CN109324141A - The characteristic fingerprint pattern and its construction method of a kind of wide dragon or its processed product and application - Google Patents
The characteristic fingerprint pattern and its construction method of a kind of wide dragon or its processed product and application Download PDFInfo
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Abstract
The construction method of wide dragon of the invention or its processed product characteristic fingerprint pattern is the following steps are included: prepared by (1) test solution: wide dragon health product or its processed product being crushed, NaCl solution, ultrasonic extraction is added, centrifugation, takes supernatant, constant volume, filtering, obtains test solution；(2) prepared by reference substance solution: taking hypoxanthine, inosine, uracil reference substance, adds NaCl solution to dissolve, obtain reference substance solution；(3) high performance liquid chromatography detection constructs wide dragon health product or the characteristic fingerprint pattern of its processed product.The present invention uses hypoxanthine, inosine, uracil, for main Testing index, can effectively detect the miscellaneous peak in elution process as reference substance solution, can accurately be calculated wide dragon health product and processed product；The method of the present invention is easy to operate, stable, reproducible, can carry out more perfect quality evaluation to wide dragon health product and processed product, can rapidly and accurately identify wide dragon health product and processed product.
Description
Technical field
The present invention relates to the characteristic fingerprint pattern and its construction method of a kind of wide dragon or its processed product and application, belong to
Medicine detection technique field.
Background technique
Wide dragon is the hirudo leech of Ju Yin section animal Pheretima aspergillum Pheretima aspergillum (E.Perrier).Tool
Heat-clearing arresting convulsion dredging collateral, is relievingd asthma, the function of diuresis, for unconsciousness due to high fever, twitch, arthralgia, extremity numbness, hemiplegia,
Dyspnea and cough due to lung-heat, edema and little urine.
The concocting method and processing action of Radix Glycyrrhizae pheretima are in " Guangdong Province's prepared slices of Chinese crude drugs concocted specification " version first in 2011
It is on the books in volume, " Guangdong Chinese medicinal herbal " 1994 editions first volumes, but in " Guangdong Province's Chinese medicine standard " and successive dynasties " Chinese Pharmacopoeia "
In recorded without correlation.The characteristic concocting method that wide dragon is Guangdong Lingnan area is processed using Radix Glycyrrhizae, Radix Glycyrrhizae is in tcm prescription
In frequently as efficacy-enhancing ingredient, coordinating the drug actions of a prescription.After Radix Glycyrrhizae processes, can make wide dragon quality is crisp, synergy subtract it is dry, rectify smelly flavoring, with
Convenient for taking.
Wide dragon is processed using Radix Glycyrrhizae and meets traditional Chinese medicine theory, and is widely applied for a long time by In Guangdong Province people.But
It is existing to process the technique of wide dragon still in more coarse situation, the process procedures such as immersion, supplementary product consumption, drying with Radix Glycyrrhizae
Details is indefinite, is unfavorable for the promotion and application of the concocting method.Also without corresponding quality standard, quality is difficult to obtain effectively
Control, hinder the further development of wide dragon Radix Glycyrrhizae preparation method.
The distinctive concocting method in In Guangdong Province is " Radix Glycyrrhizae system ", is recorded in 1984 editions " Guangdong Province's prepared slices of Chinese crude drugs concocted specification "
The method for processing pheretima with Radix Glycyrrhizae, technique are as follows: it takes pure land imperial, puts people temperature Radix Glycyrrhizae bubbly water 2 hours, pick up, dry, dissection,
Weed out grey bits；Every 100kg pure land dragon, with Radix Glycyrrhizae 20kg (cooking).Separately there are other preparation methods also to continue to use in place, such as " national Chinese medicine
Concocted specification " in wine pheretima；Wine moistens sand frying pheretima in " Zhejiang Province's Chinese medicine preparation specification "；" Hunan Province's Chinese medicine preparation rule
Model " in talcum powder pheretima etc..
Chemical component is more complicated in wide dragon, and the assay of single chemical component is difficult to evaluate its quality, and Chinese medicine
Finger-print then can more fully reflect in it quality.Pheretima has the report of the chromatographic fingerprintings such as efficient liquid phase at present
Road, but have no the report for thering is Radix Glycyrrhizae to process wide dragon finger-print both at home and abroad.Therefore, with the side of HPLC traditional Chinese medicine fingerprint
Method establishes the HPLC finger-print of wide dragon processed product and its health product, carries out comprehensive study point to the quality of processed product and health product
Analysis.
Summary of the invention
A kind of wide dragon or its processed product are provided it is an object of the invention to overcome above-mentioned the deficiencies in the prior art place
Characteristic fingerprint pattern and its construction method and application, method of the invention can rapidly and accurately identify wide dragon health product and big gun
Product, this method result is accurate, precision is high.
To achieve the above object, the technical scheme adopted by the invention is as follows: the characteristic fingerprint of a kind of wide dragon or its processed product
The construction method of map, comprising the following steps:
(1) prepared by test solution: wide dragon health product or its processed product are crushed, addition NaCl solution, ultrasonic extraction, from
The heart takes supernatant, and with NaCl solution constant volume, filtering obtains test solution；
(2) prepared by reference substance solution: taking hypoxanthine, inosine, uracil reference substance, NaCl solution is added to dissolve, obtain pair
According to product solution；
(3) high performance liquid chromatography detection constructs wide dragon health product or the characteristic fingerprint pattern of its processed product.
As the preferred embodiment of construction method of the present invention, in the step (1), the concentration of NaCl solution is
0.9%, the time of ultrasonic extraction is 20~30min, centrifugal rotational speed 5000rmin-1, centrifugation time is 10~20min, is supplied
Wide dragon health product or its processed product and the mass volume ratio of NaCl solution are 1:20 in test sample solution.
As the preferred embodiment of construction method of the present invention, in the step (2), the concentration of NaCl solution is
0.9%, hypoxanthic concentration is 80~100 μ g/mL in reference substance solution, and the concentration of inosine is 80~100 μ g/mL, is urinated phonetic
The concentration of pyridine is 80~120 μ g/mL.
