CN107064320B - A kind of method of quality control and its finger-print of reinforcing spleen and kidney side - Google Patents
A kind of method of quality control and its finger-print of reinforcing spleen and kidney side Download PDFInfo
- Publication number
- CN107064320B CN107064320B CN201610917155.0A CN201610917155A CN107064320B CN 107064320 B CN107064320 B CN 107064320B CN 201610917155 A CN201610917155 A CN 201610917155A CN 107064320 B CN107064320 B CN 107064320B
- Authority
- CN
- China
- Prior art keywords
- rsd
- reinforcing spleen
- kidney side
- mobile phase
- peak area
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- 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
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of method of quality control of reinforcing spleen and kidney side and its finger-prints.The method of quality control comprises the following steps: S1. prepares reinforcing spleen and kidney side's test solution；S2. it draws test solution and injects liquid chromatograph, record chromatogram；S3. chromatogram step S2 obtained imports similarity evaluation；Select in the chromatogram of different batches reinforcing spleen and kidney side existing chromatographic peak as shared peak；The standard finger-print of reinforcing spleen and kidney side is generated with mean value calculation method；Calculate relative retention time, the relative peak area at each shared peak；S4. the finger-print of reinforcing spleen and kidney side to be detected is established；Then it is compared with standard finger-print described in step S3, finally judges whether reinforcing spleen and kidney side's quality to be detected is qualified.The method avoids the conventional one-sidedness for only measuring one, two chemical component and determining reinforcing spleen and kidney side's quality, is conducive to comprehensive control to Chinese medicine compound prescription quality.
Description
Technical field
The present invention relates to Chinese medicine detection technique fields, and in particular to a kind of method of quality control of reinforcing spleen and kidney side and its refers to
Line map.
Background technique
Chronic kidney disease Chronic kidney disease (CKD) refer to kidney injury or glomerular filtration rate (GFR) <
60mL/(min·1.73m2) continue 3 months or more；The definition of injury of kidney refer to Renal Paphology exception or blood, urine,
The inspection of iconography is abnormal.CKD is the general name of various primary and secondary kidney trouble, is the chronic non infectious of one group of carry out sexual development
Property disease, China illness rate in rising year by year trend, become the serious challenge that whole world country must face.CKD is from kidney
Function was normally divided into for 5 phases to the whole process of renal failure, chronic nephritis and 2 stages of chronic kidney hypofunction was equivalent to, wherein slowly
The property kidney trouble fourth phase is the diagnosis for meeting chronic kidney disease.
Reinforcing spleen and kidney side is Guangdong Provincial TCM Hospital Nephrology dept. Liu Xusheng professor on the Research foundation of this field, integrates industry
Essence simultaneously carries out repeatedly demonstration and forms (a kind of Chinese materia medica preparation and preparation method thereof for treating chronic kidney disease, patent after optimizing
Publication number: CN102579894A).The prescription includes nine herbal medicines such as Radix Astragali, raw Radix Codonopsis, Semen Cuscutae, the function with reinforcing spleen and kidney
Effect, it is significant in efficacy to chronic kidney disease, clinical efficacy show the group can delaying chronic kidney popular name for fourth phase patient eGFR decline,
The progress for having delayed this disease, better than the active drug ACEI used in the world.
Tcm clinical efficacy, cannot widely available international community for a long time due to lacking reliable Clinical Evidence support
Approval.Using the standard scheme-- randomized double-blind clinical research test of evaluation intervening measure effect generally acknowledged in the world
(Randomized Controlled trials, RTC), the clinical experience that traditional Chinese medicine is treated is with the evidence form of scientific research
It shows, is a traditional Chinese medicine break-through point in line with international standards.And Chinese medicine compound prescription quality control standard is the control of RTC mass
Source.Therefore, tcm clinical curative effect is intended to by international endorsement, should there is standardized traditional Chinese medicine quality control standard first.
The mass action feature that Chinese medicine compound prescription has cannot be only for certain several chemical component, therefore should establish on the whole
The method of concentrated expression traditional Chinese medicine quality, using finger-print as Chinese patent drug, this kind of matter containing compounding substances group of herb extracts
Amount control method has become current international consensus.Traditional Chinese medicine fingerprint is to Chinese medicine and its compound material base global feature
A kind of quantitative expression and parsing means, after referring to that certain Chinese medicine or Chinese patent drug are appropriately processed, using certain analysis hand
Section extracts its chemical information, obtains the map at the shared peak that can indicate its characteristic.The guarantee of Chinese medicine and its compound inherent quality
It also is the basic requirement of the modernization of Chinese medicine, Chinese medicine compound prescription complicated component, the effective component overwhelming majority is not clear, needs in foundation
Traditional Chinese medicine quality evaluation criteria.Reinforcing spleen and kidney side is that the Chinese medicine including nine herbal medicines such as Radix Astragali, raw Radix Codonopsis, Semen Cuscutae is multiple
Side, clinically for treating chronic kidney disease, curative effect has obtained clinical verifying, and whether can guarantee the quality of drug
It is the basis for determining reinforcing spleen and kidney side's clinical efficacy.If only illustrating the inherent quality tool of prescription with one, two kinds of active components
There is certain one-sidedness, and cannot be guaranteed whether selected index components are active.
