CN107402265A - The detection method of Kang Yuan particle finger-prints - Google Patents
The detection method of Kang Yuan particle finger-prints Download PDFInfo
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Abstract
The invention discloses the detection method of Kang Yuan particle finger-prints, Kang Yuan particles finger-print, which is established, to be comprised the following steps：The preparation of step 1, Kang Yuan particle need testing solutions；The preparation of step 2, reference substance solution：Step 3, accurate need testing solution of drawing injects liquid chromatograph respectively, records chromatogram；Step 3, the Kang Yuan particles finger-print obtained in step 2 is exported into instrument, and imports similarity evaluation, select existing chromatographic peak in the chromatogram of different batches Kang Yuan particles to be used as shared peak；With the reference fingerprint of mean value calculation method generation Kang Yuan particles；Calculate relative retention time, the relative peak area at each shared peak.Kang Yuan particles finger-print provided by the present invention, the quality of Kang Yuan particles comprehensively, can be objectively characterized, be advantageous to the quality of overall monitor medicine.The detection method of finger-print provided by the invention have method it is easy, stably, precision is high, high repeatability and other advantages.
Description
Technical field
The present invention relates to a kind of detection method of Chinese patent drug, and in particular to the detection method of Kang Yuan particle finger-prints.
Background technology
Finger-print refers to some complex materials, such as Chinese medicine, certain organism or the DNA of certain tissue or cell, egg
After white matter is appropriately processed, using certain analysis means, what is obtained can indicate the chromatogram or spectrogram of its chemical feature.
Traditional Chinese medicine fingerprint is a kind of synthesis, and quantifiable identification of means, it is built upon the base of chemical composition of Chinese materia medica system research
On plinth, it is mainly used in evaluating authenticity, Optimality and the stability of Chinese medicine and Chinese medicine preparation quality.Chinese medicine and its preparation are equal
For multi-component complex system, therefore evaluate its quality and should use adaptable therewith, the detection side for enriching authentication information can be provided
Method, establishing traditional Chinese medicine fingerprint more can will comprehensively reflect the species and quantity of contained chemical composition in Chinese medicine and its preparation,
And then whole description and evaluation are carried out to drug quality.
Kang Yuan particles are by the Radix Astragali, Radix Angelicae Sinensis, the root of herbaceous peony, the fruit of Chinese wolfberry, rhizoma cyperi (system), radix bupleuri, Poria cocos, teasel root, the bighead atractylodes rhizome (stir-fry), old
Compound preparation (Chinese medicines quasi-word Z32020996) made of skin, radix glycyrrhizae has soothing liver-qi stagnation, regulating qi-flowing for relieving pain, nourishing blood for regulating menstruation.For passing through
Early stage catatonic syndrome and primary dysmenorrhea, premenstrual swollen breasts, through carrying out underbelly pain.Said preparation medicinal material species is more, activity into
Divide complexity, the qualitative and quantitative analysis of individual components is difficult to the comprehensive information of reaction medicine comprehensively, for more effectively control Kang Yuan
The quality of grain, the present invention establishes the finger-print of preparation for Kang Yuan particle effective components, to comprehensive objective evaluation Kang Yuan
The quality of grain has great importance.
The content of the invention
Goal of the invention：Present invention aims to solve the deficiencies of the prior art, and provides a kind of a kind of finger-print of Kang Yuan particles
Detection method, the detection method can be with quality that is objective, comprehensively and accurately evaluating Kang Yuan particles, the matter to controlling Kang Yuan particles
Amount and guarantee clinical efficacy are significant.
