CN105486764B - A kind of analysis method and its application of the gas-phase fingerprint pattern of female clever ball - Google Patents
A kind of analysis method and its application of the gas-phase fingerprint pattern of female clever ball Download PDFInfo
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- 238000000034 method Methods 0.000 claims abstract description 60
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- Treatment Of Liquids With Adsorbents In General (AREA)
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of gas-phase fingerprint pattern analysis methods of female clever ball, which comprises the foundation of female spirit ball standard gas-phase fingerprint pattern, the measurement of female clever ball gas-phase fingerprint pattern to be measured and similarity system design judgement and etc..The present invention has finally determined the chromatographic condition of the gas-phase fingerprint pattern analysis of female clever ball, investigation has determined the control gas-phase fingerprint pattern at 10 shared peaks, has the characteristics that method is easy, speed is fast, favorable reproducibility by attempting multiple programs Elevated Temperature Conditions.The standard gas-phase fingerprint pattern established meets related request through methodology validation, and strong applicability can more fully reflect product quality situation, and standard is controlled and formulated for quality and provides important evidence.
Description
Technical field
The present invention relates to a kind of gas-phase fingerprint pattern analysis methods of female clever ball, belong to Pharmaceutical Analysis technical field.
Background technique
Female spirit ball is a kind of compound Chinese medicinal preparation, and major function is menstruation regulating blood nourishing, removing blood stasis and bringing forth new life.For irregular menstruation,
More or less, menalgia, cold uterus fail to be impregnated for a long time, and habitual abortion, leukorrhea with reddish discharge, metrorrhagia and metrostaxis are more than, the long deficiency of vital energy of disease, kidney
Thanks to pain in the loins.
The prescription of female spirit ball is as follows:
[prescription]
(1) north rhizoma cyperi (system) 37g Radix Glycyrrhizae 7g radix cynanchi atrati 14g motherwort 14g Radix Astragali 14g cockscomb 14g Radix Ophiopogonis 14g
Schisandra chinensis 14g glutinous rehmannia 14g safflower 14g caulis akebiae 10g Rhizoma Atractylodis Macrocephalae (stir-fry) 14g red halloysite 14g Poria cocos 14g Cortex Magnoliae Officinalis 10g meat desert
Rong (system) 14g Radix Paeoniae Alba (parched with wine) 14g
(2) rhizoma cyperi (system) 37g schizonepeta 10g cortex moutan 14g donkey-hide gelatin 14g Radix Angelicae Sinensis 14g rhizoma ligustici 10g red ginseng 14g deer horn
Glue 14g bulbus fritillariae cirrhosae 14g myrrh (stir-fry) 14g fructus amomi 14g rhizoma corydalis 14g fennel seeds (salt system) 14g colla carapacis et plastri testudinis 14g Rhizoma Chuanxiong
14g
17 taste of [preparation method] above prescription (one), adds water to cook secondary, 3 hours for the first time, second 2 hours, merges and decocts
Liquid, filtration, filtrate are placed 24 hours, are taken supernatant that rice vinegar 280g is added, are condensed into thick paste.Above (two) ten five tastes of prescription, powder
It is broken into middle powder, is added in above-mentioned thick paste, mixes, is dried, crushed into fine powder, is sieved, mixes, with the general ball of yellow rice wine, 1800 balls is made,
Sugar coating or film-coating, polishing, it is dry to get.
Due to chemical composition of Chinese materia medica complexity, mechanism of action is indefinite, and existing Chinese medicine pharmaceutical technology is caused to be difficult to ensure Chinese medicine
Stable product quality, the type and quantity that can reflect Chinese medicine intrinsic chemical ingredient comprehensively that traditional Chinese medicine fingerprint is established, into
And reflect the quality of Chinese medicine.The middle the effective elements of the medicine overwhelming majority is not clear at this stage, by the way of traditional Chinese medicine fingerprint,
Traditional Chinese medicine quality will effectively be characterized.Finger-print is approved by international community, is conducive to Chinese medicine and product enters international market.
Female spirit ball current standard is recorded in Drug Standard of Ministry of Public Health of the Peoples Republic of China " Traditional Chinese medicine historical preparation " the tenth,
Any qualitative, quantitative item controlled for flavour of a drug is not directed in standard, currently, not carrying out finger-print research to female clever ball
Report, and finger-print can sufficiently characterize the globality and complexity of Chinese materia medica preparation, therefore inventor is to the effective of female clever ball
Ingredient is studied.It was found that wherein containing there are many water-soluble and Alcohol soluble composition, the present inventor targetedly to these at
Divide and studied, establish a kind of standard finger-print of female clever ball, matter is carried out to the female clever ball in production using the map
Amount supervision, to ensure product quality.
Summary of the invention:
The present invention provides a kind of female clever ball gas chromatography standard reference fingerprint, and method for building up is as follows:
1) prepared by test solution:
Female clever ball is taken, adds water, ultrasonic extraction, extracting solution adds ethyl acetate to extract, and extract liquor ethyl acetate layer adds acetic acid second
Ester is settled to scale, shakes up, and crosses 0.22 μm of miillpore filter, both；
2) gas chromatogram is obtained:
Test solution gas chromatograph, obtains gas chromatogram；
3) reference fingerprint is generated
10 shared peaks have been determined in the gas-phase fingerprint pattern of more batches of qualified female clever ball samples of selection, have utilized Chinese medicine chromatography
Fingerprint similarity evaluation system (2.0 editions), fitting determine female clever ball reference fingerprint.
