CN100573138C - The foundation of Xia sang ju formulation finger-print and finger-print - Google Patents
The foundation of Xia sang ju formulation finger-print and finger-print Download PDFInfo
- Publication number
- CN100573138C CN100573138C CNB2007100263469A CN200710026346A CN100573138C CN 100573138 C CN100573138 C CN 100573138C CN B2007100263469 A CNB2007100263469 A CN B2007100263469A CN 200710026346 A CN200710026346 A CN 200710026346A CN 100573138 C CN100573138 C CN 100573138C
- Authority
- CN
- China
- Prior art keywords
- rsd
- peaks
- mobile phase
- peak area
- retention time
- 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.)
- Active
Links
Images
Landscapes
- Medicinal Preparation (AREA)
Abstract
The foundation of Xia sang ju formulation finger-print and finger-print relate to the method for quality control of Chinese medicine preparation, specifically are to set up the HPLC finger-print to detect the method for alcohol extract in the Xia sang ju formulation.Method comprises: (a) preparation of reference substance solution; (b) preparation of need testing solution; (c) chromatographic condition: chromatographic column is filler with the octadecylsilane chemically bonded silica; Adopt gradient elution, moving phase is the gradient eluent that 0.1～3% acetic acid and methyl alcohol are formed; Column temperature: 25～50 ℃; The ultraviolet detection wavelength is 285～295nm; Flow velocity: 0.5～1.5mLmin
-1Time: 30～80min; (d) measure: high performance liquid chromatography gets finger-print.Beneficial effect of the present invention: can effectively characterize the quality of Xia sang ju formulation, help the quality of monitoring product; Have that method is easy, stable, precision is high, favorable reproducibility; Can differentiate the true and false quality of product quickly and accurately.
Description
Technical field
The present invention relates to the method for quality control of Chinese medicine preparation, be specifically related to the construction method and the standard finger-print of Xia sang ju formulation high performance liquid chromatography finger-print.
Background technology
Xia sang ju formulation is a raw material by Chinese crude drug selfheal, mulberry leaf, mother chrysanthemum, after water extracts, pharmaceutical technology is routinely made, have clear liver and improve vision, wind-dispelling heat-dissipating, the function removing arthritis with fixed pain caused by dampness, separate sore, be used for diseases such as anemopyretic cold, hot eyes headache, hypertension, dizziness and tinnitus, abscess of throat, treatment pyogenic infections from tumour or sore.Xia sang ju formulation has recorded the mulberry and chrysanthemum granules agent in the 15 in Drug Standard of Ministry of Public Health of the Peoples Republic of China Chinese traditional patent formulation preparation, is well received Chinese medicine preparation, now existing summer Sang Ju Yin sugar free granule, summer mulberry-chrysanthemum oral liquid, the listing of formulations such as summer Sang Ju Yin beverage.
Country and enterprise mainly adopt thin-layered chromatography to differentiate the composition of the ursolic acid whether the main ingredient selfheal is arranged to the quality control of mulberry and chrysanthemum granules;
Yang Dongmei etc. are at document " quality standard research of xiasangju instant herbal medicine " Anhui medicine, and reported with in spectrophotometry Xia sang ju formulation content of total flavone at 2001,5 (3): 218;
Ma Wen etc. disclose the analytical approach that detects ursolic acid in the summer Sang Ju Yin with the high performance liquid chromatography UV, visible light at magazine " Journal of Analytical Science " 2004,20 (3): 284; Huang Shuzhang has reported that at document " modern Chinese herbal medicine research with put into practice " 2004,18 (3): 33 employing HPLC method measured content of ursolic acid in the mulberry and chrysanthemum granules; The state-run grade of bear is at document " Jiangxi College of Traditional Chinese Medicine journal " 2004,16 (6): reported 48) with the HPLC method and measured contained active component chlorogenic acid contents method in the particle.
Below all be to differentiate or the method for assay, be difficult to characterize the physicochemical characteristic of Xia sang ju formulation comprehensively, therefore, remain further perfect the method for quality control of compound Xia sang ju formulation at indivedual compositions.
Summary of the invention
Purpose of the present invention provides a kind of new method for the quality control and the real and fake discrimination of Xia sang ju formulation, and by setting up the method for Xia sang ju formulation finger-print, and method obtains the finger-print of Xia sang ju formulation common pattern thus.
