CN107064352A - Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method - Google Patents
Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method Download PDFInfo
- Publication number
- CN107064352A CN107064352A CN201710233582.1A CN201710233582A CN107064352A CN 107064352 A CN107064352 A CN 107064352A CN 201710233582 A CN201710233582 A CN 201710233582A CN 107064352 A CN107064352 A CN 107064352A
- Authority
- CN
- China
- Prior art keywords
- rsd
- blood fat
- similarity
- rutin
- chrysin
- 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
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)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method, it is characterised in that it comprises the following steps：(1) preparation of reference substance solution；(2) preparation of need testing solution；(3) determination method：Precision is drawn in reference substance solution and each 10 μ l of need testing solution, injection high performance liquid chromatograph, is measured according to setting chromatographic condition, is recorded HPLC chromatogram；(4) precision test：Take with a collection of need testing solution, continuous sample introduction 6 times, calculate the relative retention time of 1~No. 30 shared fingerprint peaks and the RSD values of relative peak area, while calculating the similarity of each chromatographic fingerprinting with similarity evaluation software.As a result RSD values are respectively less than 3%, and similarity is all higher than 0.990.The inventive method is simple to operation, and quality that can be quickly and accurately to Beijing enamel ware blood fat regulating capsule is measured, precision, reproducible, and the degree of accuracy is high, and the rate of recovery is good.
Description
Technical field
The present invention relates to Chinese medicine fields of measurement, more particularly to a kind of Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method.
Background technology
Beijing enamel ware blood fat regulating capsule is mainly made up of Exocarpium Citri Grandis, ginkgo leaf, gynostemma pentaphylla and propolis, with replenishing qi to invigorate the spleen, promoting blood circulationization
Effect of the stasis of blood, eliminating the phlegm dampness removing, is mainly used in treating qi-asthenia phlegm stasis blocking numbness type hyperlipidemia.Huang is mainly contained in Beijing enamel ware blood fat regulating capsule
The compositions such as ketone, saponins, organic acid, amino acids and polysaccharide, this research adjusts fat using high performance liquid chromatography to Beijing enamel ware
Capsule carries out finger-print research, and Exocarpium Citri Grandis, ginkgo leaf, gynostemma pentaphylla and propolis medicinal material and its composition are belonged to, and sets up
Beijing enamel ware blood fat regulating capsule efficient liquid-phase chromatograph finger print atlas；And carry out multi-target ingredient on the basis of Beijing enamel ware blood fat regulating capsule finger-print
Quantitative analysis, carries out the research of Beijing enamel ware blood fat regulating capsule multi-target ingredient quantitative finger print atlas, comprehensively to reflect the inherent matter of preparation
Amount.
The content of the invention
It is an object of the present invention in view of the above-mentioned problems, provide a kind of Beijing enamel ware blood fat regulating capsule finger-print quality control side
Method.
The technical scheme that is used to achieve the above object of the present invention for：
A kind of Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method, it comprises the following steps：
(1) preparation of reference substance solution：Take aurantiin reference substance, control substance of Rutin, Chrysin reference substance and Galangin
Appropriate reference substance, it is accurately weighed, plus methanol be made every 1mL containing the μ g of aurantiin 80.23, the μ g of rutin 68.12, the μ g of Chrysin 22.82,
The μ g of Galangin 45.57 mixed solution, is produced；
(2) preparation of need testing solution：This product content about 0.25g is taken, is put in conical flask with cover, precision adds methanol
25mL, weighed weight ultrasonically treated 30 minutes, is let cool, then weighed weight, and the weight of less loss is supplied with methanol, is shaken up, filtration,
Subsequent filtrate is taken, is produced；
(3) determination method：Precision is drawn in reference substance solution and each 10 μ l of need testing solution, injection high performance liquid chromatograph,
It is measured according to setting chromatographic condition, records HPLC chromatogram；
(4) precision test：Take with a collection of need testing solution, continuous sample introduction 6 times, calculate 1~No. 30 shared fingerprint peaks
Relative retention time and relative peak area RSD values, while calculating the phase of each chromatographic fingerprinting with similarity evaluation software
Like degree.As a result RSD values are respectively less than 3%, and similarity is all higher than 0.990.
It is described set chromatographic condition as：
Chromatographic column：SHISEIDO Capcell pak MG C18 posts (4.6 × 250mm, 5 μm)；Mobile phase：Using methanol as
Mobile phase A, using 0.1% phosphate aqueous solution as Mobile phase B, gradient elution is carried out by table 2；Flow velocity：1.0mL/min；Column temperature：25
℃；Detection wavelength：285nm.
