CN101732364A - Method for identifying lucid ganoderma alkaloids substance finger-print - Google Patents
Method for identifying lucid ganoderma alkaloids substance finger-print Download PDFInfo
- Publication number
- CN101732364A CN101732364A CN200910154774A CN200910154774A CN101732364A CN 101732364 A CN101732364 A CN 101732364A CN 200910154774 A CN200910154774 A CN 200910154774A CN 200910154774 A CN200910154774 A CN 200910154774A CN 101732364 A CN101732364 A CN 101732364A
- Authority
- CN
- China
- Prior art keywords
- sample
- testing sample
- 1vol
- acetic acid
- testing
- 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.)
- Granted
Links
- 241000222336 Ganoderma Species 0.000 title claims abstract description 93
- 229930013930 alkaloid Natural products 0.000 title claims abstract description 79
- 239000000126 substance Substances 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title claims abstract description 32
- 238000012360 testing method Methods 0.000 claims abstract description 82
- 239000012634 fragment Substances 0.000 claims abstract description 24
- 239000003814 drug Substances 0.000 claims abstract description 19
- 238000004458 analytical method Methods 0.000 claims abstract description 14
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 120
- 239000000523 sample Substances 0.000 claims description 120
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 90
- 239000000284 extract Substances 0.000 claims description 43
- 239000007791 liquid phase Substances 0.000 claims description 33
- 239000012488 sample solution Substances 0.000 claims description 31
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 26
- 240000008397 Ganoderma lucidum Species 0.000 claims description 25
- 235000001637 Ganoderma lucidum Nutrition 0.000 claims description 25
- 238000001035 drying Methods 0.000 claims description 22
- 238000010992 reflux Methods 0.000 claims description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 18
- 150000003797 alkaloid derivatives Chemical class 0.000 claims description 18
- 229940079593 drug Drugs 0.000 claims description 17
- 239000012071 phase Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 11
- 238000010828 elution Methods 0.000 claims description 10
- 150000002825 nitriles Chemical class 0.000 claims description 10
- 238000004587 chromatography analysis Methods 0.000 claims description 9
- 230000006837 decompression Effects 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 9
- 238000004821 distillation Methods 0.000 claims description 9
- 239000000945 filler Substances 0.000 claims description 9
- 238000005374 membrane filtration Methods 0.000 claims description 9
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 9
- 239000000377 silicon dioxide Substances 0.000 claims description 9
- 238000000605 extraction Methods 0.000 claims description 5
- 238000010606 normalization Methods 0.000 claims description 5
- 238000005303 weighing Methods 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 238000005520 cutting process Methods 0.000 claims description 2
- 235000013399 edible fruits Nutrition 0.000 abstract 2
- 238000010521 absorption reaction Methods 0.000 abstract 1
- 241000207961 Sesamum Species 0.000 description 15
- 235000003434 Sesamum indicum Nutrition 0.000 description 15
- 238000011156 evaluation Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 230000014759 maintenance of location Effects 0.000 description 6
- 235000007215 black sesame Nutrition 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000002994 raw material Substances 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000008821 health effect Effects 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000000874 microwave-assisted extraction Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 208000017897 Carcinoma of esophagus Diseases 0.000 description 1
- 206010010144 Completed suicide Diseases 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 208000005016 Intestinal Neoplasms Diseases 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 1
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000000954 anitussive effect Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000001088 anti-asthma Effects 0.000 description 1
- 239000000924 antiasthmatic agent Substances 0.000 description 1
- 229940124584 antitussives Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 125000000567 diterpene group Chemical group 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 201000002313 intestinal cancer Diseases 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 201000000498 stomach carcinoma Diseases 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a method for identifying lucid ganoderma alkaloids substance finger-print, comprising the following steps: adopting 5-10 batches of dried lucid ganoderma fruit body fragments from known standard variety samples produced in different places; taking the same mass of crude medicine in each batch for producing test article solution; and carrying out high performance liquid chromatography analysis on the test article solution; taking common absorption peaks of the high performance liquid chromatography as characteristic chromatographic peaks; choosing chromatographic peaks with uniformization peak area exceeding 1% as fingerprint peaks; obtaining standard lucid ganoderma alkaloids substance finger-print; taking the sample to be tested: dried lucid ganoderma fruit body fragments, obtaining high performance liquid chromatography of the sample to be tested: the lucid ganoderma alkaloids substance by the same method. The high performance liquid chromatography of the sample to be tested: the lucid ganoderma alkaloids substance has characteristic peaks found in the standard finger-print of the lucid ganoderma alkaloids substance, the sample to be tested is a standard variety. With the method of the invention adopted, different lucid ganoderma varieties can be identified.
Description
Technical field
The invention belongs to edible mushroom and Chinese medicine raw material alkaloids substance quality analyzing technique and identify the field, be specifically related to construction method and the analysis and identification method of a kind of efficient liquid-phase chromatograph finger print atlas of the biological active component alkaloids substance in the glossy ganoderma.