As the preferred embodiment of construction method of the present invention, chromatographic condition in the step (3) are as follows:
Chromatographic column: C18 column；
Mobile phase: using 0.01mol potassium dihydrogen phosphate as mobile phase A, ladder is carried out by Mobile phase B of 70% methanol solution
Degree elution, the eluent gradient elution requirement are as follows: 0~5min, A (100%) in mobile phase, B (0%) in mobile phase；5~
20min, A (100% → 85%~80%) in mobile phase, B (0% → 15%~20%) in mobile phase；20~30min, flowing
A (85%~80% → 65%~50%) in phase, B (15%~20% → 35%~50%) in mobile phase；30~40min, stream
Move A (65%~50% → 0%) in phase, B (35%~50% → 100%) in mobile phase；40~45min, A in mobile phase
(0%), B (100%) in mobile phase；
Detector: UV detector；
Flow velocity: 0.8~1.0mL/min；
Detection wavelength: 250~260nm；
Column temperature: 20~25 DEG C；
It is accurate respectively to draw each 10~15 μ L of test solution and control solution, inject high performance liquid chromatograph, measurement.
As the preferred embodiment of construction method of the present invention, building wide dragon or its processing in the step (3)
The method of the characteristic fingerprint pattern of product are as follows: by chromatographic peak appearance time in reference substance solution and test solution chromatogram and
Chromatographic peak obtains the common characteristic peaks in test solution map with the presence or absence of being compared；To 11 batches of wide dragon processed product samples
Product are detected, the wide dragon being made of using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 21 common characteristic peaks
Processed product characteristic fingerprint pattern；11 batches of wide dragon health product samples are detected, are commented using chromatographic fingerprints of Chinese materia medica similarity
The wide dragon health product characteristic fingerprint pattern that valence Software Create is made of 20 common characteristic peaks.
As the preferred embodiment of construction method of the present invention, the wide dragon processed product characteristic fingerprint pattern includes
21 common characteristic peaks, wherein No. 1 peak is uracil, and No. 7 peaks are hypoxanthine, and No. 11 peaks are inosine, other are non-principal component
Peak；
It is referring to peak, the relative retention time of each characteristic peak are as follows: No. 1 peak: 0.356~0.362 with No. 11 peaks of inosine；No. 2
Peak: 0.374~0.377；No. 3 peaks: 0.394~0.401；No. 4 peaks: 0.498~0.501；No. 5 peaks: 0.520~0.527；No. 6
Peak: 0.544~0.546；No. 7 peaks: 0.626~0.629；No. 8 peaks: 0.701~0.710；No. 9 peaks: 0.708~0.730；No. 10
Peak: 0.867~0.868；No. 11 peaks: 1.000；No. 12 peaks: 1.017~1.021；No. 13 peaks: 1.033~1.050；No. 14 peaks:
1.048~1.075；No. 15 peaks: 1.071~1.087；No. 16 peaks: 1.085~1.099；No. 17 peaks: 1.126~1.129；No. 18
Peak: 1.162~1.167；No. 19 peaks: 1.233~1.238；No. 20 peaks: 1.596~1.606；No. 21 peaks: 1.711~1.723.
As the preferred embodiment of construction method of the present invention, the wide dragon health product characteristic fingerprint pattern includes 20
A common characteristic peaks are referring to peak, the relative retention time of each characteristic peak are as follows: No. 1 peak: 0.365~0.367 with No. 14 peaks；No. 2
Peak: 0.405~0.407；No. 3 peaks: 0.423~0.425；No. 4 peaks: 0.454~0.458；No. 5 peaks: 0.511~0.515；No. 6
Peak: 0.557~0.580；No. 7 peaks: 0.582~0.586；No. 8 peaks: 0.595~0.599；No. 9 peaks: 0.682~0.684；No. 10
Peak: 0.754~0.756；No. 11 peaks: 0.771~0.773；No. 12 peaks: 0.892~0.904；No. 13 peaks: 0.966~0.973；
No. 14 peaks: 1.000；No. 15 peaks: 1.042~1.043；No. 16 peaks: 1.066~1.067；No. 17 peaks: 1.118~1.123；No. 18
Peak: 1.170~1.174；No. 19 peaks: 1.263~1.268；No. 20 peaks: 1.717~1.723.
Second aspect, the present invention provides the wide dragon of above-mentioned construction method building or the characteristic fingerprint patterns of its processed product
Application in wide dragon health product or the detection of its processed product.
The third aspect, the present invention provides a kind of wide dragon or the detection methods of its processed product, including by wide dragon health product
Or the characteristic spectrum of its processed product and construction method as described in any one of claims 1 to 7 building wide dragon health product or its
The characteristic fingerprint pattern of processed product is compared.
Preferred embodiment as detection method of the present invention, comprising the following steps:
(1) wide dragon health product or its processed product are crushed, NaCl solution, ultrasonic extraction is added, centrifugation takes supernatant, uses
NaCl solution constant volume, filtering, obtains test solution；Using high performance liquid chromatography detect to obtain wide dragon health product to be measured or its
Processed product characteristic spectrum, the chromatographic condition are as follows:
Chromatographic column: C18 column；
Mobile phase: using 0.01mol potassium dihydrogen phosphate as mobile phase A, ladder is carried out by Mobile phase B of 70% methanol solution
Degree elution, the eluent gradient elution requirement are as follows: 0~5min, A in mobile phase (100%, B (0%) in mobile phase；5~
20min, A (100% → 85%~80%) in mobile phase, B (0% → 15%~20%) in mobile phase；20~30min, flowing
A (85%~80% → 65%~50%) in phase, B (15%~20% → 35%~50%) in mobile phase；30~40min, stream
Move A (65%~50% → 0%) in phase, B (35%~50% → 100%) in mobile phase；40~45min, A in mobile phase
(0%), B (100%) in mobile phase；
Detector: UV detector；
Flow velocity: 0.8~1.0mL/min；
Detection wavelength: 250~260nm；
Column temperature: 20~25 DEG C；
It is accurate respectively to draw each 10~15 μ L of test solution and control solution, Ultra Performance Liquid Chromatography instrument is injected, is surveyed
It is fixed；
(2) by the resulting wide dragon health product to be measured of step (1) or its processed product characteristic spectrum and wide dragon health product or its big gun
The characteristic fingerprint pattern of product is compared；
The wide dragon processed product characteristic fingerprint pattern includes 21 common characteristic peaks, wherein and No. 1 peak is uracil, No. 7