The technical scheme that is adopted to achieve the object of the present invention: set up the method for Xia sang ju formulation finger-print with high performance liquid chromatography, specifically comprise the steps:
(a) preparation of reference substance solution: precision takes by weighing the chlorogenic acid reference substance, is mixed with the solution that every 1ml contains chlorogenic acid 0.2～0.5mg with methyl alcohol, in contrast product solution;
(b) preparation of need testing solution: the A. precision takes by weighing that mulberry and chrysanthemum granules agent powder is dissolved in the methyl alcohol, after the sonicated, filter, and filtrate volatilizes, and the residue dissolve with methanol is put volumetric flask, and molten calmly, membrane filtration promptly gets need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid, volatilizes, and the residue dissolve with methanol is put volumetric flask, and fixed molten, membrane filtration promptly gets need testing solution;
(c) chromatographic condition: chromatographic column is filler with the octadecylsilane chemically bonded silica; Adopt gradient elution, moving phase is the gradient eluent that acetic acid and methyl alcohol are formed; Ultraviolet detection: wavelength is 285～295nm;
(d) measure: the accurate need testing solution of drawing injects high performance liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
Further preferable methods can be implemented by following step:
(a) preparation of reference substance solution: precision takes by weighing the chlorogenic acid reference substance and places volumetric flask in right amount, is diluted to scale with methyl alcohol, is mixed with the solution that every 1ml contains chlorogenic acid 0.2～0.5mg, in contrast product solution;
(b) preparation of need testing solution: the A. precision takes by weighing mulberry and chrysanthemum granules agent powder 0.1～20g and puts in 20～100mL ground Erlenmeyer flask, accurate methyl alcohol 20～100ml, the jam-pack of adding, after immersion, the sonicated, supply the weight that subtracts mistake, shake up with methyl alcohol, filter, filtrate volatilizes, and the residue dissolve with methanol is put 1～5mL volumetric flask, add methyl alcohol to scale, shake up, membrane filtration promptly gets need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid 25～50mL, volatilizes in the water-bath, and the residue dissolve with methanol is put 1～5mL volumetric flask, adds methyl alcohol to scale, shakes up, and membrane filtration promptly gets need testing solution;
(c) chromatographic condition: chromatographic column is filler with the octadecylsilane chemically bonded silica; Adopt gradient elution, moving phase is the gradient eluent that 0.1～3% acetic acid and methyl alcohol are formed; Column temperature: 25～50 ℃; Ultraviolet detection: wavelength is 285～295nm; Flow velocity: 0.5～1.5mLmin
-1Time: 30～80min;
(d) measure: accurate need testing solution 5～25 μ L that draw inject high performance liquid chromatograph, according to high effective liquid chromatography for measuring, obtain finger-print.
The detection method of the best of the present invention can be implemented by following step: (b) preparation of need testing solution: the A. precision takes by weighing mulberry and chrysanthemum granules agent powder 5.0g and puts in the 50mL ground Erlenmeyer flask, accurate methyl alcohol 50ml, the jam-pack of adding, soaked 2 hours, sonicated was put to room temperature after 30 minutes, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, filtrate volatilizes, and the residue dissolve with methanol is put the 5mL volumetric flask, add methyl alcohol to scale, shake up, use membrane filtration, promptly get need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid 25mL, volatilizes in the water-bath, and the residue dissolve with methanol is put the 5mL volumetric flask, adds methyl alcohol to scale, shakes up, and membrane filtration promptly gets need testing solution; (c) chromatographic condition: chromatographic column is filler with the octadecylsilane chemically bonded silica; Moving phase is the gradient eluent that 1% acetic acid and methyl alcohol are formed, column temperature: 30 ℃; The detection wavelength is 290nm; Flow velocity 1.0mL.min
-1Analysis time: 65min.
In step (c), described gradient elution, the preferred gradient elution program carries out with following volumetric concentration configuration: mobile phase A is 1% acetum, and Mobile phase B is a methyl alcohol.
In the time of 0 minute, mobile phase A is 100%, Mobile phase B is 0%;
In the time of 5 minutes, mobile phase A is 93%, Mobile phase B is 7%;
In the time of 9 minutes, mobile phase A is 90%, Mobile phase B is 10%;
In the time of 13 minutes, mobile phase A is 82%, Mobile phase B is 18%;
In the time of 20 minutes, mobile phase A is 78%, Mobile phase B is 22%;
In the time of 35 minutes, mobile phase A is 60%, Mobile phase B is 40%;
In the time of 55 minutes, mobile phase A is 30%, Mobile phase B is 70%;
In the time of 60 minutes, mobile phase A is 0%, Mobile phase B is 100%;
In the time of 65 minutes, mobile phase A is 100%, Mobile phase B is 0%;
Specifically see the following form:
| Moving phase | Time | Flow mL.min -1 | A 1% sodium acetate mL | B methyl alcohol mL |
| 1 | 0 | 1 | 100 | 0 |
| 2 | 5 | 1 | 93 | 7 |
| 3 | 9 | 1 | 90 | 10 |
| 4 | 13 | 1 | 82 | 18 |
| 5 | 20 | 1 | 78 | 22 |
| 6 | 35 | 1 | 60 | 40 |
| 7 | 55 | 1 | 30 | 70 |
| 8 | 60 | 1 | 0 | 100 |
| 9 | 65 | 1 | 100 | 0 |
There are 20 features to have the peak in the finger-print according to the method for building up gained Xia sang ju formulation of finger-print provided by the present invention, wherein No. 8 chromatographic peaks are that chlorogenic acid is with reference to the peak, highest peak is 16 chromatographic peaks, the collection of illustrative plates total length is 65min, the peak of the super total peak area 1% of wherein unimodal area has 20, and they are 1～No. 20 peaks; The peak of the super total peak area 5% of wherein unimodal area has 4, and they are No. 4 peaks, No. 8 peaks (chlorogenic acid), No. 16 peaks, No. 20 peaks; The peak of the super total peak area 10% of unimodal area has 1, is No. 16 peaks.Specific as follows:
No. 1 peak, average retention time RT is 1.57min, and RSD is 2.06%, and peak area is 1542541, and RSD is 23.75%.
