CN1951391A - Method for preparing suspension of rhodiola root nanoliposome - Google Patents

Abstract

The invention relates to a method for preparing gadol glycosides nanometer liposome suspension. Wherein, said suspension comprises the extractive of gadol as gadol glycosides and the nanometer liposome packing the gadol glycosides. The inventive production comprises that: dissolving the lecithin and cholesterol into absolute ether, depressurizing and vaporizing the organic solvent; adding gadol glycosides phosphate buffer with some surface activator, expanding, and dispersing at ultrasonic wave, to obtain the final suspension. The average grainness of product is lower than 100nm; the package rate of gadol glycosides can reach 25-35%.

Description

A kind of preparation method of suspension of rhodiola root nanoliposome

Technical field

A kind of preparation method of suspension of rhodiola root nanoliposome, function of dominant Sexual health promotion food technology field.

Background technology

Radix Rhodiolae (Rhodiola sachalinensis A.Bor.) is the east traditional herbal medicine that just is much accounted of nearest decades, and it is another the important health care medicine source plant resource that is developed after Radix Ginseng, Radix Et Caulis Acanthopanacis Senticosi.Main effective ingredient is rhodioside (Salidroside) and aglycon thereof, promptly to butyl alcohol (ρ-Tyrosol), human body had dual regulation, and without any side effects or one-tenth is practised addiction, has the metabolism of the human body energy of promotion, biological effects such as excitement, endocrine regulation function and antitoxin detoxifcation excite nerve.Existing in recent years many research report proof rhodiosides have antifatigue effect, anti-hypoxia, resisting microwave radiation effect, suppress health-care effecies such as blood sugar increasing, anti-peroxidation effect.The health product of Radix Rhodiolae class are extensively taken by the practitioner of special industries such as spaceman, mountaineer, driver.In the three class article lists that Ministry of Public Health in 2002 is determined, Radix Rhodiolae is listed in one of article list that can be used as health food.As seen, Radix Rhodiolae has exploitation potential as the research of Chinese medicine and health food.But there are many problems in commercially available rhodioside health product, fluctuated such as concentration in vivo, do not have the slow release effect, take that number of times is frequent, bioavailability is low or the like.

Up to now, do not see the report for preparing relevant for rhodiola root nanoliposome both at home and abroad.Though many researchers have begun to attempt rhodioside is carried out the phosphatide complexes of sodium alginate-chitosan microcapsules embedding and preparation pharmaceutical injection.Domestic scholars Zhao Wu is strange, and to adopt nontoxic, good biocompatibility, biodegradable natural macromolecular material sodium alginate and chitosan be the film material, prepared the slow releasing capsule of rhodioside.But the purity of the used rhodioside of this kind method one side is low, only the crude extract with rhodioside carries out embedding, contains a large amount of starch, albumen, tannin, natural gum, polarity pigment or the like impurity in the crude extract, on the other hand, the prepared microcapsule particle diameter is very big, and slow-release function is still not obvious.Exploitation as new medicinal preparation, people such as Gu Yanli attempt purified rhodioside is prepared the phosphatide complexes of rhodioside, the result of study of pharmacokinetics shows, the distribution characteristics of Radix Rhodiolae phosphatide complexes is compared with Radix Rhodiolae extract bigger variation has been taken place, and its drug plasma initial concentration obviously reduces.Further do the also proof that distributes in the mice body, the rhodioside of injection Isodose, the two rhodioside concentration in the mice internal organs is different, especially the rhodioside concentration that distributes in the heart of mice, brain, injection Radix Rhodiolae extract phosphatide complexes Emulsion group is obviously than injection Radix Rhodiolae extract component cloth concentration height.But this phosphatide complexes is only as the medicine injection, and used a large amount of poloxamers and the medical emulsifying agent of injection wet goods in the preparation, and safety hidden danger is bigger, is not suitable in the food industry.

Summary of the invention

The preparation method that the purpose of this invention is to provide a kind of rhodiola root nanoliposome, for main component-rhodioside of strengthening Radix Rhodiolae bioavailability as healthy ingredient, reach slow release or controlled-release effect, enhancing targeting in vivo is to improve health-care effecies such as its resisting fatigue, radioprotective.

Technical scheme of the present invention: the preparation method of suspension of rhodiola root nanoliposome is the nanometer liposome that comprises rhodiola active ingredient extract rhodioside in this suspension and seal rhodioside.With the rhodioside extracting solution through macroporous adsorbent resin, hyperfiltration treatment is core, and lecithin, cholesterol are the film material, are aided with Tween 80, adopts ultrasonic method to prepare rhodiola root nanoliposome.

(1) preparation rhodioside phosphate buffer: the Radix Rhodiolae powder with mistake 60 mesh sieves is a raw material, make to extract solvent with 20% (v/v) ethanol, solid-to-liquid ratio with g/mL count 1: 25～30, preimpregnation 60 minutes, adopt microwave auxiliary extraction technology, microwave power is 130W, microwave irradiation time 1 minute, repetitive operation is extracted once, merge the crude extract of secondary, the crude extract of gained rhodioside is again through the DA201-C absorption with macroporous adsorbent resin, with 50% (v/v) ethanol elution of 4 times of column volumes, thin up behind the concentrated recovery of the eluent of the collecting ethanol, control rhodioside concentration is 1mg/mL, through the ultrafilter membrane ultrafiltration of molecular cut off 3000Da, is concentrated into the filtered solution of collecting dried, use pH7.0,0.05mol/L phosphate buffer preparation rhodioside solution, the concentration of control rhodioside solution is 0.5mg/mL.

(2) lipid film forming: lecithin, cholesterol are dissolved in a small amount of absolute ether by mass ratio at 5: 1, be transferred in the rotary evaporation bottle, 40 ℃ of rotary evaporation organic solvents form uniform one deck lipid membrane on the bottle wall, and keep decompression 30 minutes, to remove remaining ether;

(3) hydration swelling: Tween 80 is added in the rhodioside phosphate buffer of preparation, the concentration that makes Tween 80 in the buffer is 1% (w/w), the lecithin quality is counted 1: 100 than buffer volume with g/mL, gets this solution and rotates hydration 30 minutes down at 40 ℃;

(4) ultra-sonic dispersion: the suspension after the hydration is handled through probe type ultrasonic again, and ultrasound intensity 50%～70% was opened in 1 second, and 1 second stops, and handles 5～7 minutes, promptly gets the suspension of rhodiola root nanoliposome.

The mensuration of particle diameter: adopt the dynamic light scattering particle size instrument to measure, measure 25 ℃ of temperature, diluted sample is to phospholipid concentration 0.025%.The rhodioside envelop rate adopts dialysis to measure.

The mean diameter of the rhodiola root nanoliposome that makes is less than 100nm, and the envelop rate of rhodioside can reach 25%～35%.

Beneficial effect of the present invention: with the rhodioside extracting solution through macroporous adsorbent resin, hyperfiltration treatment is core, and lecithin, cholesterol are the film material, are aided with Tween 80, adopts ultrasonic method to prepare rhodiola root nanoliposome.Can strengthen the bioavailability of the main component-rhodioside of Radix Rhodiolae as healthy ingredient, reach slow release or controlled-release effect, enhancing targeting in vivo is to improve health-care effecies such as its resisting fatigue, radioprotective.

Description of drawings

The particle size distribution figure of Fig. 1 rhodiola root nanoliposome

The specific embodiment

Embodiment 1

Accurately take by weighing 60 order Radix Rhodiolae powder 50g,, under microwave power 130W, microwave irradiation time 60s, extract 1min with 1450mL20% (v/v) ethanol preimpregnation 60min, filter crude extract.Filtering residue still extracts once by above operation, merges the crude extract of secondary, concentrates to reclaim ethanol.The crude extract of gained rhodioside is again through the DA201-C absorption with macroporous adsorbent resin, 50% (v/v) ethanol elution of 4 times of column volumes, thin up to rhodioside concentration is 1mg/mL behind the concentrated recovery of the eluent of the collecting ethanol, the ultrafiltration membrane treatment of reuse molecular cut off 3000Da, the filtered solution of collecting is concentrated into dried, is made into phosphate buffer (pH7.0).Accurately take by weighing lecithin 200mg, cholesterol 40mg, be dissolved in the 5mL absolute ether, be transferred in the 250mL rotary evaporation bottle, 40 ℃ of rotary evaporations are removed ether, form uniform one deck lipid membrane on the bottle wall, and keep decompression 30min, to remove remaining ether.Add rhodioside phosphate buffer (pH7.0) 20mL that contains 1% (w/w) Tween 80 in evaporative flask, 40 ℃ are rotated hydration 30min down.The hydration volume is 100 times of lecithin quality, and rhodioside concentration is 0.5mg/mL.Suspension after the hydration is handled (intensity 70%, 1s opens, 1s stops) through the probe type ultrasonic of 5min again, promptly gets the suspension of rhodiola root nanoliposome.

Embodiment 2

Accurately take by weighing 60 order Radix Rhodiolae powder 50g,, under microwave power 130W, microwave irradiation time 60s, extract 1min with 1250mL20% (v/v) ethanol preimpregnation 60min, filter crude extract.Filtering residue still extracts once by above operation, merges the crude extract of secondary, concentrates to reclaim ethanol.The crude extract of gained rhodioside is again through the DA201-C absorption with macroporous adsorbent resin, 50% (v/v) ethanol elution of 4 times of column volumes, thin up to rhodioside concentration is 1mg/mL behind the concentrated recovery of the eluent of the collecting ethanol, the ultrafiltration membrane treatment of reuse molecular cut off 3000Da, the filtered solution of collecting is concentrated into dried, is made into phosphate buffer (pH7.0).Accurately take by weighing lecithin 100mg, cholesterol 20mg, be dissolved in the 5mL absolute ether, be transferred in the 250mL rotary evaporation bottle, 40 ℃ of rotary evaporations are removed ether, form uniform one deck lipid membrane on the bottle wall, and keep decompression 30min, to remove remaining ether.Add rhodioside phosphate buffer (pH7.0) 10mL that contains 1% (w/w) Tween 80 in evaporative flask, 40 ℃ are rotated hydration 30min down.The hydration volume is 100 times of lecithin quality, and rhodioside concentration is 0.5mg/mL.Suspension after the hydration is handled (intensity 70%, 1s opens, 1s stops) through the probe type ultrasonic of 5min again, promptly gets the suspension of rhodiola root nanoliposome.

Embodiment 3

Accurately take by weighing 60 order Radix Rhodiolae powder 50g,, under microwave power 130W, microwave irradiation time 60s, extract 1min with 1500mL20% (v/v) ethanol preimpregnation 60min, filter crude extract.Filtering residue still extracts once by above operation, merges the crude extract of secondary, concentrates to reclaim ethanol.The crude extract of gained rhodioside is again through the DA201-C absorption with macroporous adsorbent resin, 50% (v/v) ethanol elution of 4 times of column volumes, thin up to rhodioside concentration is 1mg/mL behind the concentrated recovery of the eluent of the collecting ethanol, the ultrafiltration membrane treatment of reuse molecular cut off 3000Da, the filtered solution of collecting is concentrated into dried, is made into phosphate buffer (pH7.0).Accurately take by weighing lecithin 200mg, cholesterol 40mg, be dissolved in the 5mL absolute ether, be transferred in the 250mL rotary evaporation bottle, 40 ℃ of rotary evaporations are removed ether, form uniform one deck lipid membrane on the bottle wall, and keep decompression 30min, to remove remaining ether.Add rhodioside phosphate buffer (pH7.0) 20mL that contains 1% (w/w) Tween 80 in evaporative flask, 40 ℃ are rotated hydration 30min down.The hydration volume is 100 times of lecithin quality, and rhodioside concentration is 0.5mg/mL.Suspension after the hydration is handled (intensity 50%, 1s opens, 1s stops) through the probe type ultrasonic of 7min again, promptly gets the suspension of rhodiola root nanoliposome.

Claims (1)

1, a kind of preparation method of suspension of rhodiola root nanoliposome, the nanometer liposome that it is characterized in that comprising rhodiola active ingredient extract rhodioside in this suspension and seal rhodioside, preparation technology is:

(1) preparation rhodioside phosphate buffer: the Radix Rhodiolae powder with mistake 60 mesh sieves is a raw material, make to extract solvent with 20% (v/v) ethanol, solid-to-liquid ratio with g/mL count 1: 25～30, preimpregnation 60 minutes, adopt microwave auxiliary extraction technology, microwave power is 130W, microwave irradiation time 1 minute, repetitive operation is extracted once, merge the crude extract of secondary, the crude extract of gained rhodioside is again through the DA201-C absorption with macroporous adsorbent resin, with 50% (v/v) ethanol elution of 4 times of column volumes, thin up behind the concentrated recovery of the eluent of the collecting ethanol, control rhodioside concentration is 1mg/mL, through the ultrafilter membrane ultrafiltration of molecular cut off 3000Da, is concentrated into the filtered solution of collecting dried, use pH7.0,0.05mol/L phosphate buffer preparation rhodioside solution, the concentration of control rhodioside solution is 0.5mg/mL;

(2) lipid film forming: lecithin, cholesterol are dissolved in a small amount of absolute ether by mass ratio at 5: 1, be transferred in the rotary evaporation bottle, 40 ℃ of rotary evaporation organic solvents form uniform one deck lipid membrane on the bottle wall, and keep decompression 30 minutes, to remove remaining ether;

(3) hydration swelling: Tween 80 is added in the rhodioside phosphate buffer of preparation, the concentration that makes Tween 80 in the buffer is 1% (w/w), the lecithin quality is counted 1: 100 than buffer volume with g/mL, gets this solution and rotates hydration 30 minutes down at 40 ℃;

(4) ultra-sonic dispersion: the suspension after the hydration is handled through probe type ultrasonic again, and ultrasound intensity 50%～70% was opened in 1 second, and 1 second stops, and handles 5～7 minutes, promptly gets the suspension of rhodiola root nanoliposome.