CN101477102A - Quality standard of south yew decoction pieces and its detection method - Google Patents
Quality standard of south yew decoction pieces and its detection method Download PDFInfo
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Abstract
The invention provides quality standards and a detection method of southern yew decoction slices. The detection standards comprise: segment-shaped character, Kelly to filemot identification powder, not more than 12.5 percent of moisture, not more than 8.0 percent of total ash content, not more than 1.0 percent of acidic insoluble ash, not less than 18.0 percent of water-soluble extract and content measuring and not less than 0.03 percent of 10-deacetyl baccatine III (C29H36O11) calculated according to dry product, and the like. The detection method has good repeatability of detection result and accurate data, and can better control the quality of southern yew.
Description
Technical field
The present invention relates to Chinese medicine, be specifically related to Chinese medicine quality standard of south yew decoction pieces and detection method thereof.
Background technology
Chinese yew is on the books in medical science ancient books such as Compendium of Material Medica as Chinese medicine, traditional medication: leaf be used as medicine anti-inflammatory, pain relieving, treatment mange; Seed is used for the treatment of dyspepsia, indigestion and roundworm disease; Bark treatment diabetes.Scientific research in recent years finds that the Chinese yew plant contains efficient anticancer active constituents such as taxol, and being used for gynecological cancer, prostate, upper gastrointestinal, lung cancer etc. has better therapeutic effect.The taxol composition is described as " last line of defense of late tumor ", and " over 15 years one of the most effective anticancer preparation ", curative effect is certain, and is safe in utilization, developed a lot of medicines.The traditional Chinese medical science thinks that Chinese yew has the effect of supplementing qi and nourishing yin, enriching blood and promoting secretion of body fluid, detoxicating and fighting cancer.The present invention relates to southerm yew is one of kind of Chinese yew, and it is the most extensive to distribute in China, main product in Fujian, ground such as Zhejiang, Jiangxi, Hunan, Hubei, Sichuan.Quality standard of south yew decoction pieces does not still have in national standard and provincial standard and records, but, quality standard and detection method thereof are to guarantee the key of prepared slices of Chinese crude drugs quality, south yew decoction pieces is as the active drug of treatment cancer, and studying its quality standard and detection method thereof is an important process.
Summary of the invention
Technical matters to be solved by this invention is to overcome above-mentioned weak point, in conjunction with the characteristic of southerm yew, and research and design southerm yew quality standard and detection method thereof.
The quality standard that the invention provides south yew decoction pieces is:
This product is the band leaf branch bar of taxaceae plant southerm yew Taxusmairei (lemee etlevl.) S.Y.HuexLiu cultivation product;
[proterties] this product section of being shape.It is carefully cylindrical that stem is, the about 1～4mm of diameter, and the tangent plane that has has tiny marrow, and surperficial yellow green is to tawny, and the spray tool is indulged rib; Leaf cuts off more; Complete person's bar shaped, yellow green be to tawny, long 1.5～3cm, wide 0.25～0.35cm, top is gradually narrow, anxious point of tip or point gradually, petiole is short, the full edge of blade, nearly keratin, above in the arteries and veins protuberance obviously.Matter is crisp, frangibility, and gas is little, bitter, puckery;
[discriminating] this product powder yellow green is to tawny, and pore is more.Mesophyll parenchyma cell similar round is arranged more loose.Square or the polygon of cork cell includes brown yellow.Square, the polygon of parenchyma cell class.Spiral tracheid sees that minority is a scalariform tracheid more, diameter 20～60 μ m;
[inspection] moisture must not cross 12.5%.
Total ash must not cross 8.0%.
Acid-insoluble ash must not cross 1.0%.
[extract] must not be less than 18.0% according to water-soluble extractives.
[assay] this product is pressed dry product and is calculated, and contains 10-and takes off acetyl baccatin III (C
29H
36O
11) must not be less than 0.03%.
[nature and flavor with return through] are little sweet, bitter, and are flat; Slightly poisonous.Return kidney, the heart channel of Hang-Shaoyin.
