CN107782822A - The assay method of ganoderic acid content in a kind of ganoderma lucidum - Google Patents

The assay method of ganoderic acid content in a kind of ganoderma lucidum Download PDF

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Publication number
CN107782822A
CN107782822A CN201710971176.5A CN201710971176A CN107782822A CN 107782822 A CN107782822 A CN 107782822A CN 201710971176 A CN201710971176 A CN 201710971176A CN 107782822 A CN107782822 A CN 107782822A
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ganoderma lucidum
acid content
ganoderic acid
assay method
ganoderic
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逄世峰
闫梅霞
崔丽丽
王佳
张瑞
王英平
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Institute Special Animal and Plant Sciences CAAS
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Institute Special Animal and Plant Sciences CAAS
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Pathology (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The invention provides a kind of assay method of ganoderic acid content in ganoderma lucidum, comprise the following steps:(A) the fructification drying, crushing plus ethanol, ultrasonic extraction of ganoderma lucidum are obtained into test liquid;(B) test liquid is subjected to efficient liquid phase chromatographic analysis, mobile phase is acetonitrile phosphate aqueous solution, and flow control is between 0.3 0.5ml/min, 30 40 DEG C of column temperature, between the 260nm of Detection wavelength 250;(C) injection test liquid carries out analysis measure, you can.The assay method specificity of ganoderic acid content is strong in the ganoderma lucidum of the embodiment of the present invention, favorable reproducibility, used time is short, and the detection method has filled up the correlation technique blank being measured using high performance liquid chromatography to ganoderic acid content in ganoderma lucidum, form the detection architecture of standard of comparison, suitable for wide popularization and application, highly use for reference.

Description

The assay method of ganoderic acid content in a kind of ganoderma lucidum
Technical field
The present invention relates to the measure field of ganoderic acid in ganoderma lucidum, in particular to ganoderic acid content in a kind of ganoderma lucidum Assay method.
Background technology
Ganoderma lucidum (scientific name:Ganoderma Lucidum Karst) it is also known as polyporus lucidus, celestial grass, seocho, profile is in umbrella, bacterium Lid kidney shape, semicircle or subcircular, Basidiomycetes Polyporaceae Ganoderma fungi, for the fructification of On Polyporaceae ganoderma lucidum. Ganoderma lucidum is generally distributed in China, Guizhou, Heilungkiang, Jilin, Shandong, Hunan, Huoshan, Jiangxi, Fujian, Guangdong, Guangxi, river There is part yield in the provinces such as north.Wherein northeast ganoderma lucidum planting base is with Jilin Province Jiaohe City yellow pine pasture town and Dunhua of Jilin Province city More concentrate in yellow mud river town.
Ganoderma lucidum is medicinal in China's history of existing more than 2000 years, is considered as strengthening by means of tonics by successive dynasties medicine man, strengthens the body resistance to consolidate the constitution Magical treasure.2000, it is known that Ganoderma (Ganoderma) fungi about more than 100 is planted, and distribution is most wide for red sesame (G.lucidum) it is secondly, purple sesame (G.japonicum), also artist's conk (G.applanatum), Ganoderma tsugae (G.tsugae) With the equal hyoscine such as book tree sesame (G.capense).By a large amount of clinical researches, ganoderma lucidum is to neurasthenia, hyperlipidemia, coronary heart disease The degree of having nothing in common with each other such as angina pectoris, arrhythmia cordis, Keshan disease, plateau uncomfortable diseases, hepatitis, Hemorrhagic fever, indigestion, tracheitis Curative effect.
Ganoderma lucidum has an invigorating qi for tranquilization, relieving cough and asthma and other effects.Research shows, the chemical composition of ganoderma lucidum has a variety of, and master contains ammonia Base acid, polypeptide, protein, fungal lysozyme (fungal lysozyme), and carbohydrate (reduced sugar and polysaccharide), ergosterol, Triterpenes, coumarin glucoside, volatile oil, stearic acid, benzoic acid, alkaloid, vitamin B2 and C etc.;Spore is also containing mannitol, marine alga Sugared (trehalose), wherein triterpene compound is the main active substances in ganoderma lucidum, has extensive pharmacological activity, such as anti- Tumour, anti-HIV-1 type virus, protect liver, removing toxic substances and improvement memory etc., therefore, are measured to the ganoderic acid content in ganoderma lucidum Extremely it is necessary.
In the prior art, assay is carried out to the ganoderic acid in ganoderma lucidum mainly using Ultraviolet spectrophotometry, but It is that this method specificity is not strong, testing result is inaccurate, to the characterization and evaluation of the follow-up medical value of ganoderma lucidum, and marketing Dynamics can all have a certain impact.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide a kind of assay method of ganoderic acid content in ganoderma lucidum, the assay method is special pin A kind of high-efficiency liquid chromatography method for detecting being designed to the measure of the ganoderic acid content in ganoderma lucidum, specificity is strong, reappearance Good, the used time is short, and the detection method has filled up the correlation being measured using high performance liquid chromatography to ganoderic acid content in ganoderma lucidum Technological gap, the detection architecture of standard of comparison is formd, suitable for wide popularization and application, highly used for reference.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme:
It is main to include following step the embodiments of the invention provide a kind of assay method for being suitable to ganoderic acid content in ganoderma lucidum Suddenly:
(A) the fructification drying, crushing plus ethanol, ultrasonic extraction of ganoderma lucidum are obtained into test liquid;
(B) test liquid is subjected to efficient liquid phase chromatographic analysis, mobile phase is acetonitrile-phosphate aqueous solution, and flow control exists Between 0.3-0.5ml/min, 30-40 DEG C of column temperature, between Detection wavelength 250-260nm;
(C) injection test liquid carries out analysis measure, you can.
Ganoderma lucidum is medicinal in China's history of existing more than 2000 years, is considered as strengthening by means of tonics by successive dynasties medicine man, strengthens the body resistance to consolidate the constitution Magical treasure.It is relieving cough and asthma and other effects with invigorating qi for tranquilization.Research shows, the chemical composition of ganoderma lucidum has a variety of, and master contains amino Acid, polypeptide, protein, fungal lysozyme (fungallysozyme), and carbohydrate (reduced sugar and polysaccharide), ergosterol, three Terpene, coumarin glucoside, volatile oil, stearic acid, benzoic acid, alkaloid, vitamin B2 and C etc.;Spore is also containing mannitol, trehalose (trehalose), wherein triterpene compound is the main active substances in ganoderma lucidum, has extensive pharmacological activity, such as anti-swollen Knurl, anti-HIV-1 type virus, protect liver, removing toxic substances and improvement memory etc., therefore, the ganoderic acid content in ganoderma lucidum, which is measured, is Extremely it is necessary.
In the prior art, assay is carried out to the ganoderic acid in ganoderma lucidum mainly using Ultraviolet spectrophotometry, but It is that this method specificity is not strong, testing result is inaccurate, to the characterization and evaluation of the follow-up medical value of ganoderma lucidum, and marketing Dynamics can all have a certain impact.
The present invention is in order to solve the above technical problems, provide a kind of measure side of the ganoderic acid content suitable for ganoderma lucidum Method, has abandoned the conventional traditional detection method using Ultraviolet spectrophotometry, and this method has that operation is simple, repeatability It is good, the degree of accuracy is high, measure relative error (RSD) is less than 5%, can extensive use, filled up using high performance liquid chromatography to determine The relevant art blank of ganoderic acid content in ganoderma lucidum, have and be widely popularized value, expanded the applicable surface of the medical value of ganoderma lucidum, Also the marketing degree of ganoderma lucidum is more favorable for, and then certain economic benefit can be created.
In specific operating procedure (A) of the invention, before using efficient liquid phase chromatographic analysis, first ganoderma lucidum medicinal material is entered Row a series of purification operation, that is, dry, crush plus ethanol, to obtain test liquid after ultrasonic extraction.
Wherein, the temperature of drying is preferably controlled in 30-50 DEG C, and more excellent is 45 DEG C, and being typically dried to constant weight can meet It is required that can evaporate the moisture on ganoderma lucidum surface as far as possible totally under these conditions, follow-up testing result is not interfered with.
Preferably, after the fructification drying of ganoderma lucidum, for the mesh Task-size Controlling of crushing more than 60 mesh, more excellent is 70-100 mesh, In addition can also be 80 mesh, 90 mesh, 120 mesh etc..Ultrasonic extraction mistake will be subsequently being carried out after ganoderma lucidum attrition grinding powdering The extraction of ganoderic acid is more beneficial in journey.
Preferably, ganoderma lucidum fruitbody is extracted as solvent using ethanol, every gram of ganoderma lucidum fruitbody addition ethanol 20-30ml, the volume of more excellent addition ethanol is 25-28ml, and the concentration general control of ethanol is between 70-90wt%.
In order that obtain ganoderma lucidum fruitbody fully can be well mixed with ethanol, to be carried out at corresponding ultrasound after adding ethanol Reason operation extraction, in more than 500w, FREQUENCY CONTROL more preferably exists for ultrasonic power control in more than 40kHz for ultrasonic power control Between 600-700w, FREQUENCY CONTROL the restriction of appropriate frequency and power, extracts obtained ganoderic acid between 45-50kHz Purity is higher.
Also, it is ultrasonically treated 1-2 times, time control per treatment is in more than 30min.
Preferably, extract and twice afterwards merge extract solution repeatedly, finally carried out using 0.22 μm of PTFE filter membrane Filter, to ensure the homogeneity of test liquid, fully remove contained impurity.
Preferably, the temperature control of extraction is between 20-30 DEG C.
The method of a most appropriate ganoderic acid extraction is found out by the design of experimental condition, by each operating parameter Control than in convenient scope, it is easy to operate, meet the requirement of ganoderic acid extracting method.
In the step (B) of the present invention, the actual conditions of efficient liquid phase chromatographic analysis is:Mobile phase is that acetonitrile-phosphoric acid is water-soluble Liquid, flow control is between 0.3-0.5ml/min, 30-40 DEG C of column temperature, between Detection wavelength 250-260nm.Wherein, chromatographic column It is preferably selected ACQUITY UPLC HSS T3 (2.1 × 50mm, 1.7 μm).
Preferably, selected mobile phase is acetonitrile-phosphate aqueous solution, gradient elution:0-13min, 19%-29.4% second Nitrile;13-16min, 29.4%-45% acetonitrile.
Preferably, 30-40 DEG C of column temperature, more excellent is 35 DEG C.
Preferably, between Detection wavelength 250-260nm, more excellent is 258nm.
Preferably, in acetonitrile-phosphate aqueous solution, for phosphonic acids volumetric concentration between 0.02-0.05%, more excellent is 0.03%.
General sample size be 2 μ L, accurate respectively during practical operation to draw reference substance solution and each 2 μ L of need testing solution, is noted Enter Ultra Performance Liquid Chromatography instrument measure, peak area quantification qualitative with main peak retention time.
The assay method of the high performance liquid chromatography of the present invention can determine red sesame acid C in ganoderma lucidum, red sesame acid N, Lanost-8-en-26-oic acid,3,7,12-trihydroxy-11,15,23-trioxo-,(3BETA,7BETA,12BETA)-, spirit Sesame olefin(e) acid B, ganoderic acid B, red sesame acid E, red sesame acid B, desacetoxy ganoderic acid F, ganoderma lucidum olefin(e) acid E, ganoderic acid A, lucidenic acid A, ganoderma lucidum Sour D, red sesame acid D, ganoderic acid F contents, the scope of application are very extensive.
Compared with prior art, beneficial effects of the present invention are:
(1) the invention provides a kind of assay method of ganoderic acid content in ganoderma lucidum, the assay method to be specific to spirit A kind of high-efficiency liquid chromatography method for detecting that the measure of ganoderic acid content in sesame is designed, specificity is strong, favorable reproducibility, uses When it is short, and to have filled up the correlation technique being measured to ganoderic acid content in ganoderma lucidum using high performance liquid chromatography empty for the detection method In vain, the detection architecture of standard of comparison is formd, suitable for wide popularization and application, is highly used for reference;
(2) present invention passes through to drying, crushing and be defined the step of ultrasonic extraction so that whole extraction step work Sequence is simple, with short production cycle, and has found out the method that a most appropriate ganoderic acid extracts by the design of experimental condition, Each operating parameter is controlled than in convenient scope, it is easy to operate, meet the requirement of ganoderic acid extracting method.
Brief description of the drawings
, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical scheme of the prior art The required accompanying drawing used is briefly described in embodiment or description of the prior art, it should be apparent that, in describing below Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before creative work is not paid Put, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 is the high-efficient liquid phase chromatogram of the ganoderma lucidum standard solution of the embodiment of the present invention 1;
Fig. 2 is the high-efficient liquid phase chromatogram of the ganoderma lucidum need testing solution of the embodiment of the present invention 1;
Reference:
1:Red sesame acid C;2:Red sesame acid N;3:Lanost-8-en-26-oic acid,3,7,12-trihydroxy-11,15,23-trioxo-,(3BETA,7BETA,12BETA)-;4:Ganoderma lucidum olefin(e) acid B;
5:Ganoderic acid B;6:Desacetoxy ganoderic acid F;7:Ganoderic acid A;8:Lucidenic acid A;
9:Ganoderic acid D;10:Red sesame acid D;11:Ganoderic acid F.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is The conventional products that can be obtained by commercially available purchase.
Embodiment 1
The specific assay method of ganoderic acid content is as follows in ganoderma lucidum:
Ganoderma lucidum fruitbody is dried, crushed, after crossing 60 mesh sieves, takes 2.0g sample powder, it is accurately weighed, add ethanol 40mL, room temperature ultrasound (power 500w, frequency 40kHz) extraction 20min, filtering, is extracted 2 times, merges extract solution to 100mL capacity In bottle, scale is settled to, is shaken up, stood, Aspirate supernatant, cross 0.22 μm of PTFE filter membrane, treat that sample introduction is analyzed.
Chromatographic condition:
ACQUITY UPLC HSS T3 chromatographic columns (2.1 × 50mm, 1.7 μm), mobile phase:The phosphoric acid of acetonitrile -0.03% is water-soluble Liquid (gradient elution:0-13min, 19%-29.4% acetonitrile;13~16min, 29.4%-45% acetonitrile), flow velocity 0.5ml/min, 35 DEG C of column temperature, sample size 2 μ l, Detection wavelength 250nm.
The high-efficient liquid phase chromatogram of specific standards product and need testing solution is as shown in accompanying drawing 1-2.
Specification Curve of Increasing:
By mixed reference substance solution methanol dilution, a series of hybrid standard product solution of concentration is made, is implanted sequentially height Effect liquid phase chromatogram instrument, peak area is determined by above-mentioned chromatographic condition, using peak area as ordinate, concentration is abscissa, draws standard Curve, calculate regression equation.
Sample determines:
The μ l of need testing solution 2 are drawn, are analyzed by above-mentioned chromatographic condition sample introduction, peak area is brought into regression equation, calculates and supply Ganoderma lucidum acid concentration in test sample solution.
The concentration of ganoderic acid in C-need testing solution, unit are every milliliter of milligram (mg/ml);
V-need testing solution constant volume, unit are milliliter (ml);
M-sample mass, unit are gram (g);
Error:The absolute difference of the measurement result independent twice obtained under repeat condition is no more than arithmetic mean of instantaneous value 5%.
Embodiment 2
The specific assay method of ganoderic acid content is as follows in ganoderma lucidum:
By 30 DEG C of drying of ganoderma lucidum fruitbody, crush, after crossing 70 mesh sieves, take 2.0g sample powder, it is accurately weighed, add 70wt% ethanol 60mL, 30 DEG C of extractions of room temperature ultrasound (power 600w, frequency 45kHz, ultrasound 2 times, each ultrasonic 30min), Filtering, merge extract solution into 100mL volumetric flasks, be settled to scale, shake up, stand, Aspirate supernatant, cross 0.22 μm of PTFE Filter membrane, treat that sample introduction is analyzed.
Chromatographic condition:
ACQUITY UPLC HSS T3 chromatographic columns (2.1 × 50mm, 1.7 μm), mobile phase:The phosphoric acid of acetonitrile -0.02% is water-soluble Liquid (gradient elution:0-13min, 19%-29.4% acetonitrile;13~16min, 29.4%-45% acetonitrile), flow velocity 0.3ml/min, 35 DEG C of column temperature, sample size 2 μ l, Detection wavelength 258nm.
Specification Curve of Increasing:
By mixed reference substance solution methanol dilution, a series of hybrid standard product solution of concentration is made, is implanted sequentially height Effect liquid phase chromatogram instrument, peak area is determined by above-mentioned chromatographic condition, using peak area as ordinate, concentration is abscissa, draws standard Curve, calculate regression equation.
Sample determines:
The μ l of need testing solution 2 are drawn, are analyzed by above-mentioned chromatographic condition sample introduction, peak area is brought into regression equation, calculates and supply Ganoderma lucidum acid concentration in test sample solution.
The concentration of ganoderic acid in C-need testing solution, unit are every milliliter of milligram (mg/ml);
V-need testing solution constant volume, unit are milliliter (ml);
M-sample mass, unit are gram (g);
Error:The absolute difference of the measurement result independent twice obtained under repeat condition is no more than arithmetic mean of instantaneous value 5%.
Embodiment 3
The specific assay method of ganoderic acid content is as follows in ganoderma lucidum:
By 50 DEG C of drying of ganoderma lucidum fruitbody, crush, after crossing 100 mesh sieves, take 2.0g sample powder, it is accurately weighed, add 90wt% ethanol 50mL, 20 DEG C of extractions of room temperature ultrasound (power 600w, frequency 50kHz, ultrasound 1 time, ultrasonic 35min), filtering, Extract solution is put into 100mL volumetric flasks, is settled to scale, is shaken up, is stood, Aspirate supernatant, crosses 0.22 μm of PTFE filter membrane, Treat that sample introduction is analyzed.
Chromatographic condition:
ACQUITY UPLC HSS T3 chromatographic columns (2.1 × 50mm, 1.7 μm), mobile phase:The phosphoric acid of acetonitrile -0.02% is water-soluble Liquid (gradient elution:0-13min, 19%-29.4% acetonitrile;13~16min, 29.4%-45% acetonitrile), flow velocity 0.4ml/min, 40 DEG C of column temperature, sample size 2 μ l, Detection wavelength 250nm.
Specification Curve of Increasing:
By mixed reference substance solution methanol dilution, a series of hybrid standard product solution of concentration is made, is implanted sequentially height Effect liquid phase chromatogram instrument, peak area is determined by above-mentioned chromatographic condition, using peak area as ordinate, concentration is abscissa, draws standard Curve, calculate regression equation.
Sample determines:
The μ l of need testing solution 2 are drawn, are analyzed by above-mentioned chromatographic condition sample introduction, peak area is brought into regression equation, calculates and supply Ganoderma lucidum acid concentration in test sample solution.
The concentration of ganoderic acid in C-need testing solution, unit are every milliliter of milligram (mg/ml);
V-need testing solution constant volume, unit are milliliter (ml);
M-sample mass, unit are gram (g);
Error:The absolute difference of the measurement result independent twice obtained under repeat condition is no more than arithmetic mean of instantaneous value 5%.
Embodiment 4
The specific assay method of ganoderic acid content is as follows in ganoderma lucidum:
By 45 DEG C of drying of ganoderma lucidum fruitbody, crush, after crossing 100 mesh sieves, take 2.0g sample powder, it is accurately weighed, add 75wt% ethanol 50mL, 25 DEG C of extractions of room temperature ultrasound (power 600w, frequency 48kHz, ultrasound 2 times, each ultrasonic 30min), Filtering, merge extract solution into 100mL volumetric flasks, be settled to scale, shake up, stand, Aspirate supernatant, cross 0.22 μm of PTFE Filter membrane, treat that sample introduction is analyzed.
Chromatographic condition:
ACQUITY UPLC HSS T3 chromatographic columns (2.1 × 50mm, 1.7 μm), mobile phase:The phosphoric acid of acetonitrile -0.05% is water-soluble Liquid (gradient elution:0-13min, 19%-29.4% acetonitrile;13~16min, 29.4%-45% acetonitrile), flow velocity 0.4ml/min, 35 DEG C of column temperature, sample size 2 μ l, Detection wavelength 260nm.
Subsequent step is consistent with embodiment 2.
Error:The absolute difference of the measurement result independent twice obtained under repeat condition is no more than arithmetic mean of instantaneous value 5%.
Comparative example 1
By using the Ultraviolet spectrophotometry of embodiment 1 in the patent of Application No. 201510949610.0 in ganoderma lucidum After the content of ganoderic acid is measured, the absolute difference of the measurement result independent twice obtained under repeat condition is arithmetic average The 15% of value.
Compared with prior art, beneficial effects of the present invention are:
(1) the invention provides a kind of assay method of ganoderic acid content in ganoderma lucidum, the assay method to be specific to spirit A kind of high-efficiency liquid chromatography method for detecting that the measure of ganoderic acid content in sesame is designed, specificity is strong, favorable reproducibility, uses When it is short, and to have filled up the correlation technique being measured to ganoderic acid content in ganoderma lucidum using high performance liquid chromatography empty for the detection method In vain, the detection architecture of standard of comparison is formd, suitable for wide popularization and application, is highly used for reference;
(2) present invention passes through to drying, crushing and be defined the step of ultrasonic extraction so that whole extraction step work Sequence is simple, with short production cycle, and has found out the method that a most appropriate ganoderic acid extracts by the design of experimental condition, Each operating parameter is controlled than in convenient scope, it is easy to operate, meet the requirement of ganoderic acid extracting method.
Although illustrate and describing the present invention with specific embodiment, but will be appreciated that without departing substantially from the present invention's Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims Including belonging to all such changes and modifications in the scope of the invention.

Claims (10)

1. the assay method of ganoderic acid content in a kind of ganoderma lucidum, it is characterised in that mainly comprise the following steps:
(A) the fructification drying, crushing plus ethanol, ultrasonic extraction of ganoderma lucidum are obtained into test liquid;
(B) test liquid is subjected to efficient liquid phase chromatographic analysis, mobile phase is acetonitrile-phosphate aqueous solution, and flow control is in 0.3- Between 0.5ml/min, 30-40 DEG C of column temperature, between Detection wavelength 250-260nm;
(C) injection test liquid carries out analysis measure, you can.
2. the assay method of ganoderic acid content in ganoderma lucidum according to claim 1, it is characterised in that in the step (A), After the fructification drying of ganoderma lucidum, for the mesh Task-size Controlling of crushing more than 60 mesh, more excellent is 70-100 mesh;
Preferably, the temperature of drying is 30-50 DEG C, and more excellent is 45 DEG C.
3. the assay method of ganoderic acid content in ganoderma lucidum according to claim 1, it is characterised in that in the step (A), Every gram of ganoderma lucidum fruitbody addition ethanol 20-30ml, the volume of more excellent addition ethanol is 25-28ml;
Preferably, the concentration of ethanol is controlled between 70-90wt%.
4. the assay method of ganoderic acid content in ganoderma lucidum according to claim 1, it is characterised in that in the step (A), Supersound process extraction is carried out after addition ethanol, Power Control is in more than 500w;
Preferably, ultrasonic FREQUENCY CONTROL is more excellent between 45-50kHz in more than 40kHz.
5. the assay method of ganoderic acid content in ganoderma lucidum according to claim 4, it is characterised in that in the step (A), It is ultrasonically treated 1-2 times, time control per treatment is in more than 30min.
6. the assay method of ganoderic acid content in ganoderma lucidum according to claim 1, it is characterised in that in the step (A), Merge extract solution and filtered using 0.22 μm of PTFE filter membrane.
7. the assay method of ganoderic acid content in ganoderma lucidum according to claim 6, it is characterised in that the temperature control of extraction Between 20-30 DEG C.
8. the assay method of ganoderic acid content in the ganoderma lucidum according to claim any one of 1-6, it is characterised in that the step Suddenly in (B), the chromatographic column of efficient liquid phase chromatographic analysis is ACQUITY UPLC HSS T3.
9. the assay method of ganoderic acid content in the ganoderma lucidum according to claim any one of 1-6, it is characterised in that the step Suddenly in (B), 30-40 DEG C of column temperature, more excellent is 35 DEG C.
10. the assay method of ganoderic acid content in the ganoderma lucidum according to claim any one of 1-6, it is characterised in that described In step (B), between Detection wavelength 250-260nm, more excellent is 258nm;
Preferably, in acetonitrile-phosphate aqueous solution, for phosphonic acids volumetric concentration between 0.02-0.05%, more excellent is 0.03%.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108802255A (en) * 2018-06-15 2018-11-13 福建仙芝楼生物科技有限公司 The method for measuring ganoderic acid A and Ganoderma lucidum triterpenes components content in compound preparation
CN111562251A (en) * 2020-05-26 2020-08-21 河北省食品检验研究院 Method for rapidly detecting ganoderic acid B in ganoderma lucidum spore powder
CN112326853A (en) * 2020-11-06 2021-02-05 上海市农业科学院 Method for simultaneously detecting 25 triterpene compounds in ganoderma lucidum fruiting body
CN112345681A (en) * 2020-11-06 2021-02-09 上海市农业科学院 Method for simultaneously detecting eight ganoderic acids in ganoderma lucidum mycelia
CN113109556A (en) * 2021-04-19 2021-07-13 河北农业大学 Detection method of ganoderic acid A

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006022017A (en) * 2004-07-06 2006-01-26 Univ Nihon Carcinogenesis prophylactic agent
CN101671383A (en) * 2009-10-19 2010-03-17 上海交通大学 Monomeric 7-ethyoxyl ganoderic acid O and separating and purifying method of monomeric 7-ethyoxyl ganoderic acid T thereof
CN101732364B (en) * 2009-12-07 2012-05-23 浙江工业大学 Method for identifying lucid ganoderma alkaloids substance finger-print
CN102721764A (en) * 2012-07-02 2012-10-10 福建省药品检验所 Method for controlling quality of ganoderma lucidum alcohol extract
TWI381844B (en) * 2010-01-12 2013-01-11 Yung Kien Ind Corp A method for isolating ganoderic acids from ganoderma growth bags
CN105092765A (en) * 2015-09-15 2015-11-25 上海市农业科学院 Detection method for effective components in ganoderma lucidum spore powder raw material
CN106338563A (en) * 2016-09-13 2017-01-18 广州神农生物技术有限公司 Quality evaluation method for ganoderma lucidum medicinal material

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006022017A (en) * 2004-07-06 2006-01-26 Univ Nihon Carcinogenesis prophylactic agent
CN101671383A (en) * 2009-10-19 2010-03-17 上海交通大学 Monomeric 7-ethyoxyl ganoderic acid O and separating and purifying method of monomeric 7-ethyoxyl ganoderic acid T thereof
CN101732364B (en) * 2009-12-07 2012-05-23 浙江工业大学 Method for identifying lucid ganoderma alkaloids substance finger-print
TWI381844B (en) * 2010-01-12 2013-01-11 Yung Kien Ind Corp A method for isolating ganoderic acids from ganoderma growth bags
CN102721764A (en) * 2012-07-02 2012-10-10 福建省药品检验所 Method for controlling quality of ganoderma lucidum alcohol extract
CN105092765A (en) * 2015-09-15 2015-11-25 上海市农业科学院 Detection method for effective components in ganoderma lucidum spore powder raw material
CN106338563A (en) * 2016-09-13 2017-01-18 广州神农生物技术有限公司 Quality evaluation method for ganoderma lucidum medicinal material

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
FLORIAN HENNICKE等: "Distinguishing commercially grown Ganoderma lucidum from Ganoderma lingzhi from Europe and East Asia on the basis of morphology, molecular phylogeny, and triterpenic acid prfiles", 《PHYTOCHEMISTRY》 *
JUAN DA等: "Comparison of two officinal Chinese pharmacopoeia species of Ganoderma basedon chemical research with multiple technologies and chemometrics analysis", 《JOURNAL OF CHROMATOGRAPHY A》 *
MIN YANG等: "Analysis of Triterpenoids in Ganoderma lucidum Using Liquid Chromatography Coupled with Electrospray Ionization Mass Spectrometry", 《J AM SOC MASS SPECTROM》 *
ZHOU YAN等: "Fast analysis of triterpenoids in Ganoderma lucidum spores by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry", 《BIOMEDICLA CHROMATOGRAPHY》 *
刘良琴等: "超高效液相色谱-串联质谱法测定灵芝红茶中灵芝酸A和灵芝酸B", 《理化检验-化学分册》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108802255A (en) * 2018-06-15 2018-11-13 福建仙芝楼生物科技有限公司 The method for measuring ganoderic acid A and Ganoderma lucidum triterpenes components content in compound preparation
CN111562251A (en) * 2020-05-26 2020-08-21 河北省食品检验研究院 Method for rapidly detecting ganoderic acid B in ganoderma lucidum spore powder
CN112326853A (en) * 2020-11-06 2021-02-05 上海市农业科学院 Method for simultaneously detecting 25 triterpene compounds in ganoderma lucidum fruiting body
CN112345681A (en) * 2020-11-06 2021-02-09 上海市农业科学院 Method for simultaneously detecting eight ganoderic acids in ganoderma lucidum mycelia
CN112345681B (en) * 2020-11-06 2023-02-03 上海市农业科学院 Method for simultaneously detecting eight ganoderic acids in ganoderma lucidum mycelia
CN113109556A (en) * 2021-04-19 2021-07-13 河北农业大学 Detection method of ganoderic acid A

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Application publication date: 20180309