Summary of the invention
The technical problem to be solved by the present invention is to can control reinforcing spleen and kidney side inherence to overcome to lack in the prior art
The method of quality control of quality provides a kind of method of quality control of reinforcing spleen and kidney side.This method can measure tonifying spleen benefit simultaneously
In kidney side in the multi-flavors medicine such as Radix Astragali, Radix Codonopsis, Radix Salviae Miltiorrhizae, prepared fleece flower root, Semen Cuscutae multiclass active constituent finger print measuring method,
By this measuring method, the inherent quality of reinforcing spleen and kidney side can control.
Above-mentioned technical problem to be solved by this invention, is achieved by the following technical programs:
A kind of method of quality control of reinforcing spleen and kidney side, comprises the following steps:
S1. reinforcing spleen and kidney side's test solution is prepared；
S2. it draws test solution and injects liquid chromatograph, record chromatogram；
S3. chromatogram step S2 obtained imports similarity evaluation；Selection different batches
Existing chromatographic peak is as shared peak in the chromatogram of secondary reinforcing spleen and kidney side；Reinforcing spleen and kidney side is generated with mean value calculation method
Standard finger-print；Calculate relative retention time, the relative peak area at each shared peak；
S4. the finger-print of reinforcing spleen and kidney side to be detected is established；Then it is carried out with standard finger-print described in step S3
Compare, finally judges whether reinforcing spleen and kidney side's quality to be detected is qualified.
The drying that inventor is prepared for reinforcing spleen and kidney side first is sprayed powder, by the preparation method to prescription,
The selection and chromatographic condition methodological study of chromatographic condition, establish reinforcing spleen and kidney side's finger-print.The finger-print can meet
It detects and analyzes while multiclass active constituent in nine taste medicine of reinforcing spleen and kidney side, and this finger-print is used for the quality of the prescription
Control.
Preferably, test solution described in step S1 is prepared via a method which to obtain:
S11. Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Poria cocos, Chinese yam, the fleece-flower root, nine taste of Semen Cuscutae, semen coicis and Radix Salviae Miltiorrhizae of crushing are taken
Chinese medicine, after mixing plus water, 20~50min of immersion are decocted 0.5~3 hour, after filtering again after boiling, and filtrate is recovered under reduced pressure, spray
Mist is dry, obtains dried powder；
S12. weigh reinforcing spleen and kidney side's dried powder, be dissolved in acetonitrile solution or methanol aqueous solution, ultrasonic dissolution, constant volume in
Volumetric flask, filter membrane are spare.
Preferably, the concrete operations of step S11 are as follows: take 15~20 parts of 25~33 parts of Radix Astragali, the Radix Codonopsis of crushing, Rhizoma Atractylodis Macrocephalae 10~
18 parts, 15~20 parts of Poria cocos, 13~20 parts of Chinese yam, 15~20 parts of the fleece-flower root, 12~20 parts of Semen Cuscutae, 15~25 parts of semen coicis,
10~20 parts of Radix Salviae Miltiorrhizae, after mixing plus water, immersion 30min are decocted 2 hours, after filtering, filter residue decocts one with method again again after boiling
It is secondary, merge filtrate twice, be recovered under reduced pressure, is spray-dried, obtains dried powder.
Most preferably, the concrete operations of step S12 are as follows: weigh reinforcing spleen and kidney side dried powder 56mg, be dissolved in 2mL 60%
Acetonitrile solution, ultrasonic dissolution, constant volume crosses 0.22 μm of filter membrane in 5mL volumetric flask, spare.
It is highly preferred that in step S2 liquid chromatograph chromatographic condition are as follows: chromatographic column is with octadecylsilane chemically bonded silica
For filler；Mobile phase A is 0.1%-0.5% aqueous formic acid, and Mobile phase B is acetonitrile, using gradient elution；Column temperature is 25~35
℃；Flow velocity is 200~400 μ L/min；Sample volume is 1~10 μ L.
