CN104258106A - Preparation method of effective part of dendrobium officinale - Google Patents
Preparation method of effective part of dendrobium officinale Download PDFInfo
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Abstract
The invention discloses a preparation method of an effective part of dendrobium officinale. The method comprises steps as follows: firstly, extracting fat-soluble extracts represented by bibenzyl from the dendrobium officinale with a high-concentration alcohol-water system, refining the extracts with a macroporous resin method to obtain a total bibenzyl effective part with the bibenzyl content higher than 50%, and preparing dregs of the extracts into dendrobium officinale total polysaccharides with water extraction and alcohol precipitation methods; drying and evenly mixing the total bibenzyl effective part and the dendrobium officinale total polysaccharides obtained in two steps to obtain a dendrobium officinale total effective part extract. According to the method, the defect of the extremely low extraction rate of dendrobium officinale fat-soluble active ingredients represented by the bibenzyl with traditional methods such as a water decoction method, a tea making method and the like is overcome, the utilization efficiency of the effective part of the dendrobium officinale is improved substantially, pharmacological anti-oxidation animal experiments and anti-tumor cytotoxicity experiments prove that the activity of the effective part is significantly higher than that of traditional aqueous extracts, and the effective part has the characteristics of high extraction rate and high activity.
Description
Technical field
The present invention relates to medical art, specifically a kind of preparation method of Herba Dendrobii effective site.
Background technology
Herba Dendrobii is the orchid family Dendrobium herbaceos perennial, and its main medicinal part is fresh or dry stem, and being mainly distributed in the ground such as Anhui, Zhejiang, Guangxi, Hunan, Yunnan, Guizhou, is traditional rare Chinese medicine.Herba Dendrobii has reinforcing stomach reg fluid, effect of nourishing YIN and clearing away heat, loses, xerostomia excessive thirst, deficiency of stomach-Yin for cloudy impairment of body fluid, and lack of appetite is retched, and deficiency-heat is not moved back after being ill, hyperactivity of fire caused by deficiency of YIN, hectic fever due to YIN-deficiency consumptive fever, poor vision, and muscles and bones flaccidity is soft waits disease.Shennong's Herbal is classified as top grade, first of it is described as by Taoism regimen classics " Taoist Scriptures " " China's nine large Herba mesonae chinensis ".
Main containing water miscible polysaccharide, fat-soluble bibenzyl in Herba Dendrobii, also containing a small amount of water miscible aminoacid, water miscible trace element, water miscible phenolic acids, and the partially fat-soluble Herba Dendrobii alkaloid of trace, fat-soluble Herba Dendrobii sesquiterpene, partially fat-soluble Fluorenone compounds, partially fat-soluble lignanoid, water miscible phenols, partially fat-soluble flavone, fat-soluble steroidal etc., wherein bibenzyl has the valuable pharmacological such as antitumor, antioxidation active function.Traditional Herba Dendrobii Related product, comprise various little complex health care product, Herba Dendrobii extract of making tea, polysaccharide, its technique is water extraction or water extraction concentrated extract, the Bibenzyl compound and other liposoluble constituents that play the valuable pharmacological such as antitumor, antioxidation active function obviously can not well extract by this traditional water extraction process, cause that Product Process is extensive, weak curative effect, serious waste of resources.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of Herba Dendrobii effective site, substantially increase the extraction ratio of total polysaccharides and total bibenzyl.
For achieving the above object, the invention provides following technical scheme:
A preparation method for Herba Dendrobii effective site, comprises the extraction of fat-soluble total bibenzyl and the extraction of water solublity total polysaccharides.
As the further scheme of the present invention: the extraction step of described fat-soluble total bibenzyl is: high concentration ethanol aqueous solution Herba Dendrobii being added 8-12 times amount 65%-95% by solid-to-liquid ratio, is heated to 70-100 DEG C, reflux, extract, 2-3 time, each 2-3h; Merge extractive liquid, concentrating under reduced pressure obtains liposoluble extract; Dissolve this extract with ethanol in proper amount, then be added in macroporous resin column, get the ethanol water eluting part of its 25%-75%, concentrate and obtain total bibenzyl effective site; The extraction step of described water solublity total polysaccharides is: with the remaining medicinal residues of extraction step of fat-soluble total bibenzyl for raw material, add 8-12 times of water gaging, 70-100 DEG C of heating and refluxing extraction 2-3 time, each 2-3h, be concentrated into the volume of 1/10-1/12, the ethanol adding 4-6 times amount 95% carries out precipitate with ethanol, and filter, drying and crushing obtains total polysaccharides; By total bibenzyl effective site and total polysaccharides xeraphium Homogeneous phase mixing, obtain Herba Dendrobii total effective extractive part.
