Summary of the invention
The object of the invention is, the preparation method of the fragrant pod acyl of a kind of 10-Catalpol is provided, it extracts the fragrant pod acyl of 10-Catalpol in Rhizoma Picrorhizae, improves extraction yield.
The present invention for achieving the above object, is achieved through the following technical solutions: the preparation method of the fragrant pod acyl of 10-Catalpol, comprises the steps:
1. get Rhizoma Picrorhizae pulverizing medicinal materials to 60 order, obtain medicinal material meal standby;
2. adopt former cotton gauze, make bilayer bag-shaped, the capacity of cotton yarn bag is splendid attire 5kg step medicinal material meal 1., obtains double-deck cotton yarn bag standby;
3. step medicinal material meal is 1. packed in step double-deck cotton yarn bag 2., every packed 3kg medicinal material meal that enters, by double-deck cotton yarn bag sealing, obtains medicinal material flour bag;
4. by step, the medicinal material meal bag in is 3. inserted in extraction vessel, 95% edible ethanol that divides 20 times of amounts that add medicinal material flour bag Chinese medicinal materials powdery thing weight for three times, add for the first time medicinal material meal weight 8 times of amounts, add respectively for the second time and for the third time 6 times of amounts of medicinal material meal weight, being heated to boiling reflux extracts respectively 2 hours, 1.5 hours, 1 hour, filter, merging filtrate obtains ethanol extract;
5. by step, the ethanol extract in 4. pumps in jacketed kettle by liquor pump, passes into ethylene glycol refrigerant circulation and be cooled to 0 ℃-4 ℃, standing 20-28 hour in interlayer;
6. the ethanol extract after step is standing in 5. pumps in plate-and-frame filter press, make ethanol extract under 19-22MPa pressure by sheet frame filtering layer, obtain clear and bright ethanol filtrate;
7. ethanol filtrate is inserted and in distiller, adds thermal distillation, to distillment be medicinal extract shape, obtain alcohol medicinal extract 300L;
8. alcohol medicinal extract is added to the chromatographic silica gel of 1/3 amount of medicinal material meal total amount, stir, be dried to moisture and separated silica gel moisture identical, be ground into 80 orders and obtain powdery to mix sample silica gel standby;
9. get 3.5kg100 order-200 order chromatographic silica gel, the eluent that adds ethyl acetate and ethanol to mix, stirs lower eliminating bubble, pours in chromatography column, the eluent of ethyl acetate and ethanol is eluted to repeatedly to bed surface no longer declines, cylinder is closely knit, the volume ratio of ethyl acetate and ethanol is 6:1 again;
10. by step, the powdery in is 8. mixed sample silica gel and is slowly added and on chromatography column, soak 9-11 hour;
carry out chromatography wash-out: the eluent wash-out mixing with ethyl acetate and ethanol, collect elutriant, distillating recovering solvent, it is yellow that distillment is, and when picking distillment and have " extension flag " phenomenon with sampling rod, be that the fragrant pod acyl of 50% 10-Catalpol is collected starting point, when distillment during without " extension flag " phenomenon for collecting terminal, the volume ratio of ethyl acetate and ethanol is 6:1;
ratio in the volume ratio 6:1 of ethyl acetate and ethanol is made elutriant, then will in elutriant, add step
after the heating activated carbon boiling of the fragrant pod acyl of 10-Catalpol 30% amount of middle collection, keep boiling 28-35 minute, filter, obtain the liquid that decolours;
to decolour liquid distillating recovering solvent to alcohol medicinal extract, then admix silica gel, repeating step 8., 9., 10.,
described content is carried out secondary chromatography, obtains elutriant and reclaims solvent to hypersaturated state, separates out the fragrant pod acyl of 10-Catalpol after letting cool.
The circulation temperature lowering to 3 ℃ of step of the present invention described in 5., time of repose is 24 hours.The pressure of step described in is 6. 20MPa.