The standard reference material of effective component can be added as needed using as object of reference peak.
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition is shown in Table 1；Injector temperature: 150~200 DEG C；Detector: FID；Temperature: 230~280
℃；Carrier gas: nitrogen；Split sampling；Split ratio: 1: 1~5: 1；Sample volume: 0.5~1 μ l.
1 temperature programming table of table
Temperature programming table is preferred:
Wherein, the method for building up of preferred standard control finger-print is as follows:
1) preparation of test solution, the method is as follows:
It taking female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 0.8~1.2g, adds 20~30mL of water,
It is ultrasonically treated 20~40min, adds ethyl acetate to extract 2 times, the 1st 10~20ml, the 2nd 5~15ml, combining extraction liquid to 15
In~35ml volumetric flask, add ethyl acetate to be settled to scale, shake up, cross 0.22 μm of miillpore filter to get；
It is preferred that taking female clever ball appropriate, dragee removes sugar-coat, is ground into fine powder, and precision weighs 1.0g, adds water 25mL, ultrasound
30min is handled, adds ethyl acetate to extract 2 times, first time 15ml, second of 10ml, combining extraction liquid shakes into 25ml volumetric flask
It is even, cross 0.22 μm of miillpore filter to get.
2) gas chromatogram is obtained:
Test solution gas chromatograph, obtains gas chromatogram；
Standard reference material solution is wherein prepared using Paeonol as object of reference, the method is as follows:
It takes Paeonol reference substance appropriate, adds ethyl acetate that the solution that every 1mL contains 10~15 μ g is made, it is molten as reference substance
Liquid, it is excellent to take Paeonol reference substance appropriate, add ethyl acetate be made every 1mL containing the solution to about 12.55 μ g of Paeonol to get.
3) reference fingerprint is generated, the method is as follows:
Test solution and control solution are injected into gas chromatograph, obtain chromatogram,
Analysis comparison is carried out to the gas-phase fingerprint pattern of more batches of batch female clever balls, it is determined that 10 shared peaks, in chromatogram, with
Retention time identical chromatography in Paeonol reference substance peak is No. 5 peaks referring to peak, calculates the relative retention time at other 9 shared peaks
It is respectively as follows: 0.365 (1), 0.372 (2), 0.769 (3), 0.772 (4), 1.000 (5), 1.263 (6), 1.283 (7), 1.359
(8), 1.395 (9), 1.565 (10), relative deviation utilize chromatographic fingerprints of Chinese materia medica similarity evaluation within ± 3%
System (2.0 editions), fitting, which determines, obtains female clever ball standard control gas-phase fingerprint pattern.
The present invention also provides a kind of Gas Chromatography Fingerprint detection methods of female clever ball, which is characterized in that including as follows
Step
1) preparation of sample to be tested chromatogram:
Female clever ball is taken, adds water, ultrasonic extraction, extracting solution adds ethyl acetate to extract, and extract liquor ethyl acetate layer adds acetic acid second
Ester is settled to scale, shakes up, and crosses 0.22 μm of miillpore filter, takes filtrate, injects gas chromatograph, obtains chromatogram；
2) sample to be tested chromatogram and standard control fingerprint similarity calculate:
The chromatogram of sample to be tested is imported to " similarity evaluation A editions " of Chinese Pharmacopoeia committee
Software compares the chromatogram and standard control fingerprint similarity of sample to be tested；
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition is shown in Table 1；Injector temperature: 150~200 DEG C；Detector: FID；Temperature: 230~280
℃；Carrier gas: nitrogen；Split sampling；Split ratio: 1: 1~5: 1；Sample volume: 0.5~1 μ l.
1 temperature programming table of table
Temperature programming table is preferred:
Wherein, preferred detection method of the invention is as follows:
1) preparation of sample to be tested chromatogram:
It taking female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 0.8~1.2g, adds 20~30mL of water,
It is ultrasonically treated 20~40min, adds ethyl acetate to extract 2 times, the 1st 10~20ml, the 2nd 5~15ml, combining extraction liquid to 15
In~35ml volumetric flask, add ethyl acetate to be settled to scale, shake up, cross 0.22 μm of miillpore filter, take filtrate, injects gas-chromatography
Instrument obtains chromatogram；
It is preferred that taking female clever ball appropriate, dragee removes sugar-coat, is ground into fine powder, and precision weighs 1.0g, adds water 25mL, ultrasound
30min is handled, adds ethyl acetate to extract 2 times, first time 15ml, second of 10ml, combining extraction liquid shakes into 25ml volumetric flask
It is even, cross 0.22 μm of miillpore filter to get.
