CN105486764A - Gas-phase fingerprint analysis method for Kunling Wan and application thereof - Google Patents
Gas-phase fingerprint analysis method for Kunling Wan and application thereof Download PDFInfo
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Abstract
The invention provides a gas-phase fingerprint analysis method for Kunling Wan. The method comprises the following steps of establishment of a standard fingerprint spectrum of the Kunling Wan, determination of the fingerprint spectrum of Kunling Wan to-be-determined, and comparison and determination of similarity. According to the invention, by trying a plurality of program heating conditions, the method finally determines the chromatographic condition for gas-phase fingerprint spectrum analysis of the Kunling Wan, investigates contrast gas-phase fingerprint spectrums of ten common peaks, and has the characteristics of simple process, fast speed, and good reappearance, the established standard gas-phase fingerprint spectrum can satisfy relative requirements through methodology validation, applicability is strong, quality status of the product can be comprehensively reflected, and the method provides an important basis for quality controlling and standard making.
Description
Technical field
The present invention relates to a kind of gas-phase fingerprint pattern analytical approach of female clever ball, belong to Pharmaceutical Analysis technical field.
Background technology
Female clever ball is a kind of compound Chinese medicinal preparation, and its function cures mainly as menstruation regulating blood nourishing, raw new by the stasis of blood.For irregular menstruation, more or less, menalgia, cold uterus, fails to be impregnated for a long time, habitual abortion, leukorrhea with reddish discharge, and uterine bleeding is more than, the sick deficiency of vital energy for a long time, deficiency of the kidney pain in the back.
The prescription of female clever ball is as follows:
[prescription]
(1) rhizoma cyperi (system) the 37g Radix Glycyrrhizae 7g radix cynanchi atrati 14g motherwort 14g Radix Astragali 14g cockscomb 14g 14g tuber of dwarf lilyturf fructus schisandrae 14g glutinous rehmannia 14g safflower 14g akebi 10g bighead atractylodes rhizome (stir-fry) 14g red halloysite 14g Poria cocos 14g bark of official magnolia 10g saline cistanche (system) the 14g root of herbaceous peony (wine stir-fry) 14g
(2) rhizoma cyperi (system) 37g schizonepeta 10g moutan bark 14g donkey-hide gelatin 14g Radix Angelicae Sinensis 14g Jehol Ligusticum Rhizome 10g red ginseng 14g deer horn glue 14g Bulbus Fritillariae Cirrhosae 14g myrrh (stir-fry) 14g fructus amomi 14g corydalis tuber 14g fennel seeds (salt system) 14g colla carapacis et plastri testudinis 14g Ligusticum wallichii 14g
[method for making] above prescription (one) 17 taste, boiling secondary, 3 hours first times, second time 2 hours, collecting decoction, filter, filtrate places 24 hours, gets supernatant and adds rice vinegar 280g, be condensed into thick paste.Above prescription (two) ten five tastes, are ground into middle powder, add in above-mentioned thick paste, and mixing, dry, pulverize into fine powder, sieve, and mixing, with the general ball of yellow rice wine, makes 1800 balls, sugar coating or film-coating, polishing, dry, to obtain final product.
Because chemical composition of Chinese materia medica is complicated, mechanism of action is indefinite, existing herbal pharmaceutical technology is caused to be difficult to guarantee tcm product quality stability, the kind that comprehensively can reflect Chinese medicine intrinsic chemical composition that traditional Chinese medicine fingerprint is set up and quantity, and then the quality of reflection Chinese medicine.In present stage, the effective elements of the medicine overwhelming majority is not clear and definite, adopts the mode of traditional Chinese medicine fingerprint, effectively will characterize traditional Chinese medicine quality.Finger-print is approved by international community, is conducive to Chinese medicine and product introduction international market.
Female clever ball act.std 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 related in standard, at present, female clever ball is not carried out to the report of finger-print research, and finger-print fully can characterize globality and the complicacy of Chinese medicine preparation, therefore the effective constituent of inventor to female clever ball is studied.Find wherein containing multiple water-soluble and Alcohol soluble composition, the present inventor is studied these compositions targetedly, establish a kind of standard finger-print of female clever ball, utilize this collection of illustrative plates to carry out quality supervision, to guarantee product quality to the female clever ball in production.
