CN101708235A - Finger print quality detecting method by pollen pini ethanol extract intermediates for strengthening body resistance and dissolving stasis - Google Patents
Finger print quality detecting method by pollen pini ethanol extract intermediates for strengthening body resistance and dissolving stasis Download PDFInfo
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- CN101708235A CN101708235A CN200910201483A CN200910201483A CN101708235A CN 101708235 A CN101708235 A CN 101708235A CN 200910201483 A CN200910201483 A CN 200910201483A CN 200910201483 A CN200910201483 A CN 200910201483A CN 101708235 A CN101708235 A CN 101708235A
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Abstract
The invention relates to a finger print quality detecting method by pollen pini ethanol extract intermediates for strengthening body resistance and dissolving stasis. The method comprises the following steps of: (1) extracting pollen pini intermediates with ethanol to prepare a sample product; (2) carrying out gradient elution on a mobile phase; (3) establishing a standard finger print: selecting 10 different batches of pollen pini ethanol extract intermediates for strengthening body resistance and dissolving stasis for carrying out high-efficiency liquid chromatogram finger print analysis; comparing all peaks in an HPLC finger print to determine 3 common peaks; sequentially numbering the peaks according to peak time (finger 1); and (4) controlling the quality of the finger print: comparing a sample HPLC finger print with the standard finger print, wherein a similarity obtained by calculating the 3 common peaks cannot be less than 0.8. The detecting method can comprehensively control the quality of the pollen pini ethanol extract intermediates for strengthening body resistance and dissolving stasis so as to guarantee the stable quality of the products.
Description
Technical field
The invention belongs to Pollen Pini alcohol extraction intermediate quality testing field, particularly relate to a kind of supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate finger print quality detecting method.
Background technology
Medicine compositions for strengthening healthy qi to remove blood stasis is made up of Radix Salviae Miltiorrhizae, fermented Cordyceps powder, Semen Persicae, Pollen Pini, Herb Gynostemmae Pentaphylli, Fructus Schisandrae Chinensis Six-element medicine.Go up two kinds of dosage forms of municipalization at present: (1) FUZHENG HUAYU JIAONANG agent: obtained China national New Drug Certificate (traditional Chinese medicines card word Z20020053) in 2002, and approval national drug standards WS
3-459 (Z-79)-2005 (Z); (2) supporting vital QI and dispersing blood stasis tablet: obtained national New Drug Certificate (traditional Chinese medicines card word Z20050564) in 2005, and approval national drug standards YBZ19332005-2009Z.Supporting vital QI and dispersing blood stasis has effects such as soothing liver-QI is invigorated blood circulation, beneficial intensive culture liver, dissolving blood stasis and detoxication, hard masses softening and resolving, enhancing human body immunity function, cures mainly chronic hepatitis Bhepatic fibrosis, early stage liver cirrhosis.
The effective ingredient nature difference of considering supporting vital QI and dispersing blood stasis preparation Six-element medical material is bigger, therefore selects different extracting method for use at different flavour of a drug in process of production.Wherein Pollen Pini adopts warm macerating technology, and is specific as follows: with Pollen Pini under 40 ℃ of conditions, with 50% ethanol warm macerating 2 times, 4h for the first time, 2h for the second time, the merging leachate is concentrated into relative density 1.1-1.2 (50 ℃-55 ℃), promptly gets Pollen Pini alcohol extraction intermediate.
In recent years, fingerprint pattern technology has become the important means of Chinese medicine compound quality control and evaluation, consults the document of delivering both at home and abroad, does not up to the present have the report of supporting vital QI and dispersing blood stasis preparation Pollen Pini alcohol extraction intermediate fingerprint map analyzing.Therefore the present invention will remedy the blank of supporting vital QI and dispersing blood stasis preparation Pollen Pini alcohol extraction intermediate finger printing research.
Summary of the invention
Technical problem to be solved by this invention provides a kind of supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate finger print quality detecting method, and this method can be controlled the quality of supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate comprehensively, to guarantee the steady quality of product.