Peak is hypoxanthine, and No. 11 peaks are inosine, other are non-principal component peak；It is referring to peak, the phase of each characteristic peak with No. 11 peaks of inosine
To retention time are as follows: No. 1 peak: 0.356~0.362；No. 2 peaks: 0.374~0.377；No. 3 peaks: 0.394~0.401；No. 4 peaks:
0.498~0.501；No. 5 peaks: 0.520~0.527；No. 6 peaks: 0.544~0.546；No. 7 peaks: 0.626~0.629；No. 8 peaks:
0.701~0.710；；No. 9 peaks: 0.708~0.730；No. 10 peaks: 0.867~0.868；No. 11 peaks: 1.000；No. 12 peaks:
1.017~1.021；No. 13 peaks: 1.033~1.050；No. 14 peaks: 1.048~1.075；No. 15 peaks: 1.071~1.087；No. 16
Peak: 1.085~1.099；No. 17 peaks: 1.126~1.129；No. 18 peaks: 1.162~1.167；No. 19 peaks: 1.233~1.238；
No. 20 peaks: 1.596~1.606；No. 21 peaks: 1.711~1.723；
The wide dragon health product characteristic fingerprint pattern includes 20 common characteristic peaks, is referring to peak, each feature with No. 14 peaks
The relative retention time at peak are as follows: No. 1 peak: 0.365~0.367；No. 2 peaks: 0.405~0.407；No. 3 peaks: 0.423~0.425；4
Number peak: 0.454~0.458；No. 5 peaks: 0.511~0.515；No. 6 peaks: 0.557~0.580；No. 7 peaks: 0.582~0.586；8
Number peak: 0.595~0.599；No. 9 peaks: 0.682~0.684；No. 10 peaks: 0.754~0.756；No. 11 peaks: 0.771~0.773；
No. 12 peaks: 0.892~0.904；No. 13 peaks: 0.966~0.973；No. 14 peaks: 1.000；No. 15 peaks: 1.042~1.043；No. 16
Peak: 1.066~1.067；No. 17 peaks: 1.118~1.123；No. 18 peaks: 1.170~1.174；No. 19 peaks: 1.263~1.268；
No. 20 peaks: 1.717~1.723；
(3) wide dragon health product to be measured or the quality and/or the true and false of its processed product are determined according to the comparison result of step (2).
As the preferred embodiment of detection method of the present invention, criterion in the step (3) are as follows: such as to be measured wide
The test solution characteristic spectrum of pheretima health product or its processed product meets wide dragon health product or the characteristic fingerprint pattern of its processed product,
It then can determine that wide dragon health product to be measured or its processed product be qualified samples；Conversely, then can determine that wide dragon health product to be measured or its big gun
Product is failed test sample.
Compared with prior art, the invention has the benefit that
(1) the high performance liquid chromatography characteristic fingerprint pattern of wide dragon health product of the invention has 20 shared peaks, shares peak
It is 80% or more that area, which accounts for total peak area,；The high performance liquid chromatography characteristic fingerprint pattern of wide dragon processed product has 21 shared peaks,
It is 80% or more that the area at shared peak, which accounts for total peak area,；The present invention supplements existing wide dragon finger-print, enables it
Quality testing is evidence-based, and the principle active component in wide dragon can be gone out with complete detection, mentions for wide dragon quality overall evaluation
Standard for reference more effectively carries out quality monitoring to medicinal material；
(2) present invention uses hypoxanthine, inosine, uracil as reference substance solution for main Testing index, can be effective
The miscellaneous peak in elution process is detected, wide dragon health product and processed product can accurately be calculated；
(3) the method for the present invention is easy to operate, stable, reproducible, can carry out more to wide dragon health product and processed product
Perfect quality evaluation can rapidly and accurately identify wide dragon health product and processed product, and this method result is accurate, and precision is high,
And save reagent；
Detailed description of the invention
Fig. 1 is the high-efficiency liquid-phase fingerprint and its reference fingerprint of 11 batches of wide dragon health products.
Fig. 2 is the high-efficiency liquid-phase fingerprint and its reference fingerprint of 11 batches of wide dragon processed products.
Fig. 3 is the finger-print of 11 batches of wide dragon processed product reference fingerprints and reference substance solution.
Specific embodiment
Purposes, technical schemes and advantages in order to better illustrate the present invention, below in conjunction with the drawings and specific embodiments pair
The present invention is described further.
The construction method of 1 wide dragon health product of embodiment or its processed product finger-print
1. test material and instrument
(1) material
Hypoxanthine (20161219-2, Baoji time Biotechnology Co., Ltd), inosine (140669-201606, in
State's food and medicine examine and determine research institute), uracil (100469-201302, National Institute for Food and Drugs Control).11 batches of wide dragons
Health product source situation is as shown in table 1, and the processed product source situation that 11 batches of Radix Glycyrrhizaes process wide dragon is as shown in table 2, and through brilliant pair
Professor (Traditional Chinese Medicine University Of Guangzhou) is accredited as Ju Yin section animal Pheretima aspergillum Pheretima aspergillum (E.Perrier)
Hirudo leech.
Table 1
Table 2
| Number | Processed product lot number | Date of manufacture |
| 1 | YPA7J0001 | 2017.10 |
| 2 | YPA7J0002 | 2017.10 |
| 3 | YPA7J0003 | 2017.10 |
| 4 | YPA7J0004 | 2017.10 |
| 5 | YPA7J0005 | 2017.10 |
| 6 | YPA7J0006 | 2017.10 |
| 7 | YPA7J0007 | 2017.10 |
| 8 | YPA7J0008 | 2017.10 |
| 9 | YPA7J0009 | 2017.10 |
| 10 | YPA7J0010 | 2017.10 |
| 11 | YPA7J0011 | 2017.10 |
(2) reagent and instrument
Methanol, acetonitrile are chromatographically pure, and water is ultrapure water, and other reagents are that analysis is pure.
High performance liquid chromatograph (UltiMate3000 type, Dai An company of the U.S.), ultralow organic type ultrapure water machine (Guangzhou
Hua Chun Water Management Equipment Ltd.), CQ-150A is cleaned by ultrasonic in instrument (Shanghai leap medical optical equipment factory), low speed it is desk-top from
Scheming (Beijing epoch Bei Li centrifuge Co., Ltd).
(3) data processing software
Chromatographic fingerprints of Chinese materia medica similarity evaluation system software (2004A editions, Chinese Pharmacopoeia Commission).