As the further scheme of the present invention: the extraction step of described fat-soluble total bibenzyl is: ethanol water Herba Dendrobii being added 12 times amount 80% by solid-to-liquid ratio, is heated to 92 DEG C, reflux, extract, 2 times, each 3h; Merge extractive liquid, concentrating under reduced pressure obtains liposoluble extract; Dissolve this extract with ethanol in proper amount, then be added in D101 macroporous resin column, get the ethanol water eluting part of its 40%-60%, concentrate and obtain total bibenzyl effective site.
Compared with prior art, the invention has the beneficial effects as follows: the mode such as overcome traditional water decoction, make tea is for the extremely low shortcoming of the Herba Dendrobii fat-soluble active ingredient extraction ratio taking bibenzyl as representative, significantly improve the utilization ratio of Herba Dendrobii effective ingredient, and through the zoopery of pharmacology's antioxidation and the experiment of antitumor cell poison, prove that its specific activity tradition water extract has to significantly improve, have high extraction, highly active feature concurrently.
Accompanying drawing explanation
Fig. 1 is the HPLC-DAD chromatogram of Herba Dendrobii liposoluble extract, and wavelength is 280nm;
Fig. 2 is the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract, and wavelength is 280nm;
Fig. 3 is the first mass spectrometric figure of chromatographic peak 1 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 4 is the first mass spectrometric figure of chromatographic peak 2 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 5 is the first mass spectrometric figure of chromatographic peak 3 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 6 is the first mass spectrometric figure of chromatographic peak 4 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 7 is the first mass spectrometric figure of chromatographic peak 5 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 8 is the first mass spectrometric figure of chromatographic peak 6 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Fig. 9 is the first mass spectrometric figure of chromatographic peak 7 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract;
Figure 10 is the first mass spectrometric figure of chromatographic peak 8 in the HPLC-MS liquid chromatogram of Herba Dendrobii liposoluble extract.
Detailed description of the invention
Below in conjunction with the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
In the embodiment of the present invention, a kind of preparation method of Herba Dendrobii effective site, comprises the extraction of fat-soluble total bibenzyl and the extraction of water solublity total polysaccharides.The extraction step of described fat-soluble total bibenzyl is: high concentration ethanol aqueous solution Herba Dendrobii being added 8-12 times amount 65%-95% by solid-to-liquid ratio, is heated to 70-100 DEG C, reflux, extract, 2-3 time, each 2-3h; Merge extractive liquid, concentrating under reduced pressure obtains liposoluble extract; Dissolve this extract with ethanol in proper amount, then be added in macroporous resin column, get the ethanol water eluting part of its 25%-75%, concentrate and obtain total bibenzyl effective site; The extraction step of described water solublity total polysaccharides is: with the remaining medicinal residues of extraction step of fat-soluble total bibenzyl for raw material, add 8-12 times of water gaging, 70-100 DEG C of heating and refluxing extraction 2-3 time, each 2-3h, be concentrated into the volume of 1/10-1/12, the ethanol adding 4-6 times amount 95% carries out precipitate with ethanol, and filter, drying and crushing obtains total polysaccharides; By total bibenzyl effective site and total polysaccharides xeraphium Homogeneous phase mixing, obtain Herba Dendrobii total effective extractive part.
Standards of pharmacopoeia method precipitate with ethanol-phenol-sulphuric acid colorimetry is adopted to measure total polysaccharides extraction ratio; Take EN as reference substance, adopt ultraviolet spectrophotometry under 280nm, measure total bibenzyl extraction ratio.
1, following two kinds of modes are adopted to detect the content of total bibenzyl in liposoluble extract
(1) HPLC-PAD method detects the Bibenzyl compound in liposoluble extract
Method: take out the centrifugal 10min of 4ml extracting solution 10000r, is redissolved with methanol after supernatant evaporate to dryness and transfers in auto injection bottle again, detected by HPLC-PAD.
Instrument: Waters 2695-2996Photodiode Array Detector high performance liquid chromatograph.
Chromatographic condition: COSMOSIL Packed ODS chromatographic column (250mm × 4.6mm); Column temperature 25 DEG C; Determined wavelength 280nm; Mobile phase is: methanol-0.1% formic acid (20:80); Flow velocity is 0.8ml/min.
Result: see accompanying drawing 1 and table 1.Chromatographic peak list in Fig. 1 and PAD UV scanning result, and the ultraviolet data of liposoluble extract and EN reference substance:
Conclusion: in conjunction with bibenzyl UV characteristic absorption peak at about 280nm, 220nm, illustrates in this liposoluble extract containing a large amount of compounds with bibenzyl UV characteristic absorption peak.
Table 1PAD UV scanning result
| Retention time | Peak area | Peak area percent | Ultraviolet maximum absorption band/nm | |
| 1 | 2.519 | 59033 | 0.34 | 294 |
| 2 | 2.863 | 1443223 | 8.37 | 288 |
| 3 | 3.164 | 372468 | 2.16 | 278(shoulder?peak) |
| 4 | 3.518 | 1688226 | 9.79 | 271 |
| 5 | 3.862 | 883337 | 5.12 | 257 |
| 6 | 4.067 | 129686 | 0.75 | 275(shoulder?peak) |
| 7 | 4.249 | 2179418 | 12.64 | 280 |
| 8 | 4.699 | 952448 | 5.52 | 295 |
| 9 | 5.247 | 4531132 | 26.28 | 287 |
| 10 | 5.636 | 2886638 | 16.74 | 284 |
| 11 | 7.143 | 157225 | 0.91 | 300 |
| 12 | 7.825 | 87928 | 0.51 | 276,220 |
| 13 | 8.068 | 54018 | 0.31 | 280,218 |
| 14 | 8.716 | 190555 | 1.11 | 285 |
| 15 | 9.474 | 742223 | 4.31 | 276 |
| 16 | 11.72 | 78187 | 0.45 | 293 |
| 17 | 12.622 | 106621 | 0.62 | 300 |
| 18 | 13.871 | 78581 | 0.46 | 259,206 |
| 19 | 14.967 | 24184 | 0.14 | 283,253 |
| 20 | 15.333 | 109394 | 0.63 | 305,210 |
| 21 | 17.1 | 44359 | 0.26 | 285,223 |
Continued 1PAD UV scanning result
| Retention time | Peak area | Peak area percent | Ultraviolet maximum absorption band/nm | |
| 22 | 17.483 | 82995 | 0.48 | 293,234 |
| 23 | 18.755 | 165076 | 0.96 | 291,248 |
| 24 | 21.6 | 80513 | 0.47 | 281,228 |
| 25 | 23.357 | 112784 | 0.65 | 283,222 |
| Total position | 278,220 | |||
| EN | 280,218 |
(2) HPLC-MS method detects the Bibenzyl compound in liposoluble extract
Method: take out the centrifugal 10min of 4ml extracting solution 10000r, is redissolved with methanol after supernatant evaporate to dryness and transfers in auto injection bottle again, detected by HPLC-MS.
Instrument: Aglient 1290 liquid chromatograph-G6200 series of time of flight GC-MS.
Chromatographic condition: methanol-water (30:70); Flow velocity is 0.8ml/min.
Result: as shown in figs. 2-10.
Conclusion: in HPLC-MS liquid chromatogram, there is the characteristic molecular amount of bibenzyl series in the first mass spectrometric figure of main chromatographic peak, if mass-to-charge ratio m/z is 304 (Fig. 3), 288 (Fig. 4), 342 (Fig. 5), 198 (Fig. 6), 278 (Fig. 7-Fig. 8), 378 (Fig. 9), 420 (Figure 10) etc.Containing a large amount of Bibenzyl compound in this extract visible.
2, the optimization of orthogonal test of liposoluble extract extraction conditions
Select to carry out positive quadraturing design test to the larger concentration of alcohol of bibenzyl extraction ratio impact, extraction time and extraction time three factors, with bibenzyl extraction ratio for index, preferably best technological parameter, determines to take ethanol as the optimum extraction process of solvent extraction bibenzyl.Factor level arranges as table 2.
Concrete steps are: take Herba Dendrobii 1g, carry out circumfluence method extraction, and fixing heating-up temperature is 92 DEG C, and concentration of alcohol is 80%, and ultraviolet spectrophotometry method measures total bibenzyl content.Orthogonal experiments and analysis in table 3-table 4.
Orthogonal experiment results shows, the influence degree of each factor is followed successively by B>C>A, i.e. extraction time > extraction time > concentration of alcohol, and optimum extraction process is B
3c
3a
3, i.e. the ethanol water of 80%, reflux, extract, 3 times, each 3h.
Table 2 orthogonal test factor level table
The bibenzyl extraction ratio result of table 3 orthogonal test
The variance analysis of table 4 orthogonal test
Demonstration test:
Take Herba Dendrobii 1g, add the ethanol water of 12ml80%, soaked overnight, 92 DEG C of heating and refluxing extraction 2 times, let cool, are transferred in 25ml volumetric flask, with a small amount of moisture time washing container, washing liquid is incorporated in same volumetric flask, adds water to scale, shake up, get the centrifugal 15min of 4ml wherein, rotating speed is 10000r/min, precision measures supernatant and dilutes 20 times, record A at 280nm place, then read C from regression equation, calculate bibenzyl extraction ratio.Result is as table 5: the optimum extraction rate of Bibenzyl compound is 3.25%, does not have on the basis of significant difference, in order to energy savings at extraction ratio, the second extracting method can be selected, namely 12 times amount 80% ethanol water 92 DEG C extraction, extract 2 times, each 3h, extraction rate reached is to 3.11%.
Bibenzyl extraction ratio in table 5 demonstration test
3, matched group one: the extraction ratio of total polysaccharides and total bibenzyl in the method that tradition brews up
1) repeatedly to make tea experiment: take Herba Dendrobii 6g, add boiling water 300ml, soak once every 2h, last soaked overnight.Result is as shown in table 6, and it is 1.50% that total polysaccharides adds with total extraction ratio, and it is 1.72% that total bibenzyl adds with total extraction ratio.
2) single long-time heat preservation bottle/thermos-cooker immersion test: take Herba Dendrobii 3g, put into thermos flask/thermos-cooker, add boiling water 150ml, sealed thermal insulating soaks 10h, and the extraction ratio of total polysaccharides is 1.19%, and the extraction ratio of total bibenzyl is 1.37%.
Table 6 is made tea for many times the total polysaccharides of experiment and the extraction ratio of total bibenzyl
4, matched group two: the extraction ratio of total bibenzyl in Herba Dendrobii in traditional extraction process by water.
The total polysaccharides of the traditional extraction process by water of table 7 and the extraction ratio of total bibenzyl
Take Herba Dendrobii 1g, 100 DEG C, the water adding 12 times amount extracts 2 times, each 2h.Result is as shown in table 7: the average recoveries of total polysaccharides is 5.98%, and the average recoveries of total bibenzyl is 1.74%.
In sum: contrast three kinds of different extracting modes, traditional makes tea in method, repeatedly brew adduction extraction ratio and once to brew extraction ratio for a long time suitable, but the latter taking dose is high due to one time, possible drug effect is better; Tradition extraction process by water is with to brew mode the same, and the extraction ratio for liposoluble constituent is relative not high; The present invention, by the alcohol extraction of high concentration, significantly improves the extraction ratio of Bibenzyl compound.
Embodiment 1
The preparation of total bibenzyl effective site
Dry Herba Dendrobii 1kg, adds the ethanol of 12L80%, soaked overnight, 92 DEG C of heating and refluxing extraction 2 times, each 3h, merge extractive liquid, and recycling design, to obtaining 98g extractum without alcohol taste and drying, is liposoluble extract; With the dissolve with ethanol of 95% of 30ml, then be put on D101 type macroporous resin, get 40%-60% eluting position, concentrating under reduced pressure is dry, and obtain total bibenzyl effective site 46g, the ultraviolet spectrophotometry bibenzyl content recorded in effective site is 60.5%.
Embodiment 2
The preparation of Herba Dendrobii total effective parts
Example 1 Chinese medicine slag prepares total polysaccharides by decoction and alcohol sedimentation technique, and namely the water of 12L is heated to 100 DEG C of extractions 2 times, each 2h; Extracting solution is concentrated into 1/12 volume 2L, adds the ethanol of 10L 95%, precipitate polysaccharides, filter, dry, obtain total polysaccharides 42g, get above-mentioned total bibenzyl 23g, merge with total polysaccharides 21g, obtain total effective parts 44g, be equivalent to the amount of crude drug 500g.
Embodiment 3
Tradition water extract, total effective parts, the activity research of total bibenzyl effective site
3.1 antioxidation zooperies
Animal: Kunming mouse, is provided by Zhejiang University's Experimental Animal Center.18 months ages of Mus, body weight 45 ~ 55g, totally 40, male and female half and half.
Male and female mice is divided into 4 groups respectively at random: normal aged mouse matched group; Tradition water extract group; Total effective parts group; Total bibenzyl effective site group.
Every day gavage 2 times, each 50mg, continuous 30d.2h after last administration, sacrificed by decapitation mice, gets blood, separated plasma and erythrocyte; Separately get hepatic tissue, nephridial tissue and ventricular homogenate, measure Plasma MDA (MDA) by sulfo-barbital method, measure the activity of erythrocyte superoxide dismutase (SOD) by EistnerFF method, experimental result is as shown in table 8.
Table 8 Herba Dendrobii extract is on the impact (* P<0.01) of Antioxidation Effect
Conclusion: compare traditional water extract, total effective parts and total bibenzyl effective site obviously can increase the drug effect of antioxidation.
3.2 antitumor cell poison experiments
Observe candidate compound to the inhibitory action of the in-vitro multiplication ability of human lung carcinoma cell 95-D and Proliferation of Human Ovarian Cell A2780.
Method: Sulforhodamine B (SRB) method detects candidate compound to the inhibitory action of 95-D and A2780 multiplication capacity being in exponential phase.
Experiment material: tested extract, tetrazolium bromide SRB (Sigma), experiment human lung carcinoma cell line 95-D and Proliferation of Human Ovarian Cell A2780.95-D and A2780 is incubated at RPMI-1640 and DMEM (GIBCO respectively, Grand Island, NY, U.S.A) containing the hyclone of 10% deactivation, L-glutaminate (GIBCO) 2nmol/L, penicillin 100IU/mL and streptomycin 100 μ g/mL.
Concrete steps: (1) adopts cell strain: human lung carcinoma cell 95-D and Proliferation of Human Ovarian Cell A2780; (2) by after the cell dissociation of exponential phase, blow and beat into single cell suspension, be inoculated in 96 well culture plates, 5x10
3cells/well, every hole culture medium 100 μ L, at 37 DEG C, the CO of 5%
2overnight incubation in incubator; (3) after cell attachment, the tested extract adding debita spissitudo cultivates 3 days again in incubator; (4) trichloroacetic acid adding 10% fixes 1h; (5) with distilled water washing, after drying, every hole adds the SRB solution of 100 μ L4mg/mL, and room temperature dyes 20 minutes, with the acetic acid washing of 1%, dry; (6) Tris (Tris) solution that every hole adds the 10mM of 100 μ L makes SRB dissolve; (7) microplate reader detects each hole OD value, and determined wavelength is 515nm; Record result; By following formulae discovery suppression ratio: suppression ratio (%)=(OD contrasts-OD administration)/OD contrasts × 100%, and calculates IC
50.If the IC of tested material
50littlely show that activity is better.
Experimental result is as shown in table 9:
Table 9 Herba Dendrobii extract Cyto toxic experiment showed (IC
50, μ g/ml)
Interpretation of result: tested extract breeds the inhibitory action had in various degree to human lung carcinoma cell 95-D and Proliferation of Human Ovarian Cell A2780, wherein total effective parts and total more traditional water extract drug effect of bibenzyl effective site significantly improve.
Embodiment 4
The preparation of total effective parts, total bibenzyl effective parts formulation
4.1 total effective parts make tablet
Mixed by embodiment 1 gained total effective parts 25g and starch 15g, add 10% starch slurry 5g and make soft material, add magnesium stearate 0.25g, dried starch 4g is pressed into 120 after mixing, and to obtain final product.Every sheet is containing total effective parts 208mg.
4.2 total effective parts make capsule
Embodiment 1 gained whole part extract 10g is added 10% starch slurry and makes soft material, after granulating with 14 mesh sieves, to put after 60 DEG C ~ 70 DEG C dryings in 12 mesh sieve granulate, be inserted in No. 1 Capsules.Fill grain 500 altogether, every capsules is containing total effective parts 20mg.
4.3 total bibenzyl effective sites make capsule
By total for embodiment 2 gained bibenzyl effective site 10g, at 60 DEG C ~ 70 DEG C drying rear mistake 12 mesh sieves, be inserted in No. 1 Capsules.Fill grain 200 altogether, every capsules is containing total bibenzyl effective site 50mg.
The mode such as instant invention overcomes traditional water decoction, make tea is for the extremely low shortcoming of the Herba Dendrobii fat-soluble active ingredient extraction ratio taking bibenzyl as representative, significantly improve the utilization ratio of Herba Dendrobii effective ingredient, and through the zoopery of pharmacology's antioxidation and the experiment of antitumor cell poison, prove that its specific activity tradition water extract has to significantly improve, have high extraction, highly active feature concurrently.
To those skilled in the art, obviously the invention is not restricted to the details of above-mentioned one exemplary embodiment, and when not deviating from spirit of the present invention or basic feature, the present invention can be realized in other specific forms.Therefore, no matter from which point, all should embodiment be regarded as exemplary, and be nonrestrictive, scope of the present invention is limited by claims instead of above-mentioned explanation, and all changes be therefore intended in the implication of the equivalency by dropping on claim and scope are included in the present invention.
In addition, be to be understood that, although this description is described according to embodiment, but not each embodiment only comprises an independently technical scheme, this narrating mode of description is only for clarity sake, those skilled in the art should by description integrally, and the technical scheme in each embodiment also through appropriately combined, can form other embodiments that it will be appreciated by those skilled in the art that.
Claims (3)
1. a preparation method for Herba Dendrobii effective site, is characterized in that, comprises the extraction of fat-soluble total bibenzyl and the extraction of water solublity total polysaccharides.
2. the preparation method of Herba Dendrobii effective site according to claim 1, it is characterized in that, the extraction step of described fat-soluble total bibenzyl is: high concentration ethanol aqueous solution Herba Dendrobii being added 8-12 times amount 65%-95% by solid-to-liquid ratio, be heated to 70-100 DEG C, reflux, extract, 2-3 time, each 2-3h; Merge extractive liquid, concentrating under reduced pressure obtains liposoluble extract; Dissolve this extract with ethanol in proper amount, then be added in macroporous resin column, get the ethanol water eluting part of its 25%-75%, concentrate and obtain total bibenzyl effective site; The extraction step of described water solublity total polysaccharides is: with the remaining medicinal residues of extraction step of fat-soluble total bibenzyl for raw material, add 8-12 times of water gaging, 70-100 DEG C of heating and refluxing extraction 2-3 time, each 2-3h, be concentrated into the volume of 1/10-1/12, the ethanol adding 4-6 times amount 95% carries out precipitate with ethanol, and filter, drying and crushing obtains total polysaccharides; By total bibenzyl effective site and total polysaccharides xeraphium Homogeneous phase mixing, obtain Herba Dendrobii total effective extractive part.
3. the preparation method of Herba Dendrobii effective site according to claim 2, it is characterized in that, the extraction step of described fat-soluble total bibenzyl is: ethanol water Herba Dendrobii being added 12 times amount 80% by solid-to-liquid ratio, is heated to 92 DEG C, reflux, extract, 2 times, each 3h; Merge extractive liquid, concentrating under reduced pressure obtains liposoluble extract; Dissolve this extract with ethanol in proper amount, then be added in D101 macroporous resin column, get the ethanol water eluting part of its 40%-60%, concentrate and obtain total bibenzyl effective site.
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