CN106442759A - Detection method of polysaccharide content in cassia twig and poria cocos capsules - Google Patents
Detection method of polysaccharide content in cassia twig and poria cocos capsules Download PDFInfo
- Publication number
- CN106442759A CN106442759A CN201610784572.2A CN201610784572A CN106442759A CN 106442759 A CN106442759 A CN 106442759A CN 201610784572 A CN201610784572 A CN 201610784572A CN 106442759 A CN106442759 A CN 106442759A
- Authority
- CN
- China
- Prior art keywords
- content
- detection method
- detection
- anhydrous glucose
- sample fluid
- 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.)
- Pending
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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention belongs to the field of detection of traditional Chinese medicines and in particular relates to a detection method of polysaccharide content in cassia twig and poria cocos capsules. The method comprises the following steps: a) mixing water and contents of the cassia twig and poria cocos capsules; then carrying out ultrasonic treatment and solid-liquid separation in sequence to obtain sediment; b) mixing the sediment with sulfuric acid; carrying out a refluxing reaction to obtain a reaction solution; c) adjusting the pH (Potential of Hydrogen) value of the reaction solution to be 4 to 8, and mixing the reaction solution with ethanol to obtain a sample solution to be detected; d) detecting the sample solution to be detected by adopting a high performance liquid chromatography-evaporative light scattering detector, and calculating according to a detection result and a pre-established standard curve to obtain the content of D-anhydrous dextrose in the sample solution to be detected. The method provided by the invention can be used for evaluating specificity, precision, stability, repeatability and recovery rate, and can be used for evaluating the polysaccharide content in the cassia twig and poria cocos capsules.
Description
Technical field
The invention belongs in field of traditional Chinese medicine detection, more particularly to a kind of GUIZHI FULING JIAONANG polyoses content detection method.
Background technology
By Ramulus Cinnamomi, Poria, Cortex Moutan, Radix Paeoniae Rubra and Semen Persicae, Chinese medicine of the five flavours constitutes GUIZHI FULING JIAONANG altogether, with promoting blood circulation, changes
The stasis of blood, the effect of disappear, cure mainly block, amenorrhea, dysmenorrhea, prolonged lochia caused by married woman's obstruction of collaterals by blood stasis；Hysteromyoma, chronic basin
Chamber inflammation enclosed mass, dysmenorrhea, endometriosis, ovarian cyst is shown in above-mentioned patient；Can also be used for female mammary gland perfume cystic hyperplasia
Disease category obstruction of collaterals by blood stasis disease, symptoms include mastalgia, lump in breast, costa sternaless feeling of distension and oppression；Or belong to stasis blocking bladder card, disease for prostatic hyperplasia
See that urine is not well, lower abdominal distention pain.
Polyoses content in GUIZHI FULING JIAONANG has highly important to the quality for controlling and evaluating GUIZHI FULING JIAONANG
Meaning.Development process being adopted the detection of polyoses content at present, this method specificity is poor, and accuracy is not high more.Therefore, in order to more
Reflect well the inherent quality of GUIZHI FULING JIAONANG, the detection method for studying polyoses content in a kind of new GUIZHI FULING JIAONANG is
Very necessary.
Content of the invention
In view of this, it is an object of the invention to provide in a kind of GUIZHI FULING JIAONANG polyoses content detection method, this
The method specificity that invention is provided is good, accuracy height.
The invention provides in a kind of GUIZHI FULING JIAONANG polyoses content detection detection method, comprise the following steps：
A) supersound process and solid-liquid separation are carried out successively after, mixing water with the content of GUIZHI FULING JIAONANG, is sunk
Starch；
B), the precipitate is mixed with sulphuric acid, back flow reaction, obtains reactant liquor；
C), the pH value of the reactant liquor is adjusted and mixes with ethanol to after 4～8, obtain analyte sample fluid；
D), using HPLC ELSD detector, the analyte sample fluid is detected, according to detection
As a result calculate with the standard curve for pre-building, obtain the content of D- anhydrous glucose in the analyte sample fluid.
Preferably, the content is (0.1～0.5) g with the amount ratio of water：(20～50) mL.
Preferably, the concentration of the sulphuric acid is 2～5mol/mL；
Preferably, the content is (0.1～0.5) g with the amount ratio of sulphuric acid：(10～30) mL.
Preferably, the time of the back flow reaction is 2～6h.
Preferably, in step c), the pH value of the reactant liquor is adjusted to 4.5～6.
Preferably, in step d), the mobile phase for adopting in detection process is triethylamine solution and acetonitrile.
Preferably, in step d), the flow rate of mobile phase in detection process is 0.8～1.2mL/min.
Preferably, in step d), the chromatographic column column temperature in detection process is 40～50 DEG C.
Preferably, in step d), the theoretical cam curve in detection process is calculated by D- anhydrous glucose peak and is not less than 2000.
Compared with prior art, the invention provides in a kind of GUIZHI FULING JIAONANG polyoses content detection method, the party
Method is comprised the following steps：A) after, mixing water with the content of GUIZHI FULING JIAONANG successively supersound process and solid-liquid separation are carried out,
It is precipitated thing；B), the precipitate is mixed with sulphuric acid, back flow reaction, obtains reactant liquor；C), by the pH of the reactant liquor
Value is adjusted and is mixed with ethanol to after 4～8, obtains analyte sample fluid；D), HPLC ELSD detector is adopted
The analyte sample fluid is detected, is calculated according to testing result and the standard curve that pre-builds, obtain described treating test sample
The content of D- anhydrous glucose in product liquid.The present invention is dissolved in GUIZHI FULING JIAONANG with water under ultrasound condition as solvent first
Tolerant middle monosaccharide, disaccharide and beta-schardinger dextrin-；Then solid-liquid separation, obtains the precipitate containing polysaccharide component；Afterwards this is precipitated
Thing and sulphuric acid hybrid reaction, it is D- anhydrous glucose to make the polysaccharide hydrolysis in precipitate；The reactant liquor system for finally reaction being obtained
Become analyte sample fluid, anhydrous to the D- in analyte sample fluid using HPLC (high performance liquid chromatography)-ELSD (evaporat light scattering) method
The content of glucose is detected.D- anhydrous grape as the carbohydrate content in precipitate only has polysaccharide, in analyte sample fluid
Sugar is formed by polysaccharide hydrolysis, and therefore the big I by determining D- anhydrous grape sugared content in analyte sample fluid directly reflects
The height of polyoses content in GUIZHI FULING JIAONANG, so as to realize the evaluation to polyoses content in GUIZHI FULING JIAONANG.To the present invention
The specificity, precision, stability, repeatability of the method for offer and the response rate are evaluated, and as a result show what the present invention was provided
Method specificity is good, precision RSD≤1.16%, stability RSD≤2.21%, and repeated RSD is 1.89%, average recovery
It is 1.39% that rate is 100.11%, response rate RSD, can be used to evaluate the polyoses content in GUIZHI FULING JIAONANG.