No. 2 peaks, average retention time RT is 1.91min, and RSD is 1.08%, and peak area is 1660403, and RSD is 23.47%.
No. 3 peaks, average retention time RT is 3.56min, and RSD is 1.43%, and peak area is 1572609, and RSD is 24.14%.
No. 4 peaks, average retention time RT is 10.83min, and RSD is 0.55%, and peak area is 3730363, and RSD is 23.20%.
No. 5 peaks, average retention time RT is 11.22min, and RSD is 0.63%, and peak area is 1023133, and RSD is 18.56%.
No. 6 peaks, average retention time RT is 14.60min, and RSD is 0.48%, and peak area is 763899, and RSD is 33.34%.
No. 7 peaks, average retention time RT is 15.25min, and RSD is 0.66%, and peak area is 2077260, and RSD is 24.81%.
No. 8 peaks are chlorogenic acid, and average retention time RT is 19.85min, and RSD is %0.35, and peak area is 2613097, and RSD16.46 is %.
No. 9 peaks, average retention time RT is 20.69min, and RSD is 0.78%, and peak area is 805465, and RSD is 18.00%.
No. 10 peaks, average retention time RT is 22.66min, and RSD is 0.51%, and peak area is 2258797, and RSD is 13.57%.
No. 11 peaks, average retention time RT is 35.66min, and RSD is %1.02, and peak area is 588056, and RSD is 28.63%.
No. 12 peaks, average retention time RT is 36.58min, and RSD is 0.64%, and peak area is 1739577, and RSD is 23.95%.
No. 13 peaks, average retention time RT is 37.48min, and RSD is 0.87%, and peak area is 1144232, and RSD is 29.03%.
No. 14 peaks, average retention time RT is 37.99min, and RSD is 0.61%, and peak area is 1210576, and RSD is 24.05%.
No. 15 peaks, average retention time RT is 39.32min, and RSD is 0.72%, and peak area is 838066, and RSD is 20.36%.
No. 16 peaks, average retention time RT is 40.45min, and RSD is 0.85%, and peak area is 18321566, and RSD is 17.22%.
No. 17 peaks, average retention time RT is 44.58min, and RSD is 0.91%, and peak area is 1925024, and RSD is 20.23%.
No. 18 peaks, average retention time RT is 45.45min, and RSD is 1.13%, and peak area is 1666840, and RSD is 31.43%.
No. 19 peaks, average retention time RT is 48.01min, and RSD is 0.88%, and peak area is 550287, and RSD is 22.56%.
No. 20 peaks, average retention time RT is 55.14min, RSD is 0.77%.Peak area is 3131146, and RSD is 27.56%.
RT is that the peak that 60min occurs later on is a solvent peak.
Principle of the present invention is to be the extract of medicinal material selfheal, mulberry leaf, mother chrysanthemum according to active component main in the Xia sang ju formulation, its finger-print can be held the quality of Xia sang ju formulation from the whole looks of chromatogram, and said Xia sang ju formulation comprises that solid pharmaceutical preparation and liquid preparation such as summer Sang Ju Yin contain sugared granule, sugar free granule, tablet, capsule, beverage, mixture and oral liquid etc.The Xia sang ju formulation of measuring by the national standard preparation with this method all can obtain identical, akin finger-print.
Beneficial effect of the present invention is as follows:
(1) the HPLC finger-print of setting up with method provided by the present invention can characterize the quality of Xia sang ju formulation effectively, helps the quality of overall monitor product.
(2) finger-print is paid attention to each front and back that constitute fingerprint characteristic peak order and mutual relationship, pay attention to whole facial feature, both avoided judging the one-sidedness of mulberry and chrysanthemum granules agent total quality that having reduced again was the possibility of the artificial processing of requisite quality because of measuring one, two chemical constitution.
(3) the present invention has that method is easy, stable, precision is high, high repeatability and other advantages.
(4) this method can be differentiated the true and false quality of product quickly and accurately.
Elaborate technical scheme of the present invention below by embodiment and accompanying drawing thereof, cited embodiment and accompanying drawing are to not restriction of the present invention.
Description of drawings
The HPLC finger-print of Fig. 1, mulberry and chrysanthemum granules agent
10 batches of original fingerprint collection of illustrative plates of Fig. 2, mulberry and chrysanthemum granules agent
Finger-print after 10 batches of couplings of Fig. 3, mulberry and chrysanthemum granules agent
The finger-print of the HPLC common pattern of Fig. 4, Xia sang ju formulation
Elaborate technical scheme of the present invention below by embodiment and accompanying drawing thereof, cited embodiment and accompanying drawing are to not restriction of the present invention.
Embodiment
Embodiment 1. detects the finger-print that the summer Sang Ju Yin contains sugared granule
1. instrument and reagent
1.1 instrument Waters2695 high performance liquid chromatograph, Waters 2996 UV-detector, Empower chromatographic work station, AE-200 type electronic analytical balance.
1.2 reagent chlorogenic acid reference substance; Methyl alcohol (chromatographically pure), secondary redistilled water, acetic acid (analyzing pure).