2. test method
(1) prepared by test solution
Each sample powder about 0.5g is taken, it is accurately weighed, it is placed in 100mL tool plug triangular pyramidal bottle, is separately added into 0.9%
NaCl solution 10mL, ultrasonic extraction 30min, 5000rmin-1It is centrifuged 10min, supernatant is taken, with 0.9%NaCl solution constant volume
To 10mL, 0.45 μm of miillpore filter is filtered to obtain the final product.
(2) prepared by reference substance solution
Take hypoxanthine, inosine, uracil reference substance appropriate respectively, it is accurately weighed, it sets in 10mL measuring bottle, with 0.9%
NaCl solution is made into the reference substance of every milliliter of 0.0897mg containing hypoxanthine, inosine 0.0960mg, uracil 0.0992mg respectively
Solution.Reference substance solution saved at 4 DEG C to get.
(3) chromatographic condition
Chromatographic column: Innoval ODS-2C18 column (5 μm of 250mm × 4.6mm)；
Detection wavelength: 254nm；
20 DEG C of column temperature；
Flow velocity: 0.8mL/min；
10 μ L of sample volume,
Mobile phase: using 0.01mol potassium dihydrogen phosphate as mobile phase A, ladder is carried out by Mobile phase B of 70% methanol solution
Degree elution, eluent gradient elution requirement are as follows: 0~5min, A (100%) in mobile phase, B (0%) in mobile phase；5~20min,
A (100% → 80%) in mobile phase, B (0% → 20%) in mobile phase；20~30min, A (80% → 50%) in mobile phase,
B (20% → 50%) in mobile phase；30~40min, A (50% → 0%) in mobile phase, B (50% → 100%) in mobile phase；
40~45min, A (0%) in mobile phase, B (100%) in mobile phase；
(4) methodological study
Precision: YPA7J0001 batch wide dragon processed product is prepared into test solution, is connected with high performance liquid chromatograph
It continuous sample introduction five times, is analyzed after handling data by similarity software, the method for selecting Supplements and median,
The similarity result obtained is less than in the relative retention time RSD that 0.99 or more, five sample introduction finger-print shares peak
0.1%, the RSD value of relative peak area prompts instrument stabilizer, there is good precision less than 3.0%.
Stability: being prepared into test solution for YPA7J0001 batch wide dragon processed product, with high performance liquid chromatograph point
Not 0 hour, 6 hours, 12 hours, 18 hours, 24 hours this five time points difference sample introductions it is primary, each main color as the result is shown
For the RSD of the relative retention time of spectral peak less than 1%, the RSD of relative peak area prompts test solution at 24 hours less than 3%
It is interior with good stability.
Repeatability: YPA7J0001 batch wide dragon processed product is prepared into 6 parts of test solutions, uses high performance liquid chromatography
Instrument difference sample introduction is primary, as a result prompts the RSD of the relative retention time of each main chromatographic peak less than 0.2%, relative peak area
RSD prompts this method to have good repeatability less than 3.0%.
(5) sample determining fingerprint pattern
The test solution for taking each batch wide dragon health product and wide dragon processed product to be prepared into respectively, is placed in efficient liquid phase instrument
It in autosampler, is measured by the chromatographic condition of the present embodiment, passes through chromatographic fingerprints of Chinese materia medica similarity evaluation system
Software (2004A editions, Chinese Pharmacopoeia Commission) processing are to get 11 batch health product wide dragon high-efficiency liquid-phase fingerprints and its control
Finger-print is as shown in Figure 1, wherein and S1~S11 is 1~11 batch of health product wide dragon finger-print, and S12 is reference fingerprint,
Supplements are carried out by similarity evaluation, Auto-matching generates；11 batch wide dragon processed products
High-efficiency liquid-phase fingerprint and its reference fingerprint are as shown in Figure 2, wherein S1~S11 is that 1~11 batch of wide dragon processed product refers to
Line map, S12 are reference fingerprint, carry out Supplements, automatic by similarity evaluation
With generation；11 batches of wide dragon processed product reference fingerprints and the finger-print of reference substance solution are as shown in Figure 3, wherein S1 is
Hypoxanthine reference substance finger-print, S2 are inosine reference substance finger-print, and S3 is uracil reference substance finger-print, S4 11
Criticize wide dragon processed product reference fingerprint.
3. test result
(1) fingerprint map analyzing
Retention time is measured to 11 batches of wide dragon health products and wide dragon processed product using high performance liquid chromatography, with inosine chromatography
Peak is to calculate relative retention time referring to peak.Then shared peak is demarcated using relative retention time, to 11 batches of wide dragons
The finger-print of health product is matched, and demarcates 20 shared peaks altogether, and shared peak area accounts for the 80.8% of total peak area, as a result such as table
Shown in 3；The finger-print of 11 batches of wide dragon processed products is matched, demarcates 21 shared peaks altogether, shared peak area accounts for Zong Feng
The 81.3% of area, the results are shown in Table 4.
Table 3
Table 4
Characteristic fingerprint pattern determines: by chromatographic peak appearance time in reference substance solution and test solution chromatogram and
Chromatographic peak obtains the common characteristic peaks in test solution map with the presence or absence of being compared；To 11 batches of wide dragon processed product samples
Product are detected, the wide dragon being made of using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 21 common characteristic peaks
Processed product characteristic fingerprint pattern；11 batches of wide dragon health product samples are detected, are commented using chromatographic fingerprints of Chinese materia medica similarity
The wide dragon health product characteristic fingerprint pattern that valence Software Create is made of 20 common characteristic peaks.
Characteristic fingerprint pattern description:
Wide dragon processed product characteristic fingerprint pattern includes 21 common characteristic peaks, wherein No. 1 peak is uracil, and No. 7 peaks are
Hypoxanthine, No. 11 peaks are inosine, other are non-principal component peak；It is referring to peak, the opposite guarantor of each characteristic peak with No. 11 peaks of inosine
Stay the time are as follows: No. 1 peak: 0.356~0.362；No. 2 peaks: 0.374~0.377；No. 3 peaks: 0.394~0.401；No. 4 peaks: 0.498
~0.501；No. 5 peaks: 0.520~0.527；No. 6 peaks: 0.544~0.546；No. 7 peaks: 0.626~0.629；No. 8 peaks: 0.701
~0.710；No. 9 peaks: 0.708~0.730；No. 10 peaks: 0.867~0.868；No. 11 peaks: 1.000；No. 12 peaks: 1.017~
1.021；No. 13 peaks: 1.033~1.050；No. 14 peaks: 1.048~1.075；No. 15 peaks: 1.071~1.087；No. 16 peaks:
1.085~1.099；No. 17 peaks: 1.126~1.129；No. 18 peaks: 1.162~1.167；No. 19 peaks: 1.233~1.238；No. 20
Peak: 1.596~1.606；No. 21 peaks: 1.711~1.723.