Summary of the invention:
The invention provides a kind of female clever ball gas chromatography standard reference fingerprint, its method for building up is as follows:
1) need testing solution preparation:
Get female clever ball, add water, ultrasonic extraction, extract adds extraction into ethyl acetate, and extract ethyl acetate layer adds ethyl acetate and is settled to scale, shakes up, and crosses 0.22 μm of miillpore filter, both;
2) gas chromatogram is obtained:
Need testing solution gas chromatograph, obtains gas chromatogram;
3) reference fingerprint is generated
Select to determine 10 total peaks in the gas-phase fingerprint pattern of many batches of qualified female clever ball samples, utilize similarity evaluation (2.0 editions), female clever ball reference fingerprint is determined in matching.
The standard reference material that can add effective constituent is as required using as object of reference peak.
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition is in table 1; Injector temperature: 150 ~ 200 DEG C; Detecting device: FID; Temperature: 230 ~ 280 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1 ~ 5: 1; Sample size: 0.5 ~ 1 μ l.
Table 1 temperature programme table
Temperature programme table is preferred:
Wherein, the method for building up of preferred standard control finger-print is as follows:
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 0.8 ~ 1.2g, and add water 20 ~ 30mL, ultrasonic process 20 ~ 40min, add extraction into ethyl acetate 2 times, 1st 10 ~ 20ml, the 2nd 5 ~ 15ml, in combining extraction liquid to 15 ~ 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product;
Preferably, get female clever ball appropriate, dragee removing sugar-coat, be ground into fine powder, precision takes 1.0g, add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
2) gas chromatogram is obtained:
Need testing solution gas chromatograph, obtains gas chromatogram;
Wherein using Paeonol as object of reference preparation standard reference substance solution, method is as follows:
Get Paeonol reference substance appropriate, add ethyl acetate and make the solution of every 1mL containing 10 ~ 15 μ g, product solution in contrast, excellent to get Paeonol reference substance appropriate, adds ethyl acetate and make every 1mL containing the solution to Paeonol about 12.55 μ g, to obtain final product.
3) generate reference fingerprint, method is as follows:
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram,
Com-parison and analysis is carried out to the gas-phase fingerprint pattern of many batches batches of female clever balls, determine 10 total peaks, in chromatogram, the chromatogram identical with Paeonol reference substance peak retention time is No. 5 peaks with reference to peak, the relative retention time calculating other 9 total peaks is respectively: 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), its relative deviation is all within ± 3%, utilize similarity evaluation (2.0 editions), matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
The present invention also provides a kind of Gas Chromatography Fingerprint detection method of female clever ball, it is characterized in that, comprises the steps
1) preparation of testing sample chromatogram:
Get female clever ball, add water, ultrasonic extraction, extract adds extraction into ethyl acetate, and extract ethyl acetate layer adds ethyl acetate and is settled to scale, shakes up, and crosses 0.22 μm of miillpore filter, gets filtrate, inject gas chromatograph, obtain chromatogram;
2) testing sample chromatogram and standard control fingerprint similarity calculate:
The chromatogram of testing sample is imported " similarity evaluation A version " software of Chinese Pharmacopoeia committee, compare chromatogram and the standard control fingerprint similarity of testing sample;
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition is in table 1; Injector temperature: 150 ~ 200 DEG C; Detecting device: FID; Temperature: 230 ~ 280 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1 ~ 5: 1; Sample size: 0.5 ~ 1 μ l.
Table 1 temperature programme table
Temperature programme table is preferred:
Wherein, preferred detection method of the present invention is as follows:
1) preparation of testing sample chromatogram:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 0.8 ~ 1.2g, and add water 20 ~ 30mL, ultrasonic process 20 ~ 40min, add extraction into ethyl acetate 2 times, the 1st 10 ~ 20ml, the 2nd 5 ~ 15ml, in combining extraction liquid to 15 ~ 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, get filtrate, inject gas chromatograph, obtain chromatogram;
Preferably, get female clever ball appropriate, dragee removing sugar-coat, be ground into fine powder, precision takes 1.0g, add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
2) testing sample chromatogram and standard control fingerprint similarity calculate:
The chromatogram of testing sample is imported " similarity evaluation A version " software of Chinese Pharmacopoeia committee, compare chromatogram and the standard control fingerprint similarity of testing sample; Similarity is more than 0.90 for qualified, and similarity is more than 0.95 for conforming to, and similarity is more than 0.98 for identical.