Technical scheme：To achieve these goals, the technical scheme taken of the present invention is：
A kind of detection method of Kang Yuan particles finger-print, comprises the following steps：
The preparation of step 1, Kang Yuan particle need testing solutions：
The Kang Yuan particles of different batches are taken, it is accurately weighed, put in conical flask, precision absorption petroleum ether solution, ultrasonic degreasing,
Taking filter residue to add methanol solution after filtration, weigh, ultrasonic extraction, weighed weight after cooling, difference weight supplied with methanol solution,
Subsequent filtrate is taken, obtains need testing solution；
The preparation of step 2, reference substance solution：
Accurately weighed Paeoniflorin reference substance, forulic acid reference substance, liquiritin reference substance and aurantiamarin reference substance, are placed in not
In same volumetric flask, add methanol solution and dissolve and be settled to graduation mark, respectively obtain Paeoniflorin reference substance solution, forulic acid pair
According to product solution, liquiritin reference substance solution and aurantiamarin reference substance solution；
Step 3, respectively accurate absorption need testing solution and reference substance solution, inject high performance liquid chromatograph, record chromatogram
Figure；
Step 4, the finger-print of the Kang Yuan particle need testing solutions obtained in step 3 is exported, and imports Chinese medicine chromatogram
Fingerprint similarity evaluation system；Existing chromatographic peak is as shared in the chromatogram of selection different batches Kang Yuan particles
Peak；With the reference fingerprint of mean value calculation method generation Kang Yuan particles, the relative retention time at each shared peak and relative is calculated
Peak area；And the chemical composition at peak in reference fingerprint is marked according to the retention time of reference substance solution chromatogram.
Preferably, the detection method of above-described Kang Yuan particles finger-print, step 1 Kang Yuan particle test samples
Solution manufacturing method is：Kang Yuan particles each 17.0g of 10 batches is taken, it is accurately weighed, put in 100mL conical flasks, precision is drawn
20mL petroleum ether solutions, ultrasonic 30min degreasings, take filter residue to add methanol solution 20mL after filtration, weigh, ultrasonic extraction 30min,
Weighed weight after cooling, difference weight are supplied with methanol solution, are taken subsequent filtrate, are obtained need testing solution.
Preferably, the detection method of above-described Kang Yuan particles finger-print, the system of step 2 reference substance solution
It is standby：A certain amount of Paeoniflorin reference substance, forulic acid reference substance, liquiritin reference substance, aurantiamarin reference substance are taken, it is accurately weighed, point
It is not placed in different volumetric flasks, adds methanol solution and dissolve and be settled to graduation mark, it is 0.2064mg/mL that concentration is made respectively
Paeoniflorin reference substance solution, 0.1242mg/mL forulic acids reference substance solution, 0.2695mg/mL liquiritins reference substance solution and
0.1026mg/mL aurantiamarin reference substance solution.
Preferably, the detection method of above-described Kang Yuan particles finger-print, in step 3, liquid chromatogram bar
Part is：Chromatographic column：Chromatographic column is YMC-Pack-ODS-A C18 chromatographic columns；Mobile phase A is 0.05% phosphate aqueous solution, flowing
Phase B is acetonitrile, gradient elution, program 0-10min, 5%-10%B；10-20min, 10%-15%B；20-40min,
15%-21%B；40-60min, 21%-25%B；60-80 min, 25%-30%B；Detection wavelength：230nm；Column temperature：30℃；
Flow velocity：0.6mL/min；Sample size：10μL.
Preferably, the detection method of above-described Kang Yuan particles finger-print, selection 9 is altogether in finger-print
There is characteristic peak of the peak as Kang Yuan particle finger-prints, using the peak area of this 9 characteristic peaks as the variable index of quality evaluation,
Paeoniflorin is No. 6 peaks in 9 shared peaks, and retention time is 42.167 minutes, forulic acid is No. 7 peaks, retention time 49.589
Minute, liquiritin are No. 8 peaks, and retention time is 50.586 minutes, aurantiamarin is No. 9 peaks, and retention time is 62.633 minutes.
Beneficial effect：
1st, the present invention is according to active component contained in Kang Yuan particles, including the chemical combination such as saponins, organic acid and flavonoids
The structural property feature of thing, optimal flowing phase composition is filtered out by many experiments, gradient elution program, flow velocity, detects ripple
The long, analysis condition such as chromatographic column, column temperature, show through many experiments checking, Kang Yuan particles finger-print detection provided by the invention
Method, can it is comprehensive, objective, accurate detection and evaluation Kang Yuan particles quality, for ensure clinical efficacy it is significant.
2nd, the Kang Yuan particle finger-prints established with method provided by the present invention, can effectively characterize Kang Yuan sheet of particles
Quality, objective can embody each tandem and correlation for forming fingerprint characteristic peak, focus on overall facial feature, both may be used
Avoid judging because determining individual chemical composition the one-sidedness of Kang Yuan granular mass, can be reduced to requisite quality again and artificially handle
Possibility.