2. high performance liquid chromatography
2.1 chromatographic condition: chromatographic condition: chromatographic column is filler (SunfrieC with the octadecylsilane chemically bonded silica
18, analytical column 150mm * 4.6mm, 5um); Moving phase is the gradient eluent that 1% acetic acid and methyl alcohol are formed, column temperature: 30 ℃; Ultraviolet detection: wavelength is 290nm; Flow velocity 1.0m L.min
-1Analysis time: 65min.Sample size: each 10 μ L of need testing solution and reference substance solution.Theoretical cam curve is calculated by chlorogenic acid should be not less than 3000.
Gradient elution program such as following table:
| Moving phase | Time | Flow mL.min -1 | A 1% sodium acetate mL | B methyl alcohol mL |
| 1 | 0 | 1 | 100 | 0 |
| 2 | 5 | 1 | 93 | 7 |
| 3 | 9 | 1 | 90 | 10 |
| 4 | 13 | 1 | 82 | 18 |
| 5 | 20 | 1 | 78 | 22 |
| 6 | 35 | 1 | 60 | 40 |
| 7 | 55 | 1 | 30 | 70 |
| 8 | 60 | 1 | 0 | 100 |
| 9 | 65 | 1 | 100 | 0 |
2.2 the preparation of reference substance solution: precision takes by weighing chlorogenic acid reference substance 5.0mg and places the 25mL volumetric flask, is diluted to scale with methyl alcohol, promptly.
2.3 the preparation of need testing solution
The summer Sang Ju Yin contains the preparation of sugared granule need testing solution: precision takes by weighing mulberry and chrysanthemum granules 5.0g and puts in the tool plug triangular flask, accurate methyl alcohol 50ml, the jam-pack of adding, soaked 2 hours, after the sonicated 30 minutes, put, supply the weight that subtracts mistake with methyl alcohol to room temperature, shake up, filter, filtrate volatilizes, the residue dissolve with methanol, put the 5mL volumetric flask, add methyl alcohol to scale.Shake up, use the pin filtering with microporous membrane, promptly get need testing solution, standby.Parallel 2 parts.
2.4 measure: accurate need testing solution and each 10 μ L injection liquid chromatograph of reference substance solution drawn according to high effective liquid chromatography for measuring, obtains finger-print, as shown in Figure 1.
Embodiment 2. detects the finger-print that 10 batches of summer Sang Ju Yins contain sugared granule
Get 10 batches of mulberry and chrysanthemum granules, detect, get the HPLC collection of illustrative plates of 10 batch samples, as shown in Figure 2 by embodiment 1 condition.By the comparison of 10 batches of HPLC collection of illustrative plates, carry out similarity evaluation, determine that its feature has the peak:
There are 20 features to have the peak in the finger-print, the mean value of existing retention time, peak area and account for total peak area and gather with collection of illustrative plates, specific as follows:
No. 1 peak, average retention time RT is 1.57min, and RSD is 2.06%, and peak area is 1542541, and RSD is 23.75%, accounts for 2.46% of total peak area.
No. 2 peaks, average retention time RT is 1.91min, and RSD is 1.08%, and peak area is 1660403, and RSD is 23.47%, accounts for 2.72% of total peak area.
No. 3 peaks, average retention time RT is 3.56min, and RSD is 1.43%, and peak area is 1572609, and RSD is 24.14%, accounts for 2.84% of total peak area.
No. 4 peaks, average retention time RT is 10.83min, and RSD is 0.55%, and peak area is 3730363, and RSD is 23.20%, accounts for 6.54% of total peak area.
No. 5 peaks, average retention time RT is 11.22min, and RSD is 0.63%, and peak area is 1023133, and RSD is 18.56%, accounts for 1.93% of total peak area.
No. 6 peaks, average retention time RT is 14.60min, and RSD is 0.48%, and peak area is 763899, and RSD is 33.34%, accounts for 1.46% of total peak area.
No. 7 peaks, average retention time RT is 15.25min, and RSD is 0.66%, and peak area is 2077260, and RSD is 24.81%, accounts for 4.52% of total peak area.
No. 8 peaks are chlorogenic acid, and average retention time RT is 19.85min, and RSD is %0.35, and peak area is 2613097, and RSD16.46 is %, accounts for 5.10% of total peak area.
No. 9 peaks, average retention time RT is 20.69min, and RSD is 0.78%, and peak area is 805465, and RSD is 18.00%, accounts for 1.86% of total peak area.
No. 10 peaks, average retention time RT is 22.66min, and RSD is 0.51%, and peak area is 2258797, and RSD is 13.57%, accounts for 4.24% of total peak area.
No. 11 peaks, average retention time RT is 35.66min, and RSD is %1.02, and peak area is 588056, and RSD is 28.63%, accounts for 1.10% of total peak area.
No. 12 peaks, average retention time RT is 36.58min, and RSD is 0.64%, and peak area is 1739577, and RSD is 23.95%, accounts for 3.08% of total peak area.
No. 13 peaks, average retention time RT is 37.48min, and RSD is 0.87%, and peak area is 1144232, and RSD is 29.03%, accounts for 2.28% of total peak area.
No. 14 peaks, average retention time RT is 37.99min, and RSD is 0.61%, and peak area is 1210576, and RSD is 24.05%, accounts for 2.18% of total peak area.
No. 15 peaks, average retention time RT is 39.32min, and RSD is 0.72%, and peak area is 838066, and RSD is 20.36%, accounts for 1.63% of total peak area.
