CN106596759A - Taxus chinensis branch and leaf extractive and HPLC analysis method of preparation thereof - Google Patents
Taxus chinensis branch and leaf extractive and HPLC analysis method of preparation thereof Download PDFInfo
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- CN106596759A CN106596759A CN201611105309.2A CN201611105309A CN106596759A CN 106596759 A CN106596759 A CN 106596759A CN 201611105309 A CN201611105309 A CN 201611105309A CN 106596759 A CN106596759 A CN 106596759A
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- ramulus
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- folium taxi
- taxi cuspidatae
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- 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
Abstract
The invention discloses a taxus chinensis branch and leaf extractive and an HPLC analysis method of a preparation thereof. The method comprises the steps that firstly, at least six qualified taxus chinensis branch and leaf extractive samples or other preparation samples are weighed, mixed with methyl alcohol separately and filtered to obtain a sample solution for test of each sample, HPLC analysis is performed, and an HPLC fingerprint chromatography of each sample is obtained, so that the taxus chinensis branch and leaf extractive or an HPLC contrast fingerprint chromatography of the preparation thereof is obtained; then, the samples to be tested and methyl alcohol are mixed and filtered, the sample solutions for test for the samples to be tested are obtained, then, HPLC analysis is performed, and the HPLC fingerprint chromatography of the samples to be tested is obtained; the fingerprint chromatography and the corresponding HPLC contrast fingerprint chromatography are compared, and the advantages and disadvantages of the quality of the taxus chinensis branch and leaf extractive and the preparation thereof are judged on the basis of the comparison result. The quality of the taxus chinensis branch and leaf extractive and the preparation thereof can be well controlled.
Description
Technical field
The invention belongs to pharmaceutical analysiss technical field.More particularly, to a kind of Ramulus et folium taxi cuspidatae extract and its preparation
HPLC analysis methods.
Background technology
Ramulus et folium taxi cuspidatae for taxaceae yew dried leaves, Ramulus et folium taxi cuspidatae mainly taxaneses containing bicyclo-,
Various three rings taxaneses, Colophonium alkanes and Korean pine alkanes diterpene, and lignanoids, flavonoid and phenolic compound etc. into
Point.The effects such as there is Ramulus et folium taxi cuspidatae inducing diuresis to remove edema, the kidney warming to stimulate the menstrual flow, antitumor, suppression diabetes, thus can come as medicine
Utilize.
Because Ramulus et folium taxi cuspidatae has above-mentioned drug effect, thus in the prior art, the extraction of Ramulus et folium taxi cuspidatae obtains many
Side's research.Ramulus et folium taxi cuspidatae extract refers to intermediate extract obtained in the legal techniques of Jing, and its process route is：Take dry Ramulus et folium taxi cuspidatae
Branch and leaf, add water to cook secondary, 2 hours first times, second 1 hour, and collecting decoction, filtration, filtrate is concentrated into relative density
1.12～1.15（60℃）, plus ethanol stands overnight to alcohol content up to 85%, leaching supernatant, 65% ethanol of standby precipitate
Wash twice, merge cleaning mixture, stand overnight, leaching supernatant merges with standby supernatant, reclaim ethanol, add appropriate
Water, mixes, and filters, and filtrate is extremely extracted four times with acetic acid second, merges the cruel liquid of acetic acid second, reclaims acetic acid second extremely and is condensed into thick paste,
It is spray-dried, obtains final product.