3rd, the detection method of Kang Yuan particles finger-print provided by the invention, with method is easy, stability is good, precision
High, high repeatability and other advantages.
Brief description of the drawings
Fig. 1 is the reference fingerprint of Kang Yuan particles of the present invention.
Fig. 2 is 10 batch test sample finger-prints of Kang Yuan particles of the present invention.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, unreceipted actual conditions in embodiment
Person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, being can be with
Pass through the conventional products of acquisition purchased in market.
The instrument that embodiment is used and reagent are as follows：
1. experiment material and instrument and equipment
1.1 sample：This experiment have collected 10 batches of Kang Yuan particulate samples altogether, respectively from Nanjing Hydron Chinese medicine group (H1,
H2, H3, H4, H5) and Heilungkiang Ji Ren pharmaceutcal corporation, Ltds (J6, J7, J8, J9, J10).
1.2 reagents and reagent
Reference substance：Paeoniflorin (lot number：170109), forulic acid (lot number：161204), liquiritin (lot number:161216) it is, orange
Skin glycosides (lot number:161230) all it is purchased from Nanjing Sen Beijia bio tech ltd.
Reagent：Petroleum ether (Nanjing Chemistry Reagent Co., Ltd.), analysis methanol (Nanjing Chemistry Reagent Co., Ltd.), phosphoric acid
(Nanjing Chemistry Reagent Co., Ltd.)；Chromatogram acetonitrile (U.S. world company)；Make distilled water by oneself.
1.3 experimental instrument and equipment
Shimadzu LC-20AB highly effective liquid phase chromatographic systems (Japanese Shimadzu Corporation, including on-line degassing machine, auto injection
Device, PDAD, column oven).BP121S electronic analytical balances (SARTORIUS)；KQ5200DB type ultrasonic waves are clear
Wash device (Kunshan Ultrasonic Instruments Co., Ltd.).
A kind of detection method of Kang Yuan particles finger-print of embodiment 1, comprises the following steps：
The preparation of step 1, Kang Yuan particle need testing solutions：
The Kang Yuan particles of the above 10 batches each 17.0g is taken, it is accurately weighed, put in 100mL conical flasks, precision draws 20mL oil
Ethereal solution, ultrasonic 30min degreasings, methanol solution 20mL is added after filtration, is weighed, ultrasonic extraction 30min is weighed heavy after cooling
Amount, difference weight are supplied with methanol solution, take subsequent filtrate to obtain.
Step 2, the preparation with reference to peak solution：
A certain amount of Paeoniflorin reference substance, forulic acid reference substance, liquiritin reference substance, aurantiamarin reference substance are taken, precision claims
It is fixed, it is respectively placed in different volumetric flasks, adds methanol solution and dissolve and be settled to graduation mark, concentration is made respectively is
0.2064mg/mL Paeoniflorins reference substance solution, 0.1242mg/mL forulic acids reference substance solution, the control of 0.2695mg/mL liquiritins
The aurantiamarin reference substance solution of product solution and 0.1026mg/mL；
Step 3, respectively accurate absorption need testing solution and reference substance solution, inject high performance liquid chromatograph, record chromatogram
Figure；Chromatographic condition is：Chromatographic column：Chromatographic column is using YMC-Pack-ODS-AC18 (250mm × 4.6 mm, 5 μm)；Mobile phase A
(0.05% phosphate aqueous solution)-B (acetonitrile), gradient elution, program 0-10min, 5%-10%B；10-20min, 10%-
15%B；20-40min, 15%-21%B；40-60min, 21%-25% B；60-80min, 25%-30%B；Detection wavelength：
230nm；Column temperature：30℃；Flow velocity：0.6mL/min；Sample size：10μL.
