CN106370739A - Kangfuxin solution preparation fingerprint quality determination method and standard fingerprint - Google Patents
Kangfuxin solution preparation fingerprint quality determination method and standard fingerprint Download PDFInfo
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- CN106370739A CN106370739A CN201610688619.5A CN201610688619A CN106370739A CN 106370739 A CN106370739 A CN 106370739A CN 201610688619 A CN201610688619 A CN 201610688619A CN 106370739 A CN106370739 A CN 106370739A
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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
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Abstract
The invention relates to a drug detection method and especially relates to a Kangfuxin solution preparation fingerprint quality determination method and a standard fingerprint. Through high performance liquid chromatography detection, a Kangfuxin solution fingerprint is obtained. The method comprises a, contrast solution preparation, b, sample solution preparation and c, fingerprint making. The method builds a Kangfuxin solution chemical fingerprint with good chromatographic peak resolution, repeatability and stability and can be used as a Kangfuxin solution preparation quality control method. The method provides scientific basis for scientific and effective evaluation and control of qualities of periplaneta americana medical raw materials and a Kangfuxin solution preparation.
Description
Technical field
The present invention relates to a kind of measuring method of medicine, particularly to the quality survey of Kangfuxin Liquid preparation finger
Determine method and standard finger-print.
Background technology
Kangfuxin Liquid records in Drug Standard of Ministry of Public Health of the Peoples Republic of China Chinese patent medicine the 19th and ws3-b-3674-
2000(z).The single preparations of ephedrine that Kangfuxin Liquid is made for American-cockroach-extract, have antiinflammatory, anti-peptic ulcer, antitumor,
Multiple pharmacologically active such as immunomodulating, antioxidation.Kangfuxin Liquid clinical practice extensively, is taken orally and be can be used for treatment stomachache bleeding, stomach
And the disease such as duodenal ulcer, external can treat incised wound, wound, ulcer, burn, scald etc..
At present, the quality evaluation of Kangfuxin Liquid many with one or more gradient elutions, aminoacid and polysaccharide containing measurement
Based on fixed research, only minority report has carried out the finger printing research of Kangfuxin Liquid preparation, but the peak type of most of chromatographic peak
All poor with separating degree it is difficult to efficiently control the quality of Kangfuxin Liquid preparation.In order to improve the quality control water of Kangfuxin Liquid
Flat, this research adopts efficient liquid-phase chromatography method, establishes the rehabilitation of most of chromatographic peak separating degree, repeatability and good stability
New liquid chemical fingerprint, and carry out similarity evaluation to the finger printing of different batches sample is science, effectively evaluate and
The inherent quality controlling Kangfuxin Liquid preparation provides foundation.
Content of the invention
It is an object of the invention to disclosing a kind of fingerprint pattern quality determination method of Kangfuxin Liquid preparation, the method has
Good separating effect, sensitivity is high, can truly reflect property material contained by Kangfuxin Liquid preparation, can be used as the Kangfuxin Liquid quality of the pharmaceutical preparations
Control method.
For reaching above-mentioned purpose, technical scheme is as follows:
The measuring method of Kangfuxin Liquid preparation finger of the present invention is it is characterised in that the method includes walking as follows
Rapid:
The preparation of (a) need testing solution: take Kangfuxin Liquid preparation, cross 0.22 μm of microporous filter membrane, take subsequent filtrate, standby；
The preparation of (b) reference substance solution: precision weighs trigonelline, uracil, hypoxanthine, inosine, appropriate adenosine, plus
Water is made every 1ml and is contained trigonelline, uracil, hypoxanthine, inosine and adenosine respectively 20 μ g, 30 μ g, 20 μ g, 40 μ g and 40
The mixing contrast solution of μ g, standby；
C () measures: draw each 20 μ l injection high performance liquid chromatographs of test sample and reference substance solution, with 0.08%～
1.2% acetic acid aqueous solution (a)-methanol (b) is mobile phase, using gradient elution, obtains the finger printing of Kangfuxin Liquid preparation.