2) sample to be tested chromatogram and standard control fingerprint similarity calculate:
The chromatogram of sample to be tested is imported to " similarity evaluation A editions " of Chinese Pharmacopoeia committee
Software compares the chromatogram and standard control fingerprint similarity of sample to be tested；Similarity is that 0.90 the above are qualifications, similar
For degree for 0.95 the above are being consistent, similarity is that 0.98 the above are identical.
The present invention further establishes the analysis method of female clever ball, using the party by the foundation of the above finger-print
Method can be obtained identification conclusion, more connect with standard finger-print with the female clever ball of accurate discrimination according to the comparison with standard finger-print
Closely, product quality is better, and computer approach confirmation can be used in similarity, if similarity is that 0.90 the above are qualification, similarities
For 0.95 the above are being consistent, similarity is that 0.98 the above are identical.
The preparation method that the above method is especially the standard control gas-phase fingerprint pattern of female clever ball is obtained by screening,
The following are screening processes:
1 instrument and reagent
Instrument: U.S. Agilent7890A, Agilent Open LAB Control Panel work station.
Chromatographic column: 1. non-polar column, stationary phase is using 5% phenyl, 95% dimethyl polysiloxane as filler, Agilent HP-
5(30m×0.32mm×0.25μm)；
2. middle polar column, stationary phase is using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, Agilent
DB-1701(30m×0.25mm×0.25μm)。
3. middle polar column, 50% diphenyl of stationary phase, 50% dimethyl polysiloxane, Agilent DB-17 (30m ×
0.25mm×0.15μm)
Reagent: ethyl acetate (chromatographically pure, Tianjin Concord Technology Co., Ltd.)；Water is ultrapure water.
Reference substance: Paeonol (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110708-200506, assay use)；
α-cyperolone (National Institute for Food and Drugs Control, lot number: 110748-201111, assay use)；Honokiol (China
Food and medicine examines and determine research institute, lot number: 110730-201011, assay is used, by 98.4% purity meter)
Sample: female spirit ball (Tianjin Tasly (Liaoning) Pharmaceutical Co., Ltd.), totally 11 batches, specific lot number is shown in Table 3,
It is qualified to examine through Product Quality Verification Centers.
3 sample message table of table
The selection of 2 GC conditions
The selection of 2.1 chromatographic columns
Because female clever ball includes two kinds of specifications of dragee and film-coating ball, and dragee sugar-coat agent weight gain large percentage is (about
40%) using vegetable pill when, therefore method is investigated first, (lot number: 20130209) sample carries out, and is directed to again after condition determines substantially
Dragee and film-coating ball carry out methodological study.
It referring to pertinent literature, investigates opposed polarity capillary column and is tested, as a result 3 kinds of opposed polarity capillary column separation
Obtained chromatographic peak number and peak shape is almost the same, (see Fig. 1~3), but wherein the chromatographic peak peak shape of chromatographic column 2. is preferable, therefore makes
The investigation of temperature programming condition is 2. carried out with chromatographic column.
The selection of 2.2 temperature programming conditions
Investigation adjustment 2. is carried out to temperature programming condition using chromatographic column, in the case where guaranteeing the complete situation of appearance, as far as possible
Shorten analysis duration.As a result it after having adjusted more than 30 secondary program Elevated Temperature Conditions, determines in this temperature programming condition (being shown in Table 2-1)
Under, about 11 chromatographic peaks are isolated, analysis time overall length is 55min, sees Fig. 4.
Table 2-1 temperature programming table
The determination of 3 GC conditions
It to sum up analyzes, finally determines that female clever ball gas-phase fingerprint pattern chromatographic condition is as follows:
Chromatographic column: stationary phase is using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, Agilent DB-
1701(30m×0.25mm×0.25μm).Column temperature: temperature programming condition is shown in Table 1；Injector temperature: 180 DEG C；Detector: FID；
Temperature: 250 DEG C；Carrier gas: nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ L.
1 temperature programming table of table
The selection of 4 test article treating methods
4.1 extract the selection of solvent using dragee first:
1. taking dragee appropriate, it is ground into fine powder, precision weighs 7g (converting with vegetable pill), adds ethyl acetate 25mL, at ultrasound
30min is managed, centrifuging and taking supernatant is concentrated to dryness, and residue adds ethyl acetate to redissolve and is settled to 5ml, is shaken up, and 0.45 μm of micropore is crossed
Filter membrane to get.
2. taking dragee appropriate, it is ground into fine powder, precision weighs 7g (converting with vegetable pill), adds hexamethylene 25mL, be ultrasonically treated
30min, centrifuging and taking supernatant, is concentrated to dryness, and residue adds hexamethylene to redissolve and is settled to 5ml, shakes up, and crosses 0.45 μm of micropore filter
Film to get.
3. taking dragee appropriate, it is ground into fine powder, precision weighs 7g, adds water 25mL, is ultrasonically treated 30min, hexamethylene is added to extract
It taking 2 times, each 20ml, combining extraction liquid is recycled to dry, and residue adds hexamethylene to redissolve, and is settled in 5ml volumetric flask, it shakes up,
Cross 0.45 μm of miillpore filter to get.