Method of the present invention is in Rhizoma Picrorhizae, to extract the fragrant pod acyl of 10-Catalpol, the method packs 60 order Rhizoma Picrorhizae medicinal powders in the cotton yarn cloth bag of two bed thickness into, the difficult problems such as bumping, sticky pot and the sad filter of extracting solution have been solved in heat-processed, simultaneously, size and the explosive load of double-deck cotton yarn cloth bag have increased the flow of solvent between medical material bag, make composition be easy to diffusion; The method also adopts the stand at low temperature precipitation process of Rhizoma Picrorhizae powder ethanol extract, and processes through pressure filter, can remove most of cold alcohol insoluble matter macromolecular substance, has retained to greatest extent the fragrant pod acyl of 10-Catalpol, has improved the separation efficiency of column chromatography; The inventive method has also adopted ethyl acetate to carry out depickling, then by 5% sodium hydrogen carbonate solution equal-volume extraction ethyl acetate, remove again ethanol wherein, be combined into column chromatography eluent with ethanol again, solved the deficiency existing in available technology adopting chloroform-methanol technique.Meanwhile, in method of the present invention, to adopt ethanol be solvent and add decolouring with the gac that the fragrant pod acyl of 10-Catalpol equivalent is 30%, thereby can save mass energy, and avoided the fragrant pod acyl of 10-Catalpol because of the overheated deficiency of destroying, and further improved extraction yield.Adopt method of the present invention in Rhizoma Picrorhizae, to extract the fragrant pod acyl of 10-Catalpol, according to [assay] method detection under Rhizoma Picrorhizae item of < < Pharmacopoeia of People's Republic of China > > version in 2010, obtain the fragrant pod acyl of 10-Catalpol extraction yield and can reach 95%, purity reaches 92%.Method of the present invention is also easy to industrialization streamline and produces, can adopt the fragrant pod acyl of " extension flag " phenomenon judgement 10-catalpol content to surpass 50%, the content range that judges fast aborning the fragrant pod acyl of 10-Catalpol only needs 2 seconds kinds, thoroughly having solved the content that prior art detects the fragrant pod acyl of 10-Catalpol aborning needs several hours, although result accurately cannot instruct the deficiency of producing by rapid feedback.
Embodiment
The preparation method of the fragrant pod acyl of 10-of the present invention Catalpol is prepared according to following step:
1. get Rhizoma Picrorhizae pulverizing medicinal materials to 60 order, obtain medicinal material meal standby;
2. adopt former cotton gauze, make bilayer bag-shaped, the capacity of cotton yarn bag is splendid attire 5kg step medicinal material meal 1., obtains double-deck cotton yarn bag standby;
3. step medicinal material meal is 1. packed in step double-deck cotton yarn bag 2., every packed 3kg medicinal material meal that enters, by double-deck cotton yarn bag sealing, obtains medicinal material flour bag;
4. by step, the medicinal material meal bag in is 3. inserted in extraction vessel, 95% edible ethanol that divides 20 times of amounts that add medicinal material flour bag Chinese medicinal materials powdery thing weight for three times, add for the first time medicinal material meal weight 8 times of amounts, add respectively for the second time and for the third time 6 times of amounts of medicinal material meal weight, being heated to boiling reflux extracts respectively 2 hours, 1.5 hours, 1 hour, filter, merging filtrate obtains ethanol extract;
5. by step, the ethanol extract in 4. pumps in jacketed kettle by liquor pump, passes into ethylene glycol refrigerant circulation and be cooled to 0 ℃-4 ℃, standing 20-28 hour in interlayer;
6. the ethanol extract after step is standing in 5. pumps in plate-and-frame filter press, make ethanol extract under 19-22MPa pressure by sheet frame filtering layer, obtain clear and bright ethanol filtrate;
7. ethanol filtrate is inserted and in distiller, adds thermal distillation, to distillment be medicinal extract shape, obtain alcohol medicinal extract 300L;
8. alcohol medicinal extract is added to the chromatographic silica gel of 1/3 amount of medicinal material meal total amount, stir, be dried to moisture and separated silica gel moisture identical, be ground into 80 orders and obtain powdery to mix sample silica gel standby;
9. get 3.5kg100 order-200 order chromatographic silica gel, the eluent that adds ethyl acetate and ethanol to mix, stirs lower eliminating bubble, pours in chromatography column, the eluent of ethyl acetate and ethanol is eluted to repeatedly to bed surface no longer declines, cylinder is closely knit, the volume ratio of ethyl acetate and ethanol is 6:1 again;
10. by step, the powdery in is 8. mixed sample silica gel and is slowly added and on chromatography column, soak 9-11 hour;
carry out chromatography wash-out: the eluent wash-out mixing with ethyl acetate and ethanol, collect elutriant, distillating recovering solvent, it is yellow that distillment is, and when picking distillment and have " extension flag " phenomenon with sampling rod, be that the fragrant pod acyl of 50% 10-Catalpol is collected starting point, when distillment during without " extension flag " phenomenon for collecting terminal, the volume ratio of ethyl acetate and ethanol is 6:1;
ratio in the volume ratio 6:1 of ethyl acetate and ethanol is made elutriant, then will in elutriant, add step
after the heating activated carbon boiling of the fragrant pod acyl of 10-Catalpol 30% amount of middle collection, keep boiling 28-35 minute, filter, obtain the liquid that decolours;
to decolour liquid distillating recovering solvent to alcohol medicinal extract, then admix silica gel, repeating step 8., 9., 10.,
described content is carried out secondary chromatography, obtains elutriant and reclaims solvent to hypersaturated state, separates out the fragrant pod acyl of 10-Catalpol after letting cool.