Step 4, the finger-print of the Kang Yuan particle need testing solutions obtained in step 3 is exported, and imports Chinese medicine chromatogram
Fingerprint similarity evaluation system；Existing chromatographic peak is as shared in the chromatogram of selection different batches Kang Yuan particles
Peak；With the reference fingerprint of mean value calculation method generation Kang Yuan particles, the relative retention time at each shared peak and relative is calculated
Peak area；Characteristic peak of 9 shared peaks as Kang Yuan particle finger-prints is have selected on the finger-print of Kang Yuan particles, with this
Variable index of the peak area of 9 characteristic peaks as quality evaluation.Meanwhile using reference substance counter point to main in finger-print
Characteristic peak is demarcated, and as a result 4 characteristic peaks are accredited, and respectively Paeoniflorin, forulic acid, liquiritin, aurantiamarin, chromatogram are shown in
Fig. 1.Because being shown in Kang Yuan particulate chromatography figures, paeoniflorin content is higher, therefore the reference peak using Paeoniflorin as Kang Yuan particles
(S).Using the retention time at S peaks and peak area as 1, by the retention time of other each characteristic peaks and peak area and S in same collection of illustrative plates
The retention time and peak area at peak make ratio, calculate the relative retention time and relative peak area value of other each characteristic peaks, as a result
See Tables 1 and 2.
The Kang Yuan particles of table 1 share peak relative retention time
| Numbering | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| H1 | 0.33 | 0.64 | 0.74 | 0.88 | 0.92 | 1.00 | 1.19 | 1.20 | 1.49 |
| H2 | 0.33 | 0.64 | 0.74 | 0.88 | 0.92 | 1.00 | 1.18 | 1.20 | 1.49 |
| H3 | 0.33 | 0.63 | 0.72 | 0.88 | 0.90 | 1.00 | 1.18 | 1.20 | 1.48 |
| H4 | 0.31 | 0.62 | 0.72 | 0.87 | 0.90 | 1.00 | 1.18 | 1.20 | 1.49 |
| H5 | 0.34 | 0.64 | 0.75 | 0.90 | 0.94 | 1.00 | 1.20 | 1.24 | 1.51 |
| J6 | 0.33 | 0.65 | 0.75 | 0.90 | 0.93 | 1.00 | 1.20 | 1.24 | 1.53 |
| J7 | 0.33 | 0.65 | 0.75 | 0.90 | 0.93 | 1.00 | 1.20 | 1.24 | 1.53 |
| J8 | 0.33 | 0.65 | 0.75 | 0.89 | 0.94 | 1.00 | 1.19 | 1.21 | 1.48 |
| J9 | 0.34 | 0.66 | 0.76 | 0.90 | 0.93 | 1.00 | 1.19 | 1.20 | 1.48 |
| J10 | 0.34 | 0.67 | 0.75 | 0.90 | 0.94 | 1.00 | 1.19 | 1.20 | 1.49 |
| Average value | 0.33 | 0.65 | 0.75 | 0.89 | 0.93 | 1.00 | 1.19 | 1.21 | 1.50 |
| RSD (%) | 2.17 | 2.28 | 1.86 | 1.21 | 1.77 | 0.00 | 0.60 | 1.39 | 1.32 |
The Kang Yuan particles of table 2 share peak relative peak area
| Numbering | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| H1 | 0.30 | 0.70 | 0.10 | 0.26 | 0.28 | 1.00 | 0.05 | 0.05 | 0.29 |
| H2 | 0.30 | 0.72 | 0.09 | 0.27 | 0.29 | 1.00 | 0.05 | 0.05 | 0.29 |
| H3 | 0.31 | 0.71 | 0.10 | 0.27 | 0.29 | 1.00 | 0.05 | 0.05 | 0.26 |
| H4 | 0.40 | 0.88 | 0.08 | 0.29 | 0.37 | 1.00 | 0.07 | 0.05 | 0.31 |
| H5 | 0.38 | 0.73 | 0.10 | 0.28 | 0.34 | 1.00 | 0.07 | 0.05 | 0.37 |
| J6 | 0.33 | 0.69 | 0.07 | 0.27 | 0.32 | 1.00 | 0.06 | 0.05 | 0.34 |
| J7 | 0.33 | 0.74 | 0.09 | 0.22 | 0.31 | 1.00 | 0.07 | 0.05 | 0.30 |
| J8 | 0.38 | 0.77 | 0.10 | 0.23 | 0.31 | 1.00 | 0.07 | 0.06 | 0.32 |
| J9 | 0.33 | 0.80 | 0.11 | 0.28 | 0.28 | 1.00 | 0.06 | 0.06 | 0.30 |
| J10 | 0.39 | 0.87 | 0.07 | 0.27 | 0.33 | 1.00 | 0.06 | 0.05 | 0.29 |
| Average value | 0.35 | 0.76 | 0.09 | 0.26 | 0.31 | 1.00 | 0.06 | 0.05 | 0.30 |
| RSD (%) | 10.61 | 8.20 | 11.97 | 9.44 | 8.31 | 1.00 | 12.13 | 5.61 | 9.64 |
The HPLC data of the Kang Yuan particles of 10 batches are exported as into AIA forms, importing Chinese Pharmacopoeia Commission, " Chinese medicine refers to
Line collection of illustrative plates similarity evaluation system 2004 editions ", standard finger-print is generated, and select 9 chromatographic peaks to be characterized peak, carry out fingerprint
Collection of illustrative plates similarity evaluation.The finger-print common pattern of 10 samples is shown in Fig. 2, and similarity evaluation the results are shown in Table 3.According to fingerprint image
Spectrum similarity evaluation result shows that the Kang Yuan particles of different batches have certain difference, and result of study is shown, finger-print phase
The Kang Yuan particles that can be used for distinguishing different batches like degree analysis, can be used for the quality control of Kang Yuan particles.
