CN101352487B - Long pepper extract, preparation method thereof, medicament composition and uses thereof - Google Patents

Abstract

The invention relates to a long pepper extract, a preparation method and a medicine combination thereof. The long pepper extract mainly contains methyl piperate and seven piperlongumine compounds, namely, peperlomguminine, piperine, decadiene amide, pepper nonine alkali, quinia, octadecyl triene amide and eicosatriene amide; by weight percentage, the total weight of the eight compounds accounts for 29.07-87.80 % of the total weight of the long pepper extract, wherein, peperlomguminine accounts for 0.95-3.43 % of the total weight of the long pepper extract, piperine accounts for 14.52-42.78 %, methyl piperate 0.82-3.11%, decadiene amide 1.36-4.27%, pepper nonine alkali 2.96-8.03%, quinia 1.04-3.26%, octadecyl triene amide 3.17-10.94% and eicosatriene amide 4.25-12.42%. The invention also provides the preparation method of the long pepper extract, the medicine combination and the application thereof.

Description

Fructus Piperis Longi extract, its preparation method, its pharmaceutical composition and application thereof

Technical field

The pharmaceutical composition and the application thereof that the present invention relates to a kind of Fructus Piperis Longi extract, its preparation method and comprise described Fructus Piperis Longi extract.

Background technology

Fructus Piperis Longi (Piper Longum L.) is the nearly maturation or the mature fruit of Piperaceae plant Fructus Piperis Longi.Fruit is gathered during by green blackening, removes impurity, dries.

The Fructus Piperis Longi function is warming spleen and stomach for dispelling cold, therapeutic method to keep the adverse QI flowing downwards pain relieving with curing mainly.Be used for institute's abdomen cold type of pain, migraine is had loose bowels in vomiting; The external treatment toothache.

Chemical constituent mainly contains other compositions such as amide alkaloid, volatile oil, acid, ester and lignanoid in the Fructus Piperis Longi.Wherein the more deep main effective ingredient of research is amide alkaloid, methyl piperate and volatile oil, wherein the amide alkaloid mainly plays a role at blood fat reducing, cardiovascular, convulsion, calmness, analgesia, aspect such as antibiotic, and volatile oil mainly plays a role at anti-gastric-ulcer, cardiovascular, antibiosis.

The amide alkaloid mainly contains piperine (piperine) in the Fructus Piperis Longi, the peaceful alkali of Fructus Piperis promise (piernonaline), the peaceful alkali of dehydrogenation Fructus Piperis promise (dehydropipernonaline), decadinene amide (N-isobutyldeca-trans-2-trans-4-dienamide), Piperlonguminine (piperlonguminine), dihydro Piperlonguminine (dehydropiperlonguminine), pipercide (pipercide), quinine element (guineensine), 18 carbon diene amide (N-isobutyl-2E, 4E-octadecadienamide), 18 carbon triolefin amide (N-Isobutyl-2E, 4E, 12Z-octadecatrienamide), 20 carbon diene amide (N-Isobutyl-2E, 4E-eicosadienamide), 20 carbon triolefin amide (N-Isobutyl-2E, 4E, 14Z-eicosatrienamide).

Acid mainly contains piperic acid (piperic acid) in the Fructus Piperis Longi; Ester mainly contains methyl piperate (mythyl piperate).

Modern pharmacological research proof Fructus Piperis Longi has the pharmacological action of following aspect:

1, anti-gastric-ulcer: can significantly suppress the formation of rat ulcer due to indometacin, dehydrated alcohol, aspirin, the acetic acid in advance the Fructus Piperis Longi ethanol extraction.Oral 0.25g/kg Fructus Piperis Longi ethanol extraction is respectively 62.9%, 36.9%, 38.2%, 52.8% to the suppression ratio of the above-mentioned moulding of rat ulcer.Rat ligation pylorus type gastric ulcer, gastric juice amount, total acidity gastric juice all there is remarkable inhibitory action.To reserpine type ulcer inhibition rate is 48.1%, shows that it has considerable influence to synthetic, the metabolism of prostaglandin E, and may be increases the epoxidase activity the synthetic and secretion of prostaglandin E is increased, and the gastric mucosa factor that is immune against attacks is destroyed.The gastrointestinal propulsion functions is not made significant difference.Can suppress its tension force and contractility to rabbit and rat ileum.

2, convulsion, calmness: rat ip150mg/kg has the convulsions effect of tangible anti-pentylenetetrazole.The convulsions rate is significantly reduced, electrofit and " audiogenic seizure " are also had obvious antagonism.Anticonvulsant action intensity can be compared with phenytoin Sodium and trimethadione.Injected in mice 25mg/kg, the movable number of times of 5min drops to for 244 times 95 times.With penthiobarbital synergism is arranged, the inhibitory action of antagonism morphine pentobarbital to breathing arranged, this antagonism is than pentylenetetrazole or nikethamide is strong and safety.

3, antiinflammatory sterilization: volatile oil all has inhibitory action to staphylococcus aureus, bacillus subtilis, Bacillus cereus, tubercule bacillus, dysentery bacterium, salmonella typhi T and B, yolk yellow eight folded bacterium etc. and influenza virus.Through 110 kinds of Chinese herbal medicine the antibacterial activity of Streptococcus mutans is measured discovery, Fructus Piperis Longi etc. have stronger antibacterial activity for 7 kinds.

4, to cardiovascular effect: to the Fructus Piperis Longi preparation that influences mice difference ip variable concentrations of mouse cardiac muscle trophism flow measurement, press the nutrient flow quantity measuring method behind the 30min, observe the picked-up ability of mouse cardiac muscle to 86Rb.The result increases myocardium picked-up ability to 86Rb with drug level and reduces gradually.Drug level is respectively 4.4g/kg, and uptake ratio is followed successively by 7.2 ± 4.000} (P＜0.01), 6.11 ± 4.21%CP＜0.05 when 4.9g/kg and 11.1g/kg) and 3.3 ± 4.20%＜P＞0.01).When drug dose increases to 17.8g/kg, opposite with The above results, can make the mouse cardiac muscle uptake ratio be increased to 9.5 ± 3.68% (P＜0.01), the effect under this dosage is equivalent to order one receptor stimulating agent, can increase coronary flow, improves myocardial metabolism.Therefore, Fructus Piperis Longi can be respectively as β one receptor blocking agent and β one receptor stimulating agent under various dose.

5, anoxia enduring, anti-acute myocardial ischemia, coronary dilating, anti-arrhythmia: Fructus Piperis Longi volatile oil can improve the mice hypoxia-bearing capability under the reduced pressure, and medication group survival rate is 93.1%, and matched group is 18.6%.The normal pressure hypoxia endurance test shows, can prolong the time-to-live 37.8% of mice, improves the mice life span that the oxygen consumption due to different third adrenal gland's rope increases, and improves 30% than matched group, and sodium nitrite is caused the anoxia mice, improves 64% life span.Prolong the death time of potassium cyanide poisoning mice, improve 66.6% with comparing.Folder is closed the histanoxia death time that the mice bilateral carotid causes, compare, can prolong 58.3% with matched group.Significantly antagonism is changed and the decreased heart rate effect by the rabbit T ripple that pituitrin causes.Volatile oil ig has the arrhythmia effect due to remarkable prevention chloroform one epinephrine; But oral nothing obviously prevents chloroform to bring out the ventricular fibrillation effect, but has arrhythmia effect due to the anti-barium chloride with the propranolol highly significant; Can prolong ventricular premature contraction due to the aconitine, ventricular tachycardia and chamber property fibrillation and asystole time; Can not significantly improve guinea pig heart to the ouabain tolerance of poisoning, but certain increase trend is arranged, and significantly improve the lethal tolerance of heart.

