CN103948651A

CN103948651A - Application of freezing technology in drying of lucid ganoderma

Info

Publication number: CN103948651A
Application number: CN201410204837.8A
Authority: CN
Inventors: 卢宁; 郭庆华; 冯立娟; 丁桂丽; 孙志芳
Original assignee: Jinan Kangzhong Pharmaceutical Research and Development Co Ltd
Current assignee: Jinan Kangzhong Pharmaceutical Research and Development Co Ltd
Priority date: 2014-05-15
Filing date: 2014-05-15
Publication date: 2014-07-30

Abstract

The invention relates to application of the freezing technology in drying of lucid ganoderma. According to the preparation, fresh lucid ganoderma is picked and then treated by freezing, piling for 2 to 5 days and drying; the prepared lucid ganoderma is high in content of active ingredient, namely, polysaccharide, and also high in anti-cancer activity.

Description

The application of Refrigeration Technique in Ganoderma is dry

Technical field

The present invention relates to medicine method for making, the particularly application of Refrigeration Technique in Ganoderma is dry, belongs to medical technical field.

Background technology

Ganoderma is On Polyporaceae Ganoderma lucidum (Leyss. Ex Fr.) Karst. Ganodermalucidum (Leyss.ExFr.) Karst. or Ganoderma GanodermasinenseZhao, the dry sporophore of XuetZhang.There is invigorating QI and tranquilization, the effect of relieving cough and asthma, for dizzy sleeplessness, shortness of breath and palpitation, asthenia cough with asthma.Modern study proves, Ganoderma contains aminoacid, polypeptide, protein, and saccharide, ergosterol, triterpenes etc.Have and calm the nerves, regulate immunity, resist myocardial ischemia, anti-antiplatelet aggregation and antithrombotic, protect the liver, antioxidation, antiinflammatory, antitumor action.

What < < Chinese Pharmacopoeia > > recorded gathers and processing method, gather the whole year, remove impurity, wipe out the lower end stem with rotten wood, silt or culture matrix, dry in the shade or 40～50 ℃ of oven dry.

Summary of the invention

The drying means that the object of this invention is to provide a kind of Ganoderma, the Ganoderma that the method is dry, main effective ingredient polyoses content is high, and active anticancer is strong.

The present invention is achieved in that

The application of Refrigeration Technique in Ganoderma is dry, is characterized in that: get the Ganoderma of newly gathering, and freezing, the 2-5 days that banks up is dry.

The application of Refrigeration Technique of the present invention in Ganoderma is dry, is characterized in that: freezing is to be refrigerated to freeze.

The application of Refrigeration Technique of the present invention in Ganoderma is dry, is characterized in that: freezing is to be refrigerated to-5 ℃ to freeze below.

The application of Refrigeration Technique of the present invention in Ganoderma is dry, is characterized in that: dry is that heap postpone is dried or dries or dries.

Of the present inventionly being refrigerated to subzero certain ℃, is to be refrigerated to and to freeze at this subzero certain ℃.

So far completed the present invention, the Ganoderma of preparing by drying means of the present invention, main effective ingredient polyoses content is high, and active anticancer is strong.

Below by test example, further illustrate usefulness of the present invention, test example is intended to further illustrate usefulness of the present invention, but not limitation of the present invention.

By with a collection of Ganoderma of newly gathering, remove impurity, process by the following method.

1) dry: shady and cool ventilation place, spread out and dry;

2) dry: dry;

3) dry: 50 ℃ of following oven dry;

4) the freezing oven dry of banking up: be refrigerated to-5 ℃ to freezing, bank up 3 days at dark and damp place, 50 ℃ of following oven dry.

One, the dry ganoderma polyoses content comparison of distinct methods

The preparation of reference substance solution: get glucose reference substance appropriate, accurately weighed, add water and make every 1ml containing the solution of 0.1mg, obtain.

The preparation of standard curve: precision measures reference substance solution 0.2ml respectively, 0.4m1, 0.6ml, 0.8ml, 1.0ml, 1.2ml, put in 10ml tool plug test tube, add water to 2.0ml, precision adds sulphuric acid anthrone solution, and (precision takes anthrone 0.1g, adding 80% sulfuric acid solution 100ml makes to dissolve, shake up) 6ml, shake up, put in water-bath and heat 15 minutes, take out, put in people's ice bath cooling 15 minutes, take corresponding reagent as blank, according to < < Chinese Pharmacopoeia > > ultraviolet visible spectrophotometry, at 625nm wavelength place, measure absorbance, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.

The preparation of need testing solution: get the about 2g of each sample powder, accurately weighed, put in apparatus,Soxhlet's, add water 90ml, electric heater heating and refluxing extraction is to extracting liquid colourless, and extracting solution is transferred in 100ml measuring bottle, adds water to scale, shaking up. precision measures 10ml, adds ethanol 150ml, shakes up, place 12 hours for 4 ℃, take out, centrifugal, the supernatant that inclines, precipitation is dissolved in water, and is transferred in 50ml measuring bottle, add water to scale, shake up, obtain.

Algoscopy: precision measures need testing solution 2ml, put in 10ml tool plug test tube, method under the preparation of sighting target directrix curve, from " precision adds sulphuric acid anthrone solution 6ml ", measure absorbance in accordance with the law, from standard curve, read the weight (mg) that contains glucose in need testing solution, calculate, obtain.

Press dry product and calculate, contain ganoderan with anhydrous glucose (C ₆h1 ₂o ₆) meter, the results are shown in Table 1.

Table 1 distinct methods is dried ganoderma polyoses content (%)

Drying means	Dry	Dry	Dry	The freezing oven dry of banking up
					Content (%)	1.76	1.74	1.74	3.26

By table 1, found out, Ganoderma " freezing heap postpone is dry ", polyoses content is apparently higher than " drying ", " drying " and " oven dry ".

Two, the dry anti-human lung of Ganoderma source adenocarcinoma cell SPCA-1, human hepatoma cell line HepG2 and the people's chronic myelogenous leukemia cell K562 test of distinct methods.

1, prepare Ganoderma extract and get respectively the dry Ganoderma pulverizing of distinct methods, respectively decoct with water twice, add for the first time 8 times of amounts of water, decoct 2 hours, add for the second time 8 times of amounts of water, decoct 1 hour, merge medicinal liquid, medicinal liquid filters, and gets filtrate, and being concentrated into relative density is 1.10(60 ℃) time, add ethanol to make to reach 75% containing alcohol amount, precipitation, standing 36 hours, draw supernatant, reclaim ethanol, concentrated, dry, pulverize, obtain the dry Ganoderma extract of distinct methods.

2, tumor cell line people lung source adenocarcinoma cell SPCA-1, human hepatoma cell line HepG2 and people's chronic myelogenous leukemia cell K562, use RPMI-1649 culture fluid, and 37 ℃, 5%CO2, relative humidity 100% is cultivated, and attached cell 0.25% trypsinization goes down to posterity.

3, the inhibitory action of method MTT colorimetric method for determining extract to growth of tumour cell.The trophophase cell of taking the logarithm, is mixed with cell suspension with fresh RPMI-1640 culture fluid, 0.8 * 105/ml of 1 * 105/ml of suspension cell attached cell.Suspension cell is inoculated in 96 well culture plates after adding respectively variable concentrations Experimental agents; Attached cell is first inoculated in 96 well culture plates, after every hole 90 μ l24h, adds respectively variable concentrations Experimental agents.Final volume is that every hole 100 μ l and every group are established 3 parallel holes, establishes altogether 4 groups: be subject to reagent group T (culture fluid+cell suspension+variable concentrations is subject to reagent), negative control group N (culture fluid+cell suspension), medicine color matched group TC (culture fluid+variable concentrations is subject to reagent), blank group B (culture fluid+normal saline).Tested concentration is followed successively by 10 μ g/ml, 30 μ g/ml, 50 μ g/ml.Put 37 ℃, in 5%CO2 incubator, cultivate after 72h, after adding the 10 μ l concussions of MTT solution to mix to every hole, continue to cultivate 4h, add SDS90 μ l to stop cultivating, 37 ℃ are spent the night, and then under room temperature, on micro oscillator, shake 10min, microplate reader is measured the absorbance (OD value) at 570nm wavelength place, and experiment repeats 3 times.

Calculate as follows growth inhibition ratio:

Growth inhibition ratio (%)=(N group OD average-B group OD average)-(T group OD average-TC group OD average)/N group OD average-B group OD average * 100%.

4, different extracts to the suppression ratio of people's lung source adenocarcinoma cell SPCA-1, human hepatoma cell line HepG2 and people's chronic myelogenous leukemia cell K562 the results are shown in Table 2, table 3, table 4.

The suppression ratio result of table 2 couple SPCA-1

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	46.52	53.42	66.51
Dry	46.50	53.42	66.50
				Dry	46.47	53.50	66.53
The freezing oven dry of banking up	57.68	71.38	79.64

The suppression ratio result of table 3 couple HepG2

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	43.62	54.74	58.67
Dry	43.59	54.74	58.66
				Dry	43.61	54.71	58.64
The freezing oven dry of banking up	56.48	67.49	83.16

The suppression ratio result of table 4 couple K562

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	57.29	68.44	74.65
Dry	57.28	68.45	74.65
				Dry	57.31	68.42	74.62
The freezing oven dry of banking up	64.38	73.75	83.02

Result shows, the dry Ganoderma of distinct methods all has inhibitory action to the propagation of SPCA-1, HepG2, K562 cell, wherein, freezingly banks up that to dry curative effect best.

The specific embodiment

embodiment 1

Get the Ganoderma of newly gathering, put in freeze dryer, be refrigerated to-5 ℃ to freezing, at dark and damp place, bank up 2 days, dry.

embodiment 2

Get the Ganoderma of newly gathering, put in freeze dryer, be refrigerated to-10 ℃, bank up in the cool 3 days, dry.

embodiment 3

Get the Ganoderma of newly gathering, put in freeze dryer, be refrigerated to-20 ℃, indoor, bank up 4 days, dry.

embodiment 4

Get the Ganoderma of newly gathering, put in freeze dryer, be refrigerated to-32 ℃ to freezing, outdoor, bank up 3 days, dry.

embodiment 6

Get the Ganoderma of newly gathering, put in freeze dryer, be refrigerated to-15 ℃, at dark and damp place, bank up 5 days, dry.

Claims

1. the application of Refrigeration Technique in Ganoderma is dry, is characterized in that: get the Ganoderma of newly gathering, and freezing, the 2-5 days that banks up is dry.

2. the application in Ganoderma is dry according to the Refrigeration Technique of claim 1, is characterized in that: freezing is to be refrigerated to freeze.

3. the application in Ganoderma is dry according to the Refrigeration Technique of claim 1, is characterized in that: freezing is to be refrigerated to-5 ℃ to freeze below.

4. the application in Ganoderma is dry according to the Refrigeration Technique of claim 1, is characterized in that: dry is that heap postpone is dried or dries or dries.