CN1475802A - Chinese medicine quality control method - Google Patents
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Abstract
A method for controlling the quality of Chinese-medicinal materials, capejasmine fruit, by its fingerprint features that the jaseminoidin is used as the reference of fingerprint standard, the methanol solution is used as the reference solution, the retaining time and peak area of the chromatographic peak (S peak) of said jaseminoidin are used to calculate the ratio of retaining time to peak area.
Description
Invention field
The present invention relates to a kind of traditional Chinese medicine quality control method, particularly the method for quality control of Chinese medicine cape jasmine.
Background technology
Science and technology development provides condition for the Chinese medicine basic research, provides technical support for improving constantly traditional Chinese medicine quality.Traditional Chinese medicine fingerprint is a notion of using for reference the medical jurisprudence fingerprint, comprises Chinese medicine dna fingerprinting, protein fingerprint pattern and chemical fingerprint etc.; Usually said finger-print is meant chemical fingerprint; Traditional Chinese medicine fingerprint refers to that Chinese medicine is after suitably handling, adopt certain analysis means, obtain to indicate the total peak collection of illustrative plates of this Chinese medicine feature, by holding the feature of Chinese medicine fingerprint, differentiate the quality of Chinese medicine effectively, guarantee that Chinese crude drug and Chinese patent drug quality are relatively stable.
The research of traditional Chinese medicine fingerprint and application, not only can solve the difficult problem that China's traditional Chinese medicine authenticity of products difficulty is debated, also for strengthening the systematization and the standardization of Chinese crude drug plantation and processing specificationization, the inherent composition research of Chinese medicine, accelerate the development of the modernization of Chinese medicine, realize the assurance that science is provided in line with international standards.
Summary of the invention
The object of the invention is the finger-print that open cape jasmine is new.
The present invention is achieved by the following technical solutions:
Cape jasmine medicinal materials fingerprint examination criteria:
The preparation of need testing solution: get the meal 0.5g that cape jasmine crosses 24 mesh sieves, the accurate title, decide, and puts in the 100ml triangular flask, add methyl alcohol 100ml ultrasonic Extraction 15-30min, filter, get subsequent filtrate 50ml, water bath method, residue is separated with about 5ml is water-soluble, the granular activated carbon post of last 5g, wash with water near colourless after, add 30ml 60-80% ethanol elution, the ethanol eluate evaporate to dryness, the residue dissolve with methanol, constant volume becomes 10ml, shakes up, use membrane filtration, promptly get need testing solution;
The preparation of reference substance solution: get the Gardenoside reference substance, make solution that 1mg contains 0.5mg product solution in contrast with methyl alcohol; Assay method: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000); Chromatographic condition and system suitability test detect wavelength 238nm, and the acetonitrile-water gradient elution sees Table 1; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatogram, promptly; Is 1 calculating relative retention time and peak area ratio with Cape jasmine in the retention time and the peak area of the chromatographic peak (S peak) of glycosides; Finger-print and technical parameter: adopt high performance liquid chromatography, detect 10 batches of cape jasmine medicinal materials, be 1 hour writing time; The demarcation of total fingerprint peaks, (relative retention time): 1, (0.400) 2, (0.458) 3, (0.511) 4, (0.588) 5, (0.618) 6, (0.666) 7, (0.722) 8, (0.800) S, (1.000) 9, (1.084) 10, (1.109) 11, (1.195) 12, (1.235) 13, (1.290) 14, (1.815) 15, (1.832) 16, (1.850) 17, (1.864) 18, (1.889) 19, (1.905) 20, (1.918) 21, (1.940) 22, (1.950) 23, (1.962) 24, (1.972) 25, (1.991) 26, (2.019); Total fingerprint peaks area ratio: except that the Gardenoside peak, the peak area at other peak accounts for the number percent of total peak area all less than 10%, so peak area ratio is not done requirement; The part peak shape is described: the 14-26 peak is not for reaching the group peak of baseline separation, and what have is broad peak; Non-total peak area: test sample collection of illustrative plates and reference fingerprint compare, and the non-total peak total area must not be greater than 10% of total peak area.
Cape jasmine semi-manufacture finger-print examination criteria: cape jasmine semi-manufacture preparation: get cape jasmine, be ground into meal, the 65-80% ethanol percolation of doubly measuring with 7-9, collect percolate, reclaim ethanol, be concentrated into the clear cream that 55-70 ℃ of relative density is 1.00-1.20, add 3-5 times of water gaging dilution, stir evenly, 0-4 ℃ refrigerates 45-50 hour, filters, 55-70 ℃ of relative density of filtrate decompression simmer down to is the clear cream of 1.00-1.20, last activated-charcoal column is used the distilled water wash-out earlier, and is colourless to eluent, doubly measure the 65-80% ethanol elution with 9-11 again, collect eluent, decompression recycling ethanol also concentrates, and gets the cape jasmine semi-manufacture; The preparation of need testing solution: get cape jasmine semi-manufacture 1ml, water bath method adds methanol solvate, and the dilution constant volume becomes 2ml, crosses 0.45 μ m filter membrane, makes need testing solution;
The preparation of Gardenoside reference substance solution: get the Gardenoside reference substance, make the solution that every 1ml contains 0.5mg with methyl alcohol and compare product solution;
Assay method: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000); Chromatographic condition and system suitability test: detect wavelength 238nm, the acetonitrile-water gradient elution sees Table 1; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatograms, promptly; With the retention time of the chromatographic peak (S peak) of Gardenoside and peak area is 1 to calculate relative retention time and peak area ratio; Finger-print and technical parameter thereof: the peak number (relative retention time) of total fingerprint peaks: 1 (0.404) 2 (0.463) 3 (0.511) 4 (0.589) 5 (0.618) 6 (0.667) 7 (0.723) 8 (0.800) S (1.000) 9 (1.084) 10 (1.110) 11 (1.195) 12 (1.236) 13 (1.275); Peak area ratio: 1: 7: S=(0.121-0.153): (0.124-0.137): (1.000); Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak total area must not be greater than 5% of peak area;
The used instrument reagent of the half-finished finger-print of above cape jasmine and cape jasmine:
High performance liquid chromatograph: Waters2695 pump; Waters 2996 UV-detector; The Millcnnium32 chromatographic work station; Diamonsil C18 post (5 μ m, 4.6mm * 150mm); Acetonitrile is chromatographically pure (U.S. J.T.Baker); The Wahaha pure water; Methyl alcohol is chromatographically pure (Beijing prosperousization Fine Chemical Works);
Table 1 eluent gradient wash-out table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
The present invention discloses cape jasmine and the half-finished finger-print of cape jasmine first, and this makes cape jasmine and the half-finished quality control of cape jasmine, and more science is accurate.
Embodiment 1The preparation of cape jasmine semi-manufacture
Get cape jasmine, be ground into meal, according to the percolation under the liquid extract item (" appendix IO of Chinese pharmacopoeia version in 2000),, collect percolate with 70% ethanol percolation of 8 times of amounts, reclaim ethanol, being concentrated into relative density is the clear cream of 1.10 (60 ℃ of mensuration), adds 4 times of water gaging dilutions, stirs evenly, 0-4 ℃ refrigerates 48 hours, filter, filtrate decompression simmer down to relative density is the clear cream of 1.10 (60 ℃ of mensuration), last activated-charcoal column, use earlier the distilled water wash-out, colourless to eluent, with 10 times of amount 70% ethanol elutions, collect eluent again, decompression recycling ethanol also concentrates, and gets the cape jasmine semi-manufacture.Cape jasmine semi-manufacture finger-print examination criteria: the preparation of need testing solution: get cape jasmine semi-manufacture 1ml, water bath method adds methanol solvate, and the dilution constant volume becomes 2ml, crosses 0.45 μ m filter membrane, makes need testing solution; The preparation of Gardenoside reference substance solution: get the Gardenoside reference substance, make the solution that every 1ml contains 0.5mg with methyl alcohol and compare product solution;