6, effect for reducing blood fat: early eighties, Wurenbayar is by a large amount of zooperies, from 11 flavor mongolian medicine medical materials, found medical herbs simply, and found that other 10 flavor does not all have obvious drug effect, i.e. Piperaceae plant Fructus Piperis Longi (Piper Longum L.) with obvious effect for reducing blood fat.The curative effect highly significant of the extract Ge Genqin of Fructus Piperis Longi, active princlple are piperlongumine alkaloid and methyl piperate.

7, other effect: piperlongumine 0.1mg/kg has the effect of obvious reduction dog blood pressure; Antifertility activity to female rat or rat is 0%, with its fruit powder mixes in feedstuff, laboratory animal is had obvious effect.

At present the research of Fructus Piperis Longi effective ingredient effect for reducing blood fat aspect is mainly concentrated on and extract effective site, effective component extracting and effective ingredient carried out aspect such as structure of modification and carry out, but embody the blood fat reducing of mongolian medicine Fructus Piperis Longi, atherosclerosis, resist myocardial ischemia and still extract and purify natural piperlongumine alkaloid and the methyl piperate of other many-sided protective effects such as cardiovascular mixed the effect of effective site the most comprehensive and the most satisfactory.

At present Fructus Piperis Longi extract uses ethanol extraction usually, and separation method such as normal pressure or decompression and solvent recovery etc. by routine separate, and carry out purification by several different methods such as saponification, column chromatography and high pressure lipuid chromatography (HPLC).But it is low few with active component to extract the Fructus Piperis Longi extract active constituent content that obtains.

Disclose among CN 1322543 A Fructus Piperis Longi has been carried out ethanol extraction, and by the saponification alkaloidal method of piperlongumine of purifying, the shortcoming of this method is: (1) has used poisonous and carcinogenic solvent benzol in saponification, can cause the environmental pollution in the production, can cause that the direct labor poisons, can bring toxic solvent residual, bring safety issue to product; (2) this method scientific and technological content is lower, and the extract active constituent content is also low to have only 6-8%, thereby the effect of product just can not be very good, and need take a large amount of extracts, just is more prone to cause safety issue.

In other document, disclose Fructus Piperis Longi carried out ethanol extraction, and carried out purification by column chromatography or high pressure lipuid chromatography (HPLC), they all be the laboratory purification with separate Fructus Piperis Longi in the method for the individualized compound of amide alkaloid.Shortcoming is: (1) silica gel, alumina column chromatography and high pressure lipuid chromatography (HPLC) are difficult for industrialization; (2) use silica gel and aluminium oxide to have environmental issue in a large number; (3) product cost height; (4) be difficult to obtain than the alkaloidal effective site of multiamide.

Summary of the invention

The object of the invention is to provide a kind of Fructus Piperis Longi extract.

Second purpose of the present invention is to provide the pharmaceutical composition that comprises above-mentioned Fructus Piperis Longi extract.

The 3rd purpose of the present invention is to provide the preparation method of above-mentioned Fructus Piperis Longi extract.

The 4th purpose of the present invention is to provide above-mentioned Fructus Piperis Longi extract or pharmaceutical composition to be used to prepare the purposes of medicine, and wherein said medicine is used for the treatment of hyperlipidemia, analgesia, antiinflammatory or gastric ulcer.

A kind of Fructus Piperis Longi extract provided by the invention, mainly contain methyl piperate (mythyl piperate) and seven kinds of piperlongumine chemical compounds and methyl piperate: Piperlonguminine (piperlonguminine), piperine (piperine), decadinene amide (N-isobutyldeca-trans-2-trans-4-dienamide), the peaceful alkali of Fructus Piperis promise (piernonaline), quinine element (guineensine), 18 carbon triolefin amide (N-Isobutyl-2E, 4E, 12Z-octadecatrienamide) with 20 carbon triolefin amide (N-Isobutyl-2E, 4E, 14Z-eicosatrienamide) eight chemical compounds, by weight percentage, eight kinds of chemical compound gross weights account for 29.07～87.80% of Fructus Piperis Longi extract gross weight, wherein Piperlonguminine accounts for 0.95～3.43% of Fructus Piperis Longi extract gross weight, piperine accounts for 14.52～42.78% of Fructus Piperis Longi extract gross weight, methyl piperate accounts for 0.82～3.11% of Fructus Piperis Longi extract gross weight, the decadinene amide accounts for 1.36～4.27% of Fructus Piperis Longi extract gross weight, the peaceful alkali of Fructus Piperis promise accounts for 2.96～8.03% of Fructus Piperis Longi extract gross weight, the quinine element accounts for 1.04～3.26% of Fructus Piperis Longi extract gross weight, 3.17～10.94%, the 20 carbon triolefin amide that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight account for 4.25～12.42% of Fructus Piperis Longi extract gross weight.

Above-mentioned Fructus Piperis Longi extract of the present invention, by weight percentage, wherein seven kinds of piperlongumine chemical compounds and methyl piperate totally eight kinds of chemical compound gross weights preferably account for 49.64～71.89% of Fructus Piperis Longi extract gross weight, wherein Piperlonguminine preferably accounts for 1.88～2.54% of Fructus Piperis Longi extract gross weight, piperine preferably accounts for 24.16～35.16% of Fructus Piperis Longi extract gross weight, methyl piperate preferably accounts for 1.72～2.35% of Fructus Piperis Longi extract gross weight, the decadinene amide preferably accounts for 2.21～2.67% of Fructus Piperis Longi extract gross weight, the peaceful alkali of Fructus Piperis promise preferably accounts for 4.58～6.75% of Fructus Piperis Longi extract gross weight, the quinine element preferably accounts for 1.83～2.44% of Fructus Piperis Longi extract gross weight, 6.05～9.43%, the 20 carbon triolefin amide that 18 carbon triolefin amide preferably account for the Fructus Piperis Longi extract gross weight preferably account for 7.21～10.55% of Fructus Piperis Longi extract gross weight.

The present invention also provides and comprises the pharmaceutical composition of Fructus Piperis Longi extract as mentioned above.

The pharmaceutical composition of described Fructus Piperis Longi extract is the pharmaceutical preparation of oral administration, coelenteron administration, spray delivery, ejection preparation, infusion preparation or Transdermal absorption.

The pharmaceutical composition of described Fructus Piperis Longi extract, wherein, the pharmaceutical preparation of oral administration is selected from a kind of in tablet, capsule, powder, granule, crystal, solution, suspension, soup, syrup, elixir, tea and the masticatory.