Background technology
Glossy ganoderma (Ganoderma Lucidum) is described as the miraculous cure that the mankind promote longevity since ancient times, nourishes the celestial grass of beneficial gas. In China's this agrostology of First monograph Shennong's Herbal, just think that it has the beneficial motive, pacifies smart soul, the effect of tonifying liver benefit gas, hard muscles and bones. Li Shizhen (1518-1593 A.D.) in Compendium of Material Medica also the food therapy health effect to glossy ganoderma done high evaluation: " red sesame has another name called red sesame, and smell is bitter flat, and is nontoxic. Cure mainly in the heart knot, the beneficial motive, bowl spares increases wisdom, does not forget. Food of a specified duration is made light of one's life by commiting suicide not old, and the angle prolongs life ". In recent decades, scientist is by various researchs, and prove from many aspects: glossy ganoderma has very the disease-relieving and physique enhancing of " spirit ", the health-care effect of strengthening the body resistance to consolidate the constitution. Especially biological immune and traditional treatment tumour had peculiar effect.
A large amount of chemistry and pharmacology analysis show in recent years: glossy ganoderma specifically have antitumor, alleviate pain that cancer or other diseases cause, strengthen immunity, delay senility, the multiple effects such as protecting liver and detoxication, treatment hypertension, hypoglycemic, antitussive and antiasthmatic, radioresistance and antiallergy. Especially at anti-tumor aspect, many famous medical research mechanisms reach a conclusion after deliberation both at home and abroad: glossy ganoderma has very significant curative effect to kinds of tumors diseases such as cancer of the stomach, lung cancer, malignant lymphoma, liver cancer, breast cancer, nasopharyngeal carcinoma, intestinal cancer, cancer of the esophagus, leukaemia. Why glossy ganoderma has so significant medical health effect, in the active material wherein, lucid ganoderma alkaloids (such as Diterpenes, triterpenes, fast quinoline alkali, unknown alkali etc.) is a kind of important basic effective component in the glossy ganoderma, and its effect mainly comprises: protect the liver; Antitumor; Hypoglycemic; Anti-HIV-1 and HIV-1 proteinase activity; Suppress histamine release; Angiotensin-converting enzyme inhibition; Suppress the effects such as cholesterol biosynthesis. Therefore, lucid ganoderma alkaloids has important medicines and health protection and is worth, and glossy ganoderma has been developed to the plurality kinds of health care product by different processing modes at present, and lucid ganoderma alkaloids substance is a kind of product wherein.
Present domestic glossy ganoderma kind has a lot, such as red sesame, black sesame, purple sesame, Han Zhi, Bai Zhi etc., wherein in the majority with red sesame kind, because cultivation of glossy ganoderma is different with difference, planting type, the kind in weather conditions, zone, the biological active component alkaloids substance has certain difference in the glossy ganoderma, how to differentiate biological active component alkaloids substance contained in the glossy ganoderma, the quality of glossy ganoderma is differentiated and scientific evaluation has the meaning of value lever. At present, to different glossy ganoderma kinds, with the kind zones of different, with the discriminating of the biological active component alkaloids substance of the different cultural methods of kind and the evaluation method that a science is not also set up in quality evaluation, there is not the method report of related fields yet.
Summary of the invention
The objective of the invention is in order to set up the analysis and identification method of glossy ganoderma raw material alkaloids substance, preliminary qualitative, quantitative attributional analysis to glossy ganoderma alkaloid processing quality control, the discriminating of the raw material true and false and alkaloids substance provides a kind of effective method, thereby the material quality of glossy ganoderma is carried out the evaluation of science.
The technical solution used in the present invention is:
A kind of method for identifying lucid ganoderma alkaloids substance finger-print, described method may further comprise the steps:
(a) producing of standard finger-print: 5～10 batches, the every batch crude drug amount of getting equal in quality of ganoderma lucidum fruitbody fragment of getting the different known standard variety sample drying in the place of production is carried out respectively following test: the alcohol reflux that adds 25～35 times of quality of ganoderma lucidum fruitbody fragment of known sample drying extracts, obtain the known sample extract after removing by filter filter residue, the known sample extract is through the decompression distillation desolventizing, get known sample alkaloid medicinal extract liquid, add methanol constant volume and get known sample solution, controlling in the described known sample solution crude drug amount content is/100 milliliters of 2～7 grams, known sample solution is through the organic membrane filtration of 0.45 μ m, get the known sample need testing solution, get the known sample need testing solution and carry out efficient liquid phase chromatographic analysis, obtain known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram; The condition of described high performance liquid chromatography is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, and the detection wavelength is 254nm; Get that the total absworption peak of high-efficient liquid phase chromatogram is Characteristic chromatographic peak in each batch, select among the figure normalization peak area greater than 1% Characteristic chromatographic peak as fingerprint peaks, obtain the standard finger-print of lucid ganoderma alkaloids substance;
(b) the testing sample high-efficient liquid phase chromatogram is produced: the ganoderma lucidum fruitbody fragment of getting the testing sample drying, the alcohol reflux that adds 25～35 times of quality of ganoderma lucidum fruitbody fragment of testing sample drying extracts, obtain the testing sample extract after removing by filter filter residue, the testing sample extract is through the decompression distillation desolventizing, get testing sample alkaloid medicinal extract liquid, add methanol constant volume and get testing sample solution, it is identical to control in the described testing sample solution in the crude drug amount content and step (a) the crude drug amount content of known sample solution, testing sample solution is through the organic membrane filtration of 0.45 μ m, get the testing sample need testing solution, get the testing sample need testing solution and carry out efficient liquid phase chromatographic analysis, the condition of described high performance liquid chromatography is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, the detection wavelength is 254nm, obtains testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram;
Characteristic chromatographic peak contrast in the standard finger-print of the lucid ganoderma alkaloids substance that the testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram that (c) step (b) is obtained and step (a) obtain, testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram has feature chromatogram cutting edges of a knife or a sword all in the standard finger-print of lucid ganoderma alkaloids substance, and then testing sample is standard variety.