The different batches Kang Yuan particle fingerprint similarities of table 3
Methodological study
1st, precision test
Reference substance solution under accurate extraction embodiment 1, by the chromatographic condition continuous sample introduction 6 times of embodiment 1 time, it is mainly common
The RSD for having peak relative retention time is respectively less than 0.35%；Relative peak area RSD is respectively less than 1.99%, shows the precision of instrument
Well.Specific experiment the results are shown in Table 4 and table 5.
The relative retention time of table 4
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 13.31 | 27.69 | 31.72 | 36.42 | 38.19 | 41.44 | 49.12 | 49.73 | 61.53 |
| 2 | 13.24 | 27.65 | 31.68 | 36.42 | 38.19 | 41.46 | 49.11 | 49.75 | 61.52 |
| 3 | 13.24 | 27.66 | 31.67 | 36.41 | 38.17 | 41.44 | 49.08 | 49.72 | 61.50 |
| 4 | 13.22 | 27.62 | 31.64 | 36.39 | 38.15 | 41.42 | 49.08 | 49.71 | 61.47 |
| 5 | 13.17 | 27.61 | 31.63 | 36.42 | 38.18 | 41.47 | 49.11 | 49.79 | 61.61 |
| 6 | 13.20 | 27.60 | 31.63 | 36.38 | 38.14 | 41.41 | 49.03 | 49.70 | 61.46 |
| Average value | 13.23 | 27.64 | 31.66 | 36.41 | 38.17 | 41.44 | 49.09 | 49.73 | 61.51 |
| RSD (%) | 0.35 | 0.13 | 0.11 | 0.05 | 0.06 | 0.05 | 0.06 | 0.07 | 0.09 |
The relative peak area of table 5
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 1447239 | 3384972 | 477458 | 1259902 | 1433427 | 4430880 | 279689 | 209589 | 1371959 |
| 2 | 1453970 | 3355351 | 475318 | 1229348 | 1456993 | 4540866 | 277006 | 209627 | 1361977 |
| 3 | 1439072 | 3370799 | 479211 | 1235616 | 1484320 | 4524718 | 278058 | 204130 | 1375078 |
| 4 | 1441271 | 3373013 | 478505 | 1232541 | 1421485 | 4401632 | 281058 | 208797 | 1380261 |
| 5 | 1491642 | 3396725 | 468591 | 1273559 | 1459636 | 4539816 | 280270 | 205158 | 1386724 |
| 6 | 1465891 | 3373644 | 469617 | 1287415 | 1497991 | 4537334 | 278053 | 204691 | 1386290 |
2nd, stability test
Same need testing solution is taken, is measured respectively in 0h, 2h, 4h, 8h, 12h, 24h sample introduction, it mainly shares peak phase
0.89% is respectively less than to the RSD of retention time；Relative peak area RSD is respectively less than 2.76%, the results showed that：Need testing solution exists
It is stable in 24h.Specific experiment the results are shown in Table 6 and table 7.
The relative retention time of table 6
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 13.75 | 27.45 | 31.95 | 37.00 | 38.11 | 41.98 | 49.73 | 50.24 | 62.07 |
| 2 | 13.72 | 27.58 | 31.32 | 36.90 | 38.20 | 41.58 | 49.32 | 49.89 | 61.72 |
| 3 | 13.67 | 27.81 | 31.24 | 36.94 | 38.23 | 41.53 | 49.25 | 49.82 | 61.60 |
| 4 | 13.58 | 27.85 | 31.63 | 36.61 | 38.85 | 41.71 | 49.39 | 50.00 | 61.78 |
| 5 | 13.91 | 27.19 | 31.69 | 37.22 | 38.81 | 41.63 | 49.30 | 49.90 | 61.68 |
| 6 | 13.82 | 27.50 | 31.78 | 37.26 | 38.22 | 41.47 | 49.08 | 49.76 | 61.54 |
| Average value | 13.74 | 27.56 | 31.60 | 36.99 | 38.40 | 41.65 | 49.35 | 49.94 | 61.73 |
| RSD (%) | 0.82 | 0.89 | 0.86 | 0.64 | 0.87 | 0.43 | 0.44 | 0.34 | 0.30 |
The relative peak area of table 7
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 1493011 | 3316083 | 424913 | 1237054 | 1490853 | 4321788 | 263565 | 207108 | 1331371 |
| 2 | 1457487 | 3450787 | 437558 | 1218680 | 1456793 | 4245384 | 292594 | 211275 | 1319836 |
| 3 | 1479082 | 3319605 | 415866 | 1229713 | 1414081 | 4341488 | 282205 | 208934 | 1314851 |
| 4 | 1499768 | 3288822 | 417118 | 1259633 | 1436381 | 4244591 | 277146 | 211241 | 1319762 |
| 5 | 1449453 | 3170446 | 414142 | 1224445 | 1456987 | 4249257 | 284743 | 216808 | 1329033 |
| 6 | 1492398 | 3263020 | 421475 | 1201783 | 1480624 | 4241995 | 291338 | 201927 | 1323894 |
| Average value | 1478533 | 3301461 | 421845 | 1228551 | 1455953 | 4274084 | 286514 | 209549 | 1321475 |
| RSD (%) | 1.40 | 2.76 | 2.05 | 1.57 | 1.93 | 1.05 | 1.89 | 2.37 | 0.40 |
3rd, replica test
Precision weighs same crowd of sample 17.0g, totally 6 parts, according to the lower section legal system available test sample solution of embodiment 1, to implement
Chromatographic condition sample introduction under example 1.It mainly shares the RSD of peak relative retention time and is respectively less than 0.80%；Relative peak area RSD
Respectively less than 2.97%, show that the repeatability of method is good.Specific experiment the results are shown in Table 8 and table 9.
The relative retention time of table 8
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 13.85 | 27.17 | 31.27 | 37.03 | 38.88 | 41.98 | 49.73 | 50.24 | 62.07 |
| 2 | 13.74 | 27.04 | 31.17 | 36.94 | 38.78 | 41.86 | 49.62 | 50.16 | 62.03 |
| 3 | 13.86 | 27.14 | 31.24 | 37.05 | 38.87 | 41.62 | 49.38 | 49.94 | 61.74 |
| 4 | 13.55 | 27.30 | 31.33 | 37.04 | 38.81 | 41.53 | 49.26 | 49.87 | 61.74 |
| 5 | 13.73 | 27.39 | 31.41 | 37.07 | 38.83 | 41.44 | 49.12 | 49.73 | 61.53 |
| 6 | 13.75 | 27.27 | 31.32 | 37.12 | 38.77 | 41.43 | 49.03 | 49.71 | 61.45 |
| Average value | 13.75 | 27.22 | 31.29 | 37.04 | 38.82 | 41.64 | 49.36 | 49.94 | 61.76 |
| RSD (%) | 0.80 | 0.46 | 0.27 | 0.16 | 0.12 | 0.55 | 0.56 | 0.44 | 0.41 |
The relative peak area of table 9
| Sequence number | Peak 1 | Peak 2 | Peak 3 | Peak 4 | Peak 5 | Peak 6 | Peak 7 | Peak 8 | Peak 9 |
| 1 | 1434069 | 3351623 | 429217 | 1221647 | 1452549 | 4321788 | 263565 | 206108 | 1291371 |
| 2 | 1470525 | 3382806 | 433880 | 1284361 | 1478797 | 4379005 | 267321 | 201496 | 1297137 |
| 3 | 1490332 | 3313236 | 438533 | 1250777 | 1472463 | 4412850 | 264299 | 202017 | 1291829 |
| 4 | 1435673 | 3382993 | 421941 | 1237950 | 1497964 | 4492724 | 275338 | 215441 | 1335969 |
| 5 | 1411458 | 3218307 | 427703 | 1183041 | 1429111 | 4430880 | 279689 | 209589 | 1311959 |
| 6 | 1434696 | 3201401 | 443975 | 1219582 | 1457294 | 4500138 | 260172 | 201723 | 1300306 |
| Average value | 1446126 | 3308394 | 432542 | 1232893 | 1464696 | 4422898 | 269364 | 204367 | 1308312 |
| RSD (%) | 1.99 | 2.44 | 1.84 | 2.76 | 1.63 | 1.54 | 2.97 | 2.22 | 1.84 |
Test result indicates that, Kang Yuan particles fingerprint atlas detection method provided by the invention, stability is good, precision above
Height, reproducible, the quality of the comprehensive objective evaluation Kang Yuan particles of energy, to ensure that clinical efficacy has great importance.