It is further preferred that the condition of gradient elution are as follows: mobile phase A is 0.1% aqueous formic acid, Mobile phase B
For acetonitrile；At 0 minute, 88% mobile phase A, 12% Mobile phase B；At 22 minutes, 75% mobile phase A, 25% Mobile phase B；35 points
Zhong Shi, 65% mobile phase A, 35% Mobile phase B；At 45 minutes, 50% mobile phase A, 50% Mobile phase B；At 52 minutes, 45% stream
Dynamic phase A, 55% Mobile phase B；Mobile phase A and Mobile phase B configuration proportion are prepared by volume.
Most preferably, column temperature is 25 DEG C；Flow velocity is 250 μ L/min；Sample volume is 2 μ L；Chromatographic column selects Kinetex C18
Analytical column, specification be 2.1mm × 100mm, 1.7 μm.
Preferably, it is also connected with mass spectrograph after the liquid chromatograph described in step S2, for detecting the molecule at shared peak
Amount；The mass spectrograph use electric spray ion source (ESI), sheath gas be 45 units, auxiliary gas be 6 units, spray voltage for-
2.5~-3.5kV, capillary temperature are 300~375 DEG C, and sample first uses high-resolution to carry out full scan, and second order ms are using dynamic
The scanning of state data dependency, collision energy are set as 20~45%.
A kind of finger-print of reinforcing spleen and kidney side, is established to obtain by above-mentioned method for building up, includes shared peak as follows:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%.
Preferably, the finger-print includes following characteristic peak:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 5 peaks, Average residence time RT are 15.62min, and RSD 1.48%, peak area 5049.76, RSD is
3.72%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 14 peaks, Average residence time RT are 22.66min, and RSD 3.39%, peak area 3415.56, RSD is
24.4%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 17 peaks, Average residence time RT are 30.95min, and RSD 1.63%, peak area 4498.02, RSD is
5.54%；
No. 18 peaks, Average residence time RT are 31.85min, and RSD 1.89%, peak area 4870.13, RSD is
2.16%；
No. 21 peaks, Average residence time RT are 34.14min, and RSD 1.90%, peak area 4582.24, RSD is
17.46%；
No. 23 peaks, Average residence time RT are 35.08min, and RSD 1.94%, peak area 3040.11, RSD is
8.13%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%；
No. 27 peaks, Average residence time RT are 41.28min, and RSD 2.03%, peak area 5646.33, RSD is
2.16%；
No. 28 peaks, Average residence time RT are 43.07min, and RSD 2.23%, peak area 3629.00, RSD is
13.16%.
Most preferably, the finger-print includes following characteristic peak:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 2 peaks, Average residence time RT are 11.28min, and RSD 0.52%, peak area 1095.05, RSD is
3.49%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 4 peaks, Average residence time RT are 15.17min, and RSD 2.80%, peak area 1158.83, RSD is
15.83%；
No. 5 peaks, Average residence time RT are 15.62min, and RSD 1.48%, peak area 5049.76, RSD is
3.72%；
No. 6 peaks, Average residence time RT are 16.28min, and RSD 2.11%, peak area 582.55, RSD is
16.58%；
No. 7 peaks, Average residence time RT are 17.09min, and RSD 3.44%, peak area 1532.83, RSD is
18.40%；
No. 8 peaks, Average residence time RT are 18.72min, and RSD 3.12%, peak area 2777.4, RSD is
3.27%；
No. 9 peaks, Average residence time RT are 19.45min, and RSD 3.32%, peak area 941.96, RSD is
17.72%；
No. 10 peaks, Average residence time RT are 20.99min, and RSD 1.35%, peak area 2708.15, RSD is
1.38%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 13 peaks, Average residence time RT are 22.81min, and RSD 1.04%, peak area 1122.84, RSD is
18.86%；
No. 14 peaks, Average residence time RT are 22.66min, and RSD 3.39%, peak area 3415.56, RSD is
24.4%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 16 peaks, Average residence time RT are 25.13, RSD 1.56%, and peak area 2253.63, RSD is
16.97%；
No. 17 peaks, Average residence time RT are 30.95min, and RSD 1.63%, peak area 4498.02, RSD is
5.54%；
No. 18 peaks, Average residence time RT are 31.85min, and RSD 1.89%, peak area 4870.13, RSD is
2.16%；
No. 19 peaks, Average residence time RT are 32.82min, and RSD 1.88%, peak area 1216.68, RSD is
13.19%；
No. 20 peaks, Average residence time RT are 33.96min, and RSD 1.89%, peak area 1549.99, RSD is
10.15%；
No. 21 peaks, Average residence time RT are 34.14min, and RSD 1.90%, peak area 4582.24, RSD is
17.46%；
No. 22 peaks, Average residence time RT are 34.48min, and RSD 1.96%, peak area 1521.18, RSD is
3.76%；
No. 23 peaks, Average residence time RT are 35.08min, and RSD 1.94%, peak area 3040.11, RSD is
8.13%；
No. 24 peaks, Average residence time RT are 35.35min, and RSD 1.90%, peak area 2944.64, RSD is
2.12%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%；
No. 26 peaks, Average residence time RT are 37.11, RSD 1.67%, peak area 718.58, RSD 8.92%；
No. 27 peaks, Average residence time RT are 41.28min, and RSD 2.03%, peak area 5646.33, RSD is
2.16%；
No. 28 peaks, Average residence time RT are 43.07min, and RSD 2.23%, peak area 3629.00, RSD is
13.16%；
No. 29 peaks, Average residence time RT are 43.30min, and RSD 1.96%, peak area 2900.10, RSD is
5.12%.