(5) replica test：Same batch of sample is taken, 6 parts of need testing solutions is prepared by step (2) method is parallel, enters respectively
Sample, calculates the relative retention time of shared fingerprint peakses and the RSD values of relative peak area, while being calculated with similarity evaluation software
The similarity of each chromatographic fingerprinting, as a result RSD values be respectively less than 3%, similarity is all higher than 0.990, shows this method reappearance
Well.
(6) stability test：
Take with a collection of need testing solution, respectively in 0h, 3h, 6h, 12h, 18h, 24h sample introductions calculate shared fingerprint peakses
The RSD values of relative retention time and relative peak area, while calculating the similar of each chromatographic fingerprinting with similarity evaluation software
Spend, as a result RSD values are respectively less than 3%, and similarity is all higher than 0.990, show that need testing solution is stable in 24 hours.
(7) replica test
Same batch Beijing enamel ware blood fat regulating capsule sample is taken, precision weighs 6 parts and is measured respectively, record chromatographic peak area, meter
The content of aurantiin in test sample, rutin, Chrysin and Galangin is calculated, as a result the average content of aurantiin is 22.76mg/g,
RSD is 0.72%, and the average content of rutin is 7.70mg/g, and RSD is 0.88%, and the average content of Chrysin is 2.45mg/g,
RSD is 0.87%, and the average content of Galangin is 5.18mg/g, and RSD is 0.74%, shows that the method reappearance is good.
(8) average recovery is tested
Take same batch known content (aurantiin 22.76mg/g, rutin 7.70mg/g, Chrysin 2.45mg/g and Gao Liang
Jiang Su 5.18mg/g) Beijing enamel ware blood fat regulating capsule sample (lot number：1311001) 9 parts, every part of about 0.125g is accurately weighed, respectively essence
It is close to add a certain amount of aurantiin, rutin, Chrysin and Galangin reference substance mixed solution 25mL, according under " 2.1.3 " item
Method prepares need testing solution, is measured according to chromatographic condition under " 2.1.1 " item, calculates aurantiin, rutin, Chrysin and height
The rate of recovery of alpinin.As a result aurantiin average recovery rate is that 98.49%, RSD is 1.09% (n=9), rutin average recovery rate
It is 1.89% (n=9) for 98.94%, RSD, Chrysin average recovery rate is that 99.15%, RSD is 1.33% (n=9), Gao Liang
Jiang Su average recovery rates are that 98.73%, RSD is 1.57% (n=9), show that this content assaying method has the good rate of recovery,
Method is reliable.
Beneficial effects of the present invention are：The inventive method is simple to operation, can be quickly and accurately to Beijing enamel ware blood fat regulating capsule
Quality be measured, precision, reproducible, the degree of accuracy is high, and the rate of recovery is good.
Embodiment
Embodiment：A kind of Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method of the present invention, it comprises the following steps：
(1) preparation of reference substance solution：Take aurantiin reference substance, control substance of Rutin, Chrysin reference substance and Galangin
Appropriate reference substance, it is accurately weighed, plus methanol be made every 1mL containing the μ g of aurantiin 80.23, the μ g of rutin 68.12, the μ g of Chrysin 22.82,
The μ g of Galangin 45.57 mixed solution, is produced；
(2) preparation of need testing solution：This product content about 0.25g is taken, is put in conical flask with cover, precision adds methanol
25mL, weighed weight ultrasonically treated 30 minutes, is let cool, then weighed weight, and the weight of less loss is supplied with methanol, is shaken up, filtration,
Subsequent filtrate is taken, is produced；
(3) determination method：Precision is drawn in reference substance solution and each 10 μ l of need testing solution, injection high performance liquid chromatograph,
It is measured according to setting chromatographic condition, records HPLC chromatogram；
(4) precision test：Take with a collection of need testing solution, continuous sample introduction 6 times, calculate 1~No. 30 shared fingerprint peaks
Relative retention time and relative peak area RSD values, while calculating the phase of each chromatographic fingerprinting with similarity evaluation software
Like degree.As a result RSD values are respectively less than 3%, and similarity is all higher than 0.990.