Description of the drawings
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
Accompanying drawing to be used needed for technology description is had to be briefly described, it should be apparent that, drawings in the following description are only this
Inventive embodiment, for those of ordinary skill in the art, on the premise of not paying creative work, can also basis
The accompanying drawing of offer obtains other accompanying drawings.
Fig. 1 is the concentration-peak area standard curve of the D- anhydrous glucose that the embodiment of the present invention 1 is provided；
Fig. 2 is the D- anhydrous glucose assay specificity test collection of illustrative plates that the embodiment of the present invention 5 is provided；
Fig. 3 is the chromatogram of the different in flow rate that the embodiment of the present invention 6 is provided and column temperature；
Fig. 4 is the chromatogram of different flow rate of carrier gass, drift tube temperature and chromatographic column that the embodiment of the present invention 6 is provided.
Specific embodiment
Below the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment
The only a part of embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, the common skill in this area
The every other embodiment obtained under the premise of creative work is not made by art personnel, belongs to the model of present invention protection
Enclose.
The invention provides in a kind of GUIZHI FULING JIAONANG polyoses content detection method, comprise the following steps：
A) supersound process and centrifugation are carried out successively after, mixing water with the content of GUIZHI FULING JIAONANG, is precipitated thing；
B), the precipitate is mixed with sulphuric acid, back flow reaction, obtains reactant liquor；
C), the pH value of the reactant liquor is adjusted and mixes with ethanol to after 4～8, obtain analyte sample fluid；
D), using HPLC ELSD detector, the analyte sample fluid is detected, according to detection
As a result calculate with the standard curve for pre-building, obtain the polyoses content in GUIZHI FULING JIAONANG.
In the method that the present invention is provided, remove the monosaccharide in GUIZHI FULING JIAONANG content, disaccharide and β-ring first and paste
Essence, using by the way of be：A) supersound process is carried out successively after, mixing water with the content of GUIZHI FULING JIAONANG and solid-liquid divides
From being precipitated thing.Wherein, the amount ratio of the content and water is preferably (0.1～0.5) g：(20～50) mL, more preferably
For 0.25g：(20～50) mL, most preferably 0.25g：30mL.The frequency of the supersound process is preferably 20～60KHz, more excellent
Elect 30～40KHz as；The power of the supersound process is preferably 200～300W, more preferably 250～260W；At the ultrasound
The time of reason is preferably 10～60min；More preferably 30～40min.The mode of the solid-liquid separation is preferably and is centrifuged；Described from
The rotating speed of the heart preferably 5000～20000r/min, more preferably 10000～15000r/min, most preferably 11000～
13000r/min；The time of the centrifugation is preferably 10～30min, more preferably 15～20min.In the present invention, in order to carry
The degree of purity (reducing monosaccharide, disaccharide and beta-schardinger dextrin-content) of polysaccharide, the precipitate weight that preferred pair is obtained in high precipitate
The operation of step a) carried out again, and the number of times of the repetition is preferably 1～3 time, more preferably 2 times.
Obtain, in the precipitate, the precipitate being mixed with sulphuric acid, carry out back flow reaction.Wherein, the sulphuric acid
Concentration is preferably 2～5mol/mL, more preferably 3mol/mL.The content is preferably (0.1～0.5) with the amount ratio of sulphuric acid
g：(10～30) mL, more preferably 0.25g：(10～30) mL, most preferably 0.25g：20mL.The time of the back flow reaction is excellent
Elect 2～6h, more preferably 3～4h as.During back flow reaction, in GUIZHI FULING JIAONANG, polysaccharide hydrolysis are D- anhydrous glucose, to return
After stream reaction terminates, reactant liquor is obtained.
After obtaining the reactant liquor, the pH value of the reactant liquor is adjusted to 4～8, is preferably adjusted to 4.5～6.At this
In one embodiment of bright offer, using the pH value of reactant liquor described in sodium hydroxide solution condition, the sodium hydroxide solution
Concentration is preferably 35～40wt%.After pH regulator is finished, which is mixed with ethanol, obtain analyte sample fluid.In the present invention, excellent
Choosing first carries out concentrating under reduced pressure to the reactant liquor for completing pH regulator, mixes which with ethanol again afterwards, obtains analyte sample fluid.At this
In invention, preferably first the reactant liquor of concentrating under reduced pressure is mixed with part ethanol, afterwards solid-liquid separation, liquid phase ethanol again with surplus
Mixing, obtains analyte sample fluid.In one embodiment that the present invention is provided, the concentration of alcohol in the analyte sample fluid is 70
～80vol%.In one embodiment that the present invention is provided, the content is excellent with the mass volume ratio of the analyte sample fluid
Elect (0.1～0.5) g as：(50～200) mL, more preferably 0.25g：(50～200) mL, most preferably：0.25g：100mL.