4. taking dragee appropriate, it is ground into fine powder, precision weighs 7g, adds water 25mL, is ultrasonically treated 30min, adds ethyl acetate
Extraction 2 times, each 20ml, combining extraction liquid are recycled to dry, and residue adds ethyl acetate to redissolve, and is settled in 5ml volumetric flask, shakes
It is even, cross 0.45 μm of miillpore filter to get.
As a result see Fig. 5~Fig. 8, be 4. that water mentions ethyl acetate and extracts using test article treating method as seen from the figure
The chromatogram information arrived is most, therefore determines using test article treating method 4., continues to investigate.
4.2 according to above-mentioned investigation as a result, extracting the selection of mode using dragee, i.e. 5. test article treating method takes
Appropriate dragee is ground into fine powder, and precision weighs 7g, adds water 25mL, is heated to reflux 30min, adds ethyl acetate extraction 2 times, every time
20ml, combining extraction liquid are recycled to dry, and residue adds ethyl acetate to redissolve, and is settled in 5ml volumetric flask, shake up, and cross 0.45 μm
Miillpore filter to get.
As a result see Fig. 9, as seen from the figure, except retention time is in addition to 3 chromatographic peaks before 10min obviously become larger,
Remaining chromatographic peak obviously becomes smaller, therefore considers that the globality of gas-phase fingerprint pattern information and the simplicity of test sample operation, selection supply
Test product processing method is 4..
4. 4.3 use above-mentioned test article treating method, during carrying out dragee methodological study, gas phase sample introduction needle occur
The phenomenon that bending, examine be the discovery that gas phase sample introduction needle needle core and syringe part twitch extremely glue it is puckery caused by, analysis may
Reason is larger (account for about whole ball 40%) for dragee coatings dosage, and including sucrose, along with sample is that water proposes acetic acid second
Ester extraction, then probably introduces a small amount of water soluble ingredient, such as sucrose in the sample, so that using only acetic acid second after sample introduction
Ester washes that needle is not thorough enough, occurs certain gluing puckery phenomenon after causing sample introduction needle number of injections more.
After considering above-mentioned reason, then determine to re-start test article treating method investigation after dragee to be removed to sugar-coat, and
Sample concentration is reduced as far as possible under instrumental sensitivity enabled condition.
6. taking dragee appropriate, sugar-coat is removed, fine powder is ground into, precision weighs 4g, adds water 25mL, is ultrasonically treated 30min, adds
Ethyl acetate extracts 2 times, each 20ml, and combining extraction liquid is recycled to dry, and residue adds ethyl acetate to redissolve, and is settled to 5ml appearance
In measuring bottle, shake up, cross 0.45 μm of miillpore filter to get.
7. taking dragee appropriate, sugar-coat is removed, fine powder is ground into, precision weighs 1g, adds water 25mL, is ultrasonically treated 30min, adds
Ethyl acetate extracts 2 times, first time 15ml, second of 10ml, and combining extraction liquid shakes up into 25ml volumetric flask, crosses 0.22 μm
Miillpore filter to get.
8. taking film-coating ball appropriate, it is ground into fine powder, precision weighs 1g, adds water 25mL, is ultrasonically treated 30min, adds acetic acid second
Ester extracts 2 times, first time 15ml, second of 10ml, and combining extraction liquid shakes up into 25ml volumetric flask, crosses 0.22 μm of micropore filter
Film to get.
As a result still there is sample introduction needle and glues puckery phenomenon after multiple sample introduction in the sample that 6. test article treating method obtains,
Therefore give up.
Test article treating method 7., 8. because sample concentration is lower, therefore sample introduction needle is more smooth after sample feeding, does not occur
Puckery phenomenon is glued, primarily determines using this method, carries out methodological study.The typical chromatogram of test article treating method 7., 8. is shown in
Figure 10, figure, 11.
The determination of 5 test article treating methods
Take female clever ball appropriate, dragee removes sugar-coat, is ground into fine powder, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, first time 15ml, second of 10ml, and combining extraction liquid shakes up into 25ml volumetric flask,
Cross 0.22 μm of miillpore filter to get.
6 chemical component qualitative analyses and the selection referring to peak
To further clarify ingredient in female clever ball, when GC-MS qualitative method is investigated using retention time of reference substance
The chemical component that may contain in female spirit ball.Pass through NIST8.0 in retention time between contrast sample and reference substance and GC-MS
Library comparison is composed, under the chromatographic condition, the chemical component that can be positioned in the sample has a Paeonol, α-cyperolone, in chromatogram
The content highest of Paeonol, and be the index components of content determination item in product, therefore select Paeonol for referring to peak.It is detailed in figure
12, Figure 13.
The preparation of Paeonol reference substance solution: taking Paeonol reference substance appropriate, accurately weighed, and ethyl acetate is added to be made often
1mL containing the solution to about 12.55 μ g of Paeonol to get.
The methodology validation of 7 female clever ball gas-phase fingerprint pattern tests
7.1 repetitive test
It takes with female clever ball (the film-coating ball lot number: 20130102 of a batch；Dragee lot number: 20130101), " 5 " Xiang Xiacao is pressed
Make, prepares 6 parts of test samples in parallel and analyzed using chromatographic condition under " 3 " item.