It is described set chromatographic condition as：
Chromatographic column：SHISEIDO Capcell pak MG C18 posts (4.6 × 250mm, 5 μm)；Mobile phase：Using methanol as
Mobile phase A, using 0.1% phosphate aqueous solution as Mobile phase B, gradient elution is carried out by table 2；Flow velocity：1.0mL/min；Column temperature：25
℃；Detection wavelength：285nm.
(5) replica test：Same batch of sample is taken, 6 parts of need testing solutions is prepared by step (2) method is parallel, enters respectively
Sample, calculates the relative retention time of shared fingerprint peakses and the RSD values of relative peak area, while being calculated with similarity evaluation software
The similarity of each chromatographic fingerprinting, as a result RSD values be respectively less than 3%, similarity is all higher than 0.990, shows this method reappearance
Well.
(6) stability test：
Take with a collection of need testing solution, respectively in 0h, 3h, 6h, 12h, 18h, 24h sample introductions calculate shared fingerprint peakses
The RSD values of relative retention time and relative peak area, while calculating the similar of each chromatographic fingerprinting with similarity evaluation software
Spend, as a result RSD values are respectively less than 3%, and similarity is all higher than 0.990, show that need testing solution is stable in 24 hours.
(7) replica test
Same batch Beijing enamel ware blood fat regulating capsule sample is taken, precision weighs 6 parts and is measured respectively, record chromatographic peak area, meter
The content of aurantiin in test sample, rutin, Chrysin and Galangin is calculated, as a result the average content of aurantiin is 22.76mg/g,
RSD is 0.72%, and the average content of rutin is 7.70mg/g, and RSD is 0.88%, and the average content of Chrysin is 2.45mg/g,
RSD is 0.87%, and the average content of Galangin is 5.18mg/g, and RSD is 0.74%, shows that the method reappearance is good.
(8) average recovery is tested
Take same batch known content (aurantiin 22.76mg/g, rutin 7.70mg/g, Chrysin 2.45mg/g and Gao Liang
Jiang Su 5.18mg/g) Beijing enamel ware blood fat regulating capsule sample (lot number：1311001) 9 parts, every part of about 0.125g is accurately weighed, respectively essence
It is close to add a certain amount of aurantiin, rutin, Chrysin and Galangin reference substance mixed solution 25mL, according under " 2.1.3 " item
Method prepares need testing solution, is measured according to chromatographic condition under " 2.1.1 " item, calculates aurantiin, rutin, Chrysin and height
The rate of recovery of alpinin.As a result aurantiin average recovery rate is that 98.49%, RSD is 1.09% (n=9), rutin average recovery rate
It is 1.89% (n=9) for 98.94%, RSD, Chrysin average recovery rate is that 99.15%, RSD is 1.33% (n=9), Gao Liang
Jiang Su average recovery rates are that 98.73%, RSD is 1.57% (n=9), show that this content assaying method has the good rate of recovery,
Method is reliable.
The inventive method is simple to operation, and quality that can be quickly and accurately to Beijing enamel ware blood fat regulating capsule is measured, accurate
Degree, reproducible, the degree of accuracy is high, and the rate of recovery is good.
Specifically,
1 instrument and material
1.1 instrument
Waters e2695-2998 types high performance liquid chromatographs (U.S.)；PAD detectors, quaternary gradient pump, Empower2
Data processing software system；KQ5200DE types supersonic extractors (Kunshan)；Mettler Toledo XS205DU electronic analysis day
Flat (Switzerland)；DHG-9203A types electric heating constant-temperature blowing drying box (Shanghai)；
HWS26 types electric-heated thermostatic water bath (Shanghai)；Millipore Advantage A10 ultrapure water systems (U.S.).
1.2 material
(17 batches, 1) lot number is shown in Table Beijing enamel ware blood fat regulating capsule, is provided by the second institute of traditional Chinese medicine of Guangdong Province Drug Manufacturing Room；Pummelo pee medicinal material
(lot number：20130504th, 20130602,20130605,20130802,20131009), ginkgo leaf medicinal material (lot number：20130502、
20130601st, 20130703,20130705,20130903), Gynostemma pentaphyllum (lot number：20130601、20130703、
20130801st, 20130903,20131002), propolis medicinal material (lot number：20130501、20130602、20130603、
20130701st, 20130801) it is purchased from Guangzhou north and south row prepared slices of Chinese crude drugs Co., Ltd；
Aurantiin reference substance (lot number：110722-200610), Kaempferide reference substance (lot number：110861-200808), Mongolian oak
Skin element reference substance (lot number：100081-200406), control substance of Rutin (lot number：100080-200707), Galangin reference substance
(lot number：111699-201001), Chrysin reference substance (lot number：111701-201001) Chinese food drug assay is purchased to grind
Study carefully institute.
