CN102716272B - Traditional Chinese medicine composition for senescence delaying and antifatigue as well as soft capsule and preparation method of traditional Chinese medicine composition - Google Patents

Abstract

The invention relates to a traditional Chinese medicine composition for senescence delaying and antifatigue as well as a soft capsule and a preparation method of the traditional Chinese medicine composition and belongs to the technical field of traditional Chinese medicine. The traditional Chinese medicine composition consists of the following raw material medicine in parts by weight: 2 parts of ginseng, 10 parts of grape seed, 1 parts of caltrop and 1 parts of herba epimedii, wherein the soft capsule comprises pharmaceutically acceptable carriers and active ingredients prepared from the ginseng, the grape seed, the caltrop and the herba epimedii. The traditional Chinese medicine composition and the soft capsule of the traditional Chinese medicine composition have the advantage that the effects of senescence delaying and antifatigue are realized.

Description

The Chinese medicine composition of a kind of slow down aging and resisting fatigue, its soft capsule and preparation method

Technical field

The invention belongs to technical field of Chinese medicine, be specifically related to the Chinese medicine composition of a kind of slow down aging and resisting fatigue, its soft capsule and preparation method.

Background technology

Old and feeble, also claim aging (Aging), refer to that the mankind are after reaching maturity, along with the increase at age or the growth body of time there is changes in microstructure, hypofunction, adaptive capacity weaken, organ stress ability declines, viability is lost gradually, are easier to dead process.Since "Nei Jing", practical experience discloses already, from childhood to the young middle age, along with filling gradually of the five internal organs is contained and body also crescendo Sheng.Body aging is the result that the five internal organs void declines, and each tissue and function of visceral controllings, all without unattenuated.It is very complicated that modern medicine study also shows to affect old and feeble factor.Medical Aging Mechanism and the research of Chinese medicine slow down aging thereof are thought, people is an organic whole, and the five internal organs are then the hosting centers of this organic whole.The old and feeble one as complicated biosis change, is the result of multifactor functioning, the summation of each internal organs aging that to be the aging of individuality be.Find from the integral level of organ and the research of molecular cell, old and feeble and gene, free radical, endocrine function are lacked of proper care, and the factors such as immunologic function disorder are relevant.Research finds, a large amount of oxide radical that human body metabolism produces, and destroys cell function, causes the heart organ of human body aging.Endocrine function is lacked of proper care, and standing in the breach affected is also the deterioration of human body internal organs, along with the loss of health accumulates, causes aging.

Fatigue is the formidable enemy of 21 century human health.Fatigue not still medical problem Ye Shi social problem, not only make the quality of life of people reduce, also constrain expanding economy, and then affect the process of social development, preventing and treating fatigue becomes the task of top priority.Chronic fatigue syndrome (Chronic Fatigue Syndrome, CFS) be a kind of with agnogenic, continue or recurrent exerbation at least 6 months and have a rest can not the obvious fatigue alleviating disease that is principal character, it is often with short-term memory loss, to focus one's attention on the cognitive dysfunction such as difficulty, the sleep of not recovering from fatigue, the influenza-like symptom of pharyngalgia, lymph enlargement, or the Somatic discomfort symptom such as headache, muscle arthralgia.In modern society, along with dog-eat-dog, the increasing of pressure, the quickening of rhythm, mental, muscle power is in anxiety for a long time, the people of fatigue state just increases with day, and usually because tired nothing is in a helpless situation hopeless because looking into.Tired treatment basic principle should be dredge to mend to combine, and tonifying deficiency is with thin regulating QI machine.Reason internal organs, tonifying deficiency, Pingyin is damaged, and recovers healthy object.

Summary of the invention

The sub-health status such as fatigue, aging that the present invention causes due to the problem such as operating pressure and social pressure to solve current social, provides the Chinese medicine composition of a kind of slow down aging and resisting fatigue.

Second object of the present invention is the Chinese medicinal soft capsule disclosing a kind of slow down aging and resisting fatigue.

Second object of the present invention is the preparation method of the Chinese medicinal soft capsule disclosing a kind of slow down aging and resisting fatigue.

In order to realize the object of foregoing invention, the present invention takes following technical scheme:

A Chinese medicine composition for slow down aging and resisting fatigue, described Chinese medicine composition is made up of the crude drug of following weight portion: Radix Ginseng 2 parts, Semen Vitis viniferae 10 parts of g, Fructus Tribuli 1 part of g and Herba Epimedii 1 part.

A Chinese medicinal soft capsule for slow down aging and resisting fatigue, described Chinese medicinal soft capsule comprises active component and pharmaceutically acceptable carrier, and wherein, described active component is prepared by following method by Radix Ginseng, Semen Vitis viniferae, Fructus Tribuli and Herba Epimedii:

(1), insert in extraction pot by Radix Ginseng decoction pieces, with 75% ethanol extraction twice, add ethanol 6 times amount at every turn, extract 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and when decompression recycling ethanol to 60 is spent, the concentrated solution of relative density 1.15 is for subsequent use; Residue stays and does water extraction use;

(2) after, getting the middle Radix Ginseng residue mixing of Semen Vitis viniferae, Fructus Tribuli coarse grain, Herba Epimedii crin and step (1), extracting in water 3 times, add water 6 times amount at every turn, carry 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and leave standstill 4 hours, Aspirate supernatant, the extraction concentrated solution of relative density 1.15 when evaporation and concentration to 60 is spent; This extraction concentrated solution is added the ethanol of 95%, make alcohol content be 60%, leave standstill 24 hours, Aspirate supernatant postprecipitation filters, and filtrate and supernatant merge, decompression recycling ethanol, to the concentrated solution of relative density 1.15 when 60 degree;

(3) after the concentrated solution that the concentrated solution, by step (2) obtained and step (1) obtain mixes, the spray-dried dry extract obtaining active component.