The present invention is by the foundation of above finger-print, further establish the analytical approach of female clever ball, adopt the method can the female clever ball of accurate discrimination, according to the comparison with standard finger-print, draw discriminating conclusion, more and standard finger-print close, product quality is better, and similarity can use computer approach to confirm, is qualified if similarity is more than 0.90, similarity is more than 0.95 for conforming to, and similarity is more than 0.98 for identical.
The preparation method of the said method particularly standard control gas-phase fingerprint pattern of female clever ball is through that screening obtains, and is below screening process:
1 instrument and reagent
Instrument: U.S. Agilent7890A, AgilentOpenLABControlPanel workstation.
Chromatographic column: 1. non-polar column, Stationary liquid with 5% phenyl 95% dimethyl polysiloxane for filler, AgilentHP-5 (30m × 0.32mm × 0.25 μm);
2. Semi-polarity post, Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, AgilentDB-1701 (30m × 0.25mm × 0.25 μm).
3. Semi-polarity post, Stationary liquid 50% diphenyl 50% dimethyl polysiloxane, AgilentDB-17 (30m × 0.25mm × 0.15 μm)
Reagent: ethyl acetate (chromatographically pure, Tianjin Concord Technology Co., Ltd.); Water is ultrapure water.
Reference substance: Paeonol (assay is used for Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110708-200506); α-cyperolone (assay is used for National Institute for Food and Drugs Control, lot number: 110748-201111); Honokiol (assay use, by 98.4% purity meter for National Institute for Food and Drugs Control, lot number: 110730-201011)
Sample: female clever ball (Tianjin Tasly (Liaoning) Pharmaceutical Co., Ltd.), totally 11 batches, concrete lot number, in table 3, is all up to the standards through Product Quality Verification Centers.
Table 3 sample message table
The selection of 2 GC conditions
The selection of 2.1 chromatographic columns
Because female clever ball comprises dragee and film-coating ball two kinds of specifications, and dragee sugar-coat agent weightening finish large percentage (about 40%), therefore (lot number: 20130209) sample carries out carries out methodological study for dragee and film-coating ball again after condition is determined substantially first to adopt plain ball during method investigation.
With reference to pertinent literature, investigate opposed polarity capillary column to test, as a result 3 kinds of opposed polarity capillary columns be separated the chromatographic peak number that obtains and peak shape basically identical, (see Fig. 1 ~ 3), but wherein chromatographic column chromatographic peak peak shape is 2. better, therefore chromatographic column is used 2. to carry out the investigation of temperature programme condition.
The selection of 2.2 temperature programme conditions
Use chromatographic column 2. to carry out investigations adjustment to temperature programme condition, ensureing out peak completely in situation, shorten analysis duration as far as possible.Result, after have adjusted more than 30 secondary program Elevated Temperature Conditions, determines that isolate about 11 chromatographic peaks altogether, analysis time, overall length was 55min, saw Fig. 4 under this temperature programme condition (see table 2-1).
Table 2-1 temperature programme table
The determination of 3 GC conditions
To sum up analyze, finally determine that female clever ball gas-phase fingerprint pattern chromatographic condition is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, AgilentDB-1701 (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition is in table 1; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ L.
Table 1 temperature programme table
The selection of 4 test sample disposal routes
First 4.1 adopt dragee to carry out the selection of Extraction solvent:
1. get dragee appropriate, be ground into fine powder, precision takes 7g (converting with plain ball), adds ethyl acetate 25mL, ultrasonic process 30min, centrifuging and taking supernatant, is concentrated into dry, and residue adds ethyl acetate and redissolves and be settled to 5ml, shake up, cross 0.45 μm of miillpore filter, to obtain final product.
2. get dragee appropriate, be ground into fine powder, precision takes 7g (converting with plain ball), adds cyclohexane 25mL, ultrasonic process 30min, centrifuging and taking supernatant, is concentrated into dry, and residue adds cyclohexane and redissolves and be settled to 5ml, shake up, cross 0.45 μm of miillpore filter, to obtain final product.