Preferably mobile phase is 0.1% acetic acid aqueous solution (a)-methanol (b)
High performance liquid chromatograph condition determination is: filler is octadecylsilane chemically bonded silica；0.08%～1.2% second
Aqueous acid (a)-methanol (b)；Using gradient elution: 0～3min, 3%b；3～5min, 3%b～8%b；5～10min, 8%
B～10%b；10～15min, 10%b～35%b；15～30min, 35%b～80%b；30～40min, 80%b～3%b；
40～45min, 3%b；Detection wavelength is 254nm；Column temperature is 35 DEG C；Flow velocity is 0.8ml/min；Number of theoretical plate presses hypoxanthine
Calculating is not less than 5000.
The fingerprint image of the Kangfuxin Liquid preparation that the present invention obtains is characterised by: Kangfuxin Liquid preparation has 12 total fingers
Stricture of vagina peak, wherein 5 identical with corresponding object of reference peak retention time respectively；Peak corresponding with hypoxanthine object of reference peak is s peak,
The relative retention time of 12 total fingerprint peakses is respectively as follows: No. 1 peak 0.432 ± 0.043,2 peak 0.671 ± 0.067,3 peak
0.731 ± 0.073, No. 4 peaks 0.820 ± 0.082,5 peak 0.888 ± 0.089,6 (s) number peak 1.000 ± 0.000,7 peak
1.126 ± 0.113, No. 8 peaks 1.462 ± 0.146,9 peak 1.536 ± 0.154,10 peak 1.884 ± 0.118,11 peak
2.282 ± 0.228, No. 12 peaks 2.675 ± 0.268.
Advantages of the present invention is as follows:
1. Kangfuxin Liquid preparation finger disclosed by the invention, has carried out Efficient Characterization to its property material, there is provided
A kind of method Kangfuxin Liquid quality evaluated by overall finger printing information, it is to avoid only measure wherein one or two kind change
Study point one-sidedness just the Kangfuxin Liquid quality of the pharmaceutical preparations being judged so that the control of the Kangfuxin Liquid quality of the pharmaceutical preparations is more accurately objective.
2. the fingerprint pattern quality determination method separating degree that the present invention provides is good, repeated and good stability, can be quick
Kangfuxin Liquid is identified.
3. the finger printing that the present invention provides has 12 total peaks, by contrasting with standard substance chromatographic peak, has demarcated chromatograph
5 total peaks of in figure, are respectively as follows: trigonelline, hypoxanthine, uracil, inosine and adenosine.
Specific embodiment:
Following experimental examples and embodiment are used for further illustrating but are not limited to the present invention.
Experimental example 1
(1) instrument and reagent
Tsq quantum ultra liquid chromatography-tandem mass instrument；Hypersil gold c18 post (4.6 ×
150mm, 5 μm).
Methanol is chromatographically pure, and water is ultra-pure water, and acetic acid is that analysis is pure.
The Kangfuxin Liquid of 10 different batches: lot number 140308, lot number 140309, lot number 140310, lot number 140311, batch
Numbers 140312, lot number 140313, lot number 140314, lot number 140315, lot number 140316, lot number 140317.
Trigonelline, uracil, hypoxanthine, adenosine, inosine (are purchased from Chinese pharmaceutical biological product identification institute, lot number
It is respectively a0654, pcs3048, pcs3055, pcs0217, pcs3058).
(2) chromatographic condition
A. the selection of Detection wavelength
Because Kangfuxin Liquid is the alcohol extracting thing of pharmaceutical insects periplaneta americana, wherein contains a large amount of nucleosides materials, and show
There are some researches show, nucleoside compound (nucleoside, base and the like) is that biological cell sustains life the basic composition of activity
Element, participates in dna metabolic process, has the pharmacologically actives such as anticancer, antiviral.Many animals medicine for example Cordyceps, LUMBRICUS,
All find in Eupolyphaga Seu Steleophaga and snake venom etc. to contain substantial amounts of nucleoside compound, and by as the reference substance setting up finger printing.