Above example is only the exemplary embodiment of the present invention, is not used in the limitation present invention, protection scope of the present invention
It is defined by the claims.Those skilled in the art can make respectively in the essence and protection domain of the present invention to the present invention
Kind modification or equivalent substitution, this modification or equivalent substitution also should be regarded as being within the scope of the present invention.
Claims (5)
1. a kind of detection method of Kang Yuan particles finger-print, it is characterised in that comprise the following steps：
The preparation of step 1, Kang Yuan particle need testing solutions：
The Kang Yuan particles of different batches are taken, it is accurately weighed, put in conical flask, precision draws petroleum ether solution, ultrasonic degreasing, filtration
After take filter residue to add methanol solution, weigh, ultrasonic extraction, weighed weight after cooling, difference weight supplied with methanol solution, taken continuous
Filtrate, obtain need testing solution；
The preparation of step 2, reference substance solution：
Accurately weighed Paeoniflorin reference substance, forulic acid reference substance, liquiritin reference substance and aurantiamarin reference substance, are placed in different
In volumetric flask, add methanol solution and dissolve and be settled to graduation mark, respectively obtain Paeoniflorin reference substance solution, forulic acid reference substance
Solution, liquiritin reference substance solution and aurantiamarin reference substance solution；
Step 3, respectively accurate absorption need testing solution and reference substance solution, inject high performance liquid chromatograph, record chromatogram；
Step 4, the finger-print of the Kang Yuan particle need testing solutions obtained in step 3 is exported, and imports Chinese medicine chromatographic fingerprint
Collection of illustrative plates similarity evaluation system；Existing chromatographic peak in the chromatogram of different batches Kang Yuan particles is selected to be used as shared peak；With
Mean value calculation method generates the reference fingerprint of Kang Yuan particles, calculates the relative retention time at each shared peak and relative peak face
Product；And the chemical composition at peak in reference fingerprint is marked according to the retention time of reference substance solution chromatogram.
2. the detection method of Kang Yuan particles finger-print according to claim 1, it is characterised in that step 1 Kang Yuan particles
Need testing solution preparation method is：Kang Yuan particles each 17.0g of 10 batches is taken, it is accurately weighed, put in 100mL conical flasks, it is accurate
20mL petroleum ether solutions are drawn, ultrasonic 30min degreasings, takes filter residue to add methanol solution 20mL after filtration, weighs, ultrasonic extraction
30min, weighed weight after cooling, difference weight are supplied with methanol solution, take subsequent filtrate, obtain need testing solution.
3. the detection method of Kang Yuan particles finger-print according to claim 1, it is characterised in that step 2 reference substance is molten
The preparation of liquid：A certain amount of Paeoniflorin reference substance, forulic acid reference substance, liquiritin reference substance, aurantiamarin reference substance are taken, precision claims
It is fixed, it is respectively placed in different volumetric flasks, adds methanol solution and dissolve and be settled to graduation mark, concentration is made respectively is
0.2064mg/mL Paeoniflorins reference substance solution, 0.1242mg/mL forulic acids reference substance solution, the control of 0.2695mg/mL liquiritins
The aurantiamarin reference substance solution of product solution and 0.1026mg/mL.