3. get dragee appropriate, be ground into fine powder, precision takes 7g, and add water 25mL, ultrasonic process 30min, add cyclohexane and extract 2 times, each 20ml, combining extraction liquid is recycled to dry, and residue adds cyclohexane and redissolves, and be settled in 5ml volumetric flask, shake up, cross 0.45 μm of miillpore filter, to obtain final product.
4. get dragee appropriate, be ground into fine powder, precision takes 7g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, each 20ml, combining extraction liquid is recycled to dry, and residue adds ethyl acetate and redissolves, and be settled in 5ml volumetric flask, shake up, cross 0.45 μm of miillpore filter, to obtain final product.
The results are shown in Figure 5 ~ Fig. 8, as seen from the figure, adopt test sample disposal route to be 4. that the chromatogram information that water extraction extraction into ethyl acetate obtains is maximum, therefore determine to adopt test sample disposal route 4., continue to investigate.
4.2 according to above-mentioned investigation result, adopts dragee to carry out the selection of extracting mode, and namely 5. to get dragee appropriate for test sample disposal route, be ground into fine powder, precision takes 7g, and add water 25mL, add hot reflux 30min, add extraction into ethyl acetate 2 times, each 20ml, combining extraction liquid is recycled to dry, residue adds ethyl acetate and redissolves, and is settled in 5ml volumetric flask, shakes up, cross 0.45 μm of miillpore filter, to obtain final product.
The results are shown in Figure 9, as seen from the figure, except 3 chromatographic peaks of retention time before 10min obviously become greatly, all the other chromatographic peaks all obviously diminish, therefore consider the globality of gas-phase fingerprint pattern information and the simplicity of test sample operation, select test sample disposal route 4..
4. 4.3 adopt above-mentioned test sample disposal route, carry out in dragee methodological study process, there is the phenomenon of gas phase sample introduction needle bending, examine and find it is that gas phase sample introduction needle nook closing member and syringe part twitch very sticky puckery causing, analyzing possible cause is dragee coatings large usage quantity (accounting for 40% of whole ball), and comprise sucrose, adding sample is water extraction extraction into ethyl acetate, the water soluble ingredient that then very possible introducing is a small amount of in the sample to which, as sucrose etc., only ethyl acetate is adopted to wash pin after making sample introduction thorough not, cause sample introduction needle number of injections more after there is certain sticky puckery phenomenon.
After considering above-mentioned reason, then re-start test sample disposal route after determining that dragee is removed sugar-coat and investigate, and reduce sample concentration as far as possible under instrumental sensitivity enabled condition.
6. get dragee appropriate, removing sugar-coat, is ground into fine powder, precision takes 4g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, each 20ml, combining extraction liquid is recycled to dry, and residue adds ethyl acetate and redissolves, and be settled in 5ml volumetric flask, shake up, cross 0.45 μm of miillpore filter, to obtain final product.
7. get dragee appropriate, removing sugar-coat, be ground into fine powder, precision takes 1g, and add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
8. get film-coating ball appropriate, be ground into fine powder, precision takes 1g, and add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, and combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
The sample that 6. result test sample disposal route obtains, after repeatedly sample introduction, has still occurred that sample introduction needle glues puckery phenomenon, therefore has given up.
Test sample disposal route 7., 8. because sample concentration is lower, therefore after sample feeding, sample introduction needle is comparatively smooth, does not occur sticky puckery phenomenon, tentatively determines to adopt the method, carries out methodological study.Test sample disposal route typical color spectrogram 7., is 8. shown in Figure 10, figure, 11.
The determination of 5 test sample disposal routes
Get female clever ball appropriate, dragee removing sugar-coat, be ground into fine powder, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
6 chemical composition qualitative analyses and the selection with reference to peak
For ingredient in clearly female clever ball further, reference substance retention time is adopted to investigate the chemical composition that may contain in female clever ball to when GC-MS qualitative method.Storehouse contrast is composed by NIST8.0 in the retention time between comparative sample and reference substance and GC-MS, under this chromatographic condition, the chemical composition can located in the sample to which has Paeonol, α-cyperolone, in chromatogram, the content of Paeonol is the highest, and be the index components of assay item in product, therefore Paeonol is selected to be with reference to peak.Refer to Figure 12, Figure 13.
The preparation of Paeonol reference substance solution: get Paeonol reference substance appropriate, accurately weighed, add ethyl acetate and make every 1mL containing the solution to Paeonol about 12.55 μ g, to obtain final product.