With the retention time of Paeonol and peak area for 1, the relative retention time of other each shared fingerprint peaks is calculated
(R.Rt) and the ratio of relative peak area (R.A).It the results are shown in Table 7-1~table 7-2.As a result repeatability is good.
Table 7-1 film-coating ball repeatability relative retention time, relative peak area tables of data
Table 7-2 dragee repeatability relative retention time, relative peak area tables of data
7.2 precision test
It takes with female clever ball (the film-coating ball lot number: 20130102 of a batch；Dragee lot number: 20130101), " 5 " Xiang Xiacao is pressed
Make, prepares 1 part of test sample continuous sample introduction 6 times, using chromatographic condition under " 3 " item, analyzed.
With the retention time of Paeonol and peak area for 1, the relative retention time of other each shared fingerprint peaks is calculated
(R.Rt) and the ratio of relative peak area (R.A).It the results are shown in Table 7-3~table 7-4.As a result except No. 4 peaks are because being not up to baseline separation
Outside relative peak area precision > 5%, remaining each peak relative retention time and the equal precision of relative peak area are good.
Table 7-3 film-coating ball precision relative retention time, relative peak area tables of data
Table 7-4 dragee precision relative retention time, relative peak area tables of data
7.3 stability test
It takes with female clever ball (the film-coating ball lot number: 20130102 of a batch；Dragee lot number: 20130101), " 5 " Xiang Xiacao is pressed
Make, prepare 1 part of test sample in 0,2,4,6,8,10,12,14,17,20,24H difference sample introduction, using chromatographic condition under " 3 " item, into
Row analysis.
With the retention time of Paeonol and peak area for 1, the relative retention time of other each shared fingerprint peaks is calculated
(R.Rt) and the ratio of relative peak area (R.A).It the results are shown in Table 7-5~table 7-6.As a result each peak relative retention time and opposite peak
Outside the equal < 5% of area RSD, show that sample stability is good in 24H.
Table 7-5 film-coating ball stability relative retention time, relative peak area tables of data
Table 7-6 dragee stability relative retention time, relative peak area tables of data
The analysis of 8 gas-phase fingerprint pattern methods
By female clever ball test article treating method determined above and chromatographic condition, 10 batches of female clever balls are randomly selected by " 5 " item
Lower operation is analyzed using chromatographic condition under " 3 " item.The foundation of 8.1 control gas-phase fingerprint patterns
Choose female clever ball film-coating 7 batches, dragee 3 batches, ball, by being operated under " 5 " item, using under chromatographic condition under " 3 " item into
Row measurement, according to relevant parameter given by 10 batches of female clever ball test sample gas phase maps, female spirit ball measures resulting main chromatography
Peak occurs in 45 minutes.Comparing film-coating ball, the chromatogram of 10 batches of samples of dragee and blank solvent chromatography graph discovery has
10 chromatographic peaks are each batch shared, see Figure 14.
Separately the relative retention time at 10 shared peaks in 10 batches of samples is analyzed, is to be calculated referring to peak with 5 Paeonols
The relative retention time at other 9 shared peaks, then the shared peak of 11 batches of samples is averaged relative retention time (peak number) successively are as follows:
0.365(1)、0.372(2)、0.769(3)、0.772(4)、1.000(5)、1.263(6)、1.283(7)、1.359(8)、1.395
(9), 1.565 (10), relative deviation is within ± 3%.It is shown in Table 8-1.
Table 8-110 batches of sample relative retention time tables of data
The fitting of 8.2 control gas-phase fingerprint patterns
It is shared peak with 10 peaks in the finger-print of above-mentioned 10 batches of samples, is commented using chromatographic fingerprints of Chinese materia medica similarity
Valence system (2.0 editions), fitting determine that female clever ball compares gas-phase fingerprint pattern, see Figure 15
8.3 the standard of finger-print limits
According to investigation above as a result, determining that the gas-phase fingerprint pattern similarity of female clever ball is as qualified 0.90 or more.
The beneficial effects of the present invention are:
The present invention has finally determined programmed temperature method to female spirit by attempting a variety of chromatographic conditions and test article treating method
Ball carries out the chromatographic condition of gas-phase fingerprint pattern analysis, and investigation has determined the control gas-phase fingerprint pattern at 10 shared peaks, method
It is easy, quick, reproducible, meet related request through methodology validation.
Using the gas-phase fingerprint pattern analysis method for the female clever ball of one kind that the present invention establishes, gas phase has been carried out to 10 batches of samples
As a result fingerprint map analyzing is compared with reference fingerprint and calculates similarity 0.90 or more, shows foundation of the present invention
This method be suitable for product quality control requirement.
The gas-phase fingerprint pattern analysis method strong applicability of the female clever ball of one kind that the present invention establishes, can more comprehensively reflect production
The quality condition of product.