Chinese medicinal soft capsule described in technique scheme, wherein, containing Total saponin Rb in every 100g active component ₁430mg, procyanidin 253mg.

Chinese medicinal soft capsule described in technique scheme, wherein, described pharmaceutically acceptable carrier is made up of soybean oil, gelatin, glycerol and distilled water.

The preparation method of the Chinese medicinal soft capsule of a kind of slow down aging and resisting fatigue, comprise the step being obtained Chinese medicinal soft capsule agent by active component and pharmaceutically acceptable carrier, wherein, described preparation method also comprises the preparation process of active component, and wherein the preparation process of active component comprises:

(1), insert in extraction pot by Radix Ginseng decoction pieces, with 75% ethanol extraction twice, add ethanol 6 times amount at every turn, extract 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and when decompression recycling ethanol to 60 is spent, the concentrated solution of relative density 1.15 is for subsequent use; Residue stays and does water extraction use;

(2) after, getting the middle Radix Ginseng residue mixing of Semen Vitis viniferae, Fructus Tribuli coarse grain, Herba Epimedii crin and step (1), extracting in water 3 times, add water 6 times amount at every turn, carry 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and leave standstill 4 hours, Aspirate supernatant, the extraction concentrated solution of relative density 1.15 when evaporation and concentration to 60 is spent; This extraction concentrated solution is added the ethanol of 95%, make alcohol content be 60%, leave standstill 24 hours, Aspirate supernatant postprecipitation filters, and filtrate and supernatant merge, decompression recycling ethanol, to the concentrated solution of relative density 1.15 when 60 degree;

(3) after the concentrated solution that the concentrated solution, by step (2) obtained and step (1) obtain mixes, the spray-dried dry extract obtaining active component.

Preparation method described in technique scheme, wherein, the step being obtained Chinese medicinal soft capsule by active component and pharmaceutically acceptable carrier comprises:

(a), get the active component dry extract preparing gained and mix with the soybean oil of pharmaceutically acceptable amount, through colloid mill, grinding evenly, obtains the suspensoid of dry extract oil;

(b), according to gelatin: glycerol: distilled water=1: the weight ratio of 0.6: 1, takes each raw material, is first soaked in water by gelatin 12 hours, after making it fully expand, the glycerol of additional proportion amount, adds distilled water, is heated to 70-80 degree, stirring, melting, mixing, insulation 1-2 hour, leaves standstill, removing floating foam, filter, obtain glue shell feed liquid heat preservation for standby use;

C glue shell feed liquid that (), the suspensoid of dry extract oil step (a) obtained and step (b) obtain is placed in automatic rotary respectively and presses capsule Bigpian, room temperature 23 degree, under the condition of relative humidity 40%, be pressed into soft gelatin capsule, obtain Chinese medicinal soft capsule.

The principle that Chinese medicine composition of the present invention, soft capsule have slow down aging and anti-fatigue health-caring effect is that in raw material, each crude drug has the following property of medicine:

1, Radix Ginseng is that the drying of araliaceae ginseng plant's Radix Ginseng is followed, it is traditional rare Chinese medicine, its sweet in the mouth, micro-hardship, warm in nature, have strongly invigorating primordial QI, admittedly take off effects such as promoting the production of body fluid, this product mainly contains multiple soaping agents, in addition, still containing the volatile ingredient such as Panaxynol (Panaxynol), beta-elemene (Elem-ene), and monosaccharide and disaccharide, three polysaccharide, low molecular peptide, several amino acids; Also have the compositions such as palmitic acid, stearic acid, glycoside, its effective ingredient is ginsenoside.Ginsenoside is one of principle active component of Radix Ginseng, and the major physiological that it has Radix Ginseng is active, and modern pharmacological research proves; Ginsenoside's energy extending life life time, obviously suppresses liver, brain foundation of microsomal Lipid Peroxidation, and can improve the content of SOD in body.Radix Ginseng begins to be loaded in the first monograph on materia medica Shennong's Herbal of China, it is the famous medication of prolonging life of Ancient Times in China, along with the development of modern science and technology, the particularly application of Protocols in Molecular Biology, Radix Ginseng research is made to be deep into cell, molecule, gene level, since the seventies, report discloses, ginsenoside adds in culture medium can make diploid cell life, in the culture medium of the Radix Ginseng extract of debita spissitudo, diploid cell mitogen activation and the speed of growth strengthen, human embryonic lung fibroblast density is significantly higher than matched group, result shows that appropriate ginsenoside can extend cell survival, the important component part Rb1 of Ginsengdiol histsaponin, not only there is T suppression cell lipid peroxidation, the ability of scavenging free radicals, catalase can also be made, activity of glutathione peroxidase increases.Ginseng aqueous extract is by strengthening hormonal system, as hypothalamic pituitary gonadal axis, thyroid system and adrenocortical function regulate the multiple metabolic activity of body, thus reach the object of anti-ageing life-prolonging, ginseng aqueous extract can also promote body's immunity, strengthen resistance against diseases, it all has stimulation to humoral immunization and cellular immunization, can strengthening vital QI to eliminate pathogenic factors, delay cell degradation, prevent cell from declining extremely.The Macro and microelements that Radix Ginseng contains also is one of antidotal major reason, by trace element supplement, can regulate metabolic activity relevant with it further, thus reach the object of anti-ageing life-prolonging.

Ginseng qi-tonifying hemopoietic, help intensive culture god, bodily fuctions and the effect of " adaptogen " sample can be strengthened, so-called " adaptogen " sample effect, be exactly can enhancing body to the defence capability of various harmful factor, improve organism adaptation, promote the recovery of normal function, and the cerebral cortex of dysfunction can be adjusted, improve the function of brain, intelligence of old people and hypomnesis and bradykinesia are had some improvement, per os is to mice Radix Ginseng extractive solution (containing ginsenoside 0.3-1.5mg/kg.bw.d) gavage 30d continuously, mice walk time can be continued, the generation of blood lactase acid when reducing travelling, show that ginsenoside has obvious antifatigue effect.