The utility model has the advantages that (1) present invention using main component in reinforcing spleen and kidney side as Index Establishment chromatographic fingerprinting, realizes it
The chemical component of maximum possible detects, and avoids conventional only one, two chemical component of measurement and determines reinforcing spleen and kidney side's quality
One-sidedness.The chromatographic fingerprinting represents reinforcing spleen and kidney side's major part pharmacological activity, can Efficient Characterization reinforcing spleen and kidney prescription
Quality, be conducive to comprehensive control to Chinese medicine compound prescription quality.(2) present invention provides a kind of detection side of reinforcing spleen and kidney prescription
Method, experiment is practical, has repeatability.Method is stable, easy, precision is high, is easy to grasp.It can be suitable for tonifying spleen benefit simultaneously
The determining fingerprint pattern of kidney side's extracting solution and the determining fingerprint pattern of reinforcing spleen and kidney square preparation.
Detailed description of the invention
Fig. 1 is reinforcing spleen and kidney side's finger-print of the present invention.
Specific embodiment
The present invention is explained further below in conjunction with specific embodiment, but embodiment does not do any type of limit to the present invention
It is fixed.
The person that is not specified actual conditions in embodiment, carries out according to conventional conditions or manufacturer's recommended conditions.Wherein, used
Production firm person is not specified in reagent or instrument, and being can be with conventional products that are commercially available.
Embodiment 1
A kind of method of quality control of reinforcing spleen and kidney side, comprises the following steps:
S1. reinforcing spleen and kidney side's test solution is prepared；
S2. it draws test solution and injects liquid chromatograph, record chromatogram；
S3. chromatogram step S2 obtained imports similarity evaluation；Selection different batches
Existing chromatographic peak is as shared peak in the chromatogram of secondary reinforcing spleen and kidney side；Reinforcing spleen and kidney side is generated with mean value calculation method
Standard finger-print；Calculate relative retention time, the relative peak area at each shared peak；
S4. the finger-print of reinforcing spleen and kidney side to be detected is established；Then it is carried out with standard finger-print described in step S3
Compare, finally judges whether reinforcing spleen and kidney side's quality to be detected is qualified；
Test solution described in step S1 is prepared via a method which to obtain:
Take 18 parts, Tu of 18 parts of 15 parts of 20 parts of 28 parts of Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Poria cocos, 15 parts of Chinese yam, the fleece-flower root of crushing
Sub 15 parts, 22 parts of semen coicis, 18 parts of Radix Salviae Miltiorrhizae, after mixing plus water, immersion 30min are decocted 2 hours, after filtering, filter residue again after boiling
It is decocted again with method once, merges filtrate twice, be recovered under reduced pressure, be spray-dried, obtain dried powder；
S12. weigh reinforcing spleen and kidney side dried powder 56mg, be dissolved in 60% acetonitrile solution of 2mL, ultrasonic dissolution, constant volume in
5mL volumetric flask crosses 0.22 μm of filter membrane, spare；
The chromatographic condition of liquid chromatograph in step S2 are as follows: chromatographic column is using octadecylsilane chemically bonded silica as filler；Stream
Dynamic phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile, using gradient elution；Column temperature is 25 DEG C；Flow velocity is 250 μ L/min；
Sample volume is 2 μ L；Chromatographic column selects Kinetex C18Analytical column, specification be 2.1mm × 100mm, 1.7 μm；
The condition of gradient elution are as follows: mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile；At 0 minute,
88% mobile phase A, 12% Mobile phase B；At 22 minutes, 75% mobile phase A, 25% Mobile phase B；At 35 minutes, 65% mobile phase
A, 35% Mobile phase B；At 45 minutes, 50% mobile phase A, 50% Mobile phase B；At 52 minutes, 45% mobile phase A, 55% flowing
Phase B；Mobile phase A and Mobile phase B configuration proportion are prepared by volume.