A kind of supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate finger print quality detecting method of the present invention comprises:
(1) Pollen Pini alcohol extraction intermediate test sample preparation
Take by weighing Pollen Pini alcohol extraction intermediate 1.00 grams, put in the 10mL volumetric flask, add 50% methanol, 5～8ml, supersound process 20min adds 50% methanol to scale, shakes up, and crosses the 0.45um microporous filter membrane, gets the need testing solution of subsequent filtrate as Pollen Pini alcohol extraction intermediate;
(2) gradient elution mobile phase
With octadecylsilane chemically bonded silica is filler, and condition of gradient elution is:
| Time (min) | A phase: 0.05% phosphoric acid-water (%) | B phase: acetonitrile (%) |
| ??0 | ??95 | ??5 |
| Time (min) | A phase: 0.05% phosphoric acid-water (%) | B phase: acetonitrile (%) |
| ??60 | ??60 | ??40 |
| ??120 | ??10 | ??90 |
Chromatographic column C18 post, mobile phase A are 0.05% phosphoric acid-water mutually, and B is acetonitrile mutually; Gradient elution: A phase 0-60-120min, 95-60-10%, B phase 5-40-90%, flow velocity 0.8-1.2ml/min detects wavelength 254nm, and column temperature 20-40 ℃, sample size 10-20ul;
(3) foundation of standard finger-print
Choose the medicine compositions for strengthening healthy qi to remove blood stasis Pollen Pini alcohol extraction intermediate of 10 batches of different batches, carry out the efficient liquid-phase chromatograph finger print atlas analysis according to above-mentioned technology contents.All peaks in the HPLC finger printing are compared, determined 3 total peaks, and be numbered (Fig. 1) by appearance time precedence;
(4) quality control of finger printing
Adopt Chinese medicine fingerprint area of computer aided similarity evaluation software (Chinese Pharmacopoeia Commission), the HPLC finger printing and the standard finger-print (or reference substance common pattern finger printing) of sample compared.Should be with the similarity that 3 total peaks calculate less than 0.8.
Beneficial effect
1, uses stratographic fingerprint characteristic,, more effectively control the quality of product intermediate by the quantization parameter that finger printing obtains;
2, investigated the stability of need testing solution, the result shows, at room temperature, within 24 hours, it is stable that medicine compositions for strengthening healthy qi to remove blood stasis Pollen Pini alcohol extraction intermediate need testing solution keeps, and precision and replica test show that all similarity is more than 0.9;
3, through the finger printing similarity of 10 batches of supporting vital QI and dispersing blood stasis preparation Pollen Pini alcohol extraction intermediate is calculated, the similarity of finding Pollen Pini alcohol extraction intermediate is between 0.8-1.0.The supporting vital QI and dispersing blood stasis preparation Pollen Pini alcohol extraction intermediate finger printing quality control method of setting up is described, good reproducibility, specificity is strong, easy and simple to handle, method is reliable, can carry out science, estimate effectively supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate quality, and confirm to apply to quality control in the actual production, satisfy the demand of enterprise control of product quality.
4, provide condition for the supporting vital QI and dispersing blood stasis preparation carries out the finger printing quality control, also guaranteed the steady quality of supporting vital QI and dispersing blood stasis preparation simultaneously.
Description of drawings
Fig. 1 is a supporting vital QI and dispersing blood stasis preparation Pollen Pini alcohol extraction intermediate standard HPLC finger printing;
Fig. 2 is a supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate sample HPLC fingerprint image spectrogram.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
The preparation of finished product, reagent and instrument:
Finished product: supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate source: Shanghai Huang Hai Pharmaceutical Co. Ltd
Extraction process: with Pollen Pini under 40 ℃ of conditions, with 50% ethanol warm macerating 2 times, 4h for the first time, 2h merges leachate for the second time, is concentrated into relative density 1.1-1.2 (50 ℃-55 ℃), promptly gets Pollen Pini alcohol extraction intermediate;
Reagent: methanol (analytical pure) source: Shanghai development chemical industry one factory;
Phosphoric acid (analytical pure) source: Shanghai amalgamation factory;
Acetonitrile (chromatographically pure) source: Tedia company Inc;
Water is pure water.