[function with cure mainly] dispersing swelling and dissipating binds, the diuresis of stimulating the menstrual flow; Be used for diseases such as tumour, ephrosis, rheumatoid arthritis.
[usage and consumption] 3～9g.
[attention] may cause nausea, vomiting, fash.
[storage] puts ventilation, drying, lucifuge place.
Another purpose of the present invention has provided the detection method of south yew decoction pieces, and this method comprises the following steps:
Raw material: this product is the band leaf branch bar of taxaceae plant southerm yew Taxus mairei (lemee etlevl.) S.Y.Hu ex Liu cultivation product;
[proterties] this product section of being shape.It is carefully cylindrical that stem is, the about 1～4mm of diameter, and the tangent plane that has has tiny marrow, and surperficial yellow green is to tawny, and the spray tool is indulged rib; Leaf cuts off more; Complete person's bar shaped, yellow green be to tawny, long 1.5～3cm, wide 0.25～0.35cm, top is gradually narrow, anxious point of tip or point gradually, petiole is short, the full edge of blade, nearly keratin, above in the arteries and veins protuberance obviously.Matter is crisp, frangibility, and gas is little, bitter, puckery;
[discriminating]
(1) this product powder yellow green is to tawny, and pore is more.Mesophyll parenchyma cell similar round is arranged more loose.Square or the polygon of cork cell includes brown yellow.Square, the polygon of parenchyma cell class.Spiral tracheid sees that minority is a scalariform tracheid more, diameter 20～60 μ m;
(2) get this product powder 1g, add methyl alcohol 50ml, sonicated (40～50 ℃ of temperature) 40 minutes, filter, 40～50 ℃ of recovered under reduced pressure of filtrate are to doing, and residue adds the mixed liquor 45ml dissolving of water-methenyl choloride (1:2V/V), divide and take off a layer methenyl choloride, 40～50 ℃ of recovered under reduced pressure are to doing, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Other gets 10-and takes off acetyl baccatin III reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw need testing solution 5 μ l, reference substance solution 2 μ l, putting respectively on same silica gel H thin layer plate, is developping agent with methenyl choloride-ethyl acetate-methyl alcohol (8:1.2:0.8V/V), launches, take out, dry, spray is with 10% vanillic aldehyde concentrated sulfuric acid solution, and it is clear to be heated to spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
[inspection] moisture must not cross 12.5% according to aquametry (an appendix IX of Chinese Pharmacopoeia version in 2005 H first method).
Total ash must not be crossed 8.0% (an appendix IX of Chinese Pharmacopoeia version in 2005 K).
Acid-insoluble ash must not be crossed 1.0% (an appendix IX of Chinese Pharmacopoeia version in 2005 K).
[extract] measured according to hot dipping (an appendix X of Chinese Pharmacopoeia version in 2005 A) under the water-soluble extractives determination method item, must not be less than 18.0%.
[assay] measured according to high performance liquid chromatogram determination method (appendix VID of Chinese Pharmacopoeia version in 2005).
Chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, and water is Mobile phase B, and according to the form below carries out gradient elution; The detection wavelength is 232nm.Number of theoretical plate takes off acetyl baccatin III peak in 10-should be not less than 3500.
The preparation of reference substance solution is taken at 40～45 ℃ of drying under reduced pressure 10-more than 12 hours, and to take off acetyl baccatin III reference substance an amount of, accurately claims surely, adds methyl alcohol and make the solution that every 1ml contains 0.1mg, promptly.