The Method validation of 7 female clever ball gas-phase fingerprint pattern tests
7.1 replica test
Get with a collection of female clever ball (film-coating ball lot number: 20130102; Dragee lot number: 20130101), operates by under " 5 " item, parallel preparation 6 parts of test samples, and chromatographic condition under employing " 3 " item, analyzes.
With the retention time of Paeonol and peak area for 1, calculate the relative retention time (R.Rt) of other each total fingerprint peaks and the ratio of relative peak area (R.A).The results are shown in Table 7-1 ~ table 7-2.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
Get with a collection of female clever ball (film-coating ball lot number: 20130102; Dragee lot number: 20130101), operates by under " 5 " item, prepares 1 part of test sample continuous sample introduction 6 times, and chromatographic condition under employing " 3 " item, analyzes.
With the retention time of Paeonol and peak area for 1, calculate the relative retention time (R.Rt) of other each total fingerprint peaks and the ratio of relative peak area (R.A).The results are shown in Table 7-3 ~ table 7-4.Result except No. 4 peaks are not because reaching except baseline separation relative peak area precision > 5%, all the other each peak relative retention times and the equal precision of relative peak area 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
Get with a collection of female clever ball (film-coating ball lot number: 20130102; Dragee lot number: 20130101), by operating under " 5 " item, prepare 1 part of test sample in 0,2,4,6,8,10,12,14,17,20,24H sample introduction respectively, chromatographic condition under employing " 3 " item, analyzes.
With the retention time of Paeonol and peak area for 1, calculate the relative retention time (R.Rt) of other each total fingerprint peaks and the ratio of relative peak area (R.A).The results are shown in Table 7-5 ~ table 7-6.Outside result each peak relative retention time and the equal < 5% of relative peak area RSD, show that in 24H, sample stability is good.
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
8 gas-phase fingerprint pattern methods analysts
By the female clever ball test sample disposal route determined above and chromatographic condition, the female clever ball of random selecting 10 batches operates by under " 5 " item, and chromatographic condition under employing " 3 " item, analyzes.The foundation of 8.1 contrast gas-phase fingerprint patterns
Choose female clever 7 batches, ball film-coating ball, dragee 3 batches, operate by under " 5 " item, measure under chromatographic condition under employing " 3 " item, according to the correlation parameter given by 10 batches of female clever ball test sample gas phase collection of illustrative plates, the main chromatographic peak that female clever ball measures gained all occurred in 45 minutes.Relatively the chromatogram of film-coating ball, dragee 10 batch sample and blank solvent chromatogram find that there is 10 chromatographic peaks be each batch total, see Figure 14.
Separately the relative retention time at 10 total peaks in 10 batch samples is analyzed, with 5 Paeonols for reference peak, calculate the relative retention time at other 9 total peaks, then total peak average relative retention time (peak number) of 11 batch samples is followed successively by: 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), and its relative deviation is all within ± 3%.In Table 8-1.
Table 8-110 batch sample relative retention time tables of data
The matching of 8.2 contrast gas-phase fingerprint patterns
For total peak, utilize similarity evaluation (2.0 editions) with 10 peaks in the finger-print of above-mentioned 10 batch samples, female clever ball contrast gas-phase fingerprint pattern is determined in matching, sees Figure 15
The standard of 8.3 finger-prints limits
According to investigating result above, determine that the gas-phase fingerprint pattern similarity of female clever ball is more than 0.90 qualified.
Beneficial effect of the present invention is:
The present invention is by attempting plurality of color spectral condition and test sample disposal route, finally determine programmed temperature method carries out gas-phase fingerprint pattern analysis chromatographic condition to female clever ball, investigate the contrast gas-phase fingerprint pattern determining 10 total peaks, method is easy, quick, reproducible, meets related request through Method validation.
The gas-phase fingerprint pattern analytical approach of a kind of female clever ball that application the present invention sets up, gas-phase fingerprint pattern analysis has been carried out to 10 batch samples, result and reference fingerprint are compared and are calculated similarity all more than 0.90, show that the method that the present invention sets up is applicable to the quality control requirement of product.
The gas-phase fingerprint pattern analytical approach applicability of a kind of female clever ball that the present invention sets up is strong, can reflect the quality condition of product more comprehensively.