2, Semen Vitis viniferae extract (procyanidin OPC, proanthocyanidin) is a kind of new and effective antioxidant, has extremely strong activity in vivo and free radical scavenging effect.Research confirms, OPCkeyi reduces the danger of heart disease, cancer, arthritis and degenerative disease.OPC also has enhancing blood vessel wall resistance, reduces capillary fragility, keeps the suitable permeability of blood capillary and strengthen skin elasticity and effect that is smoothness and immunity of organisms; Widely, its disease-prevention health effect basis is exactly the ability knowing free radical in the application of Grape Seed Extract Procyanidin, and effect of its slow down aging realizes by removing interior free yl, has good delaying senility function; In addition, it also has good bioavailability, is easily combined with collagen protein, stabilizing cell membrane and antienzyme activity.In addition OPC is owing to having stronger antioxidation; be conducive to the removing of active oxygen; body lipoid peroxidization resistant is strengthened; reduce the generation machine of free radical during motion to the damaging action of tissue; the generation of delay fatigue effectively; experiment shows the procyanidin energy significant prolongation mice burden swimming time; reduce tired Mus Serum lactic acid content and increase hepatocyte glycogen content; thus prove that procyanidin has antifatigue effect, its mechanism of action may mainly by scavenging free radicals and free radical is caused organize the protective effect of peroxide injury to realize.

3, Fructus Tribuli (Tribulusterrestrisl) is also known as Fructus Atriplicis Sibiricae, Fructus Tribuli, it is zygophyllaceae (zygophyllaceae) Tribulus plant, tool suppressing the hyperactive liver resolving depression, promoting blood circulation by removing wind, expelling phlegm for arresting cough, improving eyesight, diuresis, antipruritic effect, to dispel renal cold, town is conspicuous complies with, and strong, diuresis is relieved oedema or abdominal distension through diuresis or purgation, for edema, water urine, urine retention, the lumbago that renal cold causes, seminal emission, sexual impotence.The active chemical of Fructus Tribuli mainly contains alkaloid, flavonoid, saponins etc., then prevents and treats the aspects such as cardiovascular and cerebrovascular disease, defying age and therapeutic hypofunction and all demonstrate good efficacy, and the research of Fructus Tribuli and effective ingredient thereof is main mainly with steroidal saponin.

4, Herba Epimedii is the traditional tonic medicine of Chinese tradition, and history tree is all on the books, begins to be loaded in Shennong's Herbal, another name Herba Epimedii, for starving Berberidaceae (Berberidaceae) Epimedium (Epimedium L.) plant.Compendium of Material Medica is recorded: stem, leaf are used as medicine, and Xin Wen is nontoxic, and have hard muscles and bones, beneficial vital essence, mend the effects such as waist knee joint, heart tonifying power, husband for a long time clothes makes us loss of fecundity, the exhausted positive loss of fecundity of husband, and old dusk octogenarian, the middle age is forgetful, and extremity are meciless, and hemiplegia is unsuccessful, and insufficiency of kidney-YANG person should it.Modern pharmacological research shows: raw product, without promoting and inhibition function, namely cold in nature; Processed product, after warm moral Adeps caprae seu ovis is concocted, promotes sexual function, namely warm in nature.Domestic and international research shows, Main Chemical Constituents of Herb Epimedii is flavone compound, lignin and alkaloid; Pharmacological research in recent years about Herba Epimedii shows, Herba Epimedii has multiple pharmacodynamic feature, mainly concentrates on immunity, reproductive system, nucleic acid metabolism, cardio-cerebrovascular and anti-ageing aspect of waiting for a long time.Epimedium flavone significantly can recover the function of D-gal Aging model mice T and B lymphocyte proliferation reaction, improve the activity of the total SOD of liver simultaneously, the lipofuscin reducing the tissue such as the heart, liver is formed, epimedium brevicornum polysaccharide liposome (EPSS) significantly improves older animals erythrocyte and SOD activities of liver, improve the activity of aged mouse GSH-Px, obviously can reduce the content of lipid peroxide (LPO) in older animals serum and hepatic tissue, reduce the content of aged mouse cardiac muscle lipofuscin (LF), have the effect of antioxidation, slow down aging, resisting fatigue.

In sum, containing the functional component such as ginsenoside, procyanidin in the present invention, through modern medicine study, there is slow down aging, the effect that resisting fatigue, promotion body function recover.

Chinese medicinal soft capsule specification of the present invention is 0.7g/ grain; Have effect of slow down aging, resisting fatigue, suitable population is middle-aged and elderly people, fatiguability person; The crowd that is not suitable for is children, anemia of pregnant woman; Every day 2 times, each 1, (medicine) being taken before meal is with better.

The present invention has following beneficial effect:

1, have can slow down aging and resisting fatigue for the soft capsule of Chinese medicine composition of the present invention and extract thereof, has the effect promoting that bodily fuctions recovers.

Accompanying drawing illustrates:

1, Fig. 1 is the production technological process of soft capsule of the present invention;

2, Fig. 2 is the HPLC chromatogram in ginsenoside Rb1 in soft capsule content of the present invention (Total saponin Rb1) assay.