Therefore this experimental selection 254nm (nucleosides material has obtained the maximum absorption) is as operation wavelength, the chromatograph according to gradient elution afterwards
Figure, changes reference wavelength it was also found that containing most chromatographic peaks, informative at 254nm.
B. the selection of mobile phase
Acetonitrile and methanol are the most frequently used two kinds of mobile phases in high performance liquid chromatography detection.With 2005 editions " Chinese Pharmacopoeias " it is
Example, wherein with methanol for the medicine of mobile phase main component be 159 kinds, with acetonitrile for the medicine of mobile phase main component be 116
Kind.In chromatographic system, the impact to reversed phase chromatography separation for the strength of mobile phase is very big, if mobile phase is too strong, k value diminishes, and separates
Degree would generally reduce, and solvent strength is too weak, then run time is long, can lose time in the case that batch is many.Acetonitrile is in uv
The noise producing during detection is little, and eluting power is strong, generally more than one times of methanol-eluted fractions ability；But the appearance time of chromatographic peak is past
Toward early than methanol, this often causes to be difficult between chromatographic peak separately, to cause bimodal feelings for the Chinese medicine of complicated component
Condition.From the point of view of environmental protection, the toxicity of methanol is far smaller than acetonitrile.
This experiment compares acetonitrile-water and the chromatogram of methanol-water flow phase system, finds that acetonitrile polarity is bigger than methanol,
Lead to peak figure appearance too fast, and the post pressure of methanol-water system gradient elution is again smaller than acetonitrile-water system.And due to ucleosides
Material assumes alkalescence, therefore methanol -0.1% acetate system eluting is better than methanol-water system elutions effect, and separating effect is preferable,
Retention time is moderate, the precision of method, stability, reproducible.
C. the selection of elution requirement
Because the single Western medicine of Chinese medicine and effective ingredient is different, its complicated component, tend not to take into account using isocratic elution
Some properties differ the separation requirement of larger component it is impossible to ensureing all components all appearances and obtaining suitable separation.Now
Need using the type of elution changing mobile phase concentration proportioning by certain procedures, i.e. gradient elution.The gradient that this experiment is chosen is washed
De-, with methanol -0.1% acetic acid as mobile phase.
D. the selection of other parameters
In reversed-phase liquid chromatography, the range of application of c18 chromatographic column is the widest.This experimental selection hypersil gold
C18 post (4.6 × 150mm, 5 μm), kromasil c18 post (4.6 × 250mm, 5 μm) and inertsustain c18 post (4.6
× 250mm, 5 μm) it is compared, according to the separation degree of chromatogram, final choice inertsustain c18 post (4.6 ×
250mm, 5 μm).And internal diameter is the chromatographic column of 4.6mm, its optimum flow rate is 1.0ml/min, because Chinese medicine ingredients are complicated, in order to
So that chromatographic peak is preferably separated, flow rate set is 0.8ml/min.And the rising with column temperature, chromatographic peak also becomes therewith
Point becomes symmetrical, therefore the column temperature selecting is 35 DEG C.
In sum, the chromatographic condition of the hplc finger printing of Kangfuxin Liquid preparation is as follows: tsq quantum ultra liquid
Phase chromatograph-tandem mass spectrum combined instrument, chromatographic column is hypersil gold c18 post (4.6 × 150mm, 5 μm), and mobile phase is
0.1% acetic acid aqueous solution (a)-methanol (b) gradient elution (condition is as shown in table 1), analysis time 45min, flow velocity 0.8ml/
Min, Detection wavelength 254nm, 35 DEG C of column temperature, sample size 20 μ l.