Accompanying drawing explanation
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 sample disposal route 1. chromatogram
Fig. 6, test sample disposal route 2. chromatogram
Fig. 7, test sample disposal route 3. chromatogram
Fig. 8, test sample disposal route 4. chromatogram
Fig. 9, test sample disposal route 5. chromatogram
Figure 10, test sample disposal route 7. dragee chromatogram
Figure 11, test sample disposal route 8. film-coating ball chromatogram
Figure 12, Paeonol molecular ion peak figure
Figure 13, cyperolone molecular ion peak figure
Figure 14,10 batch samples have peak comparison diagram
Figure 15, female clever ball matching reference fingerprint
Embodiment:
Further illustrate the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1, the gas-phase fingerprint pattern of female clever ball measures
Get female clever ball dragee, lot number is 20120901
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 2, female clever ball gas-phase fingerprint pattern measures
Get female clever ball film-coating ball, lot number 20120809
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 3, female clever ball gas-phase fingerprint pattern measures
Get female clever ball dragee, lot number 20120903
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 4, female clever ball gas-phase fingerprint pattern measures
Chromatographic condition changes to:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 6, the gas-phase fingerprint pattern of female clever ball measures
Get female clever ball dragee, lot number is 20120901
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 0.8g, and add water 20mL, ultrasonic process 20min, add extraction into ethyl acetate 2 times, 1st 10ml, the 2nd 5ml, combining extraction liquid is in 15ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.2 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 150 DEG C; Detecting device: FID; Temperature: 230 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 0.5 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 7, the gas-phase fingerprint pattern of female clever ball measures
Get female clever ball dragee, lot number is 20120901
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.2g, and add water 30mL, ultrasonic process 40min, add extraction into ethyl acetate 2 times, 1st 20ml, the 2nd 15ml, combining extraction liquid is in 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 200 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 5: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 8, the gas-phase fingerprint pattern of female clever ball measures
Get female clever ball dragee, lot number is 20120903
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 9, the gas-phase fingerprint pattern of female clever ball measures
Get female clever ball dragee, lot number is 20120901
Step 1, the preparation of need testing solution
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
Step 2, the foundation of gas-phase fingerprint pattern
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Temperature programme table
Analyze need testing solution with this understanding, obtain the finger-print of female clever ball to be measured.
Embodiment 10, the preparation of the female clever ball contrast gas-phase fingerprint pattern of standard
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product.
2) preparation of reference substance solution, method is as follows:
Get Paeonol reference substance, add ethyl acetate and make the solution of every 1mL containing 12.5 μ g, as object of reference reference substance solution.
3) determination of finger-print, method is as follows:
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Table 1 temperature programme table
To 11 batches of female clever ball (7 batches, film-coating balls, dragee 4 batches) gas-phase fingerprint pattern carry out com-parison and analysis, determine 10 total peaks, in chromatogram, the chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, the relative retention time calculating other 9 total peaks is respectively: 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%, utilize similarity evaluation (2.0 editions), matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
Embodiment 11, the preparation of the female clever ball contrast gas-phase fingerprint pattern of standard
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes, 0.8g, and add water 20mL, ultrasonic process 20min, adds extraction into ethyl acetate 2 times, the 1st 10ml, 2nd 5ml, combining extraction liquid, in 15ml volumetric flask, adds ethyl acetate and is settled to scale, shake up, cross 0.2 μm of miillpore filter, to obtain final product.
2) preparation of reference substance solution, method is as follows:
Get Paeonol reference substance, add ethyl acetate and make the solution of every 1mL containing 10 μ g, as object of reference reference substance solution.
3) determination of finger-print, method is as follows:
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 150 DEG C; Detecting device: FID; Temperature: 230 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 0.5 μ l.
Table 1 temperature programme table
To 11 batches of female clever ball (7 batches, film-coating balls, dragee 4 batches) gas-phase fingerprint pattern carry out com-parison and analysis, determine 10 total peaks, in chromatogram, the chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, the relative retention time calculating other 9 total peaks is respectively: 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%, utilize similarity evaluation (2.0 editions), matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
Embodiment 12, the preparation of the female clever ball contrast gas-phase fingerprint pattern of standard
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.2g, and add water 30mL, ultrasonic process 40min, add extraction into ethyl acetate 2 times, 1st 20ml, the 2nd 15ml, combining extraction liquid is in 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.3 μm of miillpore filter, to obtain final product.
2) preparation of reference substance solution, method is as follows:
Get Paeonol reference substance, add ethyl acetate and make the solution of every 1mL containing 15 μ g, as object of reference reference substance solution.
3) determination of finger-print, method is as follows:
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 200 DEG C; Detecting device: FID; Temperature: 280 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 5: 1; Sample size: 1 μ l.
Table 1 temperature programme table
To 11 batches of female clever ball (7 batches, film-coating balls, dragee 4 batches) gas-phase fingerprint pattern carry out com-parison and analysis, determine 10 total peaks, in chromatogram, the chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, the relative retention time calculating other 9 total peaks is respectively: 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%, utilize similarity evaluation (2.0 editions), matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
Embodiment 13
Step 1, the preparation of female clever ball standard gas-phase fingerprint pattern, method is as follows:
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product;
2) preparation of reference substance solution, method is as follows:
Get Paeonol reference substance, add ethyl acetate and make the solution of every 1mL containing 12.5 μ g, as object of reference reference substance solution.
3) determination of finger-print, method is as follows:
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram,
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Table 1 temperature programme table
To 11 batches of female clever ball (7 batches, film-coating balls, dragee 4 batches) gas-phase fingerprint pattern carry out com-parison and analysis, determine 10 total peaks, in chromatogram, the chromatographic peak identical with Paeonol reference substance peak retention time is No. 5 peaks, the relative retention time calculating other 9 total peaks is respectively: 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%, utilize similarity evaluation (2.0 editions), matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
The preparation method of female clever ball gas-phase fingerprint pattern to be measured, step is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, 1st 15ml, the 2nd 10ml, combining extraction liquid is in 25ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.22 μm of miillpore filter, to obtain final product; By solution inject gas chromatograph, obtain chromatogram.
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, (30m × 0.25mm × 0.25 μm).Column temperature: temperature programme condition sees the following form; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ l.
Table 1 temperature programme table
Step 3, female clever ball gas-phase fingerprint pattern step 1 and step 2 obtained compares, and similarity is more than 0.90 is qualified.
Claims (9)
1. a female clever ball gas chromatography standard reference fingerprint, its method for building up is as follows:
1) need testing solution preparation:
Get female clever ball, add water, ultrasonic extraction, extract adds extraction into ethyl acetate, and extract gets ethyl acetate layer, adds ethyl acetate and is settled to scale, shake up, and crosses miillpore filter, both;
2) gas chromatogram is obtained:
By need testing solution air inlet chromatography, obtain gas chromatogram;
3) reference fingerprint is generated
Select to determine total peak in the gas-phase fingerprint pattern of many batches of qualified female clever ball samples, utilize similarity evaluation, determine female clever ball reference fingerprint, the standard reference material that can add effective constituent is as required using as object of reference peak
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, column temperature: temperature programme condition sees the following form; Injector temperature: 150 ~ 200 DEG C; Detecting device: FID; Temperature: 230 ~ 280 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1 ~ 5: 1; Sample size: 0.5 ~ 1 μ l;
Temperature programme table
。
2. gas chromatography standard reference fingerprint according to claim 1, its method for building up is as follows:
1) preparation of need testing solution, method is as follows:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 0.8 ~ 1.2g, and add water 20 ~ 30mL, ultrasonic process 20 ~ 40min, add extraction into ethyl acetate 2 times, 1st 10 ~ 20ml, the 2nd 5 ~ 15ml, in combining extraction liquid to 15 ~ 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross 0.2-0.3 μm of miillpore filter, to obtain final product; 2) gas chromatogram is obtained:
Using Paeonol as object of reference preparation standard reference substance solution, method is: get Paeonol reference substance appropriate, adds ethyl acetate and makes the solution of every 1mL containing 10 ~ 15 μ g, product solution in contrast;
By need testing solution and reference substance solution inject gas chromatograph, obtain chromatogram, wherein chromatographic condition is: chromatographic column fixed phase with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler; Column temperature: see temperature programme condition table; Injector temperature: 180 DEG C; Detecting device: FID; Temperature: 250 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1; Sample size: 1 μ L;
3) reference fingerprint is generated
Com-parison and analysis is carried out to the gas-phase fingerprint pattern of more than ten batches female clever balls, determine 10 total peaks, in chromatogram, with the chromatographic peak identical with Paeonol reference substance peak retention time for reference, the relative retention time calculating other 9 total peaks is respectively: 0.365,0.372,0.769,0.772,1.000,1.263,1.283,1.359,1.395,1.565, its relative deviation is ± and 3%, utilize similarity evaluation, matching is determined to obtain female clever ball standard control gas-phase fingerprint pattern.
3. gas chromatography standard reference fingerprint according to claim 1, it is 1 years old) preparation method of need testing solution is as follows:
Get female clever ball appropriate, dragee removing sugar-coat, be ground into fine powder, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, adds extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid, in 25ml volumetric flask, shakes up, and crosses 0.22 μm of miillpore filter, to obtain final product.
4. gas chromatography standard reference fingerprint according to claim 2, they are 2 years old) preparation method of Paeonol reference substance solution is: get Paeonol reference substance appropriate, accurately weighed, add ethyl acetate and make every 1mL containing the solution to Paeonol about 12.55 μ g, to obtain final product.
5. gas chromatography standard reference fingerprint according to claim 1, wherein temperature programme table is:
。
6. a Gas Chromatography Fingerprint detection method for female clever ball, is characterized in that, comprise the steps:
1) preparation of testing sample chromatogram:
Get female clever ball, add water, ultrasonic extraction, extract adds extraction into ethyl acetate, and extract gets ethyl acetate layer, adds ethyl acetate and is settled to scale, shake up, and crosses miillpore filter, gets filtrate, inject gas chromatograph, obtain chromatogram;
2) testing sample chromatogram and standard control fingerprint similarity calculate:
The chromatogram of testing sample is imported " similarity evaluation A version " software of Chinese Pharmacopoeia committee, compare chromatogram and the standard control fingerprint similarity of testing sample;
Wherein the chromatographic condition of gas chromatography is as follows:
Chromatographic column: Stationary liquid with 14% cyanogen propvl-phenvl 86% dimethyl polysiloxane for filler, column temperature: temperature programme condition sees the following form; Injector temperature: 150 ~ 200 DEG C; Detecting device: FID; Temperature: 230 ~ 280 DEG C; Carrier gas: nitrogen; Split sampling; Split ratio: 1: 1 ~ 5: 1; Sample size: 0.5 ~ 1 μ l;
Temperature programme table
。
7. Gas Chromatography Fingerprint detection method according to claim 6, is characterized in that, comprise the steps:
1) preparation of testing sample chromatogram:
Get female clever ball appropriate, be ground into fine powder, dragee removing sugar-coat, precision takes 0.8 ~ 1.2g, and add water 20 ~ 30mL, ultrasonic process 20 ~ 40min, add extraction into ethyl acetate 2 times, the 1st 10 ~ 20ml, the 2nd 5 ~ 15ml, in combining extraction liquid to 15 ~ 35ml volumetric flask, add ethyl acetate and be settled to scale, shake up, cross miillpore filter, get filtrate, inject gas chromatograph, obtain chromatogram;
2) testing sample chromatogram and standard control fingerprint similarity calculate:
The chromatogram of testing sample is imported chromatogram and the standard control fingerprint similarity of " similarity evaluation A version " comparison testing sample of Chinese Pharmacopoeia committee.
8. Gas Chromatography Fingerprint detection method according to claim 6, is characterized in that, wherein step 1) preparation of testing sample chromatogram, method is as follows:
Get female clever ball appropriate, dragee removing sugar-coat, is ground into fine powder, precision takes 1.0g, and add water 25mL, ultrasonic process 30min, add extraction into ethyl acetate 2 times, first time 15ml, second time 10ml, combining extraction liquid is in 25ml volumetric flask, shake up, cross 0.22 μm of miillpore filter, get filtrate, inject gas chromatograph, obtains chromatogram.
9. Gas Chromatography Fingerprint detection method according to claim 6, is characterized in that, temperature programme table is:
。
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| CN114814059A (en) * | 2022-04-27 | 2022-07-29 | 天津中新药业集团股份有限公司隆顺榕制药厂 | Agilawood depression relieving tablet gas chromatography standard control fingerprint spectrum, construction method and application |
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