Ramulus et folium taxi cuspidatae extract is currently used primarily in and prepares paclitaxel injection, due to without the place of production, Different climate and
Extraction process can all affect Ramulus et folium taxi cuspidatae extract active constituent content, so as to affect the quality of Ramulus et folium taxi cuspidatae extract,
And the quality of Ramulus et folium taxi cuspidatae extract determines the quality of paclitaxel injection, it is therefore desirable to Ramulus et folium taxi cuspidatae
The main component of extract and its preparation is analyzed, and investigates Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations is fine or not and each batch
Mass discrepancy between product.In the prior art, the method that quality analysiss are carried out to different sources Ramulus et folium taxi cuspidatae is only existed,
Specially：Precision weighs 2 g sample powders, adds the ethanol of 20 mL volume fractions 95% to soak the h of supersound process 2 after 24 h under room temperature;
4000rmin-1 is centrifuged 5 min, supernatant liquid filtering, precipitation respectively with 95% ethanol ultrasonic extraction of volume fraction 2 times, every time 30
Min, equal Jing 4000 rmin-1 is centrifuged 5 min, supernatant liquid filtering;Merge 3 filtrates, be placed in Rotary Evaporators and be evaporated;
Then extracted with the mL of V (chloroform): V (water)=1: 1 mixed solvent 10, repeated extraction 3 times, merged three chloromethanes
Alkane phase extract, is evaporated with Rotary Evaporators in 35 DEG C；Extractum methanol dissolves and is settled to 10 mL, with 0.45 μm of micropore
Membrane filtration, filtrate is and supplies test liquid.HPLC analyses, the μ L of sample size 5 are carried out with reference to above-mentioned chromatographic condition.Using following color
Spectral condition carries out eluting：Chinese nation C18 chromatographic columns（4.6 mm × 250 mm, 5 μm）；The nm of Detection wavelength 227；Column temperature is room temperature；
The mLmin-1 of flow velocity 0.8；The μ L of sample size 5.Mobile phase be the aqueous solution of acetonitrile one, binary gradient elutes, 0～20 min, 15%
Acetonitrile is changed to 50%, 20～50 min, and 50% acetonitrile is changed to 100%, 50～60 min, 100% acetonitrile.
In sum, at present for the mass analysis technique of Ramulus et folium taxi cuspidatae extract has the disadvantage that：（1）Mainly
Fingerprint map analyzing is carried out to the Ramulus et folium taxi cuspidatae medical material of different sources, the steady quality of different sources Ramulus et folium taxi cuspidatae is investigated
Property；Impact without considering the factor such as Different climate and extraction process.（2）Contain in finger printing obtained by above-mentioned prior art
There are 12 Characteristic chromatographic peaks, only 3 therein are pointed out, the quantity of information of reflection is little.Therefore, up to the present, still without
Preferably method is come main chemical compositions in analyzing Ramulus et folium taxi cuspidatae extract and its preparation comprehensively and investigates their quality
Stability.
The content of the invention
The technical problem to be solved in the present invention is the defect and technical deficiency for overcoming above-mentioned prior art, there is provided one kind is utilized
A kind of Ramulus et folium taxi cuspidatae extract and its HPLC analysis methods of preparation that high performance liquid chromatography is carried out, carry to Ramulus et folium taxi cuspidatae
Taking the main component of thing and its preparation carries out comprehensive analysis, the fingerprint of Ramulus et folium taxi cuspidatae extract and its preparation to different batches
Collection of illustrative plates is mutually tested, and sets up the HPLC reference fingerprints of Ramulus et folium taxi cuspidatae extract and its preparation, by testing Ramulus et folium taxi cuspidatae
The finger printing of branch and leaf extract and its formulation samples differentiates Ramulus et folium taxi cuspidatae extract with the similarity of reference fingerprint
And its good and bad degree of the quality of the pharmaceutical preparations, reach the purpose of control Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations.
It is an object of the invention to provide the HPLC analysis methods of a kind of Ramulus et folium taxi cuspidatae extract and its preparation.