No. 16 peaks, average retention time RT is 40.45min, and RSD is 0.85%, and peak area is 18321566, and RSD is 17.22%, accounts for 31.48% of total peak area.
No. 17 peaks, average retention time RT is 44.58min, and RSD is 0.91%, and peak area is 1925024, and RSD is 20.23%, accounts for 3.40% of total peak area.
No. 18 peaks, average retention time RT is 45.45min, and RSD is 1.13%, and peak area is 1666840, and RSD is 31.43%, accounts for 3.23% of total peak area.
No. 19 peaks, average retention time RT is 48.01min, and RSD is 0.88%, and peak area is 550287, and RSD is 22.56%, accounts for 1.12% of total peak area.
No. 20 peaks, average retention time RT is 55.14min, RSD is 0.77%.Peak area is 3131146, and RSD is 27.56%, accounts for 5.82% of total peak area.
RT is that the peak that 60min occurs later on is a solvent peak.
In the described finger-print, contain the total peak of 20 features in the finger-print of mulberry and chrysanthemum granules, be chlorogenic acid (No. 8 peaks) wherein with reference to the peak, it is the S chromatographic peak, highest peak is 16 chromatographic peaks, the collection of illustrative plates total length is 65min, and the peak of the super total peak area 1% of wherein unimodal area has 20, and they are 1～No. 20 peaks; The peak of the super total peak area 5% of wherein unimodal area has 4, and they are No. 4 peaks, No. 8 peaks (chlorogenic acid), No. 16 peaks, No. 20 peaks; The peak of the super total peak area 10% of unimodal area has 1, is No. 16 peaks.
Similarity evaluation: the chromatographic fingerprints of Chinese materia medica similarity evaluation system software (2004A version) that adopts the Chinese Pharmacopoeia council to recommend carries out comprehensive evaluation to the HPLC collection of illustrative plates, and similarity evaluation the results are shown in Table 2, and the collection of illustrative plates after coupling is seen accompanying drawing 3.
Table 2 HPLC finger-print similarity evaluation
Similarity (contrast) 0.987 0.990 0.991 0.993 0.993 0.993 0.995 0.992 0.994 0.993
Similarity (reference) 0.983 0.974 0.976 0.989 0.987 0.990 0.992 0.979 0.989 1.000
Test sample finger-print and total mode chart are composed the auxiliary similarity evaluation software calculating of machine as calculated, and similarity is greater than 0.90.
The finger-print of the Xia sang ju formulation HPLC common pattern of setting up by said method as shown in Figure 4.
Embodiment 3 summer Sang Ju Yin contains the assay method of sugared granule finger-print
With reference to solution: with embodiment 1
(b) preparation of need testing solution: precision takes by weighing mulberry and chrysanthemum granules 2.0g and puts in the tool plug triangular flask, accurate methyl alcohol 20ml, the jam-pack of adding, soaked 2 hours, after the sonicated 30 minutes, put, supply the weight that subtracts mistake with methyl alcohol to room temperature, shake up, filter, filtrate volatilizes, the residue dissolve with methanol, put the 2mL volumetric flask, add methyl alcohol to scale.Shake up, use the pin filtering with microporous membrane, promptly get need testing solution.
(c) chromatographic condition: chromatographic column is filler (Sunfrie C with the octadecylsilane chemically bonded silica
18, analytical column 150mm * 4.6mm, 5um); Adopting gradient elution, moving phase is 1%L sodium-acetate buffer (A phase) and the gradient eluent (B mutually) of methyl alcohol composition; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 100% 1% acetum, Mobile phase B are 0% methanol solution; In the time of 5 minutes, mobile phase A is that 93% 1% acetum, Mobile phase B are 7% methanol solution; In the time of 9 minutes, mobile phase A is that 90% 1% acetum, Mobile phase B are 10% methanol solution; In the time of 13 minutes, mobile phase A is that 82% 1% acetum, Mobile phase B are 18% methanol solution; In the time of 20 minutes, mobile phase A is that 78% 1% acetum, Mobile phase B are 22% methanol solution; In the time of 35 minutes, mobile phase A is that 60% 1% acetum, Mobile phase B are 40% methanol solution; In the time of 55 minutes, mobile phase A is that 30% 1% acetum, Mobile phase B are 70% methanol solution; In the time of 60 minutes, mobile phase A is that 0% 1% acetum, Mobile phase B are 100% methanol solution; In the time of 65 minutes, mobile phase A is that 100% 1% acetum, Mobile phase B are 0% methanol solution.Column temperature: 30 ℃; Ultraviolet detection: wavelength is 290nm; Flow velocity 1.0mL.min
-1Analysis time: 65min.
(c) measure: the accurate need testing solution 10 μ L that draw inject high performance liquid chromatograph, and according to high effective liquid chromatography for measuring, it is similar to Fig. 1 to obtain finger-print.
Embodiment 4. summer Sang Ju Yin contains the assay method of sugared granule finger-print
(b) preparation of need testing solution: precision takes by weighing mulberry and chrysanthemum granules 10g and puts in the tool plug triangular flask, accurate methyl alcohol 100ml, the jam-pack of adding, soaked 2 hours, after the sonicated 30 minutes, put, supply the weight that subtracts mistake with methyl alcohol to room temperature, shake up, filter, filtrate volatilizes, the residue dissolve with methanol, put the 10mL volumetric flask, add methyl alcohol to scale.Shake up, use the pin filtering with microporous membrane, promptly get need testing solution.