Comparatively concrete, in the producing of described step (a) standard finger-print, the operation that every batch of ganoderma lucidum fruitbody fragment of getting the known standard variety sample drying of equal in quality crude drug amount makes known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram may further comprise the steps:
(1) takes by weighing the ganoderma lucidum fruitbody fragment of known sample drying, add 95wt% ethanol, the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and known sample drying is 25～35: 1, preferred 33: 1, under little reflux state that boils, extracted 1～2 hour, remove by filter filter residue after, get the known sample extract;
(2) the known sample extract is through the decompression distillation desolventizing, get known sample alkaloid medicinal extract liquid, add methanol constant volume and get known sample solution, controlling in the described known sample solution crude drug amount content is/100 milliliters of 2～7 grams, known sample solution gets the known sample need testing solution through the organic membrane filtration of 0.45 μ m;
(3) the known sample need testing solution carries out efficient liquid phase chromatographic analysis, get known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram, described chromatographic condition is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, flow rate of mobile phase is 1mL/min, the detection wavelength is 254nm, sample size 10 μ L, be 40min analysis time, the gradient elution program is: 0～8min, the 1vol% aqueous acetic acid of 10vol% and the acetonitrile of 90vol%; 8～10min, the 1vol% aqueous acetic acid of 15vol% and the acetonitrile of 85vol%; 10～15min, the 1vol% aqueous acetic acid of 30vol% and the acetonitrile of 70vol%; 15～20min, the 1vol% aqueous acetic acid of 40vol% and the acetonitrile of 60vol%; 20～30min, the 1vol% aqueous acetic acid of 45vol% and the acetonitrile of 55vol%; 30～40min, the 1vol% aqueous acetic acid of 50vol% and the acetonitrile of 50vol%.
Comparatively concrete, the operation that the testing sample high-efficient liquid phase chromatogram is produced in the described step (b) may further comprise the steps:
(1) takes by weighing the ganoderma lucidum fruitbody fragment of testing sample drying, add 95wt% ethanol, the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and testing sample drying is 25～35: 1, preferred 33: 1, under little reflux state that boils, extracted 1～2 hour, remove by filter filter residue after, get the testing sample extract;
(2) the testing sample extract is through the decompression distillation desolventizing, get testing sample alkaloid medicinal extract liquid, add methanol constant volume and get testing sample solution, it is identical to control in the described testing sample solution in the crude drug amount content and step (a) the crude drug amount content of known sample solution, testing sample solution gets the testing sample need testing solution through the organic membrane filtration of 0.45 μ m;
(3) the testing sample need testing solution carries out efficient liquid phase chromatographic analysis, get high-efficient liquid phase chromatogram, described chromatographic condition is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, flow rate of mobile phase is 1mL/min, and the detection wavelength is 254nm, sample size 10 μ L, be 40min analysis time, and the gradient elution program is: 0～8min, the 1vol% aqueous acetic acid of 10vol% and the acetonitrile of 90vol%; 8～10min, the 1vol% aqueous acetic acid of 15vol% and the acetonitrile of 85vol%; 10～15min, the 1vol% aqueous acetic acid of 30vol% and the acetonitrile of 70vol%; 15～20min, the 1vol% aqueous acetic acid of 40vol% and the acetonitrile of 60vol%; 20～30min, the 1vol% aqueous acetic acid of 45vol% and the acetonitrile of 55vol%; 30～40min, the 1vol% aqueous acetic acid of 50vol% and the acetonitrile of 50vol%.
The ganoderma lucidum fruitbody fragment of known sample drying of the present invention be with known sample ganoderma lucidum fruitbody section, dry, be crushed to 40～80 orders and obtain.
The ganoderma lucidum fruitbody fragment of testing sample drying of the present invention be with testing sample ganoderma lucidum fruitbody section, dry, be crushed to 40～80 orders and obtain.
Of the present invention under little reflux state that boils, extracted 1～2 hour, remove by filter filter residue after, get the known sample extract, be preferably under little reflux state that boils, extracted 1 hour, remove by filter filter residue after, get the known sample extract; More preferably: adopting Microwave-assisted Extraction to follow the example of, is under the microwave condition of 800W at power, and microwave auxiliary reflux extracts 17min, remove by filter filter residue after, get the known sample extract.
Of the present invention under little reflux state that boils, extracted 1～2 hour, remove by filter filter residue after, get the testing sample extract, be preferably under little reflux state that boils, extracted 1 hour, remove by filter filter residue after, get the testing sample extract; More preferably: adopting Microwave-assisted Extraction to follow the example of, is under the microwave condition of 800W at power, and microwave auxiliary reflux extracts 17min, remove by filter filter residue after, get the testing sample extract.
The present invention can be to different glossy ganoderma kinds, with the different places of production of kind, carry out discriminatory analysis with the biological active component alkaloids substance of kind different planting, can be with reference to collection of illustrative plates by the alkaloid chromatogram of setting a certain lucidum variety, coupling and calculated by peak area by similarity evaluation, automatically calculate the similarity that has peak composition and sample in the red ganoderma sample of different producing regions, calculated value with reference to size and the similarity of peak area, carry out data analysis and can more scientifically differentiate the contained alkaloidal quality situation of different producing regions red ganoderma, thereby differentiate that tentatively which kind of different classes of kind has more similitude and otherness, the quality of contained alkaloids is better.