Table 1 chromatographic elution conditions
(3) linear relationship is investigated
Accurate measuring trigonelline, uracil, hypoxanthine, inosine, the reference substance of adenosine are diluted to concentration for 1 μ respectively
G/ml, 5 μ g/ml, 10 μ g/ml, 20 μ g/ml and 50 μ g/ml, draw 10 μ l injection high performance liquid chromatographs respectively, record chromatograph
Figure, measures its peak area, and with peak area as vertical coordinate (y), sample size (μ g/ml) is abscissa (x), carries out linear regression,
Obtain regression equation as follows: y1=3273.2+57362.2x1 (r1=0.9999), y2=23228.6+173761x2 (r2
=1.0000), y3=39313.1+258077x3 (r3=0.9999), y4=10349.8+143206x4 (r4=0.9999),
Y5=37328.8+291140x5 (r5=0.9999).Result shows, trigonelline, uracil, hypoxanthine, inosine and adenosine
It is in good linear relationship in 1-50ug/ml scope.
(4) sample size measures
Take the Kangfuxin Liquid of 10 batches, prepare solution according to experimental technique, accurately draw reference substance solution and sample is molten
The each 10ul of liquid, is injected separately into high performance liquid chromatograph, and sample size the results are shown in Table 2.From Table 2, it can be seen that this five kinds of samples
Content is relatively stable.
The content of five kinds of comparison quality in table 2 10 batch Kangfuxin Liquid
(5) finger printing
Relevant parameter according to given by 10 batches of Kangfuxin Liquid preparation hplc collection of illustrative plates, aforementioned preparation side pressed by Kangfuxin Liquid preparation
The main chromatographic peak that method is measured gained occurred in 35 minutes；The need testing solution chromatogram of 45 minutes show 35 minutes with
Chromatographic peak does not occur afterwards.The chromatograph graph discovery of relatively each batch sample have 12 peaks be each batch total, by this Criterion fingerprint
Collection of illustrative plates.
(6) have the demarcation of fingerprint peakses
To 10 batches of Kangfuxin Liquid preparation test sample fingerprint map analyzings, using hypoxanthine (No. 6 peaks) as interior reference peak, with
The total rate at peak is foundation more than 80%, has demarcated 12 peaks, respectively No. 1 peak (0.432), No. 2 peaks (0.671), No. 3 peaks
(0.731), No. 4 peaks (0.820), No. 5 peaks (0.888), No. 6 peaks (1.000), No. 7 peaks (1.126), No. 8 peaks (1.462), No. 9
Peak (1.536), No. 10 peaks (1.884), No. 11 peaks (2.282), No. 12 peaks (2.675).By comparing with reference substance collection of illustrative plates, point out
No. 1 peak is trigonelline, and No. 3 peaks are uracil, and No. 6 peaks are hypoxanthine, and No. 8 peaks are inosine, and No. 10 peaks are adenosine.Made with this
The foundation demarcated for total fingerprint peakses is it is allowed to relative deviation is ± 10%.
(7) have fingerprint peakses relative retention time
In 10 batches of Kangfuxin Liquid preparation test sample finger printing, the relative retention time of total fingerprint peakses is shown in Table 3.
Table 3 Kangfuxin Liquid preparation has fingerprint peakses relative retention time (n=10)
The rsd of the relative retention time of 10 batches of test samples is respectively less than 2.0%, shows that the concordance of each batch sample is preferable.
(8) have fingerprint peakses relative peak area
In 10 batches of Kangfuxin Liquid preparation test sample finger printing, the relative peak area of total fingerprint peakses is shown in Table 4.12 have
The rsd value of peak relative peak area is 3.83%-9.27%, shows that the relative amount at each total peak is variant；Although being significantly greater than
The rsd value of relative peak area, but still conform to the requirement to Chinese medicine preparation hplc finger printing for the Chinese Pharmacopoeia.
Table 4 Kangfuxin Liquid preparation has fingerprint peakses relative peak area (n=10)
(9) non-shared peak area
Remove the peak retaining without post, the non-shared peak gross area calculating 10 batches of test samples accounts for the ratio of total peak area,
The results are shown in Table 5.The ratio that each batch of Kangfuxin Liquid preparation non-shared peak gross area accounts for the gross area is respectively less than 10%, meets that " Chinese medicine is noted
Penetrate agent finger printing research technical requirements (provisional) " regulation.