The finger-print that the method obtains through this embodiment carries out finger-print to total 10 batches of reinforcing spleen and kidney sides
Analysis, similarity is 90% or more.Wherein unimodal relative response is more than that 2% shared peak has 29, is respectively as follows:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 2 peaks, Average residence time RT are 11.28min, and RSD 0.52%, peak area 1095.05, RSD is
3.49%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 4 peaks, Average residence time RT are 15.17min, and RSD 2.80%, peak area 1158.83, RSD is
15.83%；
No. 5 peaks, Average residence time RT are 15.62min, and RSD 1.48%, peak area 5049.76, RSD is
3.72%；
No. 6 peaks, Average residence time RT are 16.28min, and RSD 2.11%, peak area 582.55, RSD is
16.58%；
No. 7 peaks, Average residence time RT are 17.09min, and RSD 3.44%, peak area 1532.83, RSD is
18.40%；
No. 8 peaks, Average residence time RT are 18.72min, and RSD 3.12%, peak area 2777.4, RSD is
3.27%；
No. 9 peaks, Average residence time RT are 19.45min, and RSD 3.32%, peak area 941.96, RSD is
17.72%；
No. 10 peaks, Average residence time RT are 20.99min, and RSD 1.35%, peak area 2708.15, RSD is
1.38%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 13 peaks, Average residence time RT are 22.81min, and RSD 1.04%, peak area 1122.84, RSD is
18.86%；
No. 14 peaks, Average residence time RT are 22.66min, and RSD 3.39%, peak area 3415.56, RSD is
24.4%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 16 peaks, Average residence time RT are 25.13, RSD 1.56%, and peak area 2253.63, RSD is
16.97%；
No. 17 peaks, Average residence time RT are 30.95min, and RSD 1.63%, peak area 4498.02, RSD is
5.54%；
No. 18 peaks, Average residence time RT are 31.85min, and RSD 1.89%, peak area 4870.13, RSD is
2.16%；
No. 19 peaks, Average residence time RT are 32.82min, and RSD 1.88%, peak area 1216.68, RSD is
13.19%；
No. 20 peaks, Average residence time RT are 33.96min, and RSD 1.89%, peak area 1549.99, RSD is
10.15%；
No. 21 peaks, Average residence time RT are 34.14min, and RSD 1.90%, peak area 4582.24, RSD is
17.46%；
No. 22 peaks, Average residence time RT are 34.48min, and RSD 1.96%, peak area 1521.18, RSD is
3.76%；
No. 23 peaks, Average residence time RT are 35.08min, and RSD 1.94%, peak area 3040.11, RSD is
8.13%；
No. 24 peaks, Average residence time RT are 35.35min, and RSD 1.90%, peak area 2944.64, RSD is
2.12%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%；
No. 26 peaks, Average residence time RT are 37.11, RSD 1.67%, peak area 718.58, RSD 8.92%；
No. 27 peaks, Average residence time RT are 41.28min, and RSD 2.03%, peak area 5646.33, RSD is
2.16%；
No. 28 peaks, Average residence time RT are 43.07min, and RSD 2.23%, peak area 3629.00, RSD is
13.16%；
No. 29 peaks, Average residence time RT are 43.30min, and RSD 1.96%, peak area 2900.10, RSD is
5.12%.
Wherein unimodal relative response is more than that 10% shared peak has 15, is respectively as follows:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 5 peaks, Average residence time RT are 15.62min, and RSD 1.48%, peak area 5049.76, RSD is
3.72%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 14 peaks, Average residence time RT are 22.66min, and RSD 3.39%, peak area 3415.56, RSD is
24.4%；
No. 15 peaks, Average residence time RT are 23.50min, and RSD 1.15%, peak area 3759.06, RSD is
8.75%；
No. 17 peaks, Average residence time RT are 30.95min, and RSD 1.63%, peak area 4498.02, RSD is
5.54%；
No. 18 peaks, Average residence time RT are 31.85min, and RSD 1.89%, peak area 4870.13, RSD is
2.16%；
No. 21 peaks, Average residence time RT are 34.14min, and RSD 1.90%, peak area 4582.24, RSD is
17.46%；
No. 23 peaks, Average residence time RT are 35.08min, and RSD 1.94%, peak area 3040.11, RSD is
8.13%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%；
No. 27 peaks, Average residence time RT are 41.28min, and RSD 2.03%, peak area 5646.33, RSD is
2.16%；
No. 28 peaks, Average residence time RT are 43.07min, and RSD 2.23%, peak area 3629.00, RSD is
13.16%；
Wherein unimodal relative response is more than that 20% shared peak has 5, is respectively as follows:
No. 1 peak, Average residence time RT are 10.87min, and RSD 2.31%, peak area 6049.49, RSD is
2.74%；
No. 3 peaks, Average residence time RT are 11.92min, and RSD 1.43%, peak area 15269.97, RSD is
3.56%；
No. 11 peaks, Average residence time RT are 21.42min, and RSD 1.17%, peak area 14171.91, RSD is
18.94%；
No. 12 peaks, Average residence time RT are 22.17min, and RSD 1.17%, peak area 8738.37, RSD is
2.67%；
No. 25 peaks, Average residence time RT are 35.62min, and RSD 1.91%, peak area 9546.48, RSD is
1.93%；
Embodiment 2
Further, it is also connected with mass spectrograph after the liquid chromatograph described in 1 step S2 of embodiment, it is shared for detecting
The molecular weight at peak；The mass spectrograph uses electric spray ion source (ESI), and sheath gas is 45 units, and auxiliary gas is 6 units, spraying
Voltage is -2.5~-3.5kV, and capillary temperature is 300~375 DEG C, and sample first uses high-resolution to carry out full scan, second order ms
It is scanned using dynamic data dependence, collision energy is set as 20~45%.
