measures：10 batches of Xiaojin Capsules chromatograms are imported to " the Chinese medicine color of Chinese Pharmacopoeia Commission's promulgation
Compose fingerprint similarity evaluation system (2004A versions) ", it is 0.1min to choose " time window " width, is generated and compareed using sample 1
Spectrum, chromatographic fingerprinting common pattern is generated by multiple spot correction and Data Matching, 10 batches of sample fingerprint similarities calculate
It the results are shown in Table 7.
70 batches of sample similarities of table
| Lot number | Similarity |
| 170116 | 0.987 |
| 170117 | 0.994 |
| 170118 | 0.983 |
| 170119 | 0.993 |
| 170120 | 0.994 |
| 170121 | 0.981 |
| 170332 | 0.996 |
| 170333 | 0.991 |
| 170334 | 0.977 |
| 170335 | 0.981 |
| Reference fingerprint | 1 |
The formulation of 6.2 Xiaojin Capsules standard finger-prints
More batches of Xiaojin Capsules are taken, need testing solution are made by the preparation method of the need testing solution respectively, according to detection
More batches of detection data of gained, establish the HPLC-UV detection of Xiaojin Capsules preparation, establish high-efficient liquid phase color spectral peak retention time
And peak area value, all detection data are then collected, calculate the average value and relative peak area of each corresponding peak relative retention time
Average value and standard deviation etc., be divided by respectively with the Average residence time with reference to peak and average peak area, as standard fingerprint
Relative the peak retention time and relative peak area value of collection of illustrative plates, result of calculation is exported, formulate standard finger-print data, draw standard
Finger-print, standard finger-print are shown in Fig. 8, the results are shown in Table 8.
The standard finger-print data of table 8
Correlation between 6.3 medicinal materials and finished product finger-print
The preparation of medicinal material need testing solution：Take ingredients in prescription that glue is made according to Xiaojin Capsules preparation process respectively
Capsule, synergy is carried out according to standard finger-print condition.Finger-print is shown in Fig. 9-14, the results are shown in Table 9.
The Xiaojin Capsules of table 9 and medicinal material correlation collection of illustrative plates
Claims (5)
1. a kind of HPLC fingerprint atlas detection methods of Xiaojing, the Xiaojing by following parts by weight raw material system
Into：Muscone 1-20 parts, radix aconiti agrestis 40-60 parts, frankincense 20-40 parts, excrementum pteropi 40-60 parts, earthworm 40-60 parts, semen momordicae 40-
60 parts, resina liquidamberis 40-60 parts, myrrh 20-40 parts, Radix Angelicae Sinensis 20-40 parts, scented ink 1-10 parts, it is characterised in that the detection method bag
Include following steps：
(1) preparation of need testing solution：Xiaojing to be measured is finely ground, 0.5-5 parts by weight are taken, add water saturated n-butanol
10-100 parts by volume, weighed weight, let cool to room temperature after refluxing extraction, then weighed weight, supplied and subtracted with water saturated n-butanol
The weight of mistake, shakes up, and with filtering with microporous membrane, takes subsequent filtrate, standby；
(2) accurate to draw need testing solution 5-20 μ l, upper high performance liquid chromatograph is detected, and the high performance liquid chromatograph contains
There is UV-detector, the UV-detector sets Detection wavelength as 200-300nm, the fixation of the high performance liquid chromatograph
It is mutually C18Chromatographic column, mobile phase are the mixed solution of methanol and 0.05-0.2% phosphoric acid solutions, and using gradient elution, flow velocity is
0.5-2ml/min, column temperature are 25~40 DEG C, and number of theoretical plate is not less than 10000；
(3) 0~120min of need testing solution chromatogram is recorded, it is similar using Chinese Pharmacopoeia Commission's chromatographic fingerprints of Chinese materia medica
Evaluation system is spent, data importing, multiple spot correction and Data Matching are passed through respectively to the chromatogram of need testing solution, produce fingerprint image
Compose and carry out similarity analysis,
The relation of the parts by weight and the parts by volume is g/mL.
2. the HPLC fingerprint atlas detection methods of Xiaojing as claimed in claim 1, it is characterised in that：The UV-detector
Set Detection wavelength as 254nm.
3. the HPLC fingerprint atlas detection methods of Xiaojing as claimed in claim 1, it is characterised in that：The phosphoric acid solution
Concentration is 0.1%.
4. the HPLC fingerprint atlas detection methods of Xiaojing as claimed in claim 3, it is characterised in that：The gradient elution
In, the volume ratio of 0.1% phosphoric acid solution phase is respectively in each period and mobile phase：0~2min, phosphoric acid solution phase 70%；2~
20min, phosphoric acid solution phase 70~45%；20~50min, phosphoric acid solution phase 45~25%；50~70min, phosphoric acid solution phase 25
~20%；70~85min, phosphoric acid solution phase 20~15%；85~90min, phosphoric acid solution phase 15~5%；90~120min, phosphorus
Acid solution phase 5~0%.
5. the HPLC fingerprint atlas detection methods of Xiaojing as claimed in claim 1, it is characterised in that：The need testing solution
Chromatogram in have 20 common characteristic peaks, wherein No. 1 peak is the peculiar peak of muscone, No. 4 peaks be the peculiar peak of resina liquidamberis, No. 8
Be the peculiar peak of frankincense with No. 14 peaks, No. 9 peaks are the peculiar peak of excrementum pteropi, No. 15 and No. 18 peaks are the peculiar peak of myrrh, with resina liquidamberis and
The peak of shared peak 13 of myrrh is reference peak, and the relative retention time at each peak is respectively that No. 1 peak is 5.997min, and No. 4 peaks are
18.900min, No. 8 peaks are 38.318min, and No. 9 peaks are 45.058min, and No. 13 peaks are 59.838min, and No. 14 peaks are
69.005min, No. 15 peaks are 76.934min, and No. 18 peaks are 90.353min.
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| CN112285234A (en) * | 2020-10-22 | 2021-01-29 | 健民药业集团股份有限公司 | Gas phase fingerprint spectrum analysis method of traditional Chinese medicine |
| CN112444588A (en) * | 2019-08-27 | 2021-03-05 | 中国科学院大连化学物理研究所 | Fingerprint spectrum detection method for quality of Xiaojin capsules, Xiaojin pills or Xiaojin tablets |
| CN112684073A (en) * | 2021-01-08 | 2021-04-20 | 健民药业集团股份有限公司 | Method for simultaneously detecting contents of two volatile components in metabolism of gold microcapsule prototype |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112444588A (en) * | 2019-08-27 | 2021-03-05 | 中国科学院大连化学物理研究所 | Fingerprint spectrum detection method for quality of Xiaojin capsules, Xiaojin pills or Xiaojin tablets |
| CN110907545A (en) * | 2019-11-14 | 2020-03-24 | 健民药业集团股份有限公司 | Method for simultaneously determining contents of six metabolites in small gold preparation |
| CN112285234A (en) * | 2020-10-22 | 2021-01-29 | 健民药业集团股份有限公司 | Gas phase fingerprint spectrum analysis method of traditional Chinese medicine |
| CN112684073A (en) * | 2021-01-08 | 2021-04-20 | 健民药业集团股份有限公司 | Method for simultaneously detecting contents of two volatile components in metabolism of gold microcapsule prototype |
| CN112684073B (en) * | 2021-01-08 | 2022-07-08 | 健民药业集团股份有限公司 | Method for simultaneously detecting contents of two volatile components in metabolism of gold microcapsule prototype |
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