After obtaining the analyte sample fluid, using HPLC ELSD detector to the testing sample
Liquid is detected.In detection process, the mobile phase of employing is preferably triethylamine solution and acetonitrile；The concentration of the triethylamine solution
Preferably 0.1～0.2vol%；The volume ratio of the triethylamine solution and acetonitrile is preferably (10～30)：(90～70), more excellent
Elect 20 as：80.In detection process, flow rate of mobile phase is preferably 0.8～1.2mL/min, more preferably 1mL/min.Detection process
In, chromatographic column is preferably XBridge Amide type chromatographic column；The filler of the chromatographic column is preferably with octadecylsilane key
Close silica gel；The column temperature of chromatographic column is preferably 40～50 DEG C, more preferably 45 DEG C.In detection process, the temperature of drift tube is preferably
90～100 DEG C, more preferably 95 DEG C.In detection process, the gas flow rate of carrier gas is preferably 2～3L/min.In detection process, reason
D- anhydrous glucose peak is pressed by the number of plates and calculate preferably not less than 2000.After detection terminates, D- in analyte sample fluid is obtained anhydrous
The peak area of glucose, according to the peak area of D- anhydrous glucose in the analyte sample fluid and the D- anhydrous grape for pre-building
Concentration-peak area the standard curve of sugar, obtains the content of D- anhydrous glucose in analyte sample fluid.
The present invention sets up mode and is not particularly limited to the concentration-peak area standard curve of the D- anhydrous glucose, can
Set up in such a way：
First, a series of standard solution of D- anhydrous glucose is provided；Then, dissipated using high performance liquid chromatography-evaporative light
Penetrate detector a series of standard solution of D- anhydrous glucose is detected, obtain D- each concentration of anhydrous glucose
The chromatogram of standard solution；Finally, according to the concentration of the corresponding standard solution of the chromatogram, D- anhydrous glucose is obtained
Concentration-peak area standard curve.
In the method for the concentration-peak area standard curve of the above-mentioned acquisition D- anhydrous glucose that the present invention is provided, first
A series of standard solution of the D- anhydrous glucose of variable concentrations is provided, the standard solution is joined by D- anhydrous glucose and ethanol
It is obtained, the concentration of the ethanol is preferably 70～80vol%.In one embodiment that the present invention is provided, a series of differences
The concentration of the standard solution of the D- anhydrous glucose of concentration is 200～2100 μ g/mL；Another in present invention offer is real
Apply in example, a series of concentration of the standard solution of the D- anhydrous glucose of variable concentrations be respectively 201.8 μ g/mL, 504.5
μ g/mL, 807.2 μ g/mL, 1109.9 μ g/mL, 1412.6 μ g/mL, 1715.3 μ g/mL and 2018 μ g/mL.Obtain a series of D-
After the standard solution of anhydrous glucose concentration, using HPLC ELSD detector to a series of D- no
The standard solution of water concentration of glucose is detected, obtains the chromatogram of the D- anhydrous glucose standard solution of each concentration.?
In the present invention, the chromatographic condition when detection chromatographic condition of the standard solution is detected with the analyte sample fluid is consistent,
Will not be described here.After obtaining the chromatogram of D- anhydrous glucose standard solution of each concentration, the present invention is calculated each
The peak area of the D- anhydrous glucose standard solution of concentration, according to the corresponding D- anhydrous grape Standard for Sugars of the peak area that obtains
The concentration of solution, obtains D- anhydrous glucose concentration-peak area standard curve.
As the carbohydrate content in precipitate only has polysaccharide, the D- anhydrous glucose in analyte sample fluid is by polysaccharide hydrolysis
Form, therefore obtain directly reflecting Ramulus Cinnamomi Fu according to the size of its content after D- anhydrous grape sugared content in analyte sample fluid
The height of polyoses content in Siberian cocklebur capsule, so as to realize the evaluation to polyoses content in GUIZHI FULING JIAONANG.
The present invention dissolved under ultrasound condition with water as solvent first monosaccharide in the content of GUIZHI FULING JIAONANG, disaccharide and
Beta-schardinger dextrin-；Then solid-liquid separation, obtains the precipitate containing polysaccharide component；Afterwards by the precipitate and sulphuric acid hybrid reaction,
It is D- anhydrous glucose to make the polysaccharide hydrolysis in precipitate；The reactant liquor for finally obtaining reaction makes analyte sample fluid, adopts
HPLC (high performance liquid chromatography)-ELSD (evaporative light scattering detection) method is entered to the content of the D- anhydrous glucose in analyte sample fluid
Row detection.As the carbohydrate content in precipitate only has polysaccharide, the D- anhydrous glucose in analyte sample fluid is by polysaccharide hydrolysis
Form, therefore the big I by determining D- anhydrous grape sugared content in analyte sample fluid directly reflects in GUIZHI FULING JIAONANG
The height of polyoses content, so as to realize the evaluation to polyoses content in GUIZHI FULING JIAONANG.Ramulus Cinnamomi Fu is evaluated in this way
In Siberian cocklebur capsule, polyoses content has preferable specificity and higher accuracy.Specificity, essence to the method for present invention offer
Density, stability, repeatability and the response rate are evaluated, and as a result show that the method specificity that the present invention is provided is good, precision
RSD≤1.16%, stability RSD≤2.21%, it is the 100.11%, response rate that repeated RSD is 1.89%, average recovery rate
RSD is 1.39%, can be used to evaluate the polyoses content in GUIZHI FULING JIAONANG.
For the sake of becoming apparent from, it is described in detail below by following examples.
In the present invention, following embodiments are directed to use with following instrument and reagent：
1st, instrument：
Mettler AE240 electronic analytical balance；Sartorius BP211D electronic analytical balance；KQ-250DB numerical control surpasses
Sound cleaning device；HH digital display thermostat water bath；
High performance liquid chromatography-ELSD detector：(1) 1100 high performance liquid chromatograph of Agilent, ELSD detector；(2)
1200 high performance liquid chromatograph of Agilent, ELSD detector.
2nd, reagent：
GUIZHI FULING JIAONANG (lot number：140301、140701、140801、140901；The limited public affairs of Jiangsu Kang Yuan Pharmaceutical share
Department)；D- anhydrous glucose (National Institute for Food and Drugs Control, lot number：1110833-201205, for assay)；Second
Nitrile (chromatographically pure, upper Asterias amurensis Lutken can biochemical company limited)；Ultra-pure water (is made by oneself)；It is pure that other reagents are analysis.
