CN101797360B - Fingerprint detection method for analgesic plaster - Google Patents
Fingerprint detection method for analgesic plaster Download PDFInfo
- Publication number
- CN101797360B CN101797360B CN201010126328XA CN201010126328A CN101797360B CN 101797360 B CN101797360 B CN 101797360B CN 201010126328X A CN201010126328X A CN 201010126328XA CN 201010126328 A CN201010126328 A CN 201010126328A CN 101797360 B CN101797360 B CN 101797360B
- Authority
- CN
- China
- Prior art keywords
- retention time
- peaks
- relatively
- phase
- relative retention
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a fingerprint detection method for an analgesic plaster preparation. Chromatographic conditions of the method comprise that: a mobile phase is formed by taking octadecyl bonds and silica gel as filling agents, taking tetrahydrofuran-0.5 percent phosphoric acid as a phase A and taking acetonitrile as a phase B; a gradient elution mode is adopted; a detection wavelength is 3502nm; a flow rate is 0.5 to 1.5ml/min; a column temperature is between 20 and 50 DEG C; and a sample inlet volume is between 1 and 25 mu l. The fingerprint totally has 18 characteristic fingerprint peaks of which the peak area summation is over 90 percent of a total peak area; a No.10 chromatographic peak is a characteristic peak of luteolin of which the relative retention time and the relative peak area are both 1; and a standard fingerprint of an analgesic plaster is obtained by calculating the relative retention time and the relative peak area of other peaks. The fingerprint detection method for the analgesic plaster has the advantages of simple and convenient method, stability, high precision and high reproducibility; the quality situation of the analgesic plaster can be grasped from the overall appearance of the chromatograph; and the standard fingerprint can be used as an index for controlling the quality of the analgesic plaster.
Description
Technical field
The present invention relates to the fingerprint atlas detection method of Chinese medicine preparation, be specifically related to the detection method of the finger-print of analgesic plaster medicinal powder or cream or extract.
Background technology
Analgesic plaster is the tibetan traditional medicine that Tibet Linzhi's Qizheng Tibetan Medicine joint-stock company produces; This medicine adopts the Qinghai-Tibet distinctive natural drug of China; Form through special cryogenic vacuum freeze-dry process is refining, major function is promoting blood circulation and removing blood stasis, swelling and pain relieving, is used for acute and chronic and dampens, falls and beat bruise, osteoproliferation, rheumatism and rheumatoid pain; Enjoy great prestige both at home and abroad with its remarkable effect, rank first of the Chinese distinctive elaboration Tibetan medicine.
At present, its quality control index is except measuring content of luteolin, and other has lamiophlomis rotata, ginger-colored thin layer to differentiate; Though ability control of quality; But can't control quality condition comprehensively, for remedying above deficiency, make that its Quality Control Technology is more perfect, science and a kind of available quality control pattern is provided its standardized production from now on; 11 batches of products of quantity-produced have been carried out analyzing relatively, set up the assay method and the standard finger-print of the standard finger-print of this product extract.
Summary of the invention
The accurate fingerprint atlas detection method that the purpose of this invention is to provide a kind of analgesic plaster extract or preparation, this method have that method is easy, stable, precision is high, favorable reproducibility, the characteristics that are easy to grasp.
Chinese patent has been number for ZL200510080696.4 discloses analgesic plaster extract or preparation,
Raw material by following parts by weight is processed medicament:
Lamiophlomis rotata 5-15 part, whin 2-10 part, turmeric 15-60 part, Chinese prickly ash 7-20 part, cornu bubali 5-15 part and German tamarisk 15-40 part;
The preparation method is: Chinese prickly ash, Cornu Bubali powder are broken into 0.01-200 μ m fine powder, and all the other lamiophlomis rotatas, whin, turmeric and German tamarisk four flavor choppings add water, ethanol or vinegar and decocted 0.5-3 hour; Add above-mentioned fine powder again, stir, be placed to room temperature; Drying is taken out, and is ground into fine powder; According to conventional method, process any preparation;
Or the preparation method is for to be broken into 0.01-200 μ m fine powder with Chinese prickly ash, Cornu Bubali powder; All the other lamiophlomis rotatas, whin, turmeric and German tamarisk four flavor choppings add water, ethanol or vinegar and decocted 0.5-3 hour, add above-mentioned fine powder again; Stir; Put into the aluminium dish after reducing to room temperature, go into the vacuum refrigeration storehouse, be cooled to-15 ℃～-40 ℃.Begin to vacuumize, the taking-up that opens a position after 10-24 hour is pulverized, and processes said formulation on any pharmacy by conventional method.