This product powder (cross No. three sieve) 1g is got in the preparation of need testing solution, accurately claims surely, puts in the tool plug conical flask, adds methyl alcohol 50ml and places and spend the night, and (40～50 ℃ of temperature 40 minutes, filter sonicated for power 300W, frequency 50kHz; Add methyl alcohol 20 again, sonicated 20 minutes, merging filtrate, evaporated under reduced pressure, residue are with the mixed liquor 45ml of water-methenyl choloride (1:2V/V), and the gradation dissolving also is transferred in the separating funnel, divide and take off a layer methenyl choloride, water layer adds methenyl choloride 20ml again, and jolting is extracted, merge methenyl choloride liquid, 40～50 ℃ of evaporated under reduced pressure, residue is with dissolve with methanol and be transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
This product is pressed dry product and is calculated, and contains 10-and takes off acetyl baccatin III (C
29H
36O
11) must not be less than 0.03%.
Raw material sources of the present invention determine that through on-the-spot investigation, laboratory qualification the source is southerm yew Taxus mairei (lemee et levl.) S.Y.Hu ex Liu.
The present invention has set up the quality standard and the detection method thereof of south yew decoction pieces, the inventive method testing result good reproducibility, and data are accurate.Use method of the present invention can control the quality of south yew decoction pieces preferably,, provide the southerm yew that meets quality standard raw material, help the safe and effective of medicine, bigger using value is arranged for further preparing the southerm yew anticarcinogen.
Embodiment
Embodiment 1 differentiates
(1) proterties: this test is a real and fake discrimination.Detect by an unaided eye.Check check through 10 batch samples, shape is all up to specification.
(2) differentiate:
Microscopical identification: this test is a real and fake discrimination.This product powder is crossed 65 mesh sieves, and a little puts picking and drip chloral hydrate on the microslide,, on spirit lamp, add heat penetrationization and drip chloral hydrate, covered.Putting microscopically observes.Check check 10 batch samples, all have tangible microscopical identification feature.
Thin layer is differentiated: this test is a real and fake discrimination.
The preparation of need testing solution: get this product powder 1g, add methyl alcohol 50ml, sonicated 40 minutes, 45 ℃ of temperature filter, 45 ℃ of recovered under reduced pressure of filtrate are to doing, residue adds the mixed liquor 45ml dissolving of water-methenyl choloride (1:2V/V), divides and takes off a layer methenyl choloride, and 45 ℃ of recovered under reduced pressure are to doing, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.
The preparation of reference substance solution: get 10-and take off acetyl baccatin III reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution.
Developping agent: methenyl choloride-ethyl acetate-methyl alcohol (8:1.2:0.8V/V)
Silica gel plate: silica gel H plate
Point sample amount: reference substance 2 μ l, test sample 5 μ l
Developer: 10% vanillic aldehyde concentrated sulfuric acid solution.
Method: 110 ℃ in prefabricated silica gel H plate drying box baking activation in 30 minutes, put in the exsiccator cold after, use the kapillary point sample, reference substance 2 μ l, test sample 5 μ l; Put into the expansion cylinder that is placed with developping agent, ascending development 8～15cm takes out, and dries; Spray is with 10% vanillic aldehyde concentrated sulfuric acid solution, and it is clear that 105 ℃ of drying boxes are heated to the spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.Check check 4 batch samples, the result is up to specification.
Embodiment 2
Detect:
(1) moisture: this test is quality restriction.Measure according to aquametry (appendix IXH first method of Chinese Pharmacopoeia version in 2005).Get test sample 2～5g, be tiled in the flat measuring cup that is dried to constant weight, thickness is no more than 5mm, and accurate the title decides; Open bottle cap and bottle cap was built in 5 hours, move in the exsiccator, cooled off 30 minutes, accurate claim surely, dry 1 hour of said temperature, weigh again,, calculate the water cut of test sample according to the weight that subtracts mistake 105 ℃ of dryings.Check 5 batch samples, mean value is 11.3.Data see the following form.
(2) total ash: this test is quality restriction.Measure according to (appendix IXK of Chinese Pharmacopoeia version in 2005) ash determination method.This product was pulverized sieve No. two, got in the crucible that 3～5g puts ignition to constant weight, claimed decide weight (accurately to 0.01g), and was slowly red-hot, noted avoiding burning, and during to complete charing, the temperature to 580 that raises gradually makes complete ashing and to constant weight.According to residue weight, calculate the content (%) of total ash in the test sample.Check 5 batch samples, mean value is 4.3.Data see the following form.
(3) acid-insoluble ash: this test is quality restriction.Measure according to (an appendix IX of Chinese Pharmacopoeia version in 2005 K) ash determination method.Get ash determination method gained ash content, the careful about 10ml of watery hydrochloric acid that adds in crucible, cover crucible with surface plate, put in the water-bath and heated 10 minutes, surface plate washes with hot water 5ml, and washing lotion is incorporated in the crucible, filters with ashless filter paper, residue in the crucible washes with water on filter paper, and wash do not show the chloride reaction to washing lotion till.Residue moves in the same crucible together with filter paper, drying, ignition to constant weight.According to residue weight, calculate the content (%) of acid-insoluble ash in the test sample.Check 5 batch samples, mean value is 0.5.Data see the following form
5 batch samples are checked the item number certificate
| Sequence number | Lot number | Moisture | Total ash | Acid-insoluble ash |
| 1 | 080901 | 12.0 | 2.8 | 0.3 |
| 2 | 080902 | 12.3 | 3.7 | 0.5 |
| 3 | 080903 | 8.4 | 5.0 | 1.3 |
| 4 | 080920 | 11.9 | 3.9 | 0.0 |
| 5 | 081024 | 12.4 | 6.3 | 0.6 |
| Mean value | 11.3 | 4.3 | 0.5 |
(4) extract: this test is quality restriction.Measure according to hot dipping under the water-soluble extractives determination method item (an appendix X of Chinese Pharmacopoeia version in 2005 A).Get the about 2g of test sample, the accurate title, decide, and puts in the conical flask of 250ml, precision adds water 50ml, and close plug claims to decide weight, after leaving standstill 1 hour, connect reflux condensing tube, be heated to and boil, and keep little and boiled 1 hour, put and take off conical flask after cold, close plug claims to decide weight again, and water is supplied the weight that subtracts mistake, shake up, filter with dry filter, precision is measured filtrate 25ml, puts in the evaporating dish that is dried to constant weight, behind evaporate to dryness in the water-bath, in 105 ℃ of dryings 3 hours, put in the exsiccator cooling 30 minutes, accurately rapidly claim to decide weight.Except as otherwise herein provided, easily dry product calculate water-soluble extractives in the test sample content (%).Check 5 batch samples, mean value is 25.6.Data see the following form.
(5) assay: chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica; With the acetonitrile is mobile phase A, and water is Mobile phase B, and according to the form below carries out gradient elution; The detection wavelength is 232nm.Number of theoretical plate takes off acetyl baccatin III peak in 10-should be not less than 3500.
The preparation of reference substance solution is taken at 12 hours 10-of 40 ℃ of drying under reduced pressure, and to take off acetyl baccatin III reference substance an amount of, accurately claims surely, adds methyl alcohol and make the solution that every 1ml contains 0.1mg, promptly.
This product powder (cross No. three sieve) 1g is got in the preparation of need testing solution, accurately claims surely, puts in the tool plug conical flask, adds methyl alcohol 50ml and places and spend the night, and (power 300W, frequency 50kHz, 40 minutes, filter sonicated by 45 ℃; Add methyl alcohol 20 again, sonicated 20 minutes, merging filtrate, evaporated under reduced pressure, residue are with the mixed liquor 45ml of water-methenyl choloride (1:2V/V), and the gradation dissolving also is transferred in the separating funnel, divide and take off a layer methenyl choloride, water layer adds methenyl choloride 20ml again, and jolting is extracted, merge methenyl choloride liquid, 45 ℃ of evaporated under reduced pressure, residue is with dissolve with methanol and be transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.Check 5 batch samples, mean value is 0.048.Data see the following form.
Extract, assay data
| Sequence number | Lot number | Extract % | Assay % |
| 1 | 080901 | 26.1 | 0.050 |
| 2 | 080902 | 30.5 | 0.053 |
| 3 | 080903 | 24.0 | 0.052 |
| 4 | 080920 | 26.1 | 0.046 |
| 5 | 081024 | 21.3 | 0.040 |
| Mean value | 25.6 | 0.048 |
Claims (3)
1, a kind of quality standard of south yew decoction pieces is characterized in that its quality standard is:
(1) this product is the band leaf branch bar of taxaceae plant southerm yew Taxus mairei lemee et levl.S.Y.Hu ex Liu cultivation product;
(2) proterties: this product section of being shape, stem are carefully cylindrical, the about 1～4mm of diameter, and the tangent plane that has has tiny marrow, and surperficial yellow green is to tawny, and the spray tool is indulged rib; Leaf cuts off more; Complete person's bar shaped, yellow green be to tawny, long 1.5～3cm, wide 0.25～0.35cm, top is gradually narrow, anxious point of tip or point gradually, petiole is short, the full edge of blade, nearly keratin, above in the arteries and veins protuberance obviously; Matter is crisp, frangibility, and gas is little, bitter, puckery;
(3) differentiate: this product powder yellow green is to tawny, and pore is more, and mesophyll parenchyma cell similar round is arranged more loose, square or the polygon of cork cell includes brown yellow, square, the polygon of parenchyma cell class, spiral tracheid sees that minority is a scalariform tracheid more, diameter 20～60 μ m;
(4) moisture: must not cross 12.5%;
(5) total ash: must not cross 8.0%;
(6) acid-insoluble ash: must not cross 1.0%;
(7) extract: must not be less than 18.0% according to water-soluble extractives;
(8) assay: this product is pressed dry product and is calculated, and contains 10-and takes off acetyl baccatin III C
29H
36O
11Must not be less than 0.03%;
[nature and flavor with return through] are little sweet, bitter, and are flat; Slightly poisonous, return kidney, the heart channel of Hang-Shaoyin;
[function with cure mainly] dispersing swelling and dissipating binds, the diuresis of stimulating the menstrual flow; Be used for tumour, ephrosis, rheumatoid arthrosis inflammation;
[usage and consumption] 3～9g;
[attention] may cause nausea, vomiting, fash;
[storage] puts ventilation, drying, lucifuge place.
2, a kind of detection method of south yew decoction pieces is characterized in that this method comprises the following steps:
(1) raw material: this product is the band leaf branch bar of taxaceae plant southerm yew Taxus mairei lemee etlevl.S.Y.Hu ex Liu cultivation product;
(2) proterties: this product section of being shape, stem are carefully cylindrical, the about 1～4mm of diameter, and the tangent plane that has has tiny marrow, and surperficial yellow green is to tawny, and the spray tool is indulged rib; Leaf cuts off more; Complete person's bar shaped, yellow green be to tawny, long 1.5～3cm, wide 0.25～0.35cm, top is gradually narrow, anxious point of tip or point gradually, petiole is short, the full edge of blade, nearly keratin, above in the arteries and veins protuberance obviously, matter is crisp, frangibility, gas is little, bitter, puckery;
(3) differentiate:
1. powder microscopical identification: this product powder yellow green is to tawny, and pore is more, and mesophyll parenchyma cell similar round is arranged more loose, square or the polygon of cork cell includes brown yellow, square, the polygon of parenchyma cell class, spiral tracheid sees that minority is a scalariform tracheid more, diameter 20～60 μ m;
2. thin layer discrimination method: get this product powder 1g, add methyl alcohol 50ml, sonicated 40 minutes, 40～50 ℃ of temperature, filter, 50 ℃ of following recovered under reduced pressure of filtrate are to doing, and residue adds the mixed liquor 45ml dissolving of water-methenyl choloride 1:2V/V, divides and takes off a layer methenyl choloride, 40～50 ℃ of temperature, recovered under reduced pressure is to doing, and residue adds methyl alcohol 1ml makes dissolving, as need testing solution;
Other gets 10-and takes off acetyl baccatin III reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, product solution in contrast, according to appendix VIB test of thin-layered chromatography Chinese Pharmacopoeia version in 2005, draw need testing solution 5 μ l, reference substance solution 2 μ 1 put respectively on same silica gel H thin layer plate, are developping agent with methenyl choloride-ethyl acetate-methyl alcohol 8:1.2:0.8V/V, launch, take out, dry, spray is with 10% vanillic aldehyde concentrated sulfuric acid solution, it is clear to be heated to the spot colour developing, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
(4) moisture:, must not cross 12.5% according to appendix IXH first method of aquametry Chinese Pharmacopoeia version in 2005;
(5) total ash: must not cross an appendix IX of 8.0% Chinese Pharmacopoeia version in 2005 K;
(6) acid-insoluble ash: must not cross an appendix IX of 1.0% Chinese Pharmacopoeia version in 2005 K;
(7) extract: measure according to an appendix X of hot dipping Chinese Pharmacopoeia version in 2005 A under the water-soluble extractives determination method item, must not be less than 18.0%;
(8) assay is measured according to an appendix VI of high performance liquid chromatogram determination method Chinese Pharmacopoeia version in 2005 D:
The preparation of reference substance solution is taken at 12 hours 10-of 40～45 ℃ of drying under reduced pressure, and to take off acetyl baccatin III reference substance an amount of, accurately claims surely, adds methyl alcohol and make the solution that every 1ml contains 0.1mg, promptly;
The preparation of need testing solution is got this product powder and is crossed sieve 1g No. three, and accurate the title decides, and puts in the tool plug conical flask, adds methyl alcohol 50ml placement and spends the night, and (40～50 ℃ of temperature filtered sonicated for power 300W, frequency 50kHz in 40 minutes; Add methyl alcohol 20 again, sonicated 20 minutes, merging filtrate, evaporated under reduced pressure, residue are with the mixed liquor 45ml of water-methenyl choloride 1:2V/V, and the gradation dissolving also is transferred in the separating funnel, divide and take off a layer methenyl choloride, water layer adds methenyl choloride 20ml again, and jolting is extracted, merge methenyl choloride liquid, 40～50 ℃ of evaporated under reduced pressure, residue is with dissolve with methanol and be transferred in the 10ml measuring bottle, adds methyl alcohol to scale, shake up, promptly;
Assay method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, that is,
This product is pressed dry product and is calculated, and contains 10-and takes off acetyl baccatin III (C
29H
36O
11) must not be less than 0.03%.
3, method according to claim 2, it is characterized in that the chromatographic condition that high performance liquid chromatogram is measured in described step (8) assay is: octadecylsilane chemically bonded silica is a filling agent; With acetonitrile and water is moving phase, acetonitrile: water is that 78-20:22-80V/V carries out gradient elution; Time 0-35 minute; The detection wavelength is 232nm, and number of theoretical plate takes off acetyl baccatin III peak in 10-should be not less than 3500.
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| CN105372370A (en) * | 2015-11-23 | 2016-03-02 | 江苏红豆杉药业有限公司 | Method for detecting 10-deacetylbaccatin III in Chinese yew branches and leaves |
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| CN106568909B (en) * | 2015-12-29 | 2019-05-10 | 广西壮族自治区民族医药研究院 | The method of quality control of careless dragon medicinal material |
| CN106596759A (en) * | 2016-12-05 | 2017-04-26 | 广州中大南沙科技创新产业园有限公司 | Taxus chinensis branch and leaf extractive and HPLC analysis method of preparation thereof |
| CN106596759B (en) * | 2016-12-05 | 2019-07-02 | 广州中大南沙科技创新产业园有限公司 | A kind of HPLC analysis method of Ramulus et folium taxi cuspidatae extract and its preparation |
| CN108982504A (en) * | 2018-08-14 | 2018-12-11 | 中山大学 | A kind of tomentose pummelo fruit quality of medicinal material detection method |
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