The non-shared peak gross area of table 5 Kangfuxin Liquid preparation accounts for the percentage ratio (n=10) of total peak area
| Lot number | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| Proportion | 7.86 | 6.88 | 7.45 | 7.52 | 5.87 | 5.81 | 6.43 | 6.77 | 7.94 | 6.75 |
(10) Kangfuxin Liquid preparation finger similarity evaluation
Import by the efficient liquid phase initial data of 10 batch Kangfuxin Liquids and by the integration data of area normalization method
Zhejiang University's version Chinese medicine fingerprint Similarity Measure software.Calculated similar between 10 batch Kangfuxin Liquids using Cosin method
Degree, result shows that 10 batch Kangfuxin Liquid similarities are all higher, and minima is 0.98.
(11) Precision Experiment
Kangfuxin Liquid preparation need testing solution of the present invention is taken to be measure object, under chromatographic condition of the present invention, even
Continue and measure 6 times, record relative retention time and the relative peak area of each total chromatographic peak, the results are shown in Table 6～7.Result shows, respectively
The rsd of the relative retention time of chromatographic peak and peak area is respectively less than 2%.Prove that the precision of instrument is good, meet hplc fingerprint
Graphical spectrum technology requires.
Table 6 Kangfuxin Liquid precision test relative retention time
Table 7 Kangfuxin Liquid precision test relative peak area
(12) repeatability experiment
Take 6 parts of Kangfuxin Liquid preparation test sample of the present invention, be analyzed under chromatographic condition of the present invention, record is related
Data.With the hypoxanthic retention time of object of reference as reference, converse the relative retention time at each total peak and peak area
Ratio, the results are shown in Table 8～9.Result shows, the relative retention time of each chromatographic peak and the rsd of relative peak area are respectively less than 3%,
Show that the repeatability of the method is good, meet fingerprint pattern technology requirement.
Table 8 Kangfuxin Liquid reappearance test relative retention time
Table 9 Kangfuxin Liquid reappearance test relative peak area
(13) stability experiment
Kangfuxin Liquid preparation need testing solution of the present invention is taken to be measure object, under chromatographic condition of the present invention, point
Not 0,2,4,6,8,12h detected, record related data.With the hypoxanthic retention time of object of reference as reference, conversion
Go out the relative retention time at each total peak and the ratio of peak area, the results are shown in Table 10～11.Result shows, each chromatographic peak relative
The rsd of retention time and relative peak area is respectively less than 3% it was demonstrated that sample is stable in 12h, and meeting hplc fingerprint pattern technology will
Ask.
Table 10 Kangfuxin Liquid stability test relative retention time
Table 11 Kangfuxin Liquid stability test relative peak area
Embodiment 2
The measuring method of Kangfuxin Liquid preparation finger, comprises the steps:
The preparation of (a) need testing solution: take Kangfuxin Liquid preparation, cross 0.22 μm of microporous filter membrane, take subsequent filtrate, standby；
The preparation of (b) reference substance solution: precision weighs trigonelline, uracil, hypoxanthine, inosine, appropriate adenosine, plus
Water is made every 1ml and is contained trigonelline, uracil, hypoxanthine, inosine and adenosine respectively 20 μ g, 30 μ g, 20 μ g, 40 μ g and 40
The mixing contrast solution of μ g, standby；
C () measures: draw test sample 20 μ l injection high performance liquid chromatographs each with reference substance solution, with 0.08% acetic acid water
Solution (a)-methanol (b) is mobile phase, using gradient elution, obtains the finger printing of Kangfuxin Liquid preparation.