The further scheme of the method for the invention is: the circulation temperature lowering to 3 ℃ of step described in 5., time of repose is 24 hours.
The preferred version of the method for the invention is: the pressure of step described in is 6. 20MPa.
The preparation method of the fragrant pod acyl of 10-of the present invention Catalpol, employing Tibet picrorhiza rhizome is raw material, its concrete steps are as follows:
1. get dry Tibet picrorhiza rhizome 150kg, be crushed to 60 orders;
2. adopt former cotton gauze, make bilayer bag-shaped, the capacity of cotton yarn bag is splendid attire 5kg step meal 1., obtains double-deck cotton yarn bag standby;
3. step medicinal material meal is 1. packed in step double-deck cotton yarn bag 2., every packed 3kg medicinal material meal that enters, by double-deck cotton yarn bag sealing, obtains medicinal material flour bag;
4. by step, the medicinal material flour bag in is 3. inserted in extraction vessel, 95% edible ethanol that divides 20 times of amounts that add raw material medicine bag traditional Chinese medicine powder shape thing weight for three times, add for the first time medicinal material meal weight 8 times of amounts, add respectively for the second time and for the third time 6 times of amounts of medicinal material meal weight, being heated to boiling reflux extracts respectively 2 hours, 1.5 hours, 1 hour, filter, merging filtrate obtains ethanol extract;
5. by step, the ethanol extract in 4. pumps in jacketed kettle by liquor pump, passes into ethylene glycol refrigerant circulation and is cooled to 3 ℃, standing 24 hours in interlayer;
6. the ethanol extract after step is standing in 5. pumps in plate-and-frame filter press, make ethanol extract under 20MPa pressure by sheet frame filtering layer, obtain clear and bright ethanol filtrate;
7. ethanol filtrate is inserted and in distiller, adds thermal distillation, to distillment be medicinal extract shape, obtain alcohol medicinal extract 300L;
8. get 3000ml alcohol medicinal extract, add 500g silica gel to mix thoroughly, dry to water content 2% identically with separated silica gel water content, be crushed to 80 orders, obtain pulverizing that to mix sample silica gel standby;
9. get 3.5kg200 order chromatographic silica gel, the eluent that adds ethyl acetate and ethanol to mix, stirs lower eliminating bubble, pours in chromatography column, the eluent of ethyl acetate and ethanol is eluted to repeatedly to bed surface no longer declines, cylinder is closely knit, the volume ratio of ethyl acetate and ethanol is 6:1 again;
10. by step, the powdery in is 8. mixed sample silica gel and is slowly added on chromatography column and soak 10 hours;
carry out chromatography wash-out: the eluent wash-out mixing with ethyl acetate and ethanol, collect elutriant, distillating recovering solvent, it is yellow that distillment is, and when picking distillment and have " extension flag " phenomenon with sampling rod, be that the fragrant pod acyl of 50% 10-Catalpol is collected starting point, when distillment during without " extension flag " phenomenon for collecting terminal, the volume ratio of ethyl acetate and ethanol is 6:1;
ratio in the volume ratio 6:1 of ethyl acetate and ethanol is made elutriant, then will in elutriant, add step
after the gac 40g ebuillition of heated of the fragrant pod acyl of 10-Catalpol 30% amount of middle collection, keep boiling 30 minutes, filter, obtain the liquid that decolours;
to decolour liquid distillating recovering solvent to alcohol medicinal extract, then admix silica gel, weight step 8., 9., 10.,
described content is carried out secondary chromatography, obtains elutriant and reclaims solvent to hypersaturated state, separates out the fragrant pod acyl of 10-Catalpol 50-60g after letting cool, and the purity of the fragrant pod acyl of 10-Catalpol is that the extraction yield of the fragrant pod acyl of 92%, 10-Catalpol is 95%.
The preparation method's of the fragrant pod acyl of 10-of the present invention Catalpol step
described in " extension flag " refer to that the distillment that sampling rod head picks reaches 6.5 * 10
11while mooring the viscosity factor value of/80 ℃, accompanyingly at sampling rod head and the distillment volume that picks, be greater than 2 times, sampling rod head more than volume, likeness in form flag shape.