In the described pharmaceutical composition, described Fructus Piperis Longi extract can use separately, perhaps and well known to a person skilled in the art that similar active composition unites use.In addition, pharmaceutical composition of the present invention also can comprise pharmaceutically acceptable carrier.These carriers are well known to a person skilled in the art.

In the embodiment of the present invention aspect this, described pharmaceutical composition is mixed with the pharmaceutical preparation of oral administration, coelenteron administration, spray delivery, ejection preparation, infusion preparation and Transdermal absorption.In a preferred embodiment, the pharmaceutical preparation of described oral administration is selected from tablet, capsule, powder, granule, crystal, solution, suspension, soup, syrup, elixir, tea and masticatory.These pharmaceutical preparatioies all can utilize the technology of well known to a person skilled in the art to prepare.

A kind of preparation provided by the invention is the method for Fructus Piperis Longi extract as mentioned above, and this method comprises the following steps:

Step (a) is 70%～100% (v/v) ethanol extraction with Fructus Piperis Longi concentration, to obtain ethanol extract;

Step (b) is with the ethanol extract decompression recycling ethanol, to obtain fluid extract;

Step (c) is placed fluid extract and was separated out crystallization in 24～96 hours;

Step (d) will separate out after the crystallization fluid extract with 4～10 times (w: 10～20% (w/w) KOH aqueous solution saponification w), to obtain the Fructus Piperis Longi non-saponifiable matter;

Step (e) with Fructus Piperis Longi non-saponifiable matter condition is: pressure=9～32MPa, flow velocity=2～6mL/min, Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL=1: 1～1: 4, temperature is that 35～77 ℃ CO2 supercritical rectification method carries out separation and purification, obtains described Fructus Piperis Longi extract.

The preparation method of Fructus Piperis Longi extract as mentioned above, wherein,

Used concentration of alcohol is 95% (v/v) in the step (a);

Placing in the step (c) and separating out the crystalline time is 48 hours;

Used KOH concentration is 15% (w/w) in the step (d), and the KOH consumption is 6 times of (w: w);

CO2 supercritical rectification method process conditions are in the step (e): pressure=11.5MPa, and flow velocity=3mL/min, Fructus Piperis Longi extract quality g: ethanol volume mL=1: 2, thermograde=35-49-63-77 ℃.

Beneficial effect of the present invention mainly comprises:

1, Fructus Piperis Longi extract active constituent content height provided by the invention and active component comprise many.The blood fat reducing effective ingredient is amide alkaloid and methyl piperate in the Fructus Piperis Longi, the amide alkaloid mainly contains piperine (piperine), the peaceful alkali of Fructus Piperis promise (piernonaline), the peaceful alkali of dehydrogenation Fructus Piperis promise (dehydropipernonaline), decadinene amide (N-isobutyldeca-trans-2-trans-4-dienamide), Piperlonguminine (piperlonguminine), dihydro Piperlonguminine (dehydropiperlonguminine), pipercide (pipercide), quinine element (guineensine), 18 carbon diene amide (N-isobutyl-2E, 4E-octadecadienamide), 18 carbon triolefin amide (N-Isobutyl-2E, 4E, 12Z-octadecatrienamide), 20 carbon diene amide (N-Isobutyl-2E, 4E-eicosadienamide), 20 carbon triolefin amide (N-Isobutyl-2E, 4E, 14Z-eicosatrienamide).

The present invention's controlled condition after alcohol extraction, saponification is carried out supercritical extraction and purification, and efficiently purifying concentrates active princlple content when keeping piperlongumine alkaloid all the components and methyl piperate to greatest extent.Therefore, in Fructus Piperis Longi extract of the present invention, it contains Piperlonguminine (piperlonguminine), piperine (piperine), methyl piperate (mythylpiperate), decadinene amide (N-isobutyldeca-trans-2-trans-4-dienamide), the peaceful alkali of Fructus Piperis promise (piernonaline), quinine element (guineensine), 18 carbon triolefin amide (N-Isobutyl-2E, 4E, 12Z-octadecatrienamide) and 20 carbon triolefin amide (N-Isobutyl-2E, 4E, 14Z-eicosatrienamide) eight chemical compounds, the percentage by weight that eight kinds of chemical compound gross weights account for the Fructus Piperis Longi extract gross weight is up to 87.80%, wherein the Piperlonguminine percentage by weight that accounts for the Fructus Piperis Longi extract gross weight is up to 3.43%, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is up to 42.78%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is up to 3.11%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is up to 4.27%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is up to 8.03%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is up to 3.26%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is up to 10.94%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is up to 12.42%.

2, the present invention uses the KOH aqueous solution as saponifier, avoids with an organic solvent bringing dissolvent residual and toxicity.

3, the present invention can make the abundant enrichment of active princlple by this green purifying process of CO2 supercritical rectification method; organic solvent residual, heavy-metal residual or environmental issue that the traditional purification process of avoiding is again simultaneously brought have guaranteed the unification of security of products, effectiveness and environmental conservation.

Description of drawings

Fig. 1 is Piperlonguminine, piperine and methyl piperate standard reference material HPLC collection of illustrative plates;

Fig. 2 is decadinene amide, the peaceful alkali of Fructus Piperis promise, quinine element, 18 carbon triolefin amide and 20 carbon triolefin amide standard reference material HPLC collection of illustrative plates;

Fig. 3 is a Fructus Piperis Longi extract HPLC collection of illustrative plates;

Fig. 4 is a CO2 supercritical rectification separation and purification Fructus Piperis Longi extract flow chart.

The specific embodiment

Below by specific embodiment, the present invention will be further described.

It should be appreciated by those skilled in the art that these embodiment just are used for illustration purpose, and do not limit the scope of the invention.Protection scope of the present invention only is subjected to the restriction of claims.Under the condition that does not deviate from claims scope, those skilled in the art can carry out various modifications and improvement to various aspects of the present invention, and these modifications and improvement also belong to protection scope of the present invention.

In addition, unless it should be noted that and specialize, below among the embodiment used various materials and reagent all be material and reagent commonly used in this area, can obtain by conventional commercial sources; Method therefor is and well known to a person skilled in the art conventional method.In addition, unless specialize, the percentage composition of solution is a volumn concentration, i.e. % (v/v), and multiple is the weight multiple, the percentage composition of other material is w/w percentage ratio, i.e. % (w/w).

Embodiment 1

Take by weighing Fructus Piperis Longi 7.37kg (available from the Hebei Anguo), 20 orders are pulverized, and add 70% ethanol 58.96L, soaked overnight, stirring at room was extracted 4 hours, sucking filtration, residue after extracting for the first time carries out the second time respectively and extracts, every part adds 70% ethanol 44.22L, and stirring at room is extracted 2h, sucking filtration, merge extracted twice filtrate, be evaporated to driedly, obtain extractum (A), 880g weighs; Extractum (A), place, separate out the precipitate piperine, sucking filtration, remove precipitate, mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain Fructus Piperis Longi ethanol extraction extractum (B) 750g, and carry out assay, eight kinds of effective ingredient total contents 14.11% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Preparation 10%KOH solution 2.2L, take by weighing ethanol extraction extractum 550g (content is 14.11%), add the KOH solution for preparing, jolting 30 minutes was placed 3 hours, emitted lower floor's alkali liquor, upper strata (not saponification layer) solution adds no saline 2.5L, half an hour is placed in jolting 5 minutes, bleeds off water layer, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with upper strata grease, and with 95% ethanol wash clean, be evaporated to dried, non-saponifiable matter extractum weigh (C) 420.5g, and carry out assay, as a result eight kinds of effective ingredient total contents 21.53% of non-saponifiable matter extractum (C).