Further, on the basis of embodiment 1 also use calycosin-7-O-β-D-glycoside, ononin,
Calycosin, Stibene-glucoside, Rosmarinic acid, alkannic acid, salviandic acid A, tanshin polyphenolic acid B, astragalin, lobetyolin, awns handle
Florigen, Astragaloside IV, astragaloside I, astragaloside II etc. are object of reference, accurately weighed appropriate, are dissolved, are made with 20% acetonitrile
Standard solution, sample introduction is analyzed by HPLC (referring to the HPLC condition of embodiment 1), right according to retention time and ultraviolet characteristic absorption
Shared peak is belonged to.
The present embodiment can be further determined by carrying out molecular weight detection and ownership to shared peak by finger-print
Whether reinforcing spleen and kidney side's quality to be detected is qualified.
The ownership at each shared peak is shown in Table 1 in reinforcing spleen and kidney side, and the finger-print retention time at shared peak, peak area are averaged
The molecular weight of value, RSD value and each shared peak is shown in Table 2.
The ownership table at each shared peak in 1. reinforcing spleen and kidney side of table
Table 2. shares the finger-print retention time at peak, the average value of peak area, RSD value and molecular weight form
Claims (6)
1. a kind of method of quality control of reinforcing spleen and kidney side, which is characterized in that comprise the following steps:
S1. reinforcing spleen and kidney side's test solution is prepared；
S2. it draws test solution and injects liquid chromatograph, record chromatogram；
S3. chromatogram step S2 obtained imports similarity evaluation；Different batches are selected to mend
Existing chromatographic peak is as shared peak in the chromatogram of spleen and tonifying kidney side；The standard of reinforcing spleen and kidney side is generated with mean value calculation method
Finger-print；Calculate relative retention time, the relative peak area at each shared peak；
S4. the finger-print of reinforcing spleen and kidney side to be detected is established；Then it is compared with standard finger-print described in step S3,
Finally judge whether reinforcing spleen and kidney side's quality to be detected is qualified；
The chromatographic condition of liquid chromatograph in step S2 are as follows: chromatographic column is using octadecylsilane chemically bonded silica as filler；Mobile phase A
For 0.1% aqueous formic acid, Mobile phase B is acetonitrile, using gradient elution；Column temperature is 25~35 DEG C；Flow velocity is 200~400 μ L/
min；Sample volume is 1~10 μ L；The condition of gradient elution are as follows:
At 0 minute, 88% mobile phase A, 12% Mobile phase B；At 22 minutes, 75% mobile phase A, 25% Mobile phase B；35 minutes
When, 65% mobile phase A, 35% Mobile phase B；At 45 minutes, 50% mobile phase A, 50% Mobile phase B；At 52 minutes, 45% flowing
Phase A, 55% Mobile phase B；Mobile phase A and Mobile phase B configuration proportion are prepared by volume.
2. the method for quality control of reinforcing spleen and kidney side according to claim 1, which is characterized in that confession described in step S1
Test sample solution is prepared via a method which to obtain:
S11. take the Radix Astragali of crushing, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Poria cocos, Chinese yam, the fleece-flower root, in nine taste of Semen Cuscutae, semen coicis and Radix Salviae Miltiorrhizae
Medicine, after mixing plus water, 20~50min of immersion are decocted 0.5~3 hour, after filtering again after boiling, and filtrate decompression recycling is spraying dry
It is dry, obtain dried powder；
S12. reinforcing spleen and kidney side's dried powder is weighed, is dissolved in acetonitrile solution or methanol aqueous solution, ultrasonic dissolution, constant volume is in capacity
Bottle, filter membrane are spare.