Wide dragon health product characteristic fingerprint pattern includes 20 common characteristic peaks, is reference peak with No. 14 peaks, each characteristic peak
Relative retention time are as follows: No. 1 peak: 0.365~0.367；No. 2 peaks: 0.405~0.407；No. 3 peaks: 0.423~0.425；No. 4
Peak: 0.454~0.458；No. 5 peaks: 0.511~0.515；No. 6 peaks: 0.557~0.580；No. 7 peaks: 0.582~0.586；No. 8
Peak: 0.595~0.599；No. 9 peaks: 0.682~0.684；No. 10 peaks: 0.754~0.756；No. 11 peaks: 0.771~0.773；12
Number peak: 0.892~0.904；No. 13 peaks: 0.966~0.973；No. 14 peaks: 1.000；No. 15 peaks: 1.042~1.043；No. 16
Peak: 1.066~1.067；No. 17 peaks: 1.118~1.123；No. 18 peaks: 1.170~1.174；No. 19 peaks: 1.263~1.268；
No. 20 peaks: 1.717~1.723.
(2) similarity evaluation
The fingerprint similarity evaluation system software promulgated using generally acknowledged Chinese Pharmacopoeia Commission is to obtained
Wide dragon processed product and its health product finger-print are handled, as a result respectively as shown in table 5, table 6.
Table 5
Table 6
By table 5 and table 6 it is found that 11 batches of wide dragon processed product similarities are between 0.871~0.980,11 batches of wide dragons are raw
For product similarity between 0.895~0.991, health product and processed product similarity are higher, illustrate this 11 batches of wide dragon health products and processing
Product sample room difference is smaller, and quality is stablized.
Embodiment 2
A kind of embodiment of the construction method of the characteristic fingerprint pattern of wide dragon of the present invention or its processed product, including following step
It is rapid:
(1) preparation of test solution: crushed 80 meshes for wide dragon processed product or its health product, accurately weighed, set tool
It fills in conical flask, is separately added into 0.9%NaCl solution 10mL, ultrasonic extraction 20min, 5000rmin-1It is centrifuged 10min, is taken
Clear liquid is settled to 10mL with 0.9%NaCl solution, filters to obtain the final product, in test solution wide dragon health product or its processed product with
The mass volume ratio of NaCl solution is 1:20.
(2) preparation of reference substance solution: precision weighing hypoxanthine, inosine, uracil reference substance add 0.9%NaCl molten
Liquid dissolution, as reference substance solution, hypoxanthic concentration is 80 μ g/ml in the reference substance solution, the concentration of inosine is 80 μ
G/ml, uracil concentration be 80 μ g/ml.
(3) accurate respectively to draw test solution and control solution, it injects in Ultra Performance Liquid Chromatography instrument and is measured,
Obtain the wide dragon processed product finger-print being made of 21 peaks；The wide dragon health product finger-print being made of 20 peaks；
Chromatographic condition are as follows:
Chromatographic column: C18 column；Detector: UV detector；Flow velocity: 1.0mL/min；Detection wavelength: 260nm；Column temperature 25
℃；15 μ L of sample volume；Mobile phase A is 0.01mol potassium dihydrogen phosphate, and Mobile phase B is 70% methanol solution；Gradient elution journey
Sequence are as follows:
0~5min, mobile phase A: 100%, Mobile phase B: 0%；
5~20min, mobile phase A: 100% → 80%, Mobile phase B: 0% → 20%；
20~30min, mobile phase A: 80% → 65%, Mobile phase B: 20% → 35%；
30~40min, mobile phase A: 65% → 0%, Mobile phase B: 35% → 100%；
40~45min, mobile phase A: 0%, Mobile phase B: 100%.
The method for constructing wide dragon or the characteristic fingerprint pattern of its processed product are as follows: pass through reference substance solution and test solution
Chromatographic peak appearance time and chromatographic peak in chromatogram obtain the shared spy in test solution map with the presence or absence of being compared
Levy peak；11 batches of wide dragon processed product samples are detected, using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create by
The wide dragon processed product characteristic fingerprint pattern that 21 common characteristic peaks are constituted；11 batches of wide dragon health product samples are detected, are adopted
The wide dragon health product characteristic fingerprint being made of with chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 20 common characteristic peaks
Map.
The wide dragon health product of the present embodiment building or the characteristic fingerprint pattern of processed product are the same as embodiment 1.
Embodiment 3
A kind of embodiment of the construction method of the characteristic fingerprint pattern of wide dragon of the present invention or its processed product, including following step
It is rapid:
(1) prepared by test solution: wide dragon processed product or its health product be crushed into 80 meshes, it is accurately weighed, and set tool plug
In conical flask, it is separately added into 0.9%NaCl solution 10mL, ultrasonic extraction 20min, 5000rmin-1It is centrifuged 10min, takes supernatant
Liquid is settled to 10mL with 0.9%NaCl solution, filters to obtain the final product, wide dragon health product or its processed product and NaCl in test solution
The mass volume ratio of solution is 1:20.
(2) prepared by reference substance solution: precision weighing hypoxanthine, inosine, uracil reference substance add 0.9%NaCl solution
Dissolution, as reference substance solution, hypoxanthic concentration is 95 μ g/ml in the reference substance solution, the concentration of inosine is 98 μ g/
Ml, uracil concentration be 117 μ g/ml.