Use supercritical rectification process (seeing accompanying drawing 4) to carry out supercritical rectification and separate purification, concrete operations step and condition are as follows: get non-saponifiable matter (C) 420.5g, add 4 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=9 (MPa), flow velocity=6mL/min, and concentration=1: 4 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-39-59-77 ℃.Experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.After removing ethanol, decompression obtains Fructus Piperis Longi extract (D) 252.8g.Eight kinds of chemical compound gross weights of assay account for 29.07% of Fructus Piperis Longi extract gross weight, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 0.95% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 14.52%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 0.82%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 1.36%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 2.96%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 1.04%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is that the percentage by weight that 3.17%, two ten carbon triolefin amide accounts for the Fructus Piperis Longi extract gross weight is 4.25%.

Method for measuring and analyzing content (seeing accompanying drawing 1,2,3): all assays all use the middle external standard method of high performance liquid chromatography (HPLC) to measure in this patent, used instrument is a high performance liquid chromatograph, the Agilent1100 pump, 1100 diode array detectors, 1100 automatic samplers, the Agilent chromatographic work station.

Totally eight of used reference substances, Piperlonguminine (lot number: 20031110) wherein, methyl piperate (lot number: 20040202), decadinene amide (lot number: 20040107), the peaceful alkali of Fructus Piperis promise (lot number: 20040227), the quinine element (lot number: 20040115), 18 carbon triolefin amide (lot number: 20040321) and 20 carbon triolefin amide (lot number: 20040322) above seven for the self-control; Another piperine reference substance is bought lot number from Nat'l Pharmaceutical ﹠ Biological Products Control Institute: 0775-9702, assay is used.

Embodiment 2

Take by weighing Fructus Piperis Longi 7.37kg (available from the Hebei Anguo), 20 orders are pulverized, and add 95% ethanol 58.96L, soaked overnight, stirring at room was extracted 4 hours, sucking filtration, residue after extracting for the first time carries out the second time respectively and extracts, every part adds 95% ethanol 44.22L, and stirring at room is extracted 2h, sucking filtration, merge extracted twice filtrate, be evaporated to driedly, obtain extractum (A), 662g weighs; Extractum (A), place, separate out the precipitate piperine, sucking filtration, remove precipitate, mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain Fructus Piperis Longi ethanol extraction extractum (B) 556g, and carry out assay, eight kinds of effective ingredient total contents 22.45% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Preparation 8%KOH solution 3.3L, take by weighing ethanol extraction extractum 556g (content is 23.11%), add the KOH solution for preparing, jolting 30 minutes was placed 3 hours, emitted lower floor's alkali liquor, upper strata (not saponification layer) solution adds no saline 2.5L, half an hour is placed in jolting 5 minutes, bleeds off water layer, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with upper strata grease, and with 95% ethanol wash clean, be evaporated to dried, non-saponifiable matter extractum weigh (C) 305.6g, and carry out assay, as a result eight kinds of effective ingredient total contents 29.89% of non-saponifiable matter extractum (C).

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter (C) 305.6g, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=28 (MPa), flow velocity=2mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-45-59-71 ℃.Experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.After removing ethanol, decompression obtains Fructus Piperis Longi extract (D) 132.6g.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 39.77%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.49% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 19.52%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.04%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 1.78%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 3.86%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 1.46%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is that the percentage by weight that 4.77%, two ten carbon triolefin amide accounts for the Fructus Piperis Longi extract gross weight is 5.85%.

Embodiment 3

Take by weighing Fructus Piperis Longi 7.37kg (available from the Hebei Anguo), 20 orders are pulverized, and add 100% ethanol 58.96L, soaked overnight, stirring at room was extracted 4 hours, sucking filtration, residue after extracting for the first time carries out the second time respectively and extracts, every part adds 100% ethanol 44.22L, and stirring at room is extracted 2h, sucking filtration, merge extracted twice filtrate, be evaporated to driedly, obtain extractum (A), 650g weighs; Extractum (A), place, separate out the precipitate piperine, sucking filtration, remove precipitate, mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain Fructus Piperis Longi ethanol extraction extractum (B) 550g, and carry out assay, eight kinds of effective ingredient total contents 20.67% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Preparation 20%KOH solution 5.5L, take by weighing ethanol extraction extractum 550g (content is 20.67%), add the KOH solution for preparing, jolting 30 minutes was placed 3 hours, emitted lower floor's alkali liquor, upper strata (not saponification layer) solution adds no saline 2.5L, half an hour is placed in jolting 5 minutes, bleeds off water layer, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with upper strata grease, and with 95% ethanol wash clean, be evaporated to dried, non-saponifiable matter extractum weigh (C) 320.5g, and carry out assay, as a result eight kinds of effective ingredient total contents 30.65% of non-saponifiable matter extractum (C).

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter (C) 320.5g, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=32 (MPa), flow velocity=4mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-45-59-71 ℃.Experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.After removing ethanol, decompression obtains Fructus Piperis Longi extract (D) 141.3g.Eight kinds of chemical compound gross weights of assay account for 41.65% of Fructus Piperis Longi extract gross weight, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.61% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 20.15%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.23%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.04%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 3.94%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 1.75%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is that the percentage by weight that 5.06%, two ten carbon triolefin amide accounts for the Fructus Piperis Longi extract gross weight is 5.87%.

Embodiment 4

Take by weighing Fructus Piperis Longi 7.37kg (available from the Hebei Anguo), 20 orders are pulverized, and add 95% ethanol 58.96L, soaked overnight, stirring at room was extracted 4 hours, sucking filtration, residue after extracting for the first time carries out the second time respectively and extracts, every part adds 95% ethanol 44.22L, and stirring at room is extracted 2h, sucking filtration, merge extracted twice filtrate, be evaporated to driedly, obtain extractum (A), 680g weighs; Extractum (A), place, separate out the precipitate piperine, sucking filtration, remove precipitate, mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain Fructus Piperis Longi ethanol extraction extractum (B) 540g, and carry out assay, eight kinds of effective ingredient total contents 23.11% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Preparation 10%KOH solution 5.5L, take by weighing ethanol extraction extractum 540g (content is 23.11%), add the KOH solution for preparing, jolting 30 minutes was placed 3 hours, emitted lower floor's alkali liquor, upper strata (not saponification layer) solution adds no saline 5L, half an hour is placed in jolting 5 minutes, bleeds off water layer, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with upper strata grease, and with 95% ethanol wash clean, be evaporated to dried, non-saponifiable matter extractum weigh (C) 242.5g, and carry out assay, as a result eight kinds of effective ingredient total contents 36.42% of non-saponifiable matter extractum (C).