High performance liquid chromatograph condition determination is: filler is octadecylsilane chemically bonded silica；0.08% acetic acid is water-soluble
Liquid (a)-methanol (b)；Using gradient elution: 0～3min, 3%b；3～5min, 3%b～8%b；5～10min, 8%b～
10%b；10～15min, 10%b～35%b；15～30min, 35%b～80%b；30～40min, 80%b～3%b；40
～45min, 3%b；Detection wavelength is 254nm；Column temperature is 35 DEG C；Flow velocity is 0.8ml/min；Number of theoretical plate is based on hypoxanthine
Calculation is not less than 5000.
The Kangfuxin Liquid preparation finger obtaining has 12 total fingerprint peakses, wherein 5 respectively with corresponding reference
Thing peak retention time is identical；Peak corresponding with hypoxanthine object of reference peak is s peak, the relative retention time of 12 total fingerprint peakses
It is respectively as follows: No. 1 peak 0.432,2 peak 0.673,3 peak 0.735,4 peak 0.821,5 peak 0.888,6 (s) number peak 1.000,7
Number peak 1.127,8 peak 1.460,9 peak 1.537,10 peak 1.886,11 peak 2.284,12 peak 2.674.
Embodiment 3
The measuring method of Kangfuxin Liquid preparation finger, comprises the steps:
The preparation of (a) need testing solution: take Kangfuxin Liquid preparation, cross 0.22 μm of microporous filter membrane, take subsequent filtrate, standby；
The preparation of (b) reference substance solution: precision weighs trigonelline, uracil, hypoxanthine, inosine, appropriate adenosine, plus
Water is made every 1ml and is contained trigonelline, uracil, hypoxanthine, inosine and adenosine respectively 20 μ g, 30 μ g, 20 μ g, 40 μ g and 40
The mixing contrast solution of μ g, standby；
C () measures: draw test sample 20 μ l injection high performance liquid chromatographs each with reference substance solution, with 1.0% acetic acid water
Solution (a)-methanol (b) is mobile phase, using gradient elution, obtains the finger printing of Kangfuxin Liquid preparation.
High performance liquid chromatograph condition determination is: filler is octadecylsilane chemically bonded silica；1.0% acetic acid aqueous solution
(a)-methanol (b)；Using gradient elution: 0～3min, 3%b；3～5min, 3%b～8%b；5～10min, 8%b～10%
b；10～15min, 10%b～35%b；15～30min, 35%b～80%b；30～40min, 80%b～3%b；40～
45min, 3%b；Detection wavelength is 254nm；Column temperature is 35 DEG C；Flow velocity is 0.8ml/min；Number of theoretical plate is pressed hypoxanthine and is calculated
It is not less than 5000.
The Kangfuxin Liquid preparation finger obtaining has 12 total fingerprint peakses, wherein 5 respectively with corresponding reference
Thing peak retention time is identical；Peak corresponding with hypoxanthine object of reference peak is s peak, the relative retention time of 12 total fingerprint peakses
It is respectively as follows: No. 1 peak 0.432,2 peak 0.670,3 peak 0.731,4 peak 0.820,5 peak 0.888,6 (s) number peak 1.000,7
Number peak 1.125,8 peak 1.466,9 peak 1.537,10 peak 1.882,11 peak 2.282,12 peak 2.677.
Embodiment 4
The measuring method of Kangfuxin Liquid preparation finger, comprises the steps:
The preparation of (a) need testing solution: take Kangfuxin Liquid preparation, cross 0.22 μm of microporous filter membrane, take subsequent filtrate, standby；
The preparation of (b) reference substance solution: precision weighs trigonelline, uracil, hypoxanthine, inosine, appropriate adenosine, plus
Water is made every 1ml and is contained trigonelline, uracil, hypoxanthine, inosine and adenosine respectively 20 μ g, 30 μ g, 20 μ g, 40 μ g and 40
The mixing contrast solution of μ g, standby；
C () measures: draw test sample 20 μ l injection high performance liquid chromatographs each with reference substance solution, with 1.2% acetic acid water
Solution (a)-methanol (b) is mobile phase, using gradient elution, obtains the finger printing of Kangfuxin Liquid preparation.