The present invention preferably carries out pre-treatment with the microwave auxiliary reflux extraction method, optimized the alkaloid extracting method, adopt the C18 post, carry out take own nitrile, 1vol% aqueous acetic acid as mobile phase the binary gradient elution fingerprint map analyzing of alkaloids substance in the glossy ganoderma is differentiated have quick, easy, stable, directly perceived, favorable reproducibility, with strong points, alkaloid composes that the peak number order is many, the advantage of good separating effect and easy operating. Use the inventive method to different glossy ganoderma kinds, with the different places of production of kind, ideal with the identification result of the biological active component alkaloids substance of kind different planting, the quality of the biological active component alkaloids substance of glossy ganoderma is differentiated the scientific basis that has proposed first preliminary qualitative, quantitative.
Figure of description
The finger-print of alkaloids substance in 1 No. a kind red ganodermas of accompanying drawing sample
The alkaloids substance chromatographic peak contrast of 29 kinds of different producing regions of accompanying drawing red ganoderma sample refers to collection of illustrative plates, the total absworption peak of getting these 9 kinds of high-efficient liquid phase chromatograms is Characteristic chromatographic peak, select among the figure normalization peak area greater than 1% Characteristic chromatographic peak as fingerprint peaks, obtain the standard finger-print of red ganoderma alkaloids substance.
35 kinds of red sesames of different genera of accompanying drawing, Han Zhi, purple sesame, black sesame, the contrast of Wild ganoderma alkaloids substance chromatographic peak refer to that collection of illustrative plates, the red sesame of red expression, Korea Spro represent Han Zhi, the purple sesame of purple expression, the black sesame of black expression, wild expression Wild ganoderma.
The specific embodiment:
The below further specifies the present invention with specific embodiment, but protection scope of the present invention is not limited to this.
Embodiment 1 sets up the finger-print of alkaloids substance in the glossy ganoderma
1. pulverising step: with No. a kind red ganoderma sample entity section, dry, be crushed to about 40 orders, get the glossy ganoderma dry product.
2. extraction step: take by weighing 5g glossy ganoderma dry product, add 95wt% ethanol at 1: 33 with solid-liquid ratio, under little reflux state that boils, adopt 800W power, microwave refluxing extraction 17min, remove by filter filter residue after, get extract.
3. extract gets alkaloid medicinal extract liquid through the decompression distillation desolventizing, adds methanol constant volume to 100mL, gets sample solution, and red ganoderma dry product content is/100 milliliters of 5 grams in the sample solution: sample solution gets need testing solution through the organic membrane filtration of 0.45 μ m;
4. need testing solution carries out efficient liquid phase chromatographic analysis, gets high-efficient liquid phase chromatogram. Chromatographic condition is: the Waters1525 high performance liquid chromatograph, and binary pump, UV-detector, Bress software, octadecylsilane chemically bonded silica are the chromatographic column C of filler18(model: Symmetry250 * 4.6mm I.D.5 μ m, U.S. Waters company produces), carry out gradient elution take own nitrile, water (containing 1% acetic acid v/v) as mobile phase, flow rate of mobile phase is 1mL/min, the detection wavelength is 254nm, sample size 10 μ L, and be 40 minutes analysis time, the gradient elution program is: 0～8min, the 1vol% aqueous acetic acid of 10vol% and the acetonitrile of 90vol%; 8～10min, the 1vol% aqueous acetic acid of 15vol% and the acetonitrile of 85vol%; 10～15min, the 1vol% aqueous acetic acid of 30vol% and the acetonitrile of 70vol%; 15～20min, the 1vol% aqueous acetic acid of 40vol% and the acetonitrile of 60vol%; 20～30min, the 1vol% aqueous acetic acid of 45vol% and the acetonitrile of 55v ol%; 30～40min, the 1vol% aqueous acetic acid of 50vol% and the acetonitrile of 50vol%.
5. fingerprint map construction: in high-efficient liquid phase chromatogram, select the normalization peak area greater than 1% chromatographic peak as fingerprint peaks, finally selected 38 fingerprint peakses (accounting for more than 90% of total peak area) to analyze collection of illustrative plates as the unimodal alkaloids of glossy ganoderma, see Fig. 1, be fingerprint peaks among the figure shown in the bullet. But have so the alkaloidal kind of glossy ganoderma in the qualitative discrimination red ganoderma sample, simultaneously according to the normalized area computing method, roughly differentiate accordingly and go out peak retention time section, which kind of alkaloid is higher.
The preliminary qualitative, quantitative of embodiment 2 different producing regions red ganoderma alkaloids is discriminatory analysis relatively
Gather 9 kinds of different producing regions red ganoderma samples and carry out the alkaloids analysis, experiment condition and method of operating are with embodiment 1, obtain 9 kinds of red ganoderma sample alkaloids and analyze collection of illustrative plates, then adopt similarity evaluation, choose a kind of producing region red ganoderma sample as reference spectrum peak (R), and carry out the method for chromatographic peak Supplements, the chromatographic peak of 8 samples mated and automatically generate contrast refer to collection of illustrative plates, see accompanying drawing 2, coupling and calculated by peak area by similarity evaluation, automatically calculate in the red ganoderma sample of different producing regions that total peak forms and the similarity of sample, see Table respectively 1, table 2.