Detailed description of the invention
Fig. 1, HP-5 capillary chromatographic column chromatogram
Fig. 2, DB-1701 capillary chromatographic column chromatogram
Fig. 3, DB-17 capillary chromatographic column chromatogram
Fig. 4, DB-1701 capillary chromatographic column chromatogram
Fig. 5, test article treating method 1. chromatogram
Fig. 6, test article treating method 2. chromatogram
Fig. 7, test article treating method 3. chromatogram
Fig. 8, test article treating method 4. chromatogram
Fig. 9, test article treating method 5. chromatogram
Figure 10, test article treating method 7. dragee chromatogram
Figure 11, test article treating method 8. film-coating ball chromatogram
Figure 12, Paeonol molecular ion peak figure
Figure 13, cyperolone molecular ion peak figure
Figure 14,10 batches of samples share peak and compare figure
Figure 15, female clever ball are fitted reference fingerprint
Specific embodiment:
The present invention is further illustrated by the following examples, but not as limitation of the present invention.
Embodiment 1, the gas-phase fingerprint pattern measurement of female spirit ball
Take female clever ball dragee, lot number 20120901
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 2, female spirit ball gas-phase fingerprint pattern measurement
Take female clever ball film-coating ball, lot number 20120809
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 3, female spirit ball gas-phase fingerprint pattern measurement
Take female clever ball dragee, lot number 20120903
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 4, female spirit ball gas-phase fingerprint pattern measurement
Chromatographic condition change are as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 6, the gas-phase fingerprint pattern measurement of female spirit ball
Take female clever ball dragee, lot number 20120901
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 0.8g, adds water 20mL, is ultrasonically treated
20min adds ethyl acetate to extract 2 times, the 1st 10ml, the 2nd 5ml, and combining extraction liquid adds acetic acid second into 15ml volumetric flask
Ester is settled to scale, shakes up, cross 0.2 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 150 DEG C；Detector: FID；Temperature: 230 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 0.5 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 7, the gas-phase fingerprint pattern measurement of female spirit ball
Take female clever ball dragee, lot number 20120901
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.2g, adds water 30mL, is ultrasonically treated
40min adds ethyl acetate to extract 2 times, the 1st 20ml, the 2nd 15ml, and combining extraction liquid adds acetic acid second into 35ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 200 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 5: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 8, the gas-phase fingerprint pattern measurement of female spirit ball
Take female clever ball dragee, lot number 20120903
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 9, the gas-phase fingerprint pattern measurement of female spirit ball
Take female clever ball dragee, lot number 20120901
Step 1, the preparation of test solution
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
Temperature programming table
Test solution is analyzed with this condition, obtains the finger-print of female clever ball to be measured.
Embodiment 10, the preparation of the female clever ball control gas-phase fingerprint pattern of standard
1) preparation of test solution, the method is as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get.
2) preparation of reference substance solution, the method is as follows:
Paeonol reference substance is taken, adds ethyl acetate that the solution that every 1mL contains 12.5 μ g is made, it is molten as object of reference reference substance
Liquid.
3) determination of finger-print, the method is as follows:
Test solution and control solution are injected into gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
1 temperature programming table of table
Analysis comparison is carried out to the gas-phase fingerprint pattern of 11 batches of female clever balls (7 batches, film-coating ball, dragee 4 batches), it is determined that
10 shared peaks, in chromatogram, chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, calculates other 9 altogether
There is the relative retention time at peak to be respectively as follows: 0.365 (1), 0.372 (2), 0.769 (3), 0.772 (4), 1.000 (5), 1.263
(6), 1.283 (7), 1.359 (8), 1.395 (9), 1.565 (10), the relative retention time at each peak is within ± 3%, in utilization
Medicine chromatographic fingerprinting similarity evaluation system (2.0 editions), fitting, which determines, obtains female clever ball standard control gas-phase fingerprint pattern.
Embodiment 11, the preparation of the female clever ball control gas-phase fingerprint pattern of standard
1) preparation of test solution, the method is as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs, and 0.8g adds water 20mL, ultrasonic treatment
20min adds ethyl acetate to extract 2 times, the 1st 10ml, the 2nd 5ml, and combining extraction liquid adds acetic acid second into 15ml volumetric flask
Ester is settled to scale, shakes up, cross 0.2 μm of miillpore filter to get.
2) preparation of reference substance solution, the method is as follows:
Paeonol reference substance is taken, adds ethyl acetate that the solution that every 1mL contains 10 μ g is made, as object of reference reference substance solution.
3) determination of finger-print, the method is as follows:
Test solution and control solution are injected into gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 150 DEG C；Detector: FID；Temperature: 230 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 0.5 μ l.
1 temperature programming table of table
Analysis comparison is carried out to the gas-phase fingerprint pattern of 11 batches of female clever balls (7 batches, film-coating ball, dragee 4 batches), it is determined that
10 shared peaks, in chromatogram, chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, calculates other 9 altogether
There is the relative retention time at peak to be respectively as follows: 0.365 (1), 0.372 (2), 0.769 (3), 0.772 (4), 1.000 (5), 1.263
(6), 1.283 (7), 1.359 (8), 1.395 (9), 1.565 (10), the relative retention time at each peak is within ± 3%, in utilization
Medicine chromatographic fingerprinting similarity evaluation system (2.0 editions), fitting, which determines, obtains female clever ball standard control gas-phase fingerprint pattern.
Embodiment 12, the preparation of the female clever ball control gas-phase fingerprint pattern of standard
1) preparation of test solution, the method is as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.2g, adds water 30mL, is ultrasonically treated
40min adds ethyl acetate to extract 2 times, the 1st 20ml, the 2nd 15ml, and combining extraction liquid adds acetic acid second into 35ml volumetric flask
Ester is settled to scale, shakes up, cross 0.3 μm of miillpore filter to get.
2) preparation of reference substance solution, the method is as follows:
Paeonol reference substance is taken, adds ethyl acetate that the solution that every 1mL contains 15 μ g is made, as object of reference reference substance solution.
3) determination of finger-print, the method is as follows:
Test solution and control solution are injected into gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 200 DEG C；Detector: FID；Temperature: 280 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 5: 1；Sample volume: 1 μ l.
1 temperature programming table of table
Analysis comparison is carried out to the gas-phase fingerprint pattern of 11 batches of female clever balls (7 batches, film-coating ball, dragee 4 batches), it is determined that
10 shared peaks, in chromatogram, chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, calculates other 9 altogether
There is the relative retention time at peak to be respectively as follows: 0.365 (1), 0.372 (2), 0.769 (3), 0.772 (4), 1.000 (5), 1.263
(6), 1.283 (7), 1.359 (8), 1.395 (9), 1.565 (10), the relative retention time at each peak is within ± 3%, in utilization
Medicine chromatographic fingerprinting similarity evaluation system (2.0 editions), fitting, which determines, obtains female clever ball standard control gas-phase fingerprint pattern.
Embodiment 13
Step 1, the preparation of female clever ball standard gas-phase fingerprint pattern, the method is as follows:
1) preparation of test solution, the method is as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get；
2) preparation of reference substance solution, the method is as follows:
Paeonol reference substance is taken, adds ethyl acetate that the solution that every 1mL contains 12.5 μ g is made, it is molten as object of reference reference substance
Liquid.
3) determination of finger-print, the method is as follows:
Test solution and control solution are injected into gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
1 temperature programming table of table
Analysis comparison is carried out to the gas-phase fingerprint pattern of 11 batches of female clever balls (7 batches, film-coating ball, dragee 4 batches), it is determined that
10 shared peaks, in chromatogram, chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, calculates other 9 altogether
There is the relative retention time at peak to be respectively as follows: 0.365 (1), 0.372 (2), 0.769 (3), 0.772 (4), 1.000 (5), 1.263
(6), 1.283 (7), 1.359 (8), 1.395 (9), 1.565 (10), the relative retention time at each peak is within ± 3%, in utilization
Medicine chromatographic fingerprinting similarity evaluation system (2.0 editions), fitting, which determines, obtains female clever ball standard control gas-phase fingerprint pattern.
The preparation method of female clever ball gas-phase fingerprint pattern to be measured, steps are as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 1.0g, adds water 25mL, is ultrasonically treated
30min adds ethyl acetate to extract 2 times, the 1st 15ml, the 2nd 10ml, and combining extraction liquid adds acetic acid second into 25ml volumetric flask
Ester is settled to scale, shakes up, cross 0.22 μm of miillpore filter to get；Solution is injected into gas chromatograph, obtains chromatogram.
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, (30m × 0.25mm ×
0.25μm).Column temperature: temperature programming condition see the table below；Injector temperature: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas:
Nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ l.
1 temperature programming table of table
Step 3, female clever ball gas-phase fingerprint pattern step 1 and step 2 obtained compares, and similarity is that 0.90 the above are conjunctions
Lattice.
Claims (9)
1. a kind of method for building up of female clever ball gas chromatography standard reference fingerprint, which comprises the steps of:
1) prepared by test solution:
Female clever ball is taken, adds water, ultrasonic extraction, extracting solution adds ethyl acetate to extract, and extract liquor takes ethyl acetate layer, adds ethyl acetate
Be settled to scale, shake up, cross miillpore filter to get；
2) gas chromatogram is obtained:
By test solution air inlet chromatography, gas chromatogram is obtained；
3) reference fingerprint is generated
Shared peak is determined in the gas-phase fingerprint pattern of more batches of qualified female clever ball samples of selection, utilizes chromatographic fingerprints of Chinese materia medica phase
Like degree evaluation system, determines female clever ball reference fingerprint, the standard reference material of effective component can be added as needed to make
For object of reference peak,
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase is using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, column temperature: temperature programming condition
It see the table below；Injector temperature: 150~200 DEG C；Detector: FID；Temperature: 230~280 DEG C；Carrier gas: nitrogen；Split sampling；Point
Stream ratio: 1: 1~5: 1；Sample volume: 0.5~1 μ l；
Temperature programming table
2. the method for building up of gas chromatography standard reference fingerprint according to claim 1, which is characterized in that including such as
Lower step:
1) preparation of test solution, the method is as follows:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 0.8~1.2g, adds 20~30mL of water, ultrasound
20~40min is handled, ethyl acetate is added to extract 2 times, the 1st 10~20ml, the 2nd 5~15ml, combining extraction liquid to 15~
In 35ml volumetric flask, ethyl acetate is added to be settled to scale, shaken up, cross 0.2-0.3 μm of miillpore filter to get；
2) gas chromatogram is obtained:
Standard reference material solution, method are prepared using Paeonol as object of reference are as follows: take Paeonol reference substance appropriate, add ethyl acetate
The solution that every 1mL contains 10~15 μ g is made, as reference substance solution；
Test solution and control solution are injected into gas chromatograph, obtain chromatogram, wherein chromatographic condition are as follows: chromatographic column is solid
Fixed phase is using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler；Column temperature: see temperature programming condition table；Injection port temperature
Degree: 180 DEG C；Detector: FID；Temperature: 250 DEG C；Carrier gas: nitrogen；Split sampling；Split ratio: 1: 1；Sample volume: 1 μ L；
3) reference fingerprint is generated
Analysis comparison is carried out to the gas-phase fingerprint pattern of ten batches or more female clever balls, determines 10 shared peaks, in chromatogram, with pellet
Retention time identical chromatographic peak in skin phenol reference substance peak is reference, and the relative retention time for calculating other 9 shared peaks is respectively as follows:
0.365,0.372,0.769,0.772,1.000,1.263,1.283,1.359,1.395,1.565, relative deviation is ±
3%, using similarity evaluation, fitting, which determines, obtains female clever ball standard control gas phase fingerprint image
Spectrum.