The Beijing enamel ware blood fat regulating capsule sample lot number of table 1
Liquid phase is chromatographically pure (Merck) with methanol, acetonitrile；Liquid phase is ultra-pure water with water；Extraction is Guangdong Province second with water
Institute of traditional Chinese medicine Drug Manufacturing Room make purified water by oneself；Extraction is that analysis is pure with reagents such as methanol, glacial acetic acid, formic acid, phosphoric acid.
2 methods and result
2.1 Beijing enamel ware blood fat regulating capsule fingerprint spectrum methods and result
2.1.1, chromatographic condition
Chromatographic column：SHISEIDO Capcell pak MG C18 posts (4.6 × 250mm, 5 μm)；Mobile phase：Using methanol as
Mobile phase A, using 0.1% phosphate aqueous solution as Mobile phase B, gradient elution is carried out by table 2；Flow velocity：1.0mL/min；Column temperature：25
℃；Detection wavelength：285nm.
The condition of gradient elution of table 2
2.1.2 the preparation of reference substance solution
Take aurantiin reference substance, control substance of Rutin, Chrysin reference substance and Galangin reference substance appropriate, it is accurately weighed,
Plus methanol be made every 1mL containing the μ g of aurantiin 80.23, the μ g of rutin 68.12, the μ g of Chrysin 22.82, the μ g of Galangin 45.57 it is mixed
Solution is closed, is produced.
2.1.3 the preparation of need testing solution
This product content about 0.25g is taken, is put in conical flask with cover, precision adds methanol 25mL, weighed weight is ultrasonically treated
30 minutes, let cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, is filtered, is taken subsequent filtrate, produce.
2.1.4 determination method
Precision is drawn in reference substance solution and each 10 μ l of need testing solution, injection high performance liquid chromatograph, according to above-mentioned color
Spectral condition is measured, and records HPLC chromatogram.
2.1.5 methodological study
2.1.5.1 precision test
Take with a collection of need testing solution (lot number：1311001), continuous sample introduction 6 times, using No. 23 peak Chrysins as reference peak,
The relative retention time of 1~No. 30 shared fingerprint peaks and the RSD values of relative peak area are calculated, while using similarity evaluation software
Calculate the similarity of each chromatographic fingerprinting.As a result RSD values are respectively less than 3%, and similarity is all higher than 0.990, shows instrument precision
Degree is good.
2.1.5.2 replica test
Take same batch of sample (lot number：1311001), precision weighs 6 parts respectively, according to legal system available below " 2.1.3 " item
Test sample solution, sample introduction respectively, using No. 23 peak Chrysins for reference peak, when calculating the relative reservation of 1~No. 30 shared fingerprint peaks
Between and relative peak area RSD values, while with the similarity of each chromatographic fingerprinting of similarity evaluation software calculating.As a result RSD
Value is respectively less than 3%, and similarity is all higher than 0.990, shows that this method reappearance is good.
2.1.5.3 stability test
Take with a collection of need testing solution (lot number：1311001), respectively in 0h, 3h, 6h, 12h, 18h, 24h sample introductions, with 23
Number peak Chrysin is the RSD values of relative retention time and relative peak area that 1~No. 30 shared fingerprint peaks are calculated with reference to peak,
The similarity of each chromatographic fingerprinting is calculated with similarity evaluation software simultaneously.As a result RSD values are respectively less than 3%, and similarity is big
In 0.990, show that need testing solution is stable in 24 hours.