Logarithmic data is analyzed in table 1 chromatographic peak peak area and total peak match
| Retention time | S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | R | Total peak number |
| 1.492 | 0 | 469.427 | 0 | 0 | 203.16 | 0 | 257.59 | 217.35 | 143.44 | 4 |
| 2.158 | 259.08 | 229.414 | 206.24 | 228.47 | 0 | 0 | 0 | 0 | 115.40 | 4 |
| 2.333 | 201.45 | 332.056 | 0 | 0 | 0 | 0 | 0 | 224.21 | 94.716 | 3 |
| 2.664 | 1576.5 | 2298.32 | 0 | 1882.2 | 1056.3 | 926.53 | 1530.7 | 1086.3 | 1294.6 | 7 |
| 3.537 | 1052.3 | 865.414 | 260.47 | 631.50 | 268.89 | 218.51 | 438.23 | 311.83 | 505.90 | 8 |
| 3.750 | 292.33 | 342.329 | 380.33 | 365.44 | 271.29 | 319.38 | 204.65 | 387.19 | 320.37 | 8 |
| 12.639 | 248.16 | 200.184 | 0 | 0 | 222.18 | 0 | 0 | 0 | 83.817 | 3 |
| 12.979 | 443.71 | 229.103 | 207.50 | 281.01 | 277.57 | 323.54 | 357.94 | 340.33 | 307.59 | 8 |
| 14.292 | 229.89 | 0 | 0 | 0 | 233.11 | 0 | 230.23 | 204.45 | 112.21 | 4 |
| 16.29 | 455.87 | 0 | 214.41 | 0 | 0 | 238.14 | 224.23 | 226.07 | 169.84 | 5 |
| 16.717 | 454.61 | 0 | 0 | 0 | 318.90 | 0 | 214.28 | 0 | 123.47 | 3 |
| 17.461 | 357.81 | 202.328 | 325.69 | 0 | 0 | 317.27 | 221.19 | 367.69 | 224.00 | 6 |
| 17.828 | 561.75 | 245.117 | 392.43 | 243.89 | 0 | 388.3 | 262.60 | 430.23 | 315.54 | 7 |
| 18.158 | 542.31 | 241.559 | 541.21 | 313.94 | 205.68 | 639.38 | 489.51 | 642.99 | 452.07 | 8 |
| 19.2 | 328.47 | 251.271 | 272.29 | 0 | 398.67 | 247.59 | 0 | 296.24 | 224.31 | 6 |
| 19.386 | 569.96 | 212.4 | 276.72 | 0 | 0 | 204.98 | 257.51 | 294.50 | 227.01 | 6 |
| 19.902 | 1299.1 | 790.102 | 1360.3 | 558.44 | 510.30 | 1368.7 | 652.80 | 1456.8 | 999.60 | 8 |
| 20.156 | 784.99 | 391.413 | 713.78 | 323.32 | 366.75 | 692.95 | 526.62 | 794.47 | 574.28 | 8 |
| Retention time | S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | R | Total peak number |
| 20.513 | 0 | 273.604 | 220.21 | 0 | 349.19 | 237.25 | 202.39 | 240.45 | 162.86 | 5 |
| 20.767 | 713.02 | 345.269 | 694.81 | 286.20 | 0 | 440.49 | 274.63 | 493.03 | 405.93 | 7 |
| 20.945 | 330.86 | 220.211 | 276.75 | 0 | 211.94 | 261.02 | 282.13 | 312.76 | 236.96 | 7 |
| 21.193 | 460.36 | 438.747 | 812.60 | 213.17 | 0 | 757.63 | 399.18 | 399.18 | 502.82 | 7 |
| 21.725 | 0 | 0 | 0 | 281.22 | 0 | 410.55 | 362.09 | 2034.0 | 385.98 | 4 |
| 21.805 | 548.94 | 342.893 | 0 | 0 | 0 | 1172.3 | 0 | 0 | 258.02 | 3 |
| 21.864 | 978.85 | 786.401 | 1611.2 | 328.31 | 520.39 | 0 | 628.33 | 0 | 606.68 | 6 |
| 22.571 | 202.39 | 0 | 0 | 0 | 249.58 | 0 | 209.33 | 204.57 | 108.23 | 4 |
| 22.933 | 258.10 | 0 | 204.16 | 0 | 0 | 214.48 | 0 | 0 | 84.594 | 3 |
| 23.916 | 253.83 | 307.435 | 228.67 | 0 | 590.15 | 0 | 0 | 275.26 | 206.92 | 5 |
| 24.238 | 469.04 | 407.614 | 671.56 | 0 | 203.71 | 569.73 | 244.59 | 760.22 | 415.81 | 7 |
| 24.847 | 523.59 | 457.65 | 563.65 | 0 | 283.83 | 479.07 | 219.82 | 640.13 | 395.97 | 7 |
| 25.33 | 320.14 | 0 | 323.08 | 0 | 248.33 | 221.79 | 241.58 | 303.52 | 207.30 | 6 |
| 25.669 | 217.50 | 293.771 | 207.22 | 0 | 621.66 | 356.39 | 310.96 | 282.77 | 286.28 | 7 |
| 27.166 | 388.86 | 303.588 | 295.68 | 0 | 280.62 | 271.10 | 0 | 370.27 | 238.76 | 6 |
| 19293. | 12530。 | 12615。 | 6172. | 8537. | 11277. | 9462.1 | 14617. | 113813 |
Annotate: Sx is the glossy ganoderma sample type
Table 2 chromatographic peak similarity value calculation
| S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | R | |
| S1 | 1 | 0.812 | 0.765 | 0.745 | 0.698 | 0.841 | 0.831 | 0.721 | 0.934 |
| S2 | 0.812 | 1 | 0.505 | 0.897 | 0.774 | 0.884 | 0.883 | 0.677 | 0.916 |