3. the method for building up of gas chromatography standard reference fingerprint according to claim 1, which is characterized in that 1) for examination
Product solution the preparation method is as follows:
Take female clever ball appropriate, dragee removes sugar-coat, is ground into fine powder, and precision weighs 1.0g, adds water 25mL, it is ultrasonically treated 30min,
Ethyl acetate is added to extract 2 times, first time 15ml, second of 10ml, combining extraction liquid adds ethyl acetate fixed into 25ml volumetric flask
Hold to scale, shake up, cross 0.22 μm of miillpore filter to get.
4. the method for building up of gas chromatography standard reference fingerprint according to claim 2, which is characterized in that 2) root bark of tree peony
Phenol reference substance solution the preparation method comprises the following steps: take Paeonol reference substance appropriate, it is accurately weighed, add ethyl acetate be made every 1mL containing pair
The solution of about 12.55 μ g of Paeonol to get.
5. the method for building up of gas chromatography standard reference fingerprint according to claim 1, which is characterized in that its intermediate range
Sequence heating table are as follows:
6. a kind of Gas Chromatography Fingerprint detection method of female clever ball, which comprises the steps of:
1) preparation of sample to be tested chromatogram:
Female clever ball is taken, adds water, ultrasonic extraction, extracting solution adds ethyl acetate to extract, and extract liquor takes ethyl acetate layer, adds ethyl acetate
It is settled to scale, is shaken up, miillpore filter is crossed, takes filtrate, gas chromatograph is injected, obtains chromatogram；
2) sample to be tested chromatogram and standard control fingerprint similarity calculate:
" similarity evaluation A editions " that the chromatogram of sample to be tested is imported Chinese Pharmacopoeia committee is soft
Part compares the chromatogram and standard control fingerprint similarity of sample to be tested；
Wherein the chromatographic condition of gas-chromatography is as follows:
Chromatographic column: stationary phase is using 14% cyanogen propvl-phenvl, 86% dimethyl polysiloxane as filler, column temperature: temperature programming condition
It see the table below；Injector temperature: 150~200 DEG C；Detector: FID；Temperature: 230~280 DEG C；Carrier gas: nitrogen；Split sampling；Point
Stream ratio: 1: 1~5: 1；Sample volume: 0.5~1 μ l；
Temperature programming table
7. Gas Chromatography Fingerprint detection method according to claim 6, which comprises the steps of:
1) preparation of sample to be tested chromatogram:
It takes female clever ball appropriate, is ground into fine powder, dragee removes sugar-coat, and precision weighs 0.8~1.2g, adds 20~30mL of water, ultrasound
20~40min is handled, ethyl acetate is added to extract 2 times, the 1st 10~20ml, the 2nd 5~15ml, combining extraction liquid to 15~
In 35ml volumetric flask, ethyl acetate is added to be settled to scale, shaken up, cross miillpore filter, take filtrate, injected gas chromatograph, obtain
Chromatogram；
2) sample to be tested chromatogram and standard control fingerprint similarity calculate:
The chromatogram of sample to be tested is imported to " similarity evaluation A editions " software of Chinese Pharmacopoeia committee
Compare the chromatogram and standard control fingerprint similarity of sample to be tested.
8. Gas Chromatography Fingerprint detection method according to claim 6, which is characterized in that wherein step 1) sample to be tested color
The preparation of spectrogram, the method is as follows:
Take female clever ball appropriate, dragee removes sugar-coat, is ground into fine powder, and precision weighs 1.0g, adds water 25mL, it is ultrasonically treated 30min,
Ethyl acetate is added to extract 2 times, first time 15ml, second of 10ml, combining extraction liquid adds ethyl acetate fixed into 25ml volumetric flask
Hold to scale, shake up, cross 0.22 μm of miillpore filter, take filtrate, injects gas chromatograph, obtain chromatogram.
9. Gas Chromatography Fingerprint detection method according to claim 6, which is characterized in that temperature programming table are as follows:
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