3. the method for quality control of reinforcing spleen and kidney side according to claim 2, which is characterized in that the specific side of step S12
Method are as follows: weigh reinforcing spleen and kidney side dried powder 56mg, be dissolved in 60% acetonitrile solution of 2mL, ultrasonic dissolution, constant volume is in 5mL capacity
Bottle crosses 0.22 μm of filter membrane, spare.
4. the method for quality control of reinforcing spleen and kidney side according to claim 2, which is characterized in that the specific behaviour of step S11
As: take 15~20 parts of 10~18 parts of 15~20 parts of 25~33 parts of Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Poria cocos, the Chinese yam 13~20 of crushing
Part, 15~20 parts of the fleece-flower root, 12~20 parts of Semen Cuscutae, 15~25 parts of semen coicis, 10~20 parts of Radix Salviae Miltiorrhizae add water after mixing, impregnate
30min is decocted 2 hours, after filtering again after boiling, and filter residue decocts once again with method, is merged filtrate twice, is recovered under reduced pressure, spraying
It is dry, obtain dried powder.
5. the method for quality control of reinforcing spleen and kidney side according to claim 1, which is characterized in that column temperature is 25 DEG C；Flow velocity
For 250 μ L/min；Sample volume is 2 μ L；Chromatographic column select Kinetex C18 analytical column, specification be 2.1mm × 100mm, 1.7 μm.
6. the method for quality control of reinforcing spleen and kidney side according to claim 1, which is characterized in that the liquid described in step S2
Mass spectrograph is also connected with after chromatography, for detecting the molecular weight at shared peak；The mass spectrograph uses electric spray ion source
(ESI), sheath gas is 45 units, and auxiliary gas is 6 units, and spray voltage is -2.5~-3.5kV, and capillary temperature is 300~375
DEG C, sample first uses high-resolution to carry out full scan, second order ms using the scanning of dynamic data dependence, collision energy is set as 20~
45%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610917155.0A CN107064320B (en) | 2016-10-21 | 2016-10-21 | A kind of method of quality control and its finger-print of reinforcing spleen and kidney side |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610917155.0A CN107064320B (en) | 2016-10-21 | 2016-10-21 | A kind of method of quality control and its finger-print of reinforcing spleen and kidney side |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107064320A CN107064320A (en) | 2017-08-18 |
| CN107064320B true CN107064320B (en) | 2019-09-24 |
Family
ID=59616858
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610917155.0A Expired - Fee Related CN107064320B (en) | 2016-10-21 | 2016-10-21 | A kind of method of quality control and its finger-print of reinforcing spleen and kidney side |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107064320B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107907619A (en) * | 2017-10-25 | 2018-04-13 | 中山市中智药业集团有限公司 | A kind of blood concentration quantitative analysis method of radix astragali broken wall medicine materical crude slice active ingredient |
| CN108918696B (en) * | 2018-05-14 | 2021-07-27 | 广东省中医院 | Establishment method and quality control method of spleen-tonifying and lung-benefiting prescription fingerprint |
| CN108918697B (en) * | 2018-05-14 | 2021-07-23 | 广东省中医院 | Method for measuring content of calycosin and calycosin glycoside in spleen-invigorating and lung-benefiting prescription |
| CN112014480B (en) * | 2019-05-28 | 2023-03-28 | 黄河科技学院 | Method for detecting content of effective components in Jiangzhining granules by UPLC-MS/MS |
| CN110455973B (en) * | 2019-08-21 | 2021-08-17 | 劲牌生物医药有限公司 | Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang |
| CN111175428B (en) * | 2020-01-07 | 2022-09-09 | 江苏省中医院 | Method for detecting fingerprint of kidney-tonifying pregnancy-assisting granules |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102579894A (en) * | 2012-02-10 | 2012-07-18 | 广东省中医院 | Chinese materia medica preparation for curing chronic kidney disease and preparation method of Chinese materia medica preparation |
| CN102818861A (en) * | 2012-05-10 | 2012-12-12 | 丽珠医药集团股份有限公司 | Quality control method and application of Qingdu Anshen capsule |
| CN103257191A (en) * | 2013-04-27 | 2013-08-21 | 太极集团重庆涪陵制药厂有限公司 | Method for assaying kidney tonifying and life lengthening capsule fingerprint |
| CN105486790A (en) * | 2015-11-18 | 2016-04-13 | 山西德元堂药业有限公司 | Fingerprint detection method for Shenshuaining granule |