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter (C) 242.5g, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=19.5 (MPa), flow velocity=4mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-43-51-71 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.After removing ethanol, decompression obtains Fructus Piperis Longi extract (D) 162.8g.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 49.64%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.88% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 24.16%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.72%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.21%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 4.58%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 1.83%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is that the percentage by weight that 6.05%, two ten carbon triolefin amide accounts for the Fructus Piperis Longi extract gross weight is 7.21%.

Embodiment 5

Take by weighing Fructus Piperis Longi 7.37kg (available from the Hebei Anguo), 20 orders are pulverized, and add 95% ethanol 58.96L, soaked overnight, stirring at room was extracted 4 hours, sucking filtration, residue after extracting for the first time carries out the second time respectively and extracts, every part adds 95% ethanol 44.22L, and stirring at room is extracted 2h, sucking filtration, merge extracted twice filtrate, be evaporated to driedly, obtain extractum (A), 655g weighs; Extractum (A), place, separate out the precipitate piperine, sucking filtration, remove precipitate, mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain Fructus Piperis Longi ethanol extraction extractum (B) 546g, and carry out assay, eight kinds of effective ingredient total contents 24.03% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Preparation 15%KOH solution 5.5L, take by weighing ethanol extraction extractum 546g (content is 24.03%), add the KOH solution for preparing, jolting 30 minutes was placed 3 hours, emitted lower floor's alkali liquor, upper strata (not saponification layer) solution adds no saline 5L, half an hour is placed in jolting 5 minutes, bleeds off water layer, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with upper strata grease, and with 95% ethanol wash clean, be evaporated to dried, non-saponifiable matter extractum weigh (C) 242.5g, and carry out assay, as a result eight kinds of effective ingredient total contents 36.42% of non-saponifiable matter extractum (C).

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter (C) 242.5g, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=19 (MPa), flow velocity=4mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-43-51-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.After removing ethanol, decompression obtains Fructus Piperis Longi extract (D) 162.8g.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 51.73%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.15% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 24.32%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.94%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.56%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 4.84%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 2.06%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 6.41%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 7.45%.

Embodiment 6

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amounts (1600L), 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse (1200L), 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in about 2500L reclaim under reduced pressure filtrate obtains extract extractum (A) 25.5kg.

Extractum (A), placed 48 hours, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1.0kg, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 14kg, and carry out assay, eight kinds of effective ingredient total contents 34.28% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 14kg, the KOH aqueous solution that adds 6 times of amount (84kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 7.3kg.And carry out assay, eight kinds of effective ingredient total contents 53.36% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 7.3kg, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=11.5 (MPa), flow velocity=3mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-49-63-77 ℃.Experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 5.85kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 61.04%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.22% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 29.63%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.83%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.40%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 5.90%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 1.99%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 8.09%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 8.98%.

Embodiment 7

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 27.3kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1847g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 18kg, and carry out assay, eight kinds of effective ingredient total contents 28.02% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 18kg, the KOH aqueous solution that adds 6 times of amount (108kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 7.9kg.And carry out assay, eight kinds of effective ingredient total contents 49.56% of non-saponifiable matter extractum (C) as a result.

Use the supercritical rectification process (seeing accompanying drawing 4) that has four joint rectifying columns of development voluntarily to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 7.9kg, add 2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=11.5 (MPa), flow velocity=3mL/min, and concentration=1: 2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-49-63-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 6.11kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 59.99%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.28% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 28.25%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 1.87%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.27%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 5.86%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 2.04%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 8.19%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 9.23%.

Embodiment 8

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 26.1kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1869g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 17.4kg, and carry out assay, eight kinds of effective ingredient total contents 27.45% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 17.4kg, the KOH aqueous solution that adds 6 times of amounts 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 6.9kg.And carry out assay, eight kinds of effective ingredient total contents 48.26% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 6.9kg, add 1.5 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=19.5 (MPa), flow velocity=3mL/min, and concentration=1: 1.5 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=41-51-63-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 4.22kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 69.86%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.50% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 34.07%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 2.31%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.60%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 6.62%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 2.36%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 9.22%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 10.18%.

Embodiment 9

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 24.8kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1641g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 16.7kg, and carry out assay, eight kinds of effective ingredient total contents 29.33% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 16.7kg, the KOH aqueous solution that adds 6 times of amount (100kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 6.2kg.And carry out assay, eight kinds of effective ingredient total contents 50.39% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 6.2kg, add 1.5 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=19.5 (MPa), flow velocity=3mL/min, and concentration=1: 1.5 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=43-51-63-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 4.26kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 71.89%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.54% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 35.16%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 2.35%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 2.67%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 6.75%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 2.44%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 9.43%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 10.55%.

Embodiment 10

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 22.8kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 2013g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 15.8kg, and carry out assay, eight kinds of effective ingredient total contents 26.34% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 15.8kg, the KOH aqueous solution that adds 6 times of amount (95kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 6.1kg.And carry out assay, eight kinds of effective ingredient total contents 47.28% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 6.1kg, add 1.2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=23 (MPa), flow velocity=2.5mL/min, and concentration=1: 1.2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-49-63-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 3.15kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 79.75%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.88% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 38.85%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 2.81%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 3.59%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 7.41%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 2.99%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 9.98%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 11.24%.

Embodiment 11

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 27.9kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1947g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 18.7kg, and carry out assay, eight kinds of effective ingredient total contents 28.43% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 18.7kg, the KOH aqueous solution that adds 6 times of amount (112kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 8.2kg.And carry out assay, eight kinds of effective ingredient total contents 47.56% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 8.2kg, add 1.2 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=23 (MPa), flow velocity=2.0mL/min, and concentration=1: 1.2 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=35-49-63-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 2.91kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 81.39%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.96% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 39.14%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 2.89%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 3.77%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 7.65%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 3.02%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 10.33%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 11.63%.

Embodiment 12

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 26.25kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 1539g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 15.2kg, and carry out assay, eight kinds of effective ingredient total contents 27.47% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 15.2kg, the KOH aqueous solution that adds 6 times of amount (91.2kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 5.6kg.And carry out assay, eight kinds of effective ingredient total contents 46.55% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 5.6kg, add 1.0 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=27 (MPa), flow velocity=2.5mL/min, and concentration=1: 1.0 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=41-59-69-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 1.66kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 87.80%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 3.26% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 42.78%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 3.11%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 4.21%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 8.03%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 3.26%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 10.85%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 12.30%.

Embodiment 13

Get 200kg Fructus Piperis Longi (purchasing) in the Hebei Anguo, pulverize by (FFC-330 type melon type pulverizer), 20 orders sieve, with 8 times of amount 95% soak with ethanol 24h, stirring at normal temperature 4h, filter, reclaim ethanol, 6 times of amounts of filtering residue reuse, 95% ethanol extraction for the second time, stirring at normal temperature 2h, filter, merge first and second time filtrate, the ethanol in the reclaim under reduced pressure filtrate obtains extract extractum (A) 22.3kg.