Above-mentioned purpose of the present invention is achieved through the following technical solutions：
The HPLC analysis methods of a kind of Ramulus et folium taxi cuspidatae extract and its preparation, comprise the steps：
S1. HPLC reference fingerprints are set up
S11. at least 6 qualified Ramulus et folium taxi cuspidatae extract samples or Ramulus et folium taxi cuspidatae extract formulation samples are weighed, respectively
With methanol mixed, filtration, the need testing solution of each sample is obtained；
S12. HPLC analyses are carried out to the need testing solution of each sample of step S11, obtains the HPLC finger printing of each sample, root
The HPLC controls for setting up Ramulus et folium taxi cuspidatae extract or Ramulus et folium taxi cuspidatae extract preparation according to the HPLC finger printing of each sample refer to
Stricture of vagina collection of illustrative plates；
S2. testing sample detection
S21. by testing sample and methanol mixed, filtration, the need testing solution of testing sample is obtained, then carries out HPLC analyses,
Obtain the HPLC finger printing of the testing sample；
S22. the finger printing is compared with corresponding HPLC reference fingerprints, according to comparative result branch of Ramulus et folium taxi cuspidatae is judged
The quality of leaf extract and its quality of the pharmaceutical preparations.
The method can reach the purpose of control Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations.
Wherein it is preferred to, the chromatographic condition of HPLC analyses and mobile phase setting are as follows described in step S12：
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10 min, and concentration is 30% acetonitrile；
Elution time is 10～70 min, and concentration is changed to 45% acetonitrile from 30%；
Elution time is 70～99 min, and concentration is changed to 70% acetonitrile from 45%；
Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；
Elution time is 114～120 min, and concentration is changed to 30% acetonitrile from 90%；
Elution time is 120～130 min, and concentration is 30% acetonitrile.
Preferably, wherein the aqueous solution is pure water.
Preferably, Ramulus et folium taxi cuspidatae extract preparation need to first be crushed into powder described in step S11, make Ramulus et folium taxi cuspidatae extraction
Thing powder formulation sample.
Preferably, before filtering described in step S11, first cool after the mixture ultrasound by sample with methanol.
It is highly preferred that the power of the ultrasound is 40～60kHz.
It is highly preferred that the power of the ultrasound is 60kHz.
It is highly preferred that the time of ultrasound is 20～40min.
It is highly preferred that the time of the ultrasound is 30min.
Preferably, the concentration of step S11 or methanol described in S21 is 100%.
Preferably, filter described in step S11 or S21 is carried out using microporous filter membrane.
It is highly preferred that the aperture of the microporous filter membrane is 0.35～0.45 μm.
It is highly preferred that the aperture of the microporous filter membrane is 0.45 μm.
Preferably, in HPLC analyses described in step S21, equilibration time is 10 min～30 min.
Preferably, in HPLC analyses described in step S21, wavelength is gathered using diode array detector, acquisition range is：
From 190 nm～410 nm of nm～210 nm to 390；The nm of Detection wavelength 227.
In addition, specifically, described in step S12 according to the HPLC finger printing of each sample set up Ramulus et folium taxi cuspidatae extract or
The method of the HPLC reference fingerprints of Ramulus et folium taxi cuspidatae extract preparation is：Install in Chinese Pharmacopoeia Commission on chromatograph
Medicine chromatographic fingerprinting similarity evaluation system, by the system Ramulus et folium taxi cuspidatae extract reference fingerprint, Semen Phaseoli are set up
China fir branch and leaf extract should have 7 common characteristic peaks.
The good and bad standard of Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations is judged described in step S22 according to comparative result
For：If testing sample has 7 characteristic peaks corresponding with reference fingerprint, and similar by chromatographic fingerprints of Chinese materia medica
Degree evaluation system show that it is more than or equal to 0.90 with the similarity of reference fingerprint, then the quality of the testing sample is preferable.
Specifically：The finger printing and reference fingerprint are entered using similarity evaluation
Row compares, and draws the similarity of the finger printing and reference fingerprint；Ramulus et folium taxi cuspidatae is being carried using reference fingerprint
When taking thing sample and being differentiated, in the finger printing of sample, 7 characteristic peaks corresponding with reference fingerprint should be presented, together
When, the finger printing of sample should be not less than 0.90 with the similarity for compareing collection of illustrative plates；If the testing sample have 7 with compare
The corresponding characteristic peak of finger printing, and it is drawn by similarity evaluation and fingerprint image is compareed
The similarity of spectrum is more than or equal to 0.90, it is believed that the quality of the testing sample is preferable.