| CN105675750A (en) * | 2016-01-18 | 2016-06-15 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC characteristic chromatogram of Chinese patent medicine 'Yishen Bugu liquid' |
-
2016
- 2016-10-21 CN CN201610917155.0A patent/CN107064320B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102579894A (en) * | 2012-02-10 | 2012-07-18 | 广东省中医院 | Chinese materia medica preparation for curing chronic kidney disease and preparation method of Chinese materia medica preparation |
| CN102818861A (en) * | 2012-05-10 | 2012-12-12 | 丽珠医药集团股份有限公司 | Quality control method and application of Qingdu Anshen capsule |
| CN103257191A (en) * | 2013-04-27 | 2013-08-21 | 太极集团重庆涪陵制药厂有限公司 | Method for assaying kidney tonifying and life lengthening capsule fingerprint |
| CN105486790A (en) * | 2015-11-18 | 2016-04-13 | 山西德元堂药业有限公司 | Fingerprint detection method for Shenshuaining granule |
| CN105675750A (en) * | 2016-01-18 | 2016-06-15 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC characteristic chromatogram of Chinese patent medicine 'Yishen Bugu liquid' |
Non-Patent Citations (3)
| Title |
|---|
| 济生肾气丸的高效液相色谱法特定指纹图谱的研究;胡丹 等;《国际药学研究杂志》;20150430;第42卷(第2期);225-228 * |
| 肾通颗粒HPLC指纹图谱的研究;姜琳琳 等;《中成药》;20070531;第29卷(第5期);625-628 * |
| 补肾方分煎和合煎提取物的指纹图谱比较;史万忠 等;《中成药》;20040131;第26卷(第1期);7-9 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107064320A (en) | 2017-08-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107064320B (en) | A kind of method of quality control and its finger-print of reinforcing spleen and kidney side | |
| CN102539553B (en) | Method for establishing fingerprint spectrum of liver-enhancing medicine | |
| CN105806975A (en) | Method for establishing lonicerae and forsythiae powder UPLC fingerprint spectrum | |
| CN111175428B (en) | Method for detecting fingerprint of kidney-tonifying pregnancy-assisting granules | |
| CN111999423B (en) | Establishment method and application of neck pain granule fingerprint spectrum | |
| CN112903882B (en) | HPLC (high Performance liquid chromatography) characteristic spectrum of Baoyuan decoction preparation and construction method thereof | |
| CN106483213A (en) | The method for building up of Taohe Chengqi decoction composition finger-print and finger-print | |
| CN110596271A (en) | UPLC characteristic spectrum construction method and identification method of radix pseudostellariae medicinal material and radix pseudostellariae standard decoction | |
| CN104316613B (en) | A kind of method for building up of dispelling wind detoxicating capsule finger printing | |
| CN113376273B (en) | Detection method of HPLC (high performance liquid chromatography) characteristic spectrum of Qingxin lotus seed drink and application of characteristic spectrum | |
| CN102539599B (en) | Method for detecting liver-enhancing medicine | |
| CN108459128A (en) | A kind of method of quality control of danggui sini decoction composition | |
| CN108226325A (en) | Roripa montana gives birth to the method for building up of arteries and veins oral liquid composition finger-print | |
| CN107576739A (en) | A kind of HPLC fingerprint atlas detection methods of LONGMU ZHUANGGU KELI | |
| CN109115904B (en) | Construction method and application of UPLC fingerprint of Dingkundan | |
| CN106526000B (en) | The method for building up and finger-print of Shengma Gegen Tang composition finger-print | |
| CN102068553A (en) | Method for constructing high performance liquid chromatographic (HPLC) fingerprint of Rupixiao preparation | |
| CN102590423B (en) | Fingerprint spectrum determination method of ligusticum wallichii tea-blending granular preparation | |
| CN105974030B (en) | The method for building up of triterpenes components finger-print in a kind of GUIZHI FULING JIAONANG | |
| CN101716308A (en) | Quality control method of calculus bovis antihypertensive pills | |
| CN105169234B (en) | A kind of quality determining method of Chinese materia medica preparation that treating diabetic retinopathy | |
| CN108459091A (en) | The method for building up and finger-print of danggui sini decoction finger-print | |
| CN105929096A (en) | Establishment method for HPLC (high performance liquid chromatography) fingerprint of cold treatment medicament | |
| CN102841154B (en) | Quality testing method of Ganshao Xiaoke tablets | |
| CN107894466B (en) | Determination method of HPLC fingerprint of Jinsangqi antitoxic preparation and quality control method of Jinsangqi antitoxic preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190924 Termination date: 20201021 |