Extractum (A), place, separate out the precipitate piperine, sucking filtration, obtaining precipitate is 2159g, and mother solution is flung to residual ethanol to there not being the alcohol flavor in water-bath, obtain ethanol extraction extractum (B) 14.8kg, and carry out assay, eight kinds of effective ingredient total contents 25.98% of Fructus Piperis Longi ethanol extraction extractum (B) as a result.

Get ethanol extraction extractum (B) 14.8kg, the KOH aqueous solution that adds 6 times of amount (89kg) 15% behind the stirring 10min, is emitted saponification liquor in detached dowel, placement is spent the night, after discard alkali liquor, upper strata (not saponification layer) solution adds pure water 50L, stirs 10 minutes, after placement is spent the night, emit water layer, non-saponifiable matter is put into agitated kettle, adds pure water 50L, stirred 10 minutes, placed 30 minutes, and divided water-yielding stratum, 4-6 time repeatedly, non-saponifiable matter is washed till neutrality, with 95% ethanol non-saponifiable matter in the agitated kettle is cleaned, merged the decompression of ethanol liquid and non-saponifiable matter and steam ethanol and moisture, obtain non-saponifiable matter extractum (C) 5.1kg.And carry out assay, eight kinds of effective ingredient total contents 49.24% of non-saponifiable matter extractum (C) as a result.

Use supercritical rectification process (seeing accompanying drawing 4) to carry out the supercritical rectification and purification, concrete operations step and condition are as follows: get non-saponifiable matter extractum (C) 5.1kg, add 1.0 times of amount dehydrated alcohol, the back sample introduction that stirs carries out separation and purification, pressure=32 (MPa), flow velocity=2.0mL/min, and concentration=1: 1.0 (Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL), thermograde=41-59-69-77 ℃.Each experiment finishes the back with 800mL absolute ethanol washing dephlegmator, and wash solution merges in the product.Decompression removes ethanol and obtains Fructus Piperis Longi extract (D) 1.72kg.The percentage by weight that eight kinds of chemical compound gross weights of assay account for the Fructus Piperis Longi extract gross weight is 87.14%, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 3.43% to Piperlonguminine, the percentage by weight that piperine accounts for the Fructus Piperis Longi extract gross weight is 42.20%, the percentage by weight that methyl piperate accounts for the Fructus Piperis Longi extract gross weight is 3.01%, the percentage by weight that the decadinene amide accounts for the Fructus Piperis Longi extract gross weight is 3.99%, the percentage by weight that the peaceful alkali of Fructus Piperis promise accounts for the Fructus Piperis Longi extract gross weight is 7.94%, the percentage by weight that the quinine element accounts for the Fructus Piperis Longi extract gross weight is 3.21%, the percentage by weight that 18 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 10.94%, and the percentage by weight that 20 carbon triolefin amide account for the Fructus Piperis Longi extract gross weight is 12.42%.

Embodiment 14

The effective site of Chinese medicine Fructus Piperis Longi is the extract through supercritical fluid extraction, has effect for reducing blood fat.This extract contains piperine, and multiple compositions such as piperine methyl ester include volatile oil, are semi-solid.This extract is to photo-labile by analysis, and volatile oil is volatile, in order to be made into solid preparation stable and controllable for quality, intends adopting beta-cyclodextrin inclusion compound technology to be made into clathrate.In order to select best clathrate process, this paper has investigated the optimum process condition that saturated water solution method, hand grinding method, ball mill legal system are equipped with Fructus Piperis Longi extract-Benexate Hydrochloride, now is discussed below:

1, saturated water solution method: get the distilled water that round-bottomed flask adds certain volume, be heated to more than 80 ℃, again predetermined amount of beta-cyclodextrin is dissolved in wherein, reduce to set point of temperature then, make into saturated solution.Get in the alcoholic solution that the 0.5g crude drug is dissolved in 3ml 95%, dropwise add then, drop in the above-mentioned saturated solution under stirring.After stirring certain hour, leave standstill 0～4 ℃ of cold preservation 24h after, filter, precipitate with a small amount of distilled water wash.The a small amount of ether washing of reuse clathrate places clathrate vacuum drying oven drying at room temperature 24h promptly to get the clathrate of off-white color then behind the natural drying.

2, hand grinding method: get the alcoholic solution that the 0.5g crude drug is dissolved in 3ml 95%.Get predetermined amount of beta-cyclodextrin in mortar, add a certain proportion of distilled water, grind to form pasty state, under grinding, dropwise add the alcoholic solution of crude drug then.After grinding certain hour, filter, and precipitate with a small amount of distilled water wash.The a small amount of ether washing of reuse clathrate behind the natural drying, then clathrate being placed with the phosphorus pentoxide is the vacuum drying oven drying at room temperature 24h of desiccant, promptly gets the clathrate of off-white color.

3, ball mill method: get predetermined amount of beta-cyclodextrin and add in the ball mill, add a certain proportion of distilled water again.Get in the alcoholic solution that the 0.5g crude drug is dissolved in 3ml 95%, add in the ball mill then.Set milling time, start ball mill.Grind and finish the back taking-up, operate same hand grinding method thereafter.

3 kinds of methods are carried out 3 times repeated experiments under best clathrate process separately, its enclose effect is compared.Confirm that through repeated trials each best clathrate process stablizes feasiblely, inclusion rate and productive rate can reach more than 70% and 75% respectively.The inclusion rate of saturated water solution method is the highest, is 79.21%, is about 75.23% but productive rate is relatively low; The hand grinding method productive rate is the highest, is 86.7%, and inclusion rate is 78.66%, and the inclusion rate of its inclusion rate and saturated water solution method is close; Ball mill method inclusion rate relatively low (inclusion rate is 74.68%), productive rate is 82.23%.

In the best clathrate process of optimization, beta-schardinger dextrin-is consistent with the optimal proportion of medicine in 3 kinds of methods, and promptly the ratio of β-CD and Fructus Piperis Longi extract is 8: 1 (gg-1).This can be with character relevant explain of medicine with the rate of charge and the medicine of beta-schardinger dextrin-, and with a kind of medicine, character is identical, though so different its rate of charges of inclusion method identical.

The productive rate of clathrate is relevant with the consumption of water.The consumption of saturated water solution method water is hand grinding method and ball mill method water consumption 22 times, therefore the productive rate of saturated water solution method is starkly lower than back 2 kinds of methods, though through cold preservation, but be dissolved in the clathrate that wherein discarded still greater than the back both, therefore cause productive rate lower, the someone think can be by adding organic solvent method with clathrate soluble in water " extruding " as much as possible, and bibliographical information [4] is arranged abroad with its Recycling Mother Solution use, can reduce the consumption of water, improve productive rate.

The inclusion rate of 3 kinds of methods has all reached more than 70%, and productive rate has all reached more than 75%, all is inclusion method preferably.The inclusion rate of hand grinding method and productive rate are all higher, and be simple and easy to do, is suitable for laboratory screening technology, but this method can only be hand-manipulated, is not suitable for suitability for industrialized production; Ball mill method efficient height can be used for suitability for industrialized production, but need appointed condition; The inclusion rate of saturated water solution method is higher, though productive rate is lower, can improve productive rate by recycling mother solution in the industry.Therefore, laboratory and factory can select diverse ways for use according to self instrument and equipment condition and needs.