Assay method: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000); Chromatographic condition and system suitability test: detect wavelength 238nm, the acetonitrile-water gradient elution sees Table 1; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatograms, promptly; With the retention time of the chromatographic peak (S peak) of Gardenoside and peak area is 1 to calculate relative retention time and peak area ratio; Finger-print and technical parameter thereof: the peak number (relative retention time) of total fingerprint peaks: 1 (0.404) 2 (0.463) 3 (0.511) 4 (0.589) 5 (0.618) 6 (0.667) 7 (0.723) 8 (0.800) S (1.000) 9 (1.084) 10 (1.110) 11 (1.195) 12 (1.236) 13 (1.275); Peak area ratio: 1: 7: S=(0.121-0.153): (0.124-0.137): (1.000); Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak total area must not be greater than 5% of peak area; The used instrument reagent of the half-finished finger-print of cape jasmine: high performance liquid chromatograph: Waters2695 pump; Waters 2996 UV-detector; The Millcnnium32 chromatographic work station; Diamonsil C18 post (5 μ m, 4.6mm * 150mm); Acetonitrile is chromatographically pure (U.S. J.T.Baker); The Wahaha pure water; Methyl alcohol is chromatographically pure (Beijing prosperousization Fine Chemical Works);
Table 1 eluent gradient wash-out table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
Embodiment 2Cape jasmine medicinal materials fingerprint examination criteria
The preparation of need testing solution: get the meal 0.5g that cape jasmine crosses 24 mesh sieves, the accurate title, decide, and puts in the 100ml triangular flask, add methyl alcohol 100ml ultrasonic Extraction 20min, filter, get subsequent filtrate 50ml, water bath method, residue is separated with about 5ml is water-soluble, the granular activated carbon post of last 5g, wash with water near colourless after, add 30ml 70% ethanol elution, the ethanol eluate evaporate to dryness, the residue dissolve with methanol, constant volume becomes 10ml, shakes up, with 0.45 μ m membrane filtration, promptly get need testing solution;
The preparation of reference substance solution: get the Gardenoside reference substance, make solution that 1mg contains 0.5mg product solution in contrast with methyl alcohol; Assay method: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000); Chromatographic condition and system suitability test detect wavelength 238nm, and the acetonitrile-water gradient elution sees Table 1; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatogram, promptly; Is 1 calculating relative retention time and peak area ratio with Cape jasmine in the retention time and the peak area of the chromatographic peak (S peak) of glycosides; Finger-print and technical parameter: adopt high performance liquid chromatography, detect 10 batches of cape jasmine medicinal materials, be 1 hour writing time; The demarcation of total fingerprint peaks, (relative retention time): 1, (0.400) 2, (0.458) 3, (0.511) 4, (0.588) 5, (0.618) 6, (0.666) 7, (0.722) 8, (0.800) S, (1.000) 9, (1.084) 10, (1.109) 11, (1.195) 12, (1.235) 13, (1.290) 14, (1.815) 15, (1.832) 16, (1.850) 17, (1.864) 18, (1.889) 19, (1.905) 20, (1.918) 21, (1.940) 22, (1.950) 23, (1.962) 24, (1.972) 25, (1.991) 26, (2.019); Total fingerprint peaks area ratio: except that the Gardenoside peak, the peak area at other peak accounts for the number percent of total peak area all less than 10%, so peak area ratio is not done requirement; The part peak shape is described: the 14-26 peak is not for reaching the group peak of baseline separation, and what have is broad peak;
Non-total peak area: test sample collection of illustrative plates and reference fingerprint compare, and the non-total peak total area must not be greater than 10% of total peak area.
The used instrument reagent of cape jasmine finger-print:
High performance liquid chromatograph: Waters2695 pump; Waters 2996 UV-detector; The Millcnnium32 chromatographic work station; Diamonsil C18 post (5 μ m, 4.6mm * 150mm); Acetonitrile is chromatographically pure (U.S. J.T.Baker); The Wahaha pure water; Methyl alcohol is chromatographically pure (Beijing prosperousization Fine Chemical Works);
Table 1 eluent gradient wash-out table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
Claims (5)
