CN103948647A - Application of freezing technology in drying inonotus obliquus - Google Patents

Abstract

The invention relates to application a freezing technology in drying inonotus obliquus. The inonotus obliquus is prepared by selecting newly harvested inonotus obliquus, freezing, piling for 2-5 days and drying. According to the inonotus obliquus, the main effective components are high in polysaccharide content and strong in anti-cancer activity.

Description

The application of Refrigeration Technique in Inonqqus obliquus is dry

Invention field

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

Background technology

Traditional Chinese medicine drying has very long history in China, the Chinese herbal medicine of drying can keep certain medicinal ingredient and be difficult for putrid and deteriorated, the dry important measures as ensureing Chinese herbal medicine quality, it is a requisite technical process in Chinese herbal medicine processing, it is in close relations that dry and Chinese medicine are produced, and dry quality will directly affect the quality of product.

Inonqqus obliquus belongs to Eumycota, Basidiomycotina, Hymenomycetes, non-brown Zoopagales, Polyporaceae, brown transverse hole fungus genus, formal name used at school: Fuscoporia obliqua (Pers.Fr.) Aosh or Inonotus obliquus (Fr.) Pilat.From 16th century so far, the bacterium of always applying this bacterium among the people of some countries of Eastern Europe comes anti-curing cancers, heart disease, hepatopathy, gastropathy etc.In recent years, the research of this bacterium is caused to the relevant scholar's of a lot of countries attention, isolate the compositions such as the acid of bolt bacterium, triterpenoid compound, steroid, polysaccharide, and for prevention and the treatment of malignant tumor, diabetes, cardiovascular disease, hepatopathy and acquired immune deficiency syndrome (AIDS).

What " Shandong Province's Chinese crude drug standard " recorded gathers and processing method, for gathering the whole year, dries in the shade or 40-50 DEG C of oven dry.

Summary of the invention

The object of this invention is to provide the dry new method of a kind of Inonqqus obliquus, the Inonqqus obliquus 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 Inonqqus obliquus is dry, is characterized in that: get the Inonqqus obliquus of newly gathering, and freezing, the 2-5 days that banks up is dry.

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

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

The application of Refrigeration Technique of the present invention in Inonqqus obliquus 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 DEG C, is to be refrigerated to and to freeze at this subzero certain DEG C.

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

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

By with a collection of Inonqqus obliquus 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 DEG C of following oven dry;

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

One, the dry Fuscoporia obliqua polysaccharide content comparison of distinct methods

Take respectively each sample powder (60 order) 5g, add 150mL distilled water mix homogeneously, reflux, extract, 3h, cooling, the centrifugal 15min of 6000r/min, get supernatant, precipitation part repeats to extract twice again, merges supernatant, rotation vacuum concentration is to 1/10 of original volume, add ethanol to concentration of alcohol 80%, hold over night, collecting precipitation; Absolute ethanol washing, vacuum drying, obtains crude polysaccharides.

Get a certain amount of dried crude polysaccharides, be settled to certain volume with distilled water, adopt phenol-sulfuric acid process to measure polyoses content, with glucose as a standard product drawing standard curve, in dry product, the results are shown in Table 1.

Table 1 distinct methods is dried Fuscoporia obliqua polysaccharide content (%)

Drying means	Dry	Dry	Dry	The freezing oven dry of banking up
					Content (%)	5.31	5.24	5.26	9.37

Found out by table 1, Inonqqus obliquus " freezing heap postpone is dry ", polyoses content is apparently higher than " drying ", " drying " and " oven dry ".

Two, the dry anti-human lung of Inonqqus obliquus 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 Inonqqus obliquus extract and get respectively the dry Inonqqus obliquus pulverizing of distinct methods, respectively decoct with water twice, add water for the first time 8 times and measure, decoct 2 hours, add water for the second time 8 times and measure, decoct 1 hour, merge medicinal liquid, medicinal liquid filters, and gets filtrate, and being concentrated into relative density is 1.10(60 DEG C) time, add ethanol to make to reach 75% containing alcohol amount, precipitation, leaves standstill 36 hours, draw supernatant, reclaim ethanol, concentrated, dry, pulverize, obtain the dry Inonqqus obliquus 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 DEG C, 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, 0.8 × 105/ml of 1 × 105/ml of suspension cell attached cell with fresh RPMI-1640 culture fluid.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 DEG C, 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 DEG C 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 to SPCA-1

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	53.21	61.15	75.33
Dry	53.34	61.21	75.30
				Dry	53.21	61.17	75.34
The freezing oven dry of banking up	64.36	75.32	84.86

The suppression ratio result of table 3 to HepG2

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	43.85	54.20	60.27
Dry	43.82	54.21	60.27
				Dry	43.84	54.21	60.25
The freezing oven dry of banking up	55.74	63.48	86.51

The suppression ratio result of table 4 to K562

Suppression ratio (%)	10（μg/ml）	30（μg/ml）	50（μg/ml）
				Dry	55.52	70.44	75.78
Dry	55.51	70.43	75.86
				Dry	55.53	70.43	75.85
The freezing oven dry of banking up	71.43	76.82	81.27

Result shows, the dry Inonqqus obliquus 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.

Detailed description of the invention

embodiment 1

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

embodiment 2

Get the Inonqqus obliquus of newly gathering, put in freeze dryer, be refrigerated to-10 DEG C, bank up 3 days at dark and damp place, dry.

embodiment 3

Get the Inonqqus obliquus of newly gathering, put in freeze dryer, be refrigerated to-20 DEG C, bank up in the cool 4 days, dry.

embodiment 4

Get the Inonqqus obliquus of newly gathering, put in freeze dryer, be refrigerated to-32 DEG C to freezing, bank up 3 days indoor, dry.

embodiment 6

Get the Inonqqus obliquus of newly gathering, put in freeze dryer, be refrigerated to-15 DEG C, bank up 5 days outdoor, dry.

Claims (4)

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

2. the application in Inonqqus obliquus 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 Inonqqus obliquus is dry according to the Refrigeration Technique of claim 1, is characterized in that: freezing is to be refrigerated to-5 DEG C to freeze below.

4. the application in Inonqqus obliquus 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.