Detailed description of the invention:

Be convenient to for making technical scheme of the present invention understand, below in conjunction with specific embodiment, the present invention is further illustrated.

embodiment 1:the preparation of the Chinese medicine composition of slow down aging and resisting fatigue:

Weighting raw materials Radix Ginseng 2 parts, Semen Vitis viniferae 10 parts, Fructus Tribuli 1 part and Herba Epimedii 1 part.

embodiment 2:the preparation of the Chinese medicinal soft capsule of slow down aging and resisting fatigue:

A preparation method for the Chinese medicinal soft capsule of slow down aging and resisting fatigue, comprises the steps

(1), insert in extraction pot by Radix Ginseng decoction pieces, with 75% ethanol extraction twice, add ethanol 6 times amount at every turn, extract 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and when decompression recycling ethanol to 60 is spent, the concentrated solution of relative density 1.15 is for subsequent use; Residue stays and does water extraction use;

(2) after, getting the middle Radix Ginseng residue mixing of Semen Vitis viniferae, Fructus Tribuli coarse grain, Herba Epimedii crin and step (1), extracting in water 3 times, add water 6 times amount at every turn, carry 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and leave standstill 4 hours, Aspirate supernatant, relative density 1.15 when evaporation and concentration to 60 is spent; This extraction concentrated solution is added the ethanol of 95%, make alcohol content be 60%, leave standstill 24 hours, Aspirate supernatant postprecipitation filters, and filtrate and supernatant merge, decompression recycling ethanol, to the concentrated solution of relative density 1.15 when 60 degree;

(3) after the concentrated solution that the concentrated solution, by step (2) obtained and step (1) obtain mixes, the spray-dried dry extract obtaining active component of concentrated solution;

(4), get the active component dry extract preparing gained and mix with the soybean oil of pharmaceutically acceptable amount, through colloid mill, grinding evenly, obtains the suspensoid of dry extract oil;

(5), according to gelatin: glycerol: distilled water=1: the weight ratio of 0.6: 1, take each raw material, first gelatin is soaked in water 12 hours, after making it fully expand, the glycerol of additional proportion amount, adds distilled water, is heated to 70-80 degree, stirring, melting, mixing, insulation 1-2 hour, leaves standstill, removing floating foam, filter, obtain glue shell feed liquid heat preservation for standby use;

(6) the glue shell feed liquid that the suspensoid of the dry extract oil, step (4) obtained and step (5) obtain is placed in automatic rotary respectively and presses capsule Bigpian, room temperature 23 degree, under the condition of relative humidity 40%, be pressed into soft gelatin capsule, obtain Chinese medicinal soft capsule.

In Chinese medicinal soft capsule of the present invention, every 100g active component contains Total saponin 430mg, and the content of procyanidin 253mg carries out mensuration gained by the following method:

One, the mensuration of ginsenoside:

1, the scope of application:

This method provides the assay method that ginseng lozenge, ginseng infusion, Radix Ginseng tea, Panax Capsule etc. take Radix Ginseng as the HPLC of the content of ginsenoside in the health food of primary raw material.

This method is applicable to the assay method that ginseng lozenge, ginseng infusion, Radix Ginseng tea, Panax Capsule etc. take Radix Ginseng as the HPLC of the content of ginsenoside in the health food of primary raw material.

The limit of identification of six kinds of Saponins of this method is: 10mg/kg.

The optimum linear scope of six kinds of Saponins of this method: 0.1mg/mL ~ 1mg/mL.

2, principle:

Ginsenoside in sample is dissolved, extracts, after purified treatment, use gradient elution reversed-phase high-performance liquid chromatography to be separated, UV-detector (UV) detects, qualitative according to the retention time of chromatographic peak, outer marking quantitative, is applicable to Ginsenoside Re, R g1 in health food, Rb ₁, Rc, Rb2, Rd Simultaneous Quantitative Analysis.

3, reagent:

Test water is deionized water.

3.1, acetonitrile: chromatographically pure, 200nm absorbance is 0.021.

3.2, methanol: analytical pure.

3.3,101 macroporous adsorbent resins.

3.4, mobile phase of high performance liquid chromatography: gradient elution A liquid is acetonitrile, B liquid is water.

3.5, Ginsenoside Re, R g1, Rb1, Rc, Rb2, Rd standard substance: content is greater than 98% (HPLC).

3.6, the preparation of Ginsenoside Re, R g1, Rb1, Rc, Rb2, Rd standard solution:

Preparation Ginsenoside Re, R g1, Rb1, Rc, Rb2, Rd standard reserving solution, concentration is respectively 10mg/mL; Be mixed with hybrid standard serial solution with this storing solution again, concentration range is 0.1mg/mL ~ 1mg/mL; All standard solution are all prepared with methanol.

4, instrument and equipment

4.1, high performance liquid chromatograph: two high pressure pump, attached UV-detector.

4.2, ultrasonic cleaner.

4.3, centrifuge.

4.4, water-bath.

5, analytical procedure

Tablet or capsule 's content pulverize are got in 5.1 solid sample process, and cross 20 mesh sieves, accurately take this powder-like in right amount in 50mL tool plug test tube, add water 50mL supersound extraction 30 minutes in ultrasonic cleaner, take out, recover after room temperature until solution, accurate taking-up 10mL, by D-101 macroporous adsorbent resin decontaminating column, (macroporous adsorbent resin first soaks through methanol before using, washing, dress up the long pillar of 10cm), pillar first rinses with 10mL water, after discarding water liquid, with 70% methanol 25mL eluting Saponin, collect methanol solution, evaporate to dryness in water-bath, remain with dissolve with methanol and be settled to 5mL, the centrifugal rear mistake of this sample liquid 0.5 μm of film, filtrate carries out chromatography.

5.2, a certain amount of sample evaporate to dryness in water-bath is got in liquor sample process, and residual with 50mL water supersound extraction 30min, remaining steps is identical with 5.1.

5.3, measure

5.3.1, liquid chromatograph reference conditions

5.3.1.1, chromatographic column: anti-phase C ₁₈post, 4.6 × 250mm, 5 μm

5.3.1.2, UV-detector: determined wavelength 203nm

5.3.1.3 gradient conditions:

5.3.1.4, column temperature: 35 DEG C

5.3.2, chromatography

5.3.2.1, the preparation of standard curve:

Hybrid standard serial solution is all got 5 μ L and carry out HPLC analysis, with peak area, concentration is made to the standard regression curve of each Saponin.