| S3 | 0.765 | 0.505 | 1 | 0.364 | 0.495 | 0.579 | 0.654 | 0.565 | 0.731 |
| S4 | 0.745 | 0.897 | 0.364 | 1 | 0.665 | 0.874 | 0.859 | 0.598 | 0.847 |
| S5 | 0.698 | 0.774 | 0.495 | 0.665 | 1 | 0.804 | 0.649 | 0.551 | 0.792 |
| S6 | 0.841 | 0.884 | 0.579 | 0.874 | 0.804 | 1 | 0.888 | 0.739 | 0.941 |
| S7 | 0.831 | 0.883 | 0.654 | 0.859 | 0.649 | 0.888 | 1 | 0.797 | 0.944 |
| S8 | 0.721 | 0.677 | 0.565 | 0.598 | 0.551 | 0.739 | 0.797 | 1 | 0.83 |
| R | 0.934 | 0.916 | 0.731 | 0.847 | 0.792 | 0.941 | 0.944 | 0.83 | 1 |
By above-mentioned table 1 data analysis, calculate greater than 1% by the normalization peak area, 8 kinds of different producing regions red ganoderma samples, maximum fingerprint of alkaloid peak numbers can reach 42 kinds, minimum only is 14 kinds, can be easy to like this differentiate the otherness of the alkaloidal kind of the different contained glossy ganodermas of producing region red ganoderma sample.
In addition, can also distinguish easily the total Fingerprints peak number of lucid ganoderma alkaloids substance, 3.537,3.750,12.979,18.158,19.902,20.156min in above-mentioned 8 glossy ganoderma samples, the total Fingerprints peak number of alkaloid is 6, respectively in retention time:. Calculate by the normalized area integration method, also can differentiate easily, in same retention time section, or to the common characteristic peak, its retention time section, the peak area relative comparison peak-to-peak size of each sample can be differentiated the size of each sample alkaloid, the integral and calculating of total peak area that also can be by each sample and the total peak area of reference substance are differentiated the size of each sample alkaloid.
In addition, by above-mentioned table 2 data analysis, chromatographic peak similarity by each sample is calculated, S1 as can be known, S2, S6, S7 glossy ganoderma sample has more similitude, and its similarity reaches more than 0.916, and the otherness of S3 is maximum, to the raw material quality control of glossy ganoderma sample, the true and false is differentiated the evaluation with scientific meaning like this.
Embodiment 3 adopts different genera glossy ganoderma sample to carry out the alkaloids qualitative analysis
Gather 5 kinds of red sesames of different genera, Han Zhi, purple sesame, black sesame, the Wild ganoderma sample respectively carries out the alkaloids analysis, sampling amount, experiment condition and method of operating are with embodiment 1, obtain 5 kinds of glossy ganoderma sample alkaloids and analyze collection of illustrative plates, then adopt similarity evaluation, choose the red ganoderma sample as reference spectrum peak, and carry out the method for chromatographic peak Supplements, the chromatographic peak of 5 samples mated and automatically generate contrast refer to collection of illustrative plates, see accompanying drawing 3, by coupling and the calculated by peak area of similarity evaluation, Wild ganoderma, black sesame, the more red sesame of Han Zhijun and purple sesame have higher alkaloids substance content. The lucid ganoderma alkaloids substance otherness of different genera is larger, and red sesame, Han Zhi and Hei Zhi are slightly similar, and similarity only reaches about 0.806. Only have the standard finger-print of red sesame finger-print and red ganoderma alkaloids substance to fit like a glove.
Can carry out soon the qualitative discrimination to the height of the alkaloids substance content in the glossy ganoderma of different genera like this.