Embodiment 1
Criterion curve
1st, chromatographic condition
Chromatographic column：Xbridge Amide (3.5 μm, 4.6 × 150mm)；With octadecylsilane chemically bonded silica as filling
Agent；With acetonitrile -0.1vol% triethylamine solution (volume ratio=80:20) it is mobile phase；45 DEG C of chromatographic column column temperature；Flow rate of mobile phase
1.0mL/min；ELSD：95 DEG C of drift tube temperature；Gas of carrier gas flow velocity 2.0L/min；Theoretical cam curve is in terms of D- anhydrous glucose
Calculate and should be not less than 2000.
2nd, the concentration-peak area standard curve of D- anhydrous glucose is set up
Take D- anhydrous glucose appropriate, accurately weighed, plus 80vol% ethanol make 201.8 μ g/mL, 504.5 μ g/mL,
807.2 μ g/mL, 1109.9 μ g/mL, 1412.6 μ g/mL, 1715.3 μ g/mL, 2018 μ g/mL D- anhydrous grape Standard for Sugars molten
Liquid, each 10 μ L of the above-mentioned solution of accurate absorption, injects high performance liquid chromatography-ELSD detector, is detected, record chromatographic peak respectively
Peak area.The standard solution of each concentration is detected twice, with sample size (μ g/mL) logarithm as abscissa (X), and that detected twice is flat
The logarithm of all peak areas is vertical coordinate (Y), draws standard curve, as a result as shown in table 1 and Fig. 1.Table 1 is the D- that the present invention is provided
Result investigated by anhydrous glucose linear relationship, and Fig. 1 is the concentration-peak area of the D- anhydrous glucose that the embodiment of the present invention 1 is provided
Standard curve.
Result investigated by table 1D- anhydrous glucose linear relationship
Regression equation is calculated according to table 1 and correlation coefficient is：Y=1.8242X+1.5221；R2=0.9998.It can be seen that, D-
Anhydrous glucose sample size is in good linear relation between 201.8～2018 μ g/mL.The results are shown in Table 1 and Fig. 1.
Embodiment 2
The evaluation of polyoses content in GUIZHI FULING JIAONANG
1st, the preparation of analyte sample fluid：
Take 10 GUIZHI FULING JIAONANG (lot numbers：140301) content, accurately weighed, finely ground, about 0.25g is taken, accurate
Weighed, put in conical flask with cover, add purified water 30mL.Supersound process (40KHz, 250W) 30 minutes, lets cool, centrifugation
(11000r/min, 15min), abandoning supernatant, precipitation adds purified water supersound process 30min again, be centrifuged (11000r/min,
15min), remaining sample in conical flask adds 30mL purified water is multiple on a small quantity to be transferred in centrifuge tube, is centrifuged (11000r/
Min, 15min), centrifuged deposit adds 3mol/mL sulphuric acid 20mL to be repeatedly transferred in Backflow bottle on a small quantity, flows back in boiling water bath
3h, lets cool, plus 35wt% sodium hydroxide adjusts pH value to 4.5～6.0, concentrating under reduced pressure, plus ethanol dissolving, makes solution ethanol concentration
For 70～80vol%, let cool, centrifugation, solution adds 80vol% ethanol to be settled to 100mL, shakes up, obtain GUIZHI FULING JIAONANG
Analyte sample fluid.
2nd, the measure of D- anhydrous grape sugared content
6 parts of the analyte sample fluid for preparing according to the method described above is taken, respectively each 5 μ L of the above-mentioned analyte sample fluid of accurate absorption, note
Enter high performance liquid chromatography-ELSD detector, detected according to the chromatographic condition in embodiment 1, record chromatographic peak peak area, root
The concentration of the D- anhydrous glucose that sets up with embodiment 1 according to the peak area-peak area standard curve, is calculated testing sample
The content of D- anhydrous glucose in liquid, as a result as shown in table 2：
The cubage result of table 2D- anhydrous glucose
| Numbering | 1 (wt%) | 2 (wt%) | 3 (wt%) | 4 (wt%) | 5 (wt%) | 6 (wt%) | Average content | RSD (%) |
| D- anhydrous glucose | 35.19 | 35.93 | 35.33 | 36.69 | 36.81 | 36.33 | 36.05 | 1.89 |
It can be seen from Table 2 that, the method repeatability is good, and in analyte sample fluid, the average content of D- anhydrous glucose is
36.05wt%, RSD are 1.89%.
Embodiment 3
System suitability
1st, the preparation of reference substance solution
Take D- anhydrous glucose reference substance appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous grape containing D-
The solution of sugared 0.8mg, obtains final product reference substance solution.
2nd, the preparation of need testing solution：
Preparation method according to analyte sample fluid described in embodiment 2 is obtained need testing solution.
3rd, determine
Accurate absorption 5 μ L of reference substance solution, injects high performance liquid chromatography-ELSD detector, according to the chromatograph in embodiment 1
Condition is detected, continuously repeats sample introduction 5 times, calculates the relative standard deviation of peak area measurement value；Accurate absorption test sample is molten
5 μ L of liquid, injects high performance liquid chromatography-ELSD detector, is detected according to the chromatographic condition in embodiment 1, calculates to be measured group
Theoretical cam curve, separating degree and the tailing factor for dividing.As a result as shown in table 3：
3 GUIZHI FULING JIAONANG system suitability major parameter result table of table
Under these conditions, tailing factor is the chromatographic system peak area repeatability of 0.92～0.95, D- anhydrous glucose
Relative standard deviation is 0.53%, calculates theoretical plate number with D- anhydrous glucose peak and is more than 2000, for guaranteeing the standard of quantitative analyses
Really, it is stipulated that number of theoretical plate is pressed D- anhydrous glucose peak and must not calculate less than 2000.
Embodiment 4
The impact of analyte sample fluid preparation process conditional parameter
1st, the impact of water extraction number of times
Take 10 GUIZHI FULING JIAONANG (lot numbers：140301) content, accurately weighed, finely ground, about 0.25g is taken, accurate
Weighed, put in conical flask with cover, add purified water 30mL.Supersound process (40KHz, 250W) 30 minutes, lets cool, centrifugation
(11000r/min, 15min), obtains supernatant (i.e. one time extracting solution) and precipitation, and precipitation adds purified water supersound process again
30min, is centrifuged (11000r/min, 15min), obtains supernatant (i.e. secondary raffinate) and precipitation, and precipitation in conical flask is added
Enter that 30mL purified water is multiple on a small quantity to be transferred in centrifuge tube, be centrifuged (11000r/min, 15min), obtain supernatant (i.e. three times
Extracting solution) and precipitation.To the Fructose in extracting solution, secondary raffinate and three extracting solution, glucose, sucrose and β-ring paste
Smart content is measured, as a result as shown in table 4：
4 extraction time of table investigates result
| Extraction time | Fructose (wt%) | Glucose (wt%) | Sucrose (wt%) | Beta-schardinger dextrin-(wt%) |
| 1st time | 6.01 | 7.43 | 16.43 | 7.68 |
| 2nd time | 0.57 | 0.87 | 1.06 | 3.16 |
| 3rd time | No | No | No | 0.14 |
Experimental result shows：With after water extraction three times substantially will monosaccharide, disaccharide and cyclodextrin remove clean.
2nd, the impact of sulfuric acid concentration
The GUIZHI FULING JIAONANG content that lot number is 140301 is taken, according to the preparation method of analyte sample fluid in embodiment 2,
1. and 2. select the sulphuric acid of 2mol/mL, 3mol/mL, 4mol/mL and 5mol/mL to prepare 2 parts of analyte sample fluids respectively, be labeled as.
The each 5 μ L of the above-mentioned analyte sample fluid of accurate absorption, injects high performance liquid chromatography-ELSD detector, according to the color in embodiment 1 respectively
Spectral condition is detected, record chromatographic peak peak area, the D- anhydrous glucose that is set up with embodiment 1 according to the peak area dense
Degree-peak area standard curve, is calculated the content of D- anhydrous glucose in analyte sample fluid, as a result as shown in table 5：
Impact of the 5 variable concentrations sulphuric acid of table to D- anhydrous glucose assay
It can be seen from Table 5 that, different sulfuric acid concentrations have certain impact to polysaccharide hydrolysis, and 3mol/mL is optium concentration.
3rd, the impact of different pH value
The GUIZHI FULING JIAONANG content that lot number is 140301 is taken, according to the preparation method of analyte sample fluid in embodiment 2,
At " plus 35wt% sodium hydroxide is neutralized to PH for 4.5～6 ", different pH values are adjusted to respectively, by the response rate to investigate not
With impact of the pH value to D- anhydrous glucose content detection；The each 5 μ L of the above-mentioned analyte sample fluid of accurate absorption, injects efficient liquid respectively
Phase chromatograph-ELSD detector, is detected according to the chromatographic condition in embodiment 1, records chromatographic peak peak area, according to the peak
The concentration of the D- anhydrous glucose that area is set up with embodiment 1-peak area standard curve, is calculated in analyte sample fluid D- no
The content of water glucose, and then the response rate is calculated, as a result as shown in table 6.In table 6, " sample weighting amount " refers to prepare test sample molten
The amount of the GUIZHI FULING JIAONANG content for being weighed during liquid, " content " refers to that the D- for containing in the content for weighing in theory is anhydrous
The amount of glucose sugar, " addition " refers to the amount of the D- anhydrous glucose reference substance for adding, and " measured value " refers to need testing solution
In the amount of D- anhydrous glucose that measures.
6 difference impact of the pH to the response rate of table
| Numbering | PH value | Sample weighting amount (mg) | Content (mg) | Addition (mg) | Measured value (mg) | The response rate (%) |
| 1 | 10.9 | 125.8 | 48.90 | 38.11 | 76.44 | 72.28 |
| 2 | 4.11 | 125.2 | 48.67 | 38.06 | 80.73 | 84.25 |
| 3 | 4.48 | 125.6 | 48.82 | 38.08 | 81.04 | 84.61 |
| 4 | 4.89 | 124.6 | 48.43 | 38.15 | 80.95 | 85.26 |
| 5 | 6.02 | 125.6 | 48.82 | 38.02 | 80.89 | 84.35 |
| 6 | 5.59 | 126.2 | 49.05 | 38.13 | 80.65 | 82.89 |
Can be seen that, by the data in table 6, the hydrolysis that basic conditions can destroy polysaccharide.
4th, the impact of different return times
The GUIZHI FULING JIAONANG content that lot number is 140301 is taken, according to the preparation method of analyte sample fluid in embodiment 2,
The return time of 2h, 3h, 4h, 5h and 6h is selected to prepare analyte sample fluid respectively.Precision draws above-mentioned analyte sample fluid each 5 respectively
μ L, injects high performance liquid chromatography-ELSD detector, is detected according to the chromatographic condition in embodiment 1, records chromatograph peak-to-peak face
Product, the concentration-peak area standard curve of the D- anhydrous glucose that is set up with embodiment 1 according to the peak area, it is calculated and treats
The content of D- anhydrous glucose in sample liquid is surveyed, as a result as shown in table 7：
The different return times of table 7 investigate result of the test
| Numbering | Return time | Content (wt%) |
| 1 | 2h | 36.53 |
| 2 | 3h | 40.49 |
| 3 | 4h | 39.66 |
| 4 | 5h | 34.96 |
| 5 | 6h | 32.93 |
It can be seen from Table 5 that, different return times have certain impact to polysaccharide hydrolysis, and 3h is optimal return time.
Embodiment 5
Specificity is tested
1st, the preparation of reference substance solution：
Take D- anhydrous glucose reference substance appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous grape containing D-
The solution of sugared 0.8mg, obtains final product reference substance solution.
2nd, the preparation of need testing solution：
Preparation method according to analyte sample fluid described in embodiment 2 is obtained need testing solution.
3rd, the preparation of negative need testing solution
In addition to sample is not added with, feminine gender need testing solution is obtained by need testing solution preparation method.
4th, determine
Accurate absorption 5 μ L of reference substance solution, injects high performance liquid chromatography-ELSD detector, according to the chromatograph in embodiment 1
Condition is detected, as a result as shown in Fig. 2 Fig. 2 is the D- anhydrous glucose assay specificity that the embodiment of the present invention 5 is provided
Test collection of illustrative plates 1 is negative test sample in Fig. 2,2 is reference substance, 3 be.As seen in Figure 2, the present invention is provided
The specificity of method is good.
Embodiment 6
Serviceability test
1st, the impact of different flow rate of mobile phase and chromatographic column column temperature
Take D- anhydrous glucose reference substance appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous grape containing D-
The solution of 1000 μ g of sugar, obtains final product reference substance solution.
Precision draws above-mentioned 5 μ L of reference substance solution, injects high performance liquid chromatography-ELSD detector, according in embodiment 1
Chromatographic condition, respectively under the conditions of the flow rate of mobile phase of 0.9mL/min, 1.0mL/min and 1.1mL/min, and 43 DEG C, 45 DEG C
With detected under the conditions of 47 DEG C of flow rate of mobile phase, record chromatographic peak peak area, as a result as shown in figure 3, Fig. 3 is reality of the present invention
Apply example 6 offer different in flow rate and column temperature chromatogram, in Fig. 31 be flow velocity=1.0mL/min, T=47 DEG C, 2 be flow velocity=
1.0mL/min, T=43 DEG C, 3 is flow velocity=1.1mL/min, T=45 DEG C, and 4 is flow velocity=1.0mL/min, T=45 DEG C, 5 for flowing
Speed=0.9mL/min, T=45 DEG C.As seen in Figure 3 flow rate of mobile phase and chromatographic column column temperature to the appearance time of sample and
Peak type has certain impact, and when wherein flow velocity is 45 DEG C of 1.0mL/min, column temperature, peak type is preferable.
2nd, the impact of different flow rate of carrier gass, drift tube temperature, different instruments and different chromatographic columns
Take D- anhydrous glucose reference substance appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous grape containing D-
The solution of 1000 μ g of sugar, obtains final product reference substance solution.
Precision draws above-mentioned 5 μ L of reference substance solution, injects high performance liquid chromatography-ELSD detector, according in embodiment 1
Chromatographic condition, respectively under the conditions of the flow rate of carrier gas of 2.0L/min, 2.5L/min and 3.0mL/min, and 90 DEG C, 95 DEG C and
Under the conditions of 100 DEG C of drift tube temperature, and detected under the conditions of same model difference numbering chromatographic column, recorded chromatograph peak-to-peak
Area, as a result as shown in figure 4, the different flow rate of carrier gass that Fig. 4 is the embodiment of the present invention 6 to be provided, drift tube temperature and different pillars
Chromatogram, in Fig. 41 be flow rate of carrier gas 2.0L/min, 95 DEG C of drift pipe temperature, 2 be flow rate of carrier gas 2.5L/min, drift Guan Wen
95 DEG C of degree, 3 is flow rate of carrier gas 3.0L/min, 95 DEG C of drift pipe temperature, and 4 is flow rate of carrier gas 2.0L/min, 90 DEG C of drift tube temperature,
5 is flow rate of carrier gas 2.0L/min, 100 DEG C of drift tube temperature, and 6 is flow rate of carrier gas 2.0L/min, 95 DEG C of drift tube temperature, same type
Number different chromatograph pillars.Flow rate of carrier gas, drift tube temperature, different instruments and different chromatographic columns are to sample as seen in Figure 4
Appearance time and peak type impact unobvious.
Embodiment 7
Precision test
1st, the preparation of reference substance solution：
Take D- anhydrous glucose reference substance appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous grape containing D-
The solution of sugared 0.5045mg, 1.1099mg, 1.7153mg, obtains basic, normal, high concentrations control product solution.
The accurate each 10 μ L of above-mentioned basic, normal, high concentrations control product solution, the injection high performance liquid chromatography-ELSD of drawing is examined respectively
Device is surveyed, is detected according to the chromatographic condition in embodiment 1, repeat sample detection 6 times, sample introduction precision is calculated, as a result as table 8
Shown：
The basic, normal, high concentrations control product Precision test result of table 8
It can be seen from Table 8 that, it is 1.16%, 0.32%, 0.30% that RSD is respectively.As a result show that instrument precision is good
Good.
Embodiment 8
Stability test
1st, the preparation of reference substance solution：
Take D- anhydrous glucose appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous glucose containing D- 0.8mg
Solution, obtain final product reference substance solution.
2nd, the preparation of need testing solution：
Preparation method according to analyte sample fluid described in embodiment 2 is obtained need testing solution.
3rd, test
Take same need testing solution (lot number：140301), respectively at 0h, 2h, 3h, 5h, 7h, 9h, 11h, 13h, 15h,
17h, 19h inject high performance liquid chromatography-ELSD detector, are detected according to the chromatographic condition in embodiment 1,5 μ of each sample introduction
L, records chromatographic peak peak area, and calculates RSD value；Separately take same reference substance solution, respectively at 0h, 2h, 4h, 6h, 8h, 10h,
12h, 19h, 21h inject high performance liquid chromatography-ELSD detector, are detected according to the chromatographic condition in embodiment 1, enter every time
5 μ L of sample, records chromatographic peak peak area, and calculates RSD value.The results are shown in Table 9,10：
9 need testing solution stability test result of table
10 reference substance solution stability test result of table
By table 9 and 10 as can be seen that the RSD that the RSD of need testing solution is 2.21%, reference substance solution is 1.61.Say
Bright need testing solution is basicly stable in 19 hours, and reference substance solution is basicly stable in 21h.
Embodiment 9
Recovery test
Take D- anhydrous glucose appropriate, accurately weighed, plus 80vol% ethanol makes every 1mL anhydrous glucose containing D- 0.8mg
Reference substance solution.
Take the sample (lot number of known content：The content of 140301, D- anhydrous glucose is 36.05wt%) about 0.125g,
Accurately weighed, parallel weigh 9 parts, 3 parts one group, put in conical flask with cover, add purified water 30mL.Supersound process (40KHz,
250W) 30 minutes, let cool, be centrifuged (11000r/min, 15min), abandoning supernatant, precipitation adds purified water supersound process again
30min, is centrifuged (11000r/min, 15min), and remaining sample in conical flask adds 30mL purified water is multiple on a small quantity to be transferred to
In centrifuge tube, it is centrifuged (11000r/min, 15min), centrifuged deposit adds 3mol/mL sulphuric acid 20mL to be repeatedly transferred to backflow on a small quantity
In bottle, each group is separately added into D- anhydrous glucose reference substance solution, and flow back in boiling water bath 3h, lets cool, plus 35wt% hydroxide
Sodium neutralization (PH be 4.5-6.0), recovered under reduced pressure is to certain volume, plus ethanol dissolving, make solution ethanol concentration for 70～
80vol%, lets cool, centrifugation, and solution adds 80vol% ethanol to be settled to 100mL, shakes up, obtains final product mark-on and treat test sample.
The above-mentioned mark-on of accurate absorption treats each 5 μ L of test sample respectively, injects high performance liquid chromatography-ELSD detector, according to enforcement
Chromatographic condition in example 1 is detected, records chromatographic peak peak area, anhydrous with D- that embodiment 1 is set up according to the peak area
The concentration of glucose-peak area standard curve, is calculated the content of D- anhydrous glucose, as a result as shown in table 11.In table 11,
" sample weighting amount " refers to prepare the amount of the GUIZHI FULING JIAONANG content for being weighed during need testing solution, and it is interior that " content " refers to weigh
The amount of the D- anhydrous glucose sugar for containing in tolerant in theory, " addition " refers to the D- anhydrous glucose reference substance for adding
Amount, " measured value " refers to the amount of the D- anhydrous glucose for measuring in need testing solution, the response rate=(content+addition)/determine
Value } × 100%.
Table 11D- anhydrous glucose average recovery result of the test
By table 11 as can be seen that it is the side that present invention offer 1.39%, is described for 100.11%, RSD to evaluate the response rate
Method accuracy is higher.
Embodiment 10
The evaluation of polyoses content in GUIZHI FULING JIAONANG
Precision weighs tri- batches of (lot numbers of 0.25g respectively：140701st, 140801,140901) content of GUIZHI FULING JIAONANG,
1. and 2. two parts are weighed per batch sample, is labeled as, being obtained according to the preparation method of analyte sample fluid described in embodiment 2 to be measured
Sample liquid.
The each 5 μ L of the above-mentioned analyte sample fluid of accurate absorption, injects high performance liquid chromatography-ELSD detector, according to enforcement respectively
Chromatographic condition in example 1 is detected, records chromatographic peak peak area, anhydrous with D- that embodiment 1 is set up according to the peak area
The concentration of glucose-peak area standard curve, is calculated the content of D- anhydrous glucose, as a result as shown in table 12：
12 different batches sample size measurement result of table
| Lot number | Content is 1. | Content is 2. | Average content (%) | RSD (%) |
| 140701 | 39.31 | 38.76 | 39.03 | 0.98 |
| 140801 | 40.14 | 38.85 | 39.49 | 2.32 |
| 140901 | 38.62 | 37.27 | 37.95 | 2.52 |
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. in a kind of GUIZHI FULING JIAONANG polyoses content detection detection method, comprise the following steps：
A) supersound process and solid-liquid separation are carried out successively after, mixing water with the content of GUIZHI FULING JIAONANG, is precipitated thing；
B), the precipitate is mixed with sulphuric acid, back flow reaction, obtains reactant liquor；
C), the pH value of the reactant liquor is adjusted and mixes with ethanol to after 4～8, obtain analyte sample fluid；
D), using HPLC ELSD detector, the analyte sample fluid is detected, according to testing result
Calculate with the standard curve for pre-building, obtain the content of D- anhydrous glucose in the analyte sample fluid.
2. detection method according to claim 1, it is characterised in that the amount ratio of the content and water for (0.1～
0.5)g：(20～50) mL.
3. detection method according to claim 1, it is characterised in that the concentration of the sulphuric acid be；
4. detection method according to claim 3, it is characterised in that the amount ratio of the content and sulphuric acid for (0.1～
0.5)g：(10～30) mL.
5. detection method according to claim 1, it is characterised in that the time of the back flow reaction be.
6. detection method according to claim 1, it is characterised in that in step c), the pH value of the reactant liquor is adjusted
To 4.5～6.
7. detection method according to claim 1, it is characterised in that in step d), the mobile phase for adopting in detection process
For triethylamine solution and acetonitrile.
8. detection method according to claim 1, it is characterised in that the flow rate of mobile phase in step d), in detection process
For 0.8～1.2mL/min.
9. detection method according to claim 1, it is characterised in that the chromatographic column column temperature in step d), in detection process
For 40～50 DEG C.