5.3.2.2, Specimen Determination:

Get 5 μ L sample scavenging solutions and carry out efficient liquid phase chromatographic analysis, qualitative with absolute retention time, with the content of peak area by Ginsenoside Re, R g1, Rb1, Rc, Rb2, Rd in the standard curve quantitative Analysis sample of each Saponin.

6, analysis result statement:

6.1, calculate

In formula: the content of each ginsenoside in C-sample solution, mg/mL;

M-sample mass, g;

Content (the g/100g)=C of total ginsenoside in sample _re+ C _rg1+ C _rb1+ C _rc+ C _rb2+ C _rd

In formula: C _re(g/100g): the content of Re in sample;

C _rg1(g/100g): the content of Rg1 in sample;

C _rb1(g/100g): the content of Rb1 in sample;

C _rc(g/100g): the content of Rc in sample;

C _rb2(g/100g): the content of Rb2 in sample;

C _rd(g/100g): the content of Rd in sample.

6.2, result represents: result of calculation retains three position effective digitals.

7, chromatogram: as shown in Figure 2.

Through detecting, ginsenoside Rb1 (i.e. Total saponin Rb in product the sample of three batches (0.7g/ grain × 60 × 12 bottles × 3 batches) ₁) content as following table:

Ginsenoside Rb ₁(mg/kg)

Two, the assay method of procyanidin content:

1, scope:

This method provides the assay method of health food procyanidins

This method is applicable to the assay of health food procyanidins

This method limit of identification is 3 μ g, and concentration limit is 3 μ g/mL

This method optimum linear scope: 3-150 μ g/Ml

2, principle:

Procyanidin is the polymer containing catechin and epicatechin units.Procyanidin itself is colourless, but through with after hot acid treatment, can generate wine-colored anthocyanidin ion.The anthocyanidin ion that this method spectrophotometry procyanidin generates in hydrolytic process.Calculate sample procyanidins content.

3, reagent:

3.1, methanol analytical pure.

3.2, n-butyl alcohol analytical pure.

3.3, hydrochloric acid analytical pure.

3.4, ammonium ferric sulfate NH ₄fe (SO ₄) ₂12H ₂o solution: be the solution that 2mol/L hydrochloric acid is made into 2% (w/v) by concentration.

3.5, procyanidin standard substance Semen Vitis viniferae extract, purity 95%/

4, instrument:

4.1, spectrophotometer.

4.2, reflux.

5, analytical procedure:

5.1, the preparation of sample:

5.1.1, tablet gets 20 samples, grinds to form powdery

5.1.2, capsule extrudes 20 capsules contents, grinds or stirs, as content oil-containing, content should be extruded as far as possible

5.1.3, oral liquid shakes up rear sampling

5.2, extract:

5.2.1, powdery samples takes 50-100mg sample and is placed in 50mL volumetric flask, adds 30mL methanol, supersound process 20min, lets cool to room temperature, add methanol to scale, shake up, centrifugal or to get supernatant after being placed to clarification for subsequent use.

5.2.2, oil-containing sample takes 50mg sample and is placed in small beaker, divides and stirs for several times, washed by procyanidin in 50mL volumetric flask, until methanol extract liquid is colourless, adds methanol to scale, shake up with 20mL methanol.

5.2.3, oral liquid draws appropriate sample liquid (sampling amount is no more than 1mL) and is placed in 50mL volumetric flask, adds methanol to scale, shakes up.

5.3, measure:

5.3.1, standard curve takes procyanidin standard substance 10.0mg and is dissolved in 10mL methanol, draw this solution 0,0.1,0.25,0.5,1.0,1.5mL is placed in 10mL volumetric flask, add methanol to scale, shake up, respectively get 1mL to measure, identical with Specimen Determination method.

5.3.2, Specimen Determination by n-butyl alcohol and hydrochloric acid according to 95: 5 volume ratio mix after, take out 6mL and be placed in tool plug conical flask, add 0.2mL ammonium ferric sulfate solution and 1mL sample solution again, mixing, put boiling water bath backflow, after exact heat 40min, put in frozen water immediately and cool, after the complete 15min of heating, survey absorbance in 546nm wavelength place, calculated the content of sample procyanidins by standard curve, develop the color stable in 1 hour.

6, analysis result statement:

Sample procyanidins measurement result calculates by (1) formula:

$X(\%)=\frac{m_1\×v\×1000}{m\×1000\×1000}\×100...\left(1\right)$

In formula: the percentage composition of X-sample procyanidins, g/100g;

M ₁the amount of-reactant mixture procyanidins, μ g;

V-treats the cumulative volume of sample measuring liquid, mL;

The quality of m-sample, mg;

6.2, result represents:

Result of calculation retains three position effective digitals.

7, technical parameter

Relative standard deviation: < 10%.

The response rate: 84.6-94.4%.

Through detecting the sample of three batches (0.7g/ grain × 60 × 12 bottles × 3 batches), the content of product procyanidins is as following table:

Procyanidin (mg/kg)

In order to the beneficial effect that Chinese medicinal soft capsule of the present invention has better is described, below by way of concrete test example, the slow down aging that the present invention has and resisting fatigue effect are described:

Following test example all with " function of health food assessment process and the method for inspection " for test basis, inspection body is Tianjin Disease Prevention And Control Center (former Tianjin Center for Disease Control and Prevention).

the effect of the slow down aging of test example one, Chinese medicinal soft capsule of the present invention:

The inspection item of this test are the zoopery of Chinese medicinal soft capsule delaying senility function of the present invention.

1, material

1.1, sample: Chinese medicinal soft capsule prepared by the present invention, content is oily thick liquid.