Other is with embodiment 3 operations, and the gained finger-print is similar to Fig. 1.
The assay method of embodiment 5. summer Sang Ju Yin beverage finger-print
The preparation of need testing solution: precision is measured summer Sang Ju Yin beverage 25mL, volatilizes in the water-bath, and the residue dissolve with methanol is put the 5mL volumetric flask, adds methyl alcohol to scale.Shake up, use the pin filtering with microporous membrane, promptly get need testing solution.
Other is with embodiment 3 operations, and the gained finger-print is similar to Fig. 1.
The assay method of embodiment 6. summer mulberry-chrysanthemum oral liquid finger-print
The preparation of need testing solution: precision is measured summer mulberry-chrysanthemum oral liquid 25mL, volatilizes in the water-bath, and the residue dissolve with methanol is put the 5mL volumetric flask, adds methyl alcohol to scale.Shake up, use the pin filtering with microporous membrane, promptly get need testing solution.
Other is with embodiment 3 operating operations, and the gained finger-print is similar to Fig. 1.
Claims (5)
1, a kind of method for building up of Xia sang ju formulation finger-print is characterized in that adopting high performance liquid chromatography, specifically comprises the steps:
(a) preparation of reference substance solution: precision takes by weighing the chlorogenic acid reference substance, is mixed with the solution that every 1ml contains chlorogenic acid 0.2～0.5mg with methyl alcohol, in contrast product solution;
(b) preparation of need testing solution: the A. precision takes by weighing that mulberry and chrysanthemum granules agent powder is dissolved in the methyl alcohol, after the sonicated, filter, and filtrate volatilizes, and the residue dissolve with methanol is put volumetric flask, and molten calmly, membrane filtration promptly gets need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid, volatilizes, and the residue dissolve with methanol is put volumetric flask, and fixed molten, membrane filtration promptly gets need testing solution;
(c) chromatographic condition: chromatographic column is filler with the octadecylsilane chemically bonded silica; Adopt gradient elution, the gradient eluent that moving phase is made up of mobile phase A and Mobile phase B, mobile phase A is that volumetric concentration is 0.1～3% acetum, Mobile phase B is a methyl alcohol; Column temperature: 25～30 ℃; Ultraviolet detection: wavelength is 285～295nm; Flow velocity: 0.5～1.5mLmin
-1Time: 30～80min;
(d) measure: the accurate need testing solution of drawing injects high performance liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
2, the method for building up of Xia sang ju formulation finger-print as claimed in claim 1, its special card is: described step:
(b) preparation of need testing solution: the A. precision takes by weighing mulberry and chrysanthemum granules agent powder 0.1～20g and puts in 20～100mL ground Erlenmeyer flask, accurate methyl alcohol 20～100ml, the jam-pack of adding, after immersion, the sonicated, supply the weight that subtracts mistake, shake up with methyl alcohol, filter, filtrate volatilizes, and the residue dissolve with methanol is put 1～5mL volumetric flask, add methyl alcohol to scale, shake up, membrane filtration promptly gets need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid 25～50mL, volatilizes in the water-bath, and the residue dissolve with methanol is put 1～5mL volumetric flask, adds methyl alcohol to scale, shakes up, and membrane filtration promptly gets need testing solution;
(d) measure: accurate need testing solution 5～25 μ L that draw inject high performance liquid chromatograph, according to high effective liquid chromatography for measuring, obtain finger-print.
3, the method for building up of Xia sang ju formulation finger-print as claimed in claim 1, its special card is: the preparation of described step (b) need testing solution: the A. precision takes by weighing mulberry and chrysanthemum granules agent powder 5.0g and puts in the 50mL ground Erlenmeyer flask, the accurate methyl alcohol 50ml that adds, jam-pack, soaked 2 hours, sonicated was put to room temperature after 30 minutes, supply the weight that subtracts mistake with methyl alcohol, shake up, filter, filtrate volatilizes, the residue dissolve with methanol, put the 5mL volumetric flask, add methyl alcohol, shake up to scale, use membrane filtration, promptly get need testing solution; Or the B. precision measures summer Sang Ju Yin beverage or oral liquid 25mL, volatilizes in the water-bath, and the residue dissolve with methanol is put the 5mL volumetric flask, adds methyl alcohol to scale, shakes up, and membrane filtration promptly gets need testing solution; (c) moving phase is the gradient eluent that 1% acetic acid and methyl alcohol are formed in the chromatographic condition, column temperature: 30 ℃; The detection wavelength is 290nm; Flow velocity 1.0mL.min
-1Analysis time: 65min.