(3) accurate respectively to draw test solution and control solution, it injects in Ultra Performance Liquid Chromatography instrument and is measured,
Obtain the wide dragon processed product finger-print being made of 21 peaks；The wide dragon health product finger-print being made of 20 peaks；
Chromatographic condition are as follows:
Chromatographic column: C18 column；Detector: UV detector；Flow velocity: 1.0mL/min；Detection wavelength: 250nm；Column temperature 20
℃；10 μ L of sample volume；Mobile phase A is 0.01mol potassium dihydrogen phosphate, and Mobile phase B is 70% methanol solution；Gradient elution journey
Sequence are as follows:
0~5min, mobile phase A: 100%, Mobile phase B: 0%；
5~20min, mobile phase A: 100% → 85%, Mobile phase B: 0% → 15%；
20~30min, mobile phase A: 85% → 50%, Mobile phase B: 15% → 50%；
30~40min, mobile phase A: 50% → 0%, Mobile phase B: 50% → 100%；
40~45min, mobile phase A: 0%, Mobile phase B: 100%.
The method for constructing wide dragon or the characteristic fingerprint pattern of its processed product are as follows: pass through reference substance solution and test solution
Chromatographic peak appearance time and chromatographic peak in chromatogram obtain the shared spy in test solution map with the presence or absence of being compared
Levy peak；11 batches of wide dragon processed product samples are detected, using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create by
The wide dragon processed product characteristic fingerprint pattern that 21 common characteristic peaks are constituted；11 batches of wide dragon health product samples are detected, are adopted
The wide dragon health product characteristic fingerprint being made of with chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 20 common characteristic peaks
Map.
The wide dragon health product of the present embodiment building or the characteristic fingerprint pattern of processed product are the same as embodiment 1.
Embodiment 4
A kind of embodiment of the construction method of the characteristic fingerprint pattern of wide dragon of the present invention or its processed product, including following step
It is rapid:
(1) preparation of test solution: wide dragon processed product or its health product are crushed, accurately weighed, set stuffed conical flask
In, it is separately added into 0.9%NaCl solution 15mL, ultrasonic extraction 30min, 5000rmin-1It is centrifuged 20min, takes supernatant, is used
0.9%NaCl solution is settled to 20mL, filters to obtain the final product, wide dragon health product or its processed product and NaCl solution in test solution
Mass volume ratio is 1:30.
(2) preparation of reference substance solution: precision weighing hypoxanthine, inosine, uracil reference substance add 0.9%NaCl molten
Liquid dissolution, as reference substance solution, hypoxanthic concentration is 100 μ g/ml in the reference substance solution, the concentration of inosine is
100 μ g/ml, uracil concentration be 120 μ g/ml.
(3) accurate respectively to draw test solution and control solution, it injects in Ultra Performance Liquid Chromatography instrument and is measured,
Obtain the wide dragon processed product finger-print being made of 21 peaks；The wide dragon health product finger-print being made of 20 peaks；
Chromatographic condition are as follows:
Chromatographic column: C18 column；Detector: UV detector；Flow velocity: 0.8mL/min；Detection wavelength: 255nm；Column temperature 22
℃；15 μ L of sample volume；Mobile phase A is 0.01mol potassium dihydrogen phosphate, and Mobile phase B is 70% methanol solution；Gradient elution journey
Sequence are as follows:
0~5min, mobile phase A: 100%, Mobile phase B: 0%；
5~20min, mobile phase A: 100% → 85%, Mobile phase B: 0% → 15%；
20~30min, mobile phase A: 85% → 55%, Mobile phase B: 15% → 45%；
30~40min, mobile phase A: 55% → 0%, Mobile phase B: 45% → 100%；
40~45min, mobile phase A: 0%, Mobile phase B: 100%.
The method for constructing wide dragon or the characteristic fingerprint pattern of its processed product are as follows: pass through reference substance solution and test solution
Chromatographic peak appearance time and chromatographic peak in chromatogram obtain the shared spy in test solution map with the presence or absence of being compared
Levy peak；11 batches of wide dragon processed product samples are detected, using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create by
The wide dragon processed product characteristic fingerprint pattern that 21 common characteristic peaks are constituted；11 batches of wide dragon health product samples are detected, are adopted
The wide dragon health product characteristic fingerprint being made of with chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 20 common characteristic peaks
Map.
The wide dragon health product of the present embodiment building or the characteristic fingerprint pattern of processed product are the same as embodiment 1.
Embodiment 5
A kind of embodiment of wide dragon of the present invention or its processed product detection method, comprising the following steps:
(1) using high-efficiency liquid chromatography method for detecting same as Example 2 detect to obtain wide dragon health product to be measured or its
Processed product characteristic spectrum；
(2) by the resulting wide dragon health product to be measured of step (1) or its processed product characteristic spectrum and wide dragon health product or its big gun
The characteristic fingerprint pattern of product is compared；
(3) wide dragon health product to be measured or the quality and/or the true and false of its processed product are determined according to the comparison result of step (2).
Embodiment 6
A kind of embodiment of wide dragon of the present invention or its processed product detection method, comprising the following steps:
(1) using high-efficiency liquid chromatography method for detecting same as Example 3 detect to obtain wide dragon health product to be measured or its
Processed product characteristic spectrum；
(2) by the resulting wide dragon health product to be measured of step (1) or its processed product characteristic spectrum and wide dragon health product or its big gun
The characteristic fingerprint pattern of product is compared；
(3) wide dragon health product to be measured or the quality and/or the true and false of its processed product are determined according to the comparison result of step (2).
Embodiment 7
A kind of embodiment of wide dragon of the present invention or its processed product detection method, comprising the following steps:
(1) using high-efficiency liquid chromatography method for detecting same as Example 4 detect to obtain wide dragon health product to be measured or its
Processed product characteristic spectrum；
(2) by the resulting wide dragon health product to be measured of step (1) or its processed product characteristic spectrum and wide dragon health product or its big gun
The characteristic fingerprint pattern of product is compared；
(3) wide dragon health product to be measured or the quality and/or the true and false of its processed product are determined according to the comparison result of step (2).
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range is protected, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should
Understand, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the essence of technical solution of the present invention
And range.
Claims (10)
1. a kind of construction method of the characteristic fingerprint pattern of wide dragon or its processed product, which comprises the following steps:
(1) prepared by test solution: wide dragon health product or its processed product are crushed, NaCl solution is added, ultrasonic extraction is centrifuged,
Supernatant is taken, with NaCl solution constant volume, filtering obtains test solution；
(2) prepared by reference substance solution: taking hypoxanthine, inosine, uracil reference substance, adds NaCl solution to dissolve, obtain reference substance
Solution；
(3) high performance liquid chromatography detection constructs wide dragon health product or the characteristic fingerprint pattern of its processed product.