Instrument:
| The instrument title | Model | Production unit |
| Electronic balance | The AL104 type | ??METTLER?TOLEDO |
| High performance liquid chromatograph | ??HP1100 | ??Agilent |
| The chromatographic column calorstat | The HT-130 type | Tianjin Hengao Technology Development Co., Ltd. |
| The water system microporous filter membrane | ??0.45um | Homemade |
| Ultrasonic cleaner | ??KQ-50E | Kunshan ultrasonic instrument company limited |
Take by weighing Pollen Pini alcohol extraction intermediate 1.00 grams, put in the 10mL volumetric flask, add 50% methanol, 5～8ml, supersound process 20min adds 50% methanol to scale, shakes up, cross the 0.45um microporous filter membrane, get the need testing solution of subsequent filtrate as supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate;
Chromatographic condition:
Column temperature: 30 ℃, detect wavelength: 254nm, sample size: 10 μ l, chromatographic column: Alltima (250mm * 4.6mm, 5 μ m).Elution requirement sees Table 1,
Table 1 elution requirement
| Time (min) | A phase: 0.05% phosphoric acid-water (%) | B phase: acetonitrile (%) |
| ??0 | ??95 | ??5 |
| ??60 | ??60 | ??40 |
| ??120 | ??10 | ??90 |
After the determining fingerprint pattern condition is determined, by at least three batch samples are measured, calculate relevant parameter, and propose the suitable parameters condition of highly effective liquid phase chromatographic system on this basis.Promptly No. 3 chromatographic peak area repeatability (RSD) are not higher than 3%, and retention time repeatability (RSD) is not higher than 2%; The chromatographic column post is imitated and is no less than 15,000 theoretical cam curves in No. 3 chromatographic peaks.
Medicine compositions for strengthening healthy qi to remove blood stasis Pollen Pini alcohol extraction intermediate sample fingerprint image spectrogram is seen Fig. 2.
Supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate sample to 10 batches carries out fingerprint map analyzing, adopts Chinese medicine fingerprint area of computer aided similarity evaluation software (2004A version, Chinese Pharmacopoeia committee) that the similarity of finger printing is calculated.Adopt total peak pattern, the setting-up time window width is 0.1min, generate reference fingerprint with the average method, the HPLC finger printing and the standard finger-print of sample are compared, the result shows that the finger printing of 10 batches of supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate samples and the similarity of standard finger-print are all between 0.8-1.0.The supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate finger printing quality control method of setting up is described, easy and simple to handle, method is reliable, and confirmation can apply to quality control in the actual production, satisfies the demand of enterprise to control of product quality.
Claims (1)
1. supporting vital QI and dispersing blood stasis Pollen Pini alcohol extraction intermediate finger print quality detecting method comprises:
(1) Pollen Pini alcohol extraction intermediate test sample preparation
Take by weighing Pollen Pini alcohol extraction intermediate 1.00 grams, put in the 10mL volumetric flask, add 50% methanol, 5～8ml, supersound process 20min adds 50% methanol to scale, shakes up, and crosses the 0.45um microporous filter membrane, gets the need testing solution of subsequent filtrate as Pollen Pini alcohol extraction intermediate;
(2) gradient elution mobile phase
With octadecylsilane chemically bonded silica is filler, and condition of gradient elution is: mobile phase A is 0.05% phosphoric acid-water mutually, and B is acetonitrile mutually; Gradient elution: A phase 0-60-120min, 95-60-10%, B phase 5-40-90%, flow velocity 0.8-1.2ml/min detects wavelength 254nm, and column temperature 20-40 ℃, sample size 10-20ul;
(3) foundation of standard finger-print
The medicine compositions for strengthening healthy qi to remove blood stasis Pollen Pini alcohol extraction intermediate of choosing 10 batches of different batches carries out the efficient liquid-phase chromatograph finger print atlas analysis, and all peaks in the HPLC finger printing are compared, and has determined 3 total peaks, and has been numbered by appearance time precedence;
(4) quality control of finger printing
The HPLC finger printing and the standard finger-print of sample are compared, should be with the similarity that 3 total peaks calculate less than 0.8.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110596259A (en) * | 2019-06-20 | 2019-12-20 | 烟台新时代健康产业有限公司 | Method for identifying storage time of pine pollen based on metabonomics analysis |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110596259A (en) * | 2019-06-20 | 2019-12-20 | 烟台新时代健康产业有限公司 | Method for identifying storage time of pine pollen based on metabonomics analysis |
| CN110596259B (en) * | 2019-06-20 | 2022-03-01 | 烟台新时代健康产业有限公司 | Method for identifying storage time of pine pollen based on metabonomics analysis |
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