The present invention draws the HPLC of above-mentioned Ramulus et folium taxi cuspidatae extract and its preparation through substantial amounts of research and exploration, summary
Analysis method, its basic conception is：Set up the HPLC analysis methods of Ramulus et folium taxi cuspidatae extract and its preparation, to different manufacturers,
The Ramulus et folium taxi cuspidatae extract and preparation HPLC fingerprint pattern of different batches carries out comparative study, sets up Ramulus et folium taxi cuspidatae extract
And the HPLC reference fingerprints of preparation, then the finger printing of testing sample is compared, is reflected with the reference fingerprint
Not, fine or not, the quality of Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations are distinguished, control Ramulus et folium taxi cuspidatae extract and its system is reached
The purpose of agent quality.
The invention has the advantages that：
Compared with prior art, the present invention to chromatographic condition due to having carried out reasonable setting so that adopts the method for the present invention
Contain 7 characteristic peaks in the finger printing of gained, and 7 therein are pointed out, compared to gained in prior art
Contain 12 characteristic peaks in finger printing, only 3 therein are pointed out, containing much information for present invention reflection can be preferably
Reach the purpose of control Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations.
Ramulus et folium taxi cuspidatae extract and its HPLC analysis methods of preparation that the present invention sets up, to different manufacturers, difference batch
Secondary Ramulus et folium taxi cuspidatae extract and preparation HPLC fingerprint pattern carries out comparative study, sets up Ramulus et folium taxi cuspidatae extract and preparation
HPLC reference fingerprints, so as to be analyzed to the quality of Ramulus et folium taxi cuspidatae extract and its preparation, Ramulus et folium taxi cuspidatae can be distinguished
Fine or not, the quality of branch and leaf extract and its quality of the pharmaceutical preparations, realizes the purpose of control Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations.
Description of the drawings
Fig. 1 is the reference fingerprint of the Ramulus et folium taxi cuspidatae extract that the embodiment of the present invention 1 is set up.
Fig. 2 is the reference fingerprint of Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae piece that the embodiment of the present invention 2 is set up.
Specific embodiment
The present invention, but embodiment are further illustrated below in conjunction with Figure of description and specific embodiment not to the present invention
Limit in any form.Unless stated otherwise, reagent, the method and apparatus that the present invention is adopted is for the art routinely examination
Agent, method and apparatus.
Unless stated otherwise, agents useful for same of the present invention and material are commercial.
Above-described HPLC is the abbreviation of high performance liquid chromatography.
Embodiment 1
1st, reference fingerprint is set up：
（1）The mg of Ramulus et folium taxi cuspidatae extract 30 is taken, accurately weighed, in putting conical flask with cover, precision adds proper amount of methanol, shakes up,
It is settled to 5 mL, microporous filter membrane（0.45 μm）Filter, obtain final product need testing solution.
（2）Precision draws the μ L sample introductions of need testing solution 10, and using following chromatographic condition eluting is carried out：
Chromatographic column（Phenomenex Luna C18 chromatographic columns（4.6 mm × 250 mm, 5 μm））；
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10min, and concentration is 30% acetonitrile；Elution time is 10
～70 min, concentration is changed to 45% acetonitrile from 30%；Elution time is 70～99 min, and concentration is changed to 70% second from 45%
Nitrile；Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；Elution time be 114～120min, concentration
30% acetonitrile is changed to from 90%；Elution time is 120～130min, and concentration is 30% acetonitrile；Equilibration time 20min；Flow velocity
1 mL·min-1；The μ L of sample size 10；35 DEG C of column temperature；PDA detector acquisition range：200 nm～400nm；Detection wavelength 227
nm。
（3）Determining fingerprint pattern, chromatograph are carried out to the Ramulus et folium taxi cuspidatae extract sample of ten batches by said method
Chinese Pharmacopoeia Commission's similarity evaluation is installed on instrument, Ramulus et folium taxi cuspidatae is set up by the system
Extract reference fingerprint, Ramulus et folium taxi cuspidatae extract should have 7 common characteristic peaks（Wherein there are 7 characteristic peaks to be referred to
Recognize）, as a result as shown in figure 1, wherein：
Peak 1：10-DAB Ⅲ；Peak 2：Baccatin Ⅲ；Peak 3：Taxinine M；Peak 4：10-DAT；Peak 5：
Cephalomannine；Peak 6：7- table -10-DAT；Peak 7：Paclitaxel.