2. construction method as described in claim 1, which is characterized in that in the step (1), the concentration of NaCl solution is
0.9%, the time of ultrasonic extraction is 20~30min, centrifugal rotational speed 5000rmin-1, centrifugation time is 10~20min, is supplied
Wide dragon health product or its processed product and the mass volume ratio of NaCl solution are 1:20 in test sample solution.
3. construction method as described in claim 1, which is characterized in that in the step (2), the concentration of NaCl solution is
0.9%, hypoxanthic concentration is 80~100 μ g/mL in reference substance solution, and the concentration of inosine is 80~100 μ g/mL, is urinated phonetic
The concentration of pyridine is 80~120 μ g/mL.
4. construction method as described in claim 1, which is characterized in that chromatographic condition in the step (3) are as follows:
Chromatographic column: C18 column；
Mobile phase: using 0.01mol potassium dihydrogen phosphate as mobile phase A, gradient is carried out as Mobile phase B using 70% methanol solution and is washed
It is de-, the eluent gradient elution requirement are as follows: 0~5min, A (100%) in mobile phase, B (0%) in mobile phase；5~20min,
A (100% → 85%~80%) in mobile phase, B (0% → 15%~20%) in mobile phase；20~30min, A in mobile phase
(85%~80% → 65%~50%), B (15%~20% → 35%~50%) in mobile phase；30~40min, in mobile phase
A (65%~50% → 0%), B (35%~50% → 100%) in mobile phase；40~45min, A (0%) in mobile phase, flowing
B (100%) in phase；
Detector: UV detector；
Flow velocity: 0.8~1.0mL/min；
Detection wavelength: 250~260nm；
Column temperature: 20~25 DEG C；
It is accurate respectively to draw each 10~15 μ L of test solution and control solution, inject high performance liquid chromatograph, measurement.
5. construction method as described in claim 1, which is characterized in that building wide dragon or its processed product in the step (3)
Characteristic fingerprint pattern method are as follows: pass through the chromatographic peak appearance time and color in reference substance solution and test solution chromatogram
Spectral peak obtains the common characteristic peaks in test solution map with the presence or absence of being compared；To 11 batches of wide dragon processed product samples
It is detected, the wide dragon big gun being made of using chromatographic fingerprints of Chinese materia medica similarity evaluation Software Create 21 common characteristic peaks
Product characteristics finger-print；11 batches of wide dragon health product samples are detected, using chromatographic fingerprints of Chinese materia medica similarity evaluation
The wide dragon health product characteristic fingerprint pattern that Software Create is made of 20 common characteristic peaks.
6. construction method as claimed in claim 5, which is characterized in that the wide dragon processed product characteristic fingerprint pattern includes 21
A common characteristic peaks, wherein No. 1 peak is uracil, and No. 7 peaks are hypoxanthine, and No. 11 peaks are inosine, other are non-principal component
Peak；
It is referring to peak, the relative retention time of each characteristic peak are as follows: No. 1 peak: 0.356~0.362 with No. 11 peaks of inosine；No. 2 peaks:
0.374~0.377；No. 3 peaks: 0.394~0.401；No. 4 peaks: 0.498~0.501；No. 5 peaks: 0.520~0.527；No. 6 peaks:
0.544~0.546；No. 7 peaks: 0.626~0.629；No. 8 peaks: 0.701~0.710；No. 9 peaks: 0.708~0.730；No. 10 peaks:
0.867~0.868；No. 11 peaks: 1.000；No. 12 peaks: 1.017~1.021；No. 13 peaks: 1.033~1.050；No. 14 peaks:
1.048~1.075；No. 15 peaks: 1.071~1.087；No. 16 peaks: 1.085~1.099；No. 17 peaks: 1.126~1.129；No. 18
Peak: 1.162~1.167；No. 19 peaks: 1.233~1.238；No. 20 peaks: 1.596~1.606；No. 21 peaks: 1.711~1.723.
7. construction method as claimed in claim 5, which is characterized in that the wide dragon health product characteristic fingerprint pattern includes 20
Common characteristic peaks are referring to peak, the relative retention time of each characteristic peak are as follows: No. 1 peak: 0.365~0.367 with No. 14 peaks；No. 2
Peak: 0.405~0.407；No. 3 peaks: 0.423~0.425；No. 4 peaks: 0.454~0.458；No. 5 peaks: 0.511~0.515；No. 6
Peak: 0.557~0.580；No. 7 peaks: 0.582~0.586；No. 8 peaks: 0.595~0.599；No. 9 peaks: 0.682~0.684；No. 10
Peak: 0.754~0.756；No. 11 peaks: 0.771~0.773；No. 12 peaks: 0.892~0.904；No. 13 peaks: 0.966~0.973；
No. 14 peaks: 1.000；No. 15 peaks: 1.042~1.043；No. 16 peaks: 1.066~1.067；No. 17 peaks: 1.118~1.123；No. 18
Peak: 1.170~1.174；No. 19 peaks: 1.263~1.268；No. 20 peaks: 1.717~1.723.
8. the wide dragon of construction method building as described in any one of claims 1 to 7 or the characteristic fingerprint pattern of its processed product
Application in wide dragon health product or the detection of its processed product.
9. the detection method of a kind of wide dragon or its processed product, which is characterized in that including by wide dragon health product or its processed product
The feature of the wide dragon health product or its processed product of characteristic spectrum and construction method as described in any one of claims 1 to 7 building
Finger-print is compared.