High performance liquid chromatograph condition determination is: filler is octadecylsilane chemically bonded silica；1.2% acetic acid aqueous solution
(a)-methanol (b)；Using gradient elution: 0～3min, 3%b；3～5min, 3%b～8%b；5～10min, 8%b～10%
b；10～15min, 10%b～35%b；15～30min, 35%b～80%b；30～40min, 80%b～3%b；40～
45min, 3%b；Detection wavelength is 254nm；Column temperature is 35 DEG C；Flow velocity is 0.8ml/min；Number of theoretical plate is pressed hypoxanthine and is calculated
It is not less than 5000.
The Kangfuxin Liquid preparation finger obtaining has 12 total fingerprint peakses, wherein 5 respectively with corresponding reference
Thing peak retention time is identical；Peak corresponding with hypoxanthine object of reference peak is s peak, the relative retention time of 12 total fingerprint peakses
It is respectively as follows: No. 1 peak 0.438,2 peak 0.679,3 peak 0.736,4 peak 0.824,5 peak 0.890,6 (s) number peak 1.000,7
Number peak 1.129,8 peak 1.467,9 peak 1.540,10 peak 1.882,11 peak 2.280,12 peak 2.678.
Claims (4)
1. the measuring method of Kangfuxin Liquid preparation finger is it is characterised in that the method comprises the steps:
The preparation of (a) need testing solution: take Kangfuxin Liquid preparation, cross 0.22 μm of microporous filter membrane, take subsequent filtrate, standby；
The preparation of (b) reference substance solution: precision weighs trigonelline, uracil, hypoxanthine, inosine, appropriate adenosine, the system of adding water
Become every 1ml to contain trigonelline, uracil, hypoxanthine, inosine and adenosine and be respectively 20 μ g, 30 μ g, 20 μ g, 40 μ g and 40 μ g
Mixing contrast solution, standby；
C () finger printing makes: draw each 20 μ l injection high performance liquid chromatographs of test sample and reference substance solution, with 0.08%～
1.2% acetic acid aqueous solution (a)-methanol (b) is mobile phase, using gradient elution, obtains the finger printing of Kangfuxin Liquid preparation.
2. Kangfuxin Liquid preparation according to claim 1 fingerprint pattern quality determination method it is characterised in that: step
C in (), preferred mobile phase is 0.1% acetic acid aqueous solution (a)-methanol (b).
3. the fingerprint pattern quality determination method of Kangfuxin Liquid preparation according to claim 1 is it is characterised in that efficient liquid
Chromatography condition determination is: filler is octadecylsilane chemically bonded silica；0.08%～1.2% acetic acid aqueous solution (a)-first
Alcohol (b)；Using gradient elution: 0～3min, 3%b；3～5min, 3%b～8%b；5～10min, 8%b～10%b；10～
15min, 10%b～35%b；15～30min, 35%b～80%b；30～40min, 80%b～3%b；40～45min, 3%
b；Detection wavelength is 254nm；Column temperature is 35 DEG C；Flow velocity is 0.8ml/min；Number of theoretical plate is calculated by hypoxanthine and is not less than
5000.