10. the detection method according to any one of claim 1～9, it is characterised in that in step d), in detection process
Theoretical cam curve is calculated by D- anhydrous glucose peak and is not less than 2000.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610784572.2A CN106442759A (en) | 2016-08-29 | 2016-08-29 | Detection method of polysaccharide content in cassia twig and poria cocos capsules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610784572.2A CN106442759A (en) | 2016-08-29 | 2016-08-29 | Detection method of polysaccharide content in cassia twig and poria cocos capsules |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106442759A true CN106442759A (en) | 2017-02-22 |
Family
ID=58091503
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610784572.2A Pending CN106442759A (en) | 2016-08-29 | 2016-08-29 | Detection method of polysaccharide content in cassia twig and poria cocos capsules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106442759A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122728A (en) * | 2019-12-24 | 2020-05-08 | 江苏康缘药业股份有限公司 | Detection method of cassia twig and poria cocos capsules |
| CN114324659A (en) * | 2021-12-29 | 2022-04-12 | 江苏海悦康医药科技有限公司 | Method for detecting organic impurities in gamma cyclodextrin |
| CN115219628A (en) * | 2022-07-19 | 2022-10-21 | 东阿阿胶股份有限公司 | Pretreatment method of compound donkey-hide gelatin slurry and detection method of oligosaccharide in compound donkey-hide gelatin slurry |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101539550A (en) * | 2009-04-28 | 2009-09-23 | 李绍平 | Qualitative and quantitative analysis method for polyoses |
| CN104730009A (en) * | 2015-02-27 | 2015-06-24 | 浙江经贸职业技术学院 | Method for measuring polysaccharide content in tea flower |
| CN105136951A (en) * | 2015-08-07 | 2015-12-09 | 华中农业大学 | Rapid quantitative method for tea polysaccharide monosaccharide composition |
-
2016
- 2016-08-29 CN CN201610784572.2A patent/CN106442759A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101539550A (en) * | 2009-04-28 | 2009-09-23 | 李绍平 | Qualitative and quantitative analysis method for polyoses |
| CN104730009A (en) * | 2015-02-27 | 2015-06-24 | 浙江经贸职业技术学院 | Method for measuring polysaccharide content in tea flower |
| CN105136951A (en) * | 2015-08-07 | 2015-12-09 | 华中农业大学 | Rapid quantitative method for tea polysaccharide monosaccharide composition |
Non-Patent Citations (4)
| Title |
|---|
| QUAN ZHENG ET AL: "Optimization for ultrasound-assisted extraction of polysaccharides with chemical composition and antioxidant activity from the Artemisia sphaerocephala Krasch seeds", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| 木海鸥 等: "高效液相色谱法测定人参渣中人参多糖的含量", 《浙江中西医结合杂志》 * |
| 李家春 等: "过程控制技术对桂枝茯苓胶囊质量的影响", 《中国中药杂志》 * |
| 祁俊 等: "UV-分光光度计法和HPLC-ELSD法同时测定健脾止泻胶囊中多糖含量", 《中国医药导报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111122728A (en) * | 2019-12-24 | 2020-05-08 | 江苏康缘药业股份有限公司 | Detection method of cassia twig and poria cocos capsules |
| CN111122728B (en) * | 2019-12-24 | 2021-03-26 | 江苏康缘药业股份有限公司 | Detection method of cassia twig and poria cocos capsules |
| CN114324659A (en) * | 2021-12-29 | 2022-04-12 | 江苏海悦康医药科技有限公司 | Method for detecting organic impurities in gamma cyclodextrin |
| CN115219628A (en) * | 2022-07-19 | 2022-10-21 | 东阿阿胶股份有限公司 | Pretreatment method of compound donkey-hide gelatin slurry and detection method of oligosaccharide in compound donkey-hide gelatin slurry |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103728387B (en) | Ginseng-astragalus blood-sugar lowering preparation HPLC standard finger-print and construction method thereof | |
| CN106442759A (en) | Detection method of polysaccharide content in cassia twig and poria cocos capsules | |
| CN110632208B (en) | Detection method for main components of traditional Chinese medicine composition for clearing lung, eliminating phlegm, relieving cough and asthma | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN102445500B (en) | Method for detecting macromolecular substances in Yiqifumai injection | |
| CN107402265B (en) | Detection method of Kangyun granule fingerprint | |
| CN106324161A (en) | Quality detection method for traditional Chinese medicine composition capable of treating diabetic nephropathy | |
| CN104730158B (en) | A kind of content assaying method of JIAWEI HUOXIANG ZHENGQI RUANJIAONANG | |
| CN107991399A (en) | Method that is a kind of while measuring 31 kinds of components in compound Salviae Miltiorrhizae extract or related medicinal material | |
| CN103808835A (en) | Method for simultaneously measuring contents of 10 chemical components in four-substance soup decoction by HPLC-DAD (high performance liquid chromatography-diode array detection) method | |
| CN109932441B (en) | Method for establishing HPLC fingerprint of Shuganning injection | |
| CN106053702B (en) | A kind of multicomponent content assaying method of JIAWEI XIAOYAOWAN | |
| CN109633043A (en) | A kind of method for building up and its standard diagram of gadol injection HPLC finger-print | |
| CN106770876A (en) | The characteristic spectrum of Radix Astragali Jianwei is set up and detection method | |
| CN106802327A (en) | It is a kind of to set up the method that youngster rushes down the finger-print for stopping pharmaceutical preparation | |
| CN110274978A (en) | The more saponin constituent content assaying methods of Radix Notoginseng in a kind of QISHEN YIQI DIWAN | |
| CN109521106A (en) | Method that is a kind of while detecting four kinds of component contents in five composition granule of Radix Astragali ramulus cinnamomi | |
| CN105510452B (en) | Multi-target ingredient assay, fingerprint map construction and the preparation method of liver-benefiting eye-brightening oral liquid | |
| CN110568099B (en) | Fingerprint spectrum construction method of radix acanthopanacis senticosi, radix angelicae sinensis and radix astragali refining agent and multi-index component synchronous content determination method | |
| CN102435693B (en) | Method for controlling quality of midbody during preparation process of Shenmai injection | |
| CN106124682A (en) | A kind of composition method of inspection of Radix Et Caulis Acanthopanacis Senticosi injection | |
| CN108459129A (en) | A kind of method of quality control of Tetrandra and Poria Decoction composition | |
| CN106290643A (en) | A kind of method of quality control of Chinese medicine Herba Schizonepetae anti-lung cancer activity composition | |
| CN104569217B (en) | The method for building up of gold Rong Xiaopi Granules finger-print | |
| CN105301140B (en) | A kind of injection component detection method promoting blood circulation and removing obstruction in channels |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170222 |
|
| RJ01 | Rejection of invention patent application after publication |