2.1.6 the foundation of finger-print and similarity analysis
The Beijing enamel ware blood fat regulating capsule of different batches is taken, according to legal system available test sample solution below " 2.1.3 " item, according to
Chromatographic condition is analyzed under " 2.1.1 " item, records HPLC collection of illustrative plates, and 17 batches of Beijing enamel ware blood fat regulating capsule finger-prints are imported into " Chinese medicine
Chromatographic fingerprinting similarity evaluation system (2004A editions) ", carries out chromatogram peak match, it is determined that 30 chromatographic peaks are shared spy
Levy peak.By the comparison with reference substance chromatogram, 4 chromatographic peaks are pointed out, wherein No. 9 peaks are aurantiin, No. 11 peaks are rutin,
No. 23 peaks are Chrysin, and No. 24 peaks are Galangin, and 17 batch Beijing enamel ware blood fat regulating capsule similarities the results are shown in Table more than 0.9
3。
The Beijing enamel ware blood fat regulating capsule fingerprint similarity result of table 3
2.1.7 finished product is investigated with raw medicinal material correlation
The Beijing enamel ware blood fat regulating capsule finished product and raw medicinal material of corresponding lot number are taken, according to legal system available test product below " 2.1.3 " item
Solution, is measured according to chromatographic condition under " 2.1.1 " item, obtains same batch Beijing enamel ware blood fat regulating capsule finished product, Exocarpium Citri Grandis, ginkgo
The efficient liquid-phase chromatograph finger print atlas of leaf, gynostemma pentaphylla and propolis.As a result show, belonging to the chromatographic peak of Exocarpium Citri Grandis there are 4, respectively
For No. 8, No. 9, No. 10 and No. 14 chromatographic peaks；Belonging to the chromatographic peak of ginkgo leaf has 2, respectively No. 3 and No. 5 chromatographic peaks；Return
Belonging to the chromatographic peak of gynostemma pentaphylla has 1, is No. 18 chromatographic peaks；Belonging to the chromatographic peak of propolis has 12, respectively No. 7, No. 16,
No. 19, No. 20, No. 21, No. 22, No. 23, No. 24, No. 27, No. 28, No. 29 and No. 30 chromatographic peaks.
2.2 the finger-print multi objective quantitative analysis method of Beijing enamel ware blood fat regulating capsule and result
2.2.1 chromatographic condition
With chromatographic condition under " 2.1.1 " item.
2.2.2 the preparation of reference substance solution
Take aurantiin reference substance, control substance of Rutin, Chrysin reference substance and Galangin reference substance appropriate, it is accurately weighed,
Plus every 1mL is made containing the μ g of aurantiin 321.20, the μ g of rutin 119.10, the μ g of Chrysin 45.70, Galangin 74.20 μ g in methanol
Mixed solution, is produced, standby.
2.2.3 the preparation of need testing solution
With need testing solution preparation method under " 2.1.3 " item.
2.2.4 methodological study
2.2.4.1 linear relationship is investigated
Precision measures mixed reference substance solution 1mL, 2mL, 4mL, 6mL, 8mL, the 10mL prepared under " 2.2.2 " item, point
Do not put in 10mL measuring bottles, add methanol dilution to scale, every 1mL μ g, 64.24 μ g, 128.48 μ g containing aurantiin 32.12 are made,
192.72 μ g, 256.96 μ g, 321.20 μ g；The μ of the μ of rutin 11.91 μ g, 23.82 μ g, 47.64 g, 71.46 μ g, 95.28 g, 119.10
μg；Chrysin 4.57 μ g, 9.14 μ g, 18.28 μ g, 27.42 μ g, 36.56 μ g, 45.70 μ g and Galangin 7.42 μ g, 14.84 μ
G, 29.68 μ g, 44.52 μ g, 59.36 μ g, 74.20 μ g solution.Precision draws above-mentioned each 10 μ L of mixed reference substance solution, injection
Liquid chromatograph, is measured according to chromatographic condition under " 2.1.1 " item.Using peak area as ordinate (Y), it is with sample size (ng)
Abscissa (X) carries out linear regression, obtains regression equation Y aurantiins=1472.33X-439.01, r=0.99991；Y rutins=
415.85X+58.99, r=0.99990；Y Chrysins=4585.29X-881.90, r=0.99991 and Y Galangins=
817.36X-65.23, r=0.99993.As a result show aurantiin in the range of 32.12~321.20ng, rutin 11.91~
In the range of 119.10ng, Chrysin is in the range of 4.57~45.70ng and Galangin is in the range of 7.42~74.20ng, its
Sample size (ng) is in good linear relation with peak area.It the results are shown in Table 4.
The linear relationship result of the test of table 4
2.2.4.2 precision test
Precision draws same mixed reference substance solution (the μ g/mL of aurantiin 192.72, the μ g/mL of rutin 71.46, Chrysin
27.42 μ g/mL, the μ g/mL of Galangin 44.52) 10 μ l, inject high performance liquid chromatograph, according to chromatographic condition under " 2.1.1 " item
Repeat sample introduction 6 times, record chromatogram.It is 0.66% to calculate aurantiin peak area value RSD, and rutin peak area value RSD is 0.47%,
Chrysin peak area value RSD is 0.42%, and Galangin area value RSD is 0.51%, shows that instrument precision is good, as a result sees
Table 5.