4. the finger printing that the fingerprint pattern quality determination method of Kangfuxin Liquid preparation according to claim 1 and 2 obtains,
It is characterized in that: Kangfuxin Liquid preparation has 12 total fingerprint peakses, wherein 5 respectively with corresponding object of reference peak retention time
Identical；Peak corresponding with hypoxanthine object of reference peak is s peak, and the relative retention time of 12 total fingerprint peakses is respectively as follows: No. 1 peak
0.432 ± 0.043, No. 2 peaks 0.671 ± 0.067,3 peak 0.731 ± 0.073,4 peak 0.820 ± 0.082,5 peak 0.888
± 0.089,6 (s) number peak 1.000 ± 0.000,7 peak 1.126 ± 0.113,8 peak 1.462 ± 0.146,9 peak 1.536 ±
0.154, No. 10 peak 1.884 ± 0.118,11 peak 2.282 ± 0.228,12 peak 2.675 ± 0.268.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107490614A (en) * | 2017-09-15 | 2017-12-19 | 福建中医药大学 | A kind of quality determining method of Kangfuxin Liquid |
| CN107729721A (en) * | 2017-10-17 | 2018-02-23 | 中国科学院上海有机化学研究所 | A kind of metabolin identification and disorderly path analysis method |
| CN109324141A (en) * | 2018-12-18 | 2019-02-12 | 广州市药材公司中药饮片厂 | The characteristic fingerprint pattern and its construction method of a kind of wide dragon or its processed product and application |
| CN109959732A (en) * | 2017-12-26 | 2019-07-02 | 内蒙古京新药业有限公司 | The separation method of Kangfuxin Liquid finger-print and its effective constituents A |
| CN113607855A (en) * | 2021-08-23 | 2021-11-05 | 神威药业集团有限公司 | Fingerprint spectrum of medicinal preparation of snakegourd fruit, allium macrostemon and pinellia ternata decoction, and establishing method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101315355A (en) * | 2008-07-17 | 2008-12-03 | 四川科伦药业股份有限公司 | Fingerprint pattern quality determination method for novel healing formulation |
| CN101320026A (en) * | 2008-07-17 | 2008-12-10 | 四川科伦药业股份有限公司 | Quality detection method of novel healing formulation |
| 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 |
-
2016
- 2016-08-18 CN CN201610688619.5A patent/CN106370739A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101315355A (en) * | 2008-07-17 | 2008-12-03 | 四川科伦药业股份有限公司 | Fingerprint pattern quality determination method for novel healing formulation |
| CN101320026A (en) * | 2008-07-17 | 2008-12-10 | 四川科伦药业股份有限公司 | Quality detection method of novel healing formulation |
| 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 |
Non-Patent Citations (4)
| Title |
|---|
| 吴红梅: "上市品种 "康复新液"的药学再评价", 《中国博士学位论文全文数据库 医药卫生特辑》 * |
| 吴红梅等: "康复新液 UPLC 指纹图谱研究", 《中国实验方剂学杂志》 * |
| 张丹: "美洲大蠊抗肿瘤活性成分及其结肠定位片的研究", <中国博士学位论文全文数据库 医药卫生特辑> * |
| 张利等: "HPLC 法同时测定康复新液中尿嘧啶、次黄嘌呤及肌苷含量", 《中国药师》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107490614A (en) * | 2017-09-15 | 2017-12-19 | 福建中医药大学 | A kind of quality determining method of Kangfuxin Liquid |
| CN107490614B (en) * | 2017-09-15 | 2019-11-26 | 福建中医药大学 | A kind of quality determining method of Kangfuxin Liquid |
| CN107729721A (en) * | 2017-10-17 | 2018-02-23 | 中国科学院上海有机化学研究所 | A kind of metabolin identification and disorderly path analysis method |
| CN107729721B (en) * | 2017-10-17 | 2021-01-19 | 中国科学院上海有机化学研究所 | Metabolite identification and disorder pathway analysis method |
| CN109959732A (en) * | 2017-12-26 | 2019-07-02 | 内蒙古京新药业有限公司 | The separation method of Kangfuxin Liquid finger-print and its effective constituents A |
| CN109324141A (en) * | 2018-12-18 | 2019-02-12 | 广州市药材公司中药饮片厂 | The characteristic fingerprint pattern and its construction method of a kind of wide dragon or its processed product and application |
| CN109324141B (en) * | 2018-12-18 | 2022-05-03 | 广州白云山中药饮片有限公司 | Characteristic fingerprint spectrum of guangdong or processed products thereof, and construction method and application thereof |
| CN113607855A (en) * | 2021-08-23 | 2021-11-05 | 神威药业集团有限公司 | Fingerprint spectrum of medicinal preparation of snakegourd fruit, allium macrostemon and pinellia ternata decoction, and establishing method and application thereof |
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