2nd, testing sample detection
（1）The mg of Ramulus et folium taxi cuspidatae extract to be measured 30 is taken, accurately weighed, in putting conical flask with cover, precision adds proper amount of methanol,
Shake up, be settled to 5 mL, microporous filter membrane（0.45 μm）Filter, obtain final product the need testing solution of testing sample.
（2）Precision draws the μ L sample introductions of need testing solution 10, and using following chromatographic condition eluting is carried out：
Chromatographic column（Phenomenex Luna C18 chromatographic columns（4.6mm × 250mm, 5 μm））；
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10 min, and concentration is 30% acetonitrile；Elution time is 10
～70 min, concentration is changed to 45% acetonitrile from 30%；Elution time is 70～99 min, and concentration is changed to 70% second from 45%
Nitrile；Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；Elution time be 114～120 min, concentration
30% acetonitrile is changed to from 90%；Elution time is 120～130 min, and concentration is 30% acetonitrile；The min of equilibration time 20；Stream
1 mLmin-1 of speed；The μ L of sample size 10；35 DEG C of column temperature；PDA detector acquisition range：200 nm～400 nm；Detection wavelength
227 nm。
（3）Obtained after the finger printing of the Ramulus et folium taxi cuspidatae extract sample to be measured, in recycling with above-mentioned method of testing
Medicine chromatographic fingerprinting similarity evaluation system is compared the finger printing and reference fingerprint, draws the finger printing
With the similarity of reference fingerprint.When being differentiated to Ramulus et folium taxi cuspidatae extract sample using reference fingerprint, sample
In the finger printing of product, 7 characteristic peaks corresponding with reference fingerprint should be presented, meanwhile, the finger printing of sample with it is right
0.90 should be not less than according to the similarity of collection of illustrative plates.If the testing sample has 7 features corresponding with reference fingerprint
Peak, and show that it is more than or equal to the similarity of reference fingerprint by similarity evaluation
0.90, it is believed that the quality of the testing sample is preferable.It is red control to be properly arrived at using the reference fingerprint of the present invention
The purpose of bean China fir branch and leaf extract quality.
Embodiment 2
1st, reference fingerprint is set up
（1）Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae piece 6 is taken, is pulverized, weigh the mg of powder 30, in putting conical flask with cover,
Add proper amount of methanol, ultrasonic 30 min to let cool, be settled to 5 mL.Microporous filter membrane（0.45 μm）Filter, obtain final product need testing solution.
（2）Precision draws the μ L sample introductions of need testing solution 10, and using following chromatographic condition eluting is carried out：
Chromatographic column（Phenomenex Luna C18 chromatographic columns（4.6 mm × 250 mm, 5 μm））；
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10 min, and concentration is 30% acetonitrile；Elution time is 10
～70 min, concentration is changed to 45% acetonitrile from 30%；Elution time is 70～99 min, and concentration is changed to 70% second from 45%
Nitrile；Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；Elution time be 114～120 min, concentration
30% acetonitrile is changed to from 90%；Elution time is 120～130 min, and concentration is 30% acetonitrile；The min of equilibration time 20；Stream
1 mLmin-1 of speed；The μ L of sample size 10；35 DEG C of column temperature；PDA detector acquisition range：200 nm～400 nm；Detection wavelength
227 nm。
（3）Finger printing is carried out to the Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae piece of ten batches by said method
It is measured, Chinese Pharmacopoeia Commission's similarity evaluation is installed, by the system on chromatograph
Ramulus et folium taxi cuspidatae piece reference fingerprint is set up, Ramulus et folium taxi cuspidatae piece reference fingerprint there should be 7 common characteristic peaks（Wherein there are 7 spies
Levy peak to be pointed out）, as a result as shown in Fig. 2 wherein：
Peak 1：10-DAB Ⅲ；Peak 2：Baccatin Ⅲ；Peak 3：Taxinine M；Peak 4：10-DAT；Peak 5：
Cephalomannine；Peak 6：7- table -10-DAT；Peak 7：Paclitaxel.