Claims (9)
1. method for identifying lucid ganoderma alkaloids substance finger-print is characterized in that described method may further comprise the steps:
(a) producing of standard finger-print: 5～10 batches, the every batch crude drug amount of getting equal in quality of ganoderma lucidum fruitbody fragment of getting the different known standard variety sample drying in the place of production is carried out respectively following test: the alcohol reflux that adds 25～35 times of quality of ganoderma lucidum fruitbody fragment of known sample drying extracts, obtain the known sample extract after removing by filter filter residue, the known sample extract is through the decompression distillation desolventizing, get known sample alkaloid medicinal extract liquid, add methanol constant volume and get known sample solution, controlling in the described known sample solution crude drug amount content is/100 milliliters of 2～7 grams, known sample solution is through the organic membrane filtration of 0.45 μ m, get the known sample need testing solution, get the known sample need testing solution and carry out efficient liquid phase chromatographic analysis, obtain known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram; The condition of described high performance liquid chromatography is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, and the detection wavelength is 254nm; Get that the total absworption peak of high-efficient liquid phase chromatogram is Characteristic chromatographic peak in each batch, select among the figure normalization peak area greater than 1% Characteristic chromatographic peak as fingerprint peaks, obtain the standard finger-print of lucid ganoderma alkaloids substance;
(b) the testing sample high-efficient liquid phase chromatogram is produced: the ganoderma lucidum fruitbody fragment of getting the testing sample drying, the alcohol reflux that adds 25～35 times of quality of ganoderma lucidum fruitbody fragment of testing sample drying extracts, obtain the testing sample extract after removing by filter filter residue, the testing sample extract is through the decompression distillation desolventizing, get testing sample alkaloid medicinal extract liquid, add methanol constant volume and get testing sample solution, it is identical to control in the described testing sample solution in the crude drug amount content and step (a) the crude drug amount content of known sample solution, testing sample solution is through the organic membrane filtration of 0.45 μ m, get the testing sample need testing solution, get the testing sample need testing solution and carry out efficient liquid phase chromatographic analysis, the condition of described high performance liquid chromatography is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, the detection wavelength is 254nm, obtains testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram;
Characteristic chromatographic peak contrast in the standard finger-print of the lucid ganoderma alkaloids substance that the testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram that (c) step (b) is obtained and step (a) obtain, testing sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram has feature chromatogram cutting edges of a knife or a sword all in the standard finger-print of lucid ganoderma alkaloids substance, and then testing sample is standard variety.
2. method for identifying lucid ganoderma alkaloids substance finger-print as claimed in claim 1, it is characterized in that in the producing of described method step (a) standard finger-print that the operation that every batch of ganoderma lucidum fruitbody fragment of getting the known standard variety sample drying of equal in quality crude drug amount makes known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram may further comprise the steps:
(1) takes by weighing the ganoderma lucidum fruitbody fragment of known sample drying, add 95wt% ethanol, the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and known sample drying is 25～35: 1, under little reflux state that boils, extracted 1～2 hour, after removing by filter filter residue, get the known sample extract;
(2) the known sample extract is through the decompression distillation desolventizing, get known sample alkaloid medicinal extract liquid, add methanol constant volume and get known sample solution, controlling in the described known sample solution crude drug amount content is/100 milliliters of 2～7 grams, known sample solution gets the known sample need testing solution through the organic membrane filtration of 0.45 μ m;
(3) the known sample need testing solution carries out efficient liquid phase chromatographic analysis, get known sample lucid ganoderma alkaloids substance high-efficient liquid phase chromatogram, described chromatographic condition is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, flow rate of mobile phase is 1mL/min, the detection wavelength is 254nm, sample size 10 μ L, be 40min analysis time, the gradient elution program is: 0～8min, the 1vol% aqueous acetic acid of 10vol% and the acetonitrile of 90vol%; 8～10min, the 1vol% aqueous acetic acid of 15vol% and the acetonitrile of 85vol%; 10～15min, the 1vol% aqueous acetic acid of 30vol% and the acetonitrile of 70vol%; 15～20min, the 1vol% aqueous acetic acid of 40vol% and the acetonitrile of 60vol%; 20～30min, the 1vol% aqueous acetic acid of 45vol% and the acetonitrile of 55vol%; 30～40min, the 1vol% aqueous acetic acid of 50vol% and the acetonitrile of 50vol%.
3. method for identifying lucid ganoderma alkaloids substance finger-print as claimed in claim 1 is characterized in that the operation that the testing sample high-efficient liquid phase chromatogram is produced in the described method step (b) may further comprise the steps:
(1) takes by weighing the ganoderma lucidum fruitbody fragment of testing sample drying, add 95wt% ethanol, the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and testing sample drying is 25～35: 1, under little reflux state that boils, extracted 1～2 hour, after removing by filter filter residue, get the testing sample extract;
(2) the testing sample extract is through the decompression distillation desolventizing, get testing sample alkaloid medicinal extract liquid, add methanol constant volume and get testing sample solution, it is identical to control in the described testing sample solution in the crude drug amount content and step (a) the crude drug amount content of known sample solution, testing sample solution gets the testing sample need testing solution through the organic membrane filtration of 0.45 μ m;
(3) the testing sample need testing solution carries out efficient liquid phase chromatographic analysis, get high-efficient liquid phase chromatogram, described chromatographic condition is: the chromatographic column take octadecylsilane chemically bonded silica as filler, carry out gradient elution take own nitrile, 1vol% aqueous acetic acid as mobile phase, flow rate of mobile phase is 1mL/min, and the detection wavelength is 254nm, sample size 10 μ L, be 40min analysis time, and the gradient elution program is: 0～8min, the 1vol% aqueous acetic acid of 10vol% and the acetonitrile of 90vol%; 8～10min, the 1vol% aqueous acetic acid of 15vol% and the acetonitrile of 85vol%; 10～15min, the 1vol% aqueous acetic acid of 30vol% and the acetonitrile of 70vol%; 15～20min, the 1vol% aqueous acetic acid of 40vol% and the acetonitrile of 60vol%; 20～30min, the 1vol% aqueous acetic acid of 45vol% and the acetonitrile of 55vol%; 30～40min, the 1vol% aqueous acetic acid of 50vol% and the acetonitrile of 50vol%.
4. method as claimed in claim 2 is characterized in that in the described step (1) described being extracted as: under little reflux state that boils, extracted 1 hour, remove by filter filter residue after, get the known sample extract.
5. method as claimed in claim 3 is characterized in that in the described step (1) described being extracted as: under little reflux state that boils, extracted 1 hour, remove by filter filter residue after, get the testing sample extract.
6. method as claimed in claim 2, it is characterized in that in the described step (1) described being extracted as: power is under the microwave condition of 800W, microwave refluxing extraction 17min, remove by filter filter residue after, get the known sample extract.
7. method as claimed in claim 3, it is characterized in that in the described step (1) described being extracted as: power is under the microwave condition of 800W, microwave refluxing extraction 17min, remove by filter filter residue after, get the testing sample extract.
8. method as claimed in claim 2 is characterized in that in the described step (1), the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and known sample drying is 33: 1.
9. method as claimed in claim 3 is characterized in that in the described step (1), the mass ratio of the ganoderma lucidum fruitbody fragment of described 95wt% ethanol and testing sample drying is 33: 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101547749A CN101732364B (en) | 2009-12-07 | 2009-12-07 | Method for identifying lucid ganoderma alkaloids substance finger-print |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101547749A CN101732364B (en) | 2009-12-07 | 2009-12-07 | Method for identifying lucid ganoderma alkaloids substance finger-print |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101732364A true CN101732364A (en) | 2010-06-16 |
| CN101732364B CN101732364B (en) | 2012-05-23 |
Family
ID=42456732
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101547749A Expired - Fee Related CN101732364B (en) | 2009-12-07 | 2009-12-07 | Method for identifying lucid ganoderma alkaloids substance finger-print |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101732364B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735772A (en) * | 2012-07-02 | 2012-10-17 | 福建省药品检验所 | Quality controlling method of ganoderma aqueous extract |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107782822A (en) * | 2017-10-18 | 2018-03-09 | 中国农业科学院特产研究所 | The assay method of ganoderic acid content in a kind of ganoderma lucidum |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100362345C (en) * | 2005-07-21 | 2008-01-16 | 广州汉方现代中药研究开发有限公司 | Establishment of lucid ganderma spore and lucid ganderma spore oil finger print atlas and standard finger print atlas |
| CN101435801A (en) * | 2007-11-13 | 2009-05-20 | 南京中科集团股份有限公司 | Fingerprint pattern identifying method of ganoderma lucidum spore oil |
-
2009
- 2009-12-07 CN CN2009101547749A patent/CN101732364B/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735772A (en) * | 2012-07-02 | 2012-10-17 | 福建省药品检验所 | Quality controlling method of ganoderma aqueous extract |
| CN102735772B (en) * | 2012-07-02 | 2014-05-28 | 福建省药品检验所 | Quality controlling method of ganoderma aqueous extract |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101732364B (en) | 2012-05-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yu et al. | Adulteration and cultivation region identification of American ginseng using HPLC coupled with multivariate analysis | |
| CN1895617A (en) | Kidney-warming and heart-nourishing Chinese-medicinal preparation, its making method and quality control | |
| CN101229219A (en) | Technology of processing astragalus root decoction pieces | |
| CN102028859B (en) | Detection method for Chinese medicinal preparation for treating asthma | |
| CN108578571A (en) | A kind of concocting method that can directly take orally sealwort decoction piece | |
| CN101695520B (en) | Preparation method of medicament for treating diabetes | |
| CN103558329B (en) | Method for establishing fingerprint of leech and standard fingerprint of leech | |
| CN101732364B (en) | Method for identifying lucid ganoderma alkaloids substance finger-print | |
| CN114200046A (en) | Construction method and detection method of Juanbi decoction composition fingerprint spectrum and application | |
| CN113189245B (en) | Platycodon grandiflorum soup material standard and establishing method thereof | |
| CN113376273B (en) | Detection method of HPLC (high performance liquid chromatography) characteristic spectrum of Qingxin lotus seed drink and application of characteristic spectrum | |
| CN101991669B (en) | Quality control method of medicine | |
| CN101181328A (en) | Method for producing ginseng fruit and ginseng flower stalk with high content of ginsenoside | |
| CN115778997B (en) | Preparation method and quality detection method of astragalus root and burdock granules | |
| CN103202873B (en) | Medicine composition used for enhancing human immunity | |
| CN102028883B (en) | Standard fingerprint spectrum of Chinese medicinal composition, and measurement method and application thereof | |
| CN106124668A (en) | A kind of fingerprint atlas detection method of Yixinshu tablet | |
| CN116298041A (en) | Method for measuring HPLC fingerprint of Guanxintai pill | |
| CN106018647B (en) | A kind of method for setting up sunflower disk HPLC standard finger-prints | |
| CN113759023A (en) | Preparation process and evaluation method of angelica sinensis Sini decoction | |
| CN102302542A (en) | Chinese medicinal oral liquid for treating cancers | |
| CN106918657A (en) | A kind of method for building up of the finger-print of Chinese medicinal composition preparation | |
| CN112684091A (en) | HPLC-DAD-based six-ingredient rehmannia pill formula analysis and quality analysis method | |
| CN1879849A (en) | Compound capsule with pseudo-ginseng and Chinese fanpalm seed, its preparation process and quality control method | |
| CN115887598B (en) | Preparation method of compound preparation of channel warming soup |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120523 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |