CN102183593A - Method for detecting mannitol content of cordyceps hawkesii - Google Patents
Method for detecting mannitol content of cordyceps hawkesii Download PDFInfo
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- CN102183593A CN102183593A CN2011100706783A CN201110070678A CN102183593A CN 102183593 A CN102183593 A CN 102183593A CN 2011100706783 A CN2011100706783 A CN 2011100706783A CN 201110070678 A CN201110070678 A CN 201110070678A CN 102183593 A CN102183593 A CN 102183593A
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- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 title claims abstract description 19
- 229930195725 Mannitol Natural products 0.000 title claims abstract description 19
- 235000010355 mannitol Nutrition 0.000 title claims abstract description 19
- 239000000594 mannitol Substances 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 title abstract description 14
- 241000422920 Cordyceps gunnii Species 0.000 title abstract 5
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 6
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 32
- 241000190633 Cordyceps Species 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 17
- 235000009508 confectionery Nutrition 0.000 claims description 17
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 16
- 239000013558 reference substance Substances 0.000 claims description 16
- 238000012360 testing method Methods 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 11
- 238000001514 detection method Methods 0.000 claims description 10
- 238000005303 weighing Methods 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 6
- SFUVLEGIZGPPNN-UHFFFAOYSA-N (2-pyridin-2-ylacetyl) 2-pyridin-2-ylacetate Chemical compound C=1C=CC=NC=1CC(=O)OC(=O)CC1=CC=CC=N1 SFUVLEGIZGPPNN-UHFFFAOYSA-N 0.000 claims description 5
- WFDIJRYMOXRFFG-UHFFFAOYSA-N acetic anhydride Substances CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 5
- 230000021736 acetylation Effects 0.000 claims description 5
- 238000006640 acetylation reaction Methods 0.000 claims description 5
- 238000012216 screening Methods 0.000 claims description 5
- 239000006200 vaporizer Substances 0.000 claims description 5
- 238000004817 gas chromatography Methods 0.000 abstract description 3
- 238000005259 measurement Methods 0.000 abstract 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 abstract 1
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 239000007789 gas Substances 0.000 abstract 1
- 239000001257 hydrogen Substances 0.000 abstract 1
- 239000012088 reference solution Substances 0.000 abstract 1
- 239000012488 sample solution Substances 0.000 abstract 1
- 230000008016 vaporization Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 12
- 229940079593 drug Drugs 0.000 description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000004737 colorimetric analysis Methods 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 3
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- OISVCGZHLKNMSJ-UHFFFAOYSA-N 2,6-dimethylpyridine Chemical class CC1=CC=CC(C)=N1 OISVCGZHLKNMSJ-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- DMVOXQPQNTYEKQ-UHFFFAOYSA-N biphenyl-4-amine Chemical compound C1=CC(N)=CC=C1C1=CC=CC=C1 DMVOXQPQNTYEKQ-UHFFFAOYSA-N 0.000 description 1
- 238000006356 dehydrogenation reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- FJVZDOGVDJCCCR-UHFFFAOYSA-M potassium periodate Chemical compound [K+].[O-]I(=O)(=O)=O FJVZDOGVDJCCCR-UHFFFAOYSA-M 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a method for detecting mannitol content of cordyceps hawkesii, and belongs to the technical field of Chinese medicinal component content measurement. The mannitol content of the cordyceps hawkesii is measured by using gas chromatography, and the efficient and accurate measurement of the mannitol content of the special cordyceps hawkesii in Shanxi is realized. The method comprises the following steps of: preparing reference solution; preparing sample solution; and performing measurement by adopting a gas chromatograph, wherein the chromatography conditions comprise that: the initial temperature of column temperature is 220 DEG C and kept for 1 minute, the temperature is raised to between 220 and 230 DEG C at the rate of 10 DEG C per minute, the raised temperature is kept for 26 minutes, the temperature of a vaporizing chamber and the temperature a hydrogen flame ionization detector are 300 DEG C, the flow rate of H2 is 34 to 40ml/min, the flow rate of air is 350 to 400ml/min, the flow rate of N2 is 30 to 35/min, and the split ratio is 50:1. The method is simple, convenient, quick and accurate, has good repeatability, can measure the mannitol content of the special cordyceps hawkesii in Shanxi more scientifically and accurately, and effectively improves the quality of a product.
Description
Technical field
The invention belongs to traditional Chinese medicine ingredients assay technical field, the detection method of mannitol content in particularly a kind of incense stick cordycep.
Background technology
The Chinese medicine standard modern is the modern important component part of traditional Chinese medicine, and the Chinese crude drug standard modern is the basis of Chinese medicine standard modern.Having significant chemical constitution in the main flavour of a drug of selection Chinese patent drug is index components, measures its content, is the main means of chemical determination Chinese patent drug content; The consistent Chinese patent drug content assaying method of content assaying method of the contained index components of Chinese crude drug that invention and the general drug standards (particularly " Chinese pharmacopoeia) are recorded is one of main flow direction of chemical determination Chinese patent drug content.
At present, the detection method to mannitol content in the incense stick cordycep mainly contains following several:
1. volumetric method: with periodic acid sweet mellow wine is oxidized to formaldehyde, the acid iodide of generation and excessive periodic acid are to the potassium iodide effect, and the iodine that dissociates out uses the difference of sodium thiosulfate titration and blank value to obtain content;
2. colourimetry: sodium metaperiodate and sweet mellow wine reaction produce yellow 3,5-diacetyl-1,4 dehydrogenation lutidines, and this compound is the absorption maximum when 412nm, so the survey of available colourimetry;
3. thin layer chromatography scanning: Wang Baoqi etc. filter sample as need testing solution with anhydrous alcohol solution after with 95% alcohol reflux, [No. 1 is dioxane-isoamyl acetate-isopropyl alcohol-water (5:15:5:2) with No. 1 developping agent-No. 2 developping agent-glacial acetic acid (60:40:1) ascending development, No. 2 is isopropyl alcohol-ethyl acetate-water (54:7:8)], launch 0.12mol/L hydrochloric acid solution and the 0.5% biphenylamine ethanolic solution of back with the potassium metaperiodate of 0.015mol/L, at λ s=295nm, measure under the λ R=370nm condition;
Mensuration to sweet mellow wine, common method has volumetric method and colourimetry, but the drawback of volumetric method is to only limit to pure product measure, if the aggregate of preparation or multiple compound, then reducing substances also can be by periodate oxidation in the component, and the result makes the content that records more higher than actual value; And that the shortcoming of colourimetry is a specificity is poor, easily produces error; Because the content of sweet mellow wine is all different in the incense stick cordycep of various places, so measure with thin layer chromatography scanning, error is bigger.
Because the content of sweet mellow wine is all different in the incense stick cordycep of various places, for the distinctive incense stick cordycep in Shanxi, above several method all can not efficiently and accurately the content that determines its sweet mellow wine, in order to address this problem the sweet mellow wine that the present invention contains with the peculiar incense stick cordycep in gas chromatography determination Shanxi.
Summary of the invention
The detection method that the purpose of this invention is to provide mannitol content in a kind of incense stick cordycep, the present invention is the content that utilizes sweet mellow wine in the gas chromatography determination incense stick cordycep, to reach the content that determines sweet mellow wine in the distinctive incense stick cordycep in Shanxi of efficiently and accurately.
The detection method of mannitol content in the incense stick cordycep of the present invention may further comprise the steps:
(1) preparation of reference substance solution: precision takes by weighing 100 ℃ of sweet mellow wine reference substance 30mg that are dried to constant weight in the 10ml volumetric flask, adding volume ratio is pyridine-aceticanhydride 5ml of 1:1, acetylization reaction 40min in 100 ℃ of water-baths, put cold back and be settled to scale with pyridine, shake up, promptly get reference substance solution;
(2) preparation of need testing solution: with the incense stick cordycep crushing screening, precision takes by weighing 1g and adds absolute ethyl alcohol extract 5.5-6.5 hour to closely colourless in apparatus,Soxhlet's, extract is recycled to dried, residue adds the pyridine that volume ratio is 1:1-aceticanhydride 40ml, acetylation 40min in 100 ℃ of water-baths, put to move in the 50ml volumetric flask after cold and be settled to scale, shake up, promptly get need testing solution with pyridine;
(3) adopt gas chromatograph to measure, chromatographic condition is: the initial temperature of column temperature is 220 ℃, keeps 1min. to rise to 220 ℃-230 ℃ with the speed of 10 ℃/min, keeps 26min, and vaporizer and flame ionization ditector temperature are 300 ℃, H
2Flow velocity 34-40ml/min, air velocity 350-400ml/min, N
2Flow velocity 30-35/ min, split ratio 50:1.
Further, described H
2Flow velocity 34.3ml/min, N
2Flow velocity 33ml/min, air velocity 351.9ml/min.
The accurate respectively reference substance solution 0.5,1,1.5,2 of drawing, 2.5ml is in the 5ml volumetric flask, be settled to scale with pyridine, shake up, last gas chromatograph for determination, sample size 1 μ l, with concentration is horizontal ordinate, and peak area is an ordinate drawing standard curve, the results are shown in Table 1 and Fig. 1
Table 1
| The amount of sweet mellow wine (μ g) | A |
| 0.3377 | 90289 |
| 0.6754 | 169655 |
| 1.0131 | 263084 |
| 1.3508 | 352923 |
| 1.6885 | 450137 |
Show that sweet mellow wine has good linear relationship between 0.3377～1.6885 μ g.
Select methyl alcohol respectively, ethanol, Shui and absolute ethyl alcohol are as solvent, and it is as shown in table 2 to extract the back mannitol content,
The different extraction solvent of table 2 are to the influence of mannitol content
| Extraction solvent | Mannitol content (mg/g) |
| 50% methyl alcohol | 38.2 |
| 50% ethanol | 40.8 |
| Water | 16.2 |
| Absolute ethyl alcohol | 56.7 |
The result shows that the absolute ethyl alcohol extraction efficiency is the highest, finally is defined as with absolute ethyl alcohol as extraction solvent.
Carry out accuracy test according to the inventive method, adopt the application of sample absorption method, the result is as shown in table 3,
Table 3
From last Biao Kede, the inventive method accuracy is good.
Advantage of the present invention is easy, quick, accurate, favorable reproducibility, more science, measure mannitol content in the distinctive incense stick cordycep in Shanxi exactly, improve the quality of product effectively.
Description of drawings
Fig. 1 is the sweet mellow wine canonical plotting.
Embodiment
The detection method of mannitol content in the incense stick cordycep may further comprise the steps:
(1) preparation of reference substance solution: precision takes by weighing 100 ℃ of sweet mellow wine reference substance 30mg that are dried to constant weight in the 10ml volumetric flask, adding volume ratio is pyridine-aceticanhydride 5ml of 1:1, acetylization reaction 40min in 100 ℃ of water-baths, put cold back and be settled to scale with pyridine, shake up, promptly get reference substance solution;
(2) preparation of need testing solution: with the incense stick cordycep crushing screening, precision takes by weighing 1g and adds absolute ethyl alcohol extract 5.5 hours to closely colourless in apparatus,Soxhlet's, extract is recycled to dried, residue adds the pyridine that volume ratio is 1:1-aceticanhydride 40ml, acetylation 40min in 100 ℃ of water-baths, put to move in the 50ml volumetric flask after cold and be settled to scale, shake up, promptly get need testing solution with pyridine;
(3) adopt gas chromatograph to measure, chromatographic condition is: the initial temperature of column temperature is 220 ℃, keeps 1min. to rise to 220 ℃ ℃ with the speed of 10 ℃/min, keeps 26min, and vaporizer and flame ionization ditector temperature are 300 ℃, H
2Flow velocity 40ml/min, air velocity 350ml/min, N
2Flow velocity 30/ min, split ratio 50:1.
The detection method of mannitol content in the incense stick cordycep may further comprise the steps:
(1) preparation of reference substance solution: precision takes by weighing 100 ℃ of sweet mellow wine reference substance 30mg that are dried to constant weight in the 10ml volumetric flask, adding volume ratio is pyridine-aceticanhydride 5ml of 1:1, acetylization reaction 40min in 100 ℃ of water-baths, put cold back and be settled to scale with pyridine, shake up, promptly get reference substance solution;
(2) preparation of need testing solution: with the incense stick cordycep crushing screening, precision takes by weighing 1g and adds absolute ethyl alcohol extract 6.5 hours to closely colourless in apparatus,Soxhlet's, extract is recycled to dried, residue adds the pyridine that volume ratio is 1:1-aceticanhydride 40ml, acetylation 40min in 100 ℃ of water-baths, put to move in the 50ml volumetric flask after cold and be settled to scale, shake up, promptly get need testing solution with pyridine;
(3) adopt gas chromatograph to measure, chromatographic condition is: the initial temperature of column temperature is 220 ℃, keeps 1min. to rise to 230 ℃ ℃ with the speed of 10 ℃/min, keeps 26min, and vaporizer and flame ionization ditector temperature are 300 ℃, H
2Flow velocity 34ml/min, air velocity 400ml/min, N
2Flow velocity 35/ min, split ratio 50:1.
Embodiment 3
The detection method of mannitol content in the incense stick cordycep may further comprise the steps:
(1) preparation of reference substance solution: precision takes by weighing 100 ℃ of sweet mellow wine reference substance 30mg that are dried to constant weight in the 10ml volumetric flask, adding volume ratio is pyridine-aceticanhydride 5ml of 1:1, acetylization reaction 40min in 100 ℃ of water-baths, put cold back and be settled to scale with pyridine, shake up, promptly get reference substance solution;
(2) preparation of need testing solution: with the incense stick cordycep crushing screening, precision takes by weighing 1g and adds absolute ethyl alcohol extract 6 hours to closely colourless in apparatus,Soxhlet's, extract is recycled to dried, residue adds the pyridine that volume ratio is 1:1-aceticanhydride 40ml, acetylation 40min in 100 ℃ of water-baths, put to move in the 50ml volumetric flask after cold and be settled to scale, shake up, promptly get need testing solution with pyridine;
(3) adopt gas chromatograph to measure, chromatographic condition is: the initial temperature of column temperature is 220 ℃, keeps 1min. to rise to 225 ℃ ℃ with the speed of 10 ℃/min, keeps 26min, and vaporizer and flame ionization ditector temperature are 300 ℃, H
2Flow velocity 34.3ml/min, N
2Flow velocity 33ml/min, air velocity 351.9ml/min, split ratio 50:1.
Claims (2)
1. the detection method of mannitol content in the incense stick cordycep is characterized in that may further comprise the steps:
(1) preparation of reference substance solution: precision takes by weighing 100 ℃ of sweet mellow wine reference substance 30mg that are dried to constant weight in the 10ml volumetric flask, adding volume ratio is pyridine-aceticanhydride 5ml of 1:1, acetylization reaction 40min in 100 ℃ of water-baths, put cold back and be settled to scale with pyridine, shake up, promptly get reference substance solution;
(2) preparation of need testing solution: with the incense stick cordycep crushing screening, precision takes by weighing 1g and adds absolute ethyl alcohol extract 5.5-6.5 hour to closely colourless in apparatus,Soxhlet's, extract is recycled to dried, residue adds the pyridine that volume ratio is 1:1-aceticanhydride 40ml, acetylation 40min in 100 ℃ of water-baths, put to move in the 50ml volumetric flask after cold and be settled to scale, shake up, promptly get need testing solution with pyridine;
(3) adopt gas chromatograph to measure, chromatographic condition is: the initial temperature of column temperature is 220 ℃, keeps 1min. to rise to 220 ℃-230 ℃ with the speed of 10 ℃/min, keeps 26min, and vaporizer and flame ionization ditector temperature are 300 ℃, H
2Flow velocity 34-40ml/min, air velocity 350-400ml/min, N
2Flow velocity 30-35/ min, split ratio 50:1.
2. the detection method of mannitol content is characterized in that described H in the incense stick cordycep according to claim 1
2Flow velocity 34.3ml/min, N
2Flow velocity 33ml/min, air velocity 351.9ml/min.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517083A (en) * | 2018-11-01 | 2019-03-26 | 佛山科学技术学院 | The preparation method and applications of Cordyceps hawkesii Gary intracellular polyse |
| CN110133129A (en) * | 2019-05-15 | 2019-08-16 | 江西国药有限责任公司 | The detection method of mannitol in a kind of fermentation cordyceps (Cs-4) |
| CN110672782A (en) * | 2019-10-18 | 2020-01-10 | 武汉工程大学 | Method for determining and analyzing glycols in levophosphorus dextroamine salt |
-
2011
- 2011-03-23 CN CN 201110070678 patent/CN102183593B/en active Active
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| T.IVERSEN等: "Lignin-carbohydrate bonds in a lignin-carbohydrate complex isolated from spruce", 《WOOD SCIENCE AND TECHNOLOGY》 * |
| 刘敬兰等: "衍生化毛细管气相色谱法分离测定山梨醇和甘露醇", 《分析化学》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517083A (en) * | 2018-11-01 | 2019-03-26 | 佛山科学技术学院 | The preparation method and applications of Cordyceps hawkesii Gary intracellular polyse |
| CN110133129A (en) * | 2019-05-15 | 2019-08-16 | 江西国药有限责任公司 | The detection method of mannitol in a kind of fermentation cordyceps (Cs-4) |
| CN110672782A (en) * | 2019-10-18 | 2020-01-10 | 武汉工程大学 | Method for determining and analyzing glycols in levophosphorus dextroamine salt |
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Address after: 030031 No. 85, Longcheng street, Xiaodian District, Taiyuan City, Shanxi Province Patentee after: SHANXI INSTITUTE OF FOOD AND DRUG INSPECTION (SHANXI PHARMACEUTICAL PACKAGING MATERIAL MONITORING CENTER) Country or region after: China Address before: 030001 No. 12 Taoyuan South Road, Shanxi, Taiyuan Patentee before: Shanxi Institute for Food and Drug Control Country or region before: China |
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