2nd, testing sample detection
（1）Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae piece 6 is taken, is pulverized, weigh the mg of powder 30, in putting conical flask with cover,
Add proper amount of methanol, ultrasonic 30 min to let cool, be settled to 5 mL.Microporous filter membrane（0.45 μm）Filter, obtain final product need testing solution.
（2）Precision draws the μ L sample introductions of need testing solution 10, and using following chromatographic condition eluting is carried out：
Chromatographic column（Phenomenex Luna C18 chromatographic columns（4.6 mm × 250 mm, 5 μm））；
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10 min, and concentration is 30% acetonitrile；Elution time is 10
～70 min, concentration is changed to 45% acetonitrile from 30%；Elution time is 70～99 min, and concentration is changed to 70% second from 45%
Nitrile；Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；Elution time be 114～120 min, concentration
30% acetonitrile is changed to from 90%；Elution time is 120～130 min, and concentration is 30% acetonitrile；The min of equilibration time 20；Stream
1 mLmin-1 of speed；The μ L of sample size 10；35 DEG C of column temperature；PDA detector acquisition range：200 nm～400 nm；Detection wavelength
227 nm。
（3）The fingerprint image of the Ramulus et folium taxi cuspidatae extract preparation to be measured-Ramulus et folium taxi cuspidatae piece sample is obtained with above-mentioned method of testing
Spectrum, recycles similarity evaluation to be compared the finger printing and characteristic spectrum, draws this
Finger printing and the similarity of reference fingerprint, while observing the corresponding characteristic peak number of the finger printing, finally evaluate
The quality of Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae piece sample.Using reference fingerprint to Ramulus et folium taxi cuspidatae extract
When preparation-Ramulus et folium taxi cuspidatae piece sample is differentiated, in the finger printing of sample, should present 7 it is corresponding with reference fingerprint
Characteristic peak, meanwhile, the finger printing of sample should be not less than 0.90 with the similarity of reference fingerprint.If the testing sample
With 7 characteristic peaks corresponding with reference fingerprint, and drawn by similarity evaluation
It is more than or equal to 0.90 with the similarity of reference fingerprint, it is believed that the quality of the testing sample is preferable.Using this
Bright reference fingerprint can be properly arrived at the purpose of control Ramulus et folium taxi cuspidatae extract preparation-Ramulus et folium taxi cuspidatae tablet quality.
Claims (10)
1. HPLC analysis methods of a kind of Ramulus et folium taxi cuspidatae extract and its preparation, it is characterised in that comprise the steps：
S1. HPLC reference fingerprints are set up
S11. at least 6 qualified Ramulus et folium taxi cuspidatae extract samples or Ramulus et folium taxi cuspidatae extract formulation samples are weighed, respectively
With methanol mixed, filtration, the need testing solution of each sample is obtained；
S12. HPLC analyses are carried out to the need testing solution of each sample of step S11, obtains the HPLC finger printing of each sample, root
The HPLC controls for setting up Ramulus et folium taxi cuspidatae extract or Ramulus et folium taxi cuspidatae extract preparation according to the HPLC finger printing of each sample refer to
Stricture of vagina collection of illustrative plates；
S2. testing sample detection
S21. by testing sample and methanol mixed, filtration, the need testing solution of testing sample is obtained, then carries out HPLC analyses,
Obtain the HPLC finger printing of the testing sample；
S22. the finger printing is compared with corresponding HPLC reference fingerprints, according to comparative result branch of Ramulus et folium taxi cuspidatae is judged
The quality of leaf extract and its quality of the pharmaceutical preparations.