1.2, experimental animal: secondary Male Kunming strain mice 75, body weight 18-22g, at about 3 monthly ages, is provided by Military Medical Science Institute's experimental animal center, the quality certification number: SCSK (army) 2002-001.Oregon wild type Drosophila melanogaster (Drosophila melanogaster), is provided by Shanghai Railway Univ's medical college.

1.3, laboratory animal room: SPF level laboratory animal room, the quality certification number: dynamic No. 015th, the word of doctor, temperature 20.8-22.5 DEG C, humidity 52-58%.Laboratory animal feedstuff is provided by Tianjin Huarong Animal Science company limited.

1.4, dosage choice:

1.4.1, drosophila survival experiment: Chinese medicinal soft capsule human body recommended intake of the present invention is 1.4g/d, if four concentration groups, is respectively 0.016%, 0.047%, 0.140% and 0.420%, separately establishes blank group.

1.4.2, peroxidating cosmetics (MDA) content and antioxidase (SOD) vitality test: Chinese medicinal soft capsule human body recommended intake of the present invention is 1.4g/60kg.BW.d, if three dosage groups, 0.117,0.233,0.70g/kg.BW be respectively:, be namely equivalent to 5,10,30 times of human intaking amount respectively.Tested material Oleum Arachidis hypogaeae semen is made into suspension, equal per os gavage, gavage amount 0.1ml/10g.BW.

1.5, key instrument reagent:

1.5.1, drosophila survival experiment: biochemical cultivation case (25 ± 1 DEG C, temperature 45-75%), cultivation vial (flat) 2.5 × 10cm, electronic balance, ether, propanoic acid.

1.5.2, antioxidation experiment: malonaldehyde (MDA) test kit, superoxide dismutase (SOD) test kit, by Tianjin, TBD Bioisystech Co., Ltd provides; UV-1201 ultraviolet-uisible spectrophotometer.

1.6, test method:

1.6.1, drosophila survival experiment: collect the new fruit bat adult sprouted wings in 8 hours, distinguish male and female random packet under etherization, each concentration group attacks each 200 of breast fruit bat, is divided in 10 cultivation vials, often pipe 20.Adult is weighed and gives tested material from point nest is after 2 weeks, fruit bat puts into tested material cultivation and namely rear experiment formally starts, matched group gives conventional corn powder culture medium, and test group gives the culture medium containing 0.016%, 0.047%, 0.140% and 0.420% tested material respectively.Experiment condition: temperature 25 ± 1 DEG C, relative humidity 54-66%.The death toll of timing every day observed and recorded fruit bat, changes a subculture, observes fruit bat all dead always for every 4 days.Experiment terminates, and calculates the dead natural law of half, average life and Mean longest life (often the Average Survival natural law of last 20 the dead fruit bats of group is the maximum life span of this group).

1.6.2, D-galactose modeling experiment: get secondary Kunming kind female mice 75, be divided into 5 groups at random: Normal group, D-galactose model control group, three tested material groups, often organize 15.Model control group and three tested material groups are all through lumbar injection D-galactose (500mg/kgbw), and injection volume 0.1ml/10gbw, tested material group per os gives the Chinese medicine soft capsule content of the present invention of various dose simultaneously, and matched group gives equivalent Oleum Arachidis hypogaeae semen.D-galactose and the equal successive administration of tested material 60 days, get liver in experiment, measure MDA and SOD after making 10% homogenate latter stage.

1.6.3, MDA assay: measure MDA content in 10% liver homogenate by the requirement of test kit.

1.6.4, SOD vitality test: measure SOD vigor in 10% liver homogenate by the requirement of test kit.

1.7, statistical test is carried out in the variance analysis in experimental data SPSS10.1 for Windows.

2, result:

2.1, Chinese medicinal soft capsule of the present invention is on the impact of life span of drosophila melanogaster:

Table 1 Chinese medicinal soft capsule drosophila survival experiment of the present invention statistical table

Wherein a (Mean longest life) is calculated by 20 fruit bats of longest-lived.

From table 1, in drosophila survival experiment, each group fruit bat vegetative activity is good.The dead natural law of Male Drosophila 0.140% concentration group half extends 6 days than matched group, and average life and Mean longest life all extend than matched group, and difference has significance (P < 0.05); Female Drosophila 0.140% concentration group Mean longest life extends than matched group, and difference has significance (P < 0.05).

2.2, Chinese medicinal soft capsule of the present invention is on the impact of Mouse Weight:

Table 2 is on the impact of D-galactose modeling experiment mice body weight

From table 2, after per os gives the Chinese medicinal soft capsule content of the present invention of various dose, the growth of each treated animal, movable normal, each treated animal weight gain compared with model control group, no significant difference (P > 0.05).

2.3, Chinese medicinal soft capsule of the present invention is on the impact of mice MDA, SOD:

Table 3 is on the impact of mice MDA, SOD

From table 3, after per os gives the Chinese medicinal soft capsule content of the present invention of various dose, in each dosage group liver homogenate, MDA content is all lower than model control group, and difference all has significance (P < 0.05); SOD is all higher than model control group, and wherein high dose group is compared with model control group, and difference has significance (P < 0.05).

3, brief summary:

In drosophila survival experiment, each group fruit bat vegetative activity is good.The dead natural law of Male Drosophila 0.140% concentration group half extends 6 days than matched group, and average life and Mean longest life all extend than matched group, and difference has significance (P < 0.05); Female Drosophila 0.140% concentration group Mean longest life extends than matched group, and difference has significance (P < 0.05); In D-galactose modeling zoopery, after per os gives the Chinese medicinal soft capsule content of the present invention of various dose, the growth of each treated animal, movable normal, each treated animal weight gain compared with model control group, no significant difference (P > 0.05); In the impact of mice MDA, SOD is tested, after per os gives the Chinese medicinal soft capsule content of the present invention of various dose, in each dosage group liver homogenate, MDA content is all lower than model control group, and difference all has significance (P < 0.05); SOD is all higher than model control group, and wherein high dose group is compared with model control group, and difference has significance (P < 0.05).According to " function of health food assessment process and the method for inspection ", Chinese medicinal soft capsule of the present invention has the effect of slow down aging.

test example two, Chinese medicinal soft capsule antifatigue effect of the present invention:

The inspection item of this test are the zoopery of Chinese medicinal soft capsule antifatigue effect of the present invention.

1, materials and methods:

1.1, sample: Chinese medicinal soft capsule of the present invention, content is oily thick liquid, and intake is 1.40/60kgbw day.

1.2, laboratory animal: 18-22g, male secondary Kunming mouse 160, is provided by Jun Ke institute Experimental Animal Center, approval number position SCXK-(army) 2002-001 043820.

1.3, laboratory animal room: SPF level laboratory animal room, the quality certification number: dynamic No. 015th, the word of doctor.Temperature: 20-23 DEG C, humidity: 50-60%.Laboratory animal feedstuff is provided by Tianjin Huarong Animal Science company limited.

1.4, dosage choice: Chinese medicinal soft capsule of the present invention, press human body daily intaking amount 1.40/60kgbw, this EXPERIMENTAL DESIGN matched group and basic, normal, high three dosage groups (often organizing 10), dosage is respectively 0.117g/kg, 0.233g/kg, 0.467g/kg.Namely 5 times of human body recommended amounts, 10 times, 20 times are equivalent to.Sample Oleum Arachidis hypogaeae semen is prepared, and by 0.1ml/10gbw gavage, matched group gives equivalent Oleum Arachidis hypogaeae semen, and each group per os gavage, once a day, measures indices after giving 30 days continuously.

1.5, key instrument and reagent:

Swimming trunk (possessing temperature regulation system); Ultraviolet-visible photometer (UV1221); Automatic clinical chemistry analyzer (Vitalab Seletra2); Blood urea nitrogen test kit (production of Beijing Zhong Sheng biological reagent company); Blood lactase acid glucose sensor (YSI2300).

1.6, experimental technique:

1.6.1, by Chinese medicinal soft capsule content Oleum Arachidis hypogaeae semen of the present invention be mixed with suspension, per os gavage gives tested material 30 days, and gavage volume is 0.1ml/10gbw.

1.6.2, Loaned swimming test: mice continuous per os gavage tested material 30 days, swimming test is carried out in follow procedure regulation heavy burden 2%, and record mice enters swimming time when water to power exhausts dead.

1.6.3, serum urea nitrogen determination: mice continuous per os gavage tested material is after 30 days, and the swimming of follow procedure regulation was taken a blood sample after 90 minutes, and separation of serum also carries out urea nitrogen content mensuration.

1.6.4, hepatic glycogen measures: mice continuous per os gavage tested material 30 days, follow procedure regulation directly measures.

1.6.5, blood lactase acid measures: mice continuous per os gavage tested material is got tail blood and is about 40ul after 30 days, the test tube of 8ulEDTA has been put in instillation, abundant mixing, and then not swimming with a load attached to the body is taken a blood sample after 10 minutes immediately in the water of 30 DEG C, have a rest and take a blood sample again after 20 minutes, blood sampling volume and processing method three times identical.Then measure with lactic acid instrument.According to following formulae discovery blood lactase acid area under curve:

Blood lactase acid area under curve=1/2 × (the blood lactase acid value of the rear 0min of front blood lactase acid value+swimming that swims) × 10+1/2 × (the blood lactase acid value of the rear rest 20min of blood lactase acid value+swimming of the rear 0min of swimming) × 20=5 × (the blood lactase acid value of blood lactase acid value+2 × rear rest 20min that swims of the rear 0min of front blood lactase acid value+3 × swimming that swims).

1.6.6, experimental data statistics: carry out variance analysis and t inspection with SPSS10.0 and Microsoft Excel Software.

2, result:

2.1, Chinese medicinal soft capsule of the present invention is on the impact of Mouse Weight:

Table 4 resisting fatigue one group of Mouse Weight

Table 5 resisting fatigue two groups of Mouse Weights

Table 6 resisting fatigue three groups of Mouse Weights

Table 7 resisting fatigue four groups of Mouse Weights

By table 4-7 visible each dosage group Mouse Weight and matched group more in test phase, latter stage and increase weight that there are no significant for difference.

2.2, Chinese medicinal soft capsule of the present invention is on the impact of mice burden swimming time:

Table 8 is on the impact of mice burden swimming time

From table 8, in Loaned swimming test, each dosage treated animal swimming time is all longer than matched group, and wherein low, high dose group is compared with matched group, and difference has significance (P < 0.05).

2.3, Chinese medicinal soft capsule of the present invention is on the impact of mouse movement serum urea nitrogen after 90 minutes:

Table 9 is on the impact of mouse movement serum urea nitrogen after 90 minutes

From table 9, in serum urea nitrogen test, each dosage group serum urea nitrogen is all lower than matched group, and difference all has significance (P < 0.05).

2.4, Chinese medicinal soft capsule of the present invention is on the impact of Mouse Liver glycogen content:

Table 10 is on the impact of Mouse Liver glycogen content

From table 10, in hepatic glycogen test, each dosage group Mouse Liver glycogen content is compared with matched group, and there are no significant for difference (P > 0.05).

2.5, Chinese medicinal soft capsule of the present invention is on the impact of Mouse Blood lactate level:

Table 11 is on the impact of Mouse Blood lactate level

From table 11, each dosage group Mouse Blood lactic acid area under curve is all lower than matched group, and wherein high dose group is compared with matched group, and difference has significance (P < 0.05).

3, brief summary:

Per os gave the Chinese medicinal soft capsule content of the present invention of various dose after 30 days, in Loaned swimming test, each dosage treated animal swimming time is all longer than matched group, wherein low, high dose group is compared with matched group, and difference has significance (P < 0.05); In serum urea nitrogen test, each dosage group serum urea nitrogen is all lower than matched group, and difference all has significance (P < 0.05); In hepatic glycogen test, each dosage group Mouse Liver glycogen content is compared with matched group, and there are no significant for difference (P > 0.05); During blood lactase acid measures, each dosage group Mouse Blood lactic acid area under curve is all lower than matched group, wherein high dose group is compared with matched group, difference has significance (P < 0.05), has the effect of resisting fatigue by the criterion Chinese medicinal soft capsule of the present invention of " function of health food assessment process and the method for inspection ".

The above, be only preferred embodiment of the present invention, not any formal and substantial restriction is done to the present invention, all those skilled in the art, do not departing within the scope of technical solution of the present invention, when utilizing disclosed above technology contents, and a little change made, modify with differentiation equivalent variations, be Equivalent embodiments of the present invention; Meanwhile, all according to substantial technological of the present invention to the change of any equivalent variations that above embodiment is done, modify and differentiation, all still belong in the scope of technical scheme of the present invention.

Claims (6)

1. a Chinese medicine composition for slow down aging and resisting fatigue, described Chinese medicine composition is made up of the crude drug of following weight portion: Radix Ginseng 2 parts, Semen Vitis viniferae 10 parts, Fructus Tribuli 1 part and Herba Epimedii 1 part.

2. the Chinese medicinal soft capsule of a slow down aging and resisting fatigue, described Chinese medicinal soft capsule comprises active component and pharmaceutically acceptable carrier, it is characterized in that, described active component is prepared by following method by the Radix Ginseng of weight portion described in claim 1, Semen Vitis viniferae, Fructus Tribuli and Herba Epimedii:

(1), insert in extraction pot by Radix Ginseng decoction pieces, with 75% ethanol extraction twice, add ethanol 6 times amount at every turn, extract 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and when decompression recycling ethanol to 60 is spent, the concentrated solution of relative density 1.15 is for subsequent use; Residue stays and does water extraction use;

(2) after, getting the middle Radix Ginseng residue mixing of Semen Vitis viniferae, Fructus Tribuli coarse grain, Herba Epimedii crin and step (1), extracting in water 3 times, add water 6 times amount at every turn, carry 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and leave standstill 4 hours, Aspirate supernatant, the extraction concentrated solution of relative density 1.15 when evaporation and concentration to 60 is spent; This extraction concentrated solution is added the ethanol of 95%, make alcohol content be 60%, leave standstill 24 hours, Aspirate supernatant postprecipitation filters, and filtrate and supernatant merge, decompression recycling ethanol, to the concentrated solution of relative density 1.15 when 60 degree;

(3) after the concentrated solution that the concentrated solution, by step (2) obtained and step (1) obtain mixes, the spray-dried dry extract obtaining active component.

3. Chinese medicinal soft capsule according to claim 2, is characterized in that: containing Total saponin Rb in every 100g active component ₁430mg, procyanidin 253mg.

4. the Chinese medicinal soft capsule according to Claims 2 or 3, is characterized in that: described pharmaceutically acceptable carrier is made up of soybean oil, gelatin, glycerol and distilled water.

5. the preparation method of the Chinese medicinal soft capsule of a slow down aging and resisting fatigue, comprise the step being obtained Chinese medicinal soft capsule by active component and pharmaceutically acceptable carrier, it is characterized in that: described preparation method also comprises the preparation process of active component, wherein the preparation process of active component is prepared by following method by the Radix Ginseng of weight portion described in claim 1, Semen Vitis viniferae, Fructus Tribuli and Herba Epimedii:

(1), insert in extraction pot by Radix Ginseng decoction pieces, with 75% ethanol extraction twice, add ethanol 6 times amount at every turn, extract 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and when decompression recycling ethanol to 60 is spent, the concentrated solution of relative density 1.15 is for subsequent use; Residue stays and does water extraction use;

(2) after, getting the middle Radix Ginseng residue mixing of Semen Vitis viniferae, Fructus Tribuli coarse grain, Herba Epimedii crin and step (1), extracting in water 3 times, add water 6 times amount at every turn, carry 2 hours, 100 order stainless steel sifts filter, and filtrate merges, and leave standstill 4 hours, Aspirate supernatant, the extraction concentrated solution of relative density 1.15 when evaporation and concentration to 60 is spent; This extraction concentrated solution is added the ethanol of 95%, make alcohol content be 60%, leave standstill 24 hours, Aspirate supernatant postprecipitation filters, and filtrate and supernatant merge, decompression recycling ethanol, to the concentrated solution of relative density 1.15 when 60 degree;

(3) after the concentrated solution that the concentrated solution, by step (2) obtained and step (1) obtain mixes, the spray-dried dry extract obtaining active component.

6. preparation method according to claim 5, is characterized in that: the step being obtained Chinese medicinal soft capsule by active component and pharmaceutically acceptable carrier comprises:

A (), the dry extract getting the active component preparing gained mix with the soybean oil of pharmaceutically acceptable amount, through colloid mill, grinding evenly, obtains the suspensoid of dry extract oil;

(b), according to gelatin: glycerol: distilled water=1: the weight ratio of 0.6: 1, takes each raw material, is first soaked in water by gelatin 12 hours, after making it fully expand, the glycerol of additional proportion amount, adds distilled water, is heated to 70-80 degree, stirring, melting, mixing, insulation 1-2 hour, leaves standstill, removing floating foam, filter, obtain glue shell feed liquid heat preservation for standby use;

C glue shell feed liquid that (), the suspensoid of dry extract oil step (a) obtained and step (b) obtain is placed in automatic rotary respectively and presses capsule Bigpian, room temperature 23 degree, under the condition of relative humidity 40%, be pressed into soft gelatin capsule, obtain Chinese medicinal soft capsule.