1, the method for quality control of cape jasmine medicinal materials fingerprint detection, it is characterized in that this method is: the preparation of need testing solution: get the meal 0.5g that cape jasmine crosses 24 mesh sieves, the accurate title, decide, put in the 100ml triangular flask, add methyl alcohol 100ml ultrasonic Extraction 15-30min, filter, get subsequent filtrate 50ml, water bath method, residue is separated with about 5ml is water-soluble, the granular activated carbon post of last 5g, wash with water near colourless after, add the 30ml60-80% ethanol elution, ethanol eluate evaporate to dryness, residue dissolve with methanol, constant volume becomes 10ml, shake up, use membrane filtration, promptly get need testing solution; The preparation of reference substance solution: get the Gardenoside reference substance, make solution that 1mg contains 0.5mg product solution in contrast with methyl alcohol; Assay method: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000); Chromatographic condition and system suitability test detect wavelength 238nm, and the acetonitrile-water gradient elution sees Table; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatogram, promptly; Is 1 calculating relative retention time and peak area ratio with Cape jasmine in the retention time and the peak area of the chromatographic peak (S peak) of glycosides; Finger-print and technical parameter: adopt high performance liquid chromatography, detect 10 batches of cape jasmine medicinal materials, be 1 hour writing time; The demarcation of total fingerprint peaks, (relative retention time): 1, (0.400) 2, (0.458) 3, (0.511) 4, (0.588) 5, (0.618) 6, (0.666) 7, (0.722) 8, (0.800) S, (1.000) 9, (1.084) 10, (1.109) 11, (1.195) 12, (1.235) 13, (1.290) 14, (1.815) 15, (1.832) 16, (1.850) 17, (1.864) 18, (1.889) 19, (1.905) 20, (1.918) 21, (1.940) 22, (1.950) 23, (1.962) 24, (1.972) 25, (1.991) 26, (2.019); Total fingerprint peaks area ratio: except that the Gardenoside peak, the peak area at other peak accounts for the number percent of total peak area all less than 10%, so peak area ratio is not done requirement; The part peak shape is described: the 14-26 peak is not for reaching the group peak of baseline separation, and what have is broad peak; Non-total peak area: test sample collection of illustrative plates and reference fingerprint compare, and the non-total peak total area must not be greater than 10% of total peak area;
Surface low moves phase gradient wash-out table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
2; the method of quality control that cape jasmine medicinal materials fingerprint as claimed in claim 1 detects; it is characterized in that the preparation of need testing solution in this method: get the meal 0.5g that cape jasmine crosses 24 mesh sieves; the accurate title, decide; put in the 100ml triangular flask, add methyl alcohol 100ml ultrasonic Extraction 20min, filter; get subsequent filtrate 50ml; water bath method, residue is separated with about 5ml is water-soluble, the granular activated carbon post of last 5g; wash with water near colourless after; add 30ml 70% ethanol elution, ethanol eluate evaporate to dryness, residue dissolve with methanol; constant volume becomes 10ml; shake up,, promptly get need testing solution with 0.45 μ m membrane filtration.
3, a kind of cape jasmine semi-manufacture, it is characterized in that preparing with the following method: get cape jasmine, be ground into meal,, collect percolate with the 65-80% ethanol percolation that 7-9 doubly measures, reclaim ethanol, be concentrated into the clear cream that 55-70 ℃ of relative density is 1.00-1.20, add 3-5 times of water gaging dilution, stir evenly, 0-4 ℃ refrigerates 45-50 hour, filter, 55-70 ℃ of relative density of filtrate decompression simmer down to is the clear cream of 1.00-1.20, last activated-charcoal column, use earlier the distilled water wash-out, colourless to eluent, doubly measure the 65-80% ethanol elution with 9-11 again, collect eluent, decompression recycling ethanol also concentrates, and gets the cape jasmine semi-manufacture.
4, cape jasmine semi-manufacture as claimed in claim 3, it is characterized in that preparing with the following method: get cape jasmine, be ground into meal,, collect percolate with 70% ethanol percolation of 8 times of amounts, reclaim ethanol, be concentrated into 60 ℃ of following relative densities and be 1.10 clear cream, add 4 times of water gagings dilutions, stir evenly, 0-4 ℃ refrigerates 48 hours, filter, 60 ℃ of following relative densities of filtrate decompression simmer down to are 1.10 clear cream, last activated-charcoal column, use earlier the distilled water wash-out, colourless to eluent, with 10 times of amount 70% ethanol elutions, collect eluent again, decompression recycling ethanol also concentrates, and gets the cape jasmine semi-manufacture.
5, as the method for quality control of claim 3 or 4 described cape jasmine semi-manufacture finger-prints detections, it is characterized in that this method is: the preparation of need testing solution: get cape jasmine semi-manufacture 1ml, water bath method, add methanol solvate, the dilution constant volume becomes 2ml, crosses 0.45 μ m filter membrane, makes need testing solution; The preparation of Gardenoside reference substance solution: get the Gardenoside reference substance, make the solution that every 1ml contains 0.5mg with methyl alcohol and compare product solution; Assay method: according to high effective liquid chromatography for measuring; Chromatographic condition and system suitability test: detect wavelength 238nm, the acetonitrile-water gradient elution sees Table; Theoretical cam curve is calculated, and should not hang down 30000; Setting the need testing solution sample size is 10 μ l, writes down 60 minutes chromatograms, promptly; With the retention time of the chromatographic peak (S peak) of Gardenoside and peak area is 1 to calculate relative retention time and peak area ratio; Finger-print and technical parameter thereof: the peak number (relative retention time) of total fingerprint peaks: 1 (0.404) 2 (0.463) 3 (0.5 11) 4 (0.589) 5 (0.6 1 8), 6 (0.667) 7 (0.723) 8 (0.800) S (1.000) 9 (1.084) 10 (1.110) 11 (1.195) 12 (1.236) 13 (1.275); Peak area ratio: 1: 7: S=(0.121-0.153): (0.124-0.137): (1.000); Non-total peak area: test sample collection of illustrative plates and reference fingerprint are relatively; The non-total peak total area must not be greater than 5% of peak area;
Surface low moves phase gradient wash-out table
Time (min) flow velocity (ml/min) water % acetonitrile %
0 0.8 95 5
8 0.8 92 8
21 0.8 88 12
35 0.8 88 12
40 0.8 70 30
45 0.8 40 60
50 0.8 0 100
60 0.8 0 100
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100419423C (en) * | 2004-10-15 | 2008-09-17 | 成都优他制药有限责任公司 | Method for controlling quality of simply single drug |
| CN101343297B (en) * | 2008-07-08 | 2011-03-23 | 陕西科技大学 | Method for extracting and purifying gardenoside with absorbent charcoal |
| CN102692473A (en) * | 2011-03-22 | 2012-09-26 | 河北以岭医药研究院有限公司 | Method for determining fingerprints of cape jasmine fruit medicinal materials |
| CN109164178A (en) * | 2018-08-23 | 2019-01-08 | 四川新绿色药业科技发展有限公司 | Quality testing and the discrimination method of a kind of cape jasmine and its processed product |
| CN110554124A (en) * | 2019-09-24 | 2019-12-10 | 哈尔滨珍宝制药有限公司 | Method for measuring fingerprint spectrum of gardenia formula particles |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| UA78194C2 (en) * | 2000-12-08 | 2007-03-15 | Council Scient Ind Res | Method for revealing and identification of active components in extracts by chromatographic fingerprinting of "fingerprints" and processor of program data processing chromatograms |
| JP4886933B2 (en) * | 2001-01-12 | 2012-02-29 | カウンセル オブ サイエンティフィック アンド インダストリアル リサーチ | A novel method for standardization of chromatographic fingerprints and single medicines and formulations |
| CN1141575C (en) * | 2001-06-29 | 2004-03-10 | 天津市金士力药物研究开发有限公司 | Red sage medicine fingerprint establishing method and standard fingerprint atlas |
| CN1403809A (en) * | 2001-09-04 | 2003-03-19 | 洪筱坤 | Establishment and application of Chinese medicine chaacteristic fingerprint spectrum |
| CN100412545C (en) * | 2002-12-23 | 2008-08-20 | 北京采瑞医药有限公司 | Method for controlling quality of compound red sage root preparation used for treating cardio-cerebral vascualr disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100419423C (en) * | 2004-10-15 | 2008-09-17 | 成都优他制药有限责任公司 | Method for controlling quality of simply single drug |
| CN101343297B (en) * | 2008-07-08 | 2011-03-23 | 陕西科技大学 | Method for extracting and purifying gardenoside with absorbent charcoal |
| CN102692473A (en) * | 2011-03-22 | 2012-09-26 | 河北以岭医药研究院有限公司 | Method for determining fingerprints of cape jasmine fruit medicinal materials |
| CN102692473B (en) * | 2011-03-22 | 2015-08-19 | 石家庄以岭药业股份有限公司 | A kind of assay method of Fructus Gardeniae finger-print |
| CN109164178A (en) * | 2018-08-23 | 2019-01-08 | 四川新绿色药业科技发展有限公司 | Quality testing and the discrimination method of a kind of cape jasmine and its processed product |
| CN109164178B (en) * | 2018-08-23 | 2021-07-27 | 四川新绿色药业科技发展有限公司 | Quality detection and identification method for gardenia and processed products thereof |
| CN110554124A (en) * | 2019-09-24 | 2019-12-10 | 哈尔滨珍宝制药有限公司 | Method for measuring fingerprint spectrum of gardenia formula particles |
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Owner name: BEIJING CHINESE MEDICINE COLLEGE; TIANJIN HAIKE M Free format text: FORMER OWNER: BEIJING CHINESE MEDICINE COLLEGE; XINHAI PHARMACEUTICAL MANAGE CO.LTD.,BEIJING KEYUAN; BEIJING TIANCHENGXINHAI MEDICINE CO., LTD. Effective date: 20080530 |
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