10. detection method as claimed in claim 9, which comprises the following steps:
(1) wide dragon health product or its processed product are crushed, NaCl solution, ultrasonic extraction is added, centrifugation takes supernatant, uses NaCl
Solution constant volume, filtering, obtains test solution；It detects to obtain wide dragon health product to be measured or its processing using high performance liquid chromatography
Product characteristic spectrum, the chromatographic condition are as follows:
Chromatographic column: C18 column；
Mobile phase: using 0.01mol potassium dihydrogen phosphate as mobile phase A, gradient is carried out as Mobile phase B using 70% methanol solution and is washed
It is de-, the eluent gradient elution requirement are as follows: 0~5min, A in mobile phase (100%, B (0%) in mobile phase；5~20min,
A (100% → 85%~80%) in mobile phase, B (0% → 15%~20%) in mobile phase；20~30min, A in mobile phase
(85%~80% → 65%~50%), B (15%~20% → 35%~50%) in mobile phase；30~40min, in mobile phase
A (65%~50% → 0%), B (35%~50% → 100%) in mobile phase；40~45min, A (0%) in mobile phase, flowing
B (100%) in phase；
Detector: UV detector；
Flow velocity: 0.8~1.0mL/min；
Detection wavelength: 250~260nm；
Column temperature: 20~25 DEG C；
It is accurate respectively to draw each 10~15 μ L of test solution and control solution, inject Ultra Performance Liquid Chromatography instrument, measurement；
(2) by the resulting wide dragon health product to be measured of step (1) or its processed product characteristic spectrum and wide dragon health product or its processed product
Characteristic fingerprint pattern be compared；
The wide dragon processed product characteristic fingerprint pattern includes 21 common characteristic peaks, wherein No. 1 peak is uracil, and No. 7 peaks are
Hypoxanthine, No. 11 peaks are inosine, other are non-principal component peak；It is referring to peak, the opposite guarantor of each characteristic peak with No. 11 peaks of inosine
Stay the time are as follows: No. 1 peak: 0.356~0.362；No. 2 peaks: 0.374~0.377；No. 3 peaks: 0.394~0.401；No. 4 peaks: 0.498
~0.501；No. 5 peaks: 0.520~0.527；No. 6 peaks: 0.544~0.546；No. 7 peaks: 0.626~0.629；No. 8 peaks: 0.701
~0.710；；No. 9 peaks: 0.708~0.730；No. 10 peaks: 0.867~0.868；No. 11 peaks: 1.000；No. 12 peaks: 1.017~
1.021；No. 13 peaks: 1.033~1.050；No. 14 peaks: 1.048~1.075；No. 15 peaks: 1.071~1.087；No. 16 peaks:
1.085~1.099；No. 17 peaks: 1.126~1.129；No. 18 peaks: 1.162~1.167；No. 19 peaks: 1.233~1.238；No. 20
Peak: 1.596~1.606；No. 21 peaks: 1.711~1.723；
The wide dragon health product characteristic fingerprint pattern includes 20 common characteristic peaks, is reference peak with No. 14 peaks, each characteristic peak
Relative retention time are as follows: No. 1 peak: 0.365~0.367；No. 2 peaks: 0.405~0.407；No. 3 peaks: 0.423~0.425；No. 4
Peak: 0.454~0.458；No. 5 peaks: 0.511~0.515；No. 6 peaks: 0.557~0.580；No. 7 peaks: 0.582~0.586；No. 8
Peak: 0.595~0.599；No. 9 peaks: 0.682~0.684；No. 10 peaks: 0.754~0.756；No. 11 peaks: 0.771~0.773；12
Number peak: 0.892~0.904；No. 13 peaks: 0.966~0.973；No. 14 peaks: 1.000；No. 15 peaks: 1.042~1.043；No. 16
Peak: 1.066~1.067；No. 17 peaks: 1.118~1.123；No. 18 peaks: 1.170~1.174；No. 19 peaks: 1.263~1.268；
No. 20 peaks: 1.717~1.723；
(3) wide dragon health product to be measured or the quality and/or the true and false of its processed product are determined according to the comparison result of step (2).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114047268A (en) * | 2021-11-26 | 2022-02-15 | 华润三九医药股份有限公司 | Fingerprint spectrum of Guangdong earthworm medicine preparation, construction method and content determination method thereof |
| CN115078588A (en) * | 2022-07-14 | 2022-09-20 | 江阴天江药业有限公司 | Aspongopus and quality evaluation method of processed product thereof |
| CN115236212A (en) * | 2021-04-22 | 2022-10-25 | 中国科学院上海药物研究所 | Quality detection method of euphorbia Chinese herbal medicine |
| CN115980213A (en) * | 2022-12-20 | 2023-04-18 | 天地恒一制药股份有限公司 | Construction method and application of HPLC (high Performance liquid chromatography) characteristic spectrum of ootheca Mantidis medicinal material, decoction pieces, standard decoction, dry extract powder or formula granules |
| CN115980213B (en) * | 2022-12-20 | 2024-07-02 | 天地恒一制药股份有限公司 | Construction method and application of HPLC characteristic spectrum of ootheca Mantidis medicinal materials, decoction pieces, standard decoction, dry extract powder or formula granule |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844912A (en) * | 2006-01-24 | 2006-10-11 | 李振国 | Earthworm fingerprint spectrum establishment method and medicinal earthworm identification method |
| CN1982892A (en) * | 2006-01-24 | 2007-06-20 | 李振国 | Method and use for setting up Shuxuetong injection or its intermediate fingerprint atlas |
| CN103926350A (en) * | 2014-05-04 | 2014-07-16 | 昆明赛诺制药有限公司 | Inspection method of rehabilitation liquid formulation fingerprint and standard fingerprint |
| CN106370739A (en) * | 2016-08-18 | 2017-02-01 | 四川好医生攀西药业有限责任公司 | Kangfuxin solution preparation fingerprint quality determination method and standard fingerprint |
-
2018
- 2018-12-18 CN CN201811549182.2A patent/CN109324141B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844912A (en) * | 2006-01-24 | 2006-10-11 | 李振国 | Earthworm fingerprint spectrum establishment method and medicinal earthworm identification method |
| CN1982892A (en) * | 2006-01-24 | 2007-06-20 | 李振国 | Method and use for setting up Shuxuetong injection or its intermediate fingerprint atlas |
| CN103926350A (en) * | 2014-05-04 | 2014-07-16 | 昆明赛诺制药有限公司 | Inspection method of rehabilitation liquid formulation fingerprint and standard fingerprint |
| CN106370739A (en) * | 2016-08-18 | 2017-02-01 | 四川好医生攀西药业有限责任公司 | Kangfuxin solution preparation fingerprint quality determination method and standard fingerprint |
Non-Patent Citations (4)
| Title |
|---|
| YI TAO ET AL: "Techniques for biological fingerprinting of traditional Chinese medicine", 《TRENDS IN ANALYTICAL CHEMISTRY》 * |
| 吴文如: "地龙种质资源与品质评价研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 国家药典委员会: "《中国药典2010年版》", 31 January 2010 * |
| 肖会敏 等: "地龙胶囊的HPLC特征图谱和10种成分的定量研究", 《西北药学杂志》 * |
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