The present invention provides the above-mentioned analgesic plaster preparation of being processed by lamiophlomis rotata 5-15 part, whin 2-10 part, turmeric 15-40 part, Chinese prickly ash 7-20 part, cornu bubali 5-15 part and German tamarisk 15-40 part bulk drug or the standard finger-print detection method of extract; Wherein chromatographic condition is: with octadecane base key and silica gel is filling agent; (3-9: 70-90) be the A phase, acetonitrile is a B phase composition moving phase with tetrahydrofuran-0.5% phosphoric acid; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, and the A phase: 80～95%, the B phase 20～5%, 60～100min, the A phase: 55～85%, the B phase 45～15%, 100～160min, the A phase: 30～77%, the B phase 70～23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures, sample size 1～25ul;
The volume ratio of said acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 40～10: 4.8～7.2: 55.2～82.8;
Said chromatographic condition is preferably: with octadecane base key and silica gel is filling agent, is the A phase with 6.4: 73.6 tetrahydrofurans-0.5% phosphoric acid, and acetonitrile is a B phase composition moving phase, adopts the gradient elution mode; Its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak;
Said standard finger-print common characteristic fingerprint peaks has 18, and its peak area comprehensively accounts for more than 90% of total peak area; No. 10 chromatographic peaks are the characteristic peak of cyanidenon, and the relative retention time of the characteristic peak of cyanidenon and relative peak area are 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster;
The common characteristic peak is in the resulting analgesic plaster standard finger-print:
No. 1 peak: relative retention time 0.11～1.20 keeps area 0.18～11.62 relatively
No. 2 peaks: relative retention time 0.14～1.26 keeps area 0.42～27.39 relatively
No. 3 peaks: relative retention time 0.16～1.31 keeps area 0.26～16.97 relatively
No. 4 peaks: relative retention time 0.23～1.38 keeps area 0.16～10.43 relatively
No. 5 peaks: relative retention time 0.35～1.54 keeps area 0.56～36.44 relatively
No. 6 peaks: relative retention time 0.42～1.67 keeps area 5.71～79.24 relatively
No. 7 peaks: relative retention time 0.65～1.82 keeps area 0.48～31.27 relatively
No. 8 peaks: relative retention time 0.67～1.87 keeps area 0.36～23.47 relatively
No. 9 peaks: relative retention time 0.96～2.13 keeps area 0.53～34.45 relatively
No. 10 peaks: (reference peak: the cyanidenon peak) relative retention time 1, keeps area 1 relatively
No. 11 peaks: relative retention time 1.07～2.68 keeps area 2.01～83.28 relatively
No. 12 peaks: relative retention time 1.23～2.86 keeps area 0.41～26.65 relatively
No. 13 peaks: relative retention time 1.29～2.94 keeps area 1.47～72.10 relatively
No. 14 peaks: relative retention time 1.36～3.02 keeps area 0.15～9.75 relatively
No. 15 peaks: relative retention time 1.41～3.14 keeps area 0.84～54.61 relatively
No. 16 peaks: relative retention time 1.50～3.53 keeps area 1.63～71.33 relatively
No. 17 peaks: relative retention time 1.68～3.64 keeps area 0.35～22.71 relatively
No. 18 peaks: relative retention time 1.72～4.00 keeps area 2.82～127.89 relatively.
Preferably, be 1 with the relative retention time and the relative peak area of the chromatographic peak (No. 10 peaks) of cyanidenon, the common characteristic peak is in the resulting analgesic plaster standard finger-print:
No. 1 peak: relative retention time 0.11 keeps area 0.18～11.62 relatively
No. 2 peaks: relative retention time 0.14 keeps area 0.42～27.39 relatively
No. 3 peaks: relative retention time 0.16 keeps area 0.26～16.97 relatively
No. 4 peaks: relative retention time 0.23 keeps area 0.16～10.43 relatively
No. 5 peaks: relative retention time 0.35 keeps area 0.56～36.44 relatively
No. 6 peaks: relative retention time 0.42 keeps area 5.71～79.24 relatively
No. 7 peaks: relative retention time 0.65 keeps area 0.48～31.27 relatively
No. 8 peaks: relative retention time 0.67 keeps area 0.36～23.47 relatively
No. 9 peaks: relative retention time 0.96 keeps area 0.53～34.45 relatively
No. 10 peaks: (reference peak: the cyanidenon peak) relative retention time 1, keeps area 1 relatively
No. 11 peaks: relative retention time 1.07 keeps area 2.01～83.28 relatively
No. 12 peaks: relative retention time 1.23 keeps area 0.41～26.65 relatively
No. 13 peaks: relative retention time 1.29 keeps area 1.47～72.10 relatively
No. 14 peaks: relative retention time 1.36 keeps area 0.15～9.75 relatively
No. 15 peaks: relative retention time 1.41 keeps area 0.84～54.61 relatively
No. 16 peaks: relative retention time 1.50 keeps area 1.63～71.33 relatively
No. 17 peaks: relative retention time 1.68 keeps area 0.35～22.71 relatively
No. 18 peaks: relative retention time 1.72 keeps area 2.82～127.89 relatively.
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol or ethanol or acetonitrile and process the reference substance solution that concentration is 5～40ug/ml, the chromatographic condition identical with need testing solution measured, and obtains the retention time of cyanidenon peak characteristic peak;
The preparation method of said need testing solution is:
Cancellation pain emplastrum preparation 0.5～2g (the analgesic plaster liquid preparation is pressed 1: 10～1: 30 volume ratio), the methanol hydrochloride solution of adding 0.5～5mol/L, reflux 10～120 minutes; Filter; Precision is measured subsequent filtrate, with methyl alcohol or acetonitrile dissolving, as need testing solution;
Methanol hydrochloride solution is by any one replacement in the phosphoric acid acetonitrile solution of the hydrochloric acid acetonitrile solution of the phosphoric acid methanol solution of 0.5～5mol/L or 0.5～5mol/L or 0.5～5mol/L among the preparation method of said need testing solution;
Among the preparation method of said need testing solution, reflux is by ultrasonic Extraction 5～60 minutes or refluxing extraction 1～12 hour or any one replacement of 0.1～4 hour of microwave abstracting.
Analgesic plaster bulk drug composition of the present invention is preferably:
9.1 parts of lamiophlomis rotatas, 4.5 parts of whins, 40.9 parts in turmeric, 13.6 parts in Chinese prickly ash, 9.1 parts of cornu bubalis, 22.7 parts of German tamarisks;
The analgesic plaster bulk drug is formed or is preferably:
7.1 parts of lamiophlomis rotatas, 7.5 parts of whins, 20 parts in turmeric, 15.6 parts in Chinese prickly ash, 7.1 parts of cornu bubalis, 32.7 parts of German tamarisks;
The analgesic plaster bulk drug is formed or is preferably:
11.1 parts of lamiophlomis rotatas, 3.5 parts of whins, 50.9 parts in turmeric, 13.6 parts in Chinese prickly ash, 12 parts of cornu bubalis, 17 parts of German tamarisks.
Analgesic plaster can be the various emplastrums (the disclosed formulation of ZL200510080696.4) according to the prescription preparation of above-mentioned ZL200510080696.4 patent in the said test sample; Also can be according to the extract of the prescription of above-mentioned ZL200510080696.4 patent preparation or raw material directly preparation the Chinese medicine fine powder or prepare various oral clinical formulations according to the prescription of above-mentioned ZL200510080696.4 patent, like tablet, capsule, particle, sustained release preparation etc.
The present invention has that method is easy, stable, precision is high, favorable reproducibility, the characteristics that are easy to grasp.The method for building up of experimental example 1 cyanidenon collection of illustrative plates finger-print
1 instrument and reagent
1.1 instrument
Waters 2695 high performance liquid chromatographs, DAD (PDAD), EMPOWER TM 2 operating systems.
1.2 reagent
Tetrahydrofuran, chromatographically pure; Acetonitrile, chromatographically pure; Phosphoric acid is analyzed pure; Analgesic plaster is that XiZang QiZheng Tibetan pharmaceuticals Co., Ltd produces.
2 methods and result
2.1 chromatographic condition
With octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6), and acetonitrile is a B phase composition moving phase, adopts the gradient elution mode; Its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
2.2 the mensuration of characteristic fingerprint pattern
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 15ug/ml, according to 2005 editions or 2010 editions appendix high performance liquid chromatography tests of Chinese Pharmacopoeia, the chromatographic condition identical with need testing solution measured; Obtain cyanidenon peak retention time, obtain cyanidenon collection of illustrative plates II.
The method for building up of the standard finger-print I of experimental example 2 analgesic plasters
1. instrument and reagent
1.1 instrument
Waters 2695 high performance liquid chromatographs, DAD (PDAD), EMPOWER TM 2 operating systems.
1.2 reagent
Tetrahydrofuran, chromatographically pure; Acetonitrile, chromatographically pure; Phosphoric acid is analyzed pure; Analgesic plaster is that XiZang QiZheng Tibetan pharmaceuticals Co., Ltd produces (pressing ZL200510080696.4 embodiment 1).
2. method and result
2.1 chromatographic condition
With octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6), and acetonitrile is a B phase composition moving phase, adopts the gradient elution mode; Its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
2.2 the mensuration of finger-print
2.2.1 analgesic plaster finger-print preparation
The preparation method of need testing solution: cancellation pain emplastrum sample 2g, precision is weighed, the accurate methanol hydrochloride solution that adds 2.5mol/L, reflux 30 minutes filters, precision is measured subsequent filtrate 2ml, put in the 5ml measuring bottle, with methyl alcohol to scale, as need testing solution.
Need testing solution 10ul is injected high performance liquid chromatograph; Compare with cyanidenon peak retention time; No. 10 chromatographic peaks are the characteristic peak of cyanidenon; The relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) are 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print I of analgesic plaster.
2.2.2 confirming of total peak
Through 10 batches of analgesic plaster medicinal powder determining fingerprint patterns; Compare its chromatogram; Confirm 18 at total peak; Be object of reference with No. 10 fingerprint peakses (characteristic peak of cyanidenon) wherein, its peak area surpasses 10% of total peak area, and the 11 lot sample article of enumerating have that peak relative retention time and relative peak area statistics see the following form 1, table 2.
The relative retention time statistics at 11 batches of total peaks of table 1 analgesic plaster
The relative area statistics at 11 batches of total peaks of table 2 analgesic plaster
Description of drawings
Fig. 1 is the standard finger-print I of analgesic plaster
Fig. 2 is cyanidenon collection of illustrative plates II
Below further specify the present invention through specific embodiment, but be not construed as limiting the invention.
Embodiment
Embodiment 1:
1, analgesic plaster preparation:
Lamiophlomis rotata 9.1g, whin 4.5g, turmeric 40.9g, Chinese prickly ash 13.6g, cornu bubali 9.1g, German tamarisk 22.7g; Chinese prickly ash, Cornu Bubali powder are broken into the fine powder of 50 μ m.Four flavor choppings such as all the other lamiophlomis rotatas, boiling 0.5 hour adds above-mentioned fine powder again, stirs, and is placed to room temperature, and drying is taken out, and is ground into fine powder.Fine powder is packed in the non-woven bag, seal, stick at and process emplastrum on the adhesive plaster, packing promptly gets.
2, the analgesic plaster finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum sample 2g, precision is weighed, the accurate methanol hydrochloride solution that adds 2.5mol/L, reflux 30 minutes filters, precision is measured subsequent filtrate 2ml, put in the 5ml measuring bottle, with methyl alcohol to scale, as need testing solution.
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 15ug/ml; According to Chinese Pharmacopoeia 2005 editions or 2010 editions appendix high performance liquid chromatography tests, the chromatographic condition identical with need testing solution measured, and obtains cyanidenon peak retention time;
Chromatographic condition: with octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6), and acetonitrile is a B phase composition moving phase, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 30: 5: 72; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Embodiment 2:
1, analgesic plaster extract preparation:
Lamiophlomis rotata 9.1g, whin 4.5g, turmeric 40.9g, Chinese prickly ash 13.6g, cornu bubali 9.1g, German tamarisk 22.7g; Chinese prickly ash, Cornu Bubali powder are broken into the fine powder of 50 μ m.Four flavor choppings such as all the other lamiophlomis rotatas, boiling 0.5 hour adds above-mentioned fine powder again, stirs, and is placed to room temperature, and drying is taken out, and is ground into fine powder, promptly gets extract.
2, the analgesic plaster finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum extract sample 1.8g, precision is weighed, the accurate methanol hydrochloride solution that adds 3.5mol/L; Reflux 40 minutes or, filter, precision is measured subsequent filtrate 2ml; Put in the 5ml measuring bottle, with methyl alcohol to scale, as need testing solution.
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 15ug/ml; According to Chinese Pharmacopoeia 2005 editions or 2010 editions appendix high performance liquid chromatography tests, the chromatographic condition identical with need testing solution measured, and obtains cyanidenon peak retention time;
Chromatographic condition: with octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (5.4: 75.6), and acetonitrile is a B phase composition moving phase, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 20: 6.2: 60; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 1ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Embodiment 3:
1, the ointment for preparing according to ZL200510080696.4 embodiment 2
2, the analgesic plaster finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum ointment 20g, precision is weighed, the accurate methanol hydrochloride solution that adds 2.5mol/L, ultrasonic Extraction 30 minutes filters, precision is measured subsequent filtrate 2ml, put in the 5ml measuring bottle, with methyl alcohol to scale, as need testing solution;
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 15ug/ml; According to Chinese Pharmacopoeia 2005 editions or 2010 editions appendix high performance liquid chromatography tests, the chromatographic condition identical with need testing solution measured, and obtains cyanidenon peak retention time;
Chromatographic condition: with octadecane base key and silica gel is filling agent; With tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6) is the A phase; Acetonitrile is a B phase composition moving phase, adopts the gradient elution mode, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 20: 7.2: 80; Its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Embodiment 4:
1, the Babu plaster for preparing according to ZL200510080696.4 embodiment 3
2, finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum Babu plaster 10g, precision is weighed, the accurate methanol hydrochloride solution that adds 3.5mol/L; Microwave abstracting 2 hours filters, and precision is measured subsequent filtrate 2ml; Put in the 5ml measuring bottle, with methyl alcohol to scale, as need testing solution.
Chromatographic condition: with octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (4.4: 80.6), and acetonitrile is a B phase composition moving phase, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 15: 5.2: 60; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 20%, 100～160min 80%,, the A phase: 70%, the B phase 30%; The detection wavelength is 350 ± 2nm, flow velocity 1.2ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Embodiment 5:
1, analgesic plaster preparation:
Lamiophlomis rotata 9.1g, whin 4.5g, turmeric 40.9g, Chinese prickly ash 13.6g, cornu bubali 9.1g, German tamarisk 22.7g; Chinese prickly ash, Cornu Bubali powder are broken into the fine powder of 50 μ m.Four flavor choppings such as all the other lamiophlomis rotatas, boiling 0.5 hour adds above-mentioned fine powder again, stirs, and is placed to room temperature, and drying is taken out, and is ground into fine powder.Fine powder is packed in the non-woven bag, seal, stick at and process emplastrum on the adhesive plaster, packing promptly gets.
2, the analgesic plaster finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum sample 2g, precision is weighed, the accurate methanol hydrochloride solution that adds 2.5mol/L; Ultrasonic Extraction 30 minutes filters, and precision is measured subsequent filtrate 2ml; Put in the 5ml measuring bottle, be dissolved to scale, as need testing solution with acetonitrile.
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 20ug/ml; According to Chinese Pharmacopoeia 2005 editions or 2010 editions appendix high performance liquid chromatography tests, the chromatographic condition identical with need testing solution measured, and obtains cyanidenon peak retention time;
Chromatographic condition: with octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6), and acetonitrile is a B phase composition moving phase, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 20: 5.2: 65.2; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Embodiment 6:
1, analgesic plaster preparation:
Lamiophlomis rotata 9.1g, whin 4.5g, turmeric 40.9g, Chinese prickly ash 13.6g, cornu bubali 9.1g, German tamarisk 22.7g; Chinese prickly ash, Cornu Bubali powder are broken into the fine powder of 50 μ m.Four flavor choppings such as all the other lamiophlomis rotatas, boiling 0.5 hour adds above-mentioned fine powder again, stirs, and is placed to room temperature, and drying is taken out, and is ground into fine powder.Fine powder is packed in the non-woven bag, seal, stick at and process emplastrum on the adhesive plaster, packing promptly gets.
2, the analgesic plaster finger-print detects
The preparation method of need testing solution: cancellation pain emplastrum sample 2g, precision is weighed, the accurate phosphoric acid acetonitrile solution that adds 2.5mol/L; Microwave abstracting 2 hours filters, and precision is measured subsequent filtrate 2ml; Put in the 5ml measuring bottle, be dissolved to scale, as need testing solution with acetonitrile.
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol and process the reference substance solution that concentration is 18ug/ml; According to Chinese Pharmacopoeia 2005 editions or 2010 editions appendix high performance liquid chromatography tests, the chromatographic condition identical with need testing solution measured, and obtains cyanidenon peak retention time
Chromatographic condition: with octadecane base key and silica gel is filling agent, is the A phase with tetrahydrofuran-0.5% phosphoric acid (6.4: 73.6), and acetonitrile is a B phase composition moving phase, and the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 20: 5.2: 65.2; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
Need testing solution 10ul is injected high performance liquid chromatograph, with the relative retention time and the relative peak area at No. 10 peaks (characteristic peak of cyanidenon) be 1, calculate the relative retention time and the relative peak area at other peaks, promptly obtain the standard finger-print of analgesic plaster.
Claims (9)
1. analgesic plaster preparation finger detection method is characterized in that this method is:
Cancellation pain emplastrum preparation 0.5～2g, the methanol hydrochloride solution of adding 0.5～5mol/L, reflux 10～120 minutes filters, and precision is measured subsequent filtrate, with methyl alcohol or acetonitrile dissolving, as need testing solution;
Wherein chromatographic condition is: with octadecane base key and silica gel is filling agent, and with 3-9: 70-90 tetrahydrofuran-0.5% phosphoric acid is the A phase, and acetonitrile is a B phase composition moving phase; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, and the A phase: 80～95%, the B phase 20～5%, 60～100min, the A phase: 55～85%, the B phase 45～15%, 100～160min, the A phase: 30～77%, the B phase 70～23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures, sample size 1～25ul;
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol or ethanol or acetonitrile and process the reference substance solution that concentration is 5～40ug/ml, the chromatographic condition identical with need testing solution measured, and obtains the retention time of cyanidenon peak characteristic peak;
Determination method: need testing solution is injected high performance liquid chromatograph promptly get;
Said analgesic plaster preparation is processed by following bulk drugs: lamiophlomis rotata 5-15 part, whin 2-10 part, turmeric 15-60 part, Chinese prickly ash 7-20 part, cornu bubali 5-15 part and German tamarisk 15-40 part.
2. analgesic plaster preparation finger detection method is characterized in that this method is:
Cancellation pain emplastrum liquid preparation is by the methanol hydrochloride solution of volume ratio adding 0.5～5mol/L of 1: 10～1: 30, and reflux 10～120 minutes filters, and precision is measured subsequent filtrate, with methyl alcohol or acetonitrile dissolving, as need testing solution;
Wherein chromatographic condition is: with octadecane base key and silica gel is filling agent, and with 3-9: 70-90 tetrahydrofuran-0.5% phosphoric acid is the A phase, and acetonitrile is a B phase composition moving phase; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, and the A phase: 80～95%, the B phase 20～5%, 60～100min, the A phase: 55～85%, the B phase 45～15%, 100～160min, the A phase: 30～77%, the B phase 70～23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures, sample size 1～25ul;
The preparation of reference substance solution: it is an amount of to get the cyanidenon reference substance; Precision is weighed; Add methyl alcohol or ethanol or acetonitrile and process the reference substance solution that concentration is 5～40ug/ml, the chromatographic condition identical with need testing solution measured, and obtains the retention time of cyanidenon peak characteristic peak;
Determination method: need testing solution is injected high performance liquid chromatograph promptly get;
Said analgesic plaster preparation is processed by following bulk drugs: lamiophlomis rotata 5-15 part, whin 2-10 part, turmeric 15-60 part, Chinese prickly ash 7-20 part, cornu bubali 5-15 part and German tamarisk 15-40 part.
3. like claim 1 or 2 arbitrary described fingerprint atlas detection methods, it is characterized in that the volume ratio of acetonitrile, tetrahydrofuran, 0.5% phosphoric acid is: 40～10: 4.8～7.2: 55.2～82.8.
4. fingerprint atlas detection method as claimed in claim 3 is characterised in that wherein chromatographic condition is: with octadecane base key and silica gel is filling agent, is the A phase with 6.4: 73.6 tetrahydrofuran-0.5% phosphoric acid, and acetonitrile is a B phase composition moving phase; Adopt the gradient elution mode, its elution time and moving phase ratio are: 0～60min, A phase: B phase 5%, 60～100min 95%; The A phase: B phase 15%, 100～160min 85%,, the A phase: 77%, the B phase 23%; The detection wavelength is 350 ± 2nm, flow velocity 0.5～1.5ml/min, 20～50 ℃ of column temperatures; Sample size 10ul, theoretical cam curve should be not less than 4000 by the cyanidenon peak.
5. like claim 1 or 2 arbitrary described fingerprint atlas detection methods, be characterised in that finger-print common characteristic fingerprint peaks has 18, its peak area comprehensively accounts for more than 90% of total peak area; No. 10 chromatographic peaks are the characteristic peak of cyanidenon, and the relative retention time of the characteristic peak of cyanidenon and relative peak area are 1, calculate the relative retention time and the relative peak area at other peaks, and the standard finger-print common characteristic peak that promptly obtains analgesic plaster is:
No. 1 peak: relative retention time 0.11～1.20 keeps area 0.18～11.62 relatively
No. 2 peaks: relative retention time 0.14～1.26 keeps area 0.42～27.39 relatively
No. 3 peaks: relative retention time 0.16～1.31 keeps area 0.26～16.97 relatively
No. 4 peaks: relative retention time 0.23～1.38 keeps area 0.16～10.43 relatively
No. 5 peaks: relative retention time 0.35～1.54 keeps area 0.56～36.44 relatively
No. 6 peaks: relative retention time 0.42～1.67 keeps area 5.71～79.24 relatively
No. 7 peaks: relative retention time 0.65～1.82 keeps area 0.48～31.27 relatively
No. 8 peaks: relative retention time 0.67～1.87 keeps area 0.36～23.47 relatively
No. 9 peaks: relative retention time 0.96～2.13 keeps area 0.53～34.45 relatively
No. 10 peaks: relative retention time 1 keeps area 1 relatively
No. 11 peaks: relative retention time 1.07～2.68 keeps area 2.01～83.28 relatively
No. 12 peaks: relative retention time 1.23～2.86 keeps area 0.41～26.65 relatively
No. 13 peaks: relative retention time 1.29～2.94 keeps area 1.47～72.10 relatively
No. 14 peaks: relative retention time 1.36～3.02 keeps area 0.15～9.75 relatively
No. 15 peaks: relative retention time 1.41～3.14 keeps area 0.84～54.61 relatively
No. 16 peaks: relative retention time 1.50～3.53 keeps area 1.63～71.33 relatively
No. 17 peaks: relative retention time 1.68～3.64 keeps area 0.35～22.71 relatively
No. 18 peaks: relative retention time 1.72～4.00 keeps area 2.82～127.89 relatively.
6. fingerprint atlas detection method as claimed in claim 5 is characterised in that the common characteristic peak is in the analgesic plaster preparation finger:
No. 1 peak: relative retention time 0.11 keeps area 0.18～11.62 relatively
No. 2 peaks: relative retention time 0.14 keeps area 0.42～27.39 relatively
No. 3 peaks: relative retention time 0.16 keeps area 0.26～16.97 relatively
No. 4 peaks: relative retention time 0.23 keeps area 0.16～10.43 relatively
No. 5 peaks: relative retention time 0.35 keeps area 0.56～36.44 relatively
No. 6 peaks: relative retention time 0.42 keeps area 5.71～79.24 relatively
No. 7 peaks: relative retention time 0.65 keeps area 0.48～31.27 relatively
No. 8 peaks: relative retention time 0.67 keeps area 0.36～23.47 relatively
No. 9 peaks: relative retention time 0.96 keeps area 0.53～34.45 relatively
No. 10 peaks: relative retention time 1 keeps area 1 relatively
No. 11 peaks: relative retention time 1.07 keeps area 2.01～83.28 relatively
No. 12 peaks: relative retention time 1.23 keeps area 0.41～26.65 relatively
No. 13 peaks: relative retention time 1.29 keeps area 1.47～72.10 relatively
No. 14 peaks: relative retention time 1.36 keeps area 0.15～9.75 relatively
No. 15 peaks: relative retention time 1.41 keeps area 0.84～54.61 relatively
No. 16 peaks: relative retention time 1.50 keeps area 1.63～71.33 relatively
No. 17 peaks: relative retention time 1.68 keeps area 0.35～22.71 relatively
No. 18 peaks: relative retention time 1.72 keeps area 2.82～127.89 relatively.
7. like claim 1,2,4 or 6 any one described fingerprint atlas detection methods, it is characterized in that methanol hydrochloride solution among the preparation method of need testing solution of sample introduction is by any one replacement in the phosphoric acid acetonitrile solution of the hydrochloric acid acetonitrile solution of the phosphoric acid methanol solution of 0.5～5mol/L or 0.5～5mol/L or 0.5～5mol/L; Reflux is by ultrasonic Extraction 5～60 minutes or refluxing extraction 1～12 hour or any one replacement of 0.1～4 hour of microwave abstracting.
8. fingerprint atlas detection method as claimed in claim 3 is characterized in that methanol hydrochloride solution among the preparation method of need testing solution of sample introduction is by any one replacement in the phosphoric acid acetonitrile solution of the hydrochloric acid acetonitrile solution of the phosphoric acid methanol solution of 0.5～5mol/L or 0.5～5mol/L or 0.5～5mol/L; Reflux is by ultrasonic Extraction 5～60 minutes or refluxing extraction 1～12 hour or any one replacement of 0.1～4 hour of microwave abstracting.
9. fingerprint atlas detection method as claimed in claim 5 is characterized in that methanol hydrochloride solution among the preparation method of need testing solution of sample introduction is by any one replacement in the phosphoric acid acetonitrile solution of the hydrochloric acid acetonitrile solution of the phosphoric acid methanol solution of 0.5～5mo1/L or 0.5～5mol/L or 0.5～5mol/L; Reflux is by ultrasonic Extraction 5～60 minutes or refluxing extraction 1～12 hour or any one replacement of 0.1～4 hour of microwave abstracting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010126328XA CN101797360B (en) | 2010-03-15 | 2010-03-15 | Fingerprint detection method for analgesic plaster |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010126328XA CN101797360B (en) | 2010-03-15 | 2010-03-15 | Fingerprint detection method for analgesic plaster |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101797360A CN101797360A (en) | 2010-08-11 |
| CN101797360B true CN101797360B (en) | 2012-02-29 |
Family
ID=42593301
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010126328XA Active CN101797360B (en) | 2010-03-15 | 2010-03-15 | Fingerprint detection method for analgesic plaster |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101797360B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102058849B (en) * | 2010-12-17 | 2012-06-27 | 西藏奇正藏药股份有限公司 | Application of Tibetan medicine composite to preparing alcohol withdrawal medicines |
| CN102028918B (en) * | 2010-12-17 | 2012-01-04 | 西藏奇正藏药股份有限公司 | Application of Tibetan medicinal composition to preparation of medicament for treating cerebral circulation insufficiency |
| CN103638444B (en) * | 2012-12-20 | 2020-07-14 | 浙江农林大学 | Effective part of Qizheng pain-relieving plaster and its preparation method |
| CN104237440B (en) * | 2014-10-09 | 2015-10-21 | 成都中医药大学 | The HPLC detection method of lamiophlomis rotata and finger-print detection technique |
| CN107102070B (en) * | 2017-03-25 | 2019-11-08 | 甘肃奇正藏药有限公司 | Analgesic plaster finger print quality detecting method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709445A (en) * | 2005-07-06 | 2005-12-21 | 甘肃奇正藏药有限公司 | Tibetan medicine for major functions of acute-chronic sprain-contusion, rheumatism, rheumatoid diseases, and its preparing method |
| CN101446573A (en) * | 2008-12-29 | 2009-06-03 | 甘肃奇正藏药有限公司 | Method for measuring content of luteolin in lamiophlomis rotata pharmaceutical preparation by liquid chromatography |
-
2010
- 2010-03-15 CN CN201010126328XA patent/CN101797360B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709445A (en) * | 2005-07-06 | 2005-12-21 | 甘肃奇正藏药有限公司 | Tibetan medicine for major functions of acute-chronic sprain-contusion, rheumatism, rheumatoid diseases, and its preparing method |
| CN101446573A (en) * | 2008-12-29 | 2009-06-03 | 甘肃奇正藏药有限公司 | Method for measuring content of luteolin in lamiophlomis rotata pharmaceutical preparation by liquid chromatography |
Non-Patent Citations (2)
| Title |
|---|
| 刘兰生等.高效液相色谱法测定奇正消痛贴膏中木犀草素的含量.《中国药事》.2006,第20卷(第7期),第418－419页. * |
| 董福慧.奇正消痛贴膏治关节炎的临床疗效观察.《继续医学教育》.2005,第19卷(第7期),第77-81页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101797360A (en) | 2010-08-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101797360B (en) | Fingerprint detection method for analgesic plaster | |
| CN108226321B (en) | Fingerprint detection method and fingerprint of fructus piperis longi and fructus piperis longi stomach-ache granules | |
| CN104730171B (en) | Radix Paeoniae Rubra, Flos Lonicerae multi-target ingredient content assaying method in a kind of compound Chinese medicinal preparation | |
| CN101766771A (en) | Quality control method of medicine for gynecopathy treatment | |
| CN106822203B (en) | Radix angelicae pubescentis granules and preparation method and quality control method thereof | |
| CN109668970B (en) | Ultra-high performance liquid chromatography detection method for traditional Chinese medicine composition | |
| CN104459003A (en) | Construction method of standard finger-print and characteristic spectrum of Suoquan preparation and quality detection method | |
| CN101502549B (en) | Notoginsen triterpenes capsule as well as preparation method thereof and method for measuring content | |
| CN110441407A (en) | A kind of pool art tablet quality control method | |
| CN101647993A (en) | Medicament for treating flu and preparation and detection method thereof | |
| WO2024007538A1 (en) | Content measurement method for six alkaloid components in small meridian-activating pill | |
| CN101816753B (en) | Method for detecting quality of compound preparation for treating cold | |
| CN115266975B (en) | Method for measuring genistin content in chicken's gizzard-membrane and processed product decoction pieces, standard decoction and formula granules thereof | |
| CN112881540A (en) | Method for detecting bupleurum chinense and vinegar bupleurum chinense formula granules | |
| CN113759056B (en) | Characteristic spectrum of Chinese lobelia and preparation thereof and construction method thereof | |
| CN103604898B (en) | The benefit heart relaxes the fingerprint atlas detection method of preparation | |
| CN103575823A (en) | Detection method of 8 chemical components in Tangminling preparation | |
| CN105497156A (en) | Preparation method of fleeceflower root life-extending preparation | |
| CN110133160B (en) | High performance liquid chromatography method for detecting characteristic spectrums of cowherb seed medicinal materials, decoction pieces, standard decoction and formula granules | |
| CN102813699A (en) | Pharmaceutical composition having effects of heat clearing, detoxifying and anti-inflammation and preparation method thereof | |
| CN111562324A (en) | Method for detecting content of multiple index components of medicinal preparation | |
| CN101874869B (en) | Method for measuring loganin content of Chinese medicinal composition | |
| CN105467051A (en) | Dachuanxiong tablet whole-time multi-wavelength fusion fingerprint quality control method | |
| CN106323794B (en) | Method for detecting process stability of gallbladder warming tablets | |
| CN103076424B (en) | Oxytropis medical material and detection method of oxytropis medical material preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |