CN101732379A - Process for gathering and purifying panax japonicus total saponins by using D101 macroporous absorption resin - Google Patents

Abstract

The invention provides a process for gathering and purifying panax japonicus total saponins by using D101 macroporous absorption resin, which comprises the steps of: concentrating an extract of panax japonicus total saponins to 25.93mg/ml; filling a column with 95% ethanol by a wet method, wherein the diameter of the column is 15mm and the height of the column is 2220mm; balancing the volume of the column with distilled water to 33.5ml; absorbing 35 ml of 25.93mg/ml panax japonicus total saponin solution on the column at the speed of 1.85 multiplied column volume/ hour; after absorbing in static state for 1 hour, absorbing once again; eluting with distilled water till liquid flowing out from the column has no purple ring reaction; eluting the column with 50% ethanol with 17.11 multiplied column volume; recovering eluent, concentrating and drying the recovered eluent to obtain the product of the panax japonicus total saponins. Through test, the purity of the panax japonicus total saponins reaches 98.02% and the recovery rate reaches above 96.5%. Proved by experiments, the in vitro antioxidation effects of purified panax japonicus total saponins with different concentrations are all higher than that of unpurified panax japonicus total saponins, which shows that the gathering and purifying of the panax japonicus total saponins facilitates the improvement of the antioxidation activity of the panax japonicus total saponins.

Description

D101 enriching and purifying macroporous resin panax japonicus total saponins technology

Technical field

The present invention relates to functional food processing technique field, particularly D101 enriching and purifying macroporous resin panax japonicus total saponins technology.

Background technology

Rhizoma Panacis Japonici is the araliaceae ginseng plant, is used as medicine with its dry rhizome, claims Rhizoma Panacis japonici again, panax japonicus, Herba Rhodiolae Henryi etc.Rhizoma Panacis Japonici has the function of China north medicine Radix Ginseng and southern medicine Radix Notoginseng concurrently, is that Chinese Pharmacopoeia records kind, has effects such as strengthening by means of tonics, eliminating stasis to stop pain, hemostasis eliminate the phlegm.Pharmacological research shows, the contained saponins active substance of Rhizoma Panacis Japonici has extensive and firm pharmacological action to central nervous system, immune system and cardiovascular system, and [left side is sharp, Yuan Ding. the progress of Rhizoma Panacis Japonici chemical constituent and pharmacologically active [J]. the time precious traditional Chinese medical science traditional Chinese medicines 2005,16 (9): 838-841].Therefore, the extraction of panax japonicus total saponins and purification are the committed steps of exploitation Rhizoma Panacis Japonici product.

Adopt the chemical method gathering and purifying panax japonicus total saponins to have problems such as organic reagent is residual, environmental pollution, cost height.Also there is not report at present about D101 type enriching and purifying macroporous resin panax japonicus total saponins.The present invention explores the process conditions of having optimized D101 type enriching and purifying macroporous resin panax japonicus total saponins, for the exploitation of Rhizoma Panacis Japonici functional food lays the foundation.

Summary of the invention

Technical problem solved by the invention:, improved the panax japonicus total saponins purity and the higher response rate by D101 type enriching and purifying macroporous resin panax japonicus total saponins.

Concrete technical scheme of the present invention is as follows:

A kind of D101 enriching and purifying macroporous resin panax japonicus total saponins technology, its step comprises:

1), exsiccant Rhizoma Panacis Japonici section, place the reflux, extract, bottle, add 70% ethanol and press solid-liquid ratio and mix immersion 12 hour at 1: 12, heating and refluxing extraction 2 times is no less than 2 hours at every turn; Extracting liquid filtering merges, and is evaporated to the panax japonicus total saponins concentration of aqueous solution and is 25.9mg/ml, and is standby;

2), 95% ethanol wet method dress post, column diameter 15mm, high 220mm, distilled water balance columns volume is to 33.5ml;

3), with the panax japonicus total saponins aqueous solution 35ml that step 1) obtains, suitable 1.04 times of column volumes, with 1.85 times of column volumes/hour speed upper prop absorption, static absorption is heavily adsorbed after 1 hour 1 time;

4), the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction (molish reaction);

5), with 50% ethanol elution pillar of 17.11 times of column volumes, 1.25 times of column volumes of elution flow rate/hour, reclaim eluent;

6), the eluent that reclaims in the step 5) concentrated evaporate to dryness get the panax japonicus total saponins product, purity reaches 98.02% after measured, the response rate reaches more than 96.5%.

Ethanol % concentration refers to contained alcoholic acid milliliter number in 100 milliliters of ethanol waters among the present invention.

Description of drawings

Fig. 1 number is an abscissa with the test tube of auto partial sampler, and each manages saponin concentration is the elution curve that vertical coordinate is drawn

Remove the correlation curve of superoxide radical before and after Fig. 2 panax japonicus total saponins purification

By the visible best eluant of Fig. 1 elution curve is 50% ethanol.

As shown in Figure 2, the suppression ratio of panax japonicus total saponins strengthens with the concentration of total saponins, and the purified panax japonicus total saponins antioxidation in vitro effect of variable concentrations is all apparently higher than unpurified panax japonicus total saponins.The enriching and purifying that panax japonicus total saponins is described helps to strengthen its antioxidant activity effect.

The specific embodiment

Further specify technology contents of the present invention and effect below by example.

Embodiment 1

The preparation of Rhizoma Panacis Japonici saponin extract

Accurately take by weighing the exsiccant Rhizoma Panacis Japonici section of 3.0g, put in the round-bottomed bottle, the ethanol of adding 70% is pressed solid-liquid ratio and is mixed immersion 12 hours, heating and refluxing extraction secondary, each 2 hours at 1: 12; Extracting liquid filtering merges concentrating under reduced pressure; Obtain the concentrated solution of saponin concentration 25.9mg/ml.As supplying experimental liquid.

Embodiment 2

The selection of eluant

Step 1, new resin are cleaned the mechanical admixture of resin with distilled water, and sucking filtration anhydrates, and 95% soak with ethanol is 24 hours then, and is standby.

Step 2, take by weighing D101 type macroporous adsorbent resin after, with 95% ethanol wet method dress post, column diameter 15mm, high 220mm, distilled water balance columns volume is to 33.5ml;

2.24 times of column volume upper props of concentrated solution of the saponin concentration 25.9mg/ml that step 3, accurate absorption embodiment 1 obtain, preadsorption 1 hour, effluent heavily adsorb 1 time, collect 2.24 times of column volumes of effluent, flow velocity 2.33 column volumes/hour;

Step 4, use the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction;

Step 5, with 30% ethanol, 50% ethanol, 60% ethanol, 75% ethanol, each 3 times of column volume gradient elution of 95% ethanol, every 16.7ml are collected a pipe;

Step 6, mensuration are respectively managed the concentration of saponin in the eluent.

With the test tube of auto partial sampler number is abscissa, and each manages saponin concentration is that vertical coordinate is drawn elution curve.Elution curve is seen Fig. 1.Wherein 1～No. 6 pipe concentration is 30% ethanol, and 7～No. 12 pipe concentration is 50% ethanol, and 13～No. 18 pipe concentration is 60% ethanol, and 19～No. 24 pipe concentration is 70% ethanol, and concentration is 90% ethanol after No. 25 pipes.By the visible best eluant of elution curve is 50% ethanol.

Embodiment 3

D101 enriching and purifying macroporous resin panax japonicus total saponins

After taking by weighing of the method pretreatment of D101 type macroporous adsorbent resin by embodiment 2, with 95% ethanol wet method dress post, column diameter 15mm, high 400mm, distilled water balance columns volume is to 33.5ml; Accurate 2.56 times of column volume upper props of concentrated solution of drawing the saponin concentration 25.9mg/ml that embodiment 1 obtains, with 1.97 times of column volumes/hour the absorption of flow velocity upper prop, static absorption is heavily adsorbed after 1 hour 1 time; Earlier use the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction; Reuse 17.08 column volumes 50% ethanol, 1.81 column volumes/hour the flow velocity eluting, eluent concentrates evaporate to dryness and gets the panax japonicus total saponins product, measures panax japonicus total saponins purity 97.05%, the response rate 91.01%.

Embodiment 4

D101 enriching and purifying macroporous resin panax japonicus total saponins

After taking by weighing of the method pretreatment of D101 type macroporous adsorbent resin by embodiment 2, with 95% ethanol wet method dress post, column diameter 15mm, high 400mm, distilled water balance columns volume is to 33.5ml; Accurate 52.78 times of column volume upper props of concentrated solution of drawing the saponin concentration 25.9mg/ml that embodiment 1 obtains, with 1.97 times of column volumes/hour the absorption of flow velocity upper prop, static absorption is heavily adsorbed after 1 hour 1 time; Earlier use the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction; 50% ethanol of 18.41 times of column volumes of reuse, 1.33 times of column volumes/hour the flow velocity eluting, eluent concentrates evaporate to dryness and gets panax japonicus total saponins, measures panax japonicus total saponins purity 98.02%, the response rate 96.5%.

Embodiment 5

D101 enriching and purifying macroporous resin panax japonicus total saponins

After taking by weighing of the method pretreatment of D101 type macroporous adsorbent resin by embodiment 2, with 95% ethanol wet method dress post, column diameter 15mm, high 400mm, distilled water balance columns volume is to 33.5ml; Accurate 1.04 times of column volume upper props of concentrated solution of drawing the saponin concentration 25.9mg/ml that embodiment 1 obtains, with 1.97 times of column volumes/hour the absorption of flow velocity upper prop, static absorption is heavily adsorbed after 1 hour 1 time; Earlier use the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction; 50% ethanol of 17.11 times of column volumes of reuse, with 1.25 times of column volumes/hour the flow velocity eluting, eluent concentrates evaporate to dryness and gets the panax japonicus total saponins product, measures panax japonicus total saponins purity 98.18%, the response rate 97.10%.

Embodiment 6

The test of panax japonicus total saponins antioxidation in vitro

Employing pyrogallol autoxidation method mensuration [Cheng Chao, Zhu Yuchang, Mo Kaiju, Wang Xingping. the anti-oxidation characteristics research [J] of Bulbilus dioscoreae Saponin. Food Science 2007,28 (10): 83-86].

Step 1, accurately take by weighing panax japonicus total saponins before and after the purification respectively, the preparation distilled water solution, concentration is respectively 10mg/ml, 30mg/ml, 50mg/ml, 70mg/ml, 90mg/ml, and is standby as sample liquid;

Step 2, absorption 4.5mlpH8.250mmol/mlTris-HCI buffer, the 4.2ml distilled water mixes the back and be incubated 20 minutes in 25 ℃ of water-bath, sample liquid 1ml in adding respectively immediately after the taking-up; The 3mmol/L pyrogallol 0.3ml (with 10mmol/L HCl preparation) that adds 25 ℃ of preheatings more respectively, and replace pyrogallol HCl solution to make blank with 10mmol/L HCl solution;

Step 3, pour cuvette into after above-mentioned reactant liquor shaken up rapidly, under the 325nm wavelength, measured absorbance, calculate the value added of per minute absorbance in the linear range every 30 seconds;

Step 4, calculate panax japonicus total saponins suppression ratio before and after the purification respectively.

Suppression ratio (%)=(Δ A0-Δ A)/Δ A0 * 100%

In the formula, the autoxidation speed of Δ A0 pyrogallol; Δ A is the autoxidation speed of pyrogallol behind the adding saponin solution, and unit is the increment of absorbance per minute.

The result as shown in Figure 2.As shown in Figure 2, the suppression ratio of panax japonicus total saponins strengthens with the concentration of total saponins, and the purified panax japonicus total saponins antioxidation in vitro effect of variable concentrations is all apparently higher than unpurified panax japonicus total saponins.The enriching and purifying that panax japonicus total saponins is described helps to strengthen its antioxidant activity effect.

Claims (1)

1.D101 enriching and purifying macroporous resin panax japonicus total saponins technology is characterized in that, comprises the steps:

1), exsiccant Rhizoma Panacis Japonici section, place the reflux, extract, bottle, add 70% ethanol and press solid-liquid ratio and mix immersion 12 hour at 1: 12, heating and refluxing extraction 2 times is no less than 2 hours at every turn; Extracting liquid filtering merges, and is evaporated to the panax japonicus total saponins concentration of aqueous solution and is 25.9mg/ml, and is standby;

2), 95% ethanol wet method dress post, column diameter 15mm, high 220mm, distilled water balance columns volume is to 33.5ml;

3), with the panax japonicus total saponins aqueous solution 35ml that step 1) obtains, suitable 1.04 times of column volumes, with 1.85 column volumes/hour speed upper prop absorption, static absorption is heavily adsorbed after 1 hour 1 time;

4), the distilled water eluting, being washed till the pillar effluent does not have purple ring reaction;

5), with 50% ethanol elution pillar of 17.11 times of column volumes, elution flow rate 1.25 column volumes/hour, reclaim eluent;

6), the eluent that reclaims in the step 5) concentrated evaporate to dryness get the panax japonicus total saponins product, purity reaches 98.02% after measured, the response rate reaches more than 96.5%.

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