Embodiment 15: contain the preparation of the tablet of Fructus Piperis Longi extract

Prescription: per 1000 slice prescriptions are composed as follows:

Benexate Hydrochloride 131.6g (containing Fructus Piperis Longi extract 20g)

Cross-linked pvp 40g

Lactose 20g

Microcrystalline Cellulose 80g

Magnesium stearate 1.4g

In right amount

25% alcoholic solution of 1% HPMC

Carry out pilot scale by selected prescription, technology, get the Benexate Hydrochloride 1316g of Fructus Piperis Longi extract; Cross-linked pvp 400g; Lactose 200g; Microcrystalline Cellulose 800g crosses 100 mesh sieves respectively, accurately crosses 80 mesh sieves after the weighing and mixes three times; HPMC with 1% (w/v, the preparation of 25% ethanol) system soft material, 16 mesh sieves are granulated; at 50 ℃ of baking oven inner dryings; granulate (control pellet moisture content is about 3%) is crossed 80 mesh sieves with magnesium stearate 14g, behind the granule mixing; tabletting; theoretical dosage is 10000, and finished product is 9590, bag water solublity white film clothing.The coating material Opadry is purchased in Shanghai Colorcon Coating Technology Co., Ltd (the product code name is 85G68918).

Embodiment 16: Fructus Piperis Longi extract is to the influence of hyperlipemia rat blood fat

In order to observe Fructus Piperis Longi extract effect for reducing blood fat of the present invention, 50 rats are divided into 5 groups at random: matched group, high lipid food group (model group), Fructus Piperis Longi extract low dose (20mgkg ^-1D ^-1), middle dosage (60mgkg ^-1D ^-1), heavy dose of (120mgkg ^-1D ^-1)+high lipid food group is measured the influence of Fructus Piperis Longi extract to serum triglycerides (TG), T-CHOL (TC), high density lipoprotein (HDL) and low density lipoprotein, LDL (LDL).The result shows that the extract of embodiment 1-13 has tangible blood fat reducing effect.The Fructus Piperis Longi extract that obtains with embodiment 5 is an example, and by table 1 as seen, model group is compared with matched group, and TC and TG content significantly raise, and the modeling success is described; And respectively organize TC, TG content and the model group that give Fructus Piperis Longi extract relatively have significance to reduce, and are respectively P＜0.01 and P＜0.05; The content of HDL-C and model group be P＜0.05 relatively, and increases with the content of Fructus Piperis Longi extract, has dose dependent; The content of LDL-C and model group be P＜0.05 and P＜0.01 relatively, and along with the content of Fructus Piperis Longi extract raises and reduces, illustrate that Fructus Piperis Longi extract has significant blood fat reducing effect, its mechanism may be metabolism and the conversion that has promoted between the lipoprotein on the one hand, may be in addition because suppressed intestinal cholesterol absorption, reduced the decomposition of having synthesized, promoted serum and liver tg of liver cholesterol.

Table 1 Fructus Piperis Longi extract to the influence of tentative high blood lipid model rat fat level (x ± s, n=10)

Annotate: compare with model group ^aP＜0.05, ^bP＜0.01.

Embodiment 17: Fructus Piperis Longi extract analgesia and antiinflammatory action

In order to observe Fructus Piperis Longi extract analgesia of the present invention and antiinflammatory action, adopt mice acetic acid to stimulate the analgesic activity of writhing method (table 2 and table 3) and mice hot plate stimulus method (table 4) research Fructus Piperis Longi extract; Its antiinflammatory action is studied in the inhibition of adopting the Fructus Piperis Longi extract Dichlorodiphenyl Acetate to cause mouse peritoneal capillary permeability (table 5 and table 6) and mice caused by dimethylbenzene xylene auricle edema (table 7).The result shows that the extract of embodiment 1-13 has tangible analgesia and antiinflammatory action.The Fructus Piperis Longi extract that obtains with embodiment 6 is an example, by table 2-7 as can be known: compare 100mgkg with negative control group ^-1～200mgkg ^-1Fructus Piperis Longi extract just can obviously prolong the pain thresholding (P＜0.05 or P＜0.01) of mice; 100mgkg ^-1Fructus Piperis Longi extract just can obviously suppress mouse peritoneal capillary permeability (P＜0.05); 200mgkg ^-1Fructus Piperis Longi extract just can obviously suppress mice caused by dimethylbenzene xylene auricle edema (P＜0.05).Conclusion: Fructus Piperis Longi extract has significantly analgesia and antiinflammatory action.

Table 2 Fructus Piperis Longi extract Dichlorodiphenyl Acetate induced mice is turned round the influence of body

Annotate: compare with the negative control group group ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001.

The turn round body of table 3 BBP different dosing time to the HAC induced mice influences

Annotate: compare with the negative control group group ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001.

Table 4 Fructus Piperis Longi extract causes the influence of the pain mice threshold of pain to hot plate

Annotate: compare with the negative control group group ^*P＜0.05, ^*P＜0.01.

The hyperfunction influence of table 5 Fructus Piperis Longi extract Dichlorodiphenyl Acetate induced mice abdominal cavity vascular permeability

The influence of table 6 Fructus Piperis Longi extract Dichlorodiphenyl Acetate induced mice abdominal cavity permeability

Table 7 Fructus Piperis Longi extract xylol causes the influence of mice ear

Embodiment 18: Fructus Piperis Longi extract is to the protective effect of gastric ulcer animal model

In order to observe the protective effect of Fructus Piperis Longi extract of the present invention, adopt the effect of answering acute gastric ulcer (table 8), ethanol gastric ulcer (table 9), reserpine gastric ulcer (table 9) and 4 kinds of scale-model investigations of rat pylorus ligation gastric ulcer (table 10) and estimating the Fructus Piperis Longi extract anti-gastric-ulcer to the gastric ulcer animal model.By traditional method modeling, administration, record ulcer is counted and area, and administration group and model group compare.The result shows that the extract of embodiment 1-13 has the effect of tangible control gastric ulcer.The Fructus Piperis Longi extract that obtains with embodiment 9 is an example, by table 8-10 as can be known: with model group relatively, Fructus Piperis Longi extract can suppress significantly and reduce that the ulcer of mice stress ulcer is counted, ulcer area, hemorrhagic focus size and extent of hemorrhage (P＜0.05 or P＜0.01) (seeing Table 8); With model group relatively, Fructus Piperis Longi extract can suppress significantly and reduce that the ulcer of mice ethanol gastric ulcer is counted, ulcer area and ulcer index (P＜0.05 or P＜0.01) (seeing Table 9); With model group relatively, Fructus Piperis Longi extract can suppress significantly and reduce that the ulcer of mice reserpine gastric ulcer is counted, ulcer area and ulcer index (P＜0.05 or P＜0.01) (seeing Table 9); With model group relatively, Fructus Piperis Longi extract can suppress significantly and reduce that the ulcer of rat pylorus ligation ulcer is counted, ulcer area and ulcer index (P＜0.05 or P＜0.01) (seeing Table 10).Conclusion: Fructus Piperis Longi extract has the effect of tangible control gastric ulcer.