4, the method for building up of Xia sang ju formulation finger-print as claimed in claim 3, its special card is: the gradient elution described in step (c) chromatographic condition, the gradient elution program is carried out with following volumetric concentration configuration:
In the time of 0 minute, mobile phase A is 100%, Mobile phase B is 0%;
In the time of 5 minutes, mobile phase A is 93%, Mobile phase B is 7%;
In the time of 9 minutes, mobile phase A is 90%, Mobile phase B is 10%;
In the time of 13 minutes, mobile phase A is 82%, Mobile phase B is 18%;
In the time of 20 minutes, mobile phase A is 78%, Mobile phase B is 22%;
In the time of 35 minutes, mobile phase A is 60%, Mobile phase B is 40%;
In the time of 55 minutes, mobile phase A is 30%, Mobile phase B is 70%;
In the time of 60 minutes, mobile phase A is 0%, Mobile phase B is 100%;
In the time of 65 minutes, mobile phase A is 100%, Mobile phase B is 0%.
5, the method for building up of Xia sang ju formulation finger-print as claimed in claim 1, it is characterized in that measuring in the finger-print of gained and have 20 features to have the peak, wherein No. 8 chromatographic peaks be chlorogenic acid with reference to the peak, highest peak is 16 chromatographic peaks, the collection of illustrative plates total length is 65min, and is specific as follows:
No. 1 peak, average retention time RT is 1.57min, and RSD is 2.06%, and peak area is 1542541, and RSD is 23.75%;
No. 2 peaks, average retention time RT is 1.91min, and RSD is 1.08%, and peak area is 1660403, and RSD is 23.47%;
No. 3 peaks, average retention time RT is 3.56min, and RSD is 1.43%, and peak area is 1572609, and RSD is 24.14%;
No. 4 peaks, average retention time RT is 10.83min, and RSD is 0.55%, and peak area is 3730363, and RSD is 23.20%;
No. 5 peaks, average retention time RT is 11.22min, and RSD is 0.63%, and peak area is 1023133, and RSD is 18.56%;
No. 6 peaks, average retention time RT is 14.60min, and RSD is 0.48%, and the honeybee area is 763899, and RSD is 33.34%;
No. 7 peaks, average retention time RT is 15.25min, and RSD is 0.66%, and peak area is 2077260, and RSD is 24.81%;
No. 8 peaks are chlorogenic acid, and average retention time RT is 19.85min, and RSD is %0.35, and peak area is 2613097, and RSD is 16.46%;
No. 9 peaks, average retention time RT is 20.69min, and RSD is 0.78%, and peak area is 805465, and RSD is 18.00%;
No. 10 peaks, average retention time RT is 22.66min, and RSD is 0.51%, and peak area is 2258797, and RSD is 13.57%;
No. 11 peaks, average retention time RT is 35.66min, and RSD is %1.02, and peak area is 588056, and RSD is 28.63%;
No. 12 peaks, average retention time RT is 36.58min, and RSD is 0.64%, and peak area is 1739577, and RSD is 23.95%;
No. 13 peaks, average retention time RT is 37.48min, and RSD is 0.87%, and peak area is 1144232, and RSD is 29.03%;
No. 14 peaks, average retention time RT is 37.99min, and RSD is 0.61%, and peak area is 1210576, and RSD is 24.05%;
No. 15 peaks, average retention time RT is 39.32min, and RSD is 0.72%, and peak area is 838066, and RSD is 20.36%.
No. 16 peaks, average retention time RT is 40.45min, and RSD is 0.85%, and peak area is 18321566, and RSD is 17.22%;
No. 17 peaks, average retention time RT is 44.58min, and RSD is 0.91%, and peak area is 1925024, and RSD is 20.23%;
No. 18 peaks, average retention time RT is 45.45min, and RSD is 1.13%, and peak area is 1666840, and RSD is 31.43%;
No. 19 peaks, average retention time RT is 48.01min, and RSD is 0.88%, and peak area is 550287, and RSD is 22.56%;
No. 20 peaks, average retention time RT is 5514min, RSD is 0.77%.Peak area is 3131146, and RSD is 27.56%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100263469A CN100573138C (en) | 2007-01-17 | 2007-01-17 | The foundation of Xia sang ju formulation finger-print and finger-print |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100263469A CN100573138C (en) | 2007-01-17 | 2007-01-17 | The foundation of Xia sang ju formulation finger-print and finger-print |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101034085A CN101034085A (en) | 2007-09-12 |
| CN100573138C true CN100573138C (en) | 2009-12-23 |
Family
ID=38730735
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2007100263469A Active CN100573138C (en) | 2007-01-17 | 2007-01-17 | The foundation of Xia sang ju formulation finger-print and finger-print |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100573138C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101251518B (en) * | 2008-01-31 | 2012-05-30 | 恩施土家族苗族自治州农业科学院 | Method for establishing konjak fingerprint pattern |
| CN102928523B (en) * | 2011-08-11 | 2014-07-30 | 上海雷允上药业有限公司 | Wild chrysanthemum flower fingerprint determination method, its application, and wild chrysanthemum flower quality detection method |
| CN103926344B (en) * | 2014-04-18 | 2015-12-30 | 广州白云山星群(药业)股份有限公司 | The detection method of selfheal finger-print |