2. HPLC analysis methods according to claim 1, it is characterised in that the chromatostrip of HPLC analyses described in step S12
Part and mobile phase setting are as follows：
Acetonitrile-aqueous solution, binary gradient elutes：Elution time is 0～10 min, and concentration is 30% acetonitrile；
Elution time is 10～70 min, and concentration is changed to 45% acetonitrile from 30%；
Elution time is 70～99 min, and concentration is changed to 70% acetonitrile from 45%；
Elution time is 99～114 min, and concentration is changed to 90% acetonitrile from 70%；
Elution time is 114～120 min, and concentration is changed to 30% acetonitrile from 90%；
Elution time is 120～130 min, and concentration is 30% acetonitrile.
3. HPLC analysis methods according to claim 1, it is characterised in that Ramulus et folium taxi cuspidatae extract described in step S11
Preparation need to first be crushed into powder, and make Ramulus et folium taxi cuspidatae extract powder formulation sample.
4. HPLC analysis methods according to claim 1, it is characterised in that before filtering described in step S11, first by sample
Cool after ultrasonic with the mixture of methanol.
5. HPLC analysis methods according to claim 4, it is characterised in that the power of the ultrasound is 40～60kHz, is surpassed
The time of sound is 20～40min.
6. HPLC analysis methods according to claim 1, it is characterised in that filter described in step S11 or S21 and adopt micropore
Filter membrane is carried out.
7. HPLC analysis methods according to claim 6, it is characterised in that the aperture of the microporous filter membrane is 0.35～
0.45μm。
8. HPLC analysis methods according to claim 1, it is characterised in that according to the HPLC of each sample described in step S12
Finger printing sets up the method for the HPLC reference fingerprints of Ramulus et folium taxi cuspidatae extract or Ramulus et folium taxi cuspidatae extract preparation：
Chinese Pharmacopoeia Commission's similarity evaluation is installed on chromatograph, Semen Phaseoli is set up by the system
China fir branch and leaf extract reference fingerprint, Ramulus et folium taxi cuspidatae extract should have 7 common characteristic peaks.
9. HPLC analysis methods according to claim 1, it is characterised in that judge according to comparative result described in step S22
The good and bad standard of Ramulus et folium taxi cuspidatae extract and its quality of the pharmaceutical preparations is：If testing sample has 7 and reference fingerprint
Corresponding characteristic peak, and show that it is similar to reference fingerprint by similarity evaluation
Degree is more than or equal to 0.90, then the quality of the testing sample is preferable.
10. HPLC analysis methods according to claim 1, it is characterised in that the concentration of methanol described in step S11 or S21
For 100%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109324127A (en) * | 2018-09-30 | 2019-02-12 | 东阳市杰迩威生物科技有限公司 | The detection method of taxol in Chinese yew extract |
| CN110441439A (en) * | 2019-07-01 | 2019-11-12 | 杭州师范大学 | It is a kind of for distinguishing the metabolic markers and its detection method of heavy foliage Chinese yew and taxusyunnanensis |
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| CN105388234A (en) * | 2015-12-24 | 2016-03-09 | 湖北祥瑞丰生态果业有限公司 | Detection method for content of paclitaxel in taxus chinensis branch |
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| CN109324127A (en) * | 2018-09-30 | 2019-02-12 | 东阳市杰迩威生物科技有限公司 | The detection method of taxol in Chinese yew extract |
| CN110441439A (en) * | 2019-07-01 | 2019-11-12 | 杭州师范大学 | It is a kind of for distinguishing the metabolic markers and its detection method of heavy foliage Chinese yew and taxusyunnanensis |
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| CN106596759B (en) | 2019-07-02 |
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