Table 8 Fructus Piperis Longi extract to the influence of mouse experiment ulcer blood kitchen range (x ± s, n=10)

Compare with model group: ^*P＜0.05, ^*P＜0.01

Table 9 Fructus Piperis Longi extract to the exponential influence of mouse experiment ulcer (x ± s, n=10)

Compare with model group: ^*P＜0.05, ^*P＜0.01

Table 10 Fructus Piperis Longi extract to the influence of rat pylorus ligation gastric ulcer (x ± s, n=10)

Compare with model group: ^*P＜0.05, ^*P＜0.01

Embodiment 19: toxicity research

Rat acute toxicity test and long term toxicity test result of study all show the Fructus Piperis Longi extract safety range broad that embodiment 1-13 prepares.And long term toxicity test shows, rat successive administration 3 months, routine blood test, hepatic and renal function and heart, thymus, lungs, breastbone, pancreas, liver, spleen, kidney, adrenal gland, ovary, uterus, mammary gland, brain, hypophysis, optic nerve, stomach, large intestine, small intestinal, bladder, thyroid, bone formation microscopy, no abnormality seen.Extracting the Fructus Piperis Longi extract that obtains with embodiment 8 is example, and the safe dose of rat long term toxicity test is 300mgkg ^-1D ^-1

Claims (21)

1. a Fructus Piperis Longi extract is characterized in that it prepares by following method, and this method comprises the following steps:

Step (a) is 70%～100% ethanol extraction with Fructus Piperis Longi concentration, to obtain ethanol extract;

Step (b) is with the ethanol extract decompression recycling ethanol, to obtain fluid extract;

Step (c) was separated out crystallization in 24～96 hours with the fluid extract placement;

Step (d) will be separated out fluid extract after the crystallization with 10～20%KOH aqueous solution saponification of 4～10 times of weight, to obtain the Fructus Piperis Longi non-saponifiable matter;

Step (e) with Fructus Piperis Longi non-saponifiable matter condition is: pressure=9～32MPa, and flow velocity=2～6mL/min, Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL=1: 1～1: 4, temperature is 35～77 ℃ CO ₂The supercritical rectification method carries out separation and purification, the Fructus Piperis Longi extract that obtains.

2. the described Fructus Piperis Longi extract of claim 1, it is characterized in that, this extract mainly contains methyl piperate and seven kinds of piperlongumine chemical compounds: Piperlonguminine, piperine, the decadinene amide, the peaceful alkali of Fructus Piperis promise, the quinine element, 18 carbon triolefin amide and eight chemical compounds of 20 carbon triolefin amide, by weight percentage, eight kinds of chemical compound gross weights account for 29.07～87.80% of Fructus Piperis Longi extract gross weight, wherein Piperlonguminine accounts for 0.95～3.43% of Fructus Piperis Longi extract gross weight, piperine accounts for 14.52～42.78% of Fructus Piperis Longi extract gross weight, and methyl piperate accounts for 0.82～3.11% of Fructus Piperis Longi extract gross weight, and the decadinene amide accounts for 1.36～4.27% of Fructus Piperis Longi extract gross weight, the peaceful alkali of Fructus Piperis promise accounts for 2.96～8.03% of Fructus Piperis Longi extract gross weight, 3.17～10.94%, the 20 carbon triolefin amide that 1.04～3.26%, the 18 carbon triolefin amide that the quinine element accounts for the Fructus Piperis Longi extract gross weight account for the Fructus Piperis Longi extract gross weight account for 4.25～12.42% of Fructus Piperis Longi extract gross weight.

3. claim 1 or 2 described Fructus Piperis Longi extracts, wherein eight kinds of chemical compound gross weights account for 49.64～71.89% of Fructus Piperis Longi extract gross weight.

4. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.88～2.54% to Piperlonguminine.

5. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 24.16～35.16% to piperine.

6. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.72～2.35% to methyl piperate.

7. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 2.21～2.67% to the decadinene amide.

8. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 4.58～6.75% to the peaceful alkali of Fructus Piperis promise.

9. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 1.83～2.44% to the quinine element.

10. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 6.05～9.43% to 18 carbon triolefin amide.

11. claim 1 or 2 described Fructus Piperis Longi extracts, wherein to account for the percentage by weight of Fructus Piperis Longi extract gross weight be 7.21～10.55% to 20 carbon triolefin amide.

12. by each described Fructus Piperis Longi extract of claim 1-11 make be used for the treatment of hyperlipidemia, analgesia, antiinflammatory or gastric ulcer pharmaceutical composition.

13. the described pharmaceutical composition of claim 12 is characterized in that said composition is the pharmaceutical preparation of oral administration.

14. the described pharmaceutical composition of claim 13, wherein the pharmaceutical preparation of oral administration is selected from a kind of in tablet and the powder.

15. a method for preparing each described Fructus Piperis Longi extract of claim 1-11 is characterized in that this method comprises the following steps:

Step (a) is 70%～100% ethanol extraction with Fructus Piperis Longi concentration, to obtain ethanol extract;

Step (b) is with the ethanol extract decompression recycling ethanol, to obtain fluid extract;

Step (c) is placed fluid extract and was separated out crystallization in 24～96 hours;

Step (d) will be separated out fluid extract after the crystallization with 10～20%KOH aqueous solution saponification of 4～10 times of weight, to obtain the Fructus Piperis Longi non-saponifiable matter;

Step (e) with Fructus Piperis Longi non-saponifiable matter condition is: pressure=9～32MPa, flow velocity=2～6mL/min, Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL=1: 1～1: 4, temperature is that 35～77 ℃ CO2 supercritical rectification method carries out separation and purification, obtains each described Fructus Piperis Longi extract of claim 1-10.

16. the described method for preparing Fructus Piperis Longi extract of claim 15 is characterized in that used concentration of alcohol is 95% in the step (a).

17. the described method for preparing Fructus Piperis Longi extract of claim 15 is characterized in that it is 48 hours that the crystalline time is separated out in the middle placement of step (c).

18. the described method for preparing Fructus Piperis Longi extract of claim 15 is characterized in that used KOH concentration is 15% in the step (d), the KOH consumption is 6 times.

19. the described method for preparing Fructus Piperis Longi extract of claim 15, it is characterized in that CO2 supercritical rectification method process conditions are in the step (e): pressure=11.5MPa, flow velocity=3mL/min, Fructus Piperis Longi non-saponifiable matter extractum quality g: ethanol volume mL=1: 2, thermograde=35-49-63-77 ℃.

20. the application of each described Fructus Piperis Longi extract of claim 1-11 in the preparation medicine, wherein said medicine is used for the treatment of hyperlipidemia, analgesia, antiinflammatory or gastric ulcer.

21. the application of each described pharmaceutical composition of claim 12-14 in the preparation medicine, wherein said medicine is used for the treatment of hyperlipidemia, analgesia, antiinflammatory or gastric ulcer.

Images