| CN105675739B (en) * | 2015-12-31 | 2017-10-31 | 吉林修正药业新药开发有限公司 | A kind of construction method for the HPLC characteristic spectrums for treating anemopyretic cold Chinese medicine |
| CN105717241A (en) * | 2016-04-27 | 2016-06-29 | 广西壮族自治区梧州食品药品检验所 | Determination method of chlorogenic acid in wild chrysanthemum flower |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004117200A (en) * | 2002-09-26 | 2004-04-15 | Kanto Chem Co Inc | Method for analyzing amino acid by high-speed liquid chromatography |
| CN1865988A (en) * | 2006-06-21 | 2006-11-22 | 广州星群（药业）股份有限公司 | Method for detecting amino acid component in Xiasangju preparation |
-
2007
- 2007-01-17 CN CNB2007100263469A patent/CN100573138C/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004117200A (en) * | 2002-09-26 | 2004-04-15 | Kanto Chem Co Inc | Method for analyzing amino acid by high-speed liquid chromatography |
| CN1865988A (en) * | 2006-06-21 | 2006-11-22 | 广州星群（药业）股份有限公司 | Method for detecting amino acid component in Xiasangju preparation |
Non-Patent Citations (6)
| Title |
|---|
| HPLC 法测定桑菊感冒颗粒中绿原酸的含量,. 唐春华，邹晓华.江西中医学院学报，第16卷第1期. 2004 |
| HPLC 法测定桑菊感冒颗粒中绿原酸的含量,. 唐春华，邹晓华.江西中医学院学报，第16卷第1期. 2004 * |
| 反相高效液相色谱法测定夏桑菊颗粒中绿原酸的含量. 王启砚，谢，帆.荆门职业技术学院学报，第20卷第3期. 2005 |
| 反相高效液相色谱法测定夏桑菊颗粒中绿原酸的含量. 王启砚，谢，帆.荆门职业技术学院学报，第20卷第3期. 2005 * |
| 高效液相色谱法测定夏桑菊颗粒中绿原酸的含量. 熊国营，徐君，梁兆昌.江西中医学院学报，第16卷第6期. 2004 |
| 高效液相色谱法测定夏桑菊颗粒中绿原酸的含量. 熊国营，徐君，梁兆昌.江西中医学院学报，第16卷第6期. 2004 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101034085A (en) | 2007-09-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101912440B (en) | Quality control method of complex salvia tablet and application thereof | |
| CN100573138C (en) | The foundation of Xia sang ju formulation finger-print and finger-print | |
| CN101961430B (en) | Quality analysis method of compound Ganmaoling tablets | |
| CN103926345B (en) | The detection method of Xia sang ju formulation characteristic spectrum | |
| CN102579612B (en) | Method for extracting total alkaloid of aconitum soongaricum | |
| CN101315355B (en) | Fingerprint pattern quality determination method for novel healing formulation | |
| CN101320027B (en) | Fingerprint pattern quality determination method of american cockroaches medicinal materials and their extract | |
| CN100362346C (en) | Method for detecting amino acid component in Xiasangju preparation | |
| CN102253135A (en) | Method for establishing fingerprint of Deng's herbal tea granules and standard fingerprint thereof | |
| CN102507832A (en) | Method for measuring high performance liquid chromatography characteristic fingerprint of isatis root and isatis root preparation | |
| CN105911192B (en) | A kind of extracting method and fingerprint atlas detection method of Pterospermi Heterophylli active site promoting blood circulation and removing blood stasis | |
| CN105510488B (en) | A kind of finger-print and its quality determining method for relieving pain patch | |
| CN102890124A (en) | Fingerprint constructing method of total flavonoid components and total alkaloids components in loranthus parasiticus-kudzuvine root preparation and quality detecting method | |
| CN103645251A (en) | Fingerprint spectrum detection method of compound donkey-hide gelatin preparation | |
| CN101672834A (en) | Method for detecting quality of Chinese medicinal preparation for treating diabetic retinopathy | |
| CN103316102B (en) | Detection method for external traditional Chinese medicine preparation for treating haemorrhoids | |
| CN100376894C (en) | Earthworm fingerprint spectrum establishment method and medicinal earthworm identification method | |
| CN102552515A (en) | Quality detection method for blood-nourishing Chinese angelica syrup | |
| CN101744920A (en) | Fingerprint control method for Jian'er Qinjie oral solution | |
| CN104345117A (en) | Qualitative and quantitative detection method of compound Japanese Ardisia Herb tablets | |
| CN100402059C (en) | Quality control method of sinew soothing and pain relieving preparation | |
| CN113189229A (en) | Quality control method of bocepr total alkali | |
| CN104655748A (en) | Erigeron breviscapus granule fingerprint spectrum as well as establishment method and application thereof | |
| CN102269748B (en) | Establishment of mesona blume medicinal material finger print, and finger print thereof | |
| CN110412162B (en) | Construction method of HPLC (high performance liquid chromatography) characteristic spectrum of Baixiangdan capsule and quality detection method of Baixiangdan capsule |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: GUANGZHOU BAIYUNSHAN XINGQUN (PHARMACEUTICAL) CO., Free format text: FORMER NAME: GUANGZHOU XINGQUN (PHARMACEUTICAL) CO., LTD. |
|
| CP03 | Change of name, title or address |
Address after: Haizhuqu District Guangzhou City, Guangdong province 510288 South Island Road No. 162 Patentee after: Guangzhou Xingqun Pharmaceutical Co., Ltd. Address before: Haizhuqu District Prefecture in Guangdong province Guangzhou City Road 510289 No. 162 Patentee before: Guangzhou Xingqun Pharmaceutical Co., Ltd. |