The instrument precision result of the test of table 5
2.2.4.3 stability test
Precision draws same need testing solution (lot number：1311001) 10 μ l, respectively at 0h, 3h, 6h, 12h, 18h, 24h enters
Sample is determined.It is 0.21% to calculate aurantiin peak area value RSD, and rutin peak area value RSD is 0.46%, Chrysin peak area value
RSD is 0.51%, and Galangin area value RSD is 0.35%.Show that need testing solution is good in 24h internal stabilities, as a result tie
Fruit is shown in Table 6.
The test sample stability test result of table 6
2.2.4.4 replica test
Take same batch Beijing enamel ware blood fat regulating capsule sample (lot number：1311001), precision weighs 6 parts respectively, according to " 2.1.3 "
Legal system available test sample solution below, is measured according to chromatographic condition under " 2.1.1 " item, records chromatographic peak area, calculates and supplies
Aurantiin in test product, rutin, the content of Chrysin and Galangin.As a result the average content of aurantiin is 22.76mg/g, RSD
For 0.72%, the average content of rutin is 7.70mg/g, and RSD is 0.88%, and the average content of Chrysin is 2.45mg/g, RSD
For 0.87%, the average content of Galangin is 5.18mg/g, and RSD is 0.74%, shows that the method reappearance is good, as a result sees
Table 7.
The replica test result of table 7
2.2.4.5 average recovery is tested
Take same batch known content (aurantiin 22.76mg/g, rutin 7.70mg/g, Chrysin 2.45mg/g and Gao Liang
Jiang Su 5.18mg/g) Beijing enamel ware blood fat regulating capsule sample (lot number：1311001) 9 parts, every part of about 0.125g is accurately weighed, respectively essence
It is close to add a certain amount of aurantiin, rutin, Chrysin and Galangin reference substance mixed solution 25mL, according under " 2.1.3 " item
Method prepares need testing solution, is measured according to chromatographic condition under " 2.1.1 " item, calculates aurantiin, rutin, Chrysin and height
The rate of recovery of alpinin.As a result aurantiin average recovery rate is that 98.49%, RSD is 1.09% (n=9), rutin average recovery rate
It is 1.89% (n=9) for 98.94%, RSD, Chrysin average recovery rate is that 99.15%, RSD is 1.33% (n=9), Gao Liang
Jiang Su average recovery rates are that 98.73%, RSD is 1.57% (n=9), show that this content assaying method has the good rate of recovery,
Method is reliable, the results are shown in Table 8~table 11.
The aurantiin average recovery result of the test of table 8
The rutin average recovery result of the test of table 9
The Chrysin average recovery result of the test of table 10
The Galangin average recovery result of the test of table 11
2.2.5 the assay result of sample
Different batches Beijing enamel ware blood fat regulating capsule sample is taken, according to legal system available test sample solution below " 2.1.3 " item, according to
Chromatographic condition carries out assay under " 2.1.1 " item, calculates the content of aurantiin, rutin, Chrysin and Galangin, is shown in Table
12。
12 each batches of Beijing enamel ware blood fat regulating capsule sample size measurement results of table
As a result show, in different batches Beijing enamel ware blood fat regulating capsule 4 indexs of aurantiin, rutin, Chrysin and Galangin into
There is difference in the content divided.Naringin content is between 8.18~23.11mg/g；Rutin content 3.11~8.24mg/g it
Between；White poplar cellulose content is between 1.56~2.73mg/g；Galangin content is between 0.82~5.29mg/g.《Beijing enamel ware adjusts fat
Capsule pharmaceutical standard》The content of aurantiin (C27H32O14) is provided, 14.0mg/g must not be less than；Chrysin (C15H10O4)
Content, must not be less than 1.40mg/g；The content of Galangin (C15H10O5), must not be less than 0.60mg/g, and the content of rutin
Do not provide temporarily, according to this experiment acquired results, the content of tentative rutin (C27H30O16) must not be less than 5.45mg/g.
The above described is only a preferred embodiment of the present invention, not making any formal limitation to the present invention.Appoint
What those skilled in the art, without departing from the scope of the technical proposal of the invention, all using the side of the disclosure above
Method and technology contents make many possible variations and modification to technical solution of the present invention, or are revised as the equivalent reality of equivalent variations
Apply example.Therefore every content without departing from technical solution of the present invention, the equivalent change that shape, construction and principle according to the present invention are made
Change, all should be covered by protection scope of the present invention.
Claims (6)
1. a kind of Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method, it is characterised in that it comprises the following steps：
(1) preparation of reference substance solution：Take aurantiin reference substance, control substance of Rutin, Chrysin reference substance and Galangin control
Appropriate product, it is accurately weighed, plus every 1mL is made containing the μ g of aurantiin 80.23, the μ g of rutin 68.12, Chrysin 22.82 μ g, Gao Liang in methanol
The μ g of Jiang Su 45.57 mixed solution, is produced；
(2) preparation of need testing solution：This product content about 0.25g is taken, is put in conical flask with cover, precision adds methanol 25mL,
Weighed weight, ultrasonically treated 30 minutes, is let cool, then weighed weight, and the weight of less loss is supplied with methanol, is shaken up, filtration, takes continuous filter
Liquid, is produced；
(3) determination method：Precision is drawn in reference substance solution and each 10 μ l of need testing solution, injection high performance liquid chromatograph, according to
Setting chromatographic condition is measured, and records HPLC chromatogram；
(4) precision test：Take with a collection of need testing solution, continuous sample introduction 6 times, calculate the phase of 1~No. 30 shared fingerprint peaks
To retention time and the RSD values of relative peak area, while calculating the similarity of each chromatographic fingerprinting with similarity evaluation software.
As a result RSD values are respectively less than 3%, and similarity is all higher than 0.990.
2. Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method according to claim 1, it is characterised in that the setting color
Spectral condition is：
Chromatographic column：SHISEIDO Capcell pak MG C18 posts (4.6 × 250mm, 5 μm)；Mobile phase：Using methanol as flowing
Phase A, using 0.1% phosphate aqueous solution as Mobile phase B, gradient elution is carried out by table 2；Flow velocity：1.0mL/min；Column temperature：25℃；Inspection
Survey wavelength：285nm.
3. Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method according to claim 1, it is characterised in that its also include with
Lower step：
(5) replica test：Same batch of sample is taken, 6 parts of need testing solutions are prepared by step (2) method is parallel, respectively sample introduction, meter
The relative retention time of shared fingerprint peakses and the RSD values of relative peak area are calculated, while calculating assorted with similarity evaluation software
The similarity of finger-print is composed, as a result RSD values are respectively less than 3%, and similarity is all higher than 0.990, shows that this method reappearance is good.
4. Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method according to claim 1, it is characterised in that its also include with
Lower step：
(6) stability test：
Take with a collection of need testing solution, respectively in 0h, 3h, 6h, 12h, 18h, 24h sample introductions calculate the relative of shared fingerprint peakses
The RSD values of retention time and relative peak area, while the similarity of each chromatographic fingerprinting is calculated with similarity evaluation software, knot
Fruit RSD values are respectively less than 3%, and similarity is all higher than 0.990, show that need testing solution is stable in 24 hours.
5. Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method according to claim 1, it is characterised in that its also include with
Lower step：
(7) replica test
Same batch Beijing enamel ware blood fat regulating capsule sample is taken, precision weighs 6 parts and is measured respectively, record chromatographic peak area, calculated and supply
Aurantiin in test product, rutin, the content of Chrysin and Galangin, as a result the average content of aurantiin is 22.76mg/g, RSD
For 0.72%, the average content of rutin is 7.70mg/g, and RSD is 0.88%, and the average content of Chrysin is 2.45mg/g, RSD
For 0.87%, the average content of Galangin is 5.18mg/g, and RSD is 0.74%, shows that the method reappearance is good.
6. Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method according to claim 1, it is characterised in that its also include with
Lower step：
(8) average recovery is tested
Take same batch known content (aurantiin 22.76mg/g, rutin 7.70mg/g, Chrysin 2.45mg/g and Galangin
Beijing enamel ware blood fat regulating capsule sample (lot number 5.18mg/g)：1311001) 9 parts, every part of about 0.125g is accurately weighed, accurate respectively to add
Enter a certain amount of aurantiin, rutin, Chrysin and Galangin reference substance mixed solution 25mL, according to method under " 2.1.3 " item
Need testing solution is prepared, is measured according to chromatographic condition under " 2.1.1 " item, aurantiin, rutin, Chrysin and galangal is calculated
The rate of recovery of element.As a result aurantiin average recovery rate is that 98.49%, RSD is 1.09% (n=9), and rutin average recovery rate is
98.94%, RSD are 1.89% (n=9), and Chrysin average recovery rate is that 99.15%, RSD is 1.33% (n=9), galangal
Plain average recovery rate is that 98.73%, RSD is 1.57% (n=9), shows that this content assaying method has the good rate of recovery, side
Method is reliable.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233582.1A CN107064352A (en) | 2017-04-11 | 2017-04-11 | Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710233582.1A CN107064352A (en) | 2017-04-11 | 2017-04-11 | Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107064352A true CN107064352A (en) | 2017-08-18 |
Family
ID=59602262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710233582.1A Pending CN107064352A (en) | 2017-04-11 | 2017-04-11 | Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107064352A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107741466A (en) * | 2017-11-28 | 2018-02-27 | 深圳海王医药科技研究院有限公司 | Method that is a kind of while determining rutin and naringin content |
| CN112345655A (en) * | 2019-08-06 | 2021-02-09 | 大理大学 | Establishing method of wasp venom fingerprint, wasp venom fingerprint and application of wasp venom fingerprint |
-
2017
- 2017-04-11 CN CN201710233582.1A patent/CN107064352A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107741466A (en) * | 2017-11-28 | 2018-02-27 | 深圳海王医药科技研究院有限公司 | Method that is a kind of while determining rutin and naringin content |
| CN112345655A (en) * | 2019-08-06 | 2021-02-09 | 大理大学 | Establishing method of wasp venom fingerprint, wasp venom fingerprint and application of wasp venom fingerprint |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106198782B (en) | A kind of method of quality control of the content analysis that 18 components can be achieved at the same time in Ilex Latifolia Thunb and similarity evaluation | |
| CN105486771B (en) | The fingerprint atlas detection method of Xiao Chai Hu granules compound preparation | |
| CN106841478A (en) | It is a kind of at the same determine chrysanthemum indicum in seven kinds of methods of the content of composition | |
| CN109596751A (en) | A kind of Mailuo Ning oral liquid component detection method of clearing heat and nourishing yin promoting blood circulation | |
| CN103149310B (en) | Fingerprint building method of Shenxiong glucose injection preparation | |
| CN109521103B (en) | Method for detecting quality of oral liquid of Sanle pulp with qualitative and quantitative evaluation | |
| CN104483405B (en) | The detection method of the related substances in autonomic drug extract lamp-dish flower acetic | |
| CN107037146A (en) | A kind of method of the wooden chicken granular mass control of compound | |
| CN111487344B (en) | Method for detecting fingerprint spectrum of motherwort particles | |
| CN108459090B (en) | Quality control method of Jichuan decoction composition | |
| CN107064352A (en) | Beijing enamel ware blood fat regulating capsule fingerprint pattern quality control method | |
| CN110487925A (en) | A kind of method of chemical component in quick identification lilii and Rehmanniae Decoction | |
| CN110806457B (en) | Detection method of fingerprint of Sijun manna drink | |
| CN102048906B (en) | Content measurement method of abrus herb capsules | |
| CN108490083A (en) | The quality determining method of SuHuang ZhiKe capsule | |
| CN102707006B (en) | Quality detection method of cudrania tricuspidata formula granules | |
| CN102397331A (en) | Preparation and quality detection methods of high-purity honeysuckle flower-baikal skullcap root soluble powder | |
| CN110441445A (en) | A kind of construction method and flavones ingredient assay of cockscomb HPLC characteristic fingerprint pattern | |
| CN111505156B (en) | Fingerprint spectrogram quality determination method for herba Cirsii formulation granules | |
| CN108982738A (en) | The intermediate characteristic spectrum of zhenqi fuzheng granules or the construction method and detection method of zhenqi fuzheng granules finger-print | |
| CN112697899B (en) | Detection method of ginkgo flavonol glycosides | |
| CN111189952B (en) | Method for measuring and controlling specific content and quality of verbena | |
| CN102068599B (en) | Detection method for phlegm eliminating and asthma relieving cough syrup | |
| CN102759582B (en) | Method for determining flavonoids components of ophiopogon japonicus | |
| CN110412162A (en) | The quality determining method of the construction method of the HPLC characteristic spectrum of white perfume capsule and white fragrant capsule |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170818 |
|
| WD01 | Invention patent application deemed withdrawn after publication |