4. the detection method of Kang Yuan particles finger-print according to claim 1, it is characterised in that in step 3, liquid phase color
Spectral condition is：Chromatographic column：Chromatographic column is YMC-Pack-ODS-A C18 chromatographic columns；Mobile phase A is 0.05% phosphate aqueous solution,
Mobile phase B is acetonitrile, gradient elution, program 0-10min, 5%-10%B；10-20min, 10%-15%B；20-40min,
15%-21%B；40-60min, 21%-25%B；60-80min, 25%-30%B；Detection wavelength：230nm；Column temperature：30℃；
Flow velocity：0.6mL/min；Sample size：10μL.
5. the detection method of Kang Yuan particles finger-print according to claim 1, it is characterised in that selected in finger-print
Characteristic peak of 9 shared peaks as Kang Yuan particle finger-prints, the variable of quality evaluation is used as using the peak area of this 9 characteristic peaks
Index, Paeoniflorin is No. 6 peaks in 9 shared peaks, and retention time is 42.167 minutes, forulic acid is No. 7 peaks, and retention time is
49.589 minutes, liquiritin be No. 8 peaks, retention time is 50.586 minutes, aurantiamarin is No. 9 peaks, retention time 62.633
Minute.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109490438A (en) * | 2018-11-13 | 2019-03-19 | 南京海昌中药集团有限公司 | The detection method of Wumiaoshuixiangao-multi-effect ointment finger-print |
| CN111879884A (en) * | 2020-07-06 | 2020-11-03 | 武汉同济现代医药科技股份有限公司 | Quality control method of Chinese medicinal preparation |
| CN114942291A (en) * | 2022-06-20 | 2022-08-26 | 江苏省中医院 | Method for detecting quality of 'Zhenyang Yangyin' granule |
| CN115798573A (en) * | 2023-02-02 | 2023-03-14 | 武汉宏韧生物医药股份有限公司 | Method and device for analyzing medicinal components |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101816773A (en) * | 2010-01-27 | 2010-09-01 | 湖南中医药大学 | Method for measuring content of superfine powder preparation in five retentions powder |
| CN103191278A (en) * | 2013-04-01 | 2013-07-10 | 江苏海昇药业有限公司 | Chinese medicinal composition for treating premenstrual tension syndrome and preparation thereof |
-
2017
- 2017-07-05 CN CN201710542907.4A patent/CN107402265B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101816773A (en) * | 2010-01-27 | 2010-09-01 | 湖南中医药大学 | Method for measuring content of superfine powder preparation in five retentions powder |
| CN103191278A (en) * | 2013-04-01 | 2013-07-10 | 江苏海昇药业有限公司 | Chinese medicinal composition for treating premenstrual tension syndrome and preparation thereof |
Non-Patent Citations (4)
| Title |
|---|
| 孙立华 等: "高效液相色谱法测定康媛颗粒中阿魏酸的含量", 《中国药事》 * |
| 杜晔 等: "RP-HPLC法同时测定柴归消癥胶囊中6种成分", 《中成药》 * |
| 潘萍 等: "康媛颗粒质量标准研究", 《中国中医药信息杂志》 * |
| 王晓玲 等: "HPLC法同时测定香连化滞丸中10种成分", 《中草药》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109490438A (en) * | 2018-11-13 | 2019-03-19 | 南京海昌中药集团有限公司 | The detection method of Wumiaoshuixiangao-multi-effect ointment finger-print |
| CN111879884A (en) * | 2020-07-06 | 2020-11-03 | 武汉同济现代医药科技股份有限公司 | Quality control method of Chinese medicinal preparation |
| CN114942291A (en) * | 2022-06-20 | 2022-08-26 | 江苏省中医院 | Method for detecting quality of 'Zhenyang Yangyin' granule |
| CN114942291B (en) * | 2022-06-20 | 2024-03-12 | 江苏省中医院 | Quality detection method of yang-suppressing yin-nourishing particles |
| CN115798573A (en) * | 2023-02-02 | 2023-03-14 | 武汉宏韧生物医药股份有限公